High-affinity agents targeting prostate-specific membrane antigen for internal radiotherapy of prostate cancer

Abstract

High affinity agents targeting prostate-specific membrane antigen for internal radiotherapy of prostate cancer are disclosed. [Selected Figure] Figure 1

Description

[Background technology]

[0001] [CROSS-REFERENCE TO RELATED APPLICATIONS] This application claims the benefit of U.S. Provisional Application No. 62 / 512,515, filed May 30, 2017, which is incorporated herein by reference in its entirety.

[0002] [Federally funded research or development] This invention was made with government support under grants K25CA148901-01A1 and U54CA1346751 awarded by the National Institutes of Health (NIH). The U.S. Government has certain rights in this invention.

[0003] Prostate cancer is the leading cancer in the United States and the second leading cause of cancer deaths in men. Treatment of locally advanced disease remains controversial, with an increasing number of different options becoming available. Novel, high-affinity, radiotherapy agents for prostate cancer have been developed using prostate-specific membrane antigen (PSMA) as a target. PSMA is a marker of androgen-independent disease, which is also expressed in the neovasculature of solid (non-prostate) tumors. Summary of the Invention [Means for solving the problem]

[0004] [summary] In one embodiment, the subject of the present invention is a compound of formula (I): [ka] wherein Z is tetrazole or COQ; Q is H or a protecting group; m is an integer selected from the group consisting of 1, 2, 3, 4, and 5; and R is independently H or -CH-R 1 and;R 1is selected from the group consisting of substituted aryl, substituted pyridine, and unsubstituted isoquinoline; L is a linker selected from the group consisting of C-C alkylene and C-C cycloalkylene and arylene; W is -NR 2 -(C=O)-, -NR 2 -(C=S)-, -(C=O)-NR 2 - and -(C=S)-NR 2 - wherein each occurrence of L and W may be the same or different; R 2 is H or C1-C4 alkyl; n is an integer selected from the group consisting of 1, 2, and 3; Ch is a chelating agent that can contain a metal or a radioactive metal; and pharmaceutically acceptable salts thereof.

[0005] In a specific embodiment of the compound of formula (I), R 1 is the following: [ka] wherein X is independently Br or I; is selected from the group consisting of:

[0006] In a further embodiment of the compound of Formula (I), the chelating agent is: [ka] is selected from the group consisting of:

[0007] In another aspect, the present invention provides a method comprising contacting one or more PSMA-expressing tumors or cells with an effective amount of a compound of formula (I), wherein said compound of formula (I) is: [ka] wherein Z is tetrazole or COQ; Q is H or a protecting group; m is an integer selected from the group consisting of 1, 2, 3, 4, and 5; and R is independently H or -CH-R 1 and;R 1 is substituted aryl, substituted pyridine, and unsubstituted isoquinoline; L is a linker selected from the group consisting of C1-C6 alkylene and C3-C6 cycloalkylene and arylene; W is -NR 2 -(C=O)-, -NR 2 -(C=S)-, -(C=O)-NR 2 - and -(C=S)-NR 2 - wherein each occurrence of L and W may be the same or different; R 2 is H or C1-C4 alkyl; n is an integer selected from the group consisting of 1, 2, and 3; Ch is a radiometal-containing chelating agent suitable for radiotherapy; and pharmaceutically acceptable salts thereof. The present invention provides a method for treating tumors or cells that express one or more PSMAs, comprising:

[0008] In another aspect, the present subject matter provides a method of imaging one or more prostate-specific membrane antigen (PSMA) tumors or cells, the method comprising contacting the one or more tumors or cells with an effective amount of a compound of formula (I) and imaging.

[0009] In yet another aspect, the present subject matter provides kits comprising compounds of formula (I).

[0010] Certain aspects of the inventive subject matter set forth hereinabove will be described in whole or in part by the inventive subject matter, and other aspects will become apparent as best described hereinafter when read in conjunction with the accompanying examples and drawings. [Brief explanation of the drawings]

[0011] Having described the subject matter of the present invention in general terms, reference is made to the accompanying drawings, which are not necessarily drawn to scale, in which: [Figure 1] Figure 1 shows the chemical structures of representative radiotherapy agents; [Figure 2] Figure 2 shows a comparative study of the efficacy of 177Lu-8 and known drugs SR6, PSMA-617 and PSMA-I&T in clonogenesis; [Figure 3] Figure 3 shows the distribution ratio of 177Lu-8, 177Lu-1, 177Lu-SR6, 177Lu-PSMA-617, and 177Lu-PSMA-I&T to PSMA+ tumors relative to the kidney; [Figure 4] Figure 4 shows SPECT-CT imaging of 177Lu-8 during a treatment study using a single dose of 3 mCi; [Figure 5] Figure 5 shows the relative body weight of mice during the treatment study; [Figure 6] Figures 6A and 6B show the relative tumor volumes of mice during the treatment study ( Figure 6A ) and Kaplan-Meier survival curves up to 60 days after treatment ( Figure 6B ); [Figure 7] Figure 7 shows (left) in vitro cellular uptake of 213Bi-1 at 37°C. Specific cellular uptake in PSMA(+)PC3 PIP cells (blue) and PSMA(-)PC3 flu cells (black), as well as when blocked with 1 mM ZJ43 (red). (middle and right) Specific cell killing in response to 213Bi-1 and 225 Ac-1 after 2 hours at 37°C. [Figure 8] Figure 8 shows the tissue biodistribution of 213Bi-1 (% ID / g); [Figure 9]Figure 9 shows the efficacy of 225Ac-1 in the PSMA+ PIP flank tumor model (male NSG mice) (top). (middle) Changes in body weight during the treatment study. (bottom) Kaplan-Meier survival curves for the treatment study using the micrometastatic PC3-ML-Luc-PSMA model, with a median survival of 51 days (untreated) vs. 52 days for 177Lu-PSMA-617 (37 MBq), 56 days for 225Ac-1 (37 kBq), and 79 days for 225Ac-1 (74 kBq); [Figure 10] Figure 10 shows SPECT-CT imaging (top) and tissue biodistribution data (bottom) of 203Pb-2 and 203Pb-3 (14.8 MBq.). T+: PSMA(+)PC3 PIP, T(-) PC3 flu; [Figure 11] Figures 11A, 11B, 11C, 11D, and 11E show the distribution of 0.37 MBq 225Ac-1 (A) and 1.85 MBq 213Bi-1 (+2 nmol 9) (B) in PSMA+ PC3 PIP tumors (T) and kidneys (K), as recorded by alpha camera imaging. Co-injection with 2 nmol 9 specifically cleared activity from the renal cortex but not from PSMA+ tumors. (C) IHC of 225Ac-9-treated PSMA+ tumors shows decreased PSMA-expression after 24 hours compared to untreated PSMA+ tumors. (D, E) PSMA+ staining (IHC) of kidneys from the same mice as in (A) after 2 hours; [Figure 12] Figure 12 shows the biodistribution of 177Lu-1 with increasing amounts of 1, demonstrating that it is a partial renal blocking ligand at 2 hours; [Figure 13] Figures 13A and 13B show the first patient with extensive metastatic disease scanned with type I (high tumor and kidney uptake) and type II (high tumor and low kidney uptake) contrast agents at 2 hours post-injection, and with 68Ga-2 (Figure 13A) and 68Ga-2 (Figure 13B). Subcentimeter perihilar and thoracic lymph nodes were included (axial panel, arrows) (left) (Reference 34); selected 68Ga / 177Lu-labeled PSMA-binding agents (right); [Figure 14]Figure 14 shows comparative cellular uptake and internalization data for selected compounds (head-to-head study); [Figure 15] Figure 15 shows comparative tissue biodistribution data for selected compounds (head-to-head study); [Figure 16] Figure 16 shows comparative tissue biodistribution data for selected compounds (head-to-head studies, also shown in Figure 15 ); [Figure 17] Figure 17 shows the biodistribution data of 177Lu-1, 177Lu-7, 177Lu-8, 177Lu-9, 177Lu-10, and 177Lu-14; [Figure 18] Figures 18A and 18B show SPECT / CT imaging with 177Lu-1 (37 MBq) (Figure 18A); Figure 18B. Immunohistochemistry for PSMA expression in PC3 PIP tumors administered without or with 37 MBq of 177Lu-1; [Figure 19] Figure 19 shows SPECT / CT imaging after intravenous administration of 177Lu-14; [Figure 20] Figure 20 shows the partial autoblockade of 177Lu-1 and 177Lu-9 at 2 hours post-injection; [Figure 21] Figures 21A and 21B show radionuclide therapy with PSMA-based 177Lu-labeled small molecular weight agents 177Lu-1, 177Lu-3, 177Lu-5, and 177Lu-PSMA-617. Tumor growth inhibition (Figure 21A) and survival (Figure 21B) after a single dose of ~111 MBq of each agent via tail vein injection and control saline injection (n = 10 for 177Lu-1, 177Lu-3, 177Lu-5, n = 9 for 177Lu-PSMA-617). Relative tumor volumes and body weights of individual mice are shown after 4 and 8 weeks of treatment; [Figure 22] Figure 22 shows the relative volume, body weight and survival rate (%) of 111 MBq of 177Lu-PSMA-617 and 177Lu-1; [Figure 23]Figure 23 shows hemoglobin and blood counts 8 weeks after radiation therapy with 177Lu-1, 177Lu-3, 177Lu-5, and 177Lu-PSMA-617. Data are mean ± SEM, n = 3 / group; [Figure 24] Figure 24 shows 177Lu-1 radionuclide therapy after a single dose of 111 MBq administered via tail vein injection, followed by a saline control injection (n = 10 for 177Lu-1, 177Lu-3, 177Lu-5, n = 9 for 177Lu-PSMA-617). Relative tumor volumes and body weights of individual mice are shown after 4 and 8 weeks of treatment; and [Figure 25] Figures 25A, 25B, 25C, 25D, and 25E show hematoxylin and eosin (H&E) staining. (Figures 25A, 25B, and 25C) Extraorbital lacrimal gland: Figure 25A. Control untreated; Figure 25B. Adjacent parotid gland (arrowhead); Figure 25C. Higher magnification of Figure 25B (177Lu-PSMA-617) showing acinar loss with chronic and active inflammation affecting the lacrimal gland (Figures 25D and 25E). Testis: Figure 25D. Control untreated, active spermatogenesis; some tubules (arrowheads) contain only Sertoli cells, as expected near the rete testis; Figure 25E. Spermatogenesis is absent, and the diffuse seminiferous tubules are smaller than in A (same magnification), lined only with Sertoli cells, with a few germ cells in some tubules. In this specimen, interstitial cells are prominent between the tubules. The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. DETAILED DESCRIPTION OF THE INVENTION

[0012] [Detailed explanation] The present subject matter is described more fully below with reference to the accompanying drawings, in which some, but not all, embodiments of the invention disclosed herein are shown. Like numbers refer to like elements throughout. The present subject matter may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Indeed, many modifications and other embodiments of the invention disclosed herein will come to mind to one skilled in the art to which the present subject matter pertains having the benefit of the teachings presented in the foregoing descriptions and the accompanying drawings. Therefore, it is to be understood that the invention disclosed herein is not to be limited to the specific embodiments disclosed, and that modifications and other embodiments are intended to be included within the scope of the appended claims.

[0013] I. High-affinity drugs targeting prostate-specific membrane antigen for internal radiotherapy of prostate cancer

[0014] A. Compounds of Formula (I)

[0015] Thus, in one embodiment, the subject of the present invention is a compound of formula (I): [ka] wherein Z is tetrazole or COQ; Q is H or a protecting group; m is an integer selected from the group consisting of 1, 2, 3, 4, and 5; and R is independently H or -CH-R 1 and;R 1 is selected from the group consisting of substituted aryl, substituted pyridine, and unsubstituted isoquinoline; L is a linker selected from the group consisting of C-C alkylene and C-C cycloalkylene and arylene; W is -NR 2 -(C=O)-, -NR 2 -(C=S)-, -(C=O)-NR 2 - and -(C=S)-NR 2- wherein each occurrence of L and W may be the same or different; R 2 is H or C1-C4 alkyl; n is an integer selected from the group consisting of 1, 2, and 3; Ch is a chelating agent that can contain a metal or a radioactive metal; and pharmaceutically acceptable salts thereof.

[0016] The phrase "wherein each occurrence of L and W can be the same or different" means that when the variable "n" is 2 or 3, one "L" group can be C1-C6 alkylene, and the other "L" group or groups can be C3-C6 cycloalkylene or arylene, or in other embodiments, each "L" group can be, for example, C1-C6 alkylene. Similarly, when "n" is 2 or 3, for example, one "W" group can be -(C=O)-NR 2 - and the other "W" group or groups may be -(C=S)-NR 2 or in other embodiments, each "W" may be, for example, -(C=O)-NR 2 - It's okay.

[0017] In specific embodiments of the compounds of formula (I), R 1 is the following: [ka] wherein X is independently Br or I; is selected from the group consisting of:

[0018] In a further embodiment of the compound of formula (I), the chelating agent is: [ka] is selected from the group consisting of:

[0019] In more specific embodiments of the compounds of Formula (I), the chelator comprises a metal selected from the group consisting of: Y, Lu, Tc, Zr, In, Sm, Re, Cu, Pb, Ac, Bi, Al, Ga, Re, Ho, and Sc. In further specific embodiments of the compounds of Formula (I), the metal is a radiometal and is selected from the group consisting of: 68 Ga, 64 Cu, 86 Y, 90 Y, 89 Zr, 111 In, 99m Tc, 177 Lu, 153 Sm, 186 Re, 188 Re, 67 Cu, 212 Pb, 225 Ac, 213 Bi, 212 Bi, 212 Pb, 67 Ga, 203 Pb, 47 Sc and 166 Ho.

[0020] In a specific embodiment, the compound of formula (I) is: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] is selected from the group consisting of:

[0021] B. Methods of Using Compounds of Formula (I) to Treat One or More PSMA-Expressing Tumors or Cells

[0022] In one embodiment, the present invention is directed to a method comprising contacting one or more PSMA-expressing tumors or cells with an effective amount of a compound of formula (I), wherein said compound of formula (I) is: [ka] wherein Z is tetrazole or COQ; Q is H or a protecting group; m is an integer selected from the group consisting of 1, 2, 3, 4, and 5; and R is independently H or -CH-R 1 and;R 1 is selected from the group consisting of substituted aryl, substituted pyridine, and unsubstituted isoquinoline; L is a linker selected from the group consisting of C-C alkylene and C-C cycloalkylene and arylene; W is -NR 2 -(C=O)-, -NR 2 -(C=S)-, -(C=O)-NR 2 - and -(C=S)-NR 2 - wherein each occurrence of L and W may be the same or different; R 2is H or C1-C4 alkyl; n is an integer selected from the group consisting of 1, 2, and 3; Ch is a radiometal-containing chelating agent suitable for radiotherapy; and pharmaceutically acceptable salts thereof. The present invention provides a method for treating tumors or cells that express one or more PSMAs, comprising:

[0023] "Contacting" refers to any act of bringing at least one compound, including a therapeutic agent of the present invention, into physical contact with at least one tumor or cell expressing PSMA. Contacting may include exposing a cell or tumor to at least one compound in an amount sufficient to contact the at least one cell or tumor. The method may be performed in vitro or ex vivo by placing and preferably mixing the compound and cell or tumor in a controlled environment, such as a culture dish or tube. The method may be performed in vivo, in which case contacting refers to exposing at least one cell or tumor of a subject to at least one compound of the present invention, such as by administering the compound to the subject by any suitable route.

[0024] As used herein, the term "treating" can include reversing, alleviating, inhibiting the progression of, preventing, or reducing the likelihood of the disease, disorder, or condition to which such term applies, or one or more symptoms or manifestations of such disease, disorder, or condition. Prevention means preventing the disease, disorder, or condition, or any such symptom or manifestation, from occurring or from worsening in severity. Thus, the compounds of the invention can be administered prophylactically to prevent or reduce the occurrence or recurrence of the disease, disorder, or condition.

[0025] Generally, an "effective amount" of an active agent refers to the amount necessary to elicit a desired biological response. As will be appreciated by those skilled in the art, the effective amount of an agent or device may vary depending on factors such as the desired biological endpoint, the agent being delivered, the composition of the pharmaceutical composition, and the target tissue.

[0026] The term "combination" is used in its broadest sense and refers to the administration of at least two pharmaceutical agents, more specifically, a compound of Formula (I) and at least one other active agent, to a subject. More specifically, the term "in combination" refers to the administration of two (or more) active agents concomitantly, e.g., for the treatment of a single disease state. As used herein, the active agents may be administered in combination in a single agent, simultaneously as separate agents, or as separate agents administered alternately or sequentially on the same or different days. In some embodiments of the invention disclosed herein, the active agents are administered in combination in a single agent. In other embodiments, the active agents are administered in separate agents (e.g., where it is desirable to vary the amount of one agent while maintaining the amount of the other). The single agent may include additional active agents for treating a disease state.

[0027] In specific embodiments, R 1 is the following: [ka] wherein X is independently Br or I; is selected from the group consisting of:

[0028] In more specific embodiments, the chelating agent is: [ka] is selected from the group consisting of:

[0029] In a more specific embodiment, the radiometal suitable for radiotherapy is: 90 Y, 177 Lu, 211 At, 111 In, 153 Sm, 186 Re, 188 Re, 67 Cu, 212 Pb, 225 Ac, 213 Bi, 212 Bi, 212 Pb and 67 Ga.

[0030] In a more specific embodiment, the compound of formula (I) is: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] is selected from the group consisting of:

[0031] In other embodiments, the one or more PSMA-expressing tumors or cells are selected from the group consisting of prostate tumors or cells, metastatic prostate tumors or cells, lung tumors or cells, kidney tumors or cells, glioblastoma, pancreatic tumors or cells, bladder tumors or cells, sarcoma, melanoma, breast tumors or cells, colon tumors or cells, germ cells, pheochromocytoma, esophageal tumors or cells, gastric tumors or cells, and combinations thereof. In certain other embodiments, the one or more PSMA-expressing tumors or cells are prostate tumors or cells.

[0032] In other embodiments, the one or more PSMA-expressing tumors or cells are in vitro, in vivo, or ex vivo. In yet other embodiments, the one or more PSMA-expressing tumors or cells are present in a subject.

[0033] Although it should be understood that the methods described herein are effective for all vertebrate species intended to be encompassed by the term "subject," in many embodiments, the subject treated by the methods of the invention is desirably a human subject. Thus, a "subject" can include a human subject for medical purposes, such as treating an existing disease or disorder or providing prophylactic treatment to prevent the onset of a disease or disorder, or an animal (non-human) subject for medical, veterinary, or development purposes. Suitable animal subjects include mammals, including, but not limited to, primates (e.g., humans, monkeys, apes, etc.); bovines (e.g., cattle, bulls, etc.); ovines (e.g., sheep, etc.); caprines (e.g., goats, etc.); porcines (e.g., pigs, hogs, etc.); equines (e.g., horses, donkeys, zebras, etc.); felines (including wild and pet cats); canines (including dogs); lagomorphs (including rabbits, hares, etc.); rodents (including mice, rats, etc.); and the like. The animal may be a transgenic animal. In one embodiment, the subject is a human, including, but not limited to, fetal, neonatal, infant, juvenile, and adult subjects. Additionally, a "subject" may include a patient suffering from or suspected of suffering from a disease or disorder. Thus, the terms "subject" and "patient" are used interchangeably herein.

[0034] In yet certain other embodiments, the method results in an inhibition of growth of the tumor.

[0035] C. Methods of Using Compounds of Formula (I) to Image Tumors or Cells Expressing One or More PSMAs

[0036] In another embodiment, the present invention is directed to a method comprising contacting one or more PSMA-expressing tumors or cells with an effective amount of a compound of formula (I) and imaging, wherein said compound of formula (I) is: [ka] wherein Z is tetrazole or COQ; Q is H or a protecting group; m is an integer selected from the group consisting of 1, 2, 3, 4, and 5; and R is independently H or -CH-R 1 and;R 1 is selected from the group consisting of substituted aryl, substituted pyridine, and unsubstituted isoquinoline; L is a linker selected from the group consisting of C-C alkylene and C-C cycloalkylene and arylene; W is -NR 2 -(C=O)-, -NR 2 -(C=S)-, -(C=O)-NR 2 - and -(C=S)-NR 2 - wherein each occurrence of L and W may be the same or different; R 2 is H or C1-C4 alkyl; n is an integer selected from the group consisting of 1, 2, and 3; Ch is a chelator containing a radiometal suitable for imaging; and pharmaceutically acceptable salts thereof. The present invention provides a method for imaging tumors or cells that express one or more PSMAs, comprising:

[0037] D. Kit

[0038] In yet another embodiment, the present subject matter provides kits comprising a compound of formula (I).

[0039] In some embodiments, the kit provides a packaged pharmaceutical composition comprising a pharmaceutically acceptable carrier and a compound of the invention. In some embodiments, the packaged pharmaceutical composition comprises the reaction precursors necessary to produce a compound of the invention in combination with a radiolabeled precursor. Other packaged pharmaceutical compositions provided by the invention further comprise instructions comprising at least one of the following: instructions for preparing a compound of the invention from the provided precursors, instructions for using the composition to image cells or tissues expressing PSMA, instructions for using the compound to image glutamatergic neurotransmission in patients suffering from stress-related disorders, or instructions for using the composition to image prostate cancer.

[0040] E. Pharmaceutical Compositions and Administration

[0041] In another aspect, the present invention provides pharmaceutical compositions comprising a compound of Formula (I), alone or in combination with one or more additional therapeutic agents in admixture with a pharmaceutically acceptable excipient. Those skilled in the art will recognize that the pharmaceutical compositions include pharmaceutically acceptable salts of the compounds described above. Pharmaceutically acceptable salts are widely known to those skilled in the art and include salts of active compounds prepared with relatively non-toxic acids or bases, depending on the particular substituent moieties found on the compounds described herein. When a compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such a compound, neat or in a suitable inert solvent, with a sufficient amount of the desired base, or by ion exchange (substituting another for the basic counterion (base) in an ionic complex). Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts.

[0042] When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such a compound, neat or in a suitable inert solvent, with a sufficient amount of the desired acid, or by ion exchange (substituting one acidic counterion (acid) in an ionic complex for another). Examples of pharmaceutically acceptable acid addition salts include salts 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, as well as salts derived from relatively non-toxic 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, and the like. Also included are salts of amino acids, such as arginate, and salts of organic acids, such as glucuronic acid or galacturonic acid (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 functionalities that allow the compounds to be converted into either base or acid addition salts.

[0043] Thus, pharmaceutically acceptable salts suitable for use with the subject matter of the present invention include, by way of non-limiting example, acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, carnsylate, carbonate, citrate, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, , hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, mucate, napsylate, nitrate, pamoate (embonate), pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, diacetate, succinate, sulfate, tannate, tartrate, or teoclate. Other pharmaceutically acceptable salts are described, for example, in Remington: The Science and Practice of Pharmacy (20th ed.), Lippincott, Williams & Wilkins (2000).

[0044] In therapeutic and / or diagnostic applications, the compounds of the invention can be formulated for a variety of modes of administration, including systemic, topical, or localized administration. Techniques and formulations are generally described in Remington: The Science and Practice of Pharmacy (20th ed.), Lippincott, Williams & Wilkins (2000).

[0045] Depending on the specific condition being treated, such agents can be formulated into liquid or solid dosage forms and administered systemically or locally. The agents can be delivered, for example, in sustained-release or sustained-release forms, as known to those skilled in the art. Techniques for formulation and administration are described in Remington: The Science and Practice of Pharmacy (20th ed.), Lippincott, Williams & Wilkins (2000). Suitable routes may include oral, buccal, inhalation spray, sublingual, rectal, transdermal, vaginal, transmucosal, nasal, or intestinal administration; intramuscular, subcutaneous, or intramedullary injection; and parenteral or other delivery modes, including intrathecal, direct intraventricular, intravenous, intraarticular, intrasternal, intrasynovial, intrahepatic, intralesional, intracranial, intraperitoneal, intranasal, or intraocular injection.

[0046] For injection, the agents of the present invention can be formulated and diluted in aqueous solutions, such as Hank's solution, Ringer's solution, or physiologically compatible buffers such as physiological saline buffer. For such transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0047] To practice the present invention, the use of pharmaceutically acceptable inert carriers to formulate the compounds disclosed herein into formulations suitable for systemic administration is within the scope of the present invention. By appropriately selecting carriers and manufacturing procedures, the compositions of the present invention, particularly those formulated as solutions, can be administered parenterally, such as by intravenous injection. The compounds can be easily formulated into formulations suitable for oral administration using pharmaceutically acceptable carriers well known in the art. Such carriers allow the compounds of the present invention to be formulated as tablets, pills, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by a subject (e.g., a patient) to be treated.

[0048] For nasal or inhalation delivery, the agents of the present invention may also be formulated by methods known to those skilled in the art and may include, for example, but not limited to, solubilizers, diluents, or dispersing agents such as saline; preservatives such as benzyl alcohol; absorption enhancers; and fluorocarbons.

[0049] Pharmaceutical compositions suitable for use in the present invention include compositions containing the active ingredient in an amount effective to achieve its intended purpose. Determining such an effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. In general, the compounds of the present invention are effective over a wide dosage range. For example, in the treatment of adult humans, dosages of 0.01 to 1000 mg per day, 0.5 to 100 mg, 1 to 50 mg, and 5 to 40 mg per day are exemplary dosages that may be used. A non-limiting dosage is 10 to 30 mg per day. The exact dosage will depend on the route of administration, the form in which the compound is administered, the subject being treated, the subject's body weight, the bioavailability of the compound, the absorption, distribution, metabolism, and excretion (ADME) profile of the compound, toxicity, and the preference and experience of the attending physician.

[0050] In addition to the active ingredient, these pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers, including excipients and auxiliaries that facilitate processing of the active compound into pharmaceutically usable preparations. Preparations formulated for oral administration may take the form of tablets, dragees, capsules or solutions.

[0051] Oral pharmaceutical preparations can be obtained by mixing the active compound with solid excipients, optionally with the addition of suitable auxiliaries, to obtain tablets or dragee cores, optionally milling the resulting mixture, and processing the resulting granules. Suitable excipients include, inter alia, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose (CMC), and / or polyvinylpyrrolidone (PVP: povidone). Disintegrants such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof (e.g., sodium alginate) can also be added, if desired.

[0052] Dragee cores are provided with a suitable coating. For this purpose, concentrated sugar solutions, optionally containing gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol (PEG), and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures, can be used. Dye stuffs or pigments may be added to the tablets or dragee coatings to identify or characterize different combinations of active compound doses.

[0053] Orally available pharmaceutical formulations include push-fit capsules made of gelatin and soft, sealed capsules made of gelatin and containing a plasticizer such as glycerol or sorbitol. Push-fit capsules may contain the active ingredient in admixture with fillers such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, the active compound can be dissolved or suspended in a suitable liquid, such as fatty oils, liquid paraffin, or liquid polyethylene glycol (PEG). Additionally, stabilizers may be added.

[0054] II. General Definitions

[0055] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter of this invention belongs.

[0056] While the following terms relating to compounds of formula (I) are believed to be well understood by those of ordinary skill in the art, the following definitions are provided to facilitate discussion of the subject matter of the present invention. These definitions are intended to supplement and explain, but not preclude, definitions that would be apparent to those of ordinary skill in the art upon consideration of the present invention.

[0057] As used herein, the terms substituted and substitute, whether preceded by the term "optionally," mean that one functional group on a molecule can be changed to another functional group, provided that the valences of all atoms are maintained, as would be understood by one of ordinary skill in the art. When more than one position in any given structure can be substituted with more than one substituent selected from a specified group, the substituents can be the same or different at all positions. The substituents can also be further substituted (e.g., an aryl group substituent can be replaced with another substituent, such as another aryl group, which is further substituted at one or more positions).

[0058] Where substituents or linking groups are specified by their conventional chemical formula written from left to right, they equally encompass the chemically identical substituents that would result by writing the structure from right to left (e.g., -CHO- is equivalent to -OCH-; -C(=O)O- is equivalent to -OC(=O)-; -OC(=O)NR- is equivalent to -NRC(=O)O-, etc.).

[0059] When the term "independently selected" is used, the substitutions referred to (e.g., R groups such as groups R1, R2, or variables such as "m" and "n") can be the same or different. For example, both R1 and R2 can be substituted alkyl, or R1 can be hydrogen and R2 can be substituted alkyl, etc.

[0060] The terms "a," "an," or "a(n)," when used in reference to a group of substituents herein, mean at least one. For example, if a compound is substituted with "an" alkyl or aryl, then said compound is optionally substituted with at least one alkyl and / or at least one aryl. Furthermore, when a moiety is substituted with an R substituent, the group may be referred to as "R-substituted." When a moiety is R-substituted, said moiety is substituted with at least one R substituent, and each R substituent is optionally different.

[0061] A named "R" or group generally has the structure recognized in the art as corresponding to the group having that name, unless otherwise specified herein. For purposes of illustration, certain representative "R" groups listed above are defined below.

[0062] The description of the compounds of the present invention is limited by principles of chemical bonding known to those skilled in the art. Thus, when a group may be substituted with one or more substituents, such substituents are selected in accordance with principles of chemical bonding and are not inherently unstable and / or may be unstable under ambient conditions, such as aqueous, neutral, and some known physiological conditions, as recognized by those skilled in the art. For example, a heterocycloalkyl or heteroaryl is bonded to the remainder of the molecule via a ring heteroatom in accordance with principles of chemical bonding known to those skilled in the art, thereby avoiding inherently unstable compounds.

[0063] Unless expressly defined otherwise, a "substituent group," as used herein, includes a functional group selected from one or more of the following moieties, as defined herein:

[0064] The term hydrocarbon, as used herein, refers to any chemical group containing hydrogen and carbon. The hydrocarbon may be substituted or unsubstituted. As one skilled in the art would know, all valences must be satisfied with any substitutions. The hydrocarbon may be unsaturated, saturated, branched, unbranched, cyclic, polycyclic, or heterocyclic. Exemplary hydrocarbons are further defined herein below and include, for example, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, allyl, vinyl, n-butyl, tert-butyl, ethynyl, cyclohexyl, and the like.

[0065] The term “alkyl,” by itself or as part of another substituent, means, unless otherwise stated, a straight-chain (i.e., “unbranched”), branched-chain, acyclic, or cyclic hydrocarbon group, or combinations thereof, which may be fully saturated, mono- or polyunsaturated, and which may include divalent and polyvalent groups, having the specified number of carbon atoms (i.e., C1-C6). 10 means 1 to 10 carbons, including 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 carbons. In specific embodiments, the term "alkyl" refers to groups containing 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 carbons. 1-20 means, inclusive, linear (i.e., "straight-chain"), branched, or cyclic, saturated or at least partially, and in some cases fully unsaturated (i.e., alkenyl and alkynyl) hydrocarbon radicals derived by removing one hydrogen atom from a hydrocarbon moiety containing from 1 to 20 carbon atoms.

[0066] Representative saturated hydrocarbon groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, isopentyl, neopentyl, n-hexyl, sec-hexyl, n-heptyl, n-octyl, n-decyl, n-undecyl, dodecyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, and homologs and isomers thereof.

[0067] "Branched" refers to an alkyl group in which a lower alkyl group, such as methyl, ethyl, or propyl, is attached to a linear alkyl chain. "Lower alkyl" refers to an alkyl group having 1 to about 8 carbon atoms, e.g., 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms (i.e., C 1-8 "Higher alkyl" refers to an alkyl group having from about 10 to about 20 carbon atoms, e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. In certain embodiments, "alkyl" refers to, specifically, C 1-8 In another embodiment, "alkyl" refers to, in particular, C 1-8 Refers to branched chain alkyl.

[0068] An alkyl group can be optionally substituted with one or more alkyl group substituents ("substituted alkyl"), which can be the same or different. The term "alkyl group substituent" includes, but is not limited to, alkyl, substituted alkyl, halo, arylamino, acyl, hydroxyl, aryloxyl, alkoxyl, alkylthio, arylthio, aralkyloxyl, aralkylthio, carboxyl, alkoxycarbonyl, oxo, and cycloalkyl. Optionally, there can be inserted along the alkyl chain one or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms, where the nitrogen substituent is hydrogen, lower alkyl (also referred to herein as "alkylaminoalkyl"), or aryl.

[0069] Thus, as used herein, the term "substituted alkyl" includes alkyl groups, as defined herein, in which one or more atoms or functional groups of the alkyl group have been replaced with another atom or functional group (including, for example, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino, dialkylamino, sulfate, mercapto, and the like).

[0070] The term "heteroalkyl," alone or in combination with other terms, unless otherwise stated, means a stable linear, branched, or cyclic hydrocarbon radical, or combination thereof, consisting of at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, Si, and S, wherein the nitrogen, phosphorus, and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) O, N, P, S, and Si may be located at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. Non-limiting examples include -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH 25 These include -S(O)-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, O-CH3, -O-CH2-CH3 and -CN. Up to two or three heteroatoms may be consecutive, e.g., -CH2-NH-OCH3 and -CH2-O-Si(CH3)3.

[0071] As noted above, as used herein, heteroalkyl groups include groups attached to the remainder of the molecule via a heteroatom, such as -C(O)NR', ​​-NR'R", -OR', -SR, -S(O)R, and / or -S(O)R'. When "heteroalkyl" is recited followed by a list of specific heteroalkyl groups, such as -NR'R, it is understood that the terms heteroalkyl and -NR'R" are not overlapping or mutually exclusive. Rather, the specific heteroalkyl groups are listed for clarity. Thus, the term "heteroalkyl" should not be construed herein as excluding specific heteroalkyl groups, such as -NR'R".

[0072] "Cyclic" and "cycloalkyl" refer to a non-aromatic monocyclic or polycyclic ring system of about 3 to about 10 carbon atoms (e.g., 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms). The cycloalkyl group may optionally be partially unsaturated. The cycloalkyl group may also be optionally substituted with alkyl group substituents, oxo, and / or alkylene, as defined herein. Optionally inserted along the cyclic alkyl chain is one or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms, where the nitrogen substituent is hydrogen, unsubstituted alkyl, substituted alkyl, aryl, or substituted aryl, thus providing a heterocyclic group. Representative monocyclic cycloalkyl rings include cyclopentyl, cyclohexyl, and cycloheptyl. Multicyclic cycloalkyl rings include adamantyl, octahydronaphthyl, decalin, camphor, camphane, and noradamantyl, as well as fused ring systems such as, for example, dihydro- and tetrahydronaphthalene.

[0073] The term "cycloalkylalkyl," as used herein, refers to a cycloalkyl group, as defined above, attached to the parent molecular moiety through an alkyl group, also defined above. Examples of cycloalkylalkyl groups include cyclopropylmethyl and cyclopentylethyl.

[0074] The terms "cycloheteroalkyl" or "heterocycloalkyl" refer to a non-aromatic, unsaturated, or partially unsaturated ring system, such as a 3- to 10-membered substituted or unsubstituted cycloalkyl ring system containing one or more heteroatoms, which may be the same or different, selected from the group consisting of nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), and silicon (Si), and which may optionally contain one or more double bonds.

[0075] Cycloheteroalkyl rings may be optionally fused to or attached to other cycloheteroalkyl rings and / or non-aromatic hydrocarbon rings. Heterocycles include heterocycles having 1 to 3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. In certain embodiments, the term heterocyclic refers to a non-aromatic 5-, 6-, or 7-membered ring or polycyclic group in which at least one ring atom is a heteroatom selected from O, S, and N (wherein the nitrogen and sulfur heteroatoms can be optionally oxidized), including, but not limited to, bicyclic or tricyclic groups, including fused 6-membered rings having 1 to 3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, where (i) each 5-membered ring has 0 to 2 double bonds, each 6-membered ring has 0 to 2 double bonds, and each 7-membered ring has 0 to 3 double bonds, (ii) the nitrogen and sulfur heteroatoms can be optionally oxidized, (iii) the nitrogen heteroatom can be optionally quaternized, and (iv) any of the above heterocycles can be fused to an aryl or heteroaryl ring. Representative cycloheteroalkyl ring systems include, but are not limited to, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperidyl, piperazinyl, indolinyl, quinuclidinyl, morpholinyl, thiomorpholinyl, thiadiazinanyl, tetrahydrofuranyl, and the like.

[0076] The terms "cycloalkyl" and "heterocycloalkyl," alone or in combination with other terms, represent, unless otherwise stated, cyclic versions of "alkyl" and "heteroalkyl," respectively. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Non-limiting examples of cycloalkyl include cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Non-limiting examples of heterocycloalkyl include 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. The terms "cycloalkylene" and "heterocycloalkylene" refer to the divalent derivatives of cycloalkyl and heterocycloalkyl, respectively.

[0077] Unsaturated alkyl groups are alkyl groups having one or more double or triple bonds. Non-limiting examples of unsaturated alkyl groups include vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and isomers. Alkyl groups limited to hydrocarbon groups are referred to as "homoalkyl."

[0078] More specifically, the term "alkenyl" as used herein encompasses a straight-chain or branched hydrocarbon moiety having at least one carbon-carbon double bond formed by the removal of a single hydrogen molecule, such as C 1-20Alkenyl groups include, for example, ethenyl (i.e., vinyl), propenyl, butenyl, 1-methyl-2-buten-1-yl, pentenyl, hexenyl, octenyl, allenyl, and butadienyl.

[0079] The term "cycloalkenyl" as used herein refers to a cyclic hydrocarbon containing at least one carbon-carbon double bond. Examples of cycloalkenyl groups include cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadiene, cyclohexenyl, 1,3-cyclohexadiene, cycloheptenyl, cycloheptatrienyl, and cyclooctenyl.

[0080] As used herein, the term "alkynyl" refers to a straight or branched chain C alkyl group of the designated number of carbon atoms containing at least one carbon-carbon triple bond. 1-20 It refers to a monovalent group derived from a hydrocarbon. Examples of "alkynyl" include ethynyl, 2-propynyl (propargyl), 1-propynyl, pentynyl, hexynyl, and heptynyl groups.

[0081] The term "alkylene," alone or as part of another substituent, refers to a straight- or branched-chain divalent aliphatic hydrocarbon group derived from an alkyl group having 1 to about 20 carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms). The alkylene group may be straight-chain, branched, or cyclic. The alkylene group may also be optionally unsaturated and / or substituted with one or more "alkyl group substituents." One or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms (also referred to herein as "alkylaminoalkyl") can be optionally inserted along the alkylene group, where the nitrogen substituent is alkyl, as previously described. Exemplary alkylene groups include methylene (-CH2-); ethylene (-CH2-CH2-); propylene (-(CH2)3-); cyclohexylene (-CH6-); 10 -);-CH=CH-CH=CH-;-CH=CH-CH2-;-CH2CH2CH2CH2-, -CH2CH=CHCH2-, -CH2CsCCH2-, -CH2CH2CH(CH2CH2CH3)CH2-, -(CH2) q -N(R)-(CH2) r-, where each of q and r is independently an integer from 0 to about 20 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20), and R is hydrogen or lower alkyl; methylenedioxyl (-O-CH-O-); and ethylenedioxyl (-O-(CH)-O-). Alkylene groups can have from about 2 to about 3 carbon atoms, and even more often have from 6 to 20 carbon atoms. Typically, alkyl (or alkylene) groups have from 1 to 24 carbon atoms, with groups having 10 or fewer carbon atoms being some embodiments of the present invention. A "lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, generally having eight or fewer carbon atoms.

[0082] The term "heteroalkylene," alone or as part of another substituent, means a divalent radical derived from heteroalkyl (including, but not limited to, -CH-CH-S-CH-CH- and -CH-S-CH-CH-NH-CH-). For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxo, alkylenedioxo, alkyleneamino, alkylenediamino, and the like). Furthermore, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied by the direction in which the formula of the linking group is written. For example, the formula -C(O)OR'- represents both -C(O)OR'- and -R'OC(O)-.

[0083] The term "aryl," unless otherwise specified, means an aromatic hydrocarbon substituent which may be monocyclic or polycyclic (e.g., 1 to 3 rings) that are fused or covalently linked together. The term "heteroaryl" refers to an aryl group (or ring) containing 1 to 4 heteroatoms (in the case of multiple rings, each in a separate ring) selected from N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom is optionally quaternized. A heteroaryl group may be attached to the remainder of the molecule through a carbon or heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isooxazolyl, and 5-isooxazolyl.

[0023] Examples of aryl and heteroaryl ring systems include thiazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each of the above noted aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. The terms "arylene" and "heteroarylene" refer to the divalent forms of aryl and heteroaryl, respectively.

[0084] For brevity, the term "aryl," when used in combination with other terms (e.g., aryloxy, arylthioxy, arylalkyl), includes both aryl and heteroaryl rings as defined above. Thus, the terms "arylalkyl" and "heteroarylalkyl" mean that an aryl or heteroaryl group includes a group bonded to an alkyl group (e.g., benzyl, phenethyl, pyridylmethyl, furylmethyl, etc.), and include alkyl groups in which a carbon atom (e.g., a methylene group) has been replaced with, for example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, etc.). However, as used herein, the term "haloaryl" is meant to encompass only aryls substituted with one or more halogens.

[0085] When a heteroalkyl, heterocycloalkyl, or heteroaryl contains a specific number of members (eg, "3 to 7 membered"), the term "member" refers to a carbon or heteroatom.

[0086] Furthermore, the formula: [ka] As used herein, a structure generally represented by the formula: refers to a ring structure, for example, but not limited to, aliphatic and / or aromatic rings, such as 3-carbon, 4-carbon, 5-carbon, 6-carbon, 7-carbon, etc., including saturated, partially saturated, and unsaturated ring structures, and includes substituent R groups, where the R groups may be present or absent, and if present, one or more R groups may each be substituted on one or more available carbon atoms of the ring structure. The presence or absence of an R group and the number of R groups is determined by the value of the variable "n," which is generally an integer having a value ranging from 0 to the number of carbon atoms on the ring that are available for substitution. Each R group, when present in multiple instances, is substituted on an available carbon of the ring structure rather than another R group. For example, the above structure, where n is 0 to 2, would encompass a class of compounds including, but not limited to: [ka] etc.

[0087] A dashed line representing a bond in a cyclic ring structure indicates that the bond may or may not be present within the ring, i.e., the dashed line representing a bond in a cyclic ring structure indicates that the ring structure is selected from the group consisting of saturated ring structures, partially saturated ring structures, and unsaturated ring structures.

[0088] symbol [ka] indicates the point of attachment of the moiety to the rest of the molecule.

[0089] When a named atom of an aromatic ring or heteroaromatic ring is defined as "absent," said named atom is replaced by a direct bond.

[0090] Each of the above terms (e.g., "alkyl," "heteroalkyl," "cycloalkyl," and "heterocycloalkyl," "aryl," "heteroaryl," "phosphonate," and "sulfonate," as well as their divalent derivatives) are meant to include both substituted and unsubstituted forms of the indicated radical. Optional substituents for each type of radical are provided below.

[0091] Substituents for alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl monovalent and divalent derivative groups (including groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be one or more of a variety of groups selected from, but not limited to, the following: -OR', ═O, ═NR', ═N-OR', -NR'R”, -SR', -halogen, -SiR'R"R'”, -OC(O)R', -C(O)R', -COR', -C(O)NR'R”, -OC(O)NR'R”, -NR"C(O)R', -NR'-C(O)NR"R'”, -NR"C(O)OR', -NR-C(NR'R")═NR'”, -S(O)R', -S(O)R', -S(O)NR'R", -NRSOR', -CN, and -NO (where m' is the total number of carbon atoms in such group). R', R", R'", and R"" can each independently represent hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g., aryl substituted with 1 to 3 halogens), substituted or unsubstituted alkyl, alkoxy or thioalkoxy group, or arylalkyl group. As used herein, an "alkoxy" group is an alkyl attached to the remainder of the molecule through a divalent oxygen. When a compound of the invention includes more than one R group, for example, each of the R groups is independently selected such that each R', R", R'", and R"" group is independently selected when two or more of these groups are present. When R' and R" are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example, -NR'R" is meant to include, but not be limited to, 1-pyrrolidinyl and 4-morpholinyl.From the above discussion of substituents, one of ordinary skill in the art will understand that the term "alkyl" is meant to include groups that contain carbon atoms bonded to groups other than hydrogen groups, such as haloalkyl (e.g., -CF and -CHCF) and acyl (e.g., -C(O)CH, -C(O)CF, -C(O)CHOCH, etc.).

[0092] Similar to the substituents described above for alkyl groups, exemplary substituents for the aryl and heteroaryl groups (and their divalent derivatives) are varied and are selected from, for example, halogen, —OR′, —NR′R″, —SR′, —SiR′R″R′″, —OC(O)R′, —C(O)R′, —COR′, —C(O)NR′R″, —OC(O)NR′R″, —NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)OR′, —NR—C(NR′R″R′”)═NR″”, —NR—C(NR′R”)═NR′″, —S(O)R′, —S(O)R′, —S(O)NR′R″, —NRSO═R′, —CN, and —NO, —R′, —N, —CH(Ph), fluoro(C1-C4)alkoxo, and fluoro(C1-C4)alkyl; where R', R", R'" and R"" may be independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl. When a compound of the invention includes more than one R group, for example, each of the R groups is independently selected, such that each R', R", R'" and R"" group is independently selected when two or more of these groups are present.

[0093] Two substituents on adjacent atoms of an aryl or heteroaryl ring are of the formula -TC(O)-(CRR') qAlternatively, two substituents on adjacent atoms of the aryl or heteroaryl ring may optionally form a ring of the formula -U-, where T and U are independently -NR-, -O-, -CRR'- or a single bond, and q is an integer from 0 to 3. Alternatively, two substituents on adjacent atoms of the aryl or heteroaryl ring may optionally form a ring of the formula -A-(CH2) r -B-, wherein A and B are independently -CRR'-, -O-, -NR-, -S-, -S(O)-, -S(O)2-, -S(O)2NR'- or a single bond, and r is an integer of 1 to 4.

[0094] One of the single bonds of the new ring thus formed may optionally be replaced with a double bond. Alternatively, two substituents on adjacent atoms of the aryl or heteroaryl ring may be a group of the formula -(CRR') s -X'-(C"R'") d wherein s and d are independently integers from 0 to 3, and X' is -O-, -NR'-, -S-, -S(O)-, -S(O)2-, or -S(O)2NR'-. The substituents R, R', R" and R'" can be independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.

[0095] As used herein, the term "acyl" refers to an organic acid group in which the -OH of the carboxyl group is replaced with another substituent and has the general formula RC(=O)-, where R is an alkyl, alkenyl, alkynyl, aryl, carbocyclic, heterocyclic, or aromatic heterocyclic group, as defined herein. As such, the term "acyl" specifically includes arylacyl groups such as 2-(furan-2-yl)acetyl- and 2-phenylacetyl groups. Specific examples of acyl groups include acetyl and benzoyl. Acyl groups are also intended to include amides, -RC(=O)NR', ​​esters, -RC(=O)OR', ketones, -RC(=O)R', and aldehydes, -RC(=O)H.

[0096] The terms "alkoxyl" or "alkoxy" are used interchangeably herein and refer to a saturated (i.e., alkyl-O-) or unsaturated (i.e., alkenyl-O- and alkynyl-O-) group attached to the parent molecular moiety through an oxygen atom, where the terms "alkyl," "alkenyl," and "alkynyl" are as previously defined, e.g., C 1-20 Inclusive straight, branched or cyclic, saturated or unsaturated oxo-hydrocarbon chains may be included, such as methoxy, ethoxy, propoxyl, isopropoxyl, n-butoxyl, sec-butoxyl, tert-butoxyl, and n-pentoxyl, neopentoxyl, n-hexoxyl, and the like.

[0097] The term "alkoxyalkyl" as used herein means an alkyl-O-alkyl ether, such as a methoxyethyl or ethoxymethyl group.

[0098] "Aryloxyl" means an aryl-O- group, where the aryl group is as defined above, including substituted aryl. As used herein, the term "aryloxyl" can refer to phenyloxyl or hexyloxyl, and alkyl, substituted alkyl, halo, or alkoxyl substituted phenyloxyl or hexyloxyl.

[0099] "Aralkyl" means an aryl-alkyl group, where aryl and alkyl are as previously described, including substituted aryl and alkyl. Exemplary aralkyl groups include benzyl, phenylethyl, and naphthylmethyl.

[0100] "Aralkyloxyl" means an aralkyl-O- group, where the aralkyl group is as previously described. An exemplary aralkyloxyl group is benzyloxyl, i.e., C6H5-CH2-O-. An aralkyloxyl group can be optionally substituted.

[0101] "Alkoxycarbonyl" means an alkyl-OC(=O)- group. Exemplary alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl, butyloxycarbonyl, and tert-butyloxycarbonyl.

[0102] "Aryloxycarbonyl" means an aryl-OC(=O)- group. Exemplary aryloxycarbonyl groups include phenoxy- and naphthoxy-carbonyl.

[0103] "Aralkoxycarbonyl" means an aralkyl-OC(=O)- group. An exemplary aralkoxycarbonyl group is benzyloxycarbonyl.

[0104] "Carbamoyl" means an amide group of the formula -C(=O)NH2. "Alkylcarbamoyl" means the group R'RN-C(=O)- where one of R and R' is hydrogen and the other of R and R' is alkyl and / or substituted alkyl as defined above. "Dialkylcarbamoyl" means the group R'RN-C(=O)- where each of R and R' is independently alkyl and / or substituted alkyl as defined above.

[0105] The term carbonyldioxyl, as used herein, refers to a carbonate group of formula -OC(=O)-OR.

[0106] "Acyloxyl" means an acyl-O- group where acyl is as previously described.

[0107] The term "amino" refers to the -NH group and also refers to nitrogen-containing groups known in the art derived from ammonia in which one or more hydrogen radicals have been replaced by an organic radical. For example, the terms "acylamino" and "alkylamino" refer to specific N-substituted organic radicals with acyl and alkyl substituents, respectively.

[0108] As used herein, "aminoalkyl" refers to an amino group covalently attached to an alkylene linker. More specifically, as used herein, the terms alkylamino, dialkylamino, and trialkylamino refer to one, two, or three alkyl groups, as defined above, respectively, attached to the parent molecular moiety through a nitrogen atom. The term alkylamino refers to a group having the structural formula -NHR', where R' is an alkyl group, as defined above; the term dialkylamino refers to a group having the structural formula -NR'R", where R' and R" are each independently selected from the group consisting of alkyl groups. The term trialkylamino refers to a group having the structural formula -NR'R"R"', where R', R", and R'" are each independently selected from the group consisting of alkyl groups. Additionally, R', R'' and / or R''' taken together optionally represent -(CH2) k - where k is an integer from 2 to 6. Non-limiting examples include methylamino, dimethylamino, ethylamino, diethylamino, diethylaminocarbonyl, methylethylamino, isopropylamino, piperidino, trimethylamino, and propylamino.

[0109] An amino group is -NR'R" where R' and R" are typically selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0110] The terms alkylthioether and thioalkoxyl refer to saturated (i.e., alkyl-S-) or unsaturated (i.e., alkenyl-S- and alkynyl-S-) groups attached to the parent molecular moiety through a sulfur atom. Non-limiting examples of thioalkoxyl moieties include methylthio, ethylthio, propylthio, isopropylthio, n-butylthio, and the like.

[0111] "Acylamino" means the group acyl-NH- where acyl is as previously described. "Aroylamino" means the group aroyl-NH- where aroyl is as previously described.

[0112] The term "carbonyl" refers to the group -C(=O)- and may include aldehyde groups of the general formula RC(=O)H.

[0113] The term "carboxyl" refers to a -COOH group. Such a group is also referred to herein as a "carboxylic acid" moiety.

[0114] As used herein, the terms "halo," "halide," or "halogen" refer to fluoro, chloro, bromo, and iodo groups. Additionally, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C1-C4)alkyl" is meant to include, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0115] The term "hydroxyl" refers to an --OH group.

[0116] The term "hydroxyalkyl" refers to an alkyl group substituted with an --OH group.

[0117] The term "mercapto" refers to the group --SH.

[0118] The term "oxo" as used herein means an oxygen atom that is double bonded to a carbon atom or other element.

[0119] The term "nitro" refers to the group --NO.sub.2.

[0120] The term "thio" refers to a compound described previously herein, wherein a carbon or oxygen atom has been replaced by a sulfur atom.

[0121] The term "sulfate" refers to the group -SO4.

[0122] The term thiohydroxyl or thiol as used herein means a group of formula -SH.

[0123] More specifically, the term "sulfide" refers to a compound having a group of formula -SR.

[0124] The term "sulfone" refers to a compound having a sulfonyl group -S(O2)R.

[0125] The term "sulfoxide" refers to a compound that has a sulfinyl group -S(O)R.

[0126] The term ureido means a urea group of formula -NH-CO-NH2.

[0127] With respect to compounds of Formula (I), the term "protecting group" refers to a chemical substituent that can be selectively removed by readily available reagents that do not attack the regenerated functional group or other functional groups in the molecule. Suitable protecting groups are known in the art and continue to be developed. Suitable protecting groups are described, for example, in Wutz et al. ("Greene's Protective Groups in Organic Synthesis, 4th Edition," Wiley-Interscience, 2007). Protecting groups for the protection of carboxyl groups, as described by Wutz et al. (pp. 533-643), are used in some embodiments. In some embodiments, the protecting group is removable by acid treatment. Representative examples of protecting groups include, but are not limited to, benzyl, p-methoxybenzyl (PMB), tert-butyl (t-Bu), methoxymethyl (MOM), methoxyethoxymethyl (MEM), methylthiomethyl (MTM), tetrahydropyranyl (THP), tetrahydrofuranyl (THF), benzyloxymethyl (BOM), trimethylsilyl (TMS), triethylsilyl (TES), t-butyldimethylsilyl (TBDMS), and triphenylmethyl (trityl, Tr). One of skill in the art will recognize appropriate situations requiring protecting groups and will be able to select an appropriate protecting group for use in a particular situation.

[0128] Throughout the specification and claims, a given chemical formula or name encompasses all tautomers, homologs and optical isomers and stereoisomers, as well as racemic mixtures where such isomers and mixtures exist.

[0129] Certain compounds of the present invention may possess asymmetric carbon atoms (optical or chiral centers) or double bonds; enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomers, which may be defined by absolute stereochemistry as (R)- or (S)-, or D- or L- of amino acids, and individual isomers are encompassed within the scope of the present invention. The compounds of the present invention do not include compounds known in the art as being too unstable to synthesize and / or isolate. The present invention is meant to include compounds in racemic, scalemic, and optically pure form. Optically active (R)- and (S)- or D- and L-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When compounds described herein contain olefinic bonds or other centers of geometric asymmetry, unless otherwise specified, the compounds are intended to include both E and Z geometric isomers.

[0130] Unless otherwise stated, structures depicted herein are also meant to include all stereochemical forms of said structure (i.e., R and S configurations for each asymmetric center). Accordingly, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the invention.

[0131] It will be apparent to one of ordinary skill in the art that certain compounds of the present invention may exist in tautomeric forms, and all such tautomeric forms of the compounds are within the scope of the present invention. As used herein, the term "tautomer" means one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to another.

[0132] Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms, for example, the replacement of a hydrogen by deuterium or tritium, or 13 C- or I4 Compounds having the present structures with the replacement of a carbon with a C-enriched carbon are within the scope of this invention.

[0133] The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may contain unnatural proportions of atomic isotopes, such as tritium ( 3 H), iodine-125( 125 I) or carbon-14 ( 14 C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.

[0134] The compounds of the present invention may exist as salts. The present invention includes such salts. Examples of applicable salt forms include hydrochloride, hydrobromide, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate (e.g., (+)-tartrate, (-)-tartrate, or a mixture thereof, including a racemic mixture), succinate, benzoate, and salts of 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 a compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid, either directly or in a suitable inert solvent, or by ion exchange. Examples of acceptable acid addition salts include salts 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, as well as 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-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. Also included are salts of amino acids such as alginate, and salts of organic acids such as glucuronic acid or galacturonic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups that allow the compounds to be converted into either base or acid addition salts.

[0135] The neutral forms of the compounds can be 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.

[0136] Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to the unsolvated forms and are included within the scope of the present invention. Certain compounds of the present invention may exist in polycrystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[0137] In addition to salt forms, the present invention provides compounds in prodrug form. Prodrugs of the compounds described herein are compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention. Additionally, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of the present invention when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0138] Following long-standing patent law convention, the terms "a," "an," and "the" mean "one or more" when used in this application, including the claims. Thus, for example, reference to "a subject" includes a plurality of subjects, etc., unless the context clearly dictates otherwise (e.g., a plurality of subjects).

[0139] Throughout this specification and claims, the terms "comprise," "comprises," and "comprising" are used in a non-exclusive sense unless the context requires otherwise. Similarly, the term "include" and its grammatical variations are intended to be open-ended, and the recitation of items in a list does not exclude other similar items that may be substituted for or added to the listed items.

[0140] For purposes of this specification and the appended claims, unless otherwise indicated, all numbers expressing quantities, sizes, dimensions, proportions, shapes, dosage forms, parameters, percentages, amounts, characteristics, and other numerical values ​​used in the specification and claims are to be understood as being modified in all instances by the term "about," even if the term "about" does not explicitly appear in conjunction with that value, amount, or range. Thus, unless indicated to the contrary, the numerical parameters set forth in the following specification and appended claims are not, and need not be, exact, but may be approximated and / or larger or smaller, as desired, depending upon the desired properties sought to be obtained by the present subject matter, while taking into account tolerances, conversion factors, rounding, measurement error, and the like, as well as other factors known to those of ordinary skill in the art. For example, when referring to a value, the term "about" is meant to encompass, in some embodiments, ±100%, in some embodiments ±50%, in some embodiments ±20%, in some embodiments ±10%, in some embodiments ±5%, in some embodiments ±1%, in some embodiments ±0.5%, and in some embodiments ±0.1% variations from the particular amount, where such variations are appropriate for practicing the methods and using the compounds of the invention.

[0141] Furthermore, the term "about," when used in connection with one or more numbers or numerical ranges, should be understood to mean all such numbers, including every number within a range, modifying that range by extending the boundaries above and below the stated numerical values. Reciting numerical ranges by endpoints includes all numbers encompassed within that range, e.g., whole integers, fractions thereof (e.g., reciting 1 to 5 includes 1, 2, 3, 4, and 5, as well as fractions thereof (e.g., 1.5, 2.25, 3.75, 4.1, etc.), and any range within that range. [Example]

[0142] The following examples are included to provide guidance to one of ordinary skill in the art in carrying out representative embodiments of the inventive subject matter. In light of the present invention and the general level of skill of one of ordinary skill in the art, one of ordinary skill in the art will recognize that the following examples are merely illustrative, and that many variations, modifications, and alterations may be made without departing from the scope of the inventive subject matter. The following synthetic descriptions and specific examples are intended for illustrative purposes only and should not be construed as limiting the methods for preparing the compounds of the present invention by other methods.

[0143] Example 1 overview The use of PSMA-conjugated ureas conjugated to chelated radiometals via various linking groups to enable imaging and radiotherapy of PSMA-expressing tumors has already been reported in several patent applications and publications (Tykvart et al. (2015) Journal of Medicinal Chemistry 58, 4357-63; Banerjee et al. (2015) Journal of Nuclear Medicine 56, 628-34; Benesova et al. (2015) Journal of Nuclear Medicine 56, 914-20; Weineisen et al. (2014) EJNMMI Res 4, 1-15; WO 2009002529 A2; WO 2009070302 A1). A new class of high-affinity binders has been prepared by modifying the urea linker at the ε-amine position with a p-Br-benzyl group. The structures of the compounds of the present invention are shown in Figure 1.

[0144] Without wishing to be bound by any particular theory, it is believed that when the epsilon-amino group of the lysine in the Lys-Glu-urea moiety is modified with a p-Br-benzyl group, radiometal-chelated Glu-lysine urea-based theranostic agents targeting prostate-specific membrane antigen (PSMA) exhibit high binding affinity for PSMA and high uptake in PSMA-expressing tumors, and low renal uptake in standard mouse models of prostate cancer. According to one embodiment, 177 Lu-1 demonstrated significant radiotherapeutic efficacy in mice bearing PSMA+ PC3 tumors, with approximately 50% remission.

[0145] Example 2 Materials and Methods Chemical Synthesis of 8. The synthesis of compound 1 is described in Scheme 1. Bromobenzaldehyde (121.0 mg, 0.654 mmol) was slowly added to a stirred solution of Boc-protected urea 4 (300.0 mg, 0.615 mmol) in 5 ml of methanol in an ice bath and allowed to warm to room temperature. After 1 h, sodium cyanoborohydride (158.0 mg, 2.5 mmol) was added and the reaction was left stirring overnight. The crude reaction mixture was evaporated, redissolved in dichloromethane, and purified by normal-phase silica chromatography (95:5, methylene chloride:methanol) and dried in vacuo to give 5 in good yield. Yield: 80%. ESI-MS: 656.56 [M+H] + , Found: 656.5. TSTU (32.6 mg, 108 μmol), Boc-5-aminovaleric acid (23.5 mg, 108 μmol), and DIPEA (37.7 μL, 216 μmol) were dissolved in 300 μL of DMF and stirred at room temperature. After 1 h, compound 5 (71.0 mg, 108 μmol) was added and rinsed three times with DMF (50 μL each). The reaction mixture was stirred for 4 h and stored at 4 °C overnight. The crude reaction mixture was C 18Purification was carried out by semi-preparative HPLC on a column (40% water (0.1% TFA) / 60% ACN (0.1% TFA) / 5 min, 60-90%, over 20 min). t 21 min. The purified fractions were combined, evaporated, and dried under high vacuum for 10 min. ESI-MS: 572.44 [M+H]+, found: 572.4. Compound 6 was dissolved in dichloromethane (1.5 mL) and cooled in an ice bath. After equilibration, TFA (1.5 mL) was added, and the mixture was stirred for 3 h, warming to room temperature during the process. The mixture was dried under a stream of nitrogen, dissolved in water, and lyophilized to give 31.8 mg of compound 7. Yield: 54 μmol, 54%. To a stirred solution of 6 (12.2 mg of TFA salt) and DIPEA (15.2 μL, 87.0 μmol) in DMSO (130 μL) equilibrated to 40 °C, p-SCN-bn-DOTA (12.2 mg, 17.7 μmol) was added. The reaction mixture was stirred at 40 °C for 4 hours and stored at 4 °C overnight. The reaction mixture was purified by reverse-phase HPLC (20% ACN, 5 min hold, then 20-40% over 19 min). Rt approx. 12 min. The purified fractions were combined, rotary evaporated to reduce the volume, and then lyophilized. ESI-MS: 1138.37 [M+H] + Found: 1138.5. Compound 1 was further purified by HPLC using a gradient method comprising a mobile phase of 88% water (containing 0.1% TFA) and 22% CHCN (0.1% TFA) from 1 to 5 minutes, followed by 88% water (containing 0.1% TFA) and 12% CHCN (0.1% TFA) from 0 to 5 minutes, and a gradient from 88% water to 44% water and 12% acetonitrile to 56% acetonitrile from 5 to 25 minutes at a flow rate of 8 mL / min.

[0146] Chemical synthesis of 2. This compound was synthesized using the same intermediate 7 and coupled to commercially available DOTA-NHS ester. ESI-MS: 974.86 [M+H] + , Actual value: 974.5.

[0147] Chemical synthesis of 3. This compound was synthesized using intermediate 4 and coupled with commercially available Boc-5-aminovaleric acid and DOTA-NHS ester. ESI-MS: 970.05 [M+H] + , Actual measured value: 970.1.

[0148] 177 Radiolabeling of Lu-1. 1.0 μl of 0.1 N HCl 177 LuCl (1 mCi) was added to 70 μl of NH4OAc buffer (0.2 M, pH 4) and made up to 5 μl of 2 mM in 0.2 M NH4OAc. The pH of the mixture was approximately 4.0. The mixture was kept at 80 °C for 1 hour and purified by HPLC. The HPLC method consisted of a mobile phase of 77% water (containing 0.1% TFA) and 23% CH3CN (0.1% TFA) from 1 to 5 min, followed by a gradient consisting of 77% to 57% water and 23% to 43% acetonitrile from 5 to 25 min; 5% to 5% water and 95% to 95% acetonitrile from 25.01 to 30 min; and 77% to 77% water and 23% to 23% acetonitrile from 30.01 to 37 min. The flow rate was 1.0 ml / min; λ: 200 nm; and a C8 column (25 × 4.6 mm), Varian microsob-MV 100-5. Radiolabeled chromatograms were analyzed. 177 Lu-1 eluted between 17.1 and 20 min, whereas the unlabeled chelator eluted between 21 and 22 min.

[0149] The HPLC method comprises: 177 Lu-1 and 177The HPLC method used to prepare Lu-7 was a gradient method consisting of a mobile phase of 88% water (containing 0.1% TFA) and 22% CH3CN (0.1% TFA) from 1 to 5 min, followed by a gradient of 88% to 75% water and 12% to 25% acetonitrile from 5 to 27 min; 5% to 5% water and 95% to 95% acetonitrile from 27.01 to 32 min; and 88% to 18% water and 12% to 22% acetonitrile from 32.01 to 37 min. Flow rate: 1.0 ml / min; λ: 200 nm; C8 column (25 x 4.6 mm), Varian microsob-MV 100-5. Radiolabeling 177 Lu-2 eluted between 13.1 and 15.0 min, and the unlabeled chelator eluted between 16 and 17 min. 177 Lu-3 eluted from 13.1 to 15.0 min, unlabeled chelators eluted from 10 to 12 min and 18 to 20 min, and unlabeled drugs eluted from 14 to 16 min.

[0150] Scheme 1. Synthesis of Compound 1 [ka] a. 4-Bromobenzaldehyde, NaBH3CN, MeOH, 1% acetic acid; b. BocNH(CH2)4CO2H, HATU, DIEA, DMF; c. TFA / CH2Cl2; d. DOTA-Bn-SCN, DMSO, DIEA

[0151] Example 3 Results and Discussion Chemical and Radiochemical Synthesis and Characterization. Following literature procedures (Tykvart et al. (2015) Journal of Medicinal Chemistry 58, 4357-63), a p-bromobenzyl-modified Glu-Lys urea (2) was prepared in good yield by reductive alkylation of 2 with p-bromobenzaldehyde in the presence of sodium cyanoborohydride in methanol to give 4. The small aliphatic linker, Boc-5-aminovaleric acid, was coupled to the same ε-Lys amine of 4, followed by removal of the BOC group and conjugation of 6 with commercially available DOTA-Bn-SCN to give 1 in moderate yield. Compound 2 was synthesized by coupling with the same intermediate 6 using DOTA-NHS ester as the chelating agent. Compound 3 was synthesized as a control agent without any p-bromobenzyl group. All three compounds were synthesized in good yield and purity in ammonium acetate buffer at 80 °C and pH 4. 177 The binding affinities of the novel compounds are shown in Table 1. Both 1 and 2, modified with p-bromo-benzyl groups, showed higher binding affinities compared to 3.

[0152] [Table 1]

[0153] Cell-binding properties. 177 Lu agents were further evaluated in cells and animals using standard syngeneic cell lines PSMA+ PC3 PIP and PSMA-negative PC3 flu cells. 177 Lu-1 and 177 Both the Lu-2 177 It showed higher uptake in PSMA+ PC3 cells compared to Lu-3. Furthermore, internalization studies showed 177 Lu-1 177The results showed that the internalization activity of these drugs was nearly twice as high as that of Lu-3. All three drugs showed significantly lower uptake in PSMA-negative PC3 flu cells. 177 Lu-1 was further evaluated for therapeutic efficacy in clonogenic assays and compared favorably with the previous lead compound SR6 (Banerjee et al. (2015) Journal of nuclear medicine 56, 628-34) and 177 Lu-PSMA-617 (Benesova et al. (2015) Journal of nuclear medicine 56, 914-20) and 177 Lu was compared with drugs in clinical trials, including PSMA-I&T (Weineisen et al. (2014) EJNMMI Res 4, 1-15). 177 Lu-1 was able to produce approximately 100% cell killing in PSMA+ PC3 PIP cells using a 10 μCi dose, but no significant toxicity was observed against PSMA- PC3 flu cells.

[0154] [Table 2]

[0155] Biodistribution. 177 Lu-1 and 177 In vivo tissue biodistribution studies were performed on Lu-2. The results are listed in Tables 3 and 4. Uptake in PSMA+ PC3 PIP tumors was 177 Lu-1 is 177 It showed significantly higher uptake and retention than Lu-2. 177 Lu-2 agents are 177 The compounds of the present invention exhibited a 5-fold lower kidney uptake than Lu-1, and as shown in Figure 3, the tumor / kidney uptake of the compounds of the present invention was significantly higher than that of the previous lead compound. 177 Lu-SR6, 177 Lu-PSMA-617 and 177 Compared with that of Lu-PSMA-I&T. 177The PSMA+ PC3 PIP tumor-to-kidney ratio for Lu-2 was 177 It showed higher tumor uptake and retention than Lu-1. 177 Lu-1 was further evaluated for its theranostic efficacy (imaging and therapeutic effects) in a pilot study using a small group of animals.

[0156] [Table 3]

[0157] [Table 4]

[0158] Small animal SPECT imaging and treatment effects. Figure 4 shows the results of treatment over the course of the study, from 1 to 8 days post-injection. 177 SPECT imaging of Lu-1 is shown. Mice (n = 10) bearing PSMA+ PC3 PIP tumors (3-5 mm in size) were administered a single dose of 3 mCi via tail vein injection. Another group of mice (n = 10) received saline as a control. Mice were monitored twice weekly by weight and tumor size. Mice in the control group were euthanized after 4 to 8 weeks due to tumor size exceeding 12 mm. In the treatment group, tumors were completely eradicated in 50% of mice. These mice initially lost weight and regained their original weight after 2 weeks. The results are shown in Figure 5. Figures 6A and 6B show the 177 The therapeutic efficacy of Lu-1 (reduced tumor volume) was demonstrated compared to a saline control group. Five mice experienced complete disease remission and survived for more than five months.

[0159] In summary, when modified with a p-Br-benzyl group on the lysine epsilon-amino group of the Lys-Glu-urea moiety, radiometal-chelated Glu-lysine urea-based theranostic agents targeting prostate-specific membrane antigen (PSMA) showed binding affinity to PSMA and high uptake in PSMA-expressing tumors, with low uptake in the kidney in a standard mouse model of prostate cancer. 177 Lu-1 demonstrated significant radiotherapeutic efficacy in mice bearing PSMA+ PC3 tumors, with approximately 50% remission.

[0160] Example 4 PSMA-based high-LET agents for prostate cancer SUMMARY. In some embodiments, the present subject matter relates to optimizing the pharmacokinetics of radiometal-based drugs that target PSMA. Prior work in this field is described in International PCT Patent Application Publication Nos. WO 2009 / 002529 A2 and WO 2010 / 108125 A2, each of which is incorporated by reference in its entirety. More particularly, in some embodiments, the present subject matter relates to, but is not limited to, 213 Bi(t 1 / 2 46 min, E mean 8.4 MeV), 212 Pb(t 1 / 2 10.6 h, E mean 7.8 MeV), and 225 Ac(t 1 / 2 10 d, E mean We provide PSMA-targeted low molecular weight (LMW) theranostic agents labeled with α-emitting radiometals, including 6 MeV, 4α, with the goal of reducing off-target effects that currently prevent widespread adoption of this promising therapy.

[0161] The rationally designed alpha emitters and methods of use of the present invention were developed through detailed consideration of pharmacokinetics (PK), the physical half-life of the radiometal, and by receptor blocking to reduce uptake by normal tissues. Using alpha camera imaging guidance, sub-organ localization (hot spots) and their blocking were rapidly identified prior to therapeutic studies.

[0162] Thus, in some embodiments, the present invention includes radiometal-chelating Glu-lysine urea-based theranostic agents that target prostate-specific membrane antigen (PSMA) and are modified with a p-Br-benzyl group on the ε-amino group of the lysine in the Lys-Glu-urea moiety. These compounds exhibit higher binding affinity to PSMA, significantly reduced renal uptake in standard mouse models of prostate cancer, and high tumor uptake in PSMA-expressing tumors. These optimized agents are also suitable for the treatment of highly toxic radiometals. 213 Bi, 212 Pb, and 225 Radiolabeled with Ac.

[0163] example [ka]

[0164] The ligands were synthesized as described above. The following compounds may also be synthesized by the method of the present invention: [ka]

[0165] 213 Bi-1 / 225 Ac-1 / 203 Radiolabeling of Pb-1. All radiolabeled compounds were 213 It was prepared according to the general method described for Bi-1. 213 Bi was produced by Oak Ridge National Laboratory 225 Ac / 213 Bi- eluted from the generator. Freshly eluted 213 10 μg of Bi (18.2 MBq) was added to the 1st solution, and the solution was adjusted to 4 to 5 with 3M NH4OAc. This solution was heated in a microwave oven at 95 °C and 40 watts for 5 minutes. Next, 10 μL of 1 mM Na-DTPA solution was added to the 1st solution, which did not contain the complex. 213 Added to Bi. 213 The specific activity of Bi-1 was > 7.4 MBq / μg in all experiments. 213 Bi-1 was treated with Phenomenex C 18 Luna 10×250 mm 2Purification was performed using a column and a Varian Prostar System (Palo Alto, CA) equipped with a Varian ProStar 325 UV-Vis variable wavelength detector and a Bioscan (Poway, CA) flow-count inline radioactivity detector, all controlled by Galaxie software. The flow rate was 1 mL / min using water (0.1% TFA) (A) and CH3CN (0.1% TFA) (B) as eluents. To ensure uniform purity, an isocratic solution of 80% A and 20% B was used to separate excess ligand from the radiolabeled compound. Specific radioactivity was calculated as the ratio of the radioactivity eluting at the product retention time during preparative HPLC purification to the mass corresponding to the area under the UV absorbance curve. The purity of the test compound was >95% as determined by analytical HPLC using absorbance at 254 nm. The yield of radiolabeled compound was also assessed using silica gel instant TLC (ITLC) with 0.9% sodium chloride as the mobile phase. The radiolabeled sample was diluted with 10 mM diethylenetriaminepentaacetic acid (DTPA) at a pH of approximately 4. Three microliters of the diluted sample was spotted onto an ITLC silica gel strip and developed in the chromatography chamber. After migration to the solvent front was complete, the ITLC sample strip was dried, cut in half, and counted in a Wallac Wizard γ-counter (PerkinElmer, Boston, MA) to determine the yield of radiolabeled product. Radiochemical purity was assessed by high-performance liquid chromatography (HPLC) analysis.

[0166] Figures 7, 8, 9, 10, 11 and 12 show the in vitro and in vivo characterization. 225 The biodistribution of Ac-1 is shown in Table 5. [Table 5]

[0167] Development of receptor blockade for normal organs. Blocking effect of 1 177 Lu-1 was used to quantify normal tissues. A dramatic decrease in kidney uptake was observed, while high uptake in PSMA(+) tumors was maintained (Figure 12). Although a decrease in salivary gland uptake was observed in these blocking studies, the results were not statistically significant. This is because 177 This is likely due to high experimental error associated with very low Lu-1 uptake (< 0.5% at 2 h).

[0168] Blocking strategies. All blocking tests are performed on long-lived, decaying daughter nuclides. 213 Bi and 221 Considering the toxicity caused by Fr, it has the best performance. 225 Ac-labeling agents (e.g., 225 Ac-1). Once formed, these daughter nuclides are less likely to associate with chelating constructs due to their high atomic recoil energy as a result of alpha decay. Without wishing to be bound by any particular theory, 225 Acute salivary gland toxicity due to Ac-PSMA-617 was most likely due to the release of daughter nuclides in the striatal ducts or acini after internalization of the PSMA-based agent. Both biodistribution data and alpha camera imaging were used to interpret the data. To avoid kidney and salivary gland radiation toxicity, various pharmacological agents were evaluated. In addition to com-injection, a 15-minute blockade of 30 minutes prior to injection was examined to determine whether any significant changes occurred due to the short plasma half-life of the agent. Effective blockade agents were combined in two or three doses to determine whether any improvements could be achieved with the combined agent.

[0169] The following experiments can be carried out within the scope of the present invention:

[0170] Receptor blocking (self-blocking) tests, e.g. 225 free ligand 1 of Ac-1 (as in Figure 12 );

[0171] in PSMA-expressing normal tissues to provide synergistic receptor blockade and metal ion sequestration 213 Competitive blocking using dithiol chelators that scavenge Bi;

[0172] 2,3-Dimercapto-1-propanesulfonic acid (DMPS) 213 Use of dithiol chelating agents to remove Bi (because dithiol chelating agents have been shown to be effective in various animal and human studies). 213 (Because it has been shown to enhance the excretion of Bis or other heavy metals);

[0173] Francium (Fr) is an alkali metal, like sodium and potassium, and is therefore an important transporter for acinar cell fluid and electrolyte secretion, Na + / K + It can be absorbed by salivary gland ducts via the transporter NKCC1 / 2Cl- cotransporter.

[0174] Therefore, as reported, the FDA-approved diuretics furosemide and bumetanide can be used to reduce Na + and K. + By preventing reabsorption in the same way, 221 It can promote the removal of Fr; and

[0175] As reported, bismuth subnitrate suspension (100 mg / kg) induces competitive metal sequestering in renal tubular cells. 213 Bi or cold Bi +3 The effectiveness of competitive metal blockade of either the uptake of or its binding site (metallothionein-like protein) can be assessed.

[0176] Example 5 For PSMA-based radiopharmaceutical therapy, 177 Lu-labeled low molecular weight compounds

[0177] 5.1 Overview Prostate-specific membrane antigen (PSMA) is an important target for radionuclide therapy to treat metastatic castration-resistant prostate cancer. One objective of the present invention is to develop an optimized agent using low linear energy transfer irradiation of beta particles for targeted radionuclide therapy of metastatic castration-resistant prostate cancer. To this end, a novel PSMA-based 177 Lu-labeled radioligands were synthesized and evaluated for in vitro binding affinity and in vivo tumor targeting. Radiolabeling of new ligands was performed with high (>98%) radiochemical yield and specific activity. Cellular uptake and internalization data demonstrated specific uptake in PSMA(+)PC3 PIP cells for all drugs. Clonogenic cell viability assays were performed on selected drugs. Significantly reduced survival of PSMA(+)PC3 PIP cells was observed only after 48 hours of incubation compared with PSMA(-)PC3 flu cells. Selected compounds were evaluated for in vivo pharmacokinetics and therapeutic efficacy in a primary prostate cancer model, similar to clinical drugs. 177 Lu-PSMA-617 and 177 Further evaluation was performed in a head-to-head study with Lu-PSMA-I&T. Biodistribution data showed comparable tumor uptake in PSMA-expressing PC3 PIP tumors by 72 hours for the novel agents. These agents also demonstrated efficient tumor regression at 8 weeks after intravenous administration of 111 MBq (3 mCi) compared to untreated mice (n = 10). Representative compounds, 177 Lu-L1 showed a significantly improved survival rate. Renal autopsy was performed after 8 weeks of treatment with the selected drugs in tumor-bearing mice and after 1 year of treatment in tumor-free mice ( 177 Lu-L1, 111 MBq), and no radiation nephropathy was observed.

[0178] 5.2 Background Prostate-specific membrane antigen (PSMA), also known as glutamic acid carboxypeptidase II [GCPII] or N-acetyl-l-aspartyl-l-glutamic acid peptidase I [NAALADase I], is a type II cell surface metalloenzyme that has proven to be a useful clinical biomarker for prostate cancer. 1, 2 Approximately 80% or more of prostate tumors, as well as other solid tumors, show strong PSMA expression in newly formed blood vessels, mimicking tumor-associated angiogenesis. Lower levels are found in physiologically normal tissues (e.g., kidney, salivary gland, and small intestine). Because of the differential expression of PSMA in prostate tumors, clinicians and radiochemists have explored the use of PSMA as a target for the delivery of a wide range of diagnostic and therapeutic radionuclides using PET, SPECT, nanoparticles, optical agents, etc. PSMA-based low-molecular-weight PET imaging agents, such as 68 Ga-PSMA-11 3 and 18 F-DCFPyL 4 has revolutionized the early diagnosis of prostate cancer in men and is likely to undergo the FDA New Drug Application process in the next few years. 177 Lu-PSMA-617 and 177 Lu-PSMA-I&T 5-11 (Table 6) and halogens 131 I-MIP1095 2, 12, 13 Although the therapeutic potential of radionuclide therapy is promising, the safe and effective administration required for radionuclide therapy requires further investigation. For example, grade 1-3 hematological toxicities include xerostomia and mucositis. 13 Along with strong accumulation in the salivary glands causing 131 Reported in I-MIP1095. 225 Ac-based LMW agents are also in clinical trials and are being investigated for the treatment of salivary glands. 14Similar complications have been seen associated with PSMA-based radionuclide therapy. Prospective data collected in well-designed clinical trials are still lacking to address the long-term nephrotoxicity of PSMA-based radionuclide therapy, which is a major safety concern for these LMW radiotherapy agents.

[0179] The pharmacokinetics of drugs with high tumor-to-background ratios for safe and effective clinical applications (Figure 13) have been investigated for both imaging and treatment of prostate cancer. The overall biological profile of these drugs has been determined not only by receptor-specific binding but also by nonspecific interactions, which may be related to molecular weight, charge, hydrophilicity, and metabolic stability. Thus far, two broad classes of high-affinity drugs (K i < 20 nM) emerged from our studies: (i) Type I agents. These agents have high PSMA-specific tumor uptake and long retention in the body, but also show high uptake in many PSMA-expressing normal tissues, including the renal cortex and spleen. 15 For example: 99m Tc-oxo 16 、 64 Cu-1,4,7-triazacyclononane-1,4,7-2-triacetic acid (NOTA) 17 (Figure 13A), 68 Ga-PSMA-11 18 、 and drugs 68 Ga- / 177 Lu-PSMA-I&T, second generation) (Figure 13B), which have been administered to patients in several clinical trials. 6、7、19、20 These agents are generated by simply modifying the chelator or linker on the original linker / urea construct. 21 Recently, albumin-binding p-(iodophenyl)butyric acid moieties have been developed, leading to improved affinity for PSMA. 22-25Using this method, several high-affinity radioligands have been developed that exhibit high tumor retention, but also result in long and high radiation doses in the mouse renal cortex and other normal organs. (ii) Type II agents: These agents exhibit high tumor uptake and retention and rapid clearance from most normal tissues, including the kidney and salivary glands, resulting in high tumor-to-background ratios. 16、 17 。 Type II drugs include those developed by the present inventors (Figure 13A). 26 and PSMA-617 5 Drug II was recently investigated in a first-in-man study (Figure 13B). 27 A similar pattern of faster renal clearance from PSMA-expressing kidneys than from PSMA+ tumors was observed, likely related to faster flow through the normally organized renal vasculature compared with the relatively disorganized vasculature of the xenografts. 26, 28, 29 .

[0180] The subject matter of the present invention includes, in part, the preclinical evaluation of a novel series of small molecule compounds targeting PSMA to develop targeted theranostic radiopharmaceutical therapeutics for the treatment of metastatic prostate cancer. 125 I-iodo-benzoylamino)-pentyl]-ureido]-pentanedioic acid ( 125 Since I-DCIBzL) showed sustained tumor uptake for at least 48 hours in human PC xenografts, a 4-halobenzyl derivative of Lys-urea-Glu was investigated among other urea-based candidates. The agent significantly reduced tumors due to its high linear energy transfer and short-range emission of Auger electrons (<10 mm). 30 DCIBzL is one of the most potent PSMA binders known in the art (K i =0.01 nM). Furthermore, it emits alpha particles. 211 At-labeled DCIBzL showed significant therapeutic effects in both flank tumor and micrometastasis models. 31Therefore, the metal-based radiotherapy agents of the present invention were designed to contain halo-benzyl-urea-Glu derived from the structure of DCIBzL with some rational modifications to the linker and chelator to improve the binding affinity and pharmacokinetics of the agents. Several representative compounds were synthesized and the tumor targeting and pharmacokinetic properties of the agents were evaluated. This approach provides a promising candidate for PSMA-based targeted radionuclide therapy. 177 Lu, 212 Pb or 225 Ac can be made into an optimized drug with reduced off-target effects.

[0181] 5.3 Test Results 5.3.1 Synthesis Scheme. As shown in Figure 13, a series of representative compounds were synthesized for structure and activity relationship (SAR) studies based on the high-affinity drug DCIBzL. All ligands were designed to contain a (Br / I)-benzylLys-urea-Glu targeting moiety, except for ligands L8 and L14, which were designed as control agents. Ligands L1-L4, L7-L9, and L13 were designed to have short, flexible linkers, while L12 contains a rigid aromatic linker. In contrast, ligands L5, L6, and L14 were designed to have longer linkers, similar to previous lead drugs. 26 The effect of chelators was investigated in ligands L7 and L8 (by replacing the chelator DOTA-monoamide with DOTA-Bn-SCN as in L1) and in L9, which uses the DOTAGA chelator. Ligands L10 and L11 were designed with a rigid cyclohexyl linker as in PSMA-617, with DOTA-monoamide and DOTAGA chelator, to check the effect of rigid linkers and chelators compared to L1 and L9, respectively. Ligand L13 replaced Br-benzylLys-urea-Glu with Br-benzylLys-urea-Glu, a clinical PET imaging agent. 18The F-DCFPYL derivative was designed to have a different targeting moiety by replacing the Br-pyridylLys-urea-Glu with 32 Ligand L14, as recently studied by several research groups, 22-25, 33 We engineered a previously reported PSMA-binding targeting platform with an albumin-binding moiety, 4-(p-iodophenyl)butyrate, and investigated the effect of 4-(p-iodophenyl)butyrate on biodistribution properties.

[0182] All compounds were synthesized using solution-phase chemistry based on well-established methods for preparing Glu-Lys urea derivatives and associated linker and chelator conjugation chemistries. Ligands L1 and L2 were synthesized as described in Scheme 2. [ka]

[0183] Scheme 2. a Reagents and conditions: (a) NaBHCN, MeOH; (b) Boc-5-aminovaleric acid TSTU, DIPEA, DMF or Boc-6-aminohexanoic acid N-hydroxysuccinimide ester, EtN, DMF, overnight, room temperature; (c) 50% TFA / CHCl; c) 3a or 3c, DOTA-monoamide-NHS, DIPEA, DMSO, room temperature, 3 hours; (d) 177 Lu +3 / Lu +3, pH ~4-50.2 NH4OAC, 5 min, 95°C microwave.

[0184] The urea-lysine intermediate di-tert-butyl(((S)-1-(tert-butoxy)-6-((4-iodo / bromobenzyl)amino)-1-oxohexan-2-yl)carbamoyl)-L-glutamate, 1a or 1b (5-X-Bn)-Lys-urea-Glu (X = Br / I), was prepared according to literature methods. 34was synthesized with some modifications. Using these two intermediate compounds, L3, L4, L5, L6, L8, L9, L10, L11, and L12 were also synthesized. Briefly, Boc-5-aminovaleric acid was coupled to 1a and 1b to give 2a and 2c, followed by simultaneous removal of the tert-Butyl and N-Boc groups to generate 3a and 3c, respectively, in over 90% yield. Compounds 3a and 3c were then reacted with the N-hydroxosuccinimide (NHS) ester of DOTA monoamide to give the target ligands L1 and L2 in high yield. Ligands L3 and L4 were synthesized as shown in Scheme 3. [ka]

[0185] Scheme 3. a Reagents and conditions: (a) DIPEA, DMF, room temperature, 16 hours; (b) TSTU, TEA, DMSO, 4 hours; (c) 50% TFA / CH2Cl2; (d) 3b or 3d, DIPEA, DMSO.

[0186] The coupling reaction was carried out using the rigid linker p-aminomethylbenzoic acid and DOTA-monoamide NHS ester to give compound 4 in quantitative yield. Compound 5 was then obtained by treating compound 4 with TSTU in the presence of trimethylamine, followed by removal of the tert-butyl group using TFA / CHCl to generate reactive intermediate 6 in high yield. Ligands L3 and L4 were obtained in good yield after simple coupling reactions between 6 and 3b and 3d. Ligands L5 and L6 were synthesized according to the synthetic route shown in Scheme 4. [ka]

[0187] Scheme 4. a Reagents and conditions: (a) DSS, TEA, DMF; (b) 50% TFA / CH2Cl2; (c) 4, DIPEA, DMSO, 3 h.

[0188] First, compound 7 was synthesized using the previously reported 35 Compound 1a was synthesized by reacting 1a with disuccinimidyl suberate (DSS) as shown. Compound 7 was then treated with 50% TFA / CHCl to remove the t-butyl group, yielding compound 8, which was then coupled with Boc-5-aminovaleric acid, followed by removal of the Boc group and coupling with compound 6 to yield L5. Furthermore, following the synthetic route described in Scheme 2, three analogs of L1 were synthesized by replacing DOTA-monoamide with DOTA-Bn-SCN (L7 and L8) and DOTAGA (L9). The coupling reaction of DOTA-Bn-SCN was carried out at 40 °C for 4 h. In contrast, for the chelator DOTAGA, the reaction mixture was first sonicated at room temperature for 1 h, resulting in high yields of the coupling product. Two ligands, L10 and L11 (analogs of L1 using a cyclohexyl linker), were also synthesized as shown in Scheme 5. [ka]

[0189] Scheme 5. a Reagents and conditions: (a) (i) trans-4-(Fmoc-aminomethylcyclohexanecarboxylic acid), TSTU, DIPEA, DMF, 1 h at room temperature; (ii) 20% piperidine / DMF; (c) DOTA-GA anhydride, DIPEA, DMSO, 3 h; (c) 50% TFA / CHCl.

[0190] First, the NHS-ester of trans-4-(Fmoc-aminomethylcyclohexanecarboxylic acid) was synthesized in situ and reacted with 1a to give 10. After subsequent removal of the Fmoc and tert-butyl groups and coupling with the corresponding chelators (DOTAGA and DOTA-monoamide), L10 and L11 were obtained in excellent yields. Ligand L12 was synthesized as shown in Scheme 6. Briefly, compound 1a was reacted with intermediate 6, followed by removal of the tert-butyl group from Lys-urea-Glu to give L12 in >80% yield. [ka]

[0191] Scheme 6. (a) 6, DIPEA, DMSO; (b) 50% TFA / CH2Cl2.

[0192] Ligand L13 was synthesized according to Scheme 7, but by replacing the p-bromobenzyl with a p-bromopyridyl group to investigate the effect of the pharmacophore on tumor targeting and drug pharmacokinetics. Ligand L14 was synthesized according to Scheme 7. First, the NHS-ester of p-(iodophenyl)butyric acid (compound 12) was synthesized in quantitative yield, which was then used to prepare the previously reported bifunctional compound 14. 35 to give L14 in excellent success. All newly synthesized ligands were purified by HPLC and lyophilized to give colorless, hygroscopic solid compounds, which were characterized by standard spectroscopic tools, including mass spectrometry and NMR. All ligands were found to be stable for at least 6 months at -20 °C. [ka] Scheme 7. a Reagents and conditions: (a) TSTU, DIEA, DMF; (b) 13, DIPEA, DMSO, 2 h.

[0193] All new ligands were synthesized in high yields by incubation at 70°C for 1 hour. 177 The radiolabeled compounds were radiolabeled with Lu. Subsequent HPLC purification removed unreacted ligand, ensuring radiochemical purity of >99% and specific activity (>37 MBq / nmol) in the highest cases. Furthermore, a rapid and simple microwave-assisted radiolabeling method was developed for L1 and related ligands (including L7, L8, L9, L11, L12, and L13) with excellent yields (>90%) within 5 minutes at 40°C and pH ∼4. 177 Lu labeling agent ( 177We have newly synthesized a number of radiolabeled products using p-iodobenzylurea derivatives (e.g., L4 and L6), long linkers (e.g., L3 and L5), and rigid linkers (e.g., L12) by microwave-assisted methods. 177 The Lu-labeled compound was stable for up to 4 hours at room temperature and 24 hours at 4°C, with no significant radiolysis observed. However, to ensure high stability, L-ascorbic acid was added to the radiolabeled compound in the final formulation used in the in vivo experiments described herein. Cellular uptake and internalization experiments were performed immediately without the addition of L-ascorbic acid.

[0194] 5.3.2 In Vitro Binding and Cellular Uptake. All new ligands have K values ​​in the range of 0.03–8 nM. i The compounds exhibited high binding affinity to PSMA, with values ​​of 0.01 and 0.10 (Table 6). A stable lutetium analog of L1 (Lu-L1) was synthesized, which showed a three-fold improvement in binding affinity over L1 (Table 6). Comparative cellular uptake studies with L1–L14, etc., after 2-hour incubation revealed approximately 100-fold higher uptake in PSMA+ PC3 PIP cells compared with PSMA- PC3 flu cells. Cellular uptake and internalization data for selected compounds are shown in Figure 14 and Table 7. Total uptake and internalization for the compounds gradually increased from 1 hour to 24 hours. 177 Lu-PSMA-I&T and 177 Both Lu-PSMA-617 and Lu-PSMA-617 showed significantly higher total uptake in PSMA(+) PC3 PIP cells (approximately 60% and 40% of the incubated dose, respectively). 177 Lu-L1, 177 Lu-L3 and 177Lu-L5 showed uptake in the 30% range. However, for these compounds, the percent internalization was within the same range: approximately 18-24% at 1 hour and 25-30% at 24 hours. For all compounds, uptake in PSMA(+) PC3 PIP cells could be blocked by excess treatment with a known PSMA inhibitor (ZJ43) (Table 7). 177 Lu-7 and 177 The internalization of Lu-8 was also examined up to 24 hours after incubation. 177 Lu-8 was detected at all time points, and the ligand-free 177 Compared to Lu-7, it showed approximately >1.5-fold higher internalization. Importantly, it possesses the DOTAGA chelator. 177 Lu-9 contains the same chelating agent 177 Lu-PSMA-I&T showed comparable cellular uptake and internalization (approximately 70%) (Table 7). In contrast, when modified with a rigid cyclohexyl linker, the DOTAGA chelator 177 The Lu-10 177 Lu-8 and 177 Compared to Lu-9, it showed lower uptake and internalization. 177 Lu-L11 also contains a cyclohexyl linker, 177 >1.5-fold lower uptake compared to Lu-L1 and 177 It showed lower uptake compared to Lu-10. Furthermore, it has a rigid aromatic linker. 177 Lu-L12 showed lower uptake compared to most drugs from that series. 177 Lu-13 is 177 It showed similar cellular uptake and internalization properties to Lu-1. 177 Lu-14 showed relatively higher uptake in PSMA(+) PC3 PIP cells at 2 hours.

[0195] [Table 6]

[0196] [Table 7]

[0197] [Table 8]

[0198] 5.3.3 Biodistribution Studies. Biodistribution studies were performed in male NOD-SCID mice bearing both PSMA+ PC3 PIP and PSMA-PC3 flu tumors in the upper flank. 177 The tumor uptake and in vivo pharmacokinetics of the Lu compounds were evaluated. A radiotracer dose of ∼1.67 ± 0.2 MBq (∼45 ± 5 μCi) was administered intravenously for all studies presented herein. Based on initial biodistribution studies 24 hours after injection, several ligands were selected for detailed biodistribution studies at longer time points, up to 72–96 hours. Compounds 177 Lu-7 (without Br-benzyl modified urea group) and rigid linker 177 Lu-11 and 177 With the exception of Lu-12, all compounds were highly uptaken into PSMA(+) tumors, exceeding 18% ID / g 24 hours after injection. 177 Lu-PSMA-617, 177 Lu-PSMA-I&T, 177 Lu-1, 177 Lu-3 and 177 The tissue distribution of Lu-5 is shown in Figures 15 and 16. The body weights of the mice and the tumor weights are shown in Table 9.

[0199] [Table 9]

[0200] Three hours after injection, 177Lu-3, 52.6±4.9 %ID / g, and 177 A significantly higher tumor uptake was observed for Lu-5 (56.3±18.3), whereas 177 Tumor uptake in Lu-1 177 Lu-PSMA-617 and 177 The results were comparable to those of Lu-PSMA-I&T. However, after 24 hours, 177 Lu-3 and 177 Lu-5 was rapidly cleared from PSMA+ PC3 PIP tumors due to its uptake. 177 Lu-PSMA-617 and 177 Lu-PSMA-I&T induced tumor uptake comparable to that of Lu-PSMA-I&T. 177 Lu-1 is 177 Lu-PSMA-617 showed significantly lower tumor uptake than Lu-PSMA-617. At 72 hours post-injection, we observed no significant differences in tumor uptake for the compounds. For all compounds, low uptake (<0.3% ID / g) in PC3 flu tumors starting 3 hours post-injection indicates high specificity of the compounds, consistent with the high binding affinity of the ligands.

[0201] The drug was taken up by tumors in a similar manner up to 72 hours after injection, but significant changes were observed in the uptake of the drug into normal tissues. Normal organs that express PSMA (e.g., kidneys, salivary glands, and spleen) 177 The renal uptake was significantly higher than that of Lu-PSMA-I&T. 177 Lu-PSMA-I&T had 93.39±13.35% ID / g at 3 hours after injection, while the remaining drugs from that series showed uptake of <10% ID / g as follows; 177 Lu-PSMA-617 (9.81±6.54% ID / g), 177 Lu-1 (5.17±2.38 %ID / g), 177 Lu-3 (7.49 ± 3.21 %ID / g). 177Lu-PSMA-I&T showed rapid renal clearance of 30.39±12.49 %ID / g at 24 hours, but the drug showed a decrease in renal uptake by approximately 3-fold every 24 hours up to 72 hours. 177 Lu-3 and the remaining compounds had much faster rates of renal clearance (~10-fold clearance), resulting in renal uptake of <0.5 ID / g after 24 hours. 177 With the exception of Lu-PSMA-I&T, uptake in blood and normal tissues, including liver, lung, stomach, pancreas, spleen, fat, adrenal gland, muscle, small intestine, large intestine, bone, and salivary gland, was <0.5 %ID / g at 3 hours post-dose. 177 Despite high uptake in the spleen and salivary glands after 3 hours, Lu-PSMA-I&T was rapidly cleared within 24 hours, demonstrating uptake in these tissues comparable to that shown by other agents (<1% ID / g).

[0202] Selected, 177 p-iodobenzyl analogues of Lu-1 as well as halobenzyl-modified urea-ligands without targeting moieties, 177 Lu-L7 and 177 Lu-L14, as well as bromobenzyl-modified drugs, 177 Lu-L8, 177 Lu-L9 and 177 The tissue biodistribution data for Lu-L10 is shown in Figure 17. 177 Lu-2 is 177 Compared to Lu-L1, it showed significantly higher tumor uptake 3 hours after injection, but tumor uptake and retention was observed for up to 72 hours. 177 Lu-L7 and 177 Lu-L8 was modified with a DOTA-Bn-SCN chelator. 177 Compared to Lu-L7, 177Lu-L8 showed significantly higher tumor uptake and retention at all time points, further highlighting the importance of the bromobenzyl group for tumor retention (55.4 ± 7.2 vs. 25.4 ± 7.2 % ID / g at 2 h; 40.6 ± 7.0 vs. 7.0 ± 1.5 % ID / g at 24 h; 27.0 ± 7.0 vs. 7.0 ± 3.3 % ID / g and 24.9 ± 2.3 vs. 2.3 ± 0.0 % ID / g at 48 h). 177 It showed much faster renal clearance compared to Lu-PSMA-I&T.

[0203] DOTAGA has chelating agents 177 Lu-L9 was detected up to 72 hours after injection. 177 Compared to Lu-PSMA-I&T, Lu-PSMA-I&T showed significantly higher tumor uptake and retention, but also much faster renal clearance (approximately 30-fold higher renal clearance within 24 hours (149.7±32.0% ID / g at 2 hours vs. 5.9±2.7% ID / g at 24 hours)). The rigid cyclohexyl linker did not result in higher cellular uptake compared to L1 ( 177 Lu-L10 and 177 Lu-L11), respectively 177 Lu-L9 and 177 Compared to Lu-L1, drugs with various targeting ligands (2-pyridyl) showed similar or lower uptake. 177 Lu-L13 is 177 It maintained similar tumor uptake to Lu-L1 but was cleared much more rapidly from normal tissues.

[0204] 177 The biodistribution of Lu-14 was consistent with that reported for albumin binders, with highest tumor uptake at 24 hours post-injection and remaining elevated for up to 48 hours. 177Compared to Lu-PSMA-I&T, it showed lower initial renal uptake (49.49 ± 19.55 %ID / g), but only a roughly threefold decrease in activity (17.47 ± 4.18 %ID / g at 48 hours) was observed, compared to a 10-fold decrease in activity (93.39 ± 13.35 %ID / g at 24 hours vs. 9.55 ± 3.85 %ID / g at 48 hours). The drug also showed the highest blood uptake in the series (16.13 ± 2.33 %ID / g at 2 hours post-injection, followed by 5.05 ± 0.05 %ID / g at 2 hours and 2.48 ± 0.44 %ID / g at 48 hours). Both the spleen and salivary glands showed the highest nonspecific uptake from this series.

[0205] 5.3.4 SPECT / CT imaging. 177 Lu-1 and 177 SPECT / CT imaging with Lu-14 was performed to confirm the in vivo pharmacokinetics. As expected from the biodistribution data, 177 SPECT / CT imaging performed 2 to 192 hours after Lu-1 administration confirmed, as expected, high uptake in PSMA+ PC3 PIP tumors (right side) but not in PSMA- PC3 flu tumors (left side). Also consistent with the biodistribution data, the ligand showed very low uptake in the kidney and all normal tissues. The status of PSMA expression in PSMA(+) PC3 PIP tumors was also examined in imaging experiments. As shown in Figure 18B, compared to control tumors (no radioactive injection), 177 A significant decrease in PSMA(+) staining was observed in PC3 PIP tumors from days 1 to 12 after treatment with Lu-1 (37 MBq), partly due to the treatment's effect of downregulating PSMA expression and partly due to the presence of a significant number of PSMA binding sites in the treated tumors. 177 Although low, the relatively higher staining seen after 8 and 12 days may be due to the binding of Lu-1 to the tumor. 177 This coincides with the clearance of Lu-1.

[0206] 111 MBq (3 mCi) 177 Lu-PSMA 617, 177 Lu-2, 177 Lu-4 and 177 A comparison of hematological and blood chemistry parameters in the Lu-6-treated mice group (n=3) after 8 weeks is shown in Table 10.

[0207] [Table 10]

[0208] 5.3.5 Radionuclide therapy in cellular and animal models. 177 Lu-1 and 177 The clonogenic efficiency of PSMA(+) PC3 PIP cells after 48 hours of incubation with Lu-8 is shown in Figure 26. The cell survival fraction was >0.8 for the drug when the cells were incubated for 2 and 24 hours. After 48 hours of incubation, a significant loss of colony survival was observed. 177 Lu-1 and 177 The D0 (37% survival) for Lu-8 was in the same range of 0.3–0.6 μCi / mL.

[0209] NOD / SCID mice (n=10 / group) bearing PSMA+ PC3 PIP tumors were treated with 111 MBq (3 mCi) of the drug ( 177 Lu-PSMA-617, 177 Lu-1, 177 Lu-3 and 177A pilot treatment study was conducted in which a single intravenous dose of 100 mg ... 177 Lu-3 and 177 Only one mouse in the Lu-PSMA-617 treated group, as well as at the high dose 177 Two mice in the Lu-4 group reached tumor volumes >4 mm. After 8 weeks, three mice from each treatment group were used for detailed blood analysis and kidney anatomical examination. Selected metabolic and complete blood count data are provided in Table 3, Figures 24 and 25. All animals in the radiation therapy study had normal creatinine (0.3-0.4 mg / dL) and blood urea nitrogen levels. As shown in Figure 11, 177 Only one mouse in the Lu-PSMA-617-treated group had elevated neutrophil counts (later to 3.5 ± 3.8 K / ml; control, 2.6 K / ml). 177 All three mice treated with Lu-PSMA-617 had significantly lower platelets compared with the control and other treatment groups.

[0210] Pathological examination of H&E staining of an extensive panel of normal tissues revealed only moderate changes in the treatment groups compared with the control group. Significant and relevant changes were primarily identified in the testis and lacrimal gland for all treatment groups (Figure 25). 177 One mouse treated with Lu-5 and 177 In all three mice treated with Lu-PSMA-617, the most significant changes were seen in the testes. Kidney changes were minimal, with only minor tubular changes observed in all treatment groups. The parotid gland (normally adjacent to the extraorbital lacrimal gland) was clearly spared, and Lu-PSMA-617 significantly improved the survival of the testes.177 In addition, the infraorbital lacrimal gland showed mild changes in the 177 Similar changes were particularly evident in the extraorbital lacrimal glands in mice treated with Lu-PSMA-617. 177 Some mice treated with Lu-5 developed thymic lymphomas with a morphology consistent with thymic T-lymphoblastic lymphoma, the expected cause of death in NOD / SCID mice, usually beginning at approximately 6 months of age. T-lymphoblastic lymphomas are promoted by irradiation and several carcinogens and are associated with endogenous retroviral / retroelement interactions in the NOD / SCID mouse strain. 177 Some mice treated with Lu-PSMA-617 developed tumors and metastases in the lungs (<1 mm), as well as tumor cells in blood vessels, body cavities, and serosal surfaces (carcinomatosis), which were evident on blood smears. The cells were up to 25 μm in size, had very large nuclei (in mice), usually with two to three nucleoli, and underwent a high rate of mitosis. Degenerative changes in the testes and lacrimal glands indicated that this animal had received a similar treatment to the other treated mice.

[0211] All animals tested had normal creatinine levels (0.3-0.4 mg / dL) and blood urea nitrogen concentrations.

[0212] 177 Lu-4 and 177 Treatment monitoring for Lu-6 has been completed, 177 Lu-1 and 177 Lu-PSMA-617 was evaluated at a pre-specified endpoint (tumor volume 1800 mm 3 The treatment trial continued until a weight loss of >15% was achieved. 177 Lu-1 and 177 Survival data for Lu-PSMA-617 are shown. 177 The median survival time for Lu-PSMA-617 was 133 days, compared with 177 For Lu-1, it was 234 days. In contrast, this treatment group did not show any changes. Individual tumor volume measurements for each animal are shown in Figure 24. 177Considering that Lu-1 showed a good therapeutic effect, a growth retardation study was performed at increasing doses of 18.5 MBq (0.5 mCi), 37 MBq (1 mCi), and 111 mMBq (3 mCi) (n=5 / group) up to 90 days. As shown in Figure 23, 177 Lu-1 demonstrated significant tumor growth delay in all treatment groups compared with the untreated group. Four of five mice treated with the 0.5 mCi dose achieved a relative tumor volume >5 by week 8 after treatment, whereas only one mouse in the 37 MBq group showed a similar effect. Three mice treated with both the 37 MBq and 111 MBq doses showed complete tumor regression by day 120, demonstrating a therapeutic effect similar to that observed in the initial treatment study.

[0213] Using a PC3 PIP flank tumor model, a dose of 111 MBq was used from 24 hours to 192 hours after treatment. 177 A theranostic (SPECT / CT imaging and treatment) study of Lu-8 (n = 10) was also conducted. SPECT / CT imaging showed 177 A profile very similar to Lu-1 (low normal tissue and kidney uptake) was evident, as expected from the biodistribution data. 177 There was a relatively higher background uptake compared to Lu-1. Three mice (used for SPECT / CT imaging) died unexpectedly, but only two mice showed tumor growth of a relative volume >5 after 8 weeks. The remaining treated mice survived tumor-free for 10 months and were evaluated for toxicity.

[0214] 5.3.6 Discussion A new series of PSMA-based small molecules for the treatment of patients with metastatic prostate cancer 177 The SAR of Lu-labeled theranostic drugs was investigated. 177 Lu has issues with emission characteristics, manufacturability, and radiation safety. 131 I and 90Because it is better than Y, it is the beta-emitting isotope of choice for PSMA-targeted and other cancer therapeutic applications. The primary motivation for this study is the need to understand the molecular and structural origins, i.e., the PSMA-targeting moiety Lys-Glu-urea interaction in the PSMA binding site, which explains the significant PK differences between Type I and Type II. With this understanding, 177 Lu-PSMA-617 and 177 This will allow us to move forward with the search for Type II agents with fewer radiation-related side effects compared to Lu-PSMA I&T. The theranostic agents of the present invention are built on a well-studied, long-linker-based targeting platform. 17, 21, 26, 36-42 From a rigorous synthetic perspective, it was intended to expand the scope of radiotheranostic agents and chemical space for compounds that bind to PSMA. No other synthetic approach as vigorous as this one is known in the art to be directed toward developing low molecular weight PSMA-based radionuclide therapeutics.

[0215] The compounds of the present invention were synthesized using optimized solution-phase chemistry. Therefore, the method of the present invention is easily adaptable to industrial-scale preparation and is expected to be less expensive than solid-phase peptide synthesis. These radiolabeled therapeutic agents were synthesized with as specific radioactivity as possible by separating the free ligand from the radioactive peak. Furthermore, microwave-assisted radiolabeling enabled rapid and high-yield production of radiolabeled compounds at a low temperature of 40°C. Systematic studies of cellular uptake and internalization, followed by tissue biodistribution, revealed several important findings.

[0216] First, the attachment of a p-halobenzyl moiety to the PSMA-targeting Glu-Lys-urea resulted in high tumor uptake and low nonspecific binding to normal tissues. Second, SAR studies demonstrated that the macrocyclic chelator DOTA-Bn-SCN ( 177 Lu-8) and DOTAGA ( 177Lu-9) agents are DOTA-monoamide chelators with three acetylated arms (e.g., 177 These drugs were taken up and retained in the tumor at a higher level than those of the IL-1. 177 It exhibits higher plasma binding compared to Lu-1 and 177 Lu-PSMA I&T showed higher initial renal uptake and much faster renal clearance. Third, compared to rigid linkers, drugs with p-bromobenzyl moieties showed higher tumor uptake and retention with linear linkers (e.g., 177 Lu-1 pair 177 Lu-11, and 177 Lu-9 vs. 177 This observation was also reflected in cellular uptake studies. 177 Lu-10 / 177 The cyclohexyl linker containing Lu-11 exhibited higher intracellular uptake.

[0217] Fourth, as reported by others recently, the addition of an albumin-binding moiety may increase serum half-life. 43, 44 , 177 Lu-14 was retained in the tumor for a longer period. However, as reported elsewhere, these agents were not clinically significant. 177 Lu-PSMA-617 or 177 These drugs are associated with significantly higher renal retention compared to Lu-PSMA-I&T. Given that these drugs exhibited distinct characteristics associated with Type I drugs, 131 I-MIP1095 2, 12, 13 As recognized in 177 Compared to Lu-PSMA-617, much higher uptake in the salivary and lacrimal glands is expected. This autopsy study showed severe abnormalities in normal organs, especially in the lacrimal gland. 177 It has been revealed that the lacrimal gland can be used as a surrogate organ for Lu-PSMA agents. Although most preclinical studies have not reported uptake in the lacrimal gland, high uptake in the lacrimal gland was demonstrated for the albumin binder CTT1403.22 .

[0218] Fifth, renal uptake is low according to self-interference tests. 177 Lu-1 produced a 10-fold increase in renal blockade without significant changes in tumor uptake, while showing high initial renal uptake 177 Lu-9 produced significant tumor uptake blockade and demonstrated similar efficacy, suggesting that amino acid-based nephroprotectants such as D-lysine / polyglutamic acid may be effective. 177 This is an important finding, as it has been shown that the renal uptake of Lu-PSMA I&T could not be improved 45 The results of this study confirmed that uptake in renal proximal tubule cells was due, in part, to PSMA expression. 46 Nephrotoxicity is 177 Although not a major problem with Lu-PSMA radionuclide therapy, this blocking strategy 225 Salivary gland-associated radiation toxicity of PSMA-based alpha particle therapy using Ac-PSMA-617 47 and the long-term nephrotoxicity often obtained with alpha particle-based RPT. 31, 48 It can be useful for:

[0219] 5.4 Materials and Methods Solvents and chemicals purchased from commercial sources were analytical grade or better and were used without further purification. Diisopropylethylamine (DIEA), triethylamine (TEA), lutetium(III) nitrate, N,N,N',N'-tetramethyl-O-(N-succinimidyl)uronium tetrafluoroborate (TSTU), p-aminomethylbenzoic acid, Boc-5-aminovaleric acid, Boc-6-aminohexanoic acid-N-hydroxysuccinimide, and disuccinimidyl suberate were purchased from Sigma-Aldrich. DOTA-tris(t-butyl ester)-monoacid (B270) and DOTA-NHS-ester (B280) were purchased from Macrocyclics, Inc. (Dallas, TX). Carrier-free [ 177Lu]Cl3 (NEZ307000MC) was purchased from PerkinElmer Health Sciences Inc. (Shelton, CT, USA). Analytical thin-layer chromatography (TLC) was performed using Aldrich aluminum-backed 0.2 mm silica gel Z19, 329-1 plates and visualized with ultraviolet light (254 nm), I2, and 1% ninhydrin in ethanol. Flash chromatography was performed using silica gel purchased from Bodman (Aston PA), MP SiliTech 32-63 D 60A. All experiments were performed in duplicate or triplicate to ensure reproducibility. HPLC purification of non-radiolabeled compounds was performed using Phenomenex C 18 Luna 10×250 mm 2 The column was run on an Agilent 1260 Infinity LC system (Santa Clara, CA) and eluted with water (0.1% TFA) (A) and CH3CN (0.1% TFA) (B). A gradient HPLC method was used, including 88 / 22 water / CH3CN for 1-5 min, followed by 88 / 12 water / CH3CN for 0-5 min, and then 88 / 22 water / CH3CN to 44 / 56 water / acetonitrile for 5-25 min, at a flow rate of 8 mL / min. Bruker Ultrashield 登録商標 With a 500 MHz spectrometer 1H NMR spectra were recorded. Chemical shifts (δ) are reported in ppm downfield, referenced to proton resonances due to incomplete deuteration of the NMR solvent. Low-resolution ESI mass spectra were obtained on a Bruker Daltonics Esquire 3000 Plus spectrometer. High-resolution mass spectra were obtained at the University of Notre Dame Mass Spectrometry & Proteomics Facility, Notre Dame, IN, using ESI either by direct injection on a Bruker micrOTOF-II or by LC elution on an ultra-high-pressure Dionex RSLC column using a C18 column coupled to a Bruker micrOTOF-Q II. The compounds Di-tert-butyl(((S)-6-((4-bromobenzyl)amino)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamic acid, 5a, and Di-tert-butyl(((S)-1-(tert-butoxy)-6-((4-iodobenzyl)amino)-1-oxohexan-2-yl)carbamoyl)-L-glutamic acid, 5b, were prepared using the following reported method with slight modifications. 49 Elute with 70-80% MeOH / HO. 18 The crude product was purified using column chromatography to give 0.90 g (62%) of an oil.

[0220] 5.4.1 Analytical data of representative compounds Representative compounds of formula (I) are provided in Chart 1. Chart 1: [ka]

[0221] The following compounds are representative of compounds of formula (I): [ka] [ka] [ka]

[0222] Ligands L1 and L2 were synthesized according to standard synthetic routes as shown in Scheme 2. L2 will be described in detail below.

[0223] (14S,18S)-9-(4-Bromobenzyl)-2,8,16-trioxo-1-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-3,9,15,17-tetraazaicosane-14,18,20-tricarboxylic acid (L1). A mixture of Boc-5-aminovaleric acid (0.087 g, 0.40 mmol), TSTU (0.121 g, 0.40 mmol), and DIPEA (0.103 g, 0.80 mmol) was stirred in DMF (1 mL) at room temperature for 1 h. After dilution with DMF (1 mL), compound 1a (0.264 g, 0.40 mmol) was added dropwise. The reaction mixture was stirred for 4 h, concentrated, and purified by elution with 100% C in 0.1% TFA. 18 Purification by column chromatography gave 0.151 g (44%) of an oily substance as compound 2a. 1H NMR (500 MHz, DMSO-d6) δ ppm 7.95 (s, 1H), 7.40 (d, J = 10 Hz, 1H), 7.34 (d, J= 10 Hz, 1H), 7.24 (s, 1H), 7.04 (d, J= 5 Hz, 1H), 6.97 (d, J = 10 Hz, 1H), 5.48-5.44 (m, 1H), 4.87-4.83 (m, 1H), 4.48-4.36 (m, 2H), 4.26-4.21 (m, 2H), 3.66-3.63 (m, 1H), 3.12-2.95 (m, 4H), 2.90 (s, 1H), 2.81 (s, 1H), 2.73 (s, 2H), 2.33-2.27 (s, 1H), 2.26-2.23 (m, 3H), 2.00-1.98 (m, 1H), 1.77 (m, 1H), 1.66-1.60 (m, 2H), 1.37 (s, 36H), 1.26-1.07 (m, 2H); ESMS m / z: 857.3 (M + H) + A cold solution of 2a (0.145 g, 0.17 mmol) in 50% TFA / CH2Cl2 (2 ml) was added and stirred at room temperature for 2 h. The reaction mixture was concentrated and purified by elution with 40% acetonitrile / water. 18 Purification by column chromatography and lyophilization gave 0.067 g (67%) of a white solid product as compound 3. 1 H NMR (500 MHz, DMSO-d6) δ ppm 7.55 (d, J= 5.0 Hz, 1H), 7.48 (d, J = 5.0 Hz, 1H), 7.19-7.15 (m, 2H), 4.62-4.53 (m, 2H), 4.33-4.27 (m, 2H), 3.40 (s, 1H), 2.98-2.92 (m, 2H), 2.83 (s, 1H), 2.56 (s, 1H), 2.44 (s, 3H), 2.16 (bs, 1H), 1.93-1.84 (m, 2H), 1.74 (s, 2H), 1.67-1.60 (m, 5H), 1.41-1.40 (m, 2H); ESMS m / z: 589.1 (M + H) +A reaction mixture of DOTA-NHS-ester (0.090 g, 0.12 mmol), 3a (0.069 g, 0.08 mmol), and DIPEA (0.102 g, 0.79 mmol) was stirred at room temperature for 3 h. The reaction mixture was concentrated and purified by HPLC to give the desired ligand L1. 1 H NMR (500 MHz, DMSO-d6) δ ppm 12.50 (bs, 5H), 8.38 (bs, 1H), 7.57 (d, J = 5 Hz, 1H), 7.51 (d, J = 5 Hz, 1H), 7.18-7.12 (m, 2H), 6.55 (bs, 1H), 6.37-6.29 (m, 2H), 4.53 (s, 1H), 4.46 (s, 1H), 4.11-4.04 (m, 3H), 3.80 (bs, 4H), 3.25-2.77 (m, 10 H), 2.39-2.37 (m, 2H), 2.27-2.20 (m, 4H), 1.94-1.92 (m, 1H), 1.72-1.63 (m, 2H), 1.65-1.39 (m, 9H), 1.28-1.22 (m, 4H); HRESI-MS: Calcd. for C 40 H 62 BrNO 15 , 973.3513 [M+H] + , Actual value: 973.3542.

[0224] (14S,18S)-9-(4-Iodobenzyl)-2,8,16-trioxo-1-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-3,9,15,17-tetraazaicosane-14,18,20-tricarboxylic acid (L2). Compound L3 was prepared according to the same procedure as in Scheme 3 by using 1b as the starting reactant and 2c and 3c as intermediates. Compound 7c was prepared using the same procedure as described for 7a, but using 5b as the starting material. The crude product was purified using column chromatography eluting with 10% acetone / CHCl to give a colorless oil. 1H NMR (500 MHz, DMSO-d6) δ ppm 7.45 (m, 1H), 7.35 (t, J = 5 Hz, 1H), 7.29 (m, 1H), 7.21 (m, 1H), 7.15 (d, J = 5 Hz, 1H), 5.70 (m, 1H), 4.58-4.47 (m, 1H), 4.35-4.27 (m, 2H), 4.09 (m, 1H), 3.40-3.30 (m, 1H), 3.16-3.08 (m, 2H), 2.80 (s, 2H), 2.41-2.26 (m, 4H), 2.09-2.04 (1H), 1.85-1.80 (m, 1H), 1.75-1.62 (m, 4H), 1.58-1.52 (m, 3H), 1.44 (m, 27H), 1.35 (m, 6H).

[0225] Compound 3c was prepared using the same procedure as described for 3a, starting with 2c. The crude product was purified using HPLC. Spectral data for L2: 1 H NMR (500 MHz, DMSO-d6) δ ppm 8.39 (bs, 1H), 7.38 (t, J = 10 Hz, 1H), 7.31 (m, 1H), 7.21-7.18 (m, 2H), 6.36-6.29 (m, 2H), 4.56-4.50 (m, 2H), 4.11-4.01 (m, 3H), 3.82 (s, 3H), 3.59 (s, 4H), 3.19-3.17 (m, 9H), 3.06 (s, 9H), 2.39 (t, J = 5 Hz, 1H), 2.30-2.23 (m, 3H), 1.91 (m, 1H), 1.73 (m, 1H), 1.63-1.39 (m, 8H), 1.24 (m, 2H); HRESI-MS: Calcd. for C 40 H 61 IN8O 15 , 1020.3301 [M+H] + .

[0226] (14S,18S)-9-(4-Bromobenzyl)-1,8,16-trioxo-1-(4-((2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetamido)methyl)phenyl)-2,9,15,17-tetraazaicosane-14,18,20-tricarboxylic acid (L3). 7b was prepared in a similar manner to 7a, substituting Boc-6-aminohexanoic acid-N-hydroxysuccinimide for Boc-5-aminovaleric acid. Yield 0.115 g (46%) of an oil. 1 H NMR (500 MHz, DMSO-d6) δ ppm 7.41 (d, J= 10 Hz, 1H), 7.35 (d, J = 10 Hz, 1H), 7.04 (d, J = 5 Hz, 1H), 6.97 (d, J = 10 Hz, 1H), 5.47 (d, J = 10 Hz, 1H), 5.10-5.03 (m, 1H), 4.63-4.56 (m, 1H), 4.44 (s, 1H), 4.39 (s, 1H), 4.27-4.22 (m, 2H), 3.29-3.21 (m, 1H), 3.07-3.01 (m, 5H), 2.30-2.19 (m, 4H), 2.03-1.98 (m, 1H), 1.80-1.75 (m, 1H), 1.66-1.58 (m, 7H), 1.49-1.45 (m, 4H), 1.38 (m, 27H), 1.27-1.04 (m, 5H); ESMS m / z: 871.3 (M+H) + Compound 3b was prepared using the same procedure as described for 3a, starting with 2b. The crude product was used directly in the next step without further purification. 1H NMR (500 MHz, DMSO-d6) δ ppm 7.73 (bs, 3H), 7.76 (d, J = 10.0 Hz, 1H), 7.50 (d, J= 10.0 Hz, 1H), 7.18-7.14 (m, 2H), 6.34 (m, 2H), 4.53 (s, 1H), 4.46 (m, 2H), 3.22-3.17 (m, 2H), 2.82-2.73 (m, 2H), 2.51 (s, 1H), 2.37 (s, 1H), 2.25 (m, 2H), 1.93 (s, 1H), 1.74-1.71 (m, 1H), 1.64 (s, 1H), 1.55-1.40 (m, 6H), 1.35-1.33 (m, 1H), 1.25 (s, 4H); ESMS m / z: 603.2 (M + H) + . The reaction mixture of 4 (0.065 g, 0.08 mmol), 3b (0.047 g, 0.08 mmol) and DIPEA (0.101 g, 0.80 mmol) was mixed with DMSO (1 ml) at room temperature for 3 minutes. The reaction mixture was not concentrated and purified by HPLC. 1 H NMR (500 MHz, DMSO-d6) δ ppm 12.65 (brs, 5H), 8.42-8.39 (m, 1H), 7.84-7.81 (m, 2H), 7.56 (d, J = 10.0 Hz, 1H), 7.49 (d, J = 10.0 Hz, 1H), 7.38 (d, J = 5.0 Hz, 2H), 7.17-7.12 (m, 2H), 6.36-6.30 (m, 2H), 4.53 (m, 3H), 4.46 -4.40 (m, 5H), 4.10-3.98 (m, 9H), 3.63 (bs, 5H), 3.26-3.14 (m, 13H), 2.38 (m, 2H), 2.31-2.20 (m, 3H), 1.93-1.92 (m, 1H), 1.72-1.71 (m, 1H), 1.63-1.41 (m, 8H), 1.34 (m, 1H), 1.25 (brs, 3H); HRESI-MS: Calcd. for C 49 H 71 BrN9O 16, 1120.4197 [M+H] + , Actual value: 1120.4200.

[0227] (14S,18S)-9-(4-Iodobenzyl)-1,8,16-trioxo-1-(4-((2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetamido)methyl)phenyl)-2,9,15,17-tetraazaicosane-14,18,20-tricarboxylic acid (L4). Intermediate compound 2d was prepared following the same method as described for 2a by substituting Boc-6-aminohexanoic acid-N-hydroxysuccinimide for Boc-5-aminovaleric acid and substituting 1b for 1a as the reactant. Spectral data for 2d. 1 H NMR (500 MHz, DMSO-d6) δ ppm 7.66 (brs, 1H), 7.61 (d, J = 10 Hz, 1H), 7.55 (d, J = 10 Hz, 1H), 7.15 (brs, 1H), 6.90 (d, J = 5 Hz, 1H), 6.84 (d, J = 10 Hz, 1H), 5.30 (m, 1H), 5.09-5.03 (m, 1H), 4.63-4.58 (m, 1H), 4.43 (s, 1H), 4.39-4.35 (m, 1H), 4.26-4.20 (m, 3H), 3.07-3.00 (m, 3H), 2.30-2.22 ESMS m / z: 917.3 (M+H) + . 3d spectral data. 1H NMR (500 MHz, DMSO-d6) δ ppm 7.87 (d, J = 10 Hz, 1H), 7.72 (d, J = 10 Hz, 1H), 7.66 (d, J = 10 Hz, 1H), 7.30 (d, J = 10 Hz, 1H), 7.04-6.99 (m, 1H), 4.59-4.51 (m, 1H), 4.32-4.25 (m, 3H), 3.69-3.66 (m, 1H), 3.38-3.35 (m, 1H), 3.09 (m, 1H), 2.96-2.89 (m, 1H), 2.51-2.45 (m, 1H), 2.43-2.39 (m, 3H), 2.17-2.14 (m, 1H), 1.91-1.72 (m, 2H), 1.71-1.62 (m, 6H), 1.45-1.28 (m, 8H); ESMS m / z: 649.2 (M + H) + . L4のスペクトル·データ. 1 H NMR (500 MHz, DMSO-d6) δ ppm 9.16 (bs, 4H), 8.41 (m, 4H), 7.82 (t, J = 10.0 Hz, 2H), 7.72 (d, J = 10.0 Hz, 1H), 7.66 (d, J = 10.0 Hz, 1H), 7.37 (d, J = 10.0 Hz, 2H), 7.01 (d, J = 5.0 Hz, 2H), 6.37-6.30 (m, 2H), 4.44-4.39 (m, 8H), 4.11-3.97 (m, 9H), 3.66-3.61 (m, 5H), 3.16-3.08 (m, 13H), 2.30 (m, 1H), 2.29-2.20 (m, 2H), 1.94-1.92 (m, 1H), 1.71 (m, 1H), 1.66-1.64 (m, 2H), 1.55-1.47 (m, 8H); HRESI-MS: Calcd. for C 49 H 71 IN9O 16 , 1168.4058 [M+H] + , measured value: 1168.4045.

[0228] (21S,25S)-16-(4-Bromobenzyl)-1,8,15,23-tetraoxo-1-(4-((2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetamido)methyl)phenyl)-2,7,16,22,24-pentaazaheptacosane-21,25,27-tricarboxylic acid (L5). Ligand L5 was prepared according to a multistep organic synthesis method, as shown in Scheme 4. A solution of 1a (0.190 g, 0.29 mmol), EtN (0.029 g, 0.29 mmol), and DMF (1 mL) was added dropwise to a stirred solution of disuccinimidyl suberate (0.223 g, 0.61 mmol) in DMF (1 mL). The reaction mixture was stirred overnight, concentrated, and purified by flash column chromatography eluting with 30% acetonitrile / CHCl to give 0.120 g (46%) of an oil. ESMS m / z: 911.3 (M + H). + Compound 8 was prepared using the same method as described for 10. The crude product was purified by column chromatography eluting with 40–60% acetonitrile / CHCl to give 0.090 g (40%) of an oil. 1H NMR (500 MHz, CDCDMSO-d6) δ ppm 7.70 (brs, 1H), 7.62 (d, J = 5.0 Hz, 1H), 7.55 (d, J= 10.0 Hz, 1H), 6.91 (d, J = 5.0 Hz, 1H), 6.85 (d, J = 5.0 Hz, 1H), 5.41 (d, J = 5.0 Hz, 1H), 5.33 (d, J = 10.0 Hz, 1H), 5.09-5.02 (m, 1H), 4.45-4.35 (m, 2H), 4.25-4.19 (m, 3H), 3.41-3.34 (m, 1H), 3.12-3.05 (m, 2H), 2.57-2.50 (m, 2H), 2.31-2.18 (m, 5H), 1.97 (m, 1H), 1.86-1.77 (m, 3H), 1.71-1.63 (m, 5H), 1.57-1.46 (m, 5H), 1.37 (m, 27 H); ESMS m / z: 957.2 (M + H) + Compound 12 was prepared using the same method as described for 3a. The crude product was purified by C-18 column chromatography eluting with 40-50% acetonitrile / water to give 0.057 g (58%) of product. ESMS m / z: 743.2 (M + H). + Compound L6 was prepared using the same procedure as described for L1, starting with compounds 1a and 12. The reaction mixture was concentrated and purified by HPLC. 1H NMR (500 MHz, DMSO-d6) δ ppm 8.97 (brs, 1H), 8.43 (t, J = 5.0 Hz, 1H), 7.83 (d, J= 10.0 Hz, 2H), 7.78-7.73 (m, 1H), 7.56 (d, J= 10.0 Hz, 1H), 7.50 (d, J = 10.0 Hz, 2H), 7.38 (d, J = 10.0 Hz, 2H), 7.17-7.14 (m, 2H), 6.36-6.30 (m, 2H), 4.52-4.39 (m, 8H), 4.09-4.00 (m, 6H), 3.65-3.60 (m, 5H), 3.26-3.08 (m, 11H), 3.07-3.04 (m, 3H), 2.36-2.33 (m, 1H), 2.27-2.22 (m, 2H), 2.06-2.01 (m, 2H), 1.93-1.92 (m, 1H), 1.73-1.64 (m, 2H), 1.51-1.43 (m, 11H), 1.26-1.19 (m, 6H); HRESI-MS: Calcd. for C 55 H 82 BrN 10 O 17 , 1233.5037 [M+H] + , Actual value: 1233.5029.

[0229] (21S,25S)-16-(4-Iodobenzyl)-1,8,15,23-tetraoxo-1(4-((2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetamido)methyl)phenyl)-2,7,16,22,24-pentaazaheptacosane-21,25,27-tricarboxylic acid (L6). Compound L7 was prepared using the same method as described for L5. The crude product was purified by HPLC. 1H NMR (500 MHz, DMSO-d6) δ ppm 8.97 (brs, 1H), 8.43 (t, J = 5.0 Hz, 1H), 7.83 (d, J= 10.0 Hz, 2H), 7.78-7.72 (m, 2H), 7.67 (d, J= 5, 1H), 7.55 (bs, 1H), 7.39 (d, J = 10.0 Hz, 2H), 7.03-6.99 (m, 2H), 6.36-6.30 (m, 2H), 4.52-4.39 (m, 4H), 4.11-4.00 (m, 6H), 3.64 (m, 4H), 3.26-3.04 (m, 14H), 2.35-2.33 (m, 1H), 2.27-2.22 (m, 3H), 2.20-2.01 (m, 2H), 1.93-1.92 (m, 1H), 1.73-1.62 (m, 2H), 1.51-1.43 (m, 11H), 1.26-1.19 (m, 6H); HRESI-MS: Calcd. for C 55 H 82 IN 10 O 17 , 1281.4899 [M+H] + , Actual value: 1281.4889.

[0230] (13S,17S)-8-(4-Bromobenzyl)-7,15-dioxo-1-((4-((1,4,7,10-tetrakis(carboxymethyl)-1,4,7,10-tetraazacyclododecan-2-yl)methyl)phenyl)amino)-1-thioxo-2,8,14,16-tetraazanonadecane-13,17,19-tricarboxylic acid (L8). p-SCN-Bn-DOTA (12.2 mg, 17.7 μmol) was added to a stirred solution of 5a (12.2 mg) and DIPEA (15.2 μL, 87.0 μmol) in DMSO (130 μL) equilibrated to 40 °C. The reaction mixture was stirred at 40 °C for 4 h and stored at 4 °C overnight. The reaction mixture was purified by reverse-phase HPLC (hold 20% ACN for 5 min, then 20-40% for 19 min). tApproximately 12 min. The purified fractions were combined, reduced in volume by rotary evaporation, and then lyophilized. ESI-MS: 1138.37 [M+H] + Found: 1138.5. Compound 1 was further purified by HPLC using a gradient method. The HPLC method included a mobile phase of 88% water (with 0.1% TFA) and 22% CHCN (0.1% TFA) for 1-5 min, followed by 88% water (with 0.1% TFA) and 12% CHCN (0.1% TFA) for 0-5 min, and a gradient from 88% water to 44% water and 12% acetonitrile to 56% acetonitrile for 5-25 min, at a flow rate of 8 mL / min.

[0231] (13S,17S)-7,15-Dioxo-1-((4-((1,4,7,10-tetrakis(carboxymethyl)-1,4,7,10-tetraazacyclododecan-2-yl)methyl)phenyl)amino)-1-thioxo-2,8,14,16-tetraazanonadecane-13,17,19-tricarboxylic acid. (L7) p-SCN-Bn-DOTA (8.0 mg, 11.6 umol) was added to a stirred solution of 5a and DIPEA (10.1 uL, 58 umol) in DMSO (150 uL) equilibrated to 40 °C. The reaction mixture was stirred at 40 °C for 4 h, then cooled to room temperature, diluted with water, and finally purified by reverse-phase HPLC (12% ACN hold for 5 min, then 12–32% for 20 min). Rt: approx. 12 min. The purified fractions were combined, evaporated to a reduced volume, and then lyophilized. M / Z calculated, 970.41. M / Z observed, 970.4. ESI-MS: 970.05 [M+H] + , Actual value: 970.1.

[0232] (3S,7S)-12-(4-Bromobenzyl)-5,13,19-trioxo-22-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-4,6,12,18-tetraazadocosane-1,3,7,22-tetracarboxylic acid (L9). A suspension of DOTA-GA anhydride (0.087 g, 0.19 mmol), 8a (0.075 g, 0.13 mmol), and DIPEA (0.066 g, 0.51 mmol) was sonicated for 1 h at room temperature and stirred for an additional 2 h at room temperature. The reaction mixture was concentrated and purified by HPLC. ESMS m / z: 1047.2 (M + H). + . 1 H NMR (500 MHz, DMSO-d6) δ ppm 12.66 (bs, 4H), 7.86-7.82 (m, 1H), 7.56 (d, J = 10 Hz, 1H), 7.50 (d, J = 10 Hz, 1H), 7.15 (m, 2H), 6.34-6.29 (m, 2H), 4.52 (s, 1H), 4.45 (s, 2H), 4.12-4.04 (m, 5H), 3.50 (m, 4H), 3.29 (m, 3H), 3.19 (m, 3H), 3.07-2.90 (m, 8H), 2.38 (m, 3H), 2.25 (m, 3H), 1.94-1.90 (m, 3H), 1.71 (m, 2H), 1.53-1.43 (m, 6H), 1.42 (m, 1H), 1.41-1.23 (m, 3H); ESMS m / z: 1047.2 (M + H) + HRESI-MS: Calcd. for C 43 H 65 BrNO 17, 1047.3714 [M+H] + , Actual value: 1045.3705.

[0233] (((1S)-5-((1R,4r)-N-(4-Bromobenzyl)-4-((4-carboxy-4-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)butanamido)methyl)cyclohexane-1-carboxamido)-1-carboxypentyl)carbamoyl)-L-glutamic acid (L11) (VK03-51). A mixture of trans-4-(Fmoc-aminomethylcyclohexanecarboxylic acid) (0.069 g, 0.18 mmol), TSTU (0.055 g, 0.18 mmol), and DIPEA (0.064 g, 0.50 mmol) in DMF (1 mL) was stirred at room temperature for 1 hour. After dilution with DMF (1 mL), compound 5a (0.110 g, 0.17 mmol) was added dropwise. The reaction mixture was stirred for 4 h, concentrated and purified by silica gel chromatography eluting with 2% MeOH / CH2Cl2 to give 0.90 g (52.94%) of an oily product. 1 H NMR (500 MHz, CDCl3) δ ppm 8.02 (s, 1H), 7.78 (d, J = 5.0 Hz, 3H), 7.60 (d, J = 10.0 Hz, 3H), 7.41 (t, J = 10.0 Hz, 4H), 7.33 (d, J = 10.0 Hz, 3H), 7.08 (m, 1H), 4.79 (m, 1H), 4.46 (m, 3H), 4.22 (m, 3H), 3.15 (m, 1H), 3.08 (m, 2H), 2.81 (s, 1H), 2.59 (t, J = 10.0 Hz, 1H), 2.32 (m, 1H), 2.20 (s, 2H), 2.08 (m, 1H), 1.85 (m, 4H), 1.60-1.44 (m, 27H), 1.26 (s, 1H), 1.05-0.88 (m, 3H). The oily product was then dissolved in 20% piperidine / DMF and stirred at room temperature under N for 3 hours. The reaction mixture was concentrated and chromatographed using 10% MeOH / CHCl as the eluent to give 0.030 g of product (yield=42.8%). ESMS m / z: 795.2 (M + H). +The product (0.030 g, 0.04 mmol) was added to a suspension of DOTA-GA anhydride (0.028 g, 0.05 mmol in 1 ml) and DIPEA (0.024 g, 0.18 mmol), sonicated at room temperature for 1 h, and then stirred at room temperature for 2 h. The reaction mixture was concentrated and purified on a Sep-Pak column using 70-80% ACN / HO as the eluent. ESMS m / z: 1253.4 (M + H). + .

[0234] (((S)-5-(N-(4-bromobenzyl)-4-((2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetamido)methyl)benzamido)-1-carboxypentyl)carbamoyl)-L-glutamic acid (L12). L1 was synthesized according to a multi-step organic synthesis as described in Scheme 2. To a solution of DOTA-tBu-ester, 1 (0.4 g, 0.5 mmol in 7 ml of DMF) was added p-aminomethylbenzoic acid (0.076 g, 0.5 mmol), and the solution was stirred at room temperature overnight. The resulting solution was evaporated under high vacuum. The solid residue was dissolved in 20 / 80 acetonitrile / water and C 18 Purification was carried out on a SepPak column. The product was eluted with 60 / 40, 50 / 50 and 40 / 60 acetonitrile / water fractions. The fractions were combined, evaporated and then lyophilized to give a colorless solid. Yield: 0.36 g (80%). ESMS m / z: 706.5 (M + H). + To a solution of compound 2 (0.2 g, 0.29 mmol in DMF) was added TSTU (0.1 g, 0.34 mmol) and DMAP (0.17 g, 0.1 mmol) and stirred at room temperature overnight. The solution was evaporated under high vacuum. The solid residue was dissolved in 20 / 80 acetonitrile / water and HCl. 18 Purification was performed on a SepPak column. The product, compound 3, was eluted with 50 / 50 and 40 / 60 acetonitrile / water fractions and lyophilized to give a colorless solid. Yield: 0.23 g, 90%. ESMS m / z: 802.96 (M + H). +Compound 3 (0.2 g, 0.23 mmol) was dissolved in ice-cold TFA / CH2Cl2 (1:1) and stirred overnight for 18 h. The reaction mixture was concentrated and purified by elution with 50–60% acetonitrile / water. 18 After purification by column chromatography and lyophilization, 4 was obtained in good yield (0.16 g, ca. 60%). 1 H NMR (500 MHz, DMSO-d6) δ ppm 9.03 (bs, 1H), 8.09 (d, J = 10.0 Hz, 2H), 7.93 (d, J= 10.0 Hz, 1H), 7.59 (d, J = 5.0 Hz, 3H), 7.43 (d, J = 10.0 ESMS m / z: 635.3 (M+H) + Compound 6 was prepared by dissolving 4 (0.200 g, 0.36 mmol) and 5a in DMSO (1 mL). 49 (0.050 g, 0.16 mmol) followed by DIPEA (0.406 g, 3.65 mmol) and the reaction mixture was stirred at room temperature overnight. The crude product was concentrated and used without further purification. A cold solution of 6 (0.050 g, 0.04 mmol) in 50% TFA / CHCl (2 ml) was added to 6 (0.050 g, 0.04 mmol) and stirred at room temperature for 2 h. The reaction mixture was concentrated and purified by HPLC to give the final product L1 in good yield. 1H NMR (500 MHz, DMSO-d6) δ ppm 12.80 (brs, 3H), 8.94 (s, 1H), 7.56 (m, 3H), 7.38-7.34 (m, 4H), 7.15 (m, 1H), 6.35 (d, J = 10 Hz, 2H), 4.64 (m, 4H), 4.43-4.39 (m, 5H), 4.13-3.99 (m, 7H), 3.63 (bs, 4H), 3.14 (m, 9H), 2.37-2.18 (m, 2H), 1.95-1.89 (m, 1H), 1.74-1.68 (m, 1H), 1.55-1.49 (m, 3H), 1.39-1.30 (m, 2H), 1.05 (m, 1H); HRESI-MS: Calcd. for C 43 H 60 BrNO 15 , 1007.3356 [M+H] + , Actual value: 1007.3367.

[0235] (14S,18S)-9-((6-bromopyridin-3-yl)methyl)-2,8,16-trioxo-1-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-3,9,15,17-tetraazaicosane-14,18,20-tricarboxylic acid (L13) (VK02-112). 1 H NMR (500 MHz, DMSO-d6) δ ppm 7.21 (m, 1H), 7.13 (m, 2H), 6.12 (bs, 1H), 5.90 (bs, 1H), 4.29 (s, 1H), 4.19-4.15 (m, 2H), 3.26 (m, 1H), 2.93 (m, 2H), 2.34-2.23 (m, 3H), 2.07-2.02 (m, 1H), 1.82-1.76 (m, 1H), 1.70-1.64 (m, 3H), 1.56-1.53 ​​(m, 1H), 1.43-1.37 (m, 27H), 1.27 (d, J = 5 Hz, 2H); ESMS m / z: 657.2 (M + H) +The product was purified using 70-90% ACN / H2O on a C-18 Sep-Pak column. 1 H NMR (500 MHz, DMSO-d6) δ ppm 8.25 (s, 1H), 7.49 (m, 1H), 7.43 (m, 1H), 5.95 (m, 1H), 5.36 (m, 1H), 4.80 (s, 1H), 4.51 (m, 2H), 4.36-4.33 (m, 3H), 3.24-3.09 (m, 5H), 2.40-2.29 (m, 6H), 2.10-2.05 (m, 2H), 1.85-1.76 (m, 3H), 1.72-1.66 (m, 3H), 1.60-1.36 (m, 36H); ESMS m / z: 856.3 (M + H) + The product was purified on silica gel using 3% MeOH / CH2Cl2 as the eluent. The product, 17.4% yield, was purified on a C-18 Sep-Pak column using 50-60% ACN / H2O. The yield was 82.6%. 1 H NMR (500 MHz, DMSO-d6) δ ppm 8.27 (s, 1H), 7.80 (m, 1H), 7.26-7.13 (m, 1H), 6.41-6.36 (m, 1H), 4.48 (dd, J = 2.0, 1.5 Hz, 1H), 4.11 (m, 2H), 3.23 (m, 1H), 2.79 (bs, 2H), 2.42 (s, 1H), 2.33-2.21 (m, 3H), 1.94 (m, 1H), 1.75-1.65 (m, 2H), 1.57-1.42 (m, 6H), 1.28-1.23 (m, 3H), 1.10 (dd, J = 5, 2.5, 1.5 Hz, 1H); ESMS m / z: 588.2 (M + H) + .

[0236] (3S,7S,26S,29R)-38-(4-iodophenyl)-5,13,20,28,35-pentaoxo-29-(2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetamide)-4,6,12,21,27,34-hexaazaoctatriacontane-1,3,7,26-tetracarboxylic acid (L14) (SR-IX-14 A).

[0237] 5.4.2 Radiolabeling and Serum Stability. For each radiolabeling reaction, 1 mCi of IgG was measured in 200 mM NaOAc (pH 4-5). 177 Approximately 1 nmol of radioligand per Lu was heated in a microwave at 95°C and 40 watts of power for 5 minutes. The reaction solution was diluted with 300 μL of water. Complex formation was monitored by injecting 20-40 μL aliquots of the solution onto an HPLC system. The radiolabeled product [ 177 [Lu]X was obtained in approximately 98% radiochemical yield with a radiochemical purity of >99%. All radiolabeled compounds were radiolabeled using Phenomenex C 18 Luna 10×250 mm 2 Purification was performed using a column and an Agilent Prostar System (Palo Alto, CA) equipped with a Varian ProStar 325 UV-Vis variable wavelength detector and a Bioscan (Poway, CA) flow-counting inline radioactivity detector, all controlled by Galaxie software. The flow rate was 1 mL / min using water (0.1% TFA) (A) and CH3CN (0.1% TFA) (B) as eluents. To ensure uniform purity, isocratic solutions were used to separate excess ligand from the radiolabeled compound. Specific radioactivity was calculated during preparative HPLC purification as the ratio of the radioactivity eluting at the product retention time to the mass corresponding to the area under the UV absorbance curve. The purity of the test compound was >95% as determined by analytical HPLC using absorbance at 254 nm.

[0238] The HPLC method and chromatogram of the radiolabeled compound were obtained (data not shown). The specific activity of the compound was >37 MBq / nmol (n >12). The acidic eluate was neutralized with 50 μL of 1 M Na2CO3 solution, and the volume of the eluate was reduced by drying under vacuum. The solid residue was diluted with saline and 2 μL of ascorbic acid (200 mg / mL) to the desired radioactivity concentration for all biological studies, including biodistribution, imaging, and therapeutic studies. The radiolabeled yield was also assessed using silica gel instant TLC (ITLC) with 10 mM diethylenetriaminepentaacetic acid (DTPA) as the mobile phase. Three microliters of the diluted sample was spotted onto an ITLC silica gel strip and developed in the chromatography chamber. After migration to the solvent front was complete, the ITLC sample strip was dried, cut in half, and counted in a Wallac Wizard γ counter (PerkinElmer, Boston, MA) to determine the radiolabeled yield. Radiochemical purity was assessed by high performance liquid chromatography (HPLC) analysis.

[0239] 5.4.3 Cell Lines. Sublines of the androgen-independent PC3 human prostate cancer cell line, derived from an advanced androgen-independent bone metastasis, were used. These sublines were modified to express high levels of PSMA [PSMA-positive (+) PC3 PIP] or lack the target [PSMA-negative (-) PC3 flu]. They were kindly provided by Dr. Warren Heston (Cleveland Clinic). Cells were grown in RPMI 1640 medium (Corning Cellgro, Manassas, VA) containing 10% fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO) and 1% penicillin-streptomycin (Corning Cellgro, Manassas, VA). PSMA+ PC3 PIP cells were grown in the presence of 20 μg / mL puromycin to maintain PSMA expression. All cell cultures were maintained in a humidified incubator at 37.0°C in an atmosphere containing 5% carbon dioxide (CO2).

[0240] 5.4.4 Cellular Uptake and Internalization Measurements. Cellular uptake studies were performed as previously reported. 50One million cells were incubated with each radiolabeled agent at 37 kBq / mL (1 μCi / mL) in growth medium in six-well plates. To measure specific cellular uptake, cells were pre-blocked with ZJ43 at a final concentration of 10 μM. Cell uptake was terminated by washing with 1 mL of ice-cold PBS. After 20 and 60 min of incubation at 37°C, cells were washed with binding buffer and trypsinized using non-enzymatic buffer, and the activity relative to the cells was measured using a gamma spectrometer (1282 Compugamma CS; Pharmacia / LKB Nuclear, Inc.). For internalization assays, cells were detached using non-enzymatic buffer, and aliquots of one million cells per tube were incubated with each radiolabeled agent at 37 kBq (1 μCi) per milliliter for 1, 2, 4, and 24 h at 37°C. Because receptor endocytosis was considered minimal at 4°C, internalization assays were performed using only cells incubated at 37°C. At 1, 2, 4, and 24 hours, the medium was removed, and the cells were washed once with binding buffer, followed by a mild acidic buffer (50 mM glycine, 150 mM NaCl [pH 3.0]) at 4°C for 5 minutes. The acidic buffer was then withdrawn, and the cells were washed twice with binding buffer. The pooled washes (containing surface-bound ATP) were used to determine the amount of ATP. 177 Lu-labeled reagent) and cell pellet (internalized 177 The cells (containing Lu-labeling agent) were counted along with standards in an automated gamma counter. All radioactivity values ​​were converted to percentage of the incubated dose per million cells (%ID). Experiments were performed in triplicate and repeated three times. Data were fitted according to linear regression analysis using PRIZM software.

[0241] 5.4.5 Biodistribution. Mice bearing PSMA(+)PC3 PIP and PSMA(-)PC3 flu tumor xenografts were administered 1.11–1.85 MBq (30–50 μCi) of 150 μL of saline (n = 4). 177Lu-X was injected via the tail vein. At 10, 30, 60, and 120 minutes after injection, mice were sacrificed by cervical dislocation, and their blood was immediately collected by cardiac puncture. Heart, lung, liver, stomach, pancreas, spleen, adipose tissue, kidney, muscle, small and large intestine, bladder, PSMA(+)PC3 PIP, and PSMA(-)PC3 flu tumors were collected. Each organ was weighed, and tissue radioactivity was measured using an automated gamma counter (1282 Compugamma CS, Pharmacia / LKB Nuclear, Inc., Mt. Waverly, Victoria, Australia). The percentage of injected dose per gram of tissue (% ID / g) was calculated by comparing with a standard dilution sample of the initial dose. All measurements were corrected for decay.

[0242] 5.4.6 In vivo blood plasma protein binding and metabolism in mice (n=2 / group) 177 Lu-1 and 177 Lu-9 radiotracer (3.7 mBq in 150 μL saline) was administered via intravenous injection into the tail vein. Mice were sacrificed 2 hours after administration. Blood was collected in heparinized tubes and centrifuged (5 min, 1700 g) to isolate plasma. Plasma samples (50 μL) were transferred to an ultrafiltration device [Centrifree ultrafiltration device (Millipore Sigma, USA)] and centrifuged to separate proteins. Samples of the filtrate and protein fraction were measured using a gamma counter. Furthermore, ITLC was performed to evaluate the radiotracer stability in the aqueous blood fraction.

[0243] 5.4.7 SPECT / CT imaging of small animals. 177 Lu-1 and 177Lu-14 was imaged using the same model (n = 2) bearing both PSMA(+) PC3 PIP and PSMA(-) PC3 flu tumor xenografts used in the biodistribution study. SPECT-CT imaging for radiotherapy was performed using mice bearing only PSMA(+) PC3 PIP tumors. Mice were anesthetized with 1% isoflurane gas (in oxygen) flowing at 0.6 L / min before and during radiopharmaceutical injection.

[0244] Mice received approximately 37 MBq (1 mCi) or 111 MBq (3 mCi, for therapeutic studies) of 150 μL of saline (pH 7). 177 Lu-X was injected via the tail vein. After 2 hours of radiopharmaceutical uptake, the anesthetic flow rate was increased to 0.8 L / min, while the anesthetized mouse was placed on the scanner gantry and secured with medical tape. The mouse was covered with several layers of Chux disposable padding and illuminated with a dissecting lamp during scanning to maintain its body temperature. SPECT imaging was performed using a 1.0 mm aperture single-pinhole median-energy (PHME) collimator with 64 projection angles, rotated stepwise to a full 360°, in 45 s increments. The radius of rotation was set to 6.5 cm, providing a 7.5 cm field of view covering the mouse's body from head to bladder. Prior to scintigraphy, 14 CT scans were performed for both anatomical registration and attenuation correction. A total of 512 projections were acquired in continuous rotation mode for 2 minutes, covering a full 360° of rotation. Data were reconstructed and fused using commercially available software obtained from the vendor (GammaMedica). Data were analyzed using AMIDE software.

[0245] 5.4.8 Radiopharmaceutical treatment of tumors. 177 Lu-L1, 177 Lu-L3, 177 Lu-L5 and 177To investigate the therapeutic effect of a single intravenous administration of Lu-PSMA-617 compared with saline, an antitumor efficacy study was conducted using a PC3 PIP flank tumor growth retardation model. This was a head-to-head comparison study, and the same batch of tumor-bearing mice and radioactivity were used. 14–18 days after xenograft inoculation (all mice were approximately 60–85 mm ), tumors were observed. 3 The study was initiated when the mice had a tumor volume of 1800 mm or greater (Table 8). All surviving mice were monitored for 8 weeks to monitor tumor growth delay. Mouse weight and tumor volume were monitored every 3 days throughout the experiment. Endpoint criteria defined by the ACUC organization were weight loss >= 15% and tumor volume > 1800 mm. 3 After 8 weeks, three mice from each group underwent detailed necropsy and blood analysis. 177 Lu-1 and 177 The group of mice treated with Lu-PSMA-617 continued to be monitored until the end point. These definitions were also used for Kaplan-Meier analysis. The probability of reaching five times the initial tumor volume was determined using Kaplan-Meier curves and compared using the log-rank test. V = width 2 The formula: × length / 2 was used to calculate tumor volume. 177 Toxicity in Lu-treated and 111 MBq-treated healthy CD-1 (n = 3) mice was assessed by pathological examination at the Johns Hopkins Pathology Core Facility, which included a serum metabolic panel, blood counts, and a complete necropsy including detailed histopathology of the kidneys, salivary glands, and lacrimal glands.

[0246] 5.4.9 Data Analysis. Data are expressed as mean ± standard deviation (SD). Statistical significance was determined using Prism software (GraphPAD, San Diego, California). Statistical significance was calculated using a paired t-test. A P value <0.05 was considered significant.

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[0248] [[ID=I]] References All publications, patent applications, patents, and other references mentioned herein are indicative of the level of skill of those skilled in the art to which the subject matter of this invention pertains. All publications, patent applications, patents, and other references (e.g., websites, databases, etc.) mentioned herein are incorporated by reference in their entirety to the same extent as if each individual publication, patent application, patent, and other reference was specifically and individually indicated to be incorporated by reference. Although numerous patent applications, patents, and other references are referenced herein, it is understood that such reference does not constitute an admission that any of these documents form part of the common general knowledge in the art. In the event of a conflict between this specification and any incorporated reference, the specification (including any amendments thereto, which may be based on the incorporated reference) shall control. Standard, art-accepted meanings of terms are used herein unless otherwise specified. Standard abbreviations for various terms are used herein.

[0249] International PCT Patent Application No. PCT / US2008 / 007947 to Pomper, MG, Ray, S., Mease, RC, Foss, C. for Labeled inhibitors of prostate specific membrane antigen (PSMA), biological evaluation, and use as imaging agents, published 2008 / 12 / 31 (WO 2009 / 002529 A2); International PCT Patent Application No. PCT / US2008 / 013158 to Chandran S.S., Ray S., Denmeade S.R., Pomper M.G., Mease R.C. for Prostate specific membrane antigen targeted nanoparticles for therapy of prostate cancer, published 2009 / 06 / 04 (WO 2009070302 A1); International PCT Patent Application Publication No. PCT / US2010 / 028020 to Pomper M.G., Mease R.C.; Ray S., Chen Y. for PSMA-targeting compounds and uses thereof, published 2010 / 09 / 23 (WO 2010108125 A2); Banerjee, S. R., Foss, C. A., Pullambhatla, M., Wang, Y., Srinivasan, S., Hobbs, R. F., Baidoo, K. E., Brechbiel, M. W., Nimmagadda, S., Mease, R. C., Sgouros, G., and Pomper, M. G. (2015) Preclinical evaluation of 86Y-labeled inhibitors of prostate-specific membrane antigen for dosimetry estimates. Journal of nuclear medicine 56, 628-34; Banerjee, S. R., Pullambhatla, M., Byun, Y., Nimmagadda, S., Green, G., Fox, J. J., Horti, A., Mease, R. C., and Pomper, M. G. (2010) 68Ga-labeled inhibitors of prostate-specific membrane antigen (PSMA) for imaging prostate cancer. J Med Chem 53, 5333-5341; Banerjee, S. R., Pullambhatla, M., Byun, Y., Nimmagadda, S., Foss, C. A., Green, G., Fox, J. J., Lupold, S. E., Mease, R. C., and Pomper, M. G. (2011) Sequential SPECT and optical imaging of experimental models of prostate cancer with a dual modality inhibitor of the prostate-specific membrane antigen. Angewandte Chemie International Edition 50, 9167-9170; Banerjee, S. R., Pullambhatla, M., Shallal, H., Lisok, A., Mease, R. C., and Pomper, M. G. (2011) A modular strategy to prepare multivalent inhibitors of prostate-specific membrane antigen (PSMA). Oncotarget 2, 1244-1253; Ray Banerjee, S., Pullambhatla, M., Foss, C. A., Falk, A., Byun, Y., Nimmagadda, S., Mease, R. C., and Pomper, M. G. (2013) Effect of chelators on the pharmacokinetics of (99m)Tc-labeled imaging agents for the prostate-specific membrane antigen (PSMA). J Med Chem 56, 6108-6121; Banerjee, S. R., Pullambhatla, M., Foss, C. A., Nimmagadda, S., Ferdani, R., Anderson, C. J., Mease, R. C., and Pomper, M. G. (2014) (6)(4)Cu-labeled inhibitors of prostate-specific membrane antigen for PET imaging of prostate cancer. J Med Chem 57, 2657-2669; Ray Banerjee, S., Chen, Z., Pullambhatla, M., Lisok, A., Chen, J., Mease, R. C., and Pomper, M. G. (2016) Preclinical Comparative Study of (68)Ga-Labeled DOTA, NOTA, and HBED-CC Chelated Radiotracers for Targeting PSMA. Bioconjug Chem 27, 1447-1455; Benesova, M., Schafer, M., Bauder-Wust, U., Afshar-Oromieh, A., Kratochwil, C., Mier, W., Haberkorn, U., Kopka, K., and Eder, M. (2015) Preclinical Evaluation of a Tailor-Made DOTA-Conjugated PSMA Inhibitor with Optimized Linker Moiety for Imaging and Endoradiotherapy of Prostate Cancer. Journal of nuclear medicine 56, 914-20; Tykvart, J., Schimer, J., Jancarik, A., Barinkova, J., Navratil, V., Starkova, J., Sramkova, K., Konvalinka, J., Majer, P., and Sacha, P. (2015) Design of Highly Potent Urea-Based, Exosite-Binding Inhibitors Selective for Glutamate Carboxypeptidase II. Journal of medicinal chemistry 58, 4357-63; Weineisen, M., Simecek, J., Schottelius, M., Schwaiger, M., and Wester, H.-J. (2014) Synthesis and preclinical evaluation of DOTAGA-conjugated PSMA ligands for functional imaging and endoradiotherapy of prostate cancer. EJNMMI Res 4, 1-15.

[0250] Although the foregoing subject matter has been described in some detail by way of illustration and example for purposes of clarity of understanding, those skilled in the art will understand that changes and modifications may be practiced within the scope of the appended claims.

Claims

1. A complex compound of a compound of formula (I) with a radiometal selected from the group consisting of 212Pb, 225Ac, 213Bi, and 203Pb, and a pharmaceutically acceptable salt thereof, wherein the compound of formula (I) is 【Chemistry 1】 where: Z is CO 2 Q is; Q is H; m is an integer selected from the group consisting of 1, 2, 3, 4 and 5; R is -CH 2 -R 1 is; R 1 teeth, 【Chemistry 2】 is; wherein X is independently Br or I; L is C 1 -C 6 an alkylene linker; W is -(C=O)-NR 2 -is; R 2 is H; n is 1; Ch is a chelating agent having the following structure: 【Transformation 3】 and pharmaceutically acceptable salts thereof.

2. The compound of formula (I): 【Chemistry 4】 2. The complex compound of claim 1, selected from the group consisting of:

3. A pharmaceutical composition for treating tumors or cells that express one or more PSMAs, comprising the complex compound of claim 1 and a pharmaceutically acceptable salt thereof.

4. 4. The pharmaceutical composition of claim 3, wherein the one or more PSMA-expressing tumors or cells are selected from the group consisting of prostate tumors or cells, metastatic prostate tumors or cells, lung tumors or cells, kidney tumors or cells, glioblastoma, pancreatic tumors or cells, bladder tumors or cells, sarcoma, melanoma, breast tumors or cells, colon tumors or cells, germ cells, pheochromocytoma, esophageal tumors or cells, gastric tumors or cells, and combinations thereof.

5. The pharmaceutical composition of claim 3, wherein the one or more PSMA-expressing tumors or cells are prostate tumors or cells.

6. The pharmaceutical composition of claim 3, wherein the one or more PSMA-expressing tumors or cells are present in a subject.

7. The pharmaceutical composition of claim 6 , wherein the subject is a human.

8. The pharmaceutical composition of claim 3, wherein the pharmaceutical composition results in inhibition of the growth of the tumor.

9. The compound of formula (I) is 【Transformation 5】 where: M is 225 The complex compound according to claim 1, wherein Ac is a pharmaceutically acceptable salt thereof.

10. The compound of formula (I) is 【Transformation 6】 where: M is 225 The pharmaceutical composition of claim 3, wherein Ac is

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