Classifications

• • 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
  • 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
  • A61K51/041—Heterocyclic compounds
  • A61K51/044—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins
  • A61K51/0459—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine, rifamycins having six-membered rings with two nitrogen atoms as the only ring hetero atoms, e.g. piperazine
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2121/00—Preparations for use in therapy
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K2123/00—Preparations for testing in vivo

Definitions

• the present disclosure relates to new folate-conjugates comprising a 5-methyltetrahydro- folate, a radiometal chelator optionally coordinating a radiometal M, and the albumin binder 5-(p-iodophenyl)pentanoate, and further provides uses of such conjugates and/or pharmaceutical compositions thereof in diagnostic imaging, radionuclide therapy or theragnostic applications.
• FR folate receptor-alpha
• FR is a membrane-associated glycoprotein which is overexpressed on a variety of tumor types, among those ovarian, lung, breast, renal and colorectal cancer (Parker, N.
• Folic acid has, therefore, been used as a targeting agent to deliver attached diagnostic and therapeutic payloads for imaging and therapy of FR-expressing cancer (Low, P. S. et al., Acc Chem Res 2008, 41, (1)).
• Only a few of the developed folate radioconjugates were used in clinics, among those [ 111 In]In-DTPA- folate and [ 99m Tc]Tc-EC20 (Etarfolatide TM , Endocyte Inc.) for single photon emission computed tomography (SPECT) (Siegel, B. A. et al., J Nucl Med 2003, 44, (5), 700-7; Fisher, R. E.
• albumin-binding radioconjugates comprising 5-methyltetrahydrofolate (5-MTHF) as a targeting agent showed high tumor-to- kidney ratios and, as a consequence, a superior therapeutic effect as compared to the respective folic acid based compound [ 177 Lu]Lu-OxFol-1 (Guzik, P. et al., Eur J Nucl Med Mol Imaging 2021, 48, 972–983).
• the present disclosure is in a first aspect directed to new folate-conjugates comprising a 5-methyltetrahydrofolate, a radiometal chelator optionally coordinating a radiometal M, and 5-(p-iodophenyl)pentanoate as an albumin binder.
• the new folate conjugates are compounds of formula I, or a stereoisomer (or a combination of stereoisomers) thereof, or a pharmaceutically acceptable salt thereof
• the radiometal chelator is selected from linear or macrocyclic polyaminocarboxylates, such as DTPA, DOTA (and derivatives thereof, such as p-SCN- DOTA, maleimido-DOTA, DOTA-NHS-ester), DFO, DFO*, DO3A, AAZTA, HP-DO3A, EDTA, TETA, EHPG, HBED, NOTA (and derivatives such as p-SCN-NOTA), DOTAGA, DOTMA, TETMA, PDTA, TTHA, LICAM, MECAM, AAZTA, preferably macrocyclic polyaminocarboxylates, such as NOTA, DOTA, DTPA, DO3A, HP-DO3A, EDTA, TETA, DOTMA, AAZTA.
• linear or macrocyclic polyaminocarboxylates such as DTPA, DOTA (and derivatives thereof, such as p-SCN- DOTA, maleimido-DOTA, DOTA-
• the radiometal chelator e.g. the macrocyclic polyaminocarboxylate is covalently bound to a compound of the disclosure through amide coupling of one of its carboxylate groups.
• the radiometal chelator may or may not be coordinating a radiometal M.
• the optionally coordinated radiometal M is selected from 51 Cr, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 47 Sc, 167 Tm, 141 Ce, 111 In, 168 Yb, 175 Yb, 140 La, 89 Zr, 90 Y, 88 Y, 153 Sm, 166 Ho, 52 Mn, 165 Dy, 166 Dy, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 97 Ru, 103 Ru, 186 Re, 188 Re, 203 Pb, 211 Bi, 212 Bi, 213 Bi, 214 Bi, 105 Rh, 109 Pd, 212 Pb, 117m Sn, 149 Pm, 161 Tb, 149 Tb, 152 Tb, 155 Tb, 99m Tc, 165 Er, 169 Er, 172 Yb, 165 Tm, 177 Lu, 225 Ac, 198 Au, 199 Au, and 227 Th.
• the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 67 Cu, 43 Sc, 44 Sc, 99m Tc, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 90 Y, 177 Lu, and 225 Ac.
• the optionally coordinated radiometal M for use in diagnostic imaging is selected from 61 Cu, 62 Cu, 64 Cu, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 52 Mn, 89 Zr, 99m Tc, 111 In, 152 Tb, and 155 Tb.
• the optionally coordinated radiometal M for use in diagnostic imaging is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, and 155 Tb.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 105 Rh, 117m Sn, 149 Pm, 153 Sm, 161 Tb, 149 Tb, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 225 Ac, and 213 Bi.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• n is 2, 3, 4, 5 or 6, preferably 4.
• m is 2, 3, 4, 5 or 6, preferably 4.
• n is 4 and m is 4.
• the present disclosure provides uses of a compound and/or pharmaceutical composition of the present disclosure in diagnostic imaging, radionuclide therapy or theragnostic applications.
• the subject of the methods of the present disclosure is a mammal, such as an animal or a human. In some embodiments, the subject of the methods of the present disclosure is a human.
• Tumor-to-kidney ratios determined based on biodistribution data obtained at 1 h, 4 h and 24 h after injection of respective radioconjugates.
• Figure 6. Tumor-to-liver ratios determined based on biodistribution data obtained at 1 h, 4 h and 24 h after injection of respective radioconjugates.
• Figure 7. SPECT/CT images shown as maximum intensity projections (MIPs) of KB tumor- bearing mice 1 h, 4 h and 24 h after injection of the 177 Lu-folate radioconjugates (25 MBq; 0.5 nmol per mouse).
• A SPECT/CT scans of [ 177 Lu]Lu-OxFol radioconjugates ;
• B SPECT/CT scans of 6R-5-MTHF-based radioconjugates ;
• C SPECT/CT scans of 6S- 5-MTHF-based radioconjugates ;
• the present disclosure is in a first aspect directed to new folate conjugates (hereinafter also called compounds or conjugates of the disclosure) comprising a 5-methyltetrahydrofolate (5-MTHF), a radiometal chelator optionally coordinating a radiometal M, and the albumin binder 5-(p-iodophenyl)pentanoate.
• a radiometal chelator optionally coordinating a radiometal M
• albumin binder 5-(p-iodophenyl)pentanoate The term “radiometal chelator” (or (metal) chelator) may be any of the metal chelators known in the art for complexing a radiometal or radionuclide (and useful for the intended applications).
• the binding of a chelator to a radiometal may be determined by measuring the dissociation constant between chelator and radiometal.
• the dissociation constant KD between chelator and radiometal is from about 10 -3 to about 10 -15 M -1 .
• the dissociation constant KD between chelator and radiometal is from about 10 -6 to about 10 -15 M -1 .
• a radiometal for use in the present disclosure is one, which can be detected externally in a non-invasive manner following administration in vivo.
• the radiometal is particularly one which is suitable for imaging using SPECT or PET.
• chelators are well known in the art, and include bidentate, tridentate, and tetradentate ligands in linear, tripodal and macrocyclic form.
• Typical examples include bipyridyl (bipy); terpyridyl (terpy); crown ethers; aza-crown ethers; succinic acid; citric acid; salicylic acids; histidines; imidazoles; ethyleneglycol-bis-(beta-aminoethyl ether) N,N'- tetraacetic acid (EGTA); nitroloacetic acid; acetylacetonate (acac); sulfate; dithiocarbamates; carboxylates; alkyldiamines; ethylenediamine (en); diethylenetriamine (dien); nitrate; nitro; nitroso; (C6H5)2PCH2CH2P(C6H5)2 (diphos); glyme; diglyme; bis(acetylacetonate) ethylenediamine (acacen); ethylenediaminotetraacetic acid (EDTA), diethylenetriaminopen
• Suitable metal chelators for use in the compounds of the present disclosure include bidentate, tridentate, and tetradentate, ligands in linear, tripodal and macrocyclic form, as identified hereinabove.
• the metal chelators used for the present disclosure include linear or macrocyclic polyaminocarboxylates, such as DTPA, DOTA (and derivatives thereof, such as p-SCN-DOTA, maleimido-DOTA, DOTA-NHS-ester), DFO, DFO*, DO3A, HP-DO3A, AAZTA, EDTA, TETA, EHPG, HBED, NOTA (and derivatives such as p-SCN-NOTA), DOTAGA, DOTMA, TETMA, PDTA, TTHA, LICAM, MECAM.
• the metal chelators used for the present disclosure include macrocyclic polyaminocarboxylates, such as NOTA, DOTA, DTPA, DO3A, HP-DO3A, EDTA, TETA, DOTMA, AAZTA.
• the radiometal chelator as defined herein is linked to a compound of the disclosure through one of its carboxylate groups, for example through coupling with an amino group to give an amide linkage.
• the radiometal chelator may or may not be coordinating a radiometal M.
• the term “radiometal” also referred to as radionuclide refers to an atom capable of undergoing radioactive decay and may be used as diagnostic imaging agents or therapeutic agents as described below.
• radiometal for nuclear imaging or radionuclide therapy include 51 Cr, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 47 Sc, 167 Tm, 141 Ce, 111 In, 168 Yb, 175 Yb, 140 La, 89 Zr, 90 Y, 88 Y, 153 Sm, 166 Ho, 52 Mn, 165 Dy, 166 Dy, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 97 Ru, 103 Ru, 186 Re, 188 Re, 203 Pb, 211 Bi, 212 Bi, 213 Bi, 214 Bi, 105 Rh, 109 Pd, 212 Pb, 117m Sn, 149 Pm, 161 Tb, 149 Tb, 152 Tb, 155 Tb, 99m Tc, 165 Er, 169 Er, 172 Yb, 165 Tm, 177 Lu, 225 Ac, 198 Au, 199 Au, and 227 Th.
• the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 67 Cu, 43 Sc, 44 Sc, 99m Tc, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 90 Y, 177 Lu, and 225 Ac. In some embodiments, the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, 155 Tb, 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac. The choice of metal will be determined based on the intended therapeutic or diagnostic use. A skilled person will know which radiometal to choose for the intended application.
• the optionally coordinated radiometal M for use in diagnostic imaging is selected from 61 Cu, 62 Cu, 64 Cu, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 52 Mn, 89 Zr, 99m Tc, 111 In, 152 Tb, and 155 Tb. In some embodiments, the optionally coordinated radiometal M for use in diagnostic imaging is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, and 155 Tb.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 105 Rh, 117m Sn, 149 Pm, 153 Sm, 161 Tb, 149 Tb, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 225 Ac, and 213 Bi.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• compounds of the disclosure encompasses compounds of formula (I) to (III) and any stereoisomers thereof (such as explicitly the (6R)- or (6S)-isomers) and any pharmaceutically acceptable salt thereof.
• the disclosure also encompasses compounds of the disclosure, in which one or more atoms are replaced by a specific isotope of the corresponding atom, e.g. in which one or more or all hydrogen atom are replaced by deuterium atoms D to form compounds of the disclosure that are enriched in deuterium.
• a compound of the disclosure is a compound of formula I or pharmaceutically acceptable salt or a stereoisomer thereof I wherein Y is a radiometal chelator optionally coordinating a radiometal M, n is 1 to 8, and m is 1 to 8.
• the radiometal chelator is as defined hereinabove.
• the radiometal chelator is selected from linear or macrocyclic polyaminocarboxylates, such as DTPA, DOTA (and derivatives thereof, such as p-SCN- DOTA, maleimido-DOTA, DOTA-NHS-ester), DFO, DFO*, DO3A, HP-DO3A, AAZTA, EDTA, TETA, EHPG, HBED, NOTA (and derivatives such as p-SCN-NOTA), DOTAGA, DOTMA, TETMA, PDTA, TTHA, LICAM, MECAM.
• linear or macrocyclic polyaminocarboxylates such as DTPA, DOTA (and derivatives thereof, such as p-SCN- DOTA, maleimido-DOTA, DOTA-NHS-ester), DFO, DFO*, DO3A, HP-DO3A, AAZTA, EDTA, TETA, EHPG, HBED, NOTA (and derivatives such as p-SCN-
• the metal chelators used for the present disclosure include macrocyclic polyaminocarboxylates, such as NOTA, DOTA, DTPA, DO3A, HP-DO3A, AAZTA, EDTA, TETA, DOTMA.
• macrocyclic polyaminocarboxylates such as NOTA, DOTA, DTPA, DO3A, HP-DO3A, AAZTA, EDTA, TETA, DOTMA.
• the optionally coordinated radiometal M is selected from 51 Cr, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 47 Sc, 167 Tm, 141 Ce, 111 In, 168 Yb, 175 Yb, 140 La, 89 Zr, 90 Y, 88 Y, 153 Sm, 166 Ho, 52 Mn, 165 Dy, 166 Dy, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 97 Ru, 103 Ru, 186 Re, 188 Re, 203 Pb, 211 Bi, 212 Bi, 213 Bi, 214 Bi, 105 Rh, 109 Pd, 212 Pb, 117m Sn, 149 Pm, 161 Tb, 149 Tb, 152 Tb, 155 Tb, 99m Tc, 165 Er, 169 Er, 172 Yb, 165 Tm, 177 Lu, 225 Ac, 198 Au, 199 Au, and 227 Th.
• the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 67 Cu, 43 Sc, 44 Sc, 99m Tc, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 90 Y, 177 Lu, and 225 Ac. In some embodiments, the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, 155 Tb, 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• the optionally coordinated radiometal M for use in diagnostic imaging is selected from 61 Cu, 62 Cu, 64 Cu, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 52 Mn, 89 Zr, 99m Tc, 111 In, 152 Tb, and 155 Tb. In some embodiments, the optionally coordinated radiometal M for use in diagnostic imaging is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, and 155 Tb.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 105 Rh, 117m Sn, 149 Pm, 153 Sm, 161 Tb, 149 Tb, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 225 Ac, and 213 Bi.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• n is 2, 3, 4, 5 or 6, preferably 4.
• m is 2, 3, 4, 5 or 6, preferably 4.
• a compound of formula I has the formula Ia or Ib Ia Ib wherein Y is a radiometal chelator optionally coordinating a radiometal M, n is 1 to 8, and m is 1 to 8.
• the radiometal chelator is as defined hereinabove.
• the radiometal chelator is selected from linear or macrocyclic polyaminocarboxylates, such as DTPA, DOTA (and derivatives thereof, such as p-SCN- DOTA, maleimido-DOTA, DOTA-NHS-ester), DFO, DFO*, DO3A, HP-DO3A, AAZTA, EDTA, TETA, EHPG, HBED, NOTA (and derivatives such as p-SCN-NOTA), DOTAGA, DOTMA, TETMA, PDTA, TTHA, LICAM, MECAM.
• linear or macrocyclic polyaminocarboxylates such as DTPA, DOTA (and derivatives thereof, such as p-SCN- DOTA, maleimido-DOTA, DOTA-NHS-ester), DFO, DFO*, DO3A, HP-DO3A, AAZTA, EDTA, TETA, EHPG, HBED, NOTA (and derivatives such as p-SCN-
• the metal chelators used for the present disclosure include macrocyclic polyaminocarboxylates, such as NOTA, DOTA, DTPA, DO3A, HP-DO3A, AAZTA, EDTA, TETA, DOTMA.
• macrocyclic polyaminocarboxylates such as NOTA, DOTA, DTPA, DO3A, HP-DO3A, AAZTA, EDTA, TETA, DOTMA.
• the optionally coordinated radiometal M is selected from 51 Cr, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 47 Sc, 167 Tm, 141 Ce, 111 In, 168 Yb, 175 Yb, 140 La, 89 Zr, 90 Y, 88 Y, 153 Sm, 166 Ho, 52 Mn, 165 Dy, 166 Dy, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 97 Ru, 103 Ru, 186 Re, 188 Re, 203 Pb, 211 Bi, 212 Bi, 213 Bi, 214 Bi, 105 Rh, 109 Pd, 212 Pb, 117m Sn, 149 Pm, 161 Tb, 149 Tb, 152 Tb, 155 Tb, 99m Tc, 165 Er, 169 Er, 172 Yb, 165 Tm, 177 Lu, 225 Ac, 198 Au, 199 Au, and 227 Th.
• the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 67 Cu, 43 Sc, 44 Sc, 99m Tc, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 90 Y, 177 Lu, and 225 Ac. In some embodiments, the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, 155 Tb, 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• the optionally coordinated radiometal M for use in diagnostic imaging is selected from 61 Cu, 62 Cu, 64 Cu, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 52 Mn, 89 Zr, 99m Tc, 111 In, 152 Tb, and 155 Tb. In some embodiments, the optionally coordinated radiometal M for use in diagnostic imaging is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, and 155 Tb.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 105 Rh, 117m Sn, 149 Pm, 153 Sm, 161 Tb, 149 Tb, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 225 Ac, and 213 Bi.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• n is 2, 3, 4, 5 or 6, preferably 4.
• m is 2, 3, 4, 5 or 6, preferably 4.
• a compound of formula I has the formula II II wherein Y is a radiometal chelator optionally coordinating a radiometal M.
• the radiometal chelator is as defined hereinabove.
• the radiometal chelator is selected from linear or macrocyclic polyaminocarboxylates, such as DTPA, DOTA (and derivatives thereof, such as p-SCN- DOTA, maleimido-DOTA, DOTA-NHS-ester), DFO, DFO*, DO3A, HP-DO3A, AAZTA, EDTA, TETA, EHPG, HBED, NOTA (and derivatives such as p-SCN-NOTA), DOTAGA, DOTMA, TETMA, PDTA, TTHA, LICAM, MECAM.
• linear or macrocyclic polyaminocarboxylates such as DTPA, DOTA (and derivatives thereof, such as p-SCN- DOTA, maleimido-DOTA, DOTA-NHS-ester), DFO, DFO*, DO3A, HP-DO3A, AAZTA, EDTA, TETA, EHPG, HBED, NOTA (and derivatives such as p-SCN-
• the metal chelators used for the present disclosure include macrocyclic polyaminocarboxylates, such as NOTA, DOTA, DTPA, DO3A, HP-DO3A, AAZTA, EDTA, TETA, DOTMA.
• macrocyclic polyaminocarboxylates such as NOTA, DOTA, DTPA, DO3A, HP-DO3A, AAZTA, EDTA, TETA, DOTMA.
• the optionally coordinated radiometal M is selected from 51 Cr, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 47 Sc, 167 Tm, 141 Ce, 111 In, 168 Yb, 175 Yb, 140 La, 89 Zr, 90 Y, 88 Y, 153 Sm, 166 Ho, 52 Mn, 165 Dy, 166 Dy, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 97 Ru, 103 Ru, 186 Re, 188 Re, 203 Pb, 211 Bi, 212 Bi, 213 Bi, 214 Bi, 105 Rh, 109 Pd, 212 Pb, 117m Sn, 149 Pm, 161 Tb, 149 Tb, 152 Tb, 155 Tb, 99m Tc, 165 Er, 169 Er, 172 Yb, 165 Tm, 177 Lu, 225 Ac, 198 Au, 199 Au, and 227 Th.
• the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 67 Cu, 43 Sc, 44 Sc, 99m Tc, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 90 Y, 177 Lu, and 225 Ac. In some embodiments, the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, 155 Tb, 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• the optionally coordinated radiometal M for use in diagnostic imaging is selected from 61 Cu, 62 Cu, 64 Cu, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 52 Mn, 89 Zr, 99m Tc, 111 In, 152 Tb, and 155 Tb. In some embodiments, the optionally coordinated radiometal M for use in diagnostic imaging is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, and 155 Tb.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 105 Rh, 117m Sn, 149 Pm, 153 Sm, 161 Tb, 149 Tb, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 225 Ac, and 213 Bi.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• a compound of formula I has the formula IIa or IIb IIa
• Y is a radiometal chelator optionally coordinating a radiometal M.
• the radiometal chelator is as defined hereinabove.
• the radiometal chelator is selected from linear or macrocyclic polyaminocarboxylates, such as DTPA, DOTA (and derivatives thereof, such as p-SCN- DOTA, maleimido-DOTA, DOTA-NHS-ester), DFO, DFO*, DO3A, HP-DO3A, AAZTA, EDTA, TETA, EHPG, HBED, NOTA (and derivatives such as p-SCN-NOTA), DOTAGA, DOTMA, TETMA, PDTA, TTHA, LICAM, MECAM.
• the metal chelators used for the present disclosure include macrocyclic polyaminocarboxylates, such as NOTA, DOTA, DTPA, DO3A, HP-DO3A, AAZTA, EDTA, TETA, DOTMA.
• macrocyclic polyaminocarboxylates such as NOTA, DOTA, DTPA, DO3A, HP-DO3A, AAZTA, EDTA, TETA, DOTMA.
• the optionally coordinated radiometal M is selected from 51 Cr, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 47 Sc, 167 Tm, 141 Ce, 111 In, 168 Yb, 175 Yb, 140 La, 89 Zr, 90 Y, 88 Y, 153 Sm, 166 Ho, 52 Mn, 165 Dy, 166 Dy, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 97 Ru, 103 Ru, 186 Re, 188 Re, 203 Pb, 211 Bi, 212 Bi, 213 Bi, 214 Bi, 105 Rh, 109 Pd, 212 Pb, 117m Sn, 149 Pm, 161 Tb, 149 Tb, 152 Tb, 155 Tb, 99m Tc, 165 Er, 169 Er, 172 Yb, 165 Tm, 177 Lu, 225 Ac, 198 Au, 199 Au, and 227 Th.
• the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 67 Cu, 43 Sc, 44 Sc, 99m Tc, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 90 Y, 177 Lu, and 225 Ac. In some embodiments, the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, 155 Tb, 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• the optionally coordinated radiometal M for use in diagnostic imaging is selected from 61 Cu, 62 Cu, 64 Cu, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 52 Mn, 89 Zr, 99m Tc, 111 In, 152 Tb, and 155 Tb. In some embodiments, the optionally coordinated radiometal M for use in diagnostic imaging is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, and 155 Tb.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 105 Rh, 117m Sn, 149 Pm, 153 Sm, 161 Tb, 149 Tb, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 225 Ac, and 213 Bi.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• a compound of formula I has the formula III and is optionally coordinating a radiometal M III
• the optionally coordinated radiometal M is selected from 51 Cr, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 47 Sc, 167 Tm, 141 Ce, 111 In, 168 Yb, 175 Yb, 140 La, 89 Zr, 90 Y, 88 Y, 153 Sm, 166 Ho, 52 Mn, 165 Dy, 166 Dy, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 97 Ru, 103 Ru, 186 Re, 188 Re, 203 Pb, 211 Bi, 212 Bi, 213 Bi, 214 Bi, 105 Rh, 109 Pd, 212 Pb, 117m Sn, 149 Pm, 161 Tb, 149 Tb, 152 Tb, 155 Tb, 99m Tc, 165 Er, 169 Er, 172 Yb, 165 Tm, 177 Lu, 225 Ac
• the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 67 Cu, 43 Sc, 44 Sc, 99m Tc, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 90 Y, 177 Lu, and 225 Ac. In some embodiments, the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, 155 Tb, 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• the optionally coordinated radiometal M is for use in diagnostic imaging and is selected from 61 Cu, 62 Cu, 64 Cu, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 52 Mn, 89 Zr, 99m Tc, 111 In, 152 Tb, and 155 Tb. In some embodiments, the optionally coordinated radiometal M is for use in diagnostic imaging and is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, and 155 Tb.
• the optionally coordinated radiometal M is for use in radionuclide therapy and is selected from 67 Cu, 90 Y, 105 Rh, 117m Sn, 149 Pm, 153 Sm, 161 Tb, 149 Tb, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 225 Ac, and 213 Bi.
• the optionally coordinated radiometal M is for use in radionuclide therapy and is selected from 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• a compound of formula I has the formula IIIa or IIIb and is optionally coordinating a radiometal M IIIa
• the optionally coordinated radiometal M is selected from 51 Cr, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 47 Sc, 167 Tm, 141 Ce, 111 In, 168 Yb, 175 Yb, 140 La, 89 Zr, 90 Y, 88 Y, 153 Sm, 166 Ho, 52 Mn, 165 Dy, 166 Dy, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 97 Ru, 103 Ru, 186 Re, 188 Re, 203 Pb, 211 Bi, 212 Bi, 213 Bi, 214 Bi, 105 Rh, 109 Pd, 212 Pb, 117m Sn, 149 Pm, 161 Tb, 149 Tb, 152 Tb, 155 Tb, 99m Tc, 165 Er, 169 Er, 172 Yb, 165 Tm, 177 Lu, 225 Ac, 198 Au, 199 Au, and 227 Th.
• the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 67 Cu, 43 Sc, 44 Sc, 99m Tc, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 90 Y, 177 Lu, and 225 Ac. In some embodiments, the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, 155 Tb, 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• the optionally coordinated radiometal M is for use in diagnostic imaging and is selected from 61 Cu, 62 Cu, 64 Cu, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 52 Mn, 89 Zr, 99m Tc, 111 In, 152 Tb, and 155 Tb. In some embodiments, the optionally coordinated radiometal M is for use in diagnostic imaging and is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, and 155 Tb.
• the optionally coordinated radiometal M is for use in radionuclide therapy and is selected from 67 Cu, 90 Y, 105 Rh, 117m Sn, 149 Pm, 153 Sm, 161 Tb, 149 Tb, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 225 Ac, and 213 Bi.
• the optionally coordinated radiometal M is for use in radionuclide therapy and is selected from 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• the compounds of the disclosure can be prepared by methods known in the field of synthetic chemistry and as described in the examples of the present disclosure.
• the disclosure provides pharmaceutical compositions comprising a diagnostically effective amount or a therapeutically effective amount of at least one compound of the disclosure and a pharmaceutically acceptable carrier therefor.
• a pharmaceutically acceptable carrier which is present in an appropriate dosage, includes solvents, dispersion media, antibacterial and antifungal agents, isotonic agents, and the like, which are physiologically acceptable. The use of such media and agents are well- known in the art.
• the disclosure provides compounds and/or pharmaceutical compositions of the disclosure (i) for use as a diagnostic imaging agent, (ii) for use in radionuclide therapy or (iii) for use as a theragnostic agent, e.g.
• the subject of the methods of the present disclosure is a mammal, such as an animal or a human.
• the subject is a human.
• the compounds and/or pharmaceutical compositions of the disclosure may be used for diagnostic imaging, i.e. for use in diagnostic imaging of a cell or population of cells expressing a folate-receptor in vitro or in vivo, i.e. for convenient and effective administration to a subject in need for diagnostic imaging.
• the compounds and/or pharmaceutical compositions of the disclosure may be used for diagnostic imaging of a cell or population of cells expressing a folate- receptor comprising the steps of administering at least one compound and/or pharmaceutical compositions of the disclosure in diagnostically effective amounts and obtaining diagnostic image of the cell or population of cells expressing a folate-receptor.
• the compounds and/or pharmaceutical compositions of the disclosure may be used as a diagnostic imaging agent for therapeutic planning and/or monitoring the effectiveness of an ongoing therapeutic treatment.
• the present disclosure provides a method for diagnostic imaging of a cell or population of cells expressing a FR, said method comprising the steps of administering at least one compound or composition of the present disclosure in a diagnostically effective amount, and obtaining a diagnostic image of said cell or population of cells.
• the present disclosure provides a method for in vitro detection of a cell, e.g. a tumor cell, expressing the folate receptor in a tissue sample, e.g. a tissue biopsy taken from a subject, which includes contacting said tissue sample with a compound or composition of the present disclosure in diagnostically effective amounts and for sufficient time and conditions to allow binding to occur and detecting such binding by imaging techniques, such as PET imaging.
• the present disclosure provides a method for diagnostic imaging or monitoring (e.g. cancer therapy) a subject comprising the steps of (i) administering to the subject at least one compound and/or pharmaceutical composition of the present disclosure in a diagnostically effective amount, and (ii) performing diagnostic imaging using PET by detecting a signal from said at least one compound and/or pharmaceutical composition of the present disclosure (to follow the course of cancer therapy and/or determine a therapeutically effective amount of at least one further compound of composition of the disclosure to be administered for treatment).
• the compounds and/or pharmaceutical compositions of the present disclosure may be used for radionuclide therapy, i.e. for convenient and effective administration to a subject in need for radionuclide therapy.
• the compounds and/or pharmaceutical compositions of the present disclosure may be used for radionuclide therapy comprising the steps of administering to the subject in need thereof at least one and/or pharmaceutical composition of the present disclosure in therapeutically effective amounts, localizing the at least one compound and/or pharmaceutical composition in a tissue to be treated, and subjecting the tissue to radiation to achieve the desired therapeutic effect.
• the present disclosure provides a method for radionuclide therapy comprising the steps of administering to a subject in need thereof at least one compound or pharmaceutical composition of the present disclosure in therapeutically effective amounts, and after localization of said at least one compound or pharmaceutical composition in the desired tissues, subjecting the tissues to radiation to achieve the desired therapeutic effect.
• the compounds and/or pharmaceutical compositions of the present disclosure may be used as theragnostic agents for theragnostic applications.
• the term "theragnostic” is derived from therapy and diagnostics and refers with regard to applications or agents, to the strategy of utilising the same radioactively labelled drug eventually containing a different radionuclide, for diagnostics and for therapy. This allows to take images of a disease with a compound of the disclosure coordinated to a radionuclide effective for diagnostic and treatment planning purposes. It is then possible to treat the disease by changing to a radionuclide effective for tumor treatment. This is the so called ‘treat what you see' principle.
• the same compound of the disclosure may be used as a theragnostic agent first as a diagnostic imaging agent (a “diagnostic compound of the disclosure”) in a diagnostically effective amount with a radionuclide effective for tumor localization, assessment, monitoring or therapy planning and second as a therapeutic agent (a “therapeutic compound of the disclosure”) in a therapeutically effective amount with a radionuclide effective for tumor treatment.
• a diagnostic imaging agent a “diagnostic compound of the disclosure”
• a therapeutic agent a “therapeutic compound of the disclosure” in a therapeutically effective amount with a radionuclide effective for tumor treatment.
• Combining both functions allows optimization of selectivity (i.e. biodistribution) and efficacy (i.e. effective dosage), e.g. by first localizing and monitoring a tumor or cancerous tissue using a diagnostic compound of the disclosure, and subsequently tailoring a suitable administration regimen (i.e.
• the compounds of the present disclosure may be used in the therapeutic planning and/or treatment and/or monitoring of a tumor (or cancerous tissue) by administering to a subject in need thereof (i) at least one compound or composition of the disclosure in a diagnostically effective amount (i.e. an amount effective to obtain a diagnostic image and/or for treatment planning, i.e. establishing a treatment regimen), and (ii) at least one further compound or composition of the disclosure in a therapeutically effective amount for tumor treatment (by subjecting the tumor (or cancerous tissue) to radiation) to achieve the desired therapeutic effect.
• the diagnostically effective amount is an amount effective for diagnostic imaging, i.e.
• the at least one compound or composition of the present disclosure in a diagnostically effective amount is a diagnostic compound coordinating a radiometal M for use in diagnostic imaging as defined herein.
• the at least one further compound or composition of the present disclosure in a therapeutically effective amount is a therapeutic compound coordinating a radiometal M for use in radionuclide therapy as defined herein.
• a “diagnostically effective amount” of a compound or composition of the present disclosure to be administered is an amount sufficient to produce a diagnostic image of a tumor, a cancerous tissue, an organ or other site of the subject and/or an amount sufficient to determine the therapeutically effective amount for a treatment.
• a diagnostically effective amount of a compound or composition of the present disclosure is administered to monitor tumor growth or size before, during and after radionuclide therapy, and allows planning, tailoring and adjusting the therapy during the course of a treatment. In theragnostic applications, obtained results of the administration of a diagnostically effective amount of a compound or composition of the present disclosure are used to calculate the therapeutically effective amount.
• a “therapeutically effective amount” of a compound or composition of the present disclosure to be administered is an amount sufficient to produce a desired radiotherapeutic effect. More specifically, a therapeutically effective amount is an amount of at least one of the compounds of the present disclosure sufficient to substantially improve, i.e. ameliorate, decrease or suppress, at least one symptom associated with the disease or condition, and/or to delay, hinder, or prevent the onset of the disease or condition.
• a diagnostic compound of the present disclosure is a compound of the present disclosure wherein the radiometal M is a diagnostic radionuclide permitting diagnosis of a tumor.
• a diagnostic radionuclide is selected from 61 Cu, 62 Cu, 64 Cu, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 52 Mn, 89 Zr, 99m Tc, 111 In, 152 Tb, and 155 Tb. In some embodiments, a diagnostic radionuclide is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, and 155 Tb. In some embodiments, a therapeutic compound of the present disclosure is a compound of the present disclosure wherein the radiometal M is a therapeutic radionuclide permitting treatment of a tumor.
• a therapeutic radionuclide is selected from 67 Cu, 90 Y, 105 Rh, 117m Sn, 149 Pm, 153 Sm, 161 Tb, 149 Tb, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 225 Ac, and 213 Bi.
• a therapeutic radionuclide is selected from 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac. It is understood that a diagnostic compound of the present disclosure and a therapeutic compound of the present disclosure are administered to a subject sequentially.
• the present disclosure provides a method for therapeutic planning of a treatment in a subject comprising the steps of (i) administering to a subject in need thereof at least one compound or composition of the present disclosure in diagnostically effective amounts to obtain a diagnostic image, and (ii) determining the therapeutically effective amount of at least one further compound or composition of the present disclosure to be administered for treatment.
• the present disclosure provides a method for a theragnostic application in a subject comprising the steps of (i) administering to a subject in need thereof at least one compound or composition of the present disclosure in diagnostically effective amounts to obtain a diagnostic image, and (ii) administering at least one further compound or composition of the present disclosure in therapeutically effective amounts for treatment (i.e.
• the diagnostic image obtained of the at least one compound or composition in the tissue to be treated is used to determine (or calculate) the therapeutically effective amount of the compound or composition of the present used in for treatment.
• the present disclosure provides a method for a theragnostic application, i.e.
• the at least one compound or composition of the present disclosure in a diagnostically effective amount is a diagnostic compound coordinating a radiometal M for use in diagnostic imaging as defined herein.
• the at least one further compound or composition of the present disclosure in a diagnostically effective amount is a therapeutic compound coordinating a radiometal M for use in radionuclide therapy as defined herein.
• the diagnostic image of the at least one compound or composition in the tissue to be treated is used to determine (e.g. calculate) the therapeutically effective amount of the at least one further compound or composition administered to obtain a therapeutic effect.
• An image of a cell or tissue expressing the FR, i.e. a tumor cell or tissue, labeled with one or more of the compounds or compositions of the present disclosure can be detected using a radiation detector, e.g. a ⁇ -radiation detector.
• a radiation detector e.g. a ⁇ -radiation detector.
• One such procedure utilizes scintigraphy.
• the unit dose to be administered has a radioactivity of about 0.1 MBq to about 10 4 MBq. In some embodiments, the unit dose to be administered for diagnostic imaging has a radioactivity of about 1 MBq to about 1'000 MBq, such as about 100 MBq to 600 MBq. In some embodiments, the unit dose to be administered for radionuclide therapy has a radioactivity of about 1'000 MBq to about 10 4 MBq, such as about 5'000 MBq to 8'000 MBq. For a solution to be injected a preferred unit dosage is from about 0.01 mL to about 10 mL. After e.g.
• intravenous administration imaging of the organ or tumor in vivo can take place, if desired, from within minutes to hours or even longer, after the radiolabeled reagent has been administered to a subject.
• the compounds and/or compositions of the present disclosure may be administered by an appropriate route such as parentally (for example, intravenously), intramuscularly or intraperitoneally or by any other suitable method.
• the compounds and/or compositions of this disclosure may be administered to a subject by bolus or slow infusion intravenous injection.
• the suitable forms for injection include sterile aqueous solutions or dispersions and sterile powders of the above mentioned compounds and/or compositions of the present disclosure.
• the compounds or pharmaceutical compositions are generally sterile.
• Sterilization can be accomplished by any art recognized technique, including but not limited to, sterile filtration, addition of antibacterial of antifungal agents, for example, paraben, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
• Samples can be collected by procedures known to the skilled person, e.g., by collecting a tissue biopsy or a body fluid, by aspirating for tracheal or pulmonary samples and the like.
• Tissue samples to be tested include any tissue suspected to contain a cell expressing a FR, such as tumor cells, epithelial cells, kidneys, gastrointestinal or the hepatobiliary system, and others.
• Samples can be sectioned, e.g., with a microtome, to facilitate microscopic examination and observation of bound complex. Samples can also be fixed with an appropriate fixative either before or after incubation with one of the compounds or compositions of the present disclosure to improve the histological quality of sample tissues.
• Time and conditions sufficient for binding of a complex of the present disclosure to a FR on the cell include standard tissue culture conditions, i.e. samples can be cultured in vitro and incubated with one of the compounds or compositions of the present disclosure in physiological media. Such conditions are well known to the skilled person. Alternatively, samples can be fixed and then incubated with a complex or composition of the present disclosure in an isotonic or physiological buffer.
• a typical amount of said complex of the present disclosure for in vitro detection of a tumor cell can range from about 1 ng/L to about 1'000 ⁇ g/L. In some embodiments, the amount is about 1 ⁇ g/l to about 100 ⁇ g/L.
• the compounds of the disclosure for use in in vitro diagnosis of a tumor cell and for use in in vivo applications are compounds of the disclosure wherein the radiometal chelator is coordinating a radiometal M moiety selected from 51 Cr, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 47 Sc, 167 Tm, 141 Ce, 111 In, 168 Yb, 175 Yb, 140 La, 89 Zr, 90 Y, 88 Y, 153 Sm, 166 Ho, 52 Mn, 165 Dy, 166 Dy, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 97 Ru, 103 Ru, 186 Re, 188 Re, 203 Pb, 211 Bi, 212 Bi, 213 Bi, 214 Bi, 105 Rh, 109 Pd, 212 Pb, 117m Sn, 149 Pm, 161 Tb, 149 Tb, 152 Tb, 155 Tb, 99m Tc, 165 Er, 169 Er, 172
• the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 67 Cu, 43 Sc, 44 Sc, 99m Tc, 149 Tb, 152 Tb, 155 Tb, 161 Tb, 90 Y, 177 Lu, and 225 Ac. In some embodiments, the optionally coordinated radiometal M is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, 155 Tb, 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• the optionally coordinated radiometal M for use in diagnostic imaging is selected from 61 Cu, 62 Cu, 64 Cu, 67 Ga, 68 Ga, 43 Sc, 44 Sc, 52 Mn, 89 Zr, 99m Tc, 111 In, 152 Tb, and 155 Tb. In some embodiments, the optionally coordinated radiometal M for use in diagnostic imaging is selected from 67 Ga, 68 Ga, 64 Cu, 43 Sc, 44 Sc, 99m Tc, 152 Tb, and 155 Tb.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 105 Rh, 117m Sn, 149 Pm, 153 Sm, 161 Tb, 149 Tb, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 225 Ac, and 213 Bi.
• the optionally coordinated radiometal M for use in radionuclide therapy is selected from 67 Cu, 90 Y, 161 Tb, 149 Tb, 177 Lu, and 225 Ac.
• samples can be incubated in the presence of a compound, then washed and counted in a standard scintillation counter.
• Alternative methods apply and are known to the skilled person. It is understood that the above methods of the disclosure may be performed in combination with any other methods of cancer diagnosis or therapy including methods using other already developed diagnostic and/or therapeutic agents and utilizing x-ray computed tomography (CT), magnetic resonance imaging (MRI), functional magnetic resonance imaging (fMRI), SPECT, optical imaging, and ultrasound.
• CT computed tomography
• MRI magnetic resonance imaging
• fMRI functional magnetic resonance imaging
• SPECT positron emission tomography
• optical imaging and ultrasound.
• diagnostic imaging or radionuclide therapy or theragnostic applications it may be convenient to prepare the compounds of the present disclosure at, or near, the site where they are to be used.
• a single or multi-vial kit containing all of the components needed to prepare compounds or compositions of this disclosure, other than the radiometal ion itself.
• a preferred single- vial kit of the present disclosure comprises a compound of the present disclosure, without the radiometal chelator being coordinated, and a source of a pharmaceutically acceptable reducing agent such as a stannous salt.
• the kit comprises optionally further additives, for example the kit is buffered with a pharmaceutically acceptable acid or base to adjust the pH to a desired value for complex formation.
• Such a single vial kit may optionally contain exchange ligands such as glucoheptonate, gluconate, mannitol, maleate, citric or tartaric acid and may also contain reaction modifiers, such as diethylenetriaminepentaacetic acid or ethylenediamine tetraacetic acid. Additional additives, such as solubilizers (for example a cyclodextrin), antioxidants (for example ascorbic acid) and/or fillers (for example, NaCl) may be employed to improve the radiochemical purity and stability of the final product, or to aid in the production of the kit.
• the radiometal will typically be added separately in the form of a solution.
• a preferred multi-vial kit of the present disclosure comprises, in one vial, the components, other than the radiometal itself, that is, an exchange ligand and a pharmaceutically acceptable reducing agent such as a stannous salt.
• a compound of the present disclosure, wherein the radiometal chelator, is contained in a second vial, as well as optional additives such as buffers appropriate to adjust the pH to its optimal value.
• the radiometal will be provided in form of a solution to be added. All components of a kit may be in liquid, frozen or dry form. In some embodiment, the kit components are provided in lyophilized form. All of the compounds, compositions and/or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure.
• Purification of the folate conjugates Purification of the final products was carried out using a Merck-Hitachi LaChrom HPLC system, equipped with a D-7000 interface, a L-7200 autosampler, a L-7400 UV detector, a L-7100 pump and a semi-preparative reversed-phase C18 column (5 ⁇ m, 10 ⁇ 150 mm, SunfireTM, Waters, Milford, US-MA).
• the products were eluted using variable gradient conditions in Milli-Q water/acetonitrile (MeCN)/TFA or Milli-Q water/MeCN/NH 4 HCO 3 systems at a flow rate of 4 mL/min.
• the fractions containing the pure product were collected in a round-bottom flask, frozen in liquid nitrogen and lyophilized for ⁇ 24 h. Characterization of the folate conjugates. The final products were characterized using high resolution MALDI-TOF-MS (Bruker UltraFlex II; Billarica, US-MA). Radiolabeling: The folate conjugates were dissolved in Milli-Q water containing 4–6% sodium L-ascorbate (0.5 M) to obtain a final folate concentration of 1 mM, which was frozen at -20°C as a stock solution.
• the conjugates were labeled with lutetium-177 (no-carrier- added, in 0.04 M HCl; ITM Medical Isotopes GmbH, Kunststoff, Germany) up to a molar activity of 50 MBq/nmol. Radiolabeling was performed after defrosting the stock solution in a mixture of sodium acetate (0.5 M) and HCl (0.05 M) and ascorbic acid (0.5M) at pH ca.4.5. The reaction mixture was incubated at 95 °C for 10–15 min.
• Radiolabeled conjugates were diluted in Milli-Q water containing penta-sodium diethylenetriamine pentaacetic acid (Na5-DTPA; 50 ⁇ M) and assessed using a Merck Hitachi LaChrom HPLC system consisting of a L-7100 pump, a D-7000 interface, a L-7200 autosampler, a radioactivity detector (LB 506 B, Berthold Technologies GmbH) and a reversed-phase C18 column (5 ⁇ m, 4.6 ⁇ 150 mm, Xterra TM , Waters, USA).
• Na5-DTPA penta-sodium diethylenetriamine pentaacetic acid
• Example 1 Solid-phase synthesis of 6R-RedFol-24 and 6S-RedFol-24 2-Chlorotrityl chloride (2-CTC) resin (0.1 mmol) was weighed into a filter-containing 5 mL- syringe and swelled in anhydrous DCM for 45 min.
• N ⁇ -fluorenylmethyloxycarbonyl-N ⁇ -(4- allyloxycarbonyl)-L-lysine (Fmoc-Lys(Alloc)-OH (0.12 mmol, 1.2 equiv)) was dissolved in dry DCM in the presence of diisopropylethylamine (DIPEA, 0.8 mmol, 8.0 equiv), added to the resin, and stirred overnight (o/n). Residual reactants were removed after each reaction step by washing the resin three times with dimethylformamide (DMF) or DCM depending on the utilized solvent.
• DIPEA diisopropylethylamine
• N ⁇ -1-(4,4-dimethyl-2,6- dioxocyclohex-1-ylidene)ethyl-N ⁇ -Fmoc-L-lysine (Dde-Lys(Fmoc)-OH, 0.4 mmol, 4.0 equiv) was activated for one minute with O-(benzotriazol-1-yl)-N,N,N',N'- tetramethyluronium-hexafluorophosphate (HBTU, 0.396 mmol, 3.96 equiv) in the presence of DIPEA (0.8 mmol, 8.0 equiv) in dry DMF, added to resin-immobilized compound 1, and reacted for 1 h.
• DIPEA 0.8 mmol, 8.0 equiv
• Compound 5 was obtained by coupling activated Fmoc-L-glutamic acid 5-tert-butyl ester (Fmoc-Glu-O(tert-butyl), 0.4 mmol, 4.0 equiv) for 1.5 h and removal of the Fmoc protecting group.10-formyl-5- methyl-(6S)-tetrahydropteroic acid (6S-5-MTHP) was activated, added to resin-immobilized compound 5, and agitated for 2 h. The resultant resin-immobilized compound 6 was washed with DMF, DCM, and diethyl ether (Et 2 O) and dried under reduced pressure. Cleavage from the resin and removal of the protecting groups were carried out as described below.
• the precipitate was dissolved in a mixture of MeCN and aqueous ammonium hydrogen solution (aq. NH4HCO3, 50 mM; 1:1, v/v) for purification using reversed-phase high performance liquid chromatography (RP-HPLC).
• RP-HPLC reversed-phase high performance liquid chromatography
• the synthesis of the reduced folate conjugates 6S-RedFol-24 was performed according to the procedure described for 6R-RedFol-24 except that instead of 10-formyl-5-methyl-(6S)- tetrahydropteroic acid (6S-5-MTHP) 10-formyl-5-methyl-(6R)-tetrahydropteroic acid (6R-5-MTHP) was conjugated to the resin-immobilized joint precursor under nitrogen atmosphere and exclusion of light.
• Example 4 PBS/n-Octanol Distribution Coefficient (LogD Values)
• the n-octanol/PBS distribution coefficients (logD values) were determined for each folate radioconjugate of the disclosure in order to assess their hydrophilic/hydrophobic properties.
• the logD values of the folate radioconjugates of the disclosure 50 MBq/nmol were determined as previously reported, after dilution in PBS pH 7.4 to obtain an activity concentration of 10 MBq/500 ⁇ L.
• a sample of each radiofolate (ca.0.5 MBq, 25 ⁇ L, 0.01 nmol) was added to a mixture of 1475 ⁇ L PBS pH 7.4 and 1500 ⁇ L n-octanol.
• the amount of serum albumin in mouse (MSA) and human (HSA) blood plasma was defined as 550 ⁇ M and 800 ⁇ M, respectively, based on measurements using a dry chemistry analyzer (DRI-CHEM 4000i, FUJIFILM, Japan).
• the folate radioconjugates of the disclosure 50 MBq/nmol, ca.300 kBq, 0.006 nmol in 15 ⁇ L were added to samples of mouse and human blood plasma (150 ⁇ L), followed by incubation of the samples at 37 °C for 30 min.
• the samples were loaded on Amicon centrifugal filters (cut-off of 10 kDa; Merck Millipore) followed by centrifugation (14,000 rcf, 30 min, 4 °C) to allow the separation of the plasma-bound from the plasma- unbound (free) fractions of each sample.
• the inserts of the filter devices were inverted and centrifuged at 200 rcf for 3 min to recover the protein-bound fraction.
• the activity of the protein-bound fraction as well as the activity in the filtrate and in the filter unit were measured separately in a gamma-counter (Perkin Elmer, Wallac Wizard 1480).
• the percentage of radioconjugate bound to mouse and human albumin, respectively, was set in relation to the total activity measured (Table 4).
• the KB cells were rinsed with PBS prior to the addition of FFRPMI medium without supplements (975 ⁇ L/well).
• the folate radioconjugates of the disclosure 50 MBq/nmol were added to each well in a volume of 25 ⁇ L (0.75 pmol, 38 kBq).
• KB tumor cells were co-incubated with excess folic acid (100 ⁇ M) to block the FRs on the cell surface.
• the KB tumor cells were rinsed three times with ice-cold PBS to determine total uptake of the folate radioconjugates of the disclosure.
• a stripping buffer solution of 0.1 M acetic acid and 0.15 M NaCl, aq., pH 3
• NaOH solution (1 M, aq., 1 mL
• the cell lysates were counted for activity in a gamma-counter (Perkin Elmer, Wallac Wizard 1480).
• the protein concentrations of each sample were determined using a Micro BCA Protein Assay kit (Pierce, Thermo Scientific) in order to standardize the measured activity to the average content of protein in a single well.
• Example 7 FR-binding Affinity (K D Values) The determination of the K D values was performed as previously reported (Deberle, L. M. et al; Bioconj Chem 2021, 32, p.1617). FR-positive IGROV-1 cells (human ovarian carcinoma cell line, kindly provided by Dr. Gerrit Janssen, Free University Medical Center Amsterdam, The Netherlands) were seeded in 48-well plates (2.5 ⁇ 10 5 cells/well) in 500 ⁇ L FFRPMI medium with supplements.
• the cells were incubated at 37 °C and 5% CO2 to allow cell adhesion overnight. The experiment was performed on ice using ice-cold medium and buffer solutions. After removal of the supernatant, the cells were rinsed once with PBS prior to the addition of FFRPMI medium without supplements (450 ⁇ L/well).
• the respective folate radioconjugate of the disclosure (20 MBq/nmol) was added to each well at variable concentrations (in the range of 0.1 to 500 nM, 50 ⁇ L per well). In order to determine the unspecific binding of the folate radioconjugates of the disclosure, half of the cell samples were co-incubated with excess folic acid (100 ⁇ M) to block FRs on the cell surface.
• the IGROV-1 tumor cells were incubated for 1 h at 4 °C on a shaker, followed by removal of the supernatants and rinsing the cells twice with PBS. After lysis of the cells with NaOH solution (1 M, aq., 500 ⁇ L), the samples were counted for activity in a ⁇ -counter (Wallac Wizard 1480, Perkin Elmer). The counts per minute (cpm) of the specific binding (determined by subtracting the cpm of the unspecific binding from the cpm of total binding), were plotted against the molar concentration of the added folate radioconjugates of the disclosure. The nonlinear regression analysis for determination of the K D value was performed using GraphPad Prism software (version 8).
• mice were subcutaneously inoculated with KB tumor cells (5 ⁇ 10 6 cells in 100 ⁇ L of PBS) on the shoulder.
• the mice were sacrificed and selected tissues and organs were collected, weighed and counted for activity using a gamma-counter (PerkinElmer Wallac Wizard 1480).
• Example 9 SPECT/CT Imaging Studies SPECT/CT imaging studies were performed to investigate the whole body distribution of folate radioconjugates of the disclosure in KB tumor-bearing mice ( Figures 7). SPECT/CT experiments were performed approximately 2 weeks after tumor cell inoculation when the tumor size reached a volume of ca.300 mm 3 . Mice were injected with the folate radioconjugates of the disclosure (25 MBq, 0.5 nmol, 100 ⁇ L, diluted in PBS containing 0.05% BSA) and scanned at 1 h, 4 h, 24 h and 48 h p.i.
• Imaging studies were performed using a four-head, multiplexing, multipinhole small-animal SPECT camera (NanoSPECT/CTTM, Mediso Medical Imaging Systems, Budapest, Hungary) as previously reported. Each head was outfitted with a tungsten-based aperture of nine 1.4 mm-diameter pinholes and a thickness of 10 mm. CT scans of ca.7.5 min duration were followed by SPECT scans of ca.40 min. The images were acquired using Nucline Software (version 1.02, Mediso Ltd., Budapest, Hungary). The real-time CT reconstruction used a cone-beam filtered backprojection.
• Figures 7 is showing SPECT/CT images as maximum intensity projections (MIPs) of KB tumor-bearing mice 1 h, 4 h, and 24 h after injection of the 177 Lu-folate radioconjugates (25 MBq; 0.5 nmol per mouse).
• MIPs maximum intensity projections

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• 2023
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  • 2023-01-30 KR KR1020247028652A patent/KR20240146013A/ko active Pending
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  • 2023-01-30 CN CN202380018495.8A patent/CN118647412A/zh active Pending
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