Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
  • C07K7/02—Linear peptides containing at least one abnormal peptide link
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
  • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
  • A61K51/04—Organic compounds
  • A61K51/0404—Lipids, e.g. triglycerides; Polycationic carriers
  • A61K51/0406—Amines, polyamines, e.g. spermine, spermidine, amino acids, (bis)guanidines
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
  • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
  • A61K51/04—Organic compounds
  • A61K51/0402—Organic compounds carboxylic acid carriers, fatty acids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
  • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
  • A61K51/04—Organic compounds
  • A61K51/0497—Organic compounds conjugates with a carrier being an organic compounds
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
  • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
  • C07B59/001—Acyclic or carbocyclic compounds
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D257/00—Heterocyclic compounds containing rings having four nitrogen atoms as the only ring hetero atoms
  • C07D257/02—Heterocyclic compounds containing rings having four nitrogen atoms as the only ring hetero atoms not condensed with other rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D273/00—Heterocyclic compounds containing rings having nitrogen and oxygen atoms as the only ring hetero atoms, not provided for by groups C07D261/00 - C07D271/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2121/00—Preparations for use in therapy
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
  • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
  • C07B2200/05—Isotopically modified compounds, e.g. labelled

Definitions

• the present invention relates to radiolabelled compounds for treatment of diseases or conditions characterized by expression of prostate-specific membrane antigen, particularly compounds with low uptake in salivary glands and/or kidneys.
• PSMA Prostate-specific membrane antigen
• PSMA is a transmembrane protein that catalyzes the hydrolysis of /V-acetyl-aspartylglutamate to glutamate and /V-acetylaspartate.
• PSMA is selectively overexpressed in certain diseases and conditions compared to most normal tissues. For example, PSMA is overexpressed up to 1 ,000-fold in prostate tumors and metastases. Due to its pathological expression pattern, various radiolabeled PSMA-targeting constructs have been designed and evaluated for imaging of PSMA-expressing tissues and/or for therapy of diseases or conditions characterized by PSMA expression.
• a number of radiolabeled PSMA-targeting derivatives of lysine-urea-glutamate have been developed, including 18 F-DCFBC, 18 F-DCFPyL, 68 Ga-PSMA-HBED-CC, 68 Ga- PSMA-617, 68 Ga-PSMA I & T (see Figure 1) as well as versions of the foregoing labelled with alpha emitters (such as 225 Ac) or beta emitters (such as 177 Lu or 90 Y).
• PSMA-617 radiolabeled with therapeutic radionuclides such as 177 Lu and 225 Ac
• therapeutic radionuclides such as 177 Lu and 225 Ac
• mCRPC metastatic castration resistant prostate cancer
• dry mouth (xerostomia) altered taste and adverse renal events are common side effects of this treatment, due to high salivary gland and kidney accumulation of the radiotracer
• Radiotracer accumulation in the kidneys and salivary gland is therefore a limiting factor that reduces the maximal cumulative administered activity that can be safely given to patients, which limits the potential therapeutic effectiveness of Lys-urea-Glu based radiopharmaceuticals (Violet et al. 2019 J Nucl Med. 60(4):517-523).
• This disclosure relates to PSMA-targeting compounds that bind PSMA using a Lys-ureido- Glu moiety or derivative moiety thereof, a radiometal chelator, and an albumin binder.
• the positions of both the radiometal chelator and the albumin binder relative to the PSMA-binding moiety result in novel compounds with useful delivery of radiation to tumor site(s) and improved side effects (e.g. low uptake in kidneys and/or salivary gland).
• the disclosed compounds may minimize structural hinderance.
• FIGURE 1 shows examples of prior art PSMA-targeting compounds for prostate cancer imaging.
• FIGURE 2A shows representative in vitro competitive PSMA binding assay curves using LNCaP cells with CCZ02009 (left panel) and HTK03170 (right panel).
• Figure 2B shows PET images of 68 Ga-CCZ02009 in NRG-mice bearing LNCaP tumors at 1 (left panel) and 3 h p.i. (right panel).
• FIGURE 3 shows representative in vitro competitive PSMA binding assay curves using LNCaP cells with CCZ02060 (left panel) and CCZ02059 (right panel).
• FIGURE 4 shows SPECT/CT images of 177 Lu-CCZ02017 in NRG-mice bearing LNCaP tumors at 3, 24, 72, 144 and 240 h p.i.
• FIGURE 5 shows a representative in vitro competitive PSMA binding assay curve using LNCaP cells with CCZ02008.
• FIGURE 6 shows a representative in vitro competitive PSMA binding assay curve using LNCaP cells with CCZ02025.
• FIGURE 7 shows a representative in vitro competitive PSMA binding assay curve using LNCaP cells with CCZ02024.
• FIGURE 8 shows a representative in vitro competitive PSMA binding assay curve using LNCaP cells with CCZ02015.
• FIGURES 9A-9B show representative in vitro competitive PSMA binding assay curves using LNCaP cells with CCZ02012 (FIGURE 9A) and CCZ02013 (FIGURE 9B).
• FIGURES 10A-10B show representative in vitro competitive PSMA binding assay curves using LNCaP cells with CCZ02021 (FIGURE 10A) and CCZ02022 (FIGURE 10B).
• FIGURE 11 shows a representative in vitro competitive PSMA binding assay curve using LNCaP cells with CCZ02034.
• FIGURE 12 shows a representative in vitro competitive PSMA binding assay curve using LNCaP cells with CCZ02005.
• FIGURE 13 shows a representative in vitro competitive PSMA binding assay curve using LNCaP cells with CCZ02061 .
• the terms “comprising,” “having”, “including” and “containing,” and grammatical variations thereof, are inclusive or open-ended and do not exclude additional, unrecited elements and/or method steps, even if a feature/component defined as a part thereof consists or consists essentially of specified feature(s)/component(s).
• compositions, use or method excludes the presence of additional elements and/or method steps in that feature.
• a compound, composition, use or method described herein as comprising certain elements and/or steps may also, in certain embodiments consist essentially of those elements and/or steps, and in other embodiments consist of those elements and/or steps, whether or not these embodiments are specifically referred to.
• a use or method described herein as comprising certain elements and/or steps may also, in certain embodiments consist essentially of those elements and/or steps, and in other embodiments consist of those elements and/or steps, whether or not these embodiments are specifically referred to.
• a reference to an element by the indefinite article “a” does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there be one and only one of the elements.
• the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise.
• the use of the word “a” or “an” when used herein in conjunction with the term “comprising” may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one” and “one or more than one.”
• the terms “treat”, “treatment”, “therapeutic” and the like includes ameliorating symptoms, reducing disease progression, improving prognosis and reducing recurrence.
• the term “subject” refers to an animal (e.g. a mammal or a non-mammal animal).
• the subject may be a human or a non-human primate.
• the subject may be a laboratory mammal (e.g., mouse, rat, rabbit, hamster and the like).
• the subject may be an agricultural animal (e.g., equine, ovine, bovine, porcine, camelid and the like) or a domestic animal (e.g. , canine, feline and the like).
• the subject is a human.
• the compounds disclosed herein may also include base-free forms, salts or pharmaceutically acceptable salts thereof. Unless otherwise specified, the compounds claimed and described herein are meant to include all racemic mixtures and all individual enantiomers or combinations thereof, whether or not they are explicitly represented herein.
• the compounds disclosed herein may be shown as having one or more charged groups, may be shown with ionizable groups in an uncharged (e.g. protonated) state or may be shown without specifying formal charges.
• the ionization state of certain groups within a compound e.g. without limitation, CO 2 H, PO 3 H 2 , SO 2 H, SO3H, SO4H, OPOSH 2 and the like
• a carboxylic acid group i.e.
• COOH would be understood to usually be deprotonated (and negatively charged) at neutral pH and at most physiological pH values, unless the protonated state is stabilized.
• OSO 3 H i.e. SO 4 H
• SO 2 H groups SO 3 H groups
• OPO 3 H 2 i.e. PO 4 H 2
• PO 3 H groups would generally be deprotonated (and negatively charged) at neutral and physiological pH values.
• salts and solvate have their usual meaning in chemistry.
• the compound when it is a salt or solvate, it is associated with a suitable counter-ion.
• a suitable counter-ion It is well known in the art how to prepare salts or to exchange counter-ions.
• such salts can be prepared by reacting free acid forms of these compounds with a stoichiometric amount of a suitable base (e.g. without limitation, Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, orthe like), or by reacting free base forms of these compounds with a stoichiometric amount of a suitable acid. Such reactions are generally carried out in water or in an organic solvent, or in a mixture of the two.
• Counter-ions may be changed, for example, by ion-exchange techniques such as ion-exchange chromatography. All zwitterions, salts, solvates and counter-ions are intended, unless a particular form is specifically indicated.
• the salt or counter-ion may be pharmaceutically acceptable, for administration to a subject.
• suitable excipients include any suitable buffers, stabilizing agents, salts, antioxidants, complexing agents, tonicity agents, cryoprotectants, lyoprotectants, suspending agents, emulsifying agents, antimicrobial agents, preservatives, chelating agents, binding agents, surfactants, wetting agents, non-aqueous vehicles such as fixed oils, or polymers for sustained or controlled release. See, for example, Berge et al. 1977. (J. Pharm Sci. 66:1-19), or Remington- The Science and Practice of Pharmacy, 21st edition (Gennaro et al editors. Lippincott Williams & Wilkins Philadelphia) , each of which is incorporated by reference in its entirety.
• C n where n is an integer (e.g. 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16,17, 18, 19, 20, and the like) or where n is defined as a range of integers (e.g. 1-20, 1-18, 2-15, 3-20, and the like) refers to the number of carbons in a compound, R-group, L- group, or substituent, or refers to the number of carbons plus heteroatoms in a compound, R-group, L-group, or substituent.
• a range of integers includes all integers in the range; e.g.
• heteroatoms may include any, some or all possible heteroatoms.
• the heteroatoms may be selected from N, O, S, P and Se.
• the heteroatoms are selected from N, S, or O.
• Such embodiments are non-limiting.
• alkyl has at least one carbon-carbon double bond, and may have any number of carbon-carbon single bonds.
• alkynylenyl has at least one carbon-carbon triple bond, and may have any number of carbon-carbon single bonds.
• alkylenyl, alkenylenyl and/or alkynylenyl and “alkylenyl, alkenylenyl or alkynylenyl” are intended to be equivalent and each includes hydrocarbon chains that can have any reasonable number or combination of carbon-carbon single bonds, double bonds, and triple bonds. These hydrocarbon chains can be linear, branched, cyclic, or any combination of linear and branched, linear and cyclic, cyclic and branched, branched and cyclic, or linear, branched and cyclic. Cyclic hydrocarbons may be nonaromatic, partially aromatic, or aromatic. Unless otherwise specified, the term “cyclic” includes single rings, multiple non-fused rings, fused rings, bridged rings, and combinations thereof.
• any carbon ... is optionally independently replaced by N, S, or O
• the defined hydrocarbon e.g. “alkyl”, “alkylenyl”, “alkenylenyl”, or “alkynylenyl”
• the defined hydrocarbon includes zero, one, more than one, or any reasonable combination of two or more heteroatoms selected from N, S, and O.
• the above expression therefore expands the defined hydrocarbon to additionally encompass heteroalkyls, heteroalkylenyls, heteroalkenylenyls, and heteroalkynylenyls, etc.
• the person of skill in the art would understand that various combinations of different heteroatoms may be used.
• any carbon bonded to two other carbons is optionally independently replaced by N, S, or O
• any carbon in the defined hydrocarbon bonded to two other carbons e.g. the underlined carbon in -C-C-C-
• whetherthose bonds are single, double, ortriple bonds may be a heteroatom, but excludes heteroatoms bonded to other heteroatoms (e.g. excludes -C-N-S-, -S- S-N-, -N-S-C-, and the like).
• R-groups e.g. R 1 , R 2 , R 3 , etc.
• L-groups e.g. L 1 , L 2 , L 3 , etc.
• L-groups g N(alkyl) If unspecified, the size of an R-group or L-group is what would be considered reasonable to the person of skill in the art.
• the size of an alkyl may be 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 89, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, 100 or more than 100 carbons in length, subject to the common general knowledge of the person of skill in
• the size of a heteroalkyl may be 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35,
• alkyl, alkenyl or alkynyl and similar expressions, the “alkyl” would be understood to be a saturated alkyl, and the “alkenyl” and the “alkynyl” would be understood to be unsaturated.
• linear may be used as it is normally understood to a person of skill in the art and generally refers to a chemical entity that comprises a skeleton or main chain that does not split off into more than one contiguous chain.
• linear alkyls include methyl, ethyl, n-propyl, and n-butyl.
• branched may be used as it is normally understood to a person of skill in the art and generally refers to a chemical entity that comprises a skeleton or main chain that splits off into more than one contiguous chain.
• the portions of the skeleton or main chain that split off in more than one direction may be linear, cyclic or any combination thereof.
• Non-limiting examples of a branched alkyl group include tert-butyl and isopropyl.
• alkylenyl refers to a divalent analog of an alkyl group.
• alkylenyl, alkenylenyl and/or alkynylenyl the “alkylenyl” would be understood to be a saturated alkylenyl, and the “alkenylenyl” and the “alkynylenyl” would be understood to be unsaturated.
• heteroalkylenyl refers to a divalent analog of a heteroalkyl group.
• heteroalkenylenyl refers to a divalent analog of a heteroalkenyl group.
• heteroalkynylenyl refers to a divalent analog of a heteroalkynyl group.
• saturated when referring to a chemical entity may be used as it is normally understood to a person of skill in the art and generally refers to a chemical entity that comprises only single bonds, and may include linear, branched, and/or cyclic groups.
• Non-limiting examples of a saturated C1-C20 alkyl group may include methyl, ethyl, n-propyl, i-propyl, sec-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, i-pentyl, sec-pentyl, t-pentyl, n-hexyl, i-hexyl, 1 ,2- dimethylpropyl, 2-ethylpropyl, 1-methyl-2-ethylpropyl, l-ethyl-2-methylpropyl, 1 , 1 ,2-trimethylpropyl, 1 ,1 ,2-triethylpropyl, 1 ,1 -dimethylbutyl, 2,2-dimethylbutyl, 2-ethylbutyl, 1 ,3-dimethylbutyl, 2- methylpentyl, 3-methylpentyl,
• the term “unsaturated” when referring to a chemical entity may be used as it is normally understood to a person of skill in the art and generally refers to a chemical entity that comprises at least one double or triple bond, and may include linear, branched, and/or cyclic groups.
• Non-limiting examples of a C2-C20 alkenyl group may include vinyl, allyl, isopropenyl, I- propene-2-yl, 1-butene-l-yl, l-butene-2-yl, l-butene-3-yl, 2-butene-l-yl, 2-butene-2-yl, octenyl, decenyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclononanenyl, cyclodecanenyl, and the like.
• a C1-C20 alkenylenyl therefore encompasses, without limitation, all divalent analogs of the above-listed alkenyl groups.
• Non-limiting examples of a C2-C20 alkynyl group may include ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, and the like.
• a C1-C20 alkynylenyl therefore encompasses, without limitation, all divalent analogs of the above-listed alkynyl groups.
• Non-limiting examples of non-aromatic cyclic groups include cylcopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.
• Non-limiting examples of non-aromatic heterocyclic groups include aziridinyl, azetidinyl, diazetidinyl, pyrrolidinyl, pyrrolinyl, piperidinyl, piperazinyl, imidazolinyl, pyrazolidinyl, imidazolydinyl, phthalimidyl, succinimidyl, oxiranyl, tetrahydropyranyl, oxetanyl, dioxanyl, thietanyl, thiepinyl, morpholinyl, oxathiolanyl, and the like.
• an “aryl” group includes both single aromatic rings as well as fused rings containing at least one aromatic ring, non-limiting examples of C3-C20 aryl groups include phenyl (Ph), pentalenyl, indenyl, naphthyl and azulenyl.
• Non-limiting examples of aromatic heterocyclic groups of similar size include pyrrolyl, imidazolyl, pyrazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pirazinyl, quinolinyl, isoquinolinyl, acridinyl, indolyl, isoindolyl, indolizinyl, purinyl, carbazolyl, indazolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl, phenanthridinyl, phenazinyl, phenanthrolinyl, perimidinyl, furyl, dibenzofuryl, xanthenyl, benzofuryl, thiophenyl, thianthrenyl, benzothiophenyl, phosphorinyl, phosphinolin
• substituted is used as it would normally be understood to a person of skill in the art and generally refers to a compound or chemical entity that has one chemical group replaced with a different chemical group.
• a substituted alkyl, alkylenyl, alkenylenyl, or alkynylenyl has one or more hydrogen atom(s) independently replaced with an atom that is not hydrogen.
• chloromethyl is a non-limiting example of a substituted alkyl, more particularly an example of a substituted methyl.
• Aminoethyl is another non-limiting example of a substituted alkyl, more particularly an example of a substituted ethyl.
• a substituted compound or group may be substituted with any chemical group reasonable to the person of skill in the art.
• a hydrogen bonded to a carbon or heteroatom e.g. N
• halide e.g.
• each carbon may be independently substituted or unsubstituted with oxo, hydroxyl, sulfhydryl, amine, amide, urea, halogen, guanidino, carboxylic acid, sulfonic acid, sulfinic acid, or phosphoric acid.
• the amide substituent is -C(O)-NH 2 .
• unsubstituted is used as it would normally be understood to a person of skill in the art.
• Non-limiting examples of unsubstituted alkyls include methyl, ethyl, tertbutyl, pentyl and the like.
• the expression “optionally substituted” is used interchangeably with the expression “unsubstituted or substituted”.
• the expression “optionally independently substituted” means that each location may be substituted or may not be substituted, and when substituted each substituent may be the same or different.
• hydrogen may or may not be shown.
• hydrogens may be protium (i.e. 1 H), deuterium (i.e. 2 H) or combinations of 1 H and 2 H.
• Methods for exchanging 1 H with 2 H are well known in the art.
• solvent-exchangeable hydrogens the exchange of 1 H with 2 H occurs readily in the presence of a suitable deuterium source, without any catalyst.
• acid, base or metal catalysts coupled with conditions of increased temperature and pressure, can facilitate the exchange of nonexchangeable hydrogen atoms, generally resulting in the exchange of all 1 H to 2 H in a molecule.
• the compounds disclosed herein may be synthesized (at least in part) using peptide synthesis methods.
• Each amino acid residue in a peptide or peptidic region has both an amino group and a carboxylic acid group, either or both of which can be used for covalent attachment.
• the amino group and/or the carboxylic acid group may be converted to an amide or other structure; e.g. a carboxylic acid group of a first amino acid is converted to an amide (i.e. a peptide bond) when bonded to the amino group of a second amino acid.
• amino acid residues may have the formula -N(R a )-R b -C(O)-, where R a and R b are R-groups.
• R a will typically be hydrogen or methyl (or a different alkyl).
• the amino acid residues of a peptide may comprise typical peptide (amide) bonds and may further comprise bonds between side chain functional groups and the side chain or main chain functional group of another amino acid.
• the side chain carboxylate of one amino acid residue (e.g. Asp, Glu, etc.) in the peptide or peptidic region may be bonded to the amine of another amino acid residue (e.g. Dap, Dab, Orn, Lys) in the peptide or peptidic region.
• an amino acid may be any amino acid, including proteinogenic and nonproteinogenic amino acids, alpha amino acids, beta amino acids, or any other amino acid.
• nonproteinogenic amino acids are shown in Table 1 and include: D-amino acids (including without limitation any D-form of the following amino acids), ornithine (Orn), 3-(1- naphtyl)alanine (Nal), 3-(2-naphtyl)alanine (2-Nal), a-aminobutyric acid, norvaline, norleucine (Nle), homonorleucine, beta-(1 ,2,3-triazol-4-yl)-L-alanine, 1 ,2,4-triazole-3-alanine, Phe(4-F), Phe(4-CI), Phe(4-Br), Phe(4-I), Phe(4-NH 2 ), Phe(4-NO 2 ), homoarginine (hArg), 2-amino
• the wavy line ” symbol shown through or at the end of a bond in a chemical formula is intended to define the group on one side of the wavy line, without modifying the definition of the structure on the opposite side of the wavy line.
• any atoms shown outside the wavy lines are intended to clarify orientation of the defined group. As such, only the atoms between the two wavy lines constitute the definition of the R-group or L-group.
• atoms are not shown outside the wavy lines (e.g.
• each R 4 is independently a linear, branched, and/or cyclic C n 6 alkylenyl,
• R 5a -X br (R 5c )-R 5b forms a linear, branched, and/or cyclic C n ? alkylenyl, alkenylenyl and/or alkynylenyl, wherein n7 is 1-20, wherein any carbon bonded to two other carbons is optionally independently replaced by N, S, or O, wherein carbons are optionally independently substituted with oxo, hydroxyl, sulfhydryl, amine, amide, urea, halogen, guanidino, carboxylic acid, sulfonic acid, sulfinic acid, or phosphoric acid, wherein one, two, or three of R 5a , R 5b , and R 5c are optionally absent, wherein X br is C, CH, or N and wherein X br is separated from ring A by at least 4 atoms; each R 6 and each R 7 is independently a linear, branched, and/or cyclic C
• R rad is a radiometal chelator, optionally bound by a radiometal, wherein R rad is separated from ring A by at least 7 atoms;
• R alb is an albumin binder, wherein the albumin binder is:
• n9 is 8-20;
• n10 is 8-20; or wherein n11 is 1-4 and R 8 is I, Br, F, Cl, H, OH, OCH 3 , NH 2 , NO 2 , or CH 3 ; and
• R alb is separated from ring A by at least 7 atoms.
• the compound (of Formula I) has Formula II or is a salt or a solvate of Formula II: defined in Formula I, or as defined in any other embodiment(s) defined herein.
• R 1 is-CH 2 -CH 2 - or-CHF-.
• R 2 is-(CH 2 ) 4 -
• L 1 is -NH-C(O)-.
• L 2 is -NHC(O)-.
• n1 is 0; ring A has 0 double bonds and is bonded at para position, optionally wherein ring some of these embodiments, n2 is 0 or 1.
• each R L3a is H.
• X br is N or CH.
• R 5b is -(CH 2 ) 4 - and R 5c is absent; or R 5c is -(CH 2 ) 4 - and R 5b is absent.
• L 5b is -NH-C(O)-.
• L 5c is -NH-C(O)-.
• L 5b is is -NH-C(O)-.
• n4 is 0 or 1
• R 6 — when present — is methylene.
• n5 is 0 or 1
• R 7 — when present — is methylene.
• L 6 — when present — is -NH-C(O)-.
• L 7 — when present — is -NH-C(O)-.
• R alb is
• R rad is DOTA or a DOTA derivative.
• R 1 is -R 1a R 1b -, wherein R 1a is absent or -CH 2 -, and R 1b is-CH 2 -or -CHF;
• R 2 is -(CH 2 ) 3 -O- , -(CH 2 ) 3 -, -(CH 2 ) 4 -, or -CH 2 -0-(CH 2 ) 2 -;
• L 1 is -N(R L1a )-C(O)-, -C(O)-N(R L1a )-, or -NH-C(O)-NH-, wherein R L1a is H or methyl;
• L 2 is -N(R L2a )-C(O)-, -C(O)-N(R L2a )-, -NH-C(O)-NH- , wherein R L2a is H, or methyl; each of n1 and n2 is 0-2; ring A has 0 bonds, and is bonded at meta or para position; each of L 3 , L 4 , L 5b , L 5c , L 6 , and L 7 is independently -N(R L3a )-C(O)-, -C(O)-N(R L3a )-, or -NH- C(O)-NH-, wherein each R L3a is independently H, methyl, or ethyl; each of n3, n4, and n5 is independently 0-4; each R 4 is independently a linear C1-C10 alkylenyl;
• R 5a -X br (R 5c )-R 5b forms a linear, branched, and/or cyclic Ci.C 20 alkylenyl, alkenylenyl and/or alkynylenyl, wherein any carbon bonded to two other carbons is optionally independently replaced by N, S, or O, wherein carbons are optionally independently substituted with oxo, hydroxyl, sulfhydryl, amine, amide, urea, halogen, guanidino, carboxylic acid, sulfonic acid, sulfinic acid, or phosphoric acid, wherein one, two, or three of R 5a , R 5b , and R 5c are optionally absent, wherein X br is N or CH, and wherein X br is separated from ring A by at least 4 atoms; each R 6 and each R 7 is independently a linear, branched, and/or cyclic C1-C20 alkylenyl, alken
• R rad is a radiometal chelator, optionally bound by a radiometal, wherein R rad is separated from ring A by at least 7 atoms;
• R alb is: wherein n11 is 1-4 and R 8 is I, Br, F, Cl, H, OH, OCH 3 , NH 2 ,
• R 2 is -(CH 2 )3-, or -(CH 2 ) 4 -;
• L 1 is -N(R L1a )-C(O)-, or -C(O)-N(R L1a )-, wherein R L1a is H or methyl;
• R 3 is:
• L 2 is -N(R L2a )-C(O)-, or -C(O)-N(R L2a )-, wherein R L2a is H, or methyl; each of n1 and n2 is 0-1 ; each of L 3 , L 4 , L 5b , L 5c , L 6 , and L 7 is independently -N(R L3a )-C(O)-, or -C(0)-N(R L3a )-, wherein each R L3a is independently H or methyl; each R 4 is independently a linear C1-C10 alkylenyl;
• R 5a -X br (R 5c )-R 5b forms a linear, branched, and/or cyclic C1-C10 alkylenyl, wherein any carbon bonded to two other carbons is optionally independently replaced by O, wherein carbons are optionally independently substituted with oxo, hydroxyl, amine, amide, or carboxylic acid, wherein one, two, or three of R 5a , R 5b , and R 5c are optionally absent, wherein X br is N or CH and wherein X br is separated from ring A by at least 4 atoms; each R 6 and each R 7 is independently a linear, branched, and/or cyclic C1-C10 alkylenyl, wherein any carbon bonded to two other carbons is optionally independently replaced by N, S, or O, and carbons are optionally independently substituted with oxo, hydroxyl, amine, amide, urea, carboxylic acid; R alb is : wherein n
• L 3 is -NHC(O)-.
• L 4 is - NHC(O)-.
• R 4 is methylene.
• n3 is 1-4.
• n3 is 1 -2.
• n3 is 1.
• n2 is 0-1.
• n2 is 1.
• R 5a is absent.
• R 5a is methylene.
• R 1a is absent. In some embodiments, R 1a is -CH2-. In some embodiments, R 1a is -O-. In some embodiments, R 1a is -S-
• R 1b is -CH 2 - In some embodiments, R 1b is -CHF-.
• R 1 is -CH 2 - In some embodiments, R 1 is -CHF-. In some embodiments, R 1 is -CH 2 -CH 2 - In some embodiments, R 1 is-CH 2 -CHF- In some embodiments, R 1 is -O-CH 2 - In some embodiments, R 1 is -O-CHF- In some embodiments, R 1 is -S-CH 2 - In some embodiments, R 1 is -S-CHF-.
• R 2 is -(CH 2 )3-O- In some embodiments, R 2 is -(CH 2 )3- In some embodiments, R 2 is -(CH 2 ) 4 - In some embodiments, R 2 is -CH 2 -O-(CH 2 ) 2 -. In some embodiments, R 2 is -CH 2 -S-(CH 2 ) 2 -.
• L 1 is -N(R L1a )-C(O)- or-C(0)-N(R L1a )- wherein R L1a is as defined in Formula I.
• R L1a is H.
• R L1a is methyl.
• R L1a is ethyl.
• R L1a is a benzyl group with 0-4 substituents independently selected from halogen, OMe, or SMe.
• R L1a is a benzyl group.
• L 1 is -N(R L1a )-C(O)- or -C(O)- N(R L1a )- wherein L 1a is H or methyl.
• L 1 is-NHC(O)-.
• L 1 is -C(O)-NH-.
• R 5a -X br (R 5c )-R 5b forms a linear or branched Ci.C 20 alkylenyl, wherein any carbon bonded to two other carbons is optionally independently replaced by O, wherein carbons are optionally independently substituted with oxo, hydroxyl, amine, amide, or carboxylic acid, wherein one, two, or three of R 5a , R 5b , and R 5c are optionally absent, wherein X br is N or CH and wherein X br is separated from ring A by at least 4 atoms.
• R 5a -X br (R 5c )-R 5b forms a linear or branched C1-C10 alkylenyl, wherein any carbon bonded to two other carbons is optionally independently replaced by O. In another specific embodiment, 1 , 2, 3 or 4 carbons are replaced by O.
• L 1 is -S-. In some embodiments, embodiments, NH-. In some embodiments, embodiments, some embodiments, L 1 is In some embodiments, L 1 , substituted with 0-4 substituents independently selected from C1-C4 alkyl, halogen, OMe, SMe, NH 2 , NO 2 , CN, or OH, and wherein
• R 3 is: substituted with 0-4 substituents independently selected from C1-C4 alkyl, halogen, OMe, SMe,
• R 3 0-4 ring carbons are replaced with nitrogen; in some such embodiments, the rings are unsubstituted and contain a single nitrogen.
• R 3 0-4 ring carbons are replaced with nitrogen; in some such embodiments, the rings are unsubstituted and contain a single nitrogen.
• L 2 is -N(R L2a )-C(O)- or-C(0)-N(R L2a )- wherein R L2a is as defined in Formula I.
• R L2a is H.
• R L2a is methyl.
• R L2a is ethyl.
• R L2a is a benzyl group with 0-4 substituents independently selected from halogen, OMe, or SMe.
• R L2a is a benzyl group.
• L 2 is -N(R L2a )-C(O)- or -C(O)- N(R L2a )- wherein L 2a is H or methyl.
• L 2 is-NHC(O)-.
• L 2 is -C(O)-NH-.
• L 2 is -S-. In some embodiments, some embodiments, NH-. In some embodiments, some embodiments, L 2 is In some embodiments, L 2
• n1 is 0. In some embodiments, n1 is 1 . In some embodiments n1 is 2.
• ring A has 0 double bonds (i.e. all single bonds). In some embodiments, ring A has 1 double bond. In some embodiments, ring A has 2 double bonds. In some embodiments, ring A has 3 double bonds. In some embodiments, ring A is bonded at meta position. In some embodiments, ring A is bonded at para position. In some embodiments, ring A has 0 double bonds and is bonded at para position. In some embodiments, ring A is some embodiments, ring A is some embodiments, ring A is
• n2 is 0. In some embodiments, n2 is 1 . In some embodiments n2 is 2.
• L 3 is -N(R L3a )-C(O)- or-C(0)-N(R L3a )- wherein R L3a is as defined in Formula I.
• R L3a is H.
• R L3a is methyl.
• R L3a is ethyl.
• L 3 is -N(R L3a )-C(O)- or -C(O)- N(R L3a )- wherein R L3a is H or methyl.
• L 3 is -NHC(O)-.
• L 3 is -C(O)-NH- In some embodiments, L 3 is -S-. In some embodiments, L 3 is - NH-C(O)-NH-. In some embodiments, L 3 is -NH-C(S)-NH-. In some embodiments, L 3 is , In some embodiments, L 3 is some embodiments,
• R 1 is -CH?- CH:- and R 2 is -(CH2)4-.
• L 1 some such embodiments. optionally wherein R 3 is some such embodiments, L 2 is -NHC(O)-.
• n1 is O; ring A has 0 double bonds and is bonded at para position, optionally wherein ring A is some such embodiments, n2 is 0 or 1 .
• L 3 is -
• n3 is 0. In some embodiments, n3 is 1 . In some embodiments n3 is
• n3 is 3. In some embodiments n3 is 4.
• each R 4 is independently a linear, branched, and/or cyclic Cn6 alkylenyl, alkenylenyl and/or alkynylenyl, wherein any carbon bonded to two other carbons is optionally independently replaced by N, S, or O, and carbons are optionally independently substituted.
• each n6 is independently 1 -15 or 1 -10.
• each n6 is independently 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• each R 4 is independently a C n 6 alkylenyl wherein any carbon bonded to two other carbons is optionally independently replaced by N, S, or O, and carbons are optionally independently substituted; in some such embodiments, n3 is 1. In some embodiments, n3 is 1 and R 4 is C1-C5 alkylenyl, optionally methylene. In some embodiments, each R 4 is independently -C(R aa )H-, wherein each R aa is independently the sidechain of a proteinogenic amino acid orthe sidechain of an alpha amino acid from Table 1 .
• each R 4 is independently a proteinogenic amino acid or an amino acid from Table 1 omitting the backbone amino and carboxylic acid groups of the amino acid.
• each L 4 is independently -N(R L4a )-C(O)- or - C(O)-N(R L4a )- wherein each R L4a is independently H, methyl, or ethyl.
• each R L4a is independently H or methyl.
• at least one R L4a is ethyl.
• each L 4 is independently -NHC(O)- or -C(0)-NH- In some embodiments, each L 4 is-NHC(O)-. In some embodiments, at least one L 4 is -S-. In some embodiments, at least one L 4 is -NH-C(O)-NH-. In some embodiments, at least one L 4 is -NH-C(S)-NH-. In some embodiments, at least one L 4 is In some embodiments, at least one L 4 is
• At least one L 4 is In some embodiments, at least one L 4 is
• n3 is 1
• R 4 is methylene and L 4 is -NHC(O)-.
• L 3 is -NHC(O)-.
• n2 is 1.
• each L 4 is independently -N(R L3a )-C(O)- or - C(O)-N(R L3a )- wherein each R L3a is independently H, methyl, or ethyl. In other such embodiments, each R L4a is independently H or methyl. In other such embodiments, at least one R L3a is ethyl.
• X br is a branching atom, i.e. the point where the linker diverges from a single chain to two chains to connect both R rad and R alb to the PSMA-binding moiety.
• X br is N, C or CH.
• X br is CH.
• X br is N.
• X br is separated from ring A by at least 4 ms, at least 5 atoms, at least 6 atoms, at least 7 atoms, at least 8 atoms, at least 9 atoms, or at least 10 atoms.
• the expression “X br is separated from ring A by at least [number] atoms” refers to the number of atoms that form a contiguous chain by the shortest route between X br and ring A, and excluding X br and ring A atoms from the atom count.
• the expression “by the shortest route” in this context refers to the possibility for a ring to be included in the atoms separating X br and ring A, such that there are two or more non-equivalent routes to count atoms in a contiguous chain ; in such a situation, the shortest route is counted.
• the number of atoms separating X br and ring A does not include hydrogens and does not include any non-hydrogen atoms branching off the shortest route.
• the number of atoms separating X br and ring A is 6, and excludes the two amide oxygens and excludes all hydrogens.
• X br is separated from ring A by 6 atoms. In some embodiments, X br is separated from ring A by 5 atoms. In some embodiments, X br is separated from ring A by 4 atoms.
• R 5a -X br (R 5c )-R 5b forms a C n ? alkylenyl, alkenylenyl and/or alkynylenyl, wherein n7 is 1 -20 (i.e. 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, or 20), wherein any carbon bonded to two other carbons may be independently replaced by N, S, or O heteroatoms.
• R 5a -X br (R 5c )-R 5b forms a C n ? alkylenyl.
• n7 is 1 -15 or 1 -10.
• 1 carbon is replaced by N, S, or O. In other embodiments, 2, 3, 4, or 5 carbons are each independently replaced by N, S, or O. In some embodiments, the carbons are unsubstituted. In other embodiments, 1 carbon is substituted. In other embodiments, 2 or 3 carbons are independently substituted.
• the substitutions are as defined in Formula I. In some embodiments, the substitutions are independently selected from one or more than one of hydroxyl, sulfhydryl, amine, guanidino, and/or carboxylic acid.
• R 5a is absent. In some embodiments, R 5b is absent. In some embodiments, R 5c is absent. In some embodiments, R 5a and R 5b are absent.
• R 5a and R 5c are absent. In some embodiments, R 5b and R 5c are absent. In some embodiments, all three of R 5a , R 5b , and R 5c are absent.
• R 5a is Ci-C 6 alkylenyl, optionallly — (CH 2 )I-4— . In some embodiments, R 5b is Ci-C 6 alkylenyl, optionallly -(CH 2 )I- 4 - In some embodiments, R 5c is Ci-C 6 alkylenyl, optionallly -(CH 2 )I- 4 -
• R 5a is absent or Ci-Ce alkylenyl, optionally -(CH 2 )I-4-;
• R 5b is absent or Ci-C 6 alkylenyl, optionally -(CH 2 )I_ 4 -;
• R 5C is absent or Ci-C 6 alkylenyl, optionally -(CH 2 )I_ 4 -;
• X br is N, C or CH; wherein any of R 5a , R 5b or R 5c is absent.
• R 5a -X br (R 5c )-R 5b forms: , wherein each of n12a, n12b, and n12c is independently 0-4.
• R 5a -X br (R 5c )-R 5b forms: , wherein each of n12a, n12b, and n12c is independently 0-4.
• L 5b is -N(R L5b )-C(0)- or-C(0)-N(R L5b )- wherein R L5b is H, methyl, or ethyl. In some such embodiments, R L5b is H. In other such embodiments, R L5b is methyl. In other such embodiments, R L5b is ethyl. In some embodiments, L 5b is -NHC(O)-. In some embodiments, L 5b is -C(O)-NH- In some embodiments, L 5b is -S-.
• L 5b is -N(R L3a )-C(O)- or-C(0)-N(R L3a )- wherein R L3a is H, methyl, or ethyl. In some such embodiments, R L3a is H. In other such embodiments, R L3a is methyl. In other such embodiments, R L3a is ethyl. In some embodiments, L 5b is -NHC(O)-.
• L 5b is -NH-C(O)-NH-. In some embodiments, L 5b is -NH-C(S)-NH-
• L 5b is . In some embodiments, L 5b is In some embodiments, L 5b is N-N
• L 5c is -N(R L5c )-C(0)- or-C(0)-N(R L5c )- wherein R L5c is H, methyl, or ethyl. In other such embodiments, R L5c is methyl. In other such embodiments, R L5c is ethyl. In some embodiments, L 5c is -NHC(O)-. In some embodiments, L 5c is -C(O)-NH- In some embodiments, L 5c is -S-.
• L 5c is -N(R L3a )-C(0)- or-C(0)-N(R L3a )- wherein R L3a is H, methyl, or ethyl. In other such embodiments, R L3a is methyl. In other such embodiments, R L3a is ethyl. In some embodiments, L 5c is -NHC(O)-. In some embodiments, L 5c is -C(O)-NH- In some embodiments, L 5c is -S-. In some embodiments, L 5c is -NH-C(O)-NH-
• L 5c is -NH-C(O)-NH- In some embodiments, L 5c is -NH-C(S)- NH-.
• L 5c is . In some embodiments, L 5c is some embodiments, L 5c is In some embodiments, L 5c is N-N
• n3 is 0; R 5a is methylene; X br is N or CH; and R 5b is -(CH 2 ) 4 - and R 5c is absent, or R 5c is -(CH 2 ) 4 - and R 5b is absent.
• L 5b is -NHC(O)-.
• L 5c is -NHC(O)-.
• L 5b and L 5c may each be -NHC(O)-.
• n4 is 0. In some embodiments, n4 is 1 . In some embodiments n4 is 2. In some embodiments, n4 is 3. In some embodiments n4 is 4.
• each R 6 is independently a linear, branched, and/or cyclic Cn8a alkylenyl, alkenylenyl and/or alkynylenyl, wherein each n8a is independently 1 -20, wherein any carbon bonded to two other carbons is optionally independently replaced by N, S, or O, and carbons are optionally independently substituted .
• each n8a is independently 1 -15 or 1-10.
• each n8a is independently 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• each R 6 is independently a C n sa alkylenyl wherein any carbon bonded to two other carbons is optionally independently replaced by N, S, or O, and carbons are optionally independently substituted; in some such embodiments, n4 is 1. In some embodiments, n4 is 1 and R 6 is C1-C5 alkylenyl, optionally methylene or -(CH 2 )i-4- In some embodiments, each R 6 is independently -C(R aa )H-, wherein each R aa is independently the sidechain of a proteinogenic amino acid or the sidechain of an alpha amino acid from Table 1 . In some embodiments, each R 6 is independently a proteinogenic amino acid or an amino acid from Table 1 omitting the backbone amino and carboxylic acid groups of the amino acid.
• each L 6 is independently -N(R L6a )-C(O)- or - C(O)-N(R L6a )- wherein each R L6a is independently H, methyl, or ethyl. In other such embodiments, each R L6a is independently H or methyl. In some embodiments where n4 is not zero, each L 6 is independently -N(R L3a )-C(0)- or-C(0)-N(R L3a )- wherein each R L3a is independently H, methyl, or ethyl. In other such embodiments, each R L3a is independently H or methyl.
• At least one R L6a is ethyl.
• each L 6 is independently -NHC(O)- or-C(0)-NH-
• each L 6 is-NHC(O)-.
• at least one L 6 is -S-.
• at least one L 6 is -NH-C some embodiments, L 6 is -
• At least one L 6 i is In some embodiments, at least one L 6 is In some embodiments, at least one L 6 is . In some embodiments, at least one L 6 is N-N
• n5 is 0. In some embodiments, n5 is 1 . In some embodiments n5 is 2. In some embodiments, n5 is 3. In some embodiments n5 is 4.
• each R 7 is independently a linear, branched, and/or cyclic Cn8b alkylenyl, alkenylenyl and/or alkynylenyl, wherein each n8b is independently 1 -20, wherein any carbon bonded to two other carbons is optionally independently replaced by N, S, or O, and carbons are optionally independently substituted .
• each n8b is independently 1 -15 or 1-10.
• each n8b is independently 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• each R 7 is independently a C n sb alkylenyl wherein any carbon bonded to two other carbons is optionally independently replaced by N, S, or O, and carbons are optionally independently substituted; in some such embodiments, n5 is 1. In some embodiments, n5 is 1 and R 7 is C1-C5 alkylenyl, optionally methylene or -(CH 2 )i-4- In some embodiments, each R 7 is independently -C(R aa )H-, wherein each R aa is independently the sidechain of a proteinogenic amino acid or the sidechain of an alpha amino acid from Table 1 . In some embodiments, each R 6 is independently a proteinogenic amino acid or an amino acid from Table 1 omitting the backbone amino and carboxylic acid groups of the amino acid.
• each L 7 is independently -N(R L7a )-C(O)- or - C(O)-N(R L7a )- wherein each R L7a is independently H, methyl, or ethyl. In other such embodiments, each R L7a is independently H or methyl. In other such embodiments, at least one R L7a is ethyl. In some embodiments, each L 7 is independently -NHC(O)- or -C(0)-NH- In some embodiments, each L 7 is-NHC(O)-.
• each L 7 is independently - N(R L3a )-C(0)- or-C(0)-N(R L3a )- wherein each R L3a is independently H, methyl, or ethyl. In other such embodiments, each R L3a is independently H or methyl. In other such embodiments, at least one R L3a is ethyl. In some embodiments, each L 7 is independently -NHC(O)- or -C(O)-NH- In some embodiments, each L 7 is-NHC(O)-. In some embodiments, at least one L 7 is -S-. In some embodiments, L 7 is -NH-C(O)-NH.
• L 7 is -NH-C(S)-NH-. In some embodiments, at least one L 7 is In some embodiments, at least one L 7 is In some embodiments, at least one L 7 is In some embodiments, at least one L 7 is in some embodiments, at least one L 7
• R rad is a radiometal chelator that is separated from ring A by at least 7 atoms.
• the expression “R rad is separated from ring A by at least [number] atoms” refers to the number of atoms that form a contiguous chain by the shortest route between R rad and ring A, and excluding R rad and ring A atoms from the atom count.
• the expression “by the shortest route” in this context refers to the possibility for a ring to be included in the atoms separating R rad and ring A, such that there are two or more non-equivalent routes to count atoms in a contiguous chain; in such a situation, the shortest route is counted.
• the number of atoms separating R rad and ring A does not include hydrogens and does not include any non-hydrogen atoms branching off the shortest route.
• the number of atoms separating R rad and ring A is 13 (including the linking amide attached to DOTA), and excludes the three amide oxygens, excludes the branch of the linker connecting R alb , and excludes all hydrogens.
• R rad is separated from ring A by at least 7 atoms, at least 8 atoms, at least 9 atoms, at least 10 atoms, at least 11 atoms, at least 12 atoms, at least 13 atoms, at least 14 atoms, or at least 15 atoms. In some embodiments, R rad is separated from ring A by 7- 18 atoms (e.g. 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, or 18 atoms). In some embodiments, R rad is bound by a radiometal. In some embodiments, the radiometal is not bound to R rad .
• R 3 is: . wherein R L " is
• L 2 is-N(R L2a )-C(O)-, or-C(0)-N(R L2a )-, wherein R L2a is H; and R 1 is -R 1a R 1b -, wherein R 1a is- CH 2 -, and R 1b is -CH 2 - or -CHF-.
• R 5c is absent; L 5c is -N(R L3a )-C(O)- or -C(0)-N(R L3a )-, wherein R L3a is H; R 7 is methylene; L 7 is -N(R L3a )-C(O)- or -C(0)-N(R L3a )-, wherein R L3a is H; n5 is 1 ; and R alb is: wherein n11 is 3 and R 8 is OCH 3 , or NO 2
• n2 is 0 or 1 ;
• L 3 is -N(R L3a )-C(O)-, or -C(O)-N(R L3a )-, wherein R L3a is H;
• R 4 is methylene;
• L 4 is -N(R L3a )-C(O)-, or-C(0)-N(R L3a )-, wherein R L3a is H;
• n3 is 1 ;
• R 5a is absent; and
• X br is CH.
• R 5b is a linear Ci.C 6 alkylenyl; L 5b is -N(R L3a )- C(O)- or -C(O)-N(R L3a )-, wherein R L3a is H; R 6 is methylene; L 6 is -N(R L3a )-C(O)- or -C(O)- N(R L3a )-, wherein R L3a is H; and n4 is 0-2.
• R rad is selected from Table 2, wherein R rad is optionally bound by a radiometal.
• the radiometal chelator may be any radiometal chelator suitable for binding to the radiometal and which is functionalized for attachment to an amino group. Many suitable radiometal chelators are known, e.g. as summarized in Price and Orvig, Chem. Soc. Rev., 2014, 43, 260-290, which is incorporated by reference in its entirety.
• radiometal chelators include chelators selected from the group consisting of: DOTA and derivatives; DOTAGA; NOTA; NODAGA; NODASA; CB-DO2A; 3p-C-DEPA; TCMC; DO3A; DTPA and DTPA analogues optionally selected from CHX-A”-DTPA and 1 B4M-DTPA; TETA; NOPO; Me-3,2-HOPO; CB- TE1A1 P; CB-TE2P; MM-TE2A; DM-TE2A; sarcophagine and sarcophagine derivatives optionally selected from SarAr, SarAr-NCS, diamSar, AmBaSar, and BaBaSar; TRAP; AAZTA; DATA and DATA derivatives; H2-macropa or a derivative thereof; H2dedpa, P octapa, H4py4pa, P Pypa, H 2 azapa, H 5 de
• radiometal chelators and example radioisotopes chelated by these chelators are shown in Table 2.
• the functional groups for linkage are shown in their non-linked forms; the person of skill in the art would appreciate that once linked to the compounds disclosed herein, these linking functional groups would be modified (e.g. COOH or NH 2 in the chelator would become an amide linkage when reacted with NH 2 or COOH, respectively, in the linker).
• the linkage atoms or linkageforming functional groups in Table 2 are not included in the atom count.
• R rad comprises a radiometal chelator selected from those listed above or in Table 2 linked via a linkage-forming functional group (e.g. COOH, NH 2 , SH, and the like).
• a linkage-forming functional group e.g. COOH, NH 2 , SH, and the like.
• One skilled in the art could replace any of the chelators listed herein with another chelator.
• TABLE 2 Exemplary chelators and exemplary isotopes which bind said chelators.
• the chelator is DOTA, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2.
• the chelator is CB-DO2A, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is TCMC, or a derivative thereof, linked via an amide (e.g. formed from one of the -CONH2 groups shown in Table 2).
• the chelator is 3p-C-DEPA, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is p- NH2-Bn-Oxo-D03A or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is TETA, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is CB-TE2A, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is Diamsar, or a derivative thereof, linked via an amide (e.g.
• the chelator is NOTA, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is NETA, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is HxTSE, or a derivative thereof, linked via an amide (e.g. formed from one of the amino groups shown in Table 2).
• the chelator is P2N2Ph2, or a derivative thereof, linked via an amide (e.g. formed from one of the amino groups shown in Table 2).
• the chelator is DTPA, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is CHX-AOO-DTPA, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is H2dedpa, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is H 2 azapa, or a derivative thereof, linked via an amdie (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is H 4 octapa, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is H 6 phospa, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is H 4 CHXoctapa, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is H 5 decapa, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is H 4 neunpa-p-Bn-NO2, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is SHBED, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is BPCA, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is PCTA, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is H2-MACROPA, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the chelator is Crown, or a derivative thereof, linked via an amide (e.g. formed from one of the carboxyl groups shown in Table 2).
• the radiometal chelator is a derivative of a radiometal chelator shown in Table 2.
• a derivative may include, e.g. (1) modification of a functional group of the chelator (e.g. a carboxyl group, an amino group, etc.) or (2) attachment of a new functional group (e.g. attachment of an R-group to an ethylene carbon located between two nitrogen atoms, wherein the R-group is a functional group fused to a spacer) .
• a carboxyl functional group shown in Table 2 is replaced with azidopropyl ethylacetamide (e.g. azido-mono- amide-DOTA), butynylacetamide (e.g.
• butyne-DOTA thioethylacetamide
• maleimidoethylacetamide e.g. maleimido-mono-amide-DOTA
• N-hydroxysuccinimide ester e.g. DOTA-NHS-ester
• these derivative chelators can be linked either via an amide (formed from a remaining carboxyl group) or via -C(0)-NH-(CH 2 )2-3-(triazole) or -C(O)-NH- (CH2)2-3-(thiomaleimide).
• a backbone carbon e.g.
• p-SCN-Bn-DOTA S-2-(4- isothiocyanatobenzyl)-1 ,4,7, 10-tetraazacyclododecane tetraacetic acid
• p-SCN-Bn-NOTA (2-S-(4- isothiocyanatobenzyl)-1 ,4,7-triazacyclononane-1 ,4,7-triacetic acid)
• these derivatives can form a urea linkage (formed from isocyanate) or a thiourea linkage (formed from isothiocyanate).
• the radiometal chelator is conjugated with a radioisotope (i.e. radiometal).
• the conjugated radioisotope may be, without limitation, 165 Er, 212 Bi, 166 Ho, 149 Pm, 159 Gd, 105 Rh, 109 Pd, 198 Au, 199 Au, 175 Yb, 142 Pr, 177 Lu, 111 In, 213 Bi, 212 Pb, 47 Sc, "Y, 225 Ac, 117m Sn, 153 Sm, 149 Tb, 161 Tb, 224 Ra, 227 Th, 223 Ra, 64 Cu, 67 Cu, and the like.
• the chelator is a chelator from Table 2 and the conjugated radioisotope is a radioisotope indicated in Table 2 as a binder of the particular chelator.
• the chelator is: DOTA or a derivative thereof, optionally conjugated with 177 Lu, 111 ln, 213 Bi, 212 Pb, 47 Sc, "Y, 225 Ac, 117m Sn, 153 Sm, 149 Tb, 161 Tb, 165 Er, 224 Ra, 212 Bi, 227 Th, 223 Ra, 64 Cu or 67 Cu; Crown optionally conjugated with 225 Ac, 227 Th or 177 Lu; H2- MACROPA optionally conjugated with 225 Ac; Me-3,2-HOPO optionally conjugated with 227 Th; H4py4pa optionally conjugated with 225 Ac; P pypa optionally conjugated with 177 Lu; or DTPA optionally conjugated with 111 In.
• the chelator is TETA (1 ,4,8,1 1-tetraazacyclotetradecane- 1 ,4, 8, 1 1 -tetraacetic acid), SarAr (1 -N-(4-Aminobenzyl)-3,6,10,13,16,19-hexaazabicyclo[6.6.6]- eicosane-1 ,8-diamine), NOTA (1 ,4,7-triazacyclononane-1 ,4,7-triacetic acid), TRAP (1 ,4,7- triazacyclononane-1 ,4,7-tris[methyl(2-carboxyethyl)phosphinic acid), HBED (N,N’-bis(2- hydroxybenzyl)-ethylenediamine-N,N’-diacetic acid), 2,3-HOPO (3-hydroxypyridin-2-one), PCTA (3,6,9, 15-tetraazabicyclo[9.3.1 ]-pentadeca-1 (15
• the radiometal chelator is mercaptoacetyl, hydrazinonicotinamide, dimercaptosuccinic acid, 1 ,2-ethylenediylbis-L-cysteine diethyl ester, methylenediphosphonate, hexamethylpropyleneamineoxime, or hexakis(methoxy isobutyl isonitrile).
• the chelator is bound by a radioisotope.
• the radioisotope is 186 Re or 188 Re.
• the chelator is not bound by a radioisotope.
• R alb is an albumin binder.
• R alb is -(CH 2 )n9-CH 3 wherein n9 is 8-20; in alternative embodiments, n9 is 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• R alb is -(CH 2 ) n io-C(0)OH wherein n10 is 8-20; in alternative embodiments, n10 is 8,
• R alb is separated from ring A by at least 7 atoms.
• R alb is separated from ring A by at least 4 atoms, at least 5 atoms, at least 6 atoms, at least 7 atoms, at least 8 atoms, at least 9 atoms, or at least 10 atoms.
• the expression “R alb is separated from ring A by at least [number] atoms” refers to the number of atoms that form a contiguous chain by the shortest route between R alb and ring A, and excluding R alb and ring A atoms from the atom count.
• the expression “by the shortest route” in this context refers to the possibility for a ring to be included in the atoms separating R alb and ring A, such that there are two or more non-equivalent routes to count atoms in a contiguous chain; in such a situation, the shortest route is counted.
• the number of atoms separating R alb and ring A does not include hydrogens and does not include any non-hydrogen atoms branching off the shortest route.
• the number of atoms separating R alb and ring A is 12, and excludes the four amide oxygens, excludes the branch of the linkerthat links R rad , and excludes all hydrogens.
• R alb is separated from ring A by 7-18 atoms
• R rad is separated from ring A by 7-18 atoms.
• R alb and R rad are each separated from ring A by 10-1 1 atoms.
• X br is separated from ring A by 4-6, optionally 6 atoms.
• the compound (of Formula I) has Formula II or is a salt or a solvate of Formula II: defined in Formula I, or as defined in any other embodiment(s) defined herein.
• R 1 is -CH 2 -CH 2 - or-CHF-.
• R 2 is-(CH 2 ) 4 -
• L 1 is -NH-C(O)-.
• L 2 is -NHC(O)-.
• n1 is O; ring A has 0 double bonds and is bonded at para position, optionally wherein ring some of these embodiments, n2 is 0 or 1.
• each R L3a is H.
• X br is N, C, or CH.
• R 5b is -(CH 2 ) 4 - and R 5c is absent; or R 5c is -(CH 2 ) 4 - and R 5b is absent.
• L 5b is -NH-C(O)-.
• L 5c is -NH-C(O)-.
• L 5b is-NH-C(O)-.
• n4 is 0 or 1
• R 6 — when present — is methylene.
• n5 is 0 or 1
• R 7 — when present — is methylene.
• L 6 — when present — is -NH-C(O)-.
• L 7 — when present — is -NH-C(O)-.
• R alb is wherein n1 1 is 3 and R 8 is OCH 3 or NO 2 .
• R rad is DOTA or a DOTA derivative.
• the compound is CCZ02009, CCZ02017, CCZ02008, CCZ02025, CCZ02024, CCZ02015, CCZ02019, CCZ02012, or CCZ02013, (see Examples for chemical structures) or a salt or solvate thereof, optionally conjugated with 177 Lu, 111 In, 213 Bi, 212 Pb, 47 Sc, "Y, 225 Ac, 117m Sn, 153 Sm, 161 Tb, 165 Er, 224 Ra, 212 Bi, 227 Th, 223 Ra, 64 Cu or 67 Cu.
• the compound is CCZ02009, CCZ02017, CCZ02008, CCZ02025, CCZ02024, CCZ02015, CCZ02019, CCZ02012, CCZ02005, CCZ02021 , CCZ02022, CCZ02059, CCZ02060, CCZ02034, CCZ02061 , or CCZ02013, (see Examples for chemical structures) or a salt or solvate thereof, optionally conjugated with 177 Lu, 111 In, 213 Bi, 212 Pb, 47 Sc, "Y, 225 Ac, 117m Sn, 153 Sm, 149 Tb, 161 Tb, 165 Er, 224 Ra, 212 Bi, 227 Th, 223 Ra, 64 Cu or 67 Cu.
• the compound is CCZ02005, CCZ02021 , CCZ02022, CCZ02059, CCZ02060, CCZ02034, CCZ02061 , or a salt or solvate thereof, optionally conjugated with 177 Lu, 111 ln, 213 Bi, 212 Pb, 47 Sc, "Y, 225 Ac, 117m Sn, 153 Sm, 149 Tb, 161 Tb, 165 Er, 224 Ra, 212 Bi, 227 Th, 223 Ra, 64 Cu or 67 Cu.
• radiometal chelator When the radiometal chelator is conjugated with a therapeutic radioisotope (e.g. 165 Er, 212 Bi, 166 Ho, 149 Pm, 159 Gd, 105 Rh, 109 Pd, 198 Au, 199 Au, 175 Yb, 142 Pr, 177 Lu, 111 In, 213 Bi, 212 Pb, 47 Sc, "Y, 225 Ac, 117m Sn, 153 Sm, 149 Tb, 161 Tb, 224 Ra, 227 Th, 223 Ra, 64 Cu, 67 Cu, orthe like), there is disclosed the use of certain embodiments of the compound (or a pharmaceutical composition thereof) for the treatment of PSMA-expressing conditions or diseases (e.g.
• a therapeutic radioisotope e.g. 165 Er, 212 Bi, 166 Ho, 149 Pm, 159 Gd, 105 Rh, 109 Pd, 198 Au, 199 Au, 175 Yb, 142 Pr, 177 Lu
• the compound in preparation of a medicament for treating a PSMA-expressing condition or disease in a subject.
• a method of treating PSMA-expressing disease in a subject in which the method comprises: administering to the subject a composition comprising the compound and a pharmaceutically acceptable excipient.
• the disease may be a PSMA-expressing tumor or a PSMA- expressing cancer.
• the PSMA- expressing cancer may be prostate cancer, renal cancer, breast cancer, thyroid cancer, gastric cancer, colorectal cancer, bladder cancer, pancreatic cancer, lung cancer, liver cancer, brain tumor, melanoma, neuroendocrine tumor, ovarian cancer or sarcoma.
• the cancer is prostate cancer.
• peptides which may be synthesized by any of a variety of methods established in the art. This includes but is not limited to liquid -phase as well as solid-phase peptide synthesis using methods employing 9-fluorenylmethoxycarbonyl (Fmoc) and/or t-butyloxycarbonyl (Boc) chemistries, and/or other synthetic approaches.
• Fmoc 9-fluorenylmethoxycarbonyl
• Boc t-butyloxycarbonyl
• peptides may be synthesized by sequential incorporation of the amino acid residues of interest one at a time.
• peptide synthesis is typically initiated by attaching the C-terminal amino acid of the peptide of interest to a suitable resin.
• suitable protecting groups Prior to this, reactive side chain and alpha amino groups of the amino acids are protected from reaction by suitable protecting groups, allowing only the alpha carboxyl group to react with a functional group such as an amine group, a hydroxyl group, or an alkyl halide group on the solid support.
• the protecting group on the side chain and/or the alpha amino group of the amino acid is selectively removed, allowing the coupling of the next amino acid of interest. This process is repeated until the desired peptide is fully synthesized, at which point the peptide can be cleaved from the support and purified.
• a non-limiting example of an instrument for solid-phase peptide synthesis is the Aapptec Endeavor 90 peptide synthesizer.
• Fmoc protecting groups may be removed from the amino acid on the solid support, e.g. under mild basic conditions, such as piperidine (20-50% v/v) in DMF.
• the amino acid to be added must also have been activated for coupling (e.g. at the alpha carboxylate).
• Non-limiting examples of activating reagents include without limitation 2-(1 H-benzotriazol-1-yl)-1 ,1 ,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2-(1 H-benzotriazol-1-yl)-1 ,1 ,3,3-tetramethyluronium tetrafluoroborate (TBTU), 2-(7-Aza-1 H- benzotriazole-1-yl)-1 ,1 ,3,3-tetramethyluronium hexafluorophosphate (HATU), benzotriazole-1-yl- oxy-tris(dimethylamino)phosphoniumhexafluorophosphate (BOP), benzotriazole-1-yl-oxy- tris(pyrrolidino)phosphoniumhexafluorophosphate (PyBOP).
• HBTU 2-(1 H-benzotriazol-1-yl)-1 ,1 ,3,3-
• Racemization is minimized by using triazoles, such as 1 -hydroxy-benzotriazole (HOBt) and 1 -hydroxy-7-aza-benzotriazole (HOAt). Coupling may be performed in the presence of a suitable base, such as N,N-diisopropylethylamine (DIPEA/DIEA) and the like. For long peptides or if desired, peptide synthesis and ligation may be used. [00122] Apart from forming typical peptide bonds to elongate a peptide, peptides may be elongated in a branched fashion by attaching to side chain functional groups (e.g.
• carboxylic acid groups or amino groups either: side chain to side chain; or side chain to backbone amino or carboxylate. Coupling to amino acid side chains may be performed by any known method, and may be performed on-resin or off-resin. Non-limiting examples include: forming an amide between an amino acid side chain containing a carboxyl group (e.g. Asp, D-Asp, Glu, D-Glu, and the like) and an amino acid side chain containing an amino group (e.g.
• Non-limiting examples of selectively removable protecting groups include 2- phenylisopropyl esters (O-2-PhiPr) (e.g. on Asp/Glu) as well as 4-methyltrityl (Mtt), allyloxycarbonyl (alloc), 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene))ethyl (Dde), and 1-(4,4-dimethyl-2,6- dioxocyclohex-1-ylidene)-3-methylbutyl (ivDde) (e.g. on Lys/Orn/Dab/Dap).
• 2- phenylisopropyl esters O-2-PhiPr
• Mtt 4-methyltrityl
• alloc allyloxycarbonyl
• alloc 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylidene))ethyl
• ivDde
• O-2-PhiPr and Mtt protecting groups can be selectively deprotected under mild acidic conditions, such as 2.5% trifluoroacetic acid (TFA) in DCM.
• Alloc protecting groups can be selectively deprotected using tetrakis(triphenylphosphine)palladium(0) and phenyl silane in DCM.
• Dde and ivDde protecting groups can be selectively deprotected using 2-5% of hydrazine in DMF.
• Deprotected side chains of Asp/Glu (L- or D-forms) and Lys/Orn/Dab/Dap (L- or D-forms) can then be coupled, e.g. by using the coupling reaction conditions described above.
• Peptide backbone amides may be N-methylated (i.e. alpha amino methylated) or N- alkylated. This may be achieved by directly using Fmoc-N-methylated amino acids (or Fmoc-N- alkylated amino acids) during peptide synthesis. Alternatively, N-methylation (orN-alkylation) under Mitsunobu conditions may be performed. First, a free primary amine group is protected using a solution of 4-nitrobenzenesulfonyl chloride (Ns-CI) and 2,4,6-trimethylpyridine (collidine) in NMP.
• Ns-CI 4-nitrobenzenesulfonyl chloride
• collidine 2,4,6-trimethylpyridine
• N- methylation may then be achieved in the presence of triphenylphosphine, diisopropyl azodicarboxylate (DIAD) and methanol. Subsequently, N -deprotection may be performed using mercaptoethanol and 1 ,8-diazabicyclo[5.4.0]undec-7-ene (DBU) in NMP.
• DBU 1 ,8-diazabicyclo[5.4.0]undec-7-ene
• HATU, HOAt and DIEA may be used for coupling protected amino acids to N-methylated alpha amino groups.
• the PSMA-binding moiety e.g.
• Lys-ureido-Glu, Lys-ureido-Aad, and the like may be constructed on solid phase via the formation of a ureido linkage between the amino groups of two amino acids.
• This can be done by attaching an Fmoc-protecting amino acid (for example Fmoc- Lys(ivDde)-OH) to Wang resin using standard activation/coupling strategy (for example, Fmoc- protected amino acid (4 eq.), HATU (4 eq.) and A/,/V-diisopropylethylamine (7 eq.) in N,N- dimethylformamide).
• Fmoc-protecting amino acid for example Fmoc- Lys(ivDde)-OH
• HATU HATU
• the activation and conversion of an amino group to an isocyanate group can be achieved by reacting the amino group with phosgene or triphosgene.
• the side chain functional group of the amino acid for example ivDde on Lys
• the linker, albumin-binding motif, and/or radiolabeling group e.g. radiometal chelator and the like
• thioether (-S-) linkages can be achieved either on solid phase or in solution phase.
• the formation of thioether (-S-) linkage can be achieved by coupling between a thiol-containing compound (such as the thiol group on cysteine side chain) and an alkyl halide (such as 3-(Fmoc-amino)propyl bromide and the like) in an appropriate solvent (such as N,N-dimethylformamide and the like) in the presence of base (such as N,N-diisopropylethylamine and the like).
• a thiol-containing compound such as the thiol group on cysteine side chain
• an alkyl halide such as 3-(Fmoc-amino)propyl bromide and the like
• an appropriate solvent such as N,N-dimethylformamide and the like
• base such as N,N-diisopropylethylamine and the like.
• the reactants used are preferably in equivalent molar ratio (1 to 1), and the desired products can be purified by flash column chromatography or high performance liquid chromatography (HPLC).
• HPLC high performance liquid chromatography
• the reactions are carried out on solid phase, meaning one reactant has been attached to a solid phase, then the other reactant is normally used in excess amount (> 3 equivalents of the reactant attached to the solid phase).
• the excess unreacted reactant and reagents can be removed by sequentially washing the solid phase (resin) using a combination of solvents, such as N,N-dimethylformamide, methanol and dichloromethane, for example.
• the formation of the linkage (e.g. for L 1 , L 2 , L 3 , and the like) between a thiol group and a maleimide group can be performed using the conditions described above forthe formation of the thioether (-S-) linkage simply by replacing the alkyl halide with a maleimide-containing compounds. Similarly, this reaction can be conducted in solid phase or solution phase. If the reactions are carried out in solution phase, the reactants used are preferably in equivalent molar ratio (1 to 1), and the desired products can be purified by flash column chromatography or high performance liquid chromatography (HPLC).
• HPLC high performance liquid chromatography
• the reactions are carried out on solid phase, meaning one reactant has been attached to a solid phase, then the other reactant is normally used in excess amount (> 3 equivalents of the reactant attached to the solid phase).
• the excess unreacted reactant and reagents can be removed by sequentially washing the solid phase (resin) using a combination of solvents, such as N,N-dimethylformamide, methanol and dichloromethane, for example.
• Urea or thiourea linkages can be made from reaction of an amine group with an isocyanate or an isothiocyanate, respectively, which are common functional groups on radiometal chelators.
• the isothiocyanate functional group may be added to the radiometal chelator by reacting an amino group on the chelator with thiophosgene [i.e. C(S)CI 2 ].
• the isocyanate functional group may be added to the radiometal chelator by reacting an amino group on the chelator with phosgene [i.e. C(O)CI 2 ].
• Non-peptide moieties e.g. radiometal chelator groups, albumin-binding groups and/or linkers
• a bifunctional chelator such as 1 ,4,7,10-tetraazacyclododecane-1 ,4,7,10- tetraacetic acid (DOTA) tris(tert-butyl ester) may be activated in the presence of N- hydroxysuccinimide (NHS) and N,N'-dicyclohexylcarbodiimide (DCC) for coupling to a peptide.
• N- hydroxysuccinimide NHS
• DCC N,N'-dicyclohexylcarbodiimide
• a non-peptide moiety may be incorporated into the compound via a copper-catalyzed click reaction under either liquid or solid phase conditions. Copper-catalyzed click reactions are well established in the art.
• 2-azidoacetic acid is first activated by NHS and DCC and coupled to a peptide. Then, an alkyne-containing non-peptide moiety may be clicked to the azide- containing peptide in the presence of Cu 2+ and sodium ascorbate in water and organic solvent, such as acetonitrile (ACN) and DMF and the like. Non-peptide moieties may also be added in solution phase, which is routinely performed.
• radiometal chelators are well-known and many chelators are commercially available (e.g. from Sigma-AldrichTM/Milipore SigmaTM and others). Protocols for conjugation of radiometals to the chelators are also well known (e.g. see Examples, below).
• the desired peptide may be cleaved from the solid support using suitable reagents, such as TFA, triisopropylsilane (TIS) and water.
• suitable reagents such as TFA, triisopropylsilane (TIS) and water.
• Side chain protecting groups such as Boc, pentamethyldihydrobenzofuran-5-sulfonyl (Pbf), trityl (Trt) and tert-butyl (tBu) are simultaneously removed (i.e. deprotection).
• the crude peptide may be precipitated and collected from the solution by adding cold ether followed by centrifugation.
• Purification and characterization of the peptides may be performed by standard separation techniques, such as high performance liquid chromatography (HPLC) based on the size, charge and polarity of the peptides.
• HPLC high performance liquid chromatography
• the identity of the purified peptides may be confirmed by mass spectrometry or other similar approaches.
• HPLC columns used were a semi-preparative column (Luna C18, 5 p, 250 x 10 mm) and an analytical column (Luna C18, 5 p, 250 x 4.6 mm) purchased from Phenomenex (Torrance, CA).
• the HPLC solvents were A: H2O containing 0.1 % TFA, and B: CH 3 CN containing 0.1% TFA.
• the collected HPLC eluates containing the desired peptide were lyophilized using a Labconco (Kansas City, MO) FreeZone 4.5 Plus freeze-drier. Mass analyses were performed using a Waters (Milford, Massachusetts) LC-MS with a QDa mass detector an ESI ion source.
• C18 Sep-Pak cartridges (1 cm 3 , 50 mg) were obtained from Waters (Milford, MA).
• 68 Ga was eluted from an iThemba Labs (Somerset West, South Africa) generator, and was purified using a DGA resin column from Eichrom Technologies LLC (Lisle, IL).
• Radioactivity of 68 Ga or ⁇ Lu- labeled peptides was measured using a Capintec (Ramsey, NJ) CRC®-25R/W dose calibrator, and the radioactivity of mouse tissues collected from biodistribution studies were counted using a Perkin Elmer (Waltham, MA) Wizard22480 automatic gamma counter.
• Peptidomimetic PSMA-targeting Lys-ureido-Glu moiety was synthesized by solidphase peptide chemistry. Fmoc-Lys(ivDde)-Wang resin was swelled in CH 2 CI 2 , followed by Fmoc removal by treating the resin with 20% piperidine in DMF. To generate the isocyanate of the H- Glu(OtBu)-OtBu moiety, a solution of H-Glu(OtBu)-OtBu and diisopropylethylamine in CH 2 CI 2 was cooled to -78 °C in a dry ice/acetone bath.
• T riphosgene was dissolved in CH 2 CI 2 , and the resulting solution was added dropwise to the reaction at -78 °C. The reaction was then allowed to warm to room temperature and stirred for 30 minutes. After which the isocyanate of the H-Glu(OtBu)-OtBu solution was added to the lysine-immobilized resin and reacted for 16 h. After washing the resin with DMF, the ivDde-protecting group was removed with 2% hydrazine in DMF. Fmoc-protected amino acids were then coupled to the side chain of Lys in presence of HATU and A/,/V- diisopropylethylamine.
• DOTA-tris(f-bu)ester (2-(4,7,10-tris(2-(f-butoxy)-2-oxoehtyl)- 1 ,4,7,10)-tetraazacyclododecan-1-yl)acetic acid) was coupled.
• the peptide was then deprotected and simultaneously cleaved from the resin by treating with 95/5 trifluoroacetic acid (TFA)/triisopropylsilane (TIS) for 3-4 h at room temperature. After filtration, the peptide was precipitated by the addition of the TFA solution to cold diethyl ether. The crude peptide was purified by HPLC using the preparative column. The eluates containing the desired peptide were collected, pooled, and lyophilized.
• 68 Ga was eluted from a 68 Ge-generator with 5 mL of 0.05M HCI, collected into 2.5 mL of 12M HCI and trapped onto a DGA resin. The resin was then washed with 3 mL of 5M HCI and the purified [ 68 Ga] 3+ was eluted. 177 Lu and 225 Ac were purchased from ITM (Germany). 68 Ga, 177 Lu or 225 Ac was added to 700 pL of 2M HEPES buffer containing 15-25 nmol of peptide. The reaction mixtures were then either heated at 85-90° C for 15-30 min or microwaved for 1 min.
• each solution was purified by semi-prep HPLC followed by C18 Sep-pak purification.
• LNCaP cell line was obtained from ATCC (LNCaP clone FGC, CRL-1740). It was established from a metastatic site of left supraclavicular lymph node of human prostatic adenocarcinoma. Cells were cultured in PRMI 1640 medium supplemented with 10 % FBS, penicillin (100 U/mL) and streptomycin (100 pg/mL) at 37 °C in a humidified incubator containing 5% CO 2 . Cells grown to 80-90% confluence were then washed with sterile phosphate-buffered saline (1 x PBS pH 7.4) and trypsinization.
• the collected cells number was counted with a Hausser Scientific (Horsham, PA) Hemacytometer. Approximately 10 million LNCaP cells were inoculated into the left dorsal flank of immunocomprised NRG mice. The tumours were allowed to grow for 4-6 weeks and used for imaging and biodistribution studies when a volume of 200-600 mm 3 was reached.
• Inhibition constants (Ki) to PSMA were measured by in vitro competition binding assays using [ 18 F]DCFPyl_ as the radioligand.
• LNCaP cells which were plated onto a 24-well poly- D-lysine coated plate for 48 h (400,000/well). Growth medium was removed and replaced with HEPES buffered saline (50 mM HEPES, pH 7.5, 0.9% sodium chloride).
• [ 18 F]DCFPyL (0.1 nM) was added to each well (in triplicate) containing varied concentrations (0.5 mM - 0.05 nM) of tested compounds
• the assay mixtures were incubated for 1 h at 37 °C with gentle agitation followed by two washes with cold HEPES buffered saline.
• a trypsin solution (0.25 %, 400pL) was then added to each well to harvest the cells. Radioactivity was measured by gamma counting and Ki values calculated using the ‘one site - fit Ki’ built-in model in Prism 8 (GraphPad).
• PET imaging experiments were conducted using Siemens Inveon micro PET/CT scanner.
• SPECT imaging experiments were conducted using an MILabs micro SPECT/CT scanner.
• Each tumor bearing mouse was injected 4-6 MBq of 68 Ga or 18.5 MBq of 177 Lu labeled tracer through the tail vein under anesthesia (2% isoflurane in oxygen).
• the mice were allowed to recover and roam freely in their cage.
• the mice were sedated again with 2% isoflurane in oxygen inhalation and positioned in the scanner.
• a 10-min CT scan was conducted first for localization and attenuation correction after segmentation for reconstructing the PET or SPECT images.
• a 10-min static PET imaging or a one hour (30 min X 2 frames) of static SPECT scan was performed to determined uptake in tumor and other organs.
• the mice were kept warm by a heating pad during acquisition.
• mice were injected with the radiotracer as described above.
• the mice were anesthetized with 2% isoflurane inhalation, and euthanized by CO 2 inhalation.
• Blood was withdrawn immediately from the heart, and the organs/tissues of interest were collected.
• the collected organs/tissues were weighed and counted using a Perkin Elmer (Waltham, MA) Wizard22480 gamma counter.
• the uptake in each organ/tissue was normalized to the injected dose (radioactivity) using a standard curve, and expressed as the percentage of the injected dose per gram of tissue (%ID/g).
• EXAMPLE 1 SYNTHESIS AND EVALUATION OF CCZ02009 IN COMPARISON TO HTK03170 WITHOUT A GLY SPACER
• CCZ02009 The synthesis of CCZ02009 follows the general synthesis procedures as described above except that CCZ02009 is based on the Lys-ureido-Aad moiety.
• a solution of L-2-aminoadipic acid (Aad) di-tertbutyl ester hydrochloride and diisopropylethylamine in CH 2 CI 2 was cooled to -78 °C in a dry ice/acetone bath.
• Triphosgene was dissolved in CH 2 CI 2 , and the resulting solution was added dropwise to the reaction at -78 °C.
• HTK03170 The chemical structure of HTK03170 is shown below:
• Table 3 shows that with a Gly spacer between the PSMA and albumin binding moieties, PSMA binding affinity increased from 1 .53 ⁇ 0.33 to 0.12 ⁇ 0.02 nM, and albumin binding affinity increased from 70.1 ⁇ 3.2 to 64.9 ⁇ 2.2 pM.
• Figure 2A shows the representative binding affinity curves for CCZ02009 (left panel) and HTK03170 (right panel).
• Figure 2B and Table 4 show the PET imaging and biodistribution results using 68 Ga-CCZ02009 in NRG-mice bearing LNCaP tumors 1 h ( Figure 2B left panel) and 3 h ( Figure 2B right panel) post-injection (p.i.). Overall high and sustained blood radioactivity was observed at both timepoints, indicating the strong albumin binding of CCZ02009.
• EXAMPLE 2 SYNTHESIS AND EVALUATION OF CCZ02060and CCZ02059with additional Gly spacers
• CCZ02060 and CCZ02059 follows the synthesis procedures of CCZ02009 as described above, except that CCZ02060 contains two Gly spacers and CCZ02059 contains four Gly spacers.
• mass calculated [M+2H] 2+ 791 .4, found 791 .5.
• mass calculated [M+2H] 2+ 848.4, found 848.5.
• CCZ02060 and CCZ02059 binds to PSMA with affinities (Ki) of 1 .09 ⁇ 0.19 nM and
• Figure 3 shows the representative binding affinity curves for CCZ02060 (left panel) and CCZ02059 (right panel).
• the number of Gly spacers has an impact on the PSMA binding affinity, i.e. the most favorable binding affinity was observed with a single Gly spacer (CCZ02009), followed by two Gly spacers (CCZ02060) and four Gly spacers (CCZ02059).
• EXAMPLE 3 SYNTHESIS AND EVALUATION OF CCZ02017 IN COMPARISON TO HTK03170 WITHOUT A GLY SPACER
• CCZ02017 follows the synthesis procedures of CCZ02009 as described above, except for the albumin binder, which is 4-(p-nitrophenyl)butyric acid in CCZ02017.
• the 4-(p-nitrophenyl)butyric acid is substantially weaker than 4-(p-methoxyphenyl)butyric acid (Ref. Kuo et al. J Nucl Med. 2021 Apr;62(4):521 -527).
• Figure 4 shows SPECT/CT images of 177 Lu- CCZ02017 in NRG-mice bearing LNCaP tumors at 3, 24, 72, 144 and 240 h p.i.
• Table 5 shows the biodistribution of 177 Lu-CCZ02017 in NRG-mice bearing LNCaP tumors at 3, 24, 72, 144 and 240 h p.i.
• 177 Lu-HTK03170 shows similar blood % I D/g values (Table 4) to 177 Lu-CCZ02017 at 24 and 72 h p.i. (see Table 6). Since CCZ02017 has a weaker albumin binder, the data indicates that the introduction of a Gly spacer improves albumin binding.
• the LNCaP tumor uptake in 177 Lu-CCZ02017 is substantially improved compared to 177 Lu-HTK03170.
• EXAMPLE 4 SYNTHESIS AND EVALUATION OF CCZ02008
• EXAMPLE 6 SYNTHESIS AND EVALUATION OF CCZ02024
• EXAMPLE 7 SYNTHESIS AND EVALUATION OF CCZ02015
• EXAMPLE 8 SYNTHESIS AND EVALUATION OF CCZ02019
• EXAMPLE 9 SYNTHESIS AND EVALUATION OF CCZ02012 and CCZ02013
• CCZ02013 [00185] The synthesis of CCZ02012 and CCZ02013 follows the general synthesis procedures as described above on the Lys-ureido-Aad backbone, and instead of Fmoc-T ranexamic acid, Fmoc-trans-4-aminocyclohexane carboxylic acid (ACHC) and Fmoc-cis-4-ACHC were incorporated, respectively.
• ACHC Fmoc-trans-4-aminocyclohexane carboxylic acid
• ACHC eithertrans or cis, did not affect the binding to PSMA substantially.
• Figure 9 shows the binding assay curves for CCZ02012 (9A) and CCZ02013 (9B).
• EXAMPLE 10 SYNTHESIS AND EVALUATION OF CCZ02021 and CCZ02022
• CCZ02022 [00188] The synthesis of CCZ02021 and CCZ02022 follows the synthesis procedures of CCZ02012 as described with CCZ02021 incorporating a Gly spacer, and CCZ02022 incorporating a Gly spacer and beta-homoLys instead of Lys.
• mass calculated [M+2H] 2+ 763.4, found 763.5.
• mass calculated [M+2H] 2+ 770.4, found 770.7.
• Figure 9 shows the binding assay curves for CCZ02021 (10A) and CCZ02022 (10B).
• EXAMPLE 11 SYNTHESIS AND EVALUATION OF CCZ02034
• EXAMPLE 12 SYNTHESIS AND EVALUATION OF CCZ02005
• EXAMPLE 13 SYNTHESIS AND EVALUATION OF CCZ02061
• the present invention has been described with regard to one or more embodiments. However, it will be apparent to persons skilled in the art that a number of variations and modifications can be made without departing from the scope of the invention as defined in the following claims. The scope of the invention should therefore not be limited by the preferred embodiments set forth in the above Examples, but should be given the broadest interpretation consistent with the description as a whole.

Landscapes

• Chemical & Material Sciences (AREA)
• Health & Medical Sciences (AREA)
• Organic Chemistry (AREA)
• Life Sciences & Earth Sciences (AREA)
• Medicinal Chemistry (AREA)
• General Health & Medical Sciences (AREA)
• Public Health (AREA)
• Pharmacology & Pharmacy (AREA)
• Animal Behavior & Ethology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Veterinary Medicine (AREA)
• Epidemiology (AREA)
• Physics & Mathematics (AREA)
• Optics & Photonics (AREA)
• Biophysics (AREA)
• Molecular Biology (AREA)
• General Chemical & Material Sciences (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Biochemistry (AREA)
• Genetics & Genomics (AREA)
• Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
• Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
• Peptides Or Proteins (AREA)

Priority Applications (5)

Applications Claiming Priority (2)

Publications (1)

Family

ID=87882764

Family Applications (1)

Country Status (6)

Cited By (4)

Citations (4)

• 2023
  • 2023-03-03 CA CA3244865A patent/CA3244865A1/en active Pending
  • 2023-03-03 EP EP23762646.0A patent/EP4486756A1/en active Pending
  • 2023-03-03 WO PCT/CA2023/050280 patent/WO2023164775A1/en not_active Ceased
  • 2023-03-03 AU AU2023226956A patent/AU2023226956A1/en active Pending
  • 2023-03-03 US US18/842,681 patent/US20250186627A1/en active Pending
  • 2023-03-03 JP JP2024552478A patent/JP2025507971A/ja active Pending

Patent Citations (4)

Non-Patent Citations (2)

Also Published As

Similar Documents

Legal Events