Stable PSMA-targeting formulations

Abstract

The present disclosure relates to radiolysis stabilized pharmaceutical compositions that act as PSMA-modulators and methods of use thereof.

Description

[0001] LGPM Ref.: 773621: IST-001PC

[0002] STABLE PSMA-TARGETING FORMULATIONS

[0003] RELATED APPLICATIONS

[0004] This application claims priority to U.S. Provisional Application No. 63 / 755,549, filed February 7, 2025, the content of which is incorporated in its entirety.

[0005] BACKGROUND

[0006] Prostate cancer is the most prevalent type of cancer and the second leading cause of death from cancer in American men, with an estimated 179,000 cases and 37,000 deaths in 1999, (Landis, S. H. et al. CA Cancer J. Clin. 48:6-29 (1998)). The number of men diagnosed with prostate cancer is steadily increasing as a result of the increasing population of older men and a greater awareness of the disease leading to earlier diagnoses (Parker et al., 1997, CA Cancer J. Clin. 47:5-280). Prostate-specific membrane antigen (PSMA), a tumor-associated antigen and type II transmembrane protein, is expressed on the membrane of prostatic epithelial cells and overexpressed on prostate tumor cells as well as a variety of other solid tumors. It is a well-established target for nuclear imaging and therapy of prostate cancer. Prognosis is dependent on stage of the disease, and the five-year survival rate for advanced, disseminated prostate cancer is only 29% (Siegel R., et al. CA Cancer J Clin. 2012;62:220-41). Metastatic castration-resistant prostate cancer (mCRPC) is associated with poor prognosis and diminished quality of life. Treatment options for mCRPC patients include taxane-based therapies and second-line hormonal therapies, all showing moderate survival benefits. Unfortunately, those therapies are only temporarily effective, and development of treatment resistance is observed (Heidenreich A., et al. Nat Rev Urol.

[0007] 2014;11:189-90).

[0008] Radiolabeled small-molecule PSMA inhibitors are excellent candidates for prostate cancer therapies and diagnostics (“theranostics”) as they rapidly and efficiently localize in tumor lesions. PSMA l& T is a radiopharmaceutical agent comprising a PSMA-targeting ligand linked to a chelating agent that is chelated to a radioisotope. This radiopharmaceutical agent binds to PSMA-expressing tumor cells. Upon binding, PSMA-expressing tumor cells are either destroyed and / or imaged based on which radioisotope is used.

[0009] One of the major challenges in the production of radiopharmaceutical drugs however is to extend the shelf-life of product to allow treatment of patients at remote locations from the manufacturer site. As such, there remains a need for stable radiopharmaceutical formulations for cancer treatments.LGPM Ref.: 773621: IST-001PC

[0010] SUMMARY

[0011] Provided herein are radiolysis stabilized pharmaceutical compositions comprising a radiopharmaceutical agent and gelatin. The radiopharmaceutical agent can be a radiotherapeutic agent or a radioimaging agent. These radiolysis stabilized pharmaceutical compositions can be effective in treating and / or diagnosing cancer in a subject in need thereof as they exhibit superior radiochemical purity for long periods after production, which allows sufficient time to administer the composition to the subject.

[0012] As such, also provided herein are methods of treating and / or diagnosing cancer in a subject in need thereof comprising administering to the subject a radiolysis stabilized pharmaceutical composition of the present disclosure.

[0013] BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 depicts the negative effects on radiochemical purity due to ethanol in formulations of161Tb-PSMA l& T.

[0015] Figure 2 depicts the positive effects on radiochemical purity due to gelatin in formulations of161Tb-PSMA l& T.

[0016] Figure 3 depicts the positive effects on radiochemical purity due to gelatin in formulations of177Lu-PSMA l& T.

[0017] DETAILED DESCRIPTION

[0018] Recent developments in prostate cancer targeted radiopharmaceuticals have focused on PSMA ligands. PSMA is a favorable target for imaging and radionuclide therapy of prostate cancer, because it is overexpressed in 90-100% of local prostate cancer lesions, as well as in cancerous lymph node metastases and bone lesions. Furthermore, it has been shown that PSMA expression levels are further enhanced in high-grade, metastatic, and castration-resistant prostate cancer (Bostwick D. G., et al., Cancer. 1998;82:2256-61).

[0019] Numerous studies have shown the value of radiolabeled PSMA-targeted agents to visualize prostate cancer lesions, including prostate cancer metastases (Tagawa S. T., et al., Cancer.

[0020] 2010;116:1075-83; and Afshar-Oromieh A., et al., Eur J Nucl Med Mol Imaging.

[0021] 2015;42:197-209). Among the different PSMA-targeting tracers available, several tracers have been studied in clinical settings. These PSMA-targeting tracers have been used for imaging purposes using positron emission topography (PET) and single-photon emission computed tomography (SPECT) with radionuclides68Ga and111In, respectively (Afshar-Oromieh A., et al., 2015; and Budaus L., et al., Eur Urol. 2015). For therapeutic uses, radionuclides such as177Lu,90Y,131I, and213Bi have been explored (Weineisen M., et al., J Nucl Med. 2015;56:1169-76; and Maurer T., et al., Eur Urol. 2015;68:530-4). Theranostic compounds such as PSMA l& T are of great interest because they combine the potential ofLGPM Ref.: 773621: IST-001PC

[0022] diagnosis and radionuclide therapy using one PSMA targeting molecule (Weineisen M., et al., 2015).

[0023] While radioligand properties such as lipophilicity, target affinity, and metabolic stability, have been carefully optimized for theranostic applications, there remains a need for radiolysis stable formulations of such radioligand therapies. As radionuclides decay, a series of highly reactive chemicals are generated. In some cases, they can react with the drug substance, e.g., causing degradation of the radioisotope-containing drug and increasing radioactive impurity overtime.

[0024] During the phase I / II clinical trials, the radiolabeling process is typically performed inhouse at a hospital where the trial is conducted, just before injection to the patient. However, moving to a phase III study, and a commercial phase afterwards, radiopharmaceuticals need to be centrally manufactured and supplied to hundreds of patients in several centers across the US and Europe. To overcome potential stability limitations, especially radiolysis of the radiopharmaceutical post-production, radiolysis stabilized pharmaceutical compositions, which prolong the shelf-life of the product, are needed in order for it to be possible to transport the product to distant medical centers. Therefore, provided herein are radiolysis stabilized pharmaceutical compositions comprising a radiopharmaceutical agent and gelatin. As demonstrated in the Figures and the Examples, the inclusion of gelatin in the formulations described herein confers improved stability of the radiopharmaceutical agent, as measured by radiopharmaceutical purity.

[0025] Definitions

[0026] Listed below are definitions of various terms used to describe the compounds and compositions disclosed herein. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

[0027] Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art.

[0028] Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, and peptide chemistry are those well-known and commonly employed in the art.

[0029] As used in the specification and in the claims, the terms “comprise(s),” “comprising,” “include(s),’’ “having,” “has," “can,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that require the presence of the named features, groups, ingredients, or steps and does not exclude the presence of additional features, groups, ingredients, or steps. The term “comprise(s),” “comprising,”LGPM Ref.: 773621: IST-001PC

[0030] “include(s),” “having,” “has,” “can,” or “contain(s),” can include embodiments encompassed by the terms "consisting essentially of or "consisting of."

[0031] As used herein, the articles “a” and “an" refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element" means one element or more than one element. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.

[0032] As used herein, the term “about" will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. As used herein when referring to a measurable value such as an amount, a temporal duration, and the like, the term “about” is meant to encompass variations of ±10%, including ±5%, +4%, +3%, +2%, ±1%, and ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

[0033] As used herein, the term “essentially” means at the most fundamental level.

[0034] Therefore, the phrase “essentially free of,” in the context of the pharmaceutical compositions herein, means that the pharmaceutical composition comprises such minute amounts of an inert agent to have basically none of the inert agent present in the composition.

[0035] The term “administration” or the like as used herein refers to the providing a therapeutic agent to a subject. Multiple techniques of administering a therapeutic agent exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary, and topical administration.

[0036] The term “treat,” “treated," “treating," or “treatment” includes the diminishment or alleviation of at least one symptom associated or caused by the state, disorder or disease being treated. In certain embodiments, the treatment comprises alleviating the symptoms of prostate cancer.

[0037] As used herein, the term “prevent" or “prevention" means no disorder or disease development if none had occurred, or no further disorder or disease development if there had already been development of the disorder or disease. Also considered is the ability of one to prevent some or all of the symptoms associated with the disorder or disease.

[0038] As used herein, the term “patient," “individual,” or “subject” refers to a human or a non-human mammal. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and marine mammals. Preferably, the patient, subject, or individual is human.

[0039] As used herein, the terms “effective amount," “pharmaceutically effective amount,” and “therapeutically effective amount” refer to a nontoxic but sufficient amount of an agent to provide the desired biological result. That result may be reduction or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. AnLGPM Ref.: 773621: IST-001PC

[0040] appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation.

[0041] As used herein, “pharmaceutically acceptable” and “pharmaceutical grade” refer to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non-toxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0042] As used herein, the term “composition” or “pharmaceutical composition" refers to a mixture of at least one compound useful within the disclosure with a pharmaceutically acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a patient or subject. Multiple techniques of administering a compound exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary, and topical administration.

[0043] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the disclosure within or to the patient such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable" in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the disclosure, and not injurious to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations.

[0044] As used herein, “pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound useful within the present disclosure and are physiologically acceptable to the patient. Supplementary active compounds mayLGPM Ref.: 773621: IST-001PC

[0045] also be incorporated into the compositions. The “pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound disclosed herein. Other additional ingredients that may be included in the pharmaceutical compositions are known in the art and described, for example, in Remington’s Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.

[0046] As used herein, “radiochemical purity” and “RCP” is a measurement of radiolabeling efficiency where the activity of the radiolabeled peptide is compared to the activity of free isotope and / or activity of radiolysis products. RCP is measured by radio-high performance liquid chromatography (HPLC) and radio-thin layer chromatography (iTLC).

[0047] As used herein, “gelatin” or “gelatine” refers to a collection of peptides and proteins produced by partial hydrolysis of collagen (a structural protein in the extracellular matrix of the connective tissue of mammals) extracted from the skin, bones, and connective tissues of animals such as domesticated cattle, chicken, pigs, and fish. Gelatin is classified into two types: type A and type B. Type A gelatin is made from acid-treated collagen, usually derived from pigs, poultry, and fish, whereas type B gelatin is made from alkali-treated collagen, usually derived from bovine sources. Type AB gelatin is a mixture of both types A and B. See, e.g., Ward, A. G.; Courts, A. (1977). The Science and Technology of Gelatin. New York: Academic Press; The Gelatin Manufacturers Institute of America “Gelatin Handbook” 2017; and Fu, Y., et al., Critical Reviews in Food Science and Nutrition, 2018, 59(13), 2011-2027, the entire contents of which are incorporated herein.

[0048] “Hydrolyzed gelatin" or “hydrolyzed gelatine” refers to gelatin as described above, which has been subjected to further hydrolysis making it easier to dissolve in aqueous solutions. Hydrolyzed gelatin is usually considered to be non-gelling.

[0049] As used herein, “non-gelling gelatin” refers to gelatin having sufficiently low viscosity such that it is able to be administered via injection with a syringe without clogging said syringe or other administration device.

[0050] Pharmaceutical Compositions

[0051] Provided herein are radiolysis stabilized pharmaceutical compositions that are inhibitors and / or modulators of PSMA and are thus useful in the treatment of diseases and disorders, including cancer and other proliferation diseases.

[0052] In an aspect, provided herein is a radiolysis stabilized pharmaceutical composition comprising a radiopharmaceutical agent and gelatin.

[0053] In an embodiment, the radiopharmaceutical agent is a radiotherapeutic agent or a radioimaging agent. In another embodiment, the radiopharmaceutical agent is a PSMA binding ligand chelated or bound to a radioisotope.LGPM Ref.: 773621: IST-001PC

[0054] In an embodiment, the PSMA binding ligand chelated or bound to a radioisotope is selected from the group consisting of a ligand in Table 1, or any enantiomer or diastereomer thereof, wherein the radioisotope is not depicted.

[0055] Table 1

[0056] PSMA l& T (Zadavotide Guraxetan)

[0057]

[0058] LGPM Ref.: 773621: IST-001PC

[0059] (I-BA)KuE

[0060] HBED-CC-Ahx-KuE MIP-1404

[0061]

[0062] LGPM Ref.: 773621: IST-OO1PC

[0063] MIP-1405 MIP-1072 / MIP-1375 MIP-1095 / MIP-1466

[0064]

[0065] LGPM Ref.: 773621: IST-001PC

[0066]

[0067] LGPM Ref.: 773621: IST-001PC

[0068] DCFBC

[0069]

[0070] LGPM Ref.: 773621: IST-001PC

[0071] DCFPyL (Piflufolastat)

[0072] 0

[0073] HN HO2C ^—COaH

[0074] YC-88

[0075] HN \

[0076] HO2C '-~CO2H

[0077] PSMA-1007

[0078] PSMA5

[0079] CO2H

[0080]

[0081] LGPM Ref.: 773621: IST-001PC

[0082]

[0083] In an embodiment, the PSMA binding ligand chelated or bound to a radioisotope is selected from the group consisting of PSMA l& T, PSMA-11, and PSMA-1007. In another embodiment, the PSMA binding ligand chelated or bound to a radioisotope is PSMA l& T.LGPM Ref.: 773621: IST-001PC

[0084] Exemplary, but non-limiting, PSMA binding ligands chelated or bound to a radioisotope are shown in Table 2. In an embodiment, the PSMA binding ligand chelated or bound to a radioisotope is selected from the group consisting of a ligand in Table 2, or any enantiomer or diastereomer thereof.

[0085]

[0086] LGPM Ref.: 773621: IST-001PC

[0087]

[0088] In an embodiment, the radioisotope is an alpha-emitting isotope, in another embodiment, the radioisotope is a beta-emitting isotope. In another embodiment, the radioisotope is a gamma-emitting isotope. In another embodiment, the radioisotope is a positron-emitting isotope.

[0089] In another embodiment, the radioisotope is selected from the group consisting of225Ac,111Ag,211At,198Au,199Au,212Bi,213Bi,11C,55Co,57Co.64Cu,67Cu,165Dy,166Dy,18F,67Ga,68Ga,152Gd,153Gd,157Gd,157Gd,166Ho,123I,124I,131I,90In,111In,192lr,140La,177Lu,13N,15O,212Pb,103Pd,109Pd,149Pm,223Ra,224Ra,186Re,188Re,105Rh,43Sc,44Sc,47Sc,153Sm,

[0090]

[0091] 149Tb,151Tb,161Tb,94Tc,99mTc,227Th,86Y,90Y,169Yb,175Yb, and89Zr.

[0092] In another embodiment, the radioisotope is selected from the group consisting of151Tb,177Lu,18F, and68Ga. In an embodiment, the radioisotope is161Tb. In an embodiment, the radioisotope is177Lu. In an embodiment, the radioisotope is68Ga. In an embodiment, the pharmaceutical composition comprises161Dy.

[0093] In an embodiment, the gelatin is selected from the group consisting of type A gelatin, type B gelatin, type AB gelatin, and hydrolyzed gelatin. In another embodiment, the gelatin is hydrolyzed gelatin. In another embodiment, the gelatin is a non-gelling gelatin. In another embodiment, the gelatin is pharmaceutical grade gelatin.

[0094] In an embodiment, the pharmaceutical composition further comprises a buffer. In another embodiment, the buffer is selected from the group consisting of ascorbate, acetate, carbonate, citrate, histidine, phosphate, tartrate, and Tris buffers. In another embodiment, the buffer is an ascorbate buffer. In another embodiment, the buffer is an acetate buffer.

[0095] In another embodiment, the pH of the pharmaceutical composition at the end of production of the pharmaceutical composition is about 4 to about 5. In another embodiment, the pH of the pharmaceutical composition at the end of production of the pharmaceutical composition is about 4.5.

[0096] In an embodiment, the pharmaceutical composition further comprises saline.

[0097] In another embodiment, the pharmaceutical composition has at least about 99% radiochemical purity (RCP) by HPLC at the end of production of the pharmaceuticalLGPM Ref.: 773621: IST-001PC

[0098] composition. In an embodiment, the pharmaceutical composition has at least about 98% RCP by HPLC about 45 hours after production of the pharmaceutical composition. In an embodiment, the pharmaceutical composition has at least about 97% RCP by HPLC about 70 hours after production of the pharmaceutical composition. In an embodiment, the pharmaceutical composition has at least about 94% RCP by HPLC about 140 hours after production of the pharmaceutical composition.

[0099] In an embodiment, the pharmaceutical composition comprises:

[0100] a) PSMA l& T radiolabeled with a radioisotope;

[0101] b) gelatin;

[0102] c) a buffer; and

[0103] d) saline.

[0104] In an embodiment, the pharmaceutical composition comprises:

[0105] a) PSMA l& T radiolabeled with161Tb;

[0106] b) gelatin;

[0107] c) a buffer; and

[0108] d) saline.

[0109] In an embodiment, the pharmaceutical composition comprises:

[0110] a) PSMA l& T radiolabeled with a radioisotope;

[0111] b) hydrolyzed gelatin;

[0112] c) a buffer; and

[0113] d) saline.

[0114] In an embodiment, the pharmaceutical composition comprises:

[0115] a) PSMA l& T radiolabeled with a radioisotope;

[0116] b) gelatin;

[0117] c) an ascorbate buffer; and

[0118] d) saline.

[0119] In an embodiment, the pharmaceutical composition comprises:

[0120] a) PSMA l& T radiolabeled with161Tb;

[0121] b) hydrolyzed gelatin;

[0122] c) an ascorbate buffer; and

[0123] d) saline.

[0124] In an embodiment, the pharmaceutical composition has a ratio of PSMA l& T (pg) to161Tb (mCi) of about 0.5:1 to about 1:1.

[0125] In an embodiment, the pharmaceutical composition comprises:

[0126] a) PSMA l& T radiolabeled with177Lu;

[0127] b) gelatin;

[0128] c) a buffer; andLGPM Ref.: 773621: IST-001PC

[0129] d) saline.

[0130] In another embodiment, the pharmaceutical composition comprises: a) PSMA l& T radiolabeled with177Lu;

[0131] b) hydrolyzed gelatin;

[0132] c) a buffer; and

[0133] d) saline.

[0134] In another embodiment, the pharmaceutical composition comprises: a) PSMA l& T radiolabeled with177Lu;

[0135] b) hydrolyzed gelatin;

[0136] c) an ascorbate buffer; and

[0137] d) saline.

[0138] In an embodiment, the pharmaceutical composition comprises: a) PSMA-11 radiolabeled with68Ga;

[0139] b) gelatin;

[0140] c) a buffer; and

[0141] d) saline.

[0142] In another embodiment, the pharmaceutical composition comprises: a) PSMA-11 radiolabeled with68Ga;

[0143] b) hydrolyzed gelatin;

[0144] c) a buffer; and

[0145] d) saline.

[0146] In another embodiment, the pharmaceutical composition comprises: a) PSMA-11 radiolabeled with68Ga;

[0147] b) hydrolyzed gelatin;

[0148] c) an acetate buffer; and

[0149] d) saline.

[0150] In an embodiment, the pharmaceutical composition comprises: a) PSMA l& T chelated to161Dy;

[0151] b) gelatin;

[0152] c) a buffer; and

[0153] d) saline.

[0154] In another embodiment, the pharmaceutical composition comprises: a) PSMA l& T labeled with161Dy;

[0155] b) hydrolyzed gelatin;

[0156] c) an ascorbate buffer; and

[0157] d) saline.LGPM Ref.: 773621: IST-001PC

[0158] In an embodiment of the pharmaceutical compositions, the pharmaceutical composition comprises about 20 to about 100 mg of gelatin. In another embodiment, the pharmaceutical composition comprises about 30 to about 70 mg of gelatin. In another embodiment, the pharmaceutical composition comprises about 30 to about 50 mg of gelatin. In another embodiment, the pharmaceutical composition comprises about 35 mg of gelatin.

[0159] In an embodiment, the pharmaceutical composition is stable for about 2 years prior to radiolabeling. In an embodiment, the pharmaceutical composition is stable for about 3 hours to about 5 hours after radiolabeling. In an embodiment, the pharmaceutical composition is stable for about 4 hours after radiolabeling.

[0160] Methods of Treatment / Diagnosing / Imaging

[0161] In an aspect, provided herein is a method of treating cancer in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition of the present disclosure.

[0162] In an embodiment, the pharmaceutical composition comprises:

[0163] a) PSMA l& T radiolabeled with161Tb;

[0164] b) gelatin;

[0165] c) a buffer; and

[0166] d) saline.

[0167] In an embodiment, the pharmaceutical composition comprises:

[0168] a) PSMA l& T radiolabeled with161Tb;

[0169] b) hydrolyzed gelatin;

[0170] c) an ascorbate buffer; and

[0171] d) saline.

[0172] In an embodiment, provided herein is a method of treating cancer in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising:

[0173] a) PSMA l& T radiolabeled with177Lu;

[0174] b) gelatin;

[0175] c) a buffer; and

[0176] d) saline.

[0177] In another embodiment, the pharmaceutical composition comprises:

[0178] a) PSMA l& T radiolabeled with177Lu;

[0179] b) hydrolyzed gelatin;

[0180] c) a buffer; and

[0181] d) saline.

[0182] In another embodiment, the pharmaceutical composition comprises:LGPM Ref.: 773621: IST-001PC

[0183] a) PSMA l& T radiolabeled with177Lu;

[0184] b) hydrolyzed gelatin;

[0185] c) an ascorbate buffer; and

[0186] d) saline.

[0187] In an embodiment, the pharmaceutical composition of the present disclosure contains less than 1 mmol of sodium. In another embodiment, the pharmaceutical composition is essentially free of sodium.

[0188] In another aspect, provided herein is a method of diagnosing cancer in a subject comprising administering to the subject a pharmaceutical composition of the present disclosure.

[0189] In another aspect, provided herein is a method of diagnosing or treating a disease or disorder in a subject in need thereof comprising administering to the subject an effective amount of a pharmaceutical composition of the present disclosure. In an embodiment, the disease or disorder is cancer. In an embodiment, the disease or disorder is cancer associated with PSMA activity. In another embodiment, the disease or disorder is prostate cancer. In yet another embodiment, the disease or disorder is metastatic prostate cancer. In still another embodiment, the disease or disorder is high-grade prostate cancer. In another embodiment, the disease or disorder is castration-resistant prostate cancer prostate cancer.

[0190] In an embodiment, the methods discussed herein further comprise conducting positron-emission tomography on the subject following administration of the pharmaceutical composition.

[0191] One aspect of this disclosure provides pharmaceutical compositions that are useful for the treatment and / or diagnosis of diseases, disorders, and conditions characterized by excessive or abnormal cell proliferation. Such diseases include, but are not limited to, a proliferative or hyperproliferative disease, and a neurodegenerative disease. Examples of proliferative and hyperproliferative diseases include, without limitation, cancer.

[0192] In an embodiment, the cancer is associated with PSMA activity. In another embodiment, the cancer is prostate cancer. In another embodiment, the cancer is metastatic prostate cancer. In yet another embodiment, the disease or disorder is high-grade prostate cancer. In still another embodiment, the disease or disorder is castration-resistant prostate cancer prostate cancer.

[0193] In an embodiment, the cancer is selected from the group consisting of prostate, breast, ovary, cervix, testis, genitourinary tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung adenocarcinoma, bone, colon, colorectal, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder carcinoma, liver carcinomaLGPM Ref.: 773621: IST-001PC

[0194] and biliary passages, kidney carcinoma, myeloid disorders, lymphoid disorders, Hodgkin's, hairy cells, buccal cavity and pharynx (oral), lip, tongue, mouth, pharynx, small intestine, colon, rectum, large intestine, rectum, brain and central nervous system, chronic myeloid leukemia (CML), and leukemia.

[0195] In an embodiment of the methods, the subject is a human.

[0196] In another embodiment, the cancer is selected from the group consisting of myeloma, lymphoma, or a cancer selected from gastric, renal, head and neck, oropharangeal, nonsmall cell lung cancer (NSCLC), endometrial, hepatocarcinoma, non-Hodgkin’s lymphoma, and pulmonary.

[0197] In an embodiment, the cancer is selected from the group consisting of prostate cancer, colon cancer, lung cancer, squamous cell cancer of the head and neck, esophageal cancer, hepatocellular carcinoma, melanoma, sarcoma, gastric cancer, pancreatic cancer, ovarian cancer, breast cancer.

[0198] In an embodiment, the cancer is selected from the group consisting of tumors, neoplasms, carcinomas, sarcomas, leukemias, lymphomas and the like. For example, cancers include, but are not limited to, mesothelioma, leukemias and lymphomas such as cutaneous T-cell lymphomas (CTCL), noncutaneous peripheral T-cell lymphomas, lymphomas associated with human T-cell lymphotrophic virus (HTLV) such as adult T-cell leukemia / lymphoma (ATLL), B-cell lymphoma, acute nonlymphocytic leukemias, chronic lymphocytic leukemia, chronic myelogenous leukemia, acute myelogenous leukemia, lymphomas, and multiple myeloma, non-Hodgkin lymphoma, acute lymphatic leukemia (ALL), chronic lymphatic leukemia (CLL), Hodgkin's lymphoma, Burkitt lymphoma, adult T-cell leukemia lymphoma, acute-myeloid leukemia (AML), chronic myeloid leukemia (CML), or hepatocellular carcinoma. Further examples include myelodysplastic syndrome, childhood solid tumors such as brain tumors, neuroblastoma, retinoblastoma, Wilms' tumor, bone tumors, and soft-tissue sarcomas, common solid tumors of adults such as head and neck cancers (e.g., oral, laryngeal, nasopharyngeal and esophageal), genitourinary cancers (e.g., prostate, bladder, renal, uterine, ovarian, testicular), lung cancer (e.g., small-cell and nonsmall cell), breast cancer, pancreatic cancer, melanoma and other skin cancers, stomach cancer, brain tumors, tumors related to Gorlin syndrome (e.g., medulloblastoma, meningioma, etc.), and liver cancer. Additional exemplary forms of cancer which may be treated by the subject compounds include, but are not limited to, cancer of skeletal or smooth muscle, stomach cancer, cancer of the small intestine, rectum carcinoma, cancer of the salivary gland, endometrial cancer, adrenal cancer, anal cancer, rectal cancer, parathyroid cancer, and pituitary cancer.

[0199] Additional cancers that the compounds described herein may be useful in treating are, for example, colon carcinoma, familial adenomatous polyposis carcinoma andLGPM Ref.: 773621: IST-001PC

[0200] hereditary non-polyposis colorectal cancer, or melanoma. Further, cancers include, but are not limited to, labial carcinoma, larynx carcinoma, hypopharynx carcinoma, tongue carcinoma, salivary gland carcinoma, gastric carcinoma, adenocarcinoma, thyroid cancer (medullary and papillary thyroid carcinoma), renal carcinoma, kidney parenchyma carcinoma, cervix carcinoma, uterine corpus carcinoma, endometrium carcinoma, chorion carcinoma, testis carcinoma, urinary carcinoma, melanoma, brain tumors such as glioblastoma, astrocytoma, meningioma, medulloblastoma and peripheral neuroectodermal tumors, gall bladder carcinoma, bronchial carcinoma, multiple myeloma, basalioma, teratoma, retinoblastoma, choroidea melanoma, seminoma, rhabdomyosarcoma, craniopharyngeoma, osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma, Ewing sarcoma, and plasmocytoma.

[0201] In another aspect, provided herein is the use of one or more compounds of the disclosure in the manufacture of a medicament for the treatment of cancer, including without limitation the various types of cancer disclosed herein.

[0202] In some embodiments, the compounds of this disclosure are useful for treating cancer, such as colorectal, thyroid, breast, and lung cancer; and myeloproliferative disorders, such as polycythemia vera, thrombocythemia, myeloid metaplasia with myelofibrosis, chronic myelogenous leukemia, chronic myelomonocytic leukemia, hypereosinophilic syndrome, juvenile myelomonocytic leukemia, and systemic mast cell disease. In some embodiments, the compounds of this disclosure are useful for treating hematopoietic disorders, in particular, acute-myelogenous leukemia (AML), chronic-myelogenous leukemia (CML), acute-promyelocytic leukemia, and acute lymphocytic leukemia (ALL).

[0203] In an aspect, provided herein is the use of a pharmaceutical composition of present disclosure as an imaging agent. In an embodiment, the imaging agent is used in positron emission tomography (PET). In an embodiment, the imaging agent is used in PET about 50 minutes to about 100 minutes after administration to the subject. In an embodiment, the imaging agent is used in PET about 60 minutes after administration to the subject.

[0204] In another embodiment, the imaging agent is used in single-photon emission computed tomography (SPECT). In another embodiment, the imaging agent is used to visualize cell death.

[0205] In an embodiment, the imaging agent comprises:

[0206] a) PSMA l& T radiolabeled with68Ga;

[0207] b) gelatin;

[0208] c) a buffer; and

[0209] d) saline.

[0210] In another embodiment, the imaging agent comprises:LGPM Ref.: 773621: IST-001PC

[0211] a) PSMA l& T radiolabeled with68Ga;

[0212] b) hydrolyzed gelatin;

[0213] c) a buffer; and

[0214] d) saline.

[0215] In another embodiment, the imaging agent comprises:

[0216] a) PSMA l& T radiolabeled with68Ga;

[0217] b) hydrolyzed gelatin;

[0218] c) a acetate buffer; and

[0219] d) saline.

[0220] In an embodiment, the imaging agent comprises:

[0221] a) PSMA-11 radiolabeled with68Ga;

[0222] b) gelatin;

[0223] c) a buffer; and

[0224] d) saline.

[0225] In another embodiment, the imaging agent comprises:

[0226] a) PSMA-11 radiolabeled with68Ga;

[0227] b) hydrolyzed gelatin;

[0228] c) a buffer; and

[0229] d) saline.

[0230] In another embodiment, the imaging agent comprises:

[0231] a) PSMA-11 radiolabeled with68Ga;

[0232] b) hydrolyzed gelatin;

[0233] c) an acetate buffer; and

[0234] d) saline.

[0235] In an embodiment, the imaging agent comprises:

[0236] a) PSMA-1007 radiolabeled with18F;

[0237] b) gelatin;

[0238] c) a buffer; and

[0239] d) saline.

[0240] In another embodiment, the imaging agent comprises:

[0241] a) PSMA-1007 radiolabeled with18F;

[0242] b) hydrolyzed gelatin;

[0243] c) a buffer; and

[0244] d) saline.

[0245] In another embodiment, the imaging agent comprises:

[0246] a) PSMA-1007 radiolabeled with18F;

[0247] b) hydrolyzed gelatin;LGPM Ref.: 773621: IST-001PC

[0248] c) an ascorbate buffer; and

[0249] d) saline.

[0250] In an embodiment of the methods and diagnosing, the pharmaceutical composition is administered at a dose of about 1 MBq / kg to about 3 MBq / kg wherein “per kilogram” ( / kg) refers to the body weight of the subject. In another embodiment, the pharmaceutical composition is administered at a dose of about 1.5 MBq / kg to about 2.5 MBq / kg. In another embodiment, the pharmaceutical composition is administered at a dose of about 1.8 MBq / kg to about 2.2 MBq / kg.

[0251] In another embodiment, the pharmaceutical composition is administered at a dose of about 100 MBq to about 300 MBq. In another embodiment, the pharmaceutical composition is administered at a dose of about 100 MBq to about 280 MBq. In another embodiment, the pharmaceutical composition is administered at a dose of about 111 MBq (3 mCi) to about 259 MBq (7 mCi).

[0252] In another embodiment, the pharmaceutical composition is administered at a dose of about 7 mCi. In another embodiment, the pharmaceutical composition is administered at a dose less than 7 mCi.

[0253] In another embodiment, the pharmaceutical composition comprises about 50 mCi to about 100 mCi radioactivity. In another embodiment, the pharmaceutical composition comprises about 60 mCi to about 80 mCi radioactivity. In another embodiment, the pharmaceutical composition comprises about 70 mCi radioactivity. In another embodiment, the pharmaceutical composition comprises about five doses.

[0254] In an embodiment, the pharmaceutical composition comprises not more than 10 μg of the radiopharmaceutical agent per dose. In an embodiment, the pharmaceutical composition comprises less than 10 μg of the radiopharmaceutical agent per dose. In an embodiment, the pharmaceutical composition comprises about 10 μg of the radiopharmaceutical agent per dose.

[0255] In another embodiment, the pharmaceutical composition is administered intravenously. In another embodiment, the pharmaceutical composition is administered in a single injection. In another embodiment, the pharmaceutical composition is administered slowly via a single intravenous injection. In another embodiment, an aqueous sodium chloride solution is administered to the subject after administration of the pharmaceutical composition. In another embodiment, the aqueous sodium chloride solution is a 9 mg / mL (0.9%) NaCI solution.

[0256] In an embodiment of the methods, the pharmaceutical composition or imaging agent is administered intravenously. In another embodiment of the methods, the pharmaceutical composition or imaging agent is administered by infusion.LGPM Ref.: 773621: IST-001PC

[0257] In another embodiment, the pH of the pharmaceutical composition or imaging agent at the time of administrating the pharmaceutical composition or imaging agent is about 4 to about 8. In another embodiment, the pH of the pharmaceutical composition or imaging agent at the time of administrating the pharmaceutical composition or imaging agent is about 7.

[0258] Due to its radioactivity, the compositions provided herein are packaged using internationally accepted radiation protectant packaging materials, e.g., Type A packages. Shipping from the production site is undertaken from 1 to 24 hours after the end of production, once packaging is completed and shipping is authorized.

[0259] Therefore, in an embodiment, the compositions of the present disclosure are received at the site of administration to a human subject up to 96-120 hours after end of production.

[0260] The compositions are removed from their protective container under standard radiation safety procedures. The compositions are administered intravenously as an injection using a disposable syringe and a syringe shield, as an infusion using the gravity method, or as an infusion using the vial.

[0261] Administration / Dosages / Formulations

[0262] Injectable preparations (for example, sterile, injectable aqueous or oleaginous suspensions) may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U. S. P., and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0263] The pharmaceutical compositions of the present disclosure can be administered intravenously or by infusion (IV). In an embodiment, the pharmaceutical compositions of the present disclosure are administered intravenously as an injection using a disposable syringe and a syringe shield, as an infusion using the gravity method, or as an infusion using a vial. In an embodiment, the compositions herein are administered to the human subject by slow intravenous injection over a duration of approximately 1 to 10 minutes. In another embodiment, the area of injection is flushed before and after administration of the present pharmaceutical compositions using 5 to 20 mL of 0.9% sterile saline. In another embodiment, the present compositions are administered to the human subject by IV injection within 144 hours after the end of production.LGPM Ref.: 773621: IST-001PC

[0264] The pharmaceutical compositions of the present disclosure for use in a subject (e.g., human) are typically administered by IV at a therapeutic dose or a dose sufficient for imaging. When administered intravenously via infusion, the dosage may depend upon the infusion rate at which an IV formulation is administered. In general, the therapeutically effective dosage of a pharmaceutical composition is dependent on the species of the subject, the body weight, age and individual condition, the disorder or disease or the severity thereof being treated.

[0265] According to the methods of treatment of the present disclosure, disorders are treated in a subject, such as a human or other animal, by administering to the subject a therapeutically effective amount of a pharmaceutical composition of the disclosure, in such amounts and for such time as is necessary to achieve the desired result. The term “therapeutically effective amount” of a pharmaceutical composition of the disclosure, as used herein, means a sufficient amount of the pharmaceutical composition so as to decrease the symptoms of a disorder in a subject. As is well understood in the medical arts, a therapeutically effective amount of a pharmaceutical composition of this disclosure will be at a reasonable benefit / risk ratio applicable to any medical treatment.

[0266] In general, pharmaceutical composition of the disclosure will be administered in therapeutically effective amounts via any of the usual and acceptable modes known in the art, either singly or in combination with one or more therapeutic agents. A therapeutically effective amount may vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used and other factors.

[0267] Upon improvement of a subject’s condition, a maintenance dose of a pharmaceutical composition of this disclosure may be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained; when the symptoms have been alleviated to the desired level, treatment should cease. The subject may, however, require intermittent treatment on a long-term basis upon any recurrence of disease symptoms.

[0268] It will be understood, however, that the total daily usage of the compositions of the present disclosure will be decided by the attending physician within the scope of sound medical judgment. The specific inhibitory dose for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts.LGPM Ref.: 773621: IST-001PC

[0269] The disclosure also provides for a pharmaceutical combination, e.g., a kit, comprising a) a first agent which is a compound of the disclosure as disclosed herein, in free form or in pharmaceutically acceptable salt form, and b) at least one co-agent. The kit can comprise instructions for its administration.

[0270] Some examples of materials which can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers; alumina; aluminum stearate; lecithin; serum proteins, such as human serum albumin; buffer substances such as phosphates, glycine, sorbic acid, or potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes, such as protamine sulfate; disodium hydrogen phosphate; potassium hydrogen phosphate; sodium chloride; zinc salts; colloidal silica; magnesium trisilicate; polyvinyl pyrrolidone; polyacrylates; waxes; polyethylenepolyoxypropylene-block polymers; wool fat; sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such a propylene glycol or polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; and phosphate buffer solutions. Further, non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator. The protein kinase inhibitors or pharmaceutical salts thereof may be formulated into pharmaceutical compositions for administration to animals or humans. These pharmaceutical compositions, which comprise an amount of the protein inhibitor effective to treat or prevent a protein kinase-mediated condition and a pharmaceutically acceptable carrier, are other embodiments of the present disclosure.

[0271] Kits

[0272] In an aspect, provided herein is a kit comprising a pharmaceutical composition of the present disclosure. The kit can comprise instructions for administering the pharmaceutical composition for treating or imaging cancer in a subject. The kit can comprise instructions for preparing the pharmaceutical composition via radiolabeling.

[0273] In another aspect, the disclosure provides a kit comprising a pharmaceutical composition capable of modulating PSMA activity.LGPM Ref.: 773621: IST-001PC

[0274] In yet another aspect, provided herein is a kit comprising a pharmaceutical composition disclosed herein for the treatment of any of the indications disclosed herein.

[0275] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents were considered to be within the scope of this disclosure and covered by the claims appended hereto. For example, it should be understood, that modifications in reaction conditions, including but not limited to reaction times, reaction size / volume, and experimental reagents, such as solvents, catalysts, pressures, atmospheric conditions, e.g., nitrogen atmosphere, and reducing / oxidizing agents, with art-recognized alternatives and using no more than routine experimentation, are within the scope of the present application.

[0276] It is to be understood that wherever values and ranges are provided herein, all values and ranges encompassed by these values and ranges, are meant to be encompassed within the scope of the present disclosure. Moreover, all values that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the present application.

[0277] The following examples further illustrate aspects of the present disclosure. However, they are in no way a limitation of the teachings of the present disclosure as set forth.

[0278] EXAMPLES

[0279] The pharmaceutical compositions and methods disclosed herein are further illustrated by the following examples, which should not be construed as further limiting. The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of organic synthesis, radiopharmaceuticals, and formulations, which are within the skill of the art.

[0280] The radiolabeled pharmaceutical compositions provided herein were prepared on a Trasis module following the process described in “Automated Synthesis of 68Ga / 177Lu-PSMA on the Trasis miniAllinOne", J Labelled Comp Radiopharm. 2020 Jun 30;63(8):393-403, the content of which is incorporated by reference. These pharmaceutical compositions include, but are not limited to, compositions comprising PSMA l& T radiolabeled with161Tb, PSMA l& T radiolabeled with177Lu, or PSMA-1007 radiolabeled with18F. Radiolabeled pharmaceutical compositions comprising PSMA-11 radiolabeled with68Ga were prepared using the cold kit, Isoprotrace®.

[0281] The pharmaceutical compositions were prepared with various ratios of the precursor’s (PSMA binding ligand) mass (pg) to the activity of the radioisotope (mCi). The ratio of PSMA l& T to 177-Lu and 161-Tb were in the range of 0.5:1 to 1:1; the ratio of PSMA-11 to Ga-68 were in the range of about 1:3 to 1:6; and the ratio of PSMA-1007 to F-18 wereLGPM Ref.: 773621: IST-001PC

[0282] in the range of about 0.5:1 to 1.6:1. Pharmaceutical compositions comprising PSMA binding ligands radiolabeled to68Ga comprise an acetate buffer whereas all other pharmaceutical compositions comprise an ascorbate buffer.

[0283] The hydrolyzed gelatin in the pharmaceutical compositions of the present disclosure was purchased from commercial sources wherein the hydrolyzed gelatin is intended for biomedical applications.

[0284] Example 1: Evaluating the effect of ethanol for maintaining RCP

[0285] A composition comprising PSMA l& T radiolabeled with161Tb, ascorbate buffer, and saline was prepared in order to determine the effect of ethanol, a known radioprotectant, on RCP over time. The preparation of this composition was accomplished using the following precursors:

[0286] Material Additional info

[0287] Tb-161 Vol. 1.25 mL

[0288]

[0289] PSMA l& T sol. 250 μg / 750 μL

[0290] Stability

[0291] The solution’s RCP was tested at the End of Production (EOP) and over time with a reduction in RCP already measured at day 1. At this stage, the solution was subdivided into two vials: one vial remained as is, while 10% V / V ethanol was added to the second vial in attempt to prevent or slow radiolysis. Samples of the two vials were tested for RCP (by HPLC) and the results are provided in Table 3 and shown in Figure 1.

[0292] Table 3

[0293] Days from EOP RCP (by HPLC) RCP (by HPLC)

[0294] with ethanol without ethanol

[0295] 0 (at EOP) 100%

[0296] 1 98.36%

[0297] 2 97.61% 97.77%

[0298] 3 97.12% 97.06%

[0299]

[0300] 4 96.53% 96.93%

[0301] As can be seen, ethanol did not improve RCP or slow radiolysis despite being considered a radioprotectant.

[0302] Example 2: Evaluating the effect of gelatin for maintaining RCP

[0303] The pharmaceutical composition described in Example 1 was prepared with the following modifications: the ratio of the precursor’s mass (pg) to the activity of the radioisotope (mCi) was changed to 0.5:1. The synthesis was accomplished using the following precursors:

[0304] Material Amount Additional info

[0305]

[0306] Tb-161 125 mCi Vol. 0.545 mLLGPM Ref.: 773621: IST-001PC

[0307] PSMA l& T sol. 190 μl (containing 63μg of

[0308]

[0309] (250 μg / 750 μL) the peptide)

[0310] All excipients used were the same as those used in the Trasis module. At the end of the process, a sample of the product was tested for RCP by iTLC and HPLC. Both tests resulted in 100% RCP.

[0311] Stability

[0312] Once the efficiency of the radiolabeling was determined (at EOP), the solution was subdivided into two vials (8.5 mL in each vial): one vial remained as is, while 1 mL of a gelatin solution (containing 34.8 mg of hydrolyzed gelatin) was added to the second vial.

[0313] Samples of the two vials were tested for RCP (by HPLC) and the results are provided in Table 4 and shown in Figure 2.

[0314] Table 4

[0315] Time (hrs) RCP (by HPLC)

[0316] elapsed from EOP161Tb-PSMA161Tb-PSMA

[0317] with gelatin without gelatin

[0318] 0 (at EOP) 100% 100%

[0319] 21 100% 100%

[0320] 45 100% 100%

[0321] 69 99% 96%

[0322]

[0323] 141 95.4% 91.6%

[0324] As can be seen, PSMA l& T can be efficiently radiolabeled with Tb-161 even when applying a ratio of 0.5:1 peptide (pg) to Tb-161 (mCi). The stability of the sample containing gelatin is better than that of the sample of same lot tested without gelatin.

[0325] The stabilizing effect of gelatin is further evidenced by the RCP values measured for pharmaceutical compositions comprising PSMA l& T radiolabeled with177Lu, ascorbate buffer, and saline with or without hydrolyzed gelatin. These results are shown in Table 5 and Figure 3.

[0326] Table 5

[0327] Time (hrs) RCP (by HPLC)

[0328] elapsed from177Lu-PSMA l& T177Lu-PSMA l& T

[0329] EOP with gelatin without gelatin

[0330] 0 (at EOP) 100% 100%

[0331] 19 100% 100%

[0332] 96 97% 87%

[0333] 114 96.2% 86.7%

[0334]

[0335] 138 94.7% 82.6%

[0336] The results indicate that PSMA l& T can be efficiently radiolabeled using an automated synthesis module and commercially available consumables. While the RCP ofLGPM Ref.: 773621: IST-001PC

[0337] the product decreased over time in all experiments, the addition of hydrolyzed gelatin had a beneficial effect on the stability of the Tb-161 and Lu-177 labelled PSMA l& T.

[0338] Example 3: Clinical Diagnosis

[0339] • A pharmaceutical composition comprising gozetotide trifluoroacetate equivalent to 10 micrograms of gozetotide (PSMA-11 ) radiolabeled with gallium (68Ga) is indicated for positron emission tomography (PET) of prostate-specific membrane antigen (PSMA)-positive lesions in men with prostate cancer in the following clinical settings: Primary staging of patients with high risk prostate cancer prior to primary curative therapy.

[0340] • Suspected prostate cancer recurrence based on elevated serum prostatespecific antigen (PSA) level, after primary curative therapy.

[0341] The recommended dose of gallium (68Ga) gozetotide is 1.8-2.2 MBq / kg of body weight, with a minimum dose of 111 MBq up to a maximum dose of 259 MBq.

[0342] Each kit for the preparation of gallium (68Ga) gozetotide injection contains multi-dose vials in a carton and package insert.

[0343] Each multi-dose vial consists of 10 pg gozetotide (active pharmaceutical ingredient (API) precursor) as a white to off-white freeze-dried plug or powder in a 10 mL colorless Type I glass vial with a rubber stopper and aluminum flip-off cap. After radiolabeling with gallium (68Ga) chloride solution obtained from a commercial source, the vial contains a sterile, non-pyrogenic gallium (68Ga) gozetotide injection.

[0344] In radiopharmacies, the vial is reconstituted with 1.1 mL to 5 mL of gallium (68Ga) chloride solution containing up to 70 mCi of radioactivity (the reconstitution procedure is provided below). After reconstitution, the vial is swirled, inverted, and placed upright. The vial is then incubated at room temperature between 20 °C to 30 °C (68 °F to 86 °F) for at least five minutes without agitation or stirring in upright orientation.

[0345] After radiolabeling, gallium (68Ga) gozetotide solution for injection may be diluted with sterile water for injection, USP or 0.9% sodium chloride injection, USP up to a final volume of 10mL.

[0346] Composition of Gallium t^Ga) Gozetotide Drug Product

[0347]

[0348] The qualitative and quantitative composition of the drug product kit for the preparation of gallium (68Ga) gozetotide injection (prior to lyophilization) is presented in Table 6.LGPM Ref.: 773621: IST-001PC

[0349] The density of the bulk gozetotide solution is 1.043 g / mL and the fill volume of the drug product in a multi-dose vial is 1 mL. The percentage of each ingredient in the multidose vial prior to lyophilization was calculated on the weight per weight basis (% w / w), considering the bulk density of 1.043 g / mL.

[0350] Table 6

[0351] Composition

[0352] Quantity per unit

[0353] Ingredients % w / w (mg / vial)

[0354] Active Ingredient

[0355] Gozetotide 0.01-0.0310.0009%

[0356] Excipients

[0357] Sodium acetate anhydrous 62.0 5.9%

[0358] Gelatin

[0359] 34.8 3.3% (hydrolyzed, non-gelling)

[0360] Sodium chloride 6.3 0.6%

[0361] Q. S. to 1.0 mL

[0362] Sterile water for injection290.1%

[0363] (940 mg)3

[0364] Total theoretical weight 10434100%

[0365]

[0366] Q. S. = Quantity sufficient

[0367] 1On anhydrous TFA-free base

[0368] 2Solvent used prior to lyophilization of 1.0 mL of bulk gozetotide solution. Removed during the lyophilization cycle and traces will be present in freeze-dried cake.

[0369] 31043 mg - (0.01 mg + 62 mg + 34.8 mg + 6.3 mg) = 940 mg

[0370] 4Total quantity filled in vials before lyophilization = 1 mL, which corresponds to 1043 mg based on the density of the bulk solution.

[0371] Method of administration

[0372] This medicinal product is administered intravenously as a single injection. It is reconstituted and radiolabeled before administration to the patient. After reconstitution and radiolabeling, gallium (68Ga) gozetotide solution is administered by slow intravenous injection. Local extravasation resulting in inadvertent radiation exposure to the patient and imaging artefacts should be avoided. The injection is followed by an intravenous flush of sodium chloride 9 mg / mL (0.9%) solution for injection to ensure full delivery of the dose.

[0373] The total radioactivity in the syringe is verified with a dose calibrator immediately before and after administration to the patient. The dose calibrator is calibrated and comply with international standards.

[0374] Image acquisition

[0375] Gallium (68Ga) gozetotide is suitable for PET medical imaging. Patients are encouraged to void immediately prior to image acquisition. The patient is positioned supine with the arms above the head, as tolerated by the patient. A CT scan is obtained forLGPM Ref.: 773621: IST-001PC

[0376] attenuation correction and anatomical correlation. The acquisition includes a whole-body acquisition from the base of the skull to mid-thigh.

[0377] PET images are acquired 50 to 100 (ideally 60) minutes after the intravenous administration of gallium (68Ga) gozetotide solution. Imaging acquisition start time and duration is adapted according to the equipment used, the patient, and the tumor characteristics in order to obtain the best image quality possible.

[0378] Patient preparation

[0379] Patients are not required to fast. Patients are allowed to take all their medications. PSMA-expression may be increased by androgen-deprivation therapy, but the clinical significance is unclear. The patient should be well-hydrated before the start of the examination and is urged to void just prior to the image acquisition. The patient is urged to void as often as possible during the first hours after the examination in order to reduce radiation. Concomitant administration of loop diuretics can lower the activity of gallium (68Ga) gozetotide in the bladder and ureter and decrease peri-bladder artefacts.

[0380] Specific warnings

[0381] This medicinal product contains less than 1 mmol sodium (23 mg) per vial, that is to say essentially “sodium-free.”

[0382] Undesirable effects

[0383] Exposure to ionizing radiation is linked with cancer induction and a potential for development of hereditary defects. As the effective dose, resulting from the administration of an average activity of 2 MBq / kg to an 80 kg man is 3.5 mSv, these adverse reactions are expected to occur with a low probability. Mild to moderate adverse reactions occurred in patients receiving gallium (68Ga) gozetotide. The most commonly reported reactions were fatigue, headache, injection site reactions, nausea, and rash.

[0384] Pharmacodynamic properties

[0385] Mechanism of action

[0386] Gallium (68Ga) gozetotide binds to prostate-specific membrane antigen (PSMA). It binds to cells that express PSMA, including malignant prostate cancer cells, which usually overexpress PSMA. Gallium (68Ga) is a p+ emitting radionuclide that allows positron emission tomography (PET).LGPM Ref.: 773621: IST-001PC

[0387] Half-life

[0388] Based on the gallium (68Ga) gozetotide biological and terminal half-life of 4.4 hours and on the gallium (68Ga) physical half-life of 68 minutes, the resulting gallium (68Ga) gozetotide effective half-life is 54 minutes.

[0389] Incompatibilities

[0390] Radiolabeling of carrier molecules with gallium (68Ga) chloride is very sensitive to the presence of trace metal impurities. Only syringe and syringe needles able to minimize trace metal impurity levels (for example, non-metallic or coated with silicone needles) are used.

[0391] Preparation for multidose application

[0392] The medicinal product is supplied as a single-vial kit. Preparation of gallium (68Ga) gozetotide solution is to be done according to the following aseptic procedure:

[0393] For reconstitution and radiolabeling with Eckert & Ziegler GalliaPharm Generator or Curium GalenVita-.

[0394] a. Before use, bring the product vial to room temperature for a minimum of 10 minutes.

[0395] b. Remove the cap from the top of the vial and sanitize the closure of the vial with an appropriate alcohol pad to disinfect the surface and allow to air dry.

[0396] c. Place the vial in a suitable shielding.

[0397] d. Connect a 0.2 μm vent filter to a short sterile needle (e.g., 5 / 8”-1”) and inject the vial. Alternatively, sterile vented plastic spike can be used to connect the68Ge / 68Ga generator’s outlet line in the next step.

[0398] e. Connect the male luer of the generator’s outlet line to longer sterile elution needle (e.g., 1”-23 / 4”) or use a sterile vented plastic spike.

[0399] f. Insert the sterile elution needle (or sterile vented plastic spike) into the vial through the rubber stopper.

[0400] g. Elute the generator directly into the vial according to the instructions for use of the GalliaPharm generator to get a net volume of 5 mL of eluate into the vial. Perform the elution manually or by means of a pump.

[0401] h. Disconnect the vial from the generator by removing the elution needle and vent needle with the 0.2 μm vent filter from the rubber stopper and swirl and invert the vial for 15-30 seconds to dissolve its contents.

[0402] i. Let the vial stand upright for 5 minutes at room temperature.LGPM Ref.: 773621: IST-001PC

[0403] j. After 5 minutes, assay the vial containing the gallium (68Ga) gozetotide for total radioactivity using a dose calibrator, calculate the radioactivity concentration and record the result.

[0404] k. The solution is ready to use after successful quality control.

[0405] l. Prior to use, visually inspect the solution behind a shielded screen for radioprotection purposes. Only use solutions that are clear without visible particles. m. Store the vial containing the gallium (68Ga) gozetotide solution upright in a lead shield below 25 °C until use. At the time of administration, the product must be aseptically withdrawn, and the radioprotection standards must be followed. The patient dose should be measured by a suitable radioactivity calibration system immediately prior to patient administration. Product administration data should also be recorded.

[0406] n. After reconstitution and radiolabeling and successful quality control, gallium (68Ga) gozetotide solution for injection can be diluted with water for injections or sodium chloride 9 mg / mL (0.9%) solution for injection up to a final volume of 10 mL.

[0407] For reconstitution and radiolabeling with IRE ELiT GalliAd / GalliEo Generator: a. Before use, bring the product vial to room temperature for a minimum of 10 minutes.

[0408] b. Remove the cap from the top of the vial and sanitize the closure of the vial with an appropriate alcohol pad to disinfect the surface and allow to air dry.

[0409] c. Place the vial in a suitable shielding.

[0410] d. Using a syringe fitted with a very fine needle (max. 29G, e.g. insulin syringe. (A fixed needle is recommended). Inject 0.5-1 mL of sterile water for injections into the vial through the surrounding of the stopper. The use of a very fine needle is essential to maintain the negative pressure inside the vial.

[0411] e. Use a new container of water for injections to prevent introduction of metal impurities

[0412] f. Remove the syringe and reconstitute the vial’s contents.

[0413] g. Connect the male luer of the generator’s outlet line to a non-metallic or coated sterile elution needle (e.g., 25G).

[0414] h. Turn the generator button to the loading position and wait for at least 10 seconds. Then, turn back the button to its initial position.

[0415] I. Insert the elution needle into the vial through the rubber stopper.

[0416] j. Elute the generator directly into the vial according to the instructions for use of the GalliAd generator to get a net volume of 1.1 mL of eluate into the reconstituted vial.LGPM Ref.: 773621: IST-001PC

[0417] k. Disconnect the vial from the generator by removing the elution needle. Swirl and invert the vial for 15-30 seconds to dissolve its contents.

[0418] l. Let the vial stand upright for at least 5 minutes at room temperature. m. After 5 minutes, assay the vial containing the gallium (68Ga) gozetotide for total radioactivity using a dose calibrator, calculate the radioactivity concentration and record the result.

[0419] n. The solution is ready to use after successful quality control.

[0420] o. Prior to use, visually inspect the solution behind a shielded screen for radioprotection purposes. Only use solutions that are clear without visible particles. p. Store the vial containing the gallium (68Ga) gozetotide solution upright in a lead shield below 25 °C until use. At the time of administration, the product must be aseptically withdrawn, and the radioprotection standards must be followed. The patient dose should be measured by a suitable radioactivity calibration system immediately prior to patient administration. Product administration data should also be recorded.

[0421] q. After reconstitution and radiolabeling and successful quality control, gallium (68Ga) gozetotide solution for injection can be diluted with water for injections or sodium chloride 9 mg / mL (0.9%) solution for injection up to a final volume of 10 mL.

[0422] Gallium (68Ga) gozetotide solution is stable up to 4 hours after preparation.

[0423] Therefore, the radiolabeled solution can be used within 4 hours after preparation according to the radioactivity required for the administration.

[0424] The disclosed subject matter is not to be limited in scope by the specific embodiments and examples described herein. Indeed, various modifications of the disclosure in addition to those described will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0425] All references (e.g., publications or patents or patent applications) cited herein are incorporated herein by reference in their entirety and for all purposes to the same extent as if each individual reference (e.g., publication or patent or patent application) was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Other embodiments are within the following claims.

Claims

LGPM Ref.: 773621: IST-001PCCLAIMS1. A radiolysis stabilized pharmaceutical composition comprising a radiopharmaceutical agent and gelatin.

2. The pharmaceutical composition of claim 1, wherein the radiopharmaceutical agent is a radiotherapeutic agent or a radioimaging agent.

3. The pharmaceutical composition of claim 1 or 2, wherein the radiopharmaceutical agent is a PSMA binding ligand chelated or bound to a radioisotope.

4. The pharmaceutical composition of claim 3, wherein the PSMA binding ligand chelated or bound to a radioisotope is selected from the group consisting of PSMA l& T, MCG, DCIT, (F-BA)KuE, (l-BA)KuE, DOTA-FFK(Sub-KuE), HBED-CC-Ahx, KuE, MIP-1404, MIP-1405, MIP-1072, MIP-1375, MIP-1095, MIP-1466, DUPA, PSMA5, PSMA6, rhPSMA, THP-PSMA, PSMA-11, PSMA-617, SFB, CTT-1298, CTT-1057, DCFBC, DCFPyL, YC-88, PSMA-1007, rhPSMA-7, and FSU-880.

5. The pharmaceutical composition of claim 3 or 4, wherein the PSMA binding ligand chelated to a radioisotope is selected from the group consisting of PSMA l& T, PSMA-11, and PSMA-1007.

6. The pharmaceutical composition of any one of claims 3-5, wherein the PSMA binding ligand chelated or bound to a radioisotope is PSMA l& T having the following structure:

7. The pharmaceutical composition of any one of claims 3-5, wherein the PSMA binding ligand chelated or bound to a radioisotope is PSMA-11 having the following structure:LGPM Ref.: 773621: IST-001PCHNHO2C ''-“CO2H8. The pharmaceutical composition of any one of claims 3-7, wherein the radioisotope is an alpha-emitting isotope.

9. The pharmaceutical composition of any one of claims 3-7, wherein the radioisotope is a beta-emitting isotope,10. The pharmaceutical composition of any one of claims 3-7, wherein the radioisotope is a gamma-emitting isotope.

11. The pharmaceutical composition of any one of claims 3-7, wherein the radioisotope is a positron-emitting isotope.

12. The pharmaceutical composition of any one of claims 3-7, wherein the radioisotope is selected from the group consisting of225Ac,111Ag,211At.198Au,199Au,212Bi,213Bi,11C,55Co,57Co,64Cu,67Cu,165Dy,166Dy,18F,67Ga,88Ga,152Gd,153Gd,157Gd,157Gd,166Ho,123l,124I,131I,111In,192Ir,140La,177Lu,13N,15O,212Pb,103Pd,109Pd,149Pm,223Ra,224Ra,186Re,188Re,105Rh,43Sc,44Sc,47Sc,153Sm,149Tb,151Tb,161Tb,94Tc,99mTc,227Th,86Y,90Y,169Yb,175Yb, and89Zr.

13. The pharmaceutical composition of any one of claims 3-7 and 12, wherein the radioisotope is selected from the group consisting of161Tb,177Lu,18F, and68Ga.

14. The pharmaceutical composition of any one of claims 3-7, 12, and 13, wherein the radioisotope is161Tb.

15. The pharmaceutical composition of any one of claims 3-7, 12, and 13, wherein the radioisotope is177Lu.LGPM Ref.: 773621: IST-001PC16. The pharmaceutical composition of any one of claims 3-7, 12, and 13, wherein the radioisotope is68Ga.

17. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises161Dy.

18. The pharmaceutical composition of any one of claims 1-17, wherein the gelatin is selected from the group consisting of type A gelatin, type B gelatin, type AB gelatin, and hydrolyzed gelatin.

19. The pharmaceutical composition of any one of claims 1-18, wherein the gelatin is hydrolyzed gelatin.

20. The pharmaceutical composition of any one of claims 1-19, wherein the gelatin is a non-gelling gelatin.

21. The pharmaceutical composition of any one of claims 1-20, wherein the pharmaceutical composition further comprises a buffer.

22. The pharmaceutical composition of claim 21, wherein the buffer is selected from the group consisting of ascorbate, acetate, carbonate, citrate, histidine, phosphate, tartrate, and Tris buffers.

23. The pharmaceutical composition of claim 21 or 22, wherein the buffer is an ascorbate buffer.

24. The pharmaceutical composition of claim 21 or 22, wherein the buffer is an acetate buffer.

25. The pharmaceutical composition of any one of claims 1-24, wherein the pH of the pharmaceutical composition at the end of production of the pharmaceutical composition is about 4 to about 5.

26. The pharmaceutical composition of any one of claims 1-25, wherein the pharmaceutical composition further comprises saline.LGPM Ref.: 773621: IST-001PC27. The pharmaceutical composition of any one of claims 1-26, wherein the pharmaceutical composition has at least about 99% radiochemical purity (RCP) by HPLC at the end of production of the pharmaceutical composition.

28. The pharmaceutical composition of any one of claims 1-27, wherein the pharmaceutical composition has at least about 98% RCP by HPLC about 45 hours after production of the pharmaceutical composition.

29. The pharmaceutical composition of any one of claims 1-28, wherein the pharmaceutical composition has at least about 97% RCP by HPLC about 70 hours after production of the pharmaceutical composition.

30. The pharmaceutical composition of any one of claims 1-29, wherein the pharmaceutical composition has at least about 94% RCP by HPLC about 140 hours after production of the pharmaceutical composition.

31. The pharmaceutical composition of any one of claims 1-6, 12-14, 18-23, and 25-30, wherein the pharmaceutical composition comprises:a) PSMA l& T radiolabeled with161Tb;b) hydrolyzed gelatin;c) an ascorbate buffer; andd) saline.

32. The pharmaceutical composition of any one of claims 1-6, 12-14, and 18-31, wherein the pharmaceutical composition has a ratio of PSMA l& T (pg) to161Tb (mCi) of about 1:2 to about 1:1.

33. The pharmaceutical composition of any one of claims 1-6, 12, 13, 15, 18-23, and 25-30, wherein the pharmaceutical composition comprises:a) PSMA l& T radiolabeled with177Lu;b) hydrolyzed gelatin;c) an ascorbate buffer; andd) saline.

34. The pharmaceutical composition of any one of claims 1-5, 7, 12, 13, 16, 18-22, and 24- 30, wherein the pharmaceutical composition comprises:a) PSMA-11 radiolabeled with68Ga;LGPM Ref.: 773621: IST-001PCb) hydrolyzed gelatin;c) an acetate buffer; andd) saline.

35. The pharmaceutical composition of any one of claims 1, 17-23, and 25-30, wherein the pharmaceutical composition comprises:a) PSMA l& T labeled with161Dy;b) gelatin;c) an ascorbate buffer; andd) saline.

36. The pharmaceutical composition of any one of claims 1-35, wherein the pharmaceutical composition comprises about 20 to about 100 mg of gelatin.

37. The pharmaceutical composition of any one of claims 1-36, wherein the pharmaceutical composition comprises about 30 to about 70 mg of gelatin.

38. The pharmaceutical composition of any one of claims 1-37, wherein the pharmaceutical composition comprises about 30 to about 50 mg of gelatin.

39. The pharmaceutical composition of any one of claims 1-38, wherein the pharmaceutical composition comprises about 35 mg of gelatin.

40. A method of treating cancer in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition of any one of claims 1-39.

41. The method of claim 40, wherein the cancer is associated with PSMA activity.

42. The method of claim 40 or 41, wherein the cancer is prostate cancer.

43. The method of any one of claims 40-42, wherein the cancer is metastatic.

44. A method of diagnosing cancer in a subject comprising administering to the subject a pharmaceutical composition of any one of claims 1-39.LGPM Ref.: 773621: IST-001PC45. Use of the pharmaceutical composition of any one of claims 1-39 as an imaging agent.

46. The use of claim 45, wherein the imaging agent is used in positron emission tomography.

47. The use of claim 45 or 46, wherein the imaging agent is used to visualize cell death.

48. A method of diagnosing or treating a disease or disorder in a subject in need thereof comprising administering to the subject an effective amount of a pharmaceutical composition of any one of claims 1-39.

49. The method of claim 48, wherein the disease or disorder is cancer.

50. The method of claim 48 or 49, wherein the disease or disorder is cancer associated with PSMA activity.

51. The method of claim any one of claims 48-50, wherein the disease or disorder is prostate cancer.

52. The method of any one of claims 49-51, wherein the cancer is metastatic.

53. The method of any one of claims 48-52 further comprising conducting positron emission tomography on the subject following administration of the pharmaceutical composition.

54. The method or use of any one of claims 40-53, wherein the pharmaceutical composition is administered intravenously.

55. The method or use of any one of claims 40-54, wherein the pharmaceutical composition is administered by infusion.

56. The method or use of any one of claims 40-55, wherein the pH of the pharmaceutical composition at the time of administrating the pharmaceutical composition is about 4 to about 8.LGPM Ref.: 773621: IST-001PC57. The method or use of any one of claims 40-56, wherein the pH of the pharmaceutical composition at the time of administrating the pharmaceutical composition is about 7.

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