Radiopharmaceutical composition and method

Abstract

To provide a method for preparing a radiopharmaceutical preparation.SOLUTION: Provided is a method for preparing a compound represented by the following formula (where Z1 is absent or is a radioactive cation), or a pharmaceutically acceptable salt thereof.SELECTED DRAWING: None

Description

Technical Field

[0001] There are provided radiopharmaceutical formulations and treatment methods including the treatment of cancer.

Background Art

[0002] Radiopharmaceuticals have been used in various therapeutic and diagnostic applications. In particular, radiolabeled molecules are useful for treating various malignancies including prostate cancer.

[0003] Prostate cancer is a common and lethal disease that can recur after initial treatment. Prostate cancer is the most common cancer in men in the United States and is the second leading cause of cancer-related death. Approximately one-third of patients who receive therapeutic treatment (radiation therapy or radical prostatectomy) experience an increase in the level of prostate-specific antigen (PSA), a biomarker for prostate cancer, which indicates biochemical recurrence (BCR).

[0004] PSMA-PET imaging has enabled the identification of oligometastatic hormone-sensitive prostate cancer (OmHSPC). OmHSPC is a pathological condition with few (usually five or fewer) metastatic lesions that can be detected by imaging. PSMA-PET is a new imaging modality that uses a radioactive tracer that binds to prostate-specific membrane antigen (PSMA), a protein overexpressed in most prostate cancer cells. PSMA-PET can detect lesions that are invisible on conventional imaging and has led to the stage migration from BCR to OmHSPC.

[0005] Androgen deprivation therapy (ADT) is the standard treatment for hormone-sensitive prostate cancer, but it has severe side effects and its effectiveness is limited. ADT is a systemic therapy that suppresses the androgen pathway, which is essential for the growth of prostate cancer cells. ADT is used for both BCR and OmHSPC, but it has many adverse effects on the patient's quality of life and long-term health (e.g., sexual dysfunction, metabolic complications, osteoporosis, and cardiovascular disease). Furthermore, most patients ultimately develop resistance to ADT and progress to castration-resistant prostate cancer (CRPC), which has a poor prognosis.

[0006] Androgen receptor pathway inhibitors (ARPI) are a new class of drugs that have shown effectiveness in CRPC but are not curative. ARPI are oral agents that target the androgen receptor or its ligand and have been approved for the treatment of CRPC in recent years. ARPI have improved the survival and quality of life of patients with CRPC, but they are not without side effects such as fatigue, rash, and hypertension. Furthermore, most patients develop resistance to ARPI within a few years and require additional treatment options.

[0007] The first newly approved treatment paradigm within the prostate cancer treatment space is the use of radioligand theranostic pairs to select and then treat patients who overexpress PSMA. This unique treatment approach utilizes the expression pattern of this target within prostate cancer to deliver damaging radiation directly to cancer cells throughout the body.

[0008] In addition, the manufacture of these pharmaceuticals presents various challenges, including those related to their yield and purity. For compounds with multiple chiral centers that require long synthetic routes, the yield not only decreases over multiple-step schemes but the purity is also often compromised. Furthermore, the configuration of the chiral centers is inverted in peptide synthesis, resulting in the formation of two or more product isomers. These unwanted isomers can lead to poor stability, a shortened shelf life, and a reduced therapeutic effect. SUMMARY OF THE INVENTION

[0009] The inventors herein provide a formulation containing, with high chemical purity, a compound of the following formula A-2:

[0010] [Chemical formula] (wherein Z is a radiopharmaceutical containing a chelating radioactive cation).

[0011] The inventors have also demonstrated herein the therapeutic use of the compounds disclosed herein in human subjects, including human patients suffering from prostate cancer.

[0012] In a preferred embodiment, an R enantiomer-enriched compound of the following formula A-2-R:

[0013] [Chemical formula] (wherein Z is a radiopharmaceutical containing a chelating radioactive cation), that is, a compound in which the R enantiomer of the carbon shown in formula A-2 is substantially enriched, is provided with high chemical purity. *

[0014] In a preferred embodiment, an S enantiomer-enriched compound of the following formula A-2-S:

[0015] [Chemical formula] (wherein Z is a radiopharmaceutical containing a chelating radioactive cation), that is, a compound in which the S enantiomer of the carbon shown in formula A-2 is substantially enriched, is provided with high chemical purity. *

[0016] In a particular embodiment, a compound of formula A-3 is provided with high chemical purity.

[0017] [Chemical formula] ​​

[0018] In a preferred embodiment, the compound of formula A-3-R is represented as follows:

[0019] [ka] R isomer-enriched compounds (i.e., compounds of formula A-3) * Compounds substantially enriched in the R isomer of carbon are provided in high chemical purity.

[0020] The compound of the above formula A-3-R is defined herein as 225 Also referred to as Ac PSMA-62. Preferably, the compound of formula A-3-R (i.e. 225 Ac PSMA-62) is used in a formulation or method of treatment in a high purity (e.g., greater than 90% chemical purity). The compound of formula A-3-R above, but not including Ac-225 or other radioisotopes chelated thereto, may also be referred to herein as PSMA-62 (shown as formula A-1 below), and is preferably of high purity (e.g., greater than 90% chemical purity).

[0021] In a preferred embodiment, the compound of formula A-3-S is as follows:

[0022] [ka] S-isomer enriched compounds (i.e., compounds of formula A-3) * Compounds substantially enriched in the S isomer of carbon are provided in high chemical purity.

[0023] The structures containing chelated metals are not intended to represent the exact binding mode of each metal. Rather, they are intended to illustrate where the metals are chelated. For example, 225 Ac is as follows:

[0024] [ka] It is considered to bind as (in the formula, the wavy line is the binding point of the chelating agent to the rest of the molecule).

[0025] In multiple aspects, as disclosed herein, methods and uses are provided for diagnosing and / or monitoring a disorder, including a cancer. In one aspect, these methods and uses may include administering a radiocontrast agent to a subject (e.g., a human patient). In multiple aspects, preferably, the radiocontrast agent is a radiocontrast agent that targets PSMA, for example, 68 Ga-PSMA-11 and 18 F-DCFPyl may be included. The radiocontrast agent may also include a radiolabeled compound as disclosed herein, such as a compound of formula A-2. In multiple aspects, the radiocontrast agent is 68 Ga, for example 68 a compound disclosed herein that is suitably complexed with Ga-PSMA-62. A radiocontrast agent, such as a compound disclosed herein, can be administered to a subject and the subject can be monitored. The protocol may include 68 allowing a radiocontrast agent, such as Ga-PSMA-62, to bind to the subject's tissue, and then monitoring the subject (especially outside the body), for example, by positron emission tomography (PET). Thereafter, based on the results of the diagnostic protocol, 225 a radiopharmaceutical agent as disclosed herein, such as Ac-PSMA-62, can be administered to treat a specified disease, including metastatic castration-resistant prostate cancer (mCRPC) and biochemical recurrence (BCR) prostate cancer. Also thereafter, based on the results of the diagnostic protocol, 225 a radiopharmaceutical agent as disclosed herein, such as Ac-PSMA-62, can be administered to treat a specified disease, including metastatic castration-resistant prostate cancer (mCRPC) and / or oligometastatic hormone-sensitive prostate cancer (OmHSPC).

[0026] In such diagnostic / monitoring methods and uses, preferred radiocontrast agents include 68 Ga-PSMA-11 and 18PSMA targeting agents such as F-DCFPyl, or in particular 68 compounds of the following formula B-11 complexed with a radioactive isotope of Ga:

[0027]

Chemical formula

[0028] In a preferred embodiment, an R enantiomer-enriched compound (i.e., a compound in which the R enantiomer of the carbon shown in formula B-11 is substantially enriched) can be used in a treatment method as disclosed herein, including a diagnostic or monitoring method. *

[0029]

Chemical formula

[0030] The compound of formula B-11-R above is also referred to herein as 68 Ga-PSMA-62 or B-11-R. Preferably, the compound of formula B-11-R (i.e., as Ga-PSMA-62) is used in high purity (e.g., greater than 90% chemical purity) in a formulation or treatment method (including a diagnostic or monitoring method). 68

[0031] In a preferred embodiment, the following formula B-11-S:

[0032]

Chemical formula

[0033] In one embodiment, the subject is identified for treatment by PSMA imaging, including PSMA-PET. ​​​

[0034] In a preferred embodiment, the composition of the present invention is administered to a subject identified as exhibiting a PSMA-positive disease or condition thereof, such as may be indicated by overexpression of PSMA in prostate cancer.

[0035] In certain embodiments, the subject has cancer that expresses prostate-specific membrane antigen (PSMA).

[0036] In one embodiment, the subject has or is suspected of having prostate cancer.

[0037] In one embodiment, the subject has or is suspected of having metastatic castration-resistant prostate cancer (mCRPC). In a further embodiment, the subject has metastatic castration-resistant prostate cancer (mCRPC) with prostate-specific membrane antigen (PSMA) avid lesions.

[0038] In one embodiment, the subject has or is suspected of having biochemically recurrent (BCR) prostate cancer.

[0039] In one embodiment, the subject has or is suspected of having oligometastatic hormone-sensitive prostate cancer (OmHSPC). In a further embodiment, the subject has oligometastatic hormone-sensitive prostate cancer (OmHSPC) with prostate-specific membrane antigen (PSMA) avid lesions.

[0040] In certain embodiments, the subject may be evaluated and / or identified as having high or elevated prostate-specific membrane antigen (PSMA) expression compared to a healthy subject, and a subject identified as having elevated PSMA activity is 225 administered a radiopharmaceutical compound disclosed herein that comprises Ac PSMA-62.

[0041] In certain embodiments, the method and use comprise imaging or other evaluation of the subject's cancer, including evaluation of PSMA expression. The evaluation is 225It can be appropriately performed before administering the radiopharmaceutical compounds disclosed herein containing Ac PSMA-62. The evaluation of cancer preferably includes imaging diagnosis, such as SPECT or PET imaging diagnosis, or immunohistochemistry / H4C or fluorescence in situ hybridization (FISH) imaging diagnosis. To facilitate cancer analysis, 18 Agent F, 68 Agent Ga, or 64 A contrast agent such as Agent Cu can be administered to the subject.

[0042] In one aspect, if a subject shows at least one PSMA-PET positive lesion of prostate cancer, it is identified for the treatment of mCRPC. In a certain aspect, the subject can also be determined to be PSMA positive according to the following criteria: i) all solid organ metastases (e.g., lungs, adrenal glands, etc.) with a short axis of 10 mm or more must be PSMA-PET positive; ii) all lymph nodes measured to be 25 mm or more in the short axis in anatomical imaging must be PSMA-PET positive; and iii) all bone metastases with a soft tissue component of 10 mm or more in the short axis are PSMA-PET positive.

[0043] In one aspect, if a subject shows 1 to 5 or more positive lesions outside the prostatic bed or residual gland, it is identified for the treatment of BCR prostate cancer.

[0044] In another aspect, a subject showing 1 to 5 or more positive lesions outside the prostatic bed or residual gland is identified for the treatment of OmHSPC.

[0045] In one aspect, if a subject shows 1) a PSMA expression standard uptake value of SUVmax of 15 or more at one site of the disease and / or 2) a PSMA expression standard uptake value of SUVmax greater than 10 at all measurable disease sites, the subject is identified for treatment.

[0046] In another aspect, if a subject shows at least one positive lesion SUVmax greater than 10, the subject is identified for treatment.

[0047] For patients receiving treatment for mCRPC, a preferred dosage may include administering Ac PSMA-62 once on the first day of each 6-week cycle over a total of 4 cycles. 225 Ac PSMA-62 may be administered once.

[0048] For patients receiving treatment for mCRPC, a preferred dosage may include administering Ac PSMA-62 once on the first day of each 6-week cycle. 225 Ac PSMA-62 may be administered once.

[0049] For patients receiving treatment for mCRPC, a preferred dosage may include administering Ac PSMA-62 one or more times (e.g., 2, 3, or 4 times) on the first day during a treatment cycle (e.g., a 2-week cycle, 3-week cycle, 4-week cycle, 5-week cycle, 6-week cycle, 7-week cycle, 8-week cycle, 9-week cycle, 10-week cycle, 11-week cycle, or 12-week cycle). 225 Ac PSMA-62 may be administered one or more times (e.g., 2, 3, or 4 times).

[0050] For patients receiving treatment for BCR prostate cancer, a preferred dosage may include administering Ac PSMA-62 once on the first day of each 8-week cycle over a total of 2 cycles. 225 Ac PSMA-62 may be administered once.

[0051] For patients receiving treatment for BCR prostate cancer, a preferred dosage may include administering Ac PSMA-62 once on the first day of each 8-week cycle. 225 Ac PSMA-62 may be administered once.

[0052] For patients receiving treatment for BCR prostate cancer, a preferred dosage may include administering Ac PSMA-62 one or more times (e.g., 2, 3, or 4 times) on the first day during a treatment cycle (e.g., a 2-week cycle, 3-week cycle, 4-week cycle, 5-week cycle, 6-week cycle, 7-week cycle, 8-week cycle, 9-week cycle, 10-week cycle, 11-week cycle, or 12-week cycle). 225 Ac PSMA-62 may be administered one or more times (e.g., 2, 3, or 4 times).

[0053] For patients undergoing treatment with OmHSPC, the preferred dosage may include administering Ac PSMA-62 once on the first day of each 8-week cycle over a total of 2 cycles. 225 It may include administering Ac PSMA-62 once.

[0054] For patients undergoing treatment with OmHSPC, the preferred dosage may include administering Ac PSMA-62 once on the first day of each 8-week cycle. 225 It may include administering Ac PSMA-62 once.

[0055] For patients undergoing treatment with OmHSPC, the preferred dosage may include administering Ac PSMA-62 one or more times (e.g., 2, 3, or 4 times) on the first day during a treatment cycle (e.g., a 2-week cycle, 3-week cycle, 4-week cycle, 5-week cycle, 6-week cycle, 7-week cycle, 8-week cycle, 9-week cycle, 10-week cycle, 11-week cycle, or 12-week cycle). 225 It may include administering Ac PSMA-62 one or more times (e.g., 2, 3, or 4 times).

[0056] In certain embodiments, the first treatment for the subject's cancer is 225 Treatment with a radiopharmaceutical disclosed herein that includes Ac PSMA-62.

[0057] In other embodiments, the subject 225 Has received treatment with one or more other cancer therapies prior to being administered a radiopharmaceutical disclosed herein that includes Ac PSMA-62. For example, the subject 225 May have received a surgical procedure prior to being administered a radiopharmaceutical disclosed herein that includes Ac PSMA-62. The subject 225 May have received other chemotherapy prior to being administered a radiopharmaceutical disclosed herein that includes Ac PSMA-62.

[0058] In one embodiment, the subject 225Before administration of the radiopharmaceuticals disclosed herein that include Ac PSMA-62, the subject may have been treated with one or more androgen receptor inhibitors. In one aspect, the subject is before treatment with one or more androgen receptor inhibitors, 225 Before administration of the radiopharmaceuticals disclosed herein that include Ac PSMA-62, the subject may be chemotherapy-naive.

[0059] In one aspect, the subject has mCRPC, 225 Before administration of the radiopharmaceuticals disclosed herein that include Ac PSMA-62, the subject has been treated with one or more androgen receptor inhibitors and / or androgen receptor pathway inhibitors and may have received (or been ineligible / declined) taxane chemotherapy; the subject may have received up to 3 previous systemic therapy regimens in the mCRPC setting.

[0060] In one aspect, the subject has prostate cancer with 1-5 PSMA-positive lesions and biochemical recurrence after radical surgery or radiation therapy and has not yet initiated lifelong hormonal therapy.

[0061] In one aspect, the subject has OmHSPC with 1-5 PSMA-positive lesions and biochemical recurrence after radical surgery or radiation therapy and has not yet initiated lifelong hormonal therapy.

[0062] This radiopharmaceutical formulation (pharmaceutical composition) is 225 A suitable aqueous composition comprising one or more radiopharmaceuticals contemplated herein that include Ac PSMA-62.

[0063] In some aspects, the pharmaceutical composition may be formulated for intravenous administration.

[0064] The pharmaceutical composition of the present invention may preferably include one or more additives such as one or more stabilizer compounds that can inhibit or prevent degradation of the prepared formulation.

[0065] In one aspect, the pharmaceutical composition can contain one or more ascorbate compounds, for example, in an amount of 10 to 50 mg of one or more ascorbate compounds per 1 mL of the aqueous pharmaceutical composition, or in an amount of 20 to 40 mg, or 24 to 26 mg of one or more ascorbate compounds per 1 mL of the aqueous pharmaceutical composition.

[0066] In one aspect, the pharmaceutical composition of the present invention can further contain a chelating agent that is added after the formation of the radiopharmaceutical compound and is suitable for removing radionuclides that do not form a complex. Suitable chelating agents can include, for example, one or more aminopolycarboxylic acids, such as diethylenetriaminepentaacetic acid (DTPA) or salts thereof, preferably in an amount that provides a concentration of 0.01 to 0.50 mg / mL in the aqueous formulation.

[0067] A method for treating a subject suffering from cancer, comprising: a) administering to the subject a therapeutically effective amount of one or more androgen receptor inhibitors; b) identifying the subject as showing cancer progression after administering one or more androgen receptor inhibitors; and then c) administering to the identified subject an effective amount of 225 the radiopharmaceutical disclosed herein, comprising Ac PSMA-62.

[0068] A method for treating a subject suffering from cancer, comprising: a) administering to the subject a therapeutically effective amount of one or more androgen receptor inhibitors, taxane chemotherapy, and up to three prior systemic therapy regimens in the context of the cancer situation; b) identifying the subject as showing cancer progression after administering one or more androgen receptor inhibitors; and then c) administering to the identified subject an effective amount of 225 the radiopharmaceutical disclosed herein, comprising Ac PSMA-62.

[0069] In a further aspect, a method for treating a subject suffering from prostate cancer, comprising: a) identifying a subject showing progression of prostate cancer during or after treatment with other cancer therapies, such as administration of one or more androgen receptor inhibitors; and b) administering to the identified subject an effective amount of 225 a radiopharmaceutical disclosed herein, comprising Ac PSMA-62. A method is provided that includes these steps.

[0070] In a further aspect, a method for treating a subject suffering from prostate cancer, comprising: a) identifying a subject showing progression of prostate cancer during or after treatment with other cancer therapies, such as administration of one or more androgen receptor inhibitors, taxane chemotherapy, and up to three previous systemic therapy regimens in the cancer situation; and b) administering to the identified subject an effective amount of 225 a radiopharmaceutical disclosed herein, comprising Ac PSMA-62. A method is provided that includes these steps.

[0071] In one aspect of the above method, the subject shows cancer progression during treatment with one or more of abiraterone, enzalutamide, apalutamide, darolutamide, cimetidine, orteronel, galeterone, seviteronel, toppicalutamide, bicalutamide, fluamide, and / or nilutamide. In a further aspect, the subject shows cancer progression during treatment with abiraterone, enzalutamide, apalutamide, and / or darolutamide.

[0072] In a further aspect, the subject is identified for treatment if the subject has elevated prostate-specific antigen (PSA) levels, such as PSA levels of 3 ng / mL, 4 ng / mL, 5 ng / mL, or 6 ng / mL or higher. In certain embodiments, administration of one or more androgen receptor inhibitors is 225 completed prior to administration of a radiopharmaceutical disclosed herein, comprising Ac PSMA-62.

[0073] In certain embodiments, the treatment method and therapeutic use may include subjects having one or more of the following specified features (inclusion criteria): 1. Male patients of a specific age, such as at least 18 or 21 years old; 2. Female patients of a specific age, such as at least 18 or 21 years old. 3. Histological, pathological, and / or cytological confirmation of prostate adenocarcinoma; 4. Appropriate organ function (bone marrow reserve, liver function, kidney function); 5. Human immunodeficiency virus patients who are healthy and at low risk of acquired immunodeficiency syndrome-related outcomes; 6. Patients who are willing and able to comply with all study requirements and treatments ( 225 including Ac PSMA-62) and the timing and nature of the required evaluations; 7. ECOG performance status of 0-2; 8. ECOG performance status of 0-1; 9. A life expectancy of at least 6 months as determined by the treating investigator; 10. The patient and their female partner of childbearing potential must agree to use two acceptable forms of contraception, one of which must be a barrier method during the study and for 21 weeks after the last dose of study drug. 11. The patient has read, understood, and signed the written informed consent form; 12. Criteria specific to mCRPC patients: a. They have previously received treatment for their underlying disease and have exhausted all satisfactory or available approved treatment options; b. Have progressive mCRPC at the time of consent based on at least one of the following criteria: · Serum / plasma PSA progression defined as an increase in PSA that exceeds the nadir by more than 25% and more than 2 ng / mL, confirmed by a PSA test at least one week later; · PSA progression defined as an increase in PSA values at a minimum one-week interval with a final result of at least 1.0 ng / mL · Soft tissue progression defined as an increase of 20% or more in the sum of the diameters (sum of the diameter, SOD) of all target lesions based on the smallest SOD since the start of treatment (short axis for nodular lesions and long axis for non-nodular lesions), or the appearance of one or more new lesions; and / or · Progression of bone disease defined as the appearance of one or more new lesions on bone scan; c. PSMA-PET positivity within 90 days of enrollment; note that either [Ga-68] or [F-18] PSMA targeting agent, which has been approved for commercial use, can be used according to the diagnostic standard treatment; d. Castration-circulating testosterone level (less than 1.74 nmol / L or less than 50 ng / dL). 13. Criteria specific to BCR patients: a. The patient's primary tumor must not have been previously treated with surgery and / or radical radiation. Previous salvage treatment (radiation or surgery) to the prostatic bed or pelvis is acceptable; b. Biochemical recurrence after primary treatment. Patients who have not previously received ADT, or patients who have received prior ADT and have testosterone recovered within the normal range defined as 5.2 nmol / L or higher (150 ng / dL or higher) must meet one or more of the following thresholds for BCR: · Patients who have previously undergone radical prostatectomy, with or without radical radiation therapy, must have a prostate-specific antigen (PSA) of 0.2 ng / mL or higher, or · Patients who have received only radical radiation for the treatment of the primary tumor must have a PSA that exceeds the nadir by 2 ng / mL or more, c. Positive PSMA-PET within 90 days of enrollment. Either [Ga-68] or [F-18] PSMA targeting agent approved by each healthcare institution can be used according to the institution's standard treatment (diagnostic protocol); d. No indication of urgent or emergency radiation; e. The patient has not received any form of prostate cancer-directed therapy since undergoing the screening PSMA scan. 14. Criteria specific to OmHSPC patients: a. PSA recurrence after radical prostatectomy (RP) or radical radiotherapy (RT), with or without (neo)adjuvant ADT, with or without adjuvant / salvage local therapy (radiation or surgery) · For patients who have previously undergone RP + / - RT, PSA is 0.2 ng / mL or higher, or · For patients who have previously undergone RT only, PSA exceeds the nadir by 0.2 ng / mL or more b. 1 - 5 PSMA_PET positive lesions identified outside the prostate bed or residual gland.

[0074] In some embodiments, one or more of the above incorporation criteria may be appropriately presented on the label, or instructions for use, or otherwise, 225 It may be associated with a kit, pharmaceutical composition, or other delivery package, system, or presentation of a radiopharmaceutical agent discussed herein that includes Ac PSMA - 62.

[0075] In certain embodiments, the present treatment methods and therapeutic uses may exclude subjects having one or more of the following specified characteristics (exclusion criteria): Exclusion criteria: 1. Patients with neuroendocrine or small cell cancer of the prostate; 2. Have undergone major surgery within 30 days prior to the start of the trial; 3. The patient is receiving any other investigational therapeutic agent within 4 weeks or 5 half - lives (whichever is shorter) from the start of the trial; 4. Evidence of ongoing and untreated urinary obstruction; 5. History of grade 4 myelosuppression lasting more than 7 days, or grade 3 myelosuppression requiring more than 6 weeks to recover; 6. Patients who are receiving a drug known to cause xerostomia or xerophthalmia (e.g., darifenacin) are excluded if they have not been on a stable dose for at least 4 weeks prior to screening; 7. Any reason for existing grade 1 oral dryness (xerostomia) or symptomatic grade 1 xerophthalmia (dry eye); 8. Contraindications to the use of the planned [Ac-225]-PSMA-62 therapy, including but not limited to hypersensitivity to the [Ac-225]-PSMA-62 excipients; 9. Having a known history of another malignancy within the past 5 years. Exceptions: Malignancies that have been cured by treatment and have not recurred within 2 years prior to the study; completely resected basal cell carcinoma and squamous cell skin carcinoma; any malignancy considered to be asymptomatic and that has never required treatment; and any type of cancer completely resected in situ; 10. The patient has any concurrent severe and / or uncontrolled medical condition that may increase the patient's risk of toxicity during the study or may confound the distinction between the disease and study-related toxicity; 11. Severe psychological, family, social, or geographical conditions that may prevent compliance with the study protocol and follow-up schedule; 12. Symptomatic spinal cord compression, or clinical or radiological findings indicating impending spinal cord compression; 13. Inability to lie flat or tolerate PET / CT or MRI during the procedure; 14. History of torsade de pointes or congenital long QT syndrome; 15. Concurrent severe (as determined by the study responsible physician) medical condition; 16. Criteria specific to mCRPC patients: a. The patient has received any therapeutic systemic radionuclide (e.g., radium-223, rhenium-186, strontium-89), or therapeutic radioligand (e.g., Lu-177-PSMA) within 5 half-lives of the start of the study, b. Patients currently receiving systemic anti-cancer therapy, except for ADT with or without ARPI. However, patients may be included in the trial if they discontinue all prohibited anti-cancer therapies before receiving the first dose of the investigational drug. c. Since the PSMA-PET scan has been used for eligibility, the patient has started treatment with an alternative ARPI or switched to an alternative ARPI. Patients already receiving ARPI may continue treatment with the same ARPI during the trial. d. Patients with a history of central nervous system (CNS) metastases must have received treatment (surgery, radiotherapy, gamma knife), must be neurologically stable, and must not be receiving corticosteroids for the purpose of maintaining nerve integrity. Patients with epidural disease, spinal canal disease, and previous spinal cord involvement are eligible if those areas have been treated and are stable. For patients with substantial CNS metastases (or a history of CNS metastases), baseline and subsequent radiological imaging must include an evaluation of the brain. 17. Criteria specific to BCR patients: a. Any previous cytotoxic chemotherapy; b. Prior treatment with a therapeutic systemic radionuclide (e.g., radium-223, rhenium-186, strontium-89), or a therapeutic radioligand (e.g., Lu-177-PSMA); c. Previous immunotherapy or adoptive T cell therapy (e.g., CAR-T therapy, TCR therapy, etc.); d. A previous poly ADP-ribose polymerase (PARP) inhibitor for prostate cancer; e. The patient is receiving any systemic anti-cancer therapy for prostate cancer, except for a limited course of ADT for the management of localized disease; f. The presence of any liver metastases; g. Use of opioids for cancer-related pain within 30 days prior to consent; h. Known presence of central nervous system metastases. 18. Criteria specific to OmHSPC patients: a. The patient is receiving some systemic anti-cancer therapy for prostate cancer, except for (neo)adjuvant ADT for the management of localized disease. b. The presence of any liver metastases c. The known presence of central nervous system metastases. d. The patient had PSA progression in relation to castrate levels of testosterone.

[0076] In some embodiments, one or more of the above exclusion criteria may be appropriately presented on the label or instructions for use, or otherwise, 225 Ac PSMA-62 may be associated with a kit, pharmaceutical composition or other delivery package, system or presentation of a radiopharmaceutical agent discussed herein.

[0077] In some embodiments, the compounds of formula A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S are provided at a purity greater than 90%, or greater than 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5%. Such purity levels can be defined as chemical purity and can be determined by chromatography, particularly UHPLC. As will be understood, the chemical impurities referred to herein may or may not contain radioisotopes such as actinium-225 or ruthenium-177 or their degradation products. In certain embodiments, the chemical impurities may be referred to herein as one or more compounds or compositions that do not contain radioisotopes such as actinium-225 or ruthenium-177 or their degradation products.

[0078] In certain embodiments, a compound of any of formulae A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S is provided in the substantial absence of structural (including constitutional and / or stereoisomers, e.g., diastereoisomers) impurities. The term "structural impurities" refers to impurities that have the same or substantially or essentially the same (e.g., at least 80, 85, 90, 95, 96, 97, 98, or 99 percent) molecular weight and composition (structure) as a particular compound of formulae A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S, and the impurities may have differences in the connectivity or stereochemical arrangement between one or more atoms as compared to the particular compound.

[0079] In one embodiment, "structural impurities" are referred to herein with respect to compounds having a retention time (peak) within, at most, or less than 4, 3, 2.8, 2.6, 2.4, 2.2, 2.0, 1.8, 1.6, 1.4, 1.2, 1.0, 0.8, 0.6, 0.4, 0.2, or 0.1 minutes of the peak of the desired compound of formula A, by high performance liquid chromatography (HPLC including UHPLC), specifically an Agilent 1290 / 6460 Triple Quad LC / MS system equipped with a UV detector and a Waters XBridge BEH Phenyl Column, 4.6×150 mm, 3.5 μm. The gradient elution system utilizes mobile phase A (0.05% TFA in HPLC grade water, pH 2.02) and mobile phase B (ACN); the gradient starts at 85% A and 15% B and is increased to 76% A and 24% B over 20 minutes; then it is increased to 64% A and 36% B over 4 minutes, and can be run at a flow rate of 1.0 mL / min, followed by returning the gradient parameters to the initial starting conditions.

[0080] In certain embodiments, the "structural impurities" referred to herein with respect to compounds of any of Formulae A-1, A-2, or A-3 are characterized as having retention times (peaks) in the region of 13.6 to 14.2 minutes, or 13.8 to 14.0 minutes, particularly 13.9 minutes, by an Agilent 1290 / 6460 Triple Quad LC / MS system equipped with a UV detector (monitored at 210 nm) using high performance liquid chromatography (HPLC), particularly a Waters XBridge BEH Phenyl Column, 4.6×150 mm, 3.5 um (exemplified by the chromatograms of Example 4 and Figure 3 below). The gradient elution system utilizes mobile phase A (0.05% TFA in HPLC grade water, pH 2.02) and mobile phase B (ACN); the gradient starts at 85% A and 15% B and is increased to 76% A and 24% B over 20 minutes; then it is increased to 64% A and 36% B over 4 minutes, and can be run at a flow rate of 1.0 mL / min, followed by returning the gradient parameters to the initial starting conditions.

[0081] In certain embodiments, one or more structural impurities are present in an amount of 10%, 9%, 8%, 7%, 6%, 5%, 4.0%, 3.8%, 3.6%, 3.4%, 3.2%, 3.0%, 2.8%, 2.6%, 2.4%, 2.2%, 2.0%, 1.0% or 0.5% or less, as determined by HPLC including, for example, UHPLC and / or mass spectrometry, with specific compounds of Formulae A-2, A-2-R, A-2-S, A-3, A-3-R and / or A-3-S. Such % values of one or more structural impurities as referred to herein in the case of HPLC including UHPLC analysis can refer to % of the total area of the chromatogram (which may also be referred to as radiochemical purity). Such % values of one or more structural impurities as referred to herein can also be weight % based on the total weight of the specific compound of a specific compound of Formulae A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S.

[0082] In some embodiments, the structural impurities that may be associated with (and present in an amount less than 10%) any of the compounds of formula A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S may include one, two, three, or more of the compounds of the following formulas B-1, B-1-R, B-1-S, B-2, B-2-R, B-2-S, B-3, B-3-R, and B-3-S. The structural impurities that may be associated with (and present in an amount less than 10%) any of the compounds of formula B-11, B-11-R, and / or B-11-S may include one, two, three, or more of any of the compounds of the following formulas B-1, B-1-R, B-1-S, B-2, B-2-R, B-2-S, B-3, B-3-R, and B-3-S. In some embodiments, the structural impurities that may be associated with (and less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4.0%, less than 3.8%, less than 3.6%, less than 3.4%, less than 3.2%, less than 3.0%, less than 2.8%, 2.6%, 2.4%, 2.2%, 2.0%, 1 or 0.5% or less, for example, present in an amount determined by HPLC including the conditions defined above) any of the compounds of formula A-2, formula A-2-R or formula A-2-S may include one, two, three, or more of the compounds of the following formulas B-1, B-2 or B-3.

[0083] [Chemical Formula]

[0084] [Chemical Formula]

[0085] [Chemical Formula]

[0086] In some embodiments, in any of the compounds of formula A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S, the amounts of such impurities B-1, B-2 and B-3 are less than 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1%, as can be determined by chromatography, particularly UHPLC, including under the conditions specified above. Such purity levels (i.e., chemical purity greater than 90% and / or substantial absence of impurities such as structural impurities) of any of the compounds of formula A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S can be provided by the reaction products of the syntheses disclosed herein (see Examples 1 and 2), and formulations of such reaction products without further processing steps such as chromatography (particularly purification). Thus, importantly, in certain embodiments, the compound reaction products can be directly formulated and packaged (e.g., stored in sealed vials or IV bags) after synthesis at such high purity levels without the need for purification (e.g., chromatography) or other processing steps to remove impurities.

[0087] In some embodiments, after release of the compounds disclosed herein from the resins according to these syntheses disclosed herein, particularly Examples 1 and 2 below, without additional purification steps, the structural impurity content of the compounds is determined to be less than 10%, less than 9, 8, 7, 6 or 5%, less than 4, 3 or 2%, or less than 1% based on the total amount or weight of the compound sample, for example, by UHPLC. As discussed herein, such percent presence or absence of structural impurity content can be appropriately determined by HPLC including UHPLC and / or mass spectrometry.

[0088] In a further aspect, radioisotopes, such as 177 Lu, 225 Ac, 211 At, 64 Cu, 67 Cu, 161 Tb, 67 Ga, 68 Ga, 203 Pb, 223Ra and / or 212 Compounds of formulae A-2, A-2-R, and A-2-S that are complexed or chelated with Pb are provided. In certain embodiments, the compounds of formulae A-2, A-2-R, and A-2-S are 225 Ac,[[]] 68 Ga, or 177 complexed with Lu. In one preferred embodiment, the compounds of formulae A-2, A-2-R, and A-2-S are 225 complexed with Ac.

[0089] Also provided are methods of treatment comprising administering to a subject suffering from a cell proliferative disease or disorder, particularly cancer, an effective amount of a compound disclosed herein comprising a compound of formulae A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S.

[0090] In particular, the compounds and compositions of the invention are useful for treating subjects suffering from metastatic castration-resistant prostate cancer (e.g., that can arise due to progression of the disease despite prior surgical or chemical castration), and / or biochemical recurrence (BCR) prostate cancer, and / or oligometastatic hormone-sensitive prostate cancer (OmHSPC).

[0091] Also provided is the use of the compounds and compositions for treating patients (such as humans) suffering from cancer.

[0092] In some embodiments, a subject being treated with a radiopharmaceutical disclosed herein can be evaluated by one or more of the following outcome measures: 1. 225 Adverse events (TEAE) and dose-limiting toxicities (DLT) occurring during treatment with Ac PSMA-62 [e.g., time frame: from the first administration of the investigational drug until the end of treatment (about 16 - 24 weeks)]; 2. 225 Changes in laboratory values, vital signs, and physical examinations after treatment with Ac PSMA-62 [e.g., time frame: from the first administration of the investigational drug until about 5 years]; 3. Time to initiation of any lifetime ADT or hormonal therapy [e.g., time frame: from first administration of the drug up to about 5 years]; 4. Objective response rate (ORR) for mCRPC only: Preliminary efficacy assessment based on Response Evaluation Criteria in Solid Tumors (RECIST 1.1) (soft tissue) and Prostate Cancer Working Group 3 (PCWG3) criteria (bone) [e.g., time frame: from first administration of the investigational drug until disease progression (up to about 3 years maximum)]; 5. Absorbed dose estimates (Gy) in normal organs: 225 Evaluation of the biodistribution and dosimetry of Ac PSMA - 62 in normal organs [e.g., time frame: from first administration of the investigational drug until the end of treatment (e.g., about 16 - 24 weeks)]; 6. PSA decrease from baseline: 225 To determine the effect of Ac PSMA - 62 on prostate - specific antigen (PSA) kinetics [e.g., time frame: from first administration of the investigational drug until the efficacy follow - up period (up to about 3 years maximum)]; 7. Change from baseline in PRO - CTCAE questionnaire score: To determine the effect of [Ac - 225] - PSMA - 62 on patient - reported outcomes (PRO) related to symptomatic adverse events.

[0093] In a further aspect, a kit containing the compounds or compositions disclosed herein is provided. In such a kit, in one aspect, the compound, e.g., the A - 2, A - 2 - R, A - 2 - S, A - 3, A - 3 - R, and / or A - 3 - S compounds that do not contain a radioisotope chelated thereto, is in a vial or other container in lyophilized form, or 177 Lu, 225 Ac, 211 At, 64 Cu, 67 Cu, 161 Tb, 67 Ga, 68 Ga, 203 Pb, 223 Ra, and / or 212It can be provided in other forms separated from radioactive isotopes such as Pb. In this embodiment, the radioactive isotope is reacted with the compounds of A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S that are not chelated with the radioactive isotope in a medical facility to provide a complexed radiopharmaceutical agent, which can then be administered to a patient. Such kits containing cold kits may include components such as one or more buffers such as acetic acid compounds and / or one or more radiation protectants or stabilizers.

[0094] In yet another embodiment, a packaged preparation or product of a radiopharmaceutical compound or composition is provided. The packaged preparation may include compounds of A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S, and optionally, 2) instructions for using the radiopharmaceutical compound or composition for treating cancers such as prostate cancer. Preferably, the packaged preparation contains a therapeutically effective amount of the radiopharmaceutical compound or composition. The instructions may appropriately be in written form including a package label. The radiopharmaceutical compound or composition may be appropriately contained in a lead container or other container within a further package containing product identification, instructions for use, or other information.

[0095] In a further embodiment, a treatment kit is provided. In a preferred embodiment, the kit comprises a) a pharmaceutical preparation of a radiopharmaceutical compound disclosed herein, including an effective amount of 225 Ac PSMA-62, sufficient to provide an effective treatment, suitable for intravenous administration to a patient, for example; and b) a package insert including instructions for the use of the agent in the treatment of a subject presenting with prostate cancer, including cancer, particularly metastatic castration-resistant prostate cancer (mCRPC) and / or biochemical recurrence (BCR) prostate cancer and / or OmHSPC.

[0096] In a more preferred embodiment, the kit comprises: a) a pharmaceutical preparation of a radiopharmaceutical compound disclosed herein, comprising an amount of 225Ac PSMA-62 sufficient to provide an effective treatment, for example, suitable for intravenous administration to a patient; and; and b) a package insert comprising instructions for the use of the agent in the treatment of a subject exhibiting prostate cancer, including cancer, particularly prostate-specific membrane antigen (PSMA)-positive metastatic castration-resistant prostate cancer (mCRPC) and / or biochemical recurrence (BCR) prostate cancer and / or oligometastatic hormone-sensitive prostate cancer (OmHSPC).

[0097] In another aspect, a method for making a compound of formula A-1:

[0098]

Chemical formula

[0099]

Chemical formula

[0100]

Chemical formula

[0101]

Chemical formula

[0102]

Chemical formula

[0103] In another aspect, the formula:

[0104] [Chemical formula] and its pharmaceutically acceptable salts are disclosed herein.

[0105] In yet another aspect, a method for preparing a compound of the formula:

[0106] [Chemical formula] (wherein Z1 is absent or is a radioactive cation) or a pharmaceutically acceptable salt thereof is disclosed herein.

[0107] In yet another aspect, a method for preparing a compound of the formula:

[0108] [Chemical formula] (wherein R 25 is the chelating agent described in FIG. 1 or FIG. 3) or a pharmaceutically acceptable salt thereof is disclosed herein, and the method comprises coupling a compound of the formula:

[0109] [Chemical formula] with the free acid form of the chelating agent group of FIG. 1 or FIG. 3, and the method comprises using a coupling agent in the presence of a solvent.

[0110] In another aspect, for formula M:

[0111] [Chemical formula] (wherein R 25 is the chelating agent described in FIG. 1 or FIG. 3) a method for preparing a compound or a pharmaceutically acceptable salt thereof is disclosed herein, and the method is as follows:

[0112] [Chemical formula] The compound of is coupled in the presence of a coupling agent and a coupling solvent (these terms are as defined herein) with formula Z:

[0113] [Chemical formula] This includes hydrolyzing the compound of formula M using the methods described herein to prepare a compound of the formula:.

[0114] [Chemical formula]

[0115] Other aspects and embodiments of the present invention are disclosed below. [Brief Description of the Drawings]

[0116] This patent document or this application document includes at least one drawing created in color. Copies of this patent or this patent application publication including color drawings will be provided by the Patent Office upon request and payment of the required fees.

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Mode for Carrying Out the Invention

[0117] The inventors herein provide, for the first time, formulations of compounds of formula A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S that do not contain radioisotopes chelated with them, have a high chemical purity level (e.g., a formulation chemical purity exceeding 90%), and have substantially no structural impurities.

[0118] The following text may refer to or exemplify specific embodiments of a compound or a method for the production of a compound, but is not intended to limit the scope of the compound or method to such specific references or examples. Various modifications can be made by those skilled in the art considering practical and economic considerations such as specific substituents of the compound and reagents and solvents for the synthesis of the compound.

[0119] As used herein, the articles "a" and "an" refer to "one or more" or "at least one" unless otherwise indicated. That is, a reference to any element or component of an embodiment by the indefinite article "a" or "an" does not exclude the possibility that two or more elements or components are present.

[0120] As used herein, the term "about" refers to plus or minus 10% of the referenced numerical indication. In some embodiments, "about" refers to plus or minus 5% of the referenced numerical indication.

[0121] The terms "chelating" or "chelating agent" are used interchangeably herein and refer to a molecule, often an organic molecule, often a Lewis base, having two or more non-shared electron pairs available for donation to a metal ion. The metal ion is typically coordinated to the chelating agent by two or more electron pairs. The terms "bidentate chelating agent", "tridentate chelating agent", and "tetradentate chelating agent" refer to chelating agents having two, three, and four electron pairs, respectively, readily available for simultaneous donation to a metal ion coordinated by the chelating agent. Usually, the electron pairs of the chelating agent form a coordination bond with a single metal ion. However, in certain examples, the chelating agent may form a coordination bond with two or more metal ions, and various bonding modes are possible.

[0122] As used herein, the term "purity" refers to the amount of a particular compound relative to the total amount of all isomers having the same formula as the particular compound. Purity can be measured by HPLC, UHPLC, or NMR.

[0123] As used herein, the term "chiral purity" refers to the purity of a particular compound relative to the total amount of its enantiomer or all diastereomers of the particular compound added together.

[0124] As used herein, the term "optically enriched" indicates the presence of one or more non-racemic stereocenters in a molecule, and the configuration of at least one stereocenter is such that one stereoisomeric configuration (R or S) is dominant. For example, one stereocenter in a molecule, typically a carbon atom, may have its bonded atoms spatially arranged in the (R) configuration in more than 50% by weight (based on the total weight of the compound). Alternatively, more than 50% by weight (based on the total weight of the compound) may be spatially arranged in the (S) configuration. More preferably, the molecule or its stereocenter is substantially optically enriched, and even more preferably, it is substantially enantiomerically pure.

[0125] As used herein, the term "substantially optically enriched", when referring to enantiomers or enantiomeric centers, means that at least about 60% by weight, preferably about 70% by weight, more preferably about 80% by weight, even more preferably about 90% by weight (based on the total weight of the compound) of one enantiomer or one enantiomeric center configuration is predominant in the mixture, and at least about 95% by weight (based on the total weight of the compound) of one enantiomer or one enantiomeric center configuration is even more preferred. In some preferred embodiments, the compound is "substantially enantiomerically pure", i.e., at least about 97.5% by weight, more preferably about 99% by weight, even more preferably about 99.5% by weight (based on the total weight of the compound) of one enantiomer configuration is predominant.

[0126] As used herein, the term "substantially pure" means being homogeneous enough to appear free of readily detectable impurities as determined by standard analytical methods such as thin layer chromatography (TLC), gel electrophoresis, high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR), and mass spectrometry (MS), or being pure enough such that further purification does not detectably alter the physical and chemical properties, or the biological and pharmacological properties, such as the enzymatic and biological activities, of the substance. In certain embodiments, "substantially pure" refers to a collection of molecules where at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 97%, about 98%, about 98.5%, about 99%, about 99.5%, or about 99.9% or more of the molecules, as determined by standard analytical methods, is a single compound containing a racemic mixture or one of its single enantiomers.

[0127] In one embodiment, the compounds disclosed herein, such as the compounds of Formula A-1 or Formula Y, etc., are optically enriched. In another embodiment, the compounds disclosed herein (such as the compounds of Formula A-1 or Formula Y, etc.) are substantially optically enriched. In another embodiment, the compounds disclosed herein (such as the compounds of Formula A-1 or Formula Y) are substantially pure.

[0128] The compounds disclosed herein are preferably single epimers at each chiral center, although other epimers are often present. For example, Formula Y uses an R-DOTAGA chelating group, but there is a high likelihood that at least a small amount of the S-DOTAGA chelating group is present.

[0129] The term "pharmaceutically acceptable carrier" refers to a chemical compound that facilitates the delivery or uptake of a compound or therapeutic agent into cells or tissues.

[0130] The term "pharmaceutical composition" refers to a mixture of a compound disclosed herein and other chemical components such as diluents or additional carriers. Pharmaceutical compositions facilitate the administration of the compound to an organism.

[0131] The term "coupling agent" refers to a compound that facilitates the formation of amide bonds. The coupling agent may or may not contain phosphorus in the molecule or counterion. Examples of phosphorus-containing coupling agents include (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), benzotrizylyloxy-tris[pyrrolidino]-phosphonium hexafluorophosphate (PyBOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP), hexafluorophosphate azabenzotriazole tetramethyluroniumnium (HATU), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT), but are not limited thereto. Examples of coupling agents that do not contain phosphorus include 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 1-hydroxy-7-azabenzotriazole (HOAT), or ethyl cyano(hydroxyimino)acetate (oxyma), diisopropylcarbodiimide (DIC), and dicyclohexylcarbodiimide (DCI). In some embodiments, a mixture of two or more coupling agents may be used. In other embodiments, other compounds such as a base, preferably a nitrogen-containing base, are present during the coupling reaction. Examples of nitrogen-containing bases include N,N-diisopropylethylamine (Hünig's base) and triethylamine. A preferred coupling agent is PyAOP in combination with N,N-diisopropylethylamine.

[0132] A variety of different solvents can be used in the coupling reactions described herein. Polar aprotic solvents are generally used. Examples of suitable solvents include chloroform, dichloromethane, and dimethylformamide (DMF). DMF is a preferred solvent for the couplings disclosed herein. The solvent used in the coupling reaction is referred to as the "coupling solvent".

[0133] In the hydrolysis reaction described in this specification, various acids can be used. The acid must be able to cleave the group to be cleaved. Examples of suitable acids include strong acids that can be used in the hydrolysis reaction described in this specification. Suitable acids include trifluoroacetic acid, hydrochloric acid, or para-toluenesulfonic acid. Other strong acids known in the art are also suitable.

[0134] The solvent used in the hydrolysis reaction is referred to as a "hydrolysis solvent". The hydrolysis solvent is independently selected each time it appears. A common solvent is water. Optionally, other co-solvents may be used. The solvent may contain additional additives such as triisopropylsilane or dithiothreitol. In some preferred embodiments, the solvent is water containing both triisopropylsilane (TIS) and dithiothreitol (DTT).

[0135] The term "amine protecting group" refers to any group known in the art of solid-phase peptide synthesis for protecting an amine from unwanted reactions. The amine protecting group should be selectively removable in the presence of other protecting groups. The amine protecting group is independently selected each time it appears. Examples of amine protecting groups include 9-fluorenylmethoxycarbonyl (Fmoc), benzyl, tert-butoxycarbonyl (Boc), benzyl chloroformate, trityl, 4-chlorotrityl, 4-methyltrityl (Mtt), N-[1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl] (Dde)

[0136] [Chemical formula] and 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl (IVdde), where the wavy line is the point of attachment. Fmoc and 4-methyltrityl are two preferred amine protecting groups.

[0137] The exact conditions required to remove an amine protecting group depend on the protecting group. For example, to remove an Fmoc group, a base such as cyclohexylamine, ethanolamine, piperidine, piperazine, triethylamine, or N,N - diisopropylethylamine is used in a polar aprotic solvent. The base used to remove the protecting group is referred to as an "amine deprotection base". The amine deprotection base is independently selected each time it appears. Examples of suitable solvents include dimethylformamide (DMF), dichloromethane, toluene, or N - methyl - 2 - pyrrolidone (NMP). Preferred solvents are DMF or CH2Cl2. More preferred is DMF. Such solvents are referred to as "amine deprotection solvents" or "amine deprotection solvents". The amine deprotection solvent is independently selected each time it appears. A preferred method for removing an Fmoc group is to treat it with piperidine in DMF. A preferred reagent for removing a 4 - methyltrityl group is hexafluoro - 2 - isopropanol.

[0138] Examples of carboxyl protecting groups include allyl, benzyl, tBu, and benzhydryl. Preferred carboxyl protecting groups are tBu or allyl. The carboxyl protecting group is independently selected each time it appears.

[0139] The exact conditions necessary to remove a carboxyl protecting group depend on the protecting group. For allyl, a preferred method for removing a carboxyl protecting group is to use a Pd-containing compound. A preferred Pd-containing compound is Pd(PPh3)4. The compound used to remove a carboxyl protecting group is referred to as a "carboxyl deprotecting agent". The carboxyl deprotecting agent is independently selected each time it appears. When removing an allyl protecting group, a nucleophile is generally present. Examples of suitable nucleophiles include 1,3-dimethylbarbituric acid and triphenylphosphine. The nucleophile is independently selected each time it appears. A preferred nucleophile is 1,3-dimethylbarbituric acid. A preferred method for removing an allyl, carboxyl protecting group is to use Pd(PPh3)4 in dichloromethane and dimethylformamide in the presence of 1,3-dimethylbarbituric acid. The tBu protecting group can be removed using methods known in the art such as hydrolysis. In one embodiment, the tBu group is removed using a strong acid in a solvent containing water.

[0140] The solvent used for carboxyl deprotection contains at least one of water, dichloromethane, dimethylformamide, tetrahydrofuran, or ethanol. Preferred solvents include at least one of water, dichloromethane, and dimethylformamide. These solvents are referred to as "carboxyl deprotection solvents". The carboxyl deprotection solvents are independently selected each time they appear.

[0141] Examples of oxygen protecting groups include methyl, t-butyl, and t-butyldimethylsilyl groups. A preferred oxygen protecting group is t-butyl, which is also specified herein as "tBu". The oxygen protecting groups are independently selected each time they appear.

[0142] The exact conditions necessary to remove the oxygen protecting group depend on the protecting group. A preferred method for removing the t-butyl oxygen protecting group is to use a strong acid. A preferred strong acid is TFA. The compound used to remove the oxygen protecting group is referred to as an "oxygen deprotecting agent". The oxygen deprotecting agent is selected independently for each occurrence. When removing the oxygen protecting group, a cation scavenger is generally present. Examples of suitable cation scavengers include triisopropylsilane (TIS) and DTT. The cation scavenger is selected independently for each occurrence. A preferred cation scavenger is triisopropylsilane.

[0143] Cleavage of the compounds disclosed herein from a solid support (e.g., Wang resin) can be achieved using TFA in water in the presence of DTT. In one preferred embodiment, when the compound is cleaved from the solid support, all tBu groups are removed. This can be achieved using a TFA cleavage cocktail (3 mL; 95% TFA, 5% H2O, 25 mg / mL DTT).

[0144] All solvents disclosed herein may be used, if desired, in combinations of two or more solvents.

[0145] The term "solid support" refers to a resin used in solid-phase synthesis. Examples include Wang resin, which is polystyrene cross-linked with divinylbenzene.

[0146] A related aspect provides a compound of formula A-1 below having a chemical purity of greater than 90%, greater than 95%, greater than 98%, or greater than 99% and containing 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% or less of one or more structural impurities (including any of the structural impurities of formulae B-1, B-1-R, B-1-S, B-2, B-2-R, B-2-S, B-3, B-3-R, and B-3-S above).

[0147]

Chemical formula

[0148] The compound of Formula A-1 is also referred to herein as PSMA-62.

[0149] In another aspect, a radiopharmaceutical agent is provided that is a complex of a compound of Formula A-1 and one or more radioisotope atoms. In some embodiments, the agent has a purity or chiral purity of greater than 90%, greater than 95%, greater than 98%, or greater than 99%.

[0150] Exemplary radioisotopes include 47 Sc, 51 Cr, 57 Co, 58 Co, 59 Fe, 67 Cu, 67 Ga, 68 Ga, 71 Ge, 72 As, 72 Se, 77 As, 77 Br, 81 Rb, 88 Y, 90 Y, 97 Ru, 99 mTc, 100 Pd, 101 mRh, 103 Pb, 105 Rh, 109 Pd, 111 Ag, 111 In, 113 In, 119 Sb, 121 Sn, 127 Cs, 128 Ba, 129 Cs, 131 Cs, 131 I, 139 La, 140 La, 142 Pr, 143 Pr, 149 Pm, 151 Eu, 153 Eu, 153 Sm, 159 Gr, 161 Tb, 165 Dy, 166 Ho, 169 Eu,175 Yb, 177 Lu, 186 Re, 188 Re, 189 Re, 191 Os, 193 Pt, 194 Ir, 197 Hg, 198 Au, 199 Ag, 199 Au, 201 Tl, 203 Pb, 211 At, 212 Bi, 212 Pb, 213 Bi, 225 Ac, and 227 Th are included. In certain embodiments, the radioisotope is 67 Cu, 177 Lu, 212 Bi, 161 Tb, 212 Pb, 223 Ra, or 225 Ac. In certain embodiments, the radioisotope is 177 Lu, 225 Ac, 211 At, 67 Cu, 161 Tb, 67 Ga, 68 Ga, 203 Pb, 223 Ra, or 212 Pb. In certain embodiments, the radioisotope is 225 Ac. In certain embodiments, the radioisotope is 68 Ga. In certain embodiments, the radioisotope is 177 Lu.

[0151] The radiopharmaceuticals of the present invention, namely, the compounds of any of formulas A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S, can be readily prepared. Preferred syntheses are described in the following examples and can provide high-purity compounds at low levels even in the presence of structural impurities as disclosed herein.

[0152] Radioactive label. Typically in a solvent or solvent mixture, as a result, the radioisotope compound is heated and stirred for a sufficient time in the compounds of A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S in which the radioisotope is not pre-chelated, so that a compound of the above formula complexed or chelated with the radioisotope compound can be provided.

[0153] In particular, for the preparation of the compound of formula A-3-R ( 225 Ac-PSMA-62), the following protocol can be utilized: Ac-225 (for example, using the reagent of ( 225 Ac)Cl3) is prepared in 0.1 M HCl, added to a vial, followed by a reaction buffer, 1 M NaOH for pH adjustment, and finally the PSMA-62 precursor (a compound of formula A-3-R in which Ac-225 is not complexed). The reaction buffer can suitably be prepared by dissolving 1878 mg of sodium ascorbate and 924 mg of ammonium acetate in 10 mL of sterile water for injection (SWFI). Other formulations can suitably be effective reaction buffers. Preferred reaction buffers can contain one or more ascorbate compounds. The prepared reaction mixture contains Ac-225 at 10 MBq / mL, sodium ascorbate (32 mg / mL), ascorbic acid (0.73 mg / mL), PSMA-62 (214 μg / mL) at pH 5.5. The reaction mixture is heated at a time and temperature sufficient for Ac-225 to effectively complex with the PSMA-62 molecules and with stirring (for example, while shaking the reaction vessel) to achieve an uptake of radioisotope exceeding, for example, 90, 95, or 98%. In one protocol, the reaction mixture is stirred, heated to 90 °C for 30 minutes, then diluted with a formulation buffer containing 0.1 mg / mL of DTPA in PBS to a final pH of 6.5, and the product ( 225 Ac-PSMA-62) RAC is set to 1 MBq / mL.

[0154] 225 Ac can be produced using methods known in the art. 225 The two most important production routes for the production of Ac are233 Obtained from waste stockpiles containing U 229 Separation from the natural decay of Th, and by high-energy protons (above 70 MeV) 232 Irradiation of Th 225 As a result of the method for preparing Ac, thorium can be an impurity in the chelating compound. More specifically 229 Th and 232 At least one of Th 225 Can be an impurity in Ac

[0155] 225 Another way to form Ac involves the irradiation of Ra-226 using intermediate-energy protons and photons

[0156] Exemplary preferred radiolabeling protocols for providing compounds of formula A-3-R (225Ac-PSMA-62) are also shown in Example 4 below. Exemplary preferred radiolabeling protocols for providing compounds of formula B-1-R ( 68 Ga-PSMA-62) are described in Example 9 below

[0157] As considered, pharmaceutical compositions (also referred to as formulations or pharmaceutical formulations) of any of the compounds of formulae A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S above and one or more pharmaceutically acceptable carriers are provided. In some embodiments, the pharmaceutical composition contains no radiochemical impurities in an amount of 5, 4, 3.5, 3, 2.5, 2, 1.5, 1 or 0.5 wt% or less based on the total weight of the pharmaceutical composition, as may be determined by radiometric detection (including HPLC radiometric detection), the composition is maintained, for example, at 2 to 8 °C or below, and such purity levels are shown for 3, 4 or 5 days or longer after preparation of the composition

[0158] In some embodiments, the pharmaceutical composition is free of chemical impurities in an amount of 5, 4, 3, 2, 1 or 0.5% by weight or less based on the total weight of the pharmaceutical composition, as determined by, for example, HPLC, ultra-high performance liquid chromatography (UHPLC), or chromatography including HPLC / UV analysis, or other methods, the composition is maintained at 30 degrees or less, and such purity levels are demonstrated for 3, 4 or 5 days or longer after preparation of the composition.

[0159] In another aspect, there is provided a pharmaceutical composition comprising a radiopharmaceutical agent disclosed herein and one or more pharmaceutically acceptable carriers.

[0160] Treatment methods As described above, methods and uses for diagnosing and / or monitoring disorders including cancer disclosed herein are provided. In one aspect, these methods and uses may include administering to a subject, such as a human patient, a radiocontrast compound comprising a PSMA targeting agent. Examples of PSMA targeting agents include, for example, 68 Ga-PSMA-11, 18 F-DCFPyl, or a radiopharmaceutical agent disclosed herein, particularly 68 agents complexed with Ga. In one aspect, 68 Ga-PSMA-62 is a preferred radiocontrast agent. The protocol can include 68 allowing a time for a radiopharmaceutical such as Ga-PSMA-11, 18 F-DCFPyl, or 68 Ga-PSMA-62 to bind to the tissue of the subject (e.g., at least or up to 5, 10, 15, 20, 30, 40, 50, or 60 minutes), and then monitoring the subject (outside the body), particularly by other systems such as positron emission tomography (PET) or single photon emission computed tomography (SPECT). Then, based on the evaluation from the ex vivo monitoring, the patient may be further treated, particularly, 225 selected for treatment with one or more radiopharmaceutical compounds disclosed herein including Ac PSMA-62.

[0161] As examined, the use of radiopharmaceuticals (i.e., compounds of any of formula A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S) is provided for treating cancers including biochemical recurrence (BCR) prostate cancer, non-metastatic prostate cancer, and metastatic prostate cancer, including prostate cancer such as and / or hormone-sensitive prostate cancer, castration-resistant prostate cancer (CRPC), and anti-androgen drug (e.g., enzalutamide)-resistant prostate cancer, and / or oligometastatic hormone-sensitive prostate cancer (OmHSPC).

[0162] In such a method, the radiopharmaceutical can be administered to a subject such as a human in an amount effective (e.g., reduction in tumor size) for treating cancer, such as in a dose of about 0.001 MBq to about 100 MBq, 0.01 MBq to about MBq, or 0.1 MBq to about 100 MBq, and can be preferably administered as a bulk solution in a vial or syringe prepared from a unit dose in a vial or syringe, or in a local or central pharmacy handling the radiopharmaceutical or through a cGMP central manufacturing facility, or from a cold kit prepared using lutetium-177. For the compounds disclosed herein containing the radioisotope Ac-225, a dose within the range of 0.1 to 100 MBq / mL may be preferred. In some embodiments, for the radioactivity concentration (RAC), non-limiting examples include 0.001 MBq / mL to 100 MBq / mL, 0.01 MBq / mL to 50 MBq / mL, and 0.01 MBq / mL to 33 MBq / mL. In some embodiments, the dose size ranges from about 1 to 20 or 3 to 10 mL. For the compounds disclosed herein containing radioisotopes other than Ac-177 such as Lu-225, higher doses, e.g., doses of 10 GBq or 20 GBq or more, may be appropriate.

[0163] In certain embodiments, the dose may be administered as described in the following table.

[0164]

Table 1

[0165] In certain embodiments, the subject has prostate cancer, such as castration-sensitive prostate cancer, castration-resistant prostate cancer, metastatic castration-resistant prostate cancer, advanced prostate cancer, anti-androgen resistant prostate cancer (e.g., enzalutamide-resistant prostate cancer, abiraterone-resistant prostate cancer, bicalutamide-resistant prostate cancer), drug-resistant prostate cancer such as AR-V7-induced drug-resistant prostate cancer (e.g., AR-V7-induced enzalutamide-resistant prostate cancer), AKR1C3-induced drug-resistant prostate cancer (e.g., AKR1C3-induced enzalutamide-resistant prostate cancer), oligometastatic hormone-sensitive prostate cancer, and combinations thereof, such as one or more of the foregoing.

[0166] In a further particular embodiment, the subject is a human suffering from oligometastatic hormone-sensitive prostate cancer, and an effective amount of the radiopharmaceutical agent disclosed herein is administered to the subject for treating prostate cancer.

[0167] In a further particular aspect, the subject is a human suffering from metastatic castration-resistant prostate cancer, and an effective amount of a radiopharmaceutical agent is administered to the subject for treating prostate cancer.

[0168] The effective amount of the radiopharmaceutical agent administered to a patient is generally determined by considering the patient's record. However, the effective amount may preferably be in the range of about 0.1 GBq to 100 GBq per dose, for example when the radiopharmaceutical agent contains Lu-177. Further, a dose selected from 1.5 MBq to 13.8 MBq can be chosen. For example, 1.5 MBq. For example, 2.3 MBq. For example, 3.0 MBq. For example, 4.6 MBq. For example, 6 MBq. For example, 9 MBq. For example, 9.2 MBq. For example, 13.8 MBq. For the compounds disclosed herein containing the radioisotope of Ac-225, a dosage within the range of 0.1 to 100 MBq / mL may be preferred. This dosage can be appropriately administered as a unit dose in a vial or syringe, or as a bulk solution in a vial or syringe prepared from a cold kit at a local or central pharmacy handling radiopharmaceuticals or through cGMP central manufacturing.

[0169] If necessary or desirable, the treatment may involve more than one administration of an effective amount of the radiopharmaceutical agent.

[0170] The compound may also contain at least one non-radioactive, non-toxic carrier metal. For example, the carrier metal can be selected from Bi and Fe. For example, the non-radioactive carrier metal may enable MRI imaging (e.g., Fe) or X-ray contrast imaging (e.g., Bi). A further example of a carrier metal is trivalent bismuth, which further provides X-ray contrast in microspheres, such that the microspheres can be imaged in CT.

[0171] The compounds of the present invention may also be utilized as a contrast agent, for example, in image-guided surgery. For example, the compounds disclosed herein can be administered to a subject in an amount sufficient to preferentially localize to the target tissue of the surgery, and the surgeon can detect the presence or absence of the contrast agent during the surgical procedure.

[0172] Combination Therapy The radiopharmaceuticals disclosed herein may preferably be administered to a subject in combination with or in conjunction with one or more other therapeutic agents, particularly one or more other chemotherapeutic agents.

[0173] In one aspect, a subject may receive treatment with a radiopharmaceutical disclosed herein in combination with a regimen that may include chemotherapy, such as docetaxel; cisplatin; gemcitabine; cisplatin / gemcitabine; cabazitaxel; mitoxantrone; estramustine phosphate; one or more antiandrogen agents, such as one or more LHRH agonists, such as leuprolide and goserelin, or antagonists (e.g., flutamide and relugolix); one or more antiandrogen agents, such as flutamide, nilutamide, bicalutamide, cyproterone, abiraterone, enzalutamide, darolutamide, and apalutamide; particularly for subjects suffering from prostate cancer, including metastatic castration-resistant prostate cancer, one or more PARP inhibitors such as rucaparib or niraparib.

[0174] In a further aspect, a subject may receive treatment with a radiopharmaceutical disclosed herein in combination with an immunotherapy regimen that may include adoptive cell therapy or adoptive immunotherapy.

[0175] For example, to treat a patient suffering from cancer, including to treat a disease associated with the expression of a cancer or tumor antigen, the radiopharmaceuticals disclosed herein may be administered in combination with (e.g., in CAR T cell therapy) immune effector cells (e.g., T cells, NK cells) engineered to express a chimeric antigen receptor.

[0176] For patients suffering from cancers including prostate cancer, the radiopharmaceuticals disclosed herein may also be administered in combination with other immune-boosting approaches including other immune-based therapies or antibody therapies such as sipuleucel-T (Provenge). For example, in one protocol, to treat patients suffering from cancer, particularly prostate cancer, the radiopharmaceuticals disclosed herein may be administered in combination with one or more monoclonal antibodies such as pembrolizumab (Keytruda), ipilimumab (Yervoy), and / or nivolumab (Opdivo).

[0177] As used herein, the term "in combination" in the context of administering therapies to a subject refers to using two or more therapies for a therapeutic benefit. The term "in combination" in the context of administration can also refer to the prophylactic use of a therapy to a subject when used with at least one additional therapy. The use of the term "in combination" does not limit the order in which the therapies (e.g., first and second therapies) are administered to the subject. The therapies can be administered before (e.g., 1 minute, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), simultaneously with, or after (e.g., 1 minute, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of a second therapy to a subject in need of the treatment disclosed herein. The therapies are administered to the subject within an order and time interval such that the therapies can act together. In certain embodiments, the therapies are administered to the subject within an order and time interval that provides an increased benefit compared to when administered within other orders and time intervals. Any additional therapies can be administered in any order with other additional therapies.

[0178] Packaged Radiopharmaceuticals and Kits As discussed above, provided are also therapeutic kits, including cold kits, in which the radiopharmaceuticals disclosed herein can be prepared immediately prior to administration in a medical facility, such as a hospital laboratory or a pharmacy that handles radiopharmaceuticals. In such kits, the compound may be provided in a vial or other container in a lyophilized or other form separated from a radioisotope such as Ac-225, Lu-177, or other radioisotopes. The compound and the radioisotope can be reacted in a medical facility to provide a radiopharmaceutical of formula A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S, which can then be rapidly administered to a patient.

[0179] In a further aspect, provided are also packaged preparations or products of radiopharmaceuticals. The packaged preparation can include 1) a radiopharmaceutical, such as a compound of formula A-2, A-2-R, A-2-S, A-3, A-3-R, and / or A-3-S that forms a complex with a radioisotope, and 2) instructions for using the radiopharmaceutical for treating cancer, such as prostate cancer. Preferably, the packaged preparation includes a therapeutically effective amount of the radiopharmaceutical.

[0180] In certain exemplary packaged preparations or products, the radiopharmaceutical can be suitably packaged in a suitable container, such as one labeled for use as a therapy for treating a subject suffering from, for example, prostate cancer. The container can contain the radiopharmaceutical in an aqueous formulation, optionally together with one or more stabilizers or buffers. The product can include a container (such as a vial) containing the radiopharmaceutical. Additionally, the manufactured product or kit may further include, for example, packaging materials, instructions for use, syringes, delivery devices for treating a targeted condition such as prostate cancer or other cancers.

[0181] The packaged system or product may also include instructions (e.g., a printed label or attached document or other medium (e.g., an audio or video file) that describes the use of the product). The instructions can be associated with (e.g., affixed to) the container and can describe the manner in which the composition therein should be administered (e.g., the frequency and route of administration), the instructions therefor, and other uses discussed above. The composition may be immediately administrable (e.g., the dosage is present in appropriate units) but may also contain one or more pharmaceutically acceptable adjuvants, carriers, or other diluents as additional pharmaceuticals.

[0182] In certain embodiments, 225 The radiopharmaceuticals disclosed herein that contain Ac PSMA-62 may be stored and supplied in a suitable container, particularly a glass container such as a vial, e.g., a type I glass vial capped with a septum and an aluminum crimp, as discussed. 225 The Ac PSMA-62 composition is suitably formulated as an aqueous formulation containing one or more excipients such as one or more of ammonium acetate, sodium ascorbate, hydrochloric acid, and diethylenetriaminepentaacetic acid (DTPA), and the excipients may be present at concentrations commonly used in IV radiopharmaceuticals. The radiopharmaceutical can be suitably transported in a container to maintain a temperature of 2-8°C. The container may preferably be lead-shielded.

[0183] In the clinical setting, the radiopharmaceutical is preferably maintained at a low temperature prior to use, e.g., the radiopharmaceutical may be at 2-8°C in a lead pot at the clinical site. Preferably, the radiopharmaceutical 225 is administered to the patient within 5 days, including within 4, 3, or 2 days after the preparation of a radiopharmaceutical such as Ac PSMA-62.

[0184] In certain embodiments, 225 the dosing of the radiopharmaceuticals disclosed herein, such as Ac PSMA-62, to a patient may be based on the patient's body surface area (BSA). The dosage based on the patient's body surface area (BSA) is calculated using the DuBois formula (BSA = 0.007184 *Height 0.725* Weight 0.425 ) can be appropriately calculated using (DuBois and DuBois, 1989, Nutrition, 5:303-11). 225 The total administered radioactivity of 225Ac PSMA-62 can appropriately be 1.5 - 13.8 MBq (±10%) for patients with mCRPC at 4 cycles every 6 weeks or for patients with BCR prostate cancer or OmHSPC at 2 cycles every 8 weeks. Preferred dosages can also include administering 225Ac PSMA-62 one or more times (e.g., 2, 3, or 4 times) on the first day during a treatment cycle (e.g., a 2-week cycle, 3-week cycle, 4-week cycle, 5-week cycle, 6-week cycle, 7-week cycle, 8-week cycle, 9-week cycle, 10-week cycle, 11-week cycle, or 12-week cycle). Further, the dosage can be selected from 1.5 MBq to 13.8 MBq. For example, 1.5 MBq. For example, 2.3 MBq. For example, 3.0 MBq. For example, 4.6 MBq. For example, 6 MBq. For example, 9 MBq. For example, 9.2 MBq. For example, 13.8 MBq. 225 Radiopharmaceuticals disclosed herein, such as 225Ac PSMA-62 injection, are appropriately administered intravenously in a volume up to 10 mL. The dosage is preferably not diluted prior to intravenous administration and is preferably administered intravenously into an upper limb vein, preferably the antecubital vein. The patient can be encouraged to drink fluids regularly and urinate on the day of administration both before and after administration.

[0185] All references cited herein are incorporated herein by reference in their entirety. The following non-limiting examples are illustrative.

Example

[0186] Example 1 General information: Fmoc - amino acid analogs were purchased from Chem-Impex, Asta Tech or Iris Biotech: Fmoc-Asp-D-Oall (Asta Tech, catalog number F11941), Fmoc-D-Orn(Dde)-OH (Chem-Impex, catalog number 30521), Fmoc-D-Lys(OtBu)-OH (Iris Biotech, catalog number FAA4690), Fmoc-D-Ala(2-naph)-OH (Asta Tech, catalog number F12304), Fmoc-D-Tyr(OtBu)-OH (Chem-Impex, catalog number 02465), Fmoc-Phe(4-NHBoc)-OH (Chem-Impex, catalog number 03755). 2-Chlorotrityl chloride (2-CTC) resin was purchased from Chem-Impex (catalog number 12996, initial loading: 1.43 mmol / g). All necessary solvents and other organic reagents were purchased from Sigma-Aldrich. Solid-phase synthesis of peptides was performed manually using an IKA KS 130 Basic shaker. Preparative RP-HPLC of peptides was achieved using a Varian semi-preparative system equipped with a Discovery C18 569226-U RP-HPLC column. The mobile phase for semi-preparative HPLC was typically prepared by mixing water (0.1% TFA) with acetonitrile in a gradient concentration. Mass spectra and HPLC retention times were recorded using a Hewlett Packard HPLC / MSD system equipped with a UV detector (monitoring at 215 nm) using a ZORBAX Eclipse Plus C18 RP-HPLC column (4.6×50 mm, 1.8 μm), or using an Agilent 300SB-C8 RP-HPLC column (4.6×100 mm, 3.5 μm) with a solvent gradient of A) water (0.1% TFA) and B) acetonitrile at a flow rate of 0.5 mL / min on a Thermo LTQXL LC / MS system equipped with UV detectors (monitoring at 215 nm, 254 nm). Unless otherwise stated, all HPLC retention times were given for an eluent gradient of 2% B for the first 3 minutes, then 2% - 98% B over 6 minutes, which was maintained for the next 6 minutes. D-Orn is (2R)-2,5-diaminopentanoic acid.

[0187] (1) Synthesis Protocol (SP) SP-1: 2-CTC Resin Loading: Load Fmoc-AA-OH (1.5 equivalents) in anhydrous dichloromethane (DCM) containing N,N-diisopropylethylamine (DIPEA) (4.5 equivalents) onto 2-CTC resin (1.43 mmol / g) at room temperature (RT) for 2 hours. Cap the remaining trityl chloride by adding 2 mL / g of methanol (MeOH) for 15 minutes. Then, filter the resin and wash it thoroughly with DCM (2x), dimethylformamide (DMF) (2x), and MeOH (2x) respectively, and store it under vacuum overnight. Loading is determined using the weight difference.

[0188] SP-2: Peptide Synthesis by TBTU / HOBt Coupling: Add a solution of Fmoc-AA-OH (2.0 equivalents), N,N,N’,N’-tetramethyl-0-(benzotriazol-1-yl)uronium tetrafluoroborate (TBTU) (2.0 equivalents), N-hydroxybenzotriazole (HOBt) (2.0 equivalents), DIPEA (4.5 equivalents) in DMF (8 mL / g resin) to the resin-bound free amine peptide, shake it at room temperature for 2 hours, and wash it with DMF (6x). After completion of the reaction, wash the resin with DMF (6x).

[0189] SP-3: Fmoc Deprotection on Resin: Treat the resin-bound Fmoc-protected peptide with 20% piperidine (v / v) in DMF for 5 minutes, and the second time for 15 minutes. Then, wash the resin thoroughly with DMF (8x).

[0190] SP-4: Dde Deprotection on Resin: The N-(1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-ethyl) (Dde)-protected peptide (1.0 equivalent) was dissolved in a solution of 2.0% hydrazine monohydrate (N2H4·H2O) in DMF (v / v). After 15 minutes, the resin-loaded peptide was washed with DMF (6x). When both Fmoc and Dde protecting groups are present and only Dde deprotection is required, the resin-loaded peptide was treated with a solution containing NH2OH·HCl (630 mg), imidazole (460 mg), DCM (0.5 mL), DMF (0.5 mL), and N-methyl-2-pyrrolidone (NMP) (2.5 mL) at room temperature for 3 hours. Then, the resin-loaded peptide was washed with DMF (6x).

[0191] SP-5: Allyl Ester Deprotection on Resin: The O-allyl protecting group was removed from the resin-bound peptide using a solution of DCM (6.0 mL) containing triisopropylsilane (TIPS) (50.0 equivalents) and tetrakis(triphenylphosphine)palladium(0) (Pd(PPh3)4) (0.3 equivalent). The resin was treated with this solution at room temperature for 1.5 hours. Finally, the resin was washed with DCM (3x) to remove Pd(PPh3)4.

[0192] SP-6.1: A) Peptide Cleavage from Resin Retaining Side Chain Protecting Groups. The fully protected resin-bound peptide was dissolved in 20 DCM / trifluoroethanol (TFE) / acetic acid (AcOH) (6 / 3 / 1; v / v / v) and shaken for 30 minutes. The solution was filtered off, and the resin was dissolved in another cleavage solution for an additional 30 minutes. The fractions were combined, the solvent was concentrated in vacuo, the residue was purified by semi-preparative HPLC, and then lyophilized to obtain the pure protected peptide.

[0193] SP-6.2: B) Peptide Cleavage from Resin with Simultaneous Deprotection of All Acid-Labile Protecting Groups. The fully protected resin-bound peptide was dissolved in a mixture of TFA / TIPS / water (4:4:1; v / v / v) and shaken for 2 hours. The filtrate was concentrated in vacuo, the residue was purified by semi-preparative HPLC, and then lyophilized to obtain the pure peptide.

[0194] (2) Synthesis of (R)-tert-Bu4-DOTAGA. (R)-tert-Bu4-DOTAGA is either commercially available or can be readily prepared according to chemical schemes known in the art.

[0195] (3) Synthesis of the structural unit I-7a (Dde-D-Asp-[Fmoc-Phe(4-NHBoc)-D-Tyr(OtBu)-D-Ala(2-naph)-D-Lys-OtBu]): Synthesis of the exemplary compound of I-7a (Dde-D-Asp-[Fmoc-Phe(4-NHBoc)-D-Tyr(OtBu)-D-Ala(2-naph)-D-Lys-OtBu]): Fmoc-D-Lys-OtBu / PyBroP / DCM / DIPEA was used. Only a non-significant amount (less than 0.5%) of racemization / isomerization was observed by chiral HPLC.

[0196] Synthesis scheme of the intermediate I-7a.

[0197]

Chemical Structure

[0198] The Asp side-chain carboxylic acid was conjugated using 2-Cl-Trt-resin. This bulky resin can prevent racemization and isomerization that may occur at the Asp moiety during the coupling reaction. Further, even if some aspartimide is formed, considering that the mechanism of aspartimide formation cleaves the resin-amino acid bond or the peptide bond, the aspartimide can be washed away from the resin. Further, here PyBroP + DIPEA in DCM was used for the coupling reaction. Compared with "TBTU + HOBt + DIPEA in DMF", the condition of "PyBroP + DIPEA in DCM" can give a much cleaner product and greatly reduce racemization and isomerization.

[0199] The first resin loading with Fmoc-D-Asp-OAll was carried out as described in SP-1. The amino group was Fmoc-deprotected according to SP-3 and re-protected at room temperature using Dde-OH (2.0 eq) and DIPEA (4.0 eq) in DMF. The reaction mixture was shaken overnight. Then, applying SP-5 achieved the allyl deprotection of O-Asp, and the free carboxylic acid of Asp was coupled with Fmoc-D-Lys-OtBu·HCl according to SP-2, except that 2.0 eq (instead of 1.5 eq) of Fmoc-D-Lys-OtBu·HCl, 2.0 eq of PyBroP (Aldrich, catalog number 18565) (instead of TBTU / HOBt), and 4.0 eq of DIPEA in DCM (instead of DMF) were used. After removing Fmoc according to SP-3, the next steps involved iterative conjugation with Fmoc-D-Ala(2-naph)-OH, Fmoc-D-Tyr(OtBu)-OH, and Fmoc-L-Phe(4-NHBoc)-OH according to SP-2. Finally, the peptide was cleaved from the resin according to SP-6.1 (method A), and the crude product was purified by semi-preparative RP-HPLC to obtain pure I-11a (total yield 60% for loading Fmoc-D-Asp(2-ClTrt-resin)-Oall at 1 mmol scale) as a white powder. LC-MS (ESI+) m / z (relative intensity): 1382.5 ([M+H]+, 100); tr = 11.06 min.

[0200] Conditions: The mass spectrum and HPLC retention time were recorded on a Thermo LTQXL LC / MS system equipped with an Agilent 300SB-C8 RP-HPLC column (4.6×100 mm, 3.5 μm), using a solvent gradient of A) water (0.1% TFA) and B) acetonitrile at a flow rate of 0.5 mL / min and monitoring with a UV detector (at 215 nm and 254 nm). The eluent gradient was 5% B for the first 3 minutes, then 5% - 98% B over 6 minutes, which was maintained for the next 5 minutes (0 - 3 min: 5% B; 3 - 9 min: 5 - 98% B; 9 - 15 min: 98% B).

[0201] (4) Synthesis of the structural unit I-11a. To a solution of L-glutamic acid di-tert-butyl hydrochloride (AstaTech, catalog number 34103, 3.9 g, 13.2 mmol) in dichloromethane (150 mL) was added pyridine (3.0 mL). The mixture was cooled to -10 °C, and a solution of triphosgene (2.58 g, 9 mmol, 0.67 equiv) in dichloromethane (15 mL) was added thereto. The mixture was stirred at -10 °C for 2 h under an argon atmosphere, then 0.1 M aqueous HCl was added, and the whole was extracted with dichloromethane. The organic solution was washed with saturated brine, dried over sodium sulfate, and evaporated under reduced pressure to obtain 3.8 g of ditBuGlu-NCO, which was redissolved in dichloromethane (150 mL). H-Glu(Obzl)-OtBu.HCl (AstaTech, catalog number 34691, 4.5 g) and DIPEA (10 mL) were added. The mixture was stirred at room temperature overnight, then 0.1 M aqueous HCl (150 mL) was added. The organic phase was separated, washed with aqueous NaHCO3 and saturated brine, dried over sodium sulfate, and evaporated. The residue was purified by silica gel flash column chromatography (n-hexane / ethyl acetate) to obtain the Bzl-protected compound, which was added to a suspension of 10% Pd-C (1.0 g) in methanol (150 mL). The mixture was degassed under reduced pressure and placed under H2 (50 psi). The mixture was stirred at room temperature for 2 h until the reaction was complete. Then the catalyst was removed by filtration through celite. The filtrate was concentrated in vacuo until completely dry to obtain the compound as a white powder (5.8 g, 90%). LC-MS (ESI+) m / z (relative intensity): 488.8 ([M+H]+, 100); tr = 10.3 min.

[0202] Conditions: The mass spectrum and HPLC retention time were recorded using a Hewlett Packard HPLC / MSD system equipped with a UV detector (monitoring at 215 / 254 nm) and a ZORBAX Eclipse Plus C18 RP-HPLC column (4.6×50 mm, 1.8 μm), with solvent gradients of A) water (0.1% TFA) and B) acetonitrile used at a flow rate of 0.5 mL / min. The eluent gradient was 2% B for the first 3 minutes, then increased from 2% to 98% B over 6 minutes and maintained at 98% B for the next 5 minutes (0 - 3 min: 2% B; 3 - 9 min: 2 - 98% B; 9 - 15 min: 98% B).

[0203] (5) Synthesis of Compound A-1. In the following convergent route, intermediate building block I-7a was used. On the other hand, PyBroP / DCM / DIPEA was also used to minimize potential racemization and isomerization. After a standard preparative HPLC purification process, the pure target compound was obtained. Only a non-significant amount (less than 0.5%) of racemization / isomerization was observed.

[0204] Synthesis of Exemplary Compounds of Formula A-1

[0205] [Chemical formula]

[0206] The first resin loading with Fmoc-D-Orn(Dde)-OH was carried out as described in SP-1. After Fmoc-deprotection according to SP-3, the building block I-11a (1.5 eq) was coupled to D-Orn(Dde) according to SP-2. In the next step, the Dde protecting group was cleaved according to SP-4, and the free amino group was treated with the building block I-7a (Dde-D-Asp-[Fmoc-Phe(4-NHBoc)-D-Tyr(OtBu)-D-Ala(2-naph)-D-Lys-OtBu]) according to SP-2, except that 2.0 eq (instead of 1.5 eq) of the building block I-7a, 2.0 eq of PyBroP (Aldrich, catalog number 18565) (instead of TBTU / HOBt) and 4.0 eq of DIPEA in DCM (instead of DMF) were used. After removing Fmoc with N-terminal Fmoc-L-Phe(4-NHBoc)-OH according to SP-3, the free amino group was coupled to (R)-tert-Bu4-DOTAGA using 2.0 eq (instead of 1.5 eq) of (R)-tert-Bu4-DOTAGA, 2.0 eq of PyBOP (instead of TBTU / HOBt) and 4.5 eq of DIPEA in 5:1 (v / v) DCM / DMF (instead of DMF) according to SP-2. To conjugate TMA to D-Asp, SP-4 was applied to achieve Dde deprotection and obtain the free amino group. TMA was coupled using TMA (2.0 eq), HOBt (1.5 eq), TBTU (1.5 eq) and DIPEA (10 eq) in DMF. The reaction was shaken at room temperature for 8 h. After conjugation of TMA, the peptide was cleaved from the resin using a mixture of TFA / TIPS / water (50:3:3). The reaction was shaken at room temperature for 48 h and then filtered off. The filtrate was concentrated in vacuo and the residue was purified by semi-preparative RP-HPLC to give the pure compound (13% overall yield based on loading of Fmoc-D-Orn(Dde)-2-ClTrt-resin on a 0.1 mmol scale) as a white powder. LC-MS (ESI+) m / z (relative intensity): 1852.6: ([M+H]+, 17), 1234.8 (20), 926.4 (100), 618.1 (35); tr = 16.16 min.TMA is 3,5-bis(tert-butoxycarbonyl)benzoic acid.

[0207] Conditions: Mass spectra and HPLC retention times were recorded using an Agilent 1290 HPLC / 6460 Triple Quad LC / MS system equipped with a UV detector (monitoring at 215 nm and 254 nm) using an XBridge BEH Phenyl Column (130A, 4.6×150 mm, 3.5 μm) with solvent gradients of A) water (0.1% TFA in HPLC-grade water, pH 2.02) and B) acetonitrile (0.08% TFA in ACN), starting at 85% A and 15% B and increasing to 76% A and 24% B at a rate of 1.0 mL / min over 20 minutes. Then, it was increased to 64% A and 36% B over 4 minutes. Finally, the gradient parameters were returned to the initial starting conditions (0 - 20 min: 15 - 24% B; 20 - 24 min: 24 - 36% B). Data was analyzed using Chemstation software from Agilent.

[0208] Example 2 As shown in the figure, a compound of formula A-1 was prepared using a linear solid-phase synthesis scheme. This compound was obtained with a chiral purity of over 98%.

[0209] Specifically, as shown in Figure 2, after the preparation of Fmoc-D-Lys(Mtt)-Wang resin (0.50 meq / g loading), the coupling of D-2-naphthylalanine via 4-(Boc-amino)-phenylalanine was carried out using DIC / oxyma in DMF. All subsequent couplings from (R)-DOTA-GA(tBu)4 were carried out using PyAOP / DIEA. Table 1 below shows the synthetic steps. Mtt is 4-methyltrityl.

[0210] After the coupling of (R)-DOTA-GA(tBu)4, the D-Lys(Mtt) side chain was deprotected with HFIP / DCM. Subsequently, Fmoc-D-Asp(OAll)-OH was coupled to the D-lysine side chain using PyAOP / DIEA as the activator, followed by di-tBu ester of 1,3,5-benzenetricarboxylic acid. The removal of the allyl ester protection of the D-Asp side chain was carried out using Pd(PPh3)4 / 1,3-dimethylbarbituric acid (DMBA) in DMF / DCM. The coupling of Fmoc-D-Orn-OtBu, followed by HO-γGlu(OtBu)-CO-Glu(OtBu)-OtBu, was completed using PyAOP / DIEA to complete the sequence. HFIP is hexafluoro-2-propanol.

[0211] Table 1. Synthesis steps [Table 2]

[0212] Pd removal was carried out by treatment with a solution of DEDTC / DMF.

[0213] TFA cleavage and precipitation. Cleavage of the compound from the resin and overall deprotection were carried out using TFA / H2O / TIS for 15 hours.

[0214] The TFA cleavage solution was concentrated, cooled in a dry ice bath, and cold MTBE (-78 °C) was slowly added thereto. The solution was homogenized and additional MTBE was added. The resulting precipitate was filtered to obtain a crude peptide purity of over 75%.

[0215] Table 2. Summary of crude yields [Table 3]

[0216] HPLC purification. Purification of the crude peptide was carried out on a NovaSep column packed with Daisogel-SP-120-10-ODS-RPS C18 medium. The column was equilibrated under starting gradient conditions.

[0217] The purification in step 1 was carried out on the crude product dissolved in NH4HCO3 / H2O. The product was eluted with a gradient of 2 - 20% CAN in 0.1 M NH4HCO3 / H2O, and the eluate was monitored by UV at 254 nm. The product eluted at approximately 6% CAN. All fractions were acidified with pure TFA, and the fractions containing the product with a purity of 94% or higher were pooled and subjected to RP - HPLC step 2.

[0218] The purification in step 2 was carried out with a gradient of 5 - 25% CAN in 0.1% TFA / H2O. A product eluting between 17 - 20% CAN was observed. The fractions containing the product with a purity of 95% or higher were pooled, filtered, shell - frozen, and lyophilized to produce the product in the form of a dry powder as the TFA salt.

[0219] In total, 10.1 g of the second lot of the crude product was subjected to purification to produce 4.60 g of the purified substance. The purity was 97.4%. Theoretical yield for 11.50 mmol of synthesis: 21.29 g. Theoretical yield for 10.09 g of 23.457 g of the crude product: 9.16 g. % Yield: 50.2%.

[0220] Example 3: Purity analysis of PSMA - 62 produced by convergent synthesis. The PSMA - 62 compound (formula A - 3 - R) produced by the method of Example 1 above was evaluated as follows. Purity by HPLC: Conditions: The mass spectrum and HPLC retention time were recorded using an Agilent 1290 HPLC / 6460 Triple Quad LC / MS system equipped with a UV detector (monitoring at 215 nm and 254 nm) and an Xbridge BEH Phenyl Column (130A, 4.6×150 mm, 3.5 μm). Solvent gradient A) water (0.1% TFA in HPLC grade water, pH 2.02) and B) acetonitrile (0.08% TFA in CAN) were used, starting at 85% A and 15% B and recording at a flow rate of 1.0 mL / min, increasing to 76% A and 24% B over 20 minutes; then increasing to 64% A and 36% B over 4 minutes. Finally, the gradient parameters were returned to the initial starting conditions (0 - 20 minutes: 15 - 24% B; 20 - 24 minutes: 24 - 36% B). Data was analyzed using Chemstation software from Agilent.

[0221] LC / MS spectrum calculated MW: 1851. A peak at 16.11 minutes was observed, 1852.4: ([M + H]+, 15), 1235.1 (2 / 3 of M, 25), 926.5 (1 / 2 of M, 100), 618.3 (1 / 3 of M, 40).

[0222] Example 4: Synthesis of the SAc - 225 / PSMA - 62 complex (radio - labeled). Ac - 225 (as AcCl3) was prepared in 0.1 M HCl and added to a vial, followed by addition to the vial of reaction buffer and then PSMA - 62 (Formula A - 1 prepared in Example 2 above). The reaction mixture contained 14 MBq / mL of Ac - 225, sodium ascorbate (32 mg / mL), ammonium acetate (15 mg / mL), PSMA - 62 (293 μg / mL), and had a pH of 5.5. The reaction was heated to 90 °C for 15 minutes while shaking (500 rpm), and then quenched with a formulation buffer of DTPA (0.1 mg / mL) and sodium ascorbate (30 mg / mL) in sterile filtered water (final pH was 4.5 - 6.5 and the product RAC was 2 MBq / mL).

[0223] Example 5: Purity analysis of the compound and identification of structural impurities The PSMA-62 compound was prepared as generally described for PSMA-62 in Example 1 of PCT / EP2018 / 084399. The purity of the compound (sample) was evaluated as follows. Sample solvent: DMSO: water (1:10); injection volume: 20 μL. The LCMS method was carried out using an Agilent 1290 / 6460 Triple Quad LC / MS system equipped with a UV detector (monitored at 210 nm) using a Waters Xbridge BEH Phenyl Column, 4.6×150 mm, 3.5 μm (IID 19898). The gradient elution system utilized mobile phase A (0.05% TFA in HPLC grade water, pH 2.02) and mobile phase B (CAN). The gradient started at 85% A and 15% B and was carried out at a flow rate of 1.0 mL / min, increasing to 76% A and 24% B over 20 minutes; then it was increased to 64% A and 36% B over 4 minutes. Finally, the gradient parameters were returned to the initial starting conditions. Data was analyzed using Chemstation software from Agilent. Figure 3 is the HPLC chromatogram obtained, showing impurities at 13.9 minutes and the compound of formula A-3 at 14.4 minutes.

[0224] Example 6: Preparation of Formulations For intravenous injection 225 Preferred batches of the Ac PSMA-62 formulation are prepared using the following formulation components and their amounts shown in Table 3 below. 225 Ac PSMA-62 can be prepared as in Examples 2 and 4 above.

[0225] Table 3: 225 Components of the Ac PSMA-62 injection solution [Table 4] 1 At end of synthesis (EOS) 2225 Contains Ac PSMA-62, PSMA-62, and metal PSMA-62 complexes. Based on a nominal 687.5 μg peptide input to the process. 3A single-dose vial containing a single-dose amount for 1 to 5 administrations (based on the patient's body surface area (BSA) and current dose level (DL)).

[0226] Example 7: Preparation of formulation For injection 225 The Ac PSMA-62 formulation can be prepared as follows. Transfer 225AcCl3 in 0.1N HCl solution (1.4 mL) to a 10 mL reaction vial. 1. Reaction buffer * (0.4 mL) is transferred to a 10 mL reaction vial. 2. Add the PSMA-62 precursor solution (PSMA-62 in 0.04N HCl) (0.55 mL) to the reaction vial. 3. Using a preheated heater / shaker, heat the reaction vial at 90 ± 2 °C for 15 ± 1 minutes while shaking at 500 rpm. 4. Once radiolabeling is complete, filter the entire solution through a 2 × 0.22 μm inline sterilizing filter into a 30 mL DP bulk vial. 5. 10 mL of formulation buffer ** is added to the reaction vial, the contents are mixed thoroughly, and the entire rinse solution is filtered through a 2 × 0.22 μm inline sterilizing filter into a 30 mL drug product (DP) bulk vial. 6. Rinse the sterilizing filter with 6.1 mL of formulation buffer ** so that all the contents are recovered and then transferred to a 30 mL DP bulk vial. 7. To ensure that all the contents are recovered, push air through the sterilizing filter into the 30 mL DP bulk vial. The high-purity 225 formulation containing Ac PSMA-62 thus prepared can be stored at room temperature or a lower temperature until administration to the patient. * Reaction buffer preparation: Prepared by dissolving 1878 mg of sodium ascorbate and 924 mg of ammonium acetate in 10 mL of sterile water for injection (SWFI). **Preparation of formulation buffer: The formulation buffer used in the above protocol is prepared by mixing 596 mg of sodium ascorbate with 2.3 mL of DTPA solution (prepared by dissolving 23 mg of DTPA in 23 mL of sterile water for injection (SWFI)) and 17.7 mL of SWFI.

[0227] An alternative formulation buffer that can be suitably utilized in the above protocol can be prepared as follows: Mix 596 mg of sodium ascorbate with 2.3 mg of DTPA and 20 mL of SWFI.

[0228] Example 8: Treatment protocol A human male patient is selected for treatment after being diagnosed with metastatic castration-resistant prostate cancer (e.g., manifested by disease progression despite surgical or chemical castration) and after other treatments, e.g., by meeting other eligibility criteria including PSMA PET positivity with an existing PSMA imaging agent. 225 Ac PSMA-62 (with a chemical purity of greater than 90% in sterile aqueous solution and appropriately prepared as in Examples 2, 4, and 7 above 225 and determined by Ac PSMA-62) is administered to the patient by intravenous injection. The dosing regimen can include receiving a single dose of Ac PSMA-62 on day 1 of each 6-week cycle for up to 4 cycles. 225

[0229] Example 9: 68 Synthesis of Ga-PSMA 62 68 Ga-PSMA 62 can be appropriately prepared as follows. 68 Gallium (iThema LABS) is provided as 1.2 mL of 0.6 M HCl. 1 mL of this 68 Ga composition is transferred to a reaction vial, and reaction buffer (600 μL) is added to obtain a pH of 4.5. A PSMA-62 (non-chelated) solution (200 μL) is added, and the reaction vial is heated at 90 °C for 10 minutes. After heating, the labeled solution is loaded onto a C18 cartridge to remove the68 Isolate Ga-PSMA 62. Rinse the C18 cartridge with saline (8 mL total, including 2 mL used to rinse the reaction vial). Finally, 68 Ga-PSMA 62 is eluted from the C18 with purification eluent (a 3:1 mixture of formulation buffer and ethanol) and formulation buffer (a 1:1 mixture of 2.5 M NaOAc and 0.9% NaCl) and filtered through a 0.22 μm filter into a final DP vial.

[0230] Example 10: Treatment of patients with mCRPC This example illustrates, inter alia, patient-specific dosimetry for the application of 225Ac-PSMA-62 in patients diagnosed with metastatic castration-resistant prostate cancer. Seven male human patients diagnosed with mCRPC and who showed disease progression after at least one prior standard therapy were administered 225Ac-PSMA-62. 68 Ga-PSMA-62 PET / CT imaging was performed. 225 Eligibility for Ac-PSMA-62 treatment was confirmed. All seven patients received 4MBq of Ac-PSMA-62 during the first cycle. 225 Patients received an intravenous (IV) injection of Ac-PSMA-62. SPECT / CT acquisitions on a Mediso AnyScan gamma camera were performed on one patient, covering two fields of view encompassing the salivary glands and kidneys. Three primary energy windows at 440 keV, 218.20 keV and 92.0 keV with window widths of 20%, 20% and 25%, respectively, were acquired as separate images. Imaging sessions were performed at 3±1 h, 24±3 h and 144±24 h after administration. For the 3 and 24 h images, 120 projections were acquired in a 128×128 matrix with 60 projections per patient at 30 s per projection. The 144 h acquisition involved 45 s per projection. Reconstructions included effective source scatter estimation scatter correction, CT-based attenuation correction, full collimator-detector response and CT-based attenuation correction. 225 We utilized an ordered subset expectation-maximization reconstruction algorithm with crosstalk between all isotopes in the Ac decay chain. 225Since only Ac-PSMA-62 was administered, this is the amount usually administered for radioligand therapy of patients with mCRPC 225 half of the amount of Ac, but the biodistribution of the reconstructed images 68 was compared between Ga-PSMA-62 225 and Ac-PSMA-62.

[0231] Results: 225 The lesion biodistribution observed in the qualitative data of Ac-PSMA-62 68 was comparable to that of the Ga-PSMA-62 results. See the human patient images in Figure 4. Smaller lesions 68 may not be clearly observed with Ga-PSMA-62, but the indication showing target vs non-target was equivalent between the two isotopes (i.e., 68 between Ga-PSMA-62 225 and Ac-PSMA-62).

[0232] Conclusion: This in-human study demonstrates that SPECT / CT imaging of Ac-PSMA-62 at a lower activity of 4 MBq is achievable. Dosimetry analysis using Ac-PSMA-62 at 4 MBq is possible. Further studies may include variable 225 Ac-PSMA-62 activities and administration frequencies. 225 225 225 225

[0233] Example 11: In patients with mCRPC 68 intra-individual comparison of Ga-PSMA-62 and 68 Ga-68 PMSA-11 PET / CT This pilot clinical study of this example presents an intra-individual comparison to evaluate the tracer-specific characteristics of 68 Ga-PSMA-62 vs 68 Ga-PSMA-11. Seven human male patients with metastatic castration-resistant prostate cancer (mCRPC), presented for a potential radioligand, received both Ga-PSMA-62 and 68 Ga-PSMA-62 and 68Underwent Ga-PSMA-11 PET / CT. All patients received similar activities of approximately 185 MBq of 68 Ga-PSMA-62 and 68 Ga-PSMA-11 and were imaged on the same PET / CT scanner using the same reconstruction algorithm. The biodistribution in normal organs and tumor uptake were quantified using SUV max .

[0234] PSMA-affinic lesions in the prostate were identified in all seven patients in local tumors, lymph node metastases, visceral metastases, and bone metastases. For local tumors, lymph node metastases, visceral metastases, and bone metastases, 68 Ga-PSMA-62 and 68 Ga-PSMA-11 SUV max When compared, no statistical significance was observed. Regarding normal organs, 68 Ga-PSMA-62 had lower uptake both visually and semi-quantitatively in the parotid glands. 68 Compared to Ga-PSMA-11 PET / CT images (SUVmax: mean 16.35, median 14.81, range 8.06 - 33.86), 68 Ga-PSMA-62 PET / CT images showed uptake in both parotid glands (SUVmax: mean 9.06, median 8.76, range 4.19 - 21.51). 68 Ga-PSMA-62 PET / CT images also showed 68 A superior tumor:salivary gland ratio compared to Ga-PSMA-11 PET / CT images. (Mean 20.44; median 12.6 and range 2.68 - 68.06) vs. (mean 5.21, median 3.28 and range 1.35 - 12.76).

[0235] This example demonstrated, among other things, that Ga-PSMA-62 68 is comparable to 68 Ga-PSMA-11 in the identification of prostate cancer lesions, especially in patients with mCRPC. In addition, 68 Ga-PSMA-62 has 68 significantly lower salivary gland uptake compared to Ga-PSMA-11.68 This preliminary clinical data with Ga-PSMA-62 is promising and indicates that treatment with 225Ac-PSMA-62 may result in a reduction in the incidence and severity of xerostomia compared to currently available PSMA-targeted radiopharmaceuticals.

[0236] Example 11 Procedure 1:

[0237]

Chem.

[0238]

Chem.

[0239]

Chem.

[0240]

Chem.

[0241]

Chem.

[0242]

Chem.

[0243]

Chem.

[0244] Preparation 1: (9H-Fluoren-9-yl)methyl ((R)-1-(((R)-6-((diphenyl(p-tolyl)methyl)amino)-1-(λ 1-Oxide Anail)-1-oxohexan-2-yl)amino)-3-(naphthalen-2-yl)-1-oxopropan-2-yl)carbamate

[0245]

Chem.

[0246] Fmoc-D-Lys(Mtt)Wang resin (23 g, 0.52 meq / g, 11.96 mmol) was transferred to a 1 L reaction vessel. The resin was washed with DMF (190 mL) for 10 minutes. It was washed twice with a 20% piperidine solution in DMF (280 mL) to remove the Fmoc group. The resin was washed with DMF (6 × 190 mL), and then treated with a solution of Fmoc-D-2-Nal-OH (10.5 g, 24 mmol), ethyl cyano(hydroxyamino)acetate (3.4 g, 23.93 mmol), and diisopropylcarbodiimide (4 mL, 39.68 mmol) in DMF (23 mL). After coupling for about 3.5 hours, the vessel was drained under reduced pressure and washed with DMF (3 × 190 mL) to obtain the title compound.

[0247] Preparation 2: (9H-Fluoren-9-yl)methyl ((R)-3-(4-(tert-butoxy)phenyl)-1-(((R)-1-(((R)-6-((diphenyl(p-tolyl)methyl)amino)-1-(λ 1 -Oxide Anail)-1-oxohexan-2-yl)amino)-3-(naphthalen-2-yl)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)carbamate

[0248]

Chem.

[0249] (9H-Fluoren-9-yl)methyl ((R)-1-(((R)-6-((diphenyl(p-tolyl)methyl)amino)-1-(λ 1The resin containing (11.96 mmol) of (7R,10R,13R,16S)-16-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)-7-(λ-oxidoanilino)-1-oxohexan-2-yl)amino)-3-(naphthalen-2-yl)-1-oxopropan-2-yl)carbamate was washed with DMF (1×190 mL) and then washed twice with 20% piperidine in DMF (280 mL) to remove the Fmoc group. The resin was washed with DMF (6×190 mL) and then treated with Fmoc-D-Tyr(tBu)-OH (11.0 g, 23.93 mmol), ethyl cyano(hydroxyamino)acetate (3.4 g, 23.93 mmol), and diisopropylcarbodiimide (4 mL, 39.68 mmol) in DMF (23 mL). After coupling for about 11.5 h, the vessel was drained under reduced pressure and washed with DMF (3×190 mL) to give the title compound.

[0250] Preparation 3: tert-butyl (4-((7R,10R,13R,16S)-16-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-7-(λ 1 -oxidoanilino)carbonyl)-13-(4-(tert-butoxy)benzyl)-10-(naphthalen-2-ylmethyl)-9,12,15-trioxo-1,1-diphenyl-1-(p-tolyl)-2,8,11,14-tetraazapentadecan-17-yl)phenyl)carbamate

[0251]

Chem.

[0252] (9H-fluoren-9-yl)methyl ((R)-3-(4-(tert-butoxy)phenyl)-1-(((R)-1-(((R)-6-((diphenyl(p-tolyl)methyl)amino)-1-(λ 1-Oxide aneil)-1-oxohexan-2-yl)amino)-3-(naphthalen-2-yl)-1-oxopropan-2-yl)amino)-1-oxopropan-2-yl)carbamate (1196 mmol) was washed with DMF (1×190 mL) and then washed twice with 20% piperidine in DMF to remove the Fmoc group. The resin was washed with DMF (6×190 mL), and then treated with Fmoc-Phe(4-NHBoc)-OH (12.0 g, 23.88 mmol), ethyl cyano(hydroxyamino)acetate (3.4 g, 23.93 mmol), and diisopropylcarbodiimide (4 mL, 39.68 mmol) in DMF (23 mL). After coupling for about 7.5 h, the vessel was drained under reduced pressure and washed with DMF (3×190 mL) to obtain the title compound.

[0253] Preparation 4: Tri-tert-butyl 2,2’,2’’-(10-((7R,10R,13R,16S,21R)-7-(λ 1 -Oxide aneil)carbonyl)-13-(4-(tert-butoxy)benzyl)-16-(4-((tert-butoxycarbonyl)amino)benzyl)-24,24-dimethyl-10-(naphthalen-2-ylmethyl)-9,12,15,18,22-pentaoxo-1,1-diphenyl-1-(p-tolyl)-23-oxa-2,8,11,14,17-pentaazapentacosane-21-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate

[0254]

Chem.

[0255] tert-Butyl (4-((7R,10R,13R,16S)-16-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-7-(λ 1The resin containing [[(11.96 mmol) of phenyl)carbamate of 17-yl)phenyl) of 2,8,11,14-tetraazapentadecane-1-(p-tolyl)-1,1-diphenyl-9,12,15-trioxo-10-(naphthalen-2-ylmethyl)-13-(4-(tert-butoxyl)benzyl)]] was washed with DMF (1×190 mL) and washed twice with 20% piperidine in DMF (280 mL) to remove the Fmoc group. The resin was washed with DMF (6×190 mL) and then treated with (R)-DOTAGA(tBu)4-OH (16.8 g, 23.97 mmol), PyAOP (12.5 g, 23.97 mmol), and DIEA (9 mL, 51.67 mmol) in DMF (69 mL). After coupling for about 3.5 hours, the vessel was drained under reduced pressure and washed with DMF (3×190 mL) to obtain the title compound.

[0256] Preparation 5: Tri-tert-butyl 2,2′,2″-(10-((5R,10S,13R,16R,19R)-19-(λ 1 -oxide aneil)carbonyl)-23-amino-13-(4-(tert-butoxy)benzyl)-10-(4-((tert-butoxycarbonyl)amino)benzyl)-2,2-dimethyl-16-(naphthalen-2-ylmethyl)-4,8,11,14,17-pentaoxo-3-oxa-9,12,15,18-tetraazatricosane-5-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate

[0257]

Chemical formula

[0258] Tri-tert-butyl 2,2′,2″-(10-((7R,10R,13R,16S,21R)-7-(λ 1Resin containing -oxide aneil)carbonyl)-13-(4-(tert-butoxy)benzyl)-16-(4-((tert-butoxycarbonyl)amino)benzyl)-24,24-dimethyl-10-(naphthalen-2-ylmethyl)-9,12,15,18,22-pentaoxo-1,1-diphenyl-1-(p-tolyl)-23-oxa-2,8,11,14,17-pentaazapentacosan-21-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (11.96 mmol) was washed with DCM (2×190 mL) and washed twice with 40% hexafluoroisopropanol in DCM (2×280 mL) to remove the Mtt group. The resin was washed with DCM (4×190 mL) and subsequently washed with 10% DIEA in DMF for neutralization (2×280 mL). The resin was washed with DMF (2×190 mL) to obtain the title compound.

[0259] A sample of this resin was taken out, washed with IPA (3x), and dried. The dried resin was treated with a TFA cleavage cocktail (3 mL; 95% TFA, 5% H2O, 25 mg / mL DTT) and shaken at room temperature for 1.25 hours. The resin was removed by filtration, and the crude peptide was precipitated with cold MTBE (25 mL). The precipitated peptide was centrifuged and the supernatant was decanted. The pellet was washed with cold MTBE (25 mL), dried under nitrogen, capped using a mixture of 25% acetic anhydride and 10% DIEA in DMF to obtain the capped peptide, which was analyzed by MS. The molecular weight of R-DOTAGA(tBu)4-Phe(4-NH2)-D-Tyr-D-2-Nal-D-Lys-NH2 is 1392.72. ES / MS m / z: 1393.7 (M+H).

[0260] Preparation 6: 1-allyl 25-(tert-butyl) (3R,10R,13R,16R,19S,24R)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-10-(λ 1-Oxideanil)carbonyl)-16-(4-(tert-butoxy)benzyl)-19-(4-((tert-butoxycarbonyl)amino)benzyl)-13-(naphthalen-2-ylmethyl)-4,12,15,18,21-pentaoxo-24-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-5,11,14,17,20-pentaazapentacosanedioate

[0261] [Chemical formula]

[0262] Tri-tert-butyl 2,2’,2’’-(10-((5R,10S,13R,16R,19R)-19-(λ 1 -Oxideanil)carbonyl)-23-amino-13-(4-(tert-butoxy)benzyl)-10-(4-((tert-butoxycarbonyl)amino)benzyl)-2,2-dimethyl-16-(naphthalen-2-ylmethyl)-4,8,11,14,17-pentaoxo-3-oxa-9,12,15,18-tetraazatricosane-5-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (11.96 mmol) containing resin was washed with DMF (1×190 mL) and washed twice with 20% piperidine in DMF (280 mL) to remove the Fmoc group. The resin was washed with DMF (6×190 mL) and then treated with Fmoc-D-Asp(OAll)-OH (9.4 g, 23.77 mmol), PyAOP (12.5 g, 23.97 mmol) and DIEA (9 mL, 51.67 mmol) in DMF (23 mL). After coupling for about 14 hours, the vessel was drained under reduced pressure and washed with DMF (3×190 mL) to obtain the title compound.

[0263] Preparation 7: Di-tert-butyl 5-(((5R,10S,13R,16R,19R,26R)-19-((λ 1-Oxide aneil)carbonyl)-13-(4-tert-butoxy)benzyl)-10-(4-((tert-butoxycarbonyl)amino)benzyl-2,2-dimethyl-16-(naphthalen-2-ylmethyl)-4,8,11,14,17,25,28-heptaoxo-5-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-3,29-dioxa-9,12,15,18,24-pentaazadotriacont-31-en-26-yl)carbamoyl)isophthalate

[0264]

Chem.

[0265] 1-Allyl 25-(tert-butyl) (3R,10R,13R,16R,19S,24R)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-10-((λ 1 -Oxide aneil)carbonyl)-16-(4-(tert-butoxy)benzyl)-19-(4-((tert-butoxycarbonyl)amino)benzyl)-13-(naphthalen-2-ylmethyl)-4,12,15,18,21-pentaoxo-24-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-5,11,14,17,20-pentaazapentacosanedioate-containing resin (11.96 mmol) was washed with DMF (1×190 mL) and washed twice with 0.5 M ethyl cyano(hydroxyamino)acetate (19.9 g) in 10% piperidine (28 mL) / DMF (252 mL) (total volume 280 mL) to remove Fmoc. The resin was washed with DMF (6×190 mL) and then treated with TMA(tBu)2 (7.7 g, 23.77 mmol), PyAOP (12.5 g, 23.88 mmol), and DIEA (9 mL, 51.67 mmol) in DMF (23 mL). After coupling for about 17.75 h, the vessel was drained under reduced pressure and washed with DMF (3×190 mL) to obtain the title compound.

[0266] A sample of the resin was taken out before coupling TMA to the peptide, washed with IPA (3x), and dried. The dried resin was treated with a TFA cleavage cocktail (3 mL; 95% TFA, 5% H2O, 25 mg / mL DTT) and shaken and cooled at room temperature for 1.5 h. The resin was removed by filtration, and the crude peptide was precipitated with MTBE (25 mL). The precipitated peptide was centrifuged and the supernatant was decanted. The pellet was washed with cold MTBE (25 mL), dried under nitrogen, and analyzed by MS. The molecular weight of R-DOTAGA(tBu)4-Phe(4-NH2)-D-Tyr-D-2-Nal-D-Lys{D-Asp[OAll]} is 1505.84. ES / MS m / z: 1506 (M+H).

[0267] A sample of the resin was taken out after coupling TMA to the peptide, washed with IPA (3x), and dried. The dried resin was treated with a TFA cleavage cocktail (3 mL; 95% TFA, 5% H2O, 25 mg / mL DTT) and shaken at room temperature for 1.5 h. The resin was removed by filtration, and the crude peptide was precipitated with cold MTBE (25 mL). The precipitated peptide was centrifuged and the supernatant was decanted. The pellet was washed with cold MTBE (25 mL), dried under nitrogen, and analyzed by MS. The molecular weight of (R)-DOTAGA(tBu)4-Phe(4-NH2)-D-Tyr-D-2-Nal-D-Lys{TMA-D-Asp[OAll]} is 1697.97. ES / MS m / z: 1698.5 (M+H).

[0268] Preparation 8: (5R,10S,13R,16R,19R,26R)-19-((λ 1-Oxide aneil)carbonyl)-26-(3,5-bis(tert-butoxycarbonyl)benzamide)-13-(4-(tert-butoxy)benzyl)-10-(4-((tert-butoxycarbonyl)amino)benzyl)-2,2-dimethyl-16-(naphthalen-2-ylmethyl)-4,8,11,14,17,25-hexaoxo-5-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-3-oxa-9,12,15,18,24-pentaazaoctacosan-28-oic acid

[0269]

Chem.

[0270] Di-tert-butyl 5-(((5R,10S,13R,16R,19R,26R)-19-((λ 1 -Oxide aneil)carbonyl)-13-(4-(tert-butoxyl)benzyl)-10-(4-((tert-butoxycarbonyl)amino)benzyl)-2,2-dimethyl-16-(naphthalen-2-ylmethyl)-4,8,11,14,17,25,28-heptaoxo-5-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-329-dioxa-9,12,15,18,24-pentaazadotriacont-31-en-26-yl)carbamoyl)isophthalate-containing resin (11.96 mmol) was washed with DCM (2 × 190 mL). Removal of the allyl group was initiated by treating the resin with tetrakis(triphenylphosphine)palladium(0) (1.4 g, 1.21 mmol) in DCM (100 mL). After about 6 minutes, the reaction was treated with 1,3-dimethylbarbituric acid (18.7 g, 119.76 mmol) in DMF (100 mL), the reaction vessel was wrapped with Al foil, and the reaction was allowed to proceed for about 16.75 hours. The foil was removed, and the resin was washed with DCM (4 × 190 mL). The pH of the resin was neutralized with 5% DIEA in DMF (280 mL), and the resin was washed with DMF (2 × 190 mL) to afford the title compound.

[0271] A sample of the resin after allyl group deprotection was taken out, washed with IPA (3x), and dried. The dried resin was treated with a TFA cleavage cocktail (3 mL; 95% TFA, 5% H2O, 25 mg / mL DTT) and shaken and cooled at room temperature for 1.5 hours. The resin was removed by filtration, and the crude peptide was precipitated with cold MTBE (25 mL). The precipitated peptide was centrifuged, and the supernatant was decanted. The pellet was washed with cold MTBE (25 mL), dried under nitrogen, and analyzed by MS. The exact mass of R-DOTAGA(tBu)4-Phe(4-NH2)-D-Tyr-D-2-Nal-D-Lys{D-Asp[OH]} is 1657.82. ES / MS m / z: 1658.7 (M+H).

[0272] Preparation 9: Di-tert-butyl 5-(((5R,12R,19R,22R,25R,28S,33R)-19-((λ 1 -oxide anil)carbonyl)-25-(4-tert-butoxy)benzyl-5-(tert-butoxycarbonyl)-28-(4-((tert-butoxycarbonyl)amino)benzyl)-1-(9H-fluoren-9-yl)-36,36-dimethyl-22-(naphthalen-2-ylmethyl)-3,10,13,21,24,27,30,34-octaoxo-33-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-2,35-dioxa-4,9,14,20,23,26,29-heptaazapentatriacontan-12-yl)carbamoyl)isophthalate

[0273]

Chemical formula

[0274] (5R,10S,13R,16R,19R,26R)-19-(λ 1-Oxide aneil)carbonyl)-26-(3,5-bis(tert-butoxycarbonyl)benzamide)-13-(4-(tert-butoxy)benzyl)-10-(4-((tert-butoxycarbonyl)amino)benzyl)-2,2-dimethyl-16-(naphthalen-2-ylmethyl)-4,8,11,14,17,25-hexaoxo-5-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-3-oxa-9,12,15,18,24-pentaazaoctacosan-28-oic acid (11.96 mmol) containing resin was washed with DMF (1×190 mL). The resin was treated with Fmoc-D-Orn-OtBu hydrochloride (10.7 g, 23.94 mmol), PyAOP (12.5 g, 23.88 mmol) and DIEA (13 mL, 74.63 mmol) in DMF (69 mL). After coupling for about 22.75 h, the vessel was drained under reduced pressure and washed with DMF (3×190 mL) to give the title compound.

[0275] A sample of the residue after coupling D-Orn to the peptide was removed, washed with IPA (3x) and dried. The dried resin was treated with TFA cleavage cocktail (3 mL; 95% TFA, 5% H2O, 25 mg / mL DTT) and shaken at room temperature for 1.5 h. The resin was removed by filtration and the crude peptide was precipitated with cold MTBE (25 mL). The precipitated peptide was centrifuged and the supernatant was decanted. The pellet was washed with cold MTBE (25 mL), dried under nitrogen and analyzed by MS. The molecular weight of R-DOTAGA(tBu)4-Phe(4-NH2)-D-Tyr-D-2-Nal-D-Lys{TMA-D-Asp[dD-Orn-OH]} is 1773. ES / MS m / z: 887.2 ((M+2H) / 2).

[0276] Preparation 10: Penta-tert-butyl (3S,7S,12R,19R,26R,29R,32R,35S,40R)-26-(λ 1Oxide aneil)carbonyl)-19-(3,5-bis(tert-butoxycarbonyl)benzamide)-32-(4-(tert-butoxy)benzyl)-35-(4-((tert-butoxycarbonyl)amino)benzyl)-29-(naphthalen-2-ylmethyl)-5,10,17,20,28,31,34,37-octaoxo-40-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-4,6,11,16,21,27,30,33,36-nonaazatetracontane-1,3,7,12,40-pentacarboxylate [(R)-DOTAGA(tBu)-Phe(4-NHBoc)-D-Tyr(tBu)-D-Nal-D-Lys{TMA-D-Asp[δD-Orn-OtBu(γGlu(tBu)-CO-Glu(tBu)-OtBu]}-Wang resin]

[0277] [Chemical formula]

[0278] Di-tert-butyl 5-(((5R,12R,19R,22R,25R,28S,33R)-19-(λ 1-(Oxide aneil)carbonyl)-25-(4-(tert-butoxy)benzyl)-5-(tert-butoxycarbonyl)-28-(4-((tert-butoxycarbonyl)amino)benzyl)-1-(9H-fluoren-9-yl)-36,36-dimethyl-22-(naphthalen-2-ylmethyl)-3,10,13,21,24,27,30,34-octaoxo-33-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-2,35-dioxa-4,9,14,20,23,26,29-heptaazaoctatriacontan-12-yl)carbamoyl)isophthalate (11.96 mmol) containing resin was washed with DMF (1 × 190 mL), and washed twice with 20% piperidine in DMF (280 mL) to remove the Fmoc group. The resin was washed with DMF (6 × 190 mL), and treated with (S)-5-(tert-butoxy)-4-(3-((S)-1,5-di-tert-butoxy-1,5-dioxopentan-2-yl)ureido)-5-oxopentanoic acid (11.7 g, 23.95 mmol, abbreviated as (tBuO)EuE(OtBu)2 elsewhere), PyAOP (12.5 g, 23.88 mmol), and DIEA (9 mL, 51.67 mmol) in DMF (69 mL). After coupling for about 16 hours, the container was drained under reduced pressure, the resin was washed with DMF (4 × 190 mL), and washed twice with a DMF (190 mL) solution of sodium diethylthiocarbamate (1.04 g) at 5 g / L to obtain the title compound after drying.

[0279] Example 1: (3S,7S,12R,19R,26R,29R,32R,35S,40R)-35-(4-aminobenzyl)-19-(3,5-dicarboxybenzamide)-32-(4-hydroxybenzyl)-29-(naphthalen-2-ylmethyl)-5,10,17,20,28,31,34,37-octaoxo-40-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-4,6,11,16,21,27,30,33,36-nonaazatetracontane-1,3,7,12,26,40-hexacarboxylic acid

[0280]

Chem.

[0281] Penta-tert-butyl (3S,7S,12R,19R,26R,29R,32R,35S,40R)-26-((λ 1 -oxide aneil)carbonyl)-19-(3,5-bis(tert-butoxycarbonyl)benzamide)-32-(4-(tert-butoxy)benzyl)-35-(4-((tert-butoxycarbonyl)amino)benzyl)-29-(naphthalen-2-ylmethyl)-5,10,17,20,28,31,34,37-octaoxo-40-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)-4,6,11,16,21,27,30,33,36-nonaazatetracontane-1,3,7,12,40-pentacarboxylate (23.66 g) containing resin was carefully added to a peptide cleavage solution (prepared by mixing TFA (230 mL), water (2.5%; 6 mL), triisopropylsilane (2.5%; 6 mL), and dithiothreitol (0.025 g / mL; 6 g)) that had been pre-cooled to -10 °C to 0 °C and stirred for 10 minutes. After the addition of the resin, the cooling bath was removed and the mixture was stirred for about 17.75 hours. The reaction mixture was filtered through an HDPE frit funnel and the resin was washed with TFA (2 × 24 mL). The filtrate was concentrated to 120 mL under reduced pressure and transferred to a 2 L Erlenmeyer flask. The concentration flask was washed with 10 mL of TFA and the washings were added to the Erlenmeyer flask. The stirred peptide solution was cooled to 2 °C in an ice bath and slowly treated with cooled MTBE (960 mL; -18 °C). The ice bath was removed, the reaction mixture was stirred for 35 minutes and then allowed to stand for 60 minutes. Approximately 450 mL of the solvent was decanted and then the deprotected peptide was removed by filtration through a glass frit funnel (600 mL) and the product was washed with MTBE (2 × 192 mL). The crude material was dried under vacuum to give the crude peptide (11.86 g, 83%). ES / MS (m / z): 926 ([M+2H] / 2).

[0282] Purification of crude PSMA-62: The crude PSMA-62 (Formula A-1) from the cleavage and overall deprotection steps was purified by a two-step preparative reversed-phase HPLC procedure on C18 derivatized silica. The purification was carried out on a 5 cm NovaSep column packed with 300 g of Daisogel SP-120-10-ODS-RPS C18 media. The elution from the column was monitored at 254 nm UV, and the resulting fractions were analyzed by an in-process RP-HPLC method.

[0283] Step 1: Purification by preparative reversed-phase chromatography (RPC1) In the first stage, the crude peptide was dissolved in an aqueous solution of 0.84 M ammonium bicarbonate (NH4HCO3) and filtered through a new or dedicated high-density polyethylene (HDPE) frit funnel. The column was equilibrated with 3 column volumes (CV) of 2% ACN in 0.1 M NH4HCO3 (6 L). The product was eluted over 120 minutes using a gradient of 2% - 20% ACN in 0.1 M NH4HCO3. To avoid peptide degradation, all fractions were immediately acidified with neat TFA, and the fractions containing the product with a purity of 94% or higher were pooled and subjected to a second reversed-phase purification.

[0284] Step 2: Purification by preparative reversed-phase chromatography (RPC2) The fractions pooled from the first reversed-phase purification were diluted 1:2 with water and loaded onto the same NovaSep column, but equilibrated with 10% ACN in 0.1% aqueous TFA. The peptide was eluted over 120 minutes using a gradient of 10% ACN - 30% ACN in 0.1% aqueous TFA. The fractions containing the product with a purity of 95% or higher were pooled, filtered, shell frozen, and lyophilized to produce the product in the form of a dry powder as the TFA salt.

[0285] Final lyophilization The lyophilized lot from the second reverse-phase purification was reconstituted in 1% TFA / 10% ACN / 89% water. The solubilized peptide was then homogenized, filtered through a 0.2 μm polyvinylidene (PVDF) filter, dispensed into lyophilization flasks (less than 300 mL per flask), shell frozen using solid CO2 / isopropanol refrigerant, and lyophilized at less than -60 °C and less than 500 mTorr for 48 to 168 hours.

[0286] Embodiment Embodiment 1. The following formula A-2:

[0287]

Chemical formula

[0288] Embodiment 2. The following formula A-2-R:

[0289]

Chemical formula

[0290] Embodiment 3. The following formula A-2-S:

[0291]

Chemical formula

[0292] Embodiment 4. An effective amount of the following formula A-3:

[0293]

Chemical formula

[0294] Embodiment 5. An effective amount of the following formula A-3-R:

[0295]

Chemical formula

[0296] Embodiment 6. An effective amount of the following formula A-3-S:

[0297]

Chemical formula

[0298] Embodiment 7. The pharmaceutical composition according to any one of Embodiments 1 to 6, wherein the compound has a chemical purity of 96% or more.

[0299] Embodiment 8. The pharmaceutical composition according to any one of Embodiments 1 to 7, wherein the compound has a chemical purity of 98% or more.

[0300] Embodiment 9. The pharmaceutical composition according to any one of Embodiments 1 to 8, wherein the compound has one or more structural impurities of less than 10%.

[0301] Embodiment 10. The pharmaceutical composition according to any one of Embodiments 1 to 9, wherein the compound has one or more structural impurities of less than 5%.

[0302] Embodiment 11. The pharmaceutical composition according to any one of Embodiments 1 to 10, wherein the compound has less than 10% of impurities containing at least one of the following formulas B-1, B-2, and B-3:

[0303]

Chemical formula

[0304] Embodiment 12. The pharmaceutical composition according to any one of Embodiments 1 to 11, wherein the compound has less than 10% of impurities containing at least one compound of the following formulas B-1-R, B-2-R, and B-3-R:

[0305] [Chemical formula]

[0306] Embodiment 13. The pharmaceutical composition according to any one of Embodiments 1 to 11, wherein the compound has less than 10% of impurities containing at least one compound of the following formulas B-1-S, B-2-S, and B-3-S:

[0307] [Chemical formula]

[0308] Embodiment 14. The pharmaceutical composition according to any one of Embodiments 1 to 13, wherein the compound has less than 10% of one or more impurities having a retention time within 2 minutes of the peak of the compound as evaluated by high performance liquid chromatography.

[0309] Embodiment 15. The pharmaceutical composition according to any one of Embodiments 1 to 13, wherein the compound has less than 5% of one or more impurities having a retention time within 2 minutes of the peak of the compound as evaluated by high performance liquid chromatography.

[0310] Embodiment 16. The pharmaceutical composition according to any one of Embodiments 1, 2, or 7 to 15, wherein Z is 177 Lu, 225 Ac, 211 At, 67 Cu, 161 Tb, 67 Ga, 203 Pb, 223 Ra, and / or 212 Pb.

[0311] Embodiment 17. Z is​​225 The pharmaceutical composition according to embodiment 1, or any one of embodiments 7 to 15, which is Ac.

[0312] In embodiment 18, Z is 177 The pharmaceutical composition according to embodiment 1, 2, or any one of embodiments 7 to 15, which is Lu.

[0313] Embodiment 19. The pharmaceutical composition according to any one of embodiments 1 to 18, wherein the composition comprises one or more stabilizer compounds.

[0314] Embodiment 20. The pharmaceutical composition according to any one of embodiments 1 to 19, wherein the composition comprises one or more ascorbate compounds.

[0315] Embodiment 21. The pharmaceutical composition according to embodiment 20, wherein one or more ascorbate compounds are present in an amount of 10 to 40 mg / mL of the composition.

[0316] Embodiment 22. The pharmaceutical composition according to any one of embodiments 1 to 21, wherein the composition is formulated as an aqueous composition.

[0317] Embodiment 23. A method for treating a subject suffering from metastatic castration-resistant prostate cancer (mCRPC), comprising administering to the subject an effective amount of the following formula A-2:

[0318]

Chemical formula

[0319] Embodiment 24. A method for treating a subject suffering from biochemical recurrence (BCR) prostate cancer, comprising administering to the subject an effective amount of the following formula A-2:

[0320]

Chemical formula

[0321] Embodiment 25. A method for treating a subject suffering from oligometastatic hormone-sensitive prostate cancer (OmHSPC), comprising administering to the subject an effective amount of the following formula A-2:

[0322]

Chemical formula

[0323] Embodiment 26. An effective amount of the following formula A-2-R:

[0324]

Chemical formula

[0325] Embodiment 27. An effective amount of the following formula A-2-R:

[0326]

Chemical formula

[0327] Embodiment 28. An effective amount of the following formula A-3:

[0328]

Chemical formula

[0329] Embodiment 29. An effective amount of the following formula A-3-R:

[0330] [Chemical] The method according to any one of Embodiments 23 to 25, wherein a compound of is administered to a subject.

[0331] Embodiment 30. An effective amount of the following formula A-3-S:

[0332] [Chemical] The method according to any one of Embodiments 23 to 25, wherein a compound of is administered to a subject.

[0333] Embodiment 31. The method according to any one of Embodiments 23 to 30, wherein the administered compound has a chemical purity of more than 90%.

[0334] Embodiment 32. The method according to any one of Embodiments 23 to 31, wherein the administered compound has a chemical purity of more than 95%.

[0335] Embodiment 33. The method according to any one of Embodiments 23 to 32, wherein the administered compound has a chemical purity of more than 98%.

[0336] Embodiment 34. The method according to any one of Embodiments 23 to 33, wherein the compound has one or more structural impurities of less than 10%.

[0337] Embodiment 35. The method according to any one of Embodiments 22 to 34, wherein the compound has one or more structural impurities of less than 5%.

[0338] Embodiment 36. The compound has less than 10% of the following formulas B-1, B-2, and B-3:

[0339] [Chemical] The method according to any one of Embodiments 22 to 35, wherein the compound has impurities containing at least one compound of .

[0340] Embodiment 37. The method according to any one of Embodiments 23 to 36, wherein the compound has impurities containing at least one compound of less than 10% of the following formulas B-1-R, B-2-R, and B-3-R:

[0341] [Chemical formula]

[0342] Embodiment 38. The method according to any one of Embodiments 23 to 37, wherein an aqueous formulation containing the compound is administered to a subject.

[0343] Embodiment 39. The method according to Embodiment 38, wherein the formulation contains one or more stabilizer compounds.

[0344] Embodiment 40. The method according to Embodiment 38 or 39, wherein the formulation contains one or more ascorbic acid compounds.

[0345] Embodiment 41. A method for treating a subject suffering from cancer, comprising: administering to the subject an effective amount of the pharmaceutical composition according to any one of Embodiments 1 to 22.

[0346] Embodiment 42. The method according to Embodiment 41, wherein the subject is suffering from prostate cancer.

[0347] Embodiment 43. A method for treating a subject suffering from metastatic castration-resistant prostate cancer (mCRPC), comprising: administering to the subject an effective amount of the pharmaceutical composition according to any one of Embodiments 1 to 22.

[0348] Embodiment 44. A method for treating a subject suffering from biochemical recurrence (BCR) prostate cancer, comprising: administering to the subject an effective amount of the pharmaceutical composition according to any one of Embodiments 1 to 22.

[0349] ​ Embodiment 45. A method for treating a subject suffering from oligometastatic hormone-sensitive prostate cancer, comprising: administering to the subject an effective amount of the pharmaceutical composition according to any one of Embodiments 1 to 22.

[0350] Embodiment 46. The method according to any one of Embodiments 23 to 45, wherein the subject is evaluated for and identified as having high or elevated prostate-specific membrane antigen (PSMA) expression compared to a healthy subject.

[0351] Embodiment 47. The method according to Embodiment 46, wherein the compound or composition is administered to the identified subject.

[0352] Embodiment 48. The method according to any one of Embodiments 23 to 47, wherein the subject is evaluated by imaging prior to administration of the compound or composition.

[0353] Embodiment 49. The method according to any one of Embodiments 23 to 48, wherein the subject suffers from metastatic castration-resistant prostate cancer (mCRPC) having prostate-specific membrane antigen (PSMA) highly-accumulating lesions.

[0354] Embodiment 50. The method according to any one of Embodiments 23 to 49, wherein the subject suffers from biochemical recurrence (BCR) prostate cancer having prostate-specific membrane antigen (PSMA) highly-accumulating lesions.

[0355] Embodiment 51. The method according to any one of Embodiments 23 to 50, wherein the subject suffers from oligometastatic hormone-sensitive prostate cancer (OmHSPC) having prostate-specific membrane antigen (PSMA) highly-accumulating lesions.

[0356] Embodiment 52. The method according to any one of Embodiments 23 to 51, wherein the subject has been treated with one or more other cancer therapies prior to administration of the compound or composition.

[0357] Embodiment 53. The method according to any one of Embodiments 23 to 52, wherein the subject has been treated with one or more androgen receptor inhibitors before administration of the compound or composition.

[0358] Embodiment 54. A method for treating a subject, comprising: administering to the patient a compound of Formula A-2:

[0359]

Chemical formula

[0360] Embodiment 55. The method according to Embodiment 52, wherein the compound is 68 Ga-PSMA-62.

[0361] Embodiment 56. The method according to Embodiment 54 or 55, wherein the compound is detected in vitro.

[0362] Embodiment 57. The method according to any one of Embodiments 54 to 56, wherein the compound is detected by PET.

[0363] Embodiment 58. 225 The method according to any one of Embodiments 54 to 57, wherein Ac-PSMA-62 is administered to the subject after detecting the compound.

[0364] Embodiment 59. A compound of Formula A-1

[0365]

Chemical formula

[0366]

Chemical formula

[0367] Embodiment 60. The composition according to Embodiment 59, wherein the compound of formula A-1 is at least 95% by weight of the composition.

[0368] Embodiment 61. The composition according to Embodiment 59, wherein the compound of formula A-1 is at least 96% by weight of the composition.

[0369] Embodiment 62. The following formula A-2-R:

[0370]

Chemical formula

[0371]

Chemical formula

[0372] Embodiment 63. The composition according to Embodiment 62, wherein the compound of formula A-2-R is at least 95% by weight of the composition.

[0373] Embodiment 64. Z is 47 Sc, 51 Cr, 57 Co, 58 Co, 59 Fe, 67 Cu, 67 Ga, 68 Ga, 71 Ge, 72 As, 72 Se, 77 As, 77 Br, 81 Rb, 88 Y, 90 Y, 97 Ru, 99m Tc, 100 Pd, 101m Rh, 103 Pb105 Rh, 109 Pd, 111 Ag, 111 In, 113 In, 119 Sb, 121 Sn, 127 Cs, 128 Ba, 129 Cs, 131 Cs, 131 I, 139 La, 140 La, 142 Pr, 143 Pr, 149 Pm, 151 Eu, 153 Eu, 153 Sm, 159 Gr, 161 Tb, 165 Dy, 166 Ho, 169 Eu, 175 Yb, 177 Lu, 186 Re, 188 Re, 189 Re, 191 Os, 193 Pt, 194 Ir, 197 Hg, 198 Au, 199 Ag, 199 Au, 201 Tl, 203 Pb, 211 At, 212 Bi, 212 Pb, 213 Bi, 225 Ac, and 227 the composition according to embodiment 63, selected from the group consisting of Th.

[0374] Embodiment 65. Z is 177 Lu, 225 Ac, 211 At, 67 Cu, 161 Tb, 67 Ga, 68 Ga, 203 Pb, 223 Ra, and 212 the composition according to embodiment 63, selected from the group consisting of Pb.

[0375] Embodiment 66. Z is 225 Ac, the composition according to Embodiment 62 or 63.

[0376] Embodiment 67. Z1 is absent, the composition according to any one of Embodiments 62, 63, 64, 65, or 66.

[0377] Embodiment 68. Z1 is the same as Z, the composition according to any one of Embodiments 62, 63, 64, 65, or 66.

[0378] Embodiment 69. A purified compound of formula A-1: wherein the following formula:

[0379]

Chemical formula

[0380]

Chemical formula

[0381] Embodiment 70. A purified compound of the following formula:

[0382]

Chemical formula

[0383]

Chemical formula

[0384] Embodiment 71. A purified compound of the following formula:

[0385]

Chemical formula

[0386] Embodiment 72. Purified compound of formula A-1:

[0387]

Chemical formula

[0388]

Chemical formula

[0389]

Chemical formula

[0390] Embodiment 73. A compound of formula A-1:

[0391]

Chemical formula

[0392]

Chemical formula

[0393]

Chemical formula

[0394] ​Compound according to Embodiment 73, wherein the acid comprises acetic acid, trifluoroacetic acid, p-toluenesulfonic acid, HCl, H2SO4, methanesulfonic acid, or trifluoromethanesulfonic acid.

[0395] Compound according to any one of Embodiments 73 or 74, wherein the hydrolysis solvent comprises water.

[0396] Compound according to Embodiment 74, wherein the hydrolysis solvent comprises water and triisopropylsilane.

[0397] Compound according to Embodiment 74, wherein the hydrolysis solvent comprises water and dithiothreitol. In a further embodiment, the hydrolysis solvent comprises water, dithiothreitol, and triisopropylsilane.

[0398] Embodiment 78. A method for preparing a compound of formula X:

[0399]

Chemical formula

[0400]

Chemical formula

[0401]

Chemical formula

[0402]

Chemical formula

[0403] Embodiment 79. A compound of formula X is treated with an acid in a hydrolysis solvent to form a compound of formula A-1:

[0404]

Chemical formula

[0405] Embodiment 80. The method according to Embodiment 78 or 79, wherein the solid support comprises polystyrene cross-linked with divinylbenzene.

[0406] Embodiment 81. The coupling of the compounds of formula Y and formula Z comprises combining the compounds of formula Y and formula Z with a coupling agent in a coupling solvent. The method according to any one of Embodiments 78 to 80.

[0407] Embodiment 82. The method according to Embodiment 81, wherein the coupling agent contains phosphorus.

[0408] Embodiment 83. The coupling agent is (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), benzotriazolyl-oxy-tris[pyrrolidino]-phosphonium hexafluorophosphate (PyBOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP), hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT). The method according to Embodiment 81 or 82, which is selected from the group consisting of.

[0409] Embodiment 84. The method according to embodiment 81, wherein the coupling agent comprises 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 1-hydroxy-7-azabenzotriazole (HOAT), or ethyl cyano(hydroxyimino)acetate (oxyma), diisopropylcarbodiimide (DIC), or dicyclohexylcarbodiimide (DCI).

[0410] Embodiment 85. The method according to any one of embodiments 81 to 84, wherein the coupling solvent comprises dichloromethane or dimethylformamide.

[0411] Embodiment 86. The method according to any one of embodiments 78 to 85, wherein formula Y and formula Z further comprise a base.

[0412] Embodiment 87. The method according to embodiment 86, wherein the base comprises triethylamine or diisopropylethylamine.

[0413] Embodiment 88. The method according to any one of embodiments 79 to 87, wherein the acid comprises at least one of trifluoroacetic acid, hydrochloric acid, or para-toluenesulfonic acid, and the solvent comprises water.

[0414] Embodiment 89. The method according to embodiment 88, wherein the hydrolysis solvent further comprises at least one of triisopropylsilane or dithiothreitol.

[0415] Embodiment 90. The method according to any one of embodiments 79 to 89, wherein the acid comprises trifluoroacetic acid and the hydrolysis solvent comprises water.

[0416] Embodiment 91. The method according to any one of embodiments 79 to 89, wherein the acid comprises trifluoroacetic acid, the hydrolysis solvent comprises water, triisopropylsilane, and dithiothreitol, the coupling agent comprises PyAOP, the coupling solvent comprises dimethylformamide, and the coupling further comprises a base which is N,N-diisopropylethylamine.

[0417] Embodiment 92. The method according to any one of Embodiments 79 to 91, wherein the compound of formula A-2-R or a pharmaceutically acceptable salt thereof is prepared by treating a compound of formula A-1 with a radioactive cation to form a compound of formula A-2-R:

[0418]

Chemical formula

[0419] Embodiment 93. Z is 47 Sc, 51 Cr, 57 Co, 58 Co, 59 Fe, 67 Cu, 67 Ga, 68 Ga, 71 Ge, 72 As, 72 Se, 77 As, 77 Br, 81 Rb, 88 Y, 90 Y, 97 Ru, 99m Tc, 100 Pd, 101m Rh, 103 Pb, 105 Rh, 109 Pd, 111 Ag, 111 In, 113 In, 119 Sb, 121 Sn, 127 Cs, 128 Ba, 129 Cs, 131 Cs, 131 I, 139 La, 140 La, 142 Pr, 143 Pr, 149 Pm, 151 Eu, 153 Eu, 153 Sm, 159 Gr, 161 Tb, 165 Dy, 166 Ho, 169Eu, 175 Yb, 177 Lu, 186 Re, 188 Re, 189 Re, 191 Os, 193 Pt, 194 Ir, 197 Hg, 198 Au, 199 Ag, 199 Au, 201 Tl, 203 Pb, 211 At, 212 Bi, 212 Pb, 213 Bi, 225 Ac, and 227 The method according to embodiment 92, selected from the group consisting of Th.

[0420] Embodiment 94. Z is, 177 Lu, 225 Ac, 211 At, 67 Cu, 161 Tb, 67 Ga, 68 Ga, 203 Pb, 223 Ra, and 212 The method according to embodiment 92, selected from the group consisting of Pb.

[0421] Embodiment 95. Z is, 225 Ac, the method according to any one of embodiments 92 to 94.

[0422] Embodiment 96. Treating formula A-1 with a radioactive cation is carried out in an aqueous HCl solution 225 A solution containing AcCl3, an aqueous buffer containing sodium ascorbate and ammonium acetate, and formula A-1, the method according to embodiment 95.

[0423] Embodiment 97. Formula Y:

[0424]

Chemical formula

[0425] [Chemical formula] The method according to any one of Embodiments 78 to 91, which is prepared by deprotecting a compound of (wherein PG is an amine protecting group).

[0426] Embodiment 98. The method according to Embodiment 97, wherein PG is selected from fluorenylmethoxycarbonyl (Fmoc), benzyl (Bn), and benzyloxycarbonyl (Cbz).

[0427] Embodiment 99. The method according to Embodiment 97, wherein PG is fluorenylmethoxycarbonyl (Fmoc), and the deprotection of the formula Y2 comprises treating Y2 with a base in an amine deprotection solvent.

[0428] Embodiment 100. The method according to Embodiment 99, wherein the base is selected from the group consisting of cyclohexylamine, ethanolamine, piperidine, piperazine, triethylamine, and N,N - diisopropylethylamine, and the amine deprotection solvent comprises at least one of dimethylformamide (DMF), dichloromethane, toluene, or N - methyl - 2 - pyrrolidone (NMP).

[0429] Embodiment 101. The method according to Embodiment 99, wherein the base comprises piperidine and the amine deprotection solvent comprises dimethylformamide.

[0430] Embodiment 102. The compound of the formula Y2 is of the formula Y3:

[0431] [Chemical formula] The method according to any one of Embodiments 97 to 101, which is prepared by coupling a compound of (wherein PG is an amine protecting group) with PG - D - Orn - OtBu. One preferred PG group is Fmoc.

[0432] Embodiment 103. The method according to Embodiment 102, wherein the coupling occurs in a coupling solvent in the presence of a coupling agent.

[0433] Embodiment 104. The method according to Embodiment 103, wherein the coupling agent contains phosphorus.

[0434] Embodiment 105. The method according to Embodiment 104, wherein the coupling agent is selected from the group consisting of (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), benzotriazolyl-oxy-tris[pyrrolidino]-phosphonium hexafluorophosphate (PyBOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP), hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT).

[0435] Embodiment 106. The method according to Embodiment 103, wherein the coupling agent contains 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 1-hydroxy-7-azabenzotriazole (HOAT), or ethyl cyano(hydroxyimino)acetate (oxyma), diisopropylcarbodiimide (DIC), or dicyclohexylcarbodiimide (DCI).

[0436] Embodiment 107. The method according to any one of Embodiments 103 to 106, wherein the coupling solvent contains dichloromethane or dimethylformamide.

[0437] Embodiment 108. The method according to any one of Embodiments 103 to 107, wherein the coupling of the compound of Formula Y3 and PG-D-Orn-OtBu further comprises a base. In a preferred embodiment, PG is Fmoc. When PG is Fmoc, PG-D-Orn-OtBu is tert-butyl (R)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-aminopentanoate.

[0438] Embodiment 109. The method according to Embodiment 108, wherein the base comprises triethylamine or diisopropylethylamine.

[0439] Embodiment 110. The compound of Formula Y3 is of Formula Y4:

[0440] [Chemical formula] (wherein PG2 is a protecting group), and the deprotection is carried out in the presence of a carboxyl deprotecting agent and a carboxyl deprotecting solvent. The method according to any one of Embodiments 102 to 109.

[0441] Embodiment 111. The method according to Embodiment 110, wherein PG2 is selected from the group consisting of allyl, benzyl, and benzhydryl.

[0442] Embodiment 112. The method according to Embodiment 110 or 111, wherein the carboxyl deprotecting solvent comprises at least one of dichloromethane, dimethylformamide, tetrahydrofuran, or ethanol.

[0443] Embodiment 113. The method according to any one of Embodiments 110 to 112, wherein the carboxyl deprotecting solvent comprises dichloromethane and dimethylformamide.

[0444] Embodiment 114. The method according to any one of Embodiments 110 to 113, wherein PG2 is allyl and the carboxyl deprotecting agent comprises Pd.

[0445] Embodiment 115. The method according to any one of Embodiments 110 to 114, wherein PG2 is allyl and the carboxyl deprotecting agent contains Pd(PPh3)4.

[0446] Embodiment 116. The method according to any one of Embodiments 110 to 113, wherein PG2 is allyl, the carboxyl deprotecting agent contains Pd(PPh3)4, and the deprotection further contains a nucleophile.

[0447] Embodiment 117. The method according to Embodiment 116, wherein the nucleophile is selected from the group consisting of 1,3-dimethylbarbituric acid and triphenylphosphine.

[0448] Embodiment 118. The compound of formula Y4 is of formula Y5:

[0449]

Chemical formula

[0450] Embodiment 119. The method according to Embodiment 118, wherein the coupling agent is selected from the group consisting of (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), benzotriazolyl-oxy-tris[pyrrolidino]-phosphonium hexafluorophosphate (PyBOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP), hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT).

[0451] Embodiment 120. The method according to embodiment 118, wherein the coupling agent comprises 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 1-hydroxy-7-azabenzotriazole (HOAT), or ethyl cyano(hydroxyimino)acetate (oxyma), diisopropylcarbodiimide (DIC), or dicyclohexylcarbodiimide (DCI).

[0452] Embodiment 121. The method according to any one of embodiments 118 to 120, wherein the coupling solvent comprises dichloromethane or dimethylformamide.

[0453] Embodiment 122. The method according to any one of embodiments 118 to 121, wherein the coupling of the compound of formula Y5 and 3,5-bis(tert-butoxycarbonyl)benzoic acid further comprises a base.

[0454] Embodiment 123. The method according to embodiment 122, wherein the base comprises triethylamine or diisopropylethylamine.

[0455] Embodiment 124. The compound of formula Y5 is of formula Y6:

[0456]

Chemical formula

[0457] Embodiment 125. The method according to embodiment 124, wherein PG3 is selected from fluorenylmethoxycarbonyl (Fmoc), benzyl, and benzyl chloroformate.

[0458] Embodiment 126. The method according to embodiment 124, wherein PG is fluorenylmethoxycarbonyl (Fmoc), and the deprotection of formula Y6 comprises treating Y6 with an amine deprotecting agent base in an amine deprotecting solvent.

[0459] Embodiment 127. The method according to Embodiment 126, wherein the amine deprotecting base is selected from the group consisting of cyclohexylamine, ethanolamine, piperidine, piperazine, triethylamine, and N,N-diisopropylethylamine, and the amine deprotecting solvent contains at least one of dimethylformamide, dichloromethane, toluene, or N-methyl-2-pyrrolidone (NMP).

[0460] Embodiment 128. The method according to Embodiment 126, wherein the amine deprotecting base contains piperidine and the amine deprotecting solvent contains dimethylformamide.

[0461] Embodiment 129. The compound of formula Y6 is prepared by coupling with PG3-D-Asp(O-PG2)-OH (wherein PG2 and PG3 are different) in a coupling solvent in the presence of a coupling agent, the method according to any one of Embodiments 124 to 128.

[0462] [Chemical formula] The method according to any one of Embodiments 124 to 128, which is prepared by coupling the compound of with PG3-D-Asp(O-PG2)-OH (wherein PG2 and PG3 are different) in a coupling solvent in the presence of a coupling agent.

[0463] Embodiment 130. The method according to Embodiment 129, wherein the coupling agent is selected from the group consisting of (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), benzotriazolyloxy-tris[pyrrolidino]-phosphonium hexafluorophosphate (PyBOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP), hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT).

[0464] Embodiment 131. The method according to Embodiment 129, wherein the coupling agent comprises 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 1-hydroxy-7-azabenzotriazole (HOAT), or ethyl cyano(hydroxyimino)acetate (oxyma), diisopropylcarbodiimide (DIC), or dicyclohexylcarbodiimide (DCI).

[0465] Embodiment 132. The method according to any one of Embodiments 129 to 131, wherein the coupling solvent comprises dichloromethane or dimethylformamide.

[0466] Embodiment 133. The method according to Embodiments 129 to 132, wherein the coupling of the compound of Formula Y7 and Fmoc-D-Asp(OAll)-OH acid further comprises a base.

[0467] Embodiment 134. The method according to Embodiment 133, wherein the base comprises triethylamine or diisopropylethylamine.

[0468] Embodiment 135. The compound of Formula Y7 is of Formula Y8:

[0469] [Chemical Formula] (wherein PG4 is an amine protecting group) and is prepared by deprotecting in the presence of an amine deprotecting agent. The method according to any one of Embodiments 129 to 134.

[0470] Embodiment 136. PG4 is 4-methyltrityl, 4-methoxytrityl,

[0471] [Chemical Formula]

[0472] The method according to embodiment 135, wherein it is N-[1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)ethyl] (Dde), or 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene)-3-methylbutyl (ivDde).

[0473] Embodiment 137. The method according to embodiment 135, wherein PG4 is 4-methyltrityl and the deprotecting agent comprises hexafluoro-2-propanol, hydroxylamine, or hydrazine.

[0474] Embodiment 138. The method according to embodiment 135, wherein PG4 is 4-methyltrityl and the amine deprotecting agent is hexafluoro-2-propanol.

[0475] Embodiment 139. The compound of formula Y8 is a compound of formula Y9:

[0476]

Chemical formula

[0477]

Chemical formula

[0478] Embodiment 140. The method according to Embodiment 139, wherein the coupling agent is selected from the group consisting of (7-azabenzotriazol-1-yloxy) tripyrrolidinophosphonium hexafluorophosphate (PyAOP), benzotriazolyl-oxy-tris [pyrrolidino] - phosphonium hexafluorophosphate (PyBOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP), hexafluorophosphate azabenzotriazole tetramethyluroniumnium (HATU), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and 3- (diethoxyphosphoryloxy) -1,2,3-benzotriazin-4(3H)-one (DEPBT).

[0479] Embodiment 141. The method according to Embodiment 139, wherein the coupling agent comprises 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 1-hydroxy-7-azabenzotriazole (HOAT), or ethyl cyano(hydroxyimino)acetate (oxyma), diisopropylcarbodiimide (DIC), or dicyclohexylcarbodiimide (DCI).

[0480] Embodiment 142. The method according to any one of Embodiments 139 to 141, wherein the coupling solvent comprises dichloromethane or dimethylformamide.

[0481] Embodiment 143. The method according to Embodiments 139 to 142, wherein the coupling of the compound of formula Y7 and Fmoc-D-Asp(OAll)-OH acid further comprises a base.

[0482] Embodiment 144. The method according to Embodiment 143, wherein the base comprises triethylamine or diisopropylethylamine.

[0483] Embodiment 145. The following formula:

[0484]

Chemical formula

[0485] Embodiment 146

[0486]

Chemical Structure

[0487] Embodiment 147

[0488]

Chemical Structure

[0489] Embodiment 147. Formula V:

[0490]

Chemical Structure

[0491]

Chemical Structure

[0492]

Chemical Structure

[0493] Embodiment 148. The compound of formula V is hydrolyzed to give the following formula:

[0494] [Chemical formula] to form a compound of, the method according to embodiment 147, wherein the hydrolysis conditions depend on what the PG10 group is. When PG10 is a hydrolysable group, the hydrolysis conditions described herein can be used.

[0495] Embodiment 149. The compound of formula V is deprotected to give the following formula:

[0496] [Chemical formula] to form a compound of, the method according to embodiment 147.

[0497] The deprotection conditions used in this conversion depend on what the PG10 and PG11 groups are. For example, when PG11 is an Fmoc group, PG11 can be removed using the methods disclosed herein. Then, when PG10 is a tBu or another hydrolysable group, the tBu group and the resin can be removed using the hydrolysis methods disclosed herein.

[0498] One of ordinary skill in the art will appreciate that the invention described herein is not limited to that specifically shown and described. Rather, the scope of the invention is defined by the following claims. Further, it is to be understood that the foregoing description is merely illustrative of representative embodiments. This description is not intended to enumerate all possible variations. Alternative embodiments may not be presented for specific components of a composition or steps of a method, may result from different combinations of the described components, or other alternative embodiments not described may be available for a formulation, kit, or method and should not be regarded as a waiver of these alternative embodiments. It will be understood that many of these un-described embodiments are within the literal scope of the following claims and others are equivalent.

Claims

Claim 1 A method for preparing a compound of formula X: 【Chemical 1】 or a pharmaceutically acceptable salt thereof, said method comprising coupling a compound of formula Y: [Chemical Formula 2] wherein 【Chemical 3】 is a solid support with a compound of formula Z: [Chemical Formula 4] to form a compound of formula X: [Chemical Formula 5] Claim 2 The method according to claim 1, wherein the compound of formula X is treated with an acid in a hydrolyzing solvent to form a compound of formula A-1: Claim 3 【Chemical Formula 6】 The method according to claim 1 or 2, wherein the solid support comprises polystyrene cross-linked with divinylbenzene. Claim 4 The method according to any one of claims 1 to 3, wherein the coupling of the compounds of formula Y and formula Z comprises combining the compounds of formula Y and formula Z with a coupling agent in a coupling solvent. Claim 5 The method according to claim 4, wherein the coupling agent contains phosphorus. Claim 6 The method according to claim 4 or 5, wherein the coupling agent is selected from the group consisting of (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), benzotrizyloxy-tris[pyrrolidino]-phosphonium hexafluorophosphate (PyBOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP), hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT). Claim 7 The method according to claim 4, wherein the coupling agent comprises 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 1-hydroxy-7-azabenzotriazole (HOAT), or ethyl cyano(hydroxyimino)acetate (oxyma), diisopropylcarbodiimide (DIC), or dicyclohexylcarbodiimide (DCI). Claim 8 The method according to any one of claims 4 to 7, wherein the coupling solvent comprises dichloromethane or dimethylformamide. Claim 9 The method according to any one of claims 1 to 8, wherein the coupling of the compounds of formula Y and formula Z further comprises a base. Claim 10 ​ ​ The method according to claim 9, wherein the base comprises triethylamine or diisopropylethylamine.

11. The method according to any one of claims 2 to 10, wherein the acid comprises at least one of trifluoroacetic acid, hydrochloric acid, or para-toluenesulfonic acid, and the hydrolysis solvent comprises water.

12. The method according to claim 11, wherein the hydrolysis solvent further comprises at least one of triisopropylsilane or dithiothreitol.

13. The method according to any one of claims 2 to 12, wherein the acid comprises trifluoroacetic acid and the hydrolysis solvent comprises water.

14. The method according to any one of claims 2 to 12, wherein the acid comprises trifluoroacetic acid, the hydrolysis solvent comprises water, triisopropylsilane, and dithiothreitol, the coupling agent comprises PyAOP, the coupling solvent comprises dimethylformamide, and the coupling further comprises a base that is N,N-diisopropylethylamine.

15. The compound of formula A-2-R or a pharmaceutically acceptable salt thereof is prepared by treating the formula A-1 with a radioactive cation to form a compound of formula A-2-R: 【Chemical Formula 7】 (wherein Z is a radioactive cation) The method according to any one of claims 2 to 14.

16. Z is ４７ Sc, ５１ Cr, ５７ Co, ５８ Co, ５９ Fe, ６７ Cu, ６７ Ga, ６８ Ga, ７１ Ge, ７２ As, ７２ Se, ７７ As, ７７ Br, ８１ Rb, ８８ Y, ９０ Y, ９７ Ru, ９９ｍ Tc, １００ Pd, １０１ｍ Rh, １０３ Pb, １０５ Rh, １０９ Pd, １１１ Ag, １１１ In, １１３ In, １１９ Sb, １２１ Sn, １２７ Cs, １２８ Ba, １２９ Cs, １３１ Cs, １３１ I, １３９ La, １４０ La, １４２ Pr, １４３ Pr, １４９ Pm, １５１ Eu, １５３ Eu, １５３ Sm, １５９ Gr, １６１ Tb, １６５ Dy, １６６ Ho, １６９ Eu, １７５ Yb, １７７ Lu, １８６ Re, １８８ Re, １８９ Re, １９１ Os, １９３ Pt, １９４ Ir, １９７ Hg, １９８ Au, １９９ Ag, １９９ Au, ２０１ Tl, ２０３ Pb, ２１１ At, ２１２ Bi, ２１２ Pb, ２１３ Bi, ２２５ Ac, and ２２７ The method according to claim 15, selected from the group consisting of Th.

17. Z is １７７ Lu, ２２５ Ac, ２１１ At, ６７ Cu, １６１ Tb, ６７ Ga, ６８ Ga, ２０３ Pb, ２２３ Ra, and ２１２ The method according to claim 15, selected from the group consisting of Pb.

18. Z is ２２５ Ac, the method according to any one of claims 15 to 17.

19. Treating formula A-1 with a radioactive cation in an aqueous HCl solution ２２５ AcCl ３ The method according to claim 18, comprising combining a solution containing ２２５ , an aqueous buffer solution containing sodium ascorbate and ammonium acetate, and formula A-1.

20. The compound of the formula Y: 【Chemical 8】 is prepared by deprotecting a compound of formula Y2: 【Chemical Formula 9】 (wherein PG is an amine protecting group) The method according to any one of claims 1 to 14.

21. The method according to claim 20, wherein PG is selected from fluorenylmethoxycarbonyl (Fmoc), benzyl (Bn), and benzyloxycarbonyl (Cbz).

22. The method according to claim 20, wherein PG is fluorenylmethoxycarbonyl (Fmoc), and the deprotection of Y2 comprises treating Y2 with a base in an amine deprotection solvent.

23. The base is selected from the group consisting of cyclohexylamine, ethanolamine, piperidine, piperazine, triethylamine, and N,N-diisopropylethylamine, and the amine deprotection solvent contains at least one of dimethylformamide (DMF), dichloromethane, toluene, or N-methyl-2-pyrrolidone (NMP). The method according to claim 22.

24. The method according to claim 22, wherein the base contains piperidine and the amine deprotection solvent contains dimethylformamide.

25. The compound of formula Y2 is prepared by coupling a compound of formula Y3: 【Chemical 10】 with PG-D-Orn-OtBu (wherein PG is an amine protecting group). The method according to any one of claims 20 to 24.

26. The method according to claim 25, wherein the coupling occurs in a coupling solvent in the presence of a coupling agent.

27. The method according to claim 26, wherein the coupling agent contains phosphorus.

28. The coupling agent is selected from the group consisting of (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), benzotrizyloxy-tris[pyrrolidino]-phosphonium hexafluorophosphate (PyBOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP), hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT). The method according to claim 27.

29. The coupling agent contains 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 1-hydroxy-7-azabenzotriazole (HOAT), or ethyl cyano(hydroxyimino)acetate (oxyma), diisopropylcarbodiimide (DIC), or dicyclohexylcarbodiimide (DCI). The method according to claim 26.

30. The method according to any one of claims 26 to 29, wherein the coupling solvent contains dichloromethane or dimethylformamide.

31. The method according to any one of claims 25 to 29, wherein the coupling of the compound of formula Y3 and PG-D-Orn-OBu further comprises a base.

32. The method according to claim 31, wherein the base comprises triethylamine or diisopropylethylamine.

33. The compound of formula Y3 is prepared by deprotecting a compound of formula Y4: 【Chemical 11】 (wherein PG2 is a protecting group), and the deprotection is carried out in the presence of a carboxyl deprotecting agent and a carboxyl deprotecting solvent. The method according to any one of claims 25 to 32.

34. The method according to claim 33, wherein PG2 is selected from the group consisting of allyl, benzyl, and benzhydryl.

35. The method according to claim 33 or 34, wherein the carboxyl deprotecting solvent comprises at least one of dichloromethane, dimethylformamide, tetrahydrofuran, or ethanol.

36. The method according to any one of claims 33 to 35, wherein the carboxyl deprotecting solvent comprises dichloromethane and dimethylformamide.

37. The method according to any one of claims 33 to 36, wherein PG2 is allyl and the carboxyl deprotecting agent comprises Pd.

38. PG2 is allyl and the carboxyl deprotecting agent comprises Pd(PPh ３ ) ４ The method according to any one of claims 33 to 37.

39. PG2 is allyl and the carboxyl deprotecting agent contains Pd(PPh ３ ) ４ and the deprotection further includes a nucleophile. The method according to any one of claims 33 to 36.

40. The method according to claim 39, wherein the nucleophile is selected from the group consisting of 1,3-dimethylbarbituric acid and triphenylphosphine.

41. The compound of formula Y4 is prepared by coupling a compound of formula Y5: 【Chemical Formula 12】 with 3,5-bis(tert-butoxycarbonyl)benzoic acid in a coupling solvent in the presence of a coupling agent. The method according to any one of claims 33 to 40.

42. The method according to claim 41, wherein the coupling agent is selected from the group consisting of (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), benzotriazolyl-oxy-tris[pyrrolidino]-phosphonium hexafluorophosphate (PyBOP), bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP), hexafluorophosphate azabenzotriazole tetramethyluronium (HATU), hexafluorophosphate benzotriazole tetramethyluronium (HBTU), and 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT).

43. The method according to claim 41, wherein the coupling agent comprises 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 1-hydroxy-7-azabenzotriazole (HOAT), or ethyl cyano(hydroxyimino)acetate (oxyma), diisopropylcarbodiimide (DIC), or dicyclohexylcarbodiimide (DCI).

44. The method according to any one of claims 41 to 43, wherein the coupling solvent comprises dichloromethane or dimethylformamide.

45. The method according to any one of claims 41 to 44, wherein the coupling of the compound of formula Y5 and 3,5-bis(tert-butoxycarbonyl)benzoic acid further comprises a base.

46. The method according to claim 45, wherein the base comprises triethylamine or diisopropylethylamine.

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