Radiopharmaceutical complexes targeting prostate specific membrane antigens and combinations thereof

Abstract

The present invention relates to tissue targeting compounds. In particular, the present invention relates to tissue targeting compounds comprising a monoclonal antibody or antigen binding fragment thereof having binding affinity for prostate specific membrane antigen (PSMA). Furthermore, the present invention relates to a combination, preferably a pharmaceutical combination comprising a tissue targeting compound of formula (I) and another agent. The combinations are useful in therapy, preferably in the treatment of hyperproliferative diseases such as cancer. # imgabs0 #

Description

Field of the Invention

[0001] The present invention relates to tissue targeting compounds. In particular, the present invention relates to tissue targeting compounds comprising a monoclonal antibody or an antigen binding fragment thereof having binding affinity for prostate specific membrane antigen (PSMA). In addition, the present invention relates to a combination, preferably a pharmaceutical combination comprising a tissue targeting compound of formula (I) and another agent. The tissue targeting compound and / or combination can be used for diagnosis and / or treatment, preferably for treatment, more preferably for treatment of hyperproliferative diseases, such as cancer.

[0002] In addition, the present invention relates to a method for preparing the tissue targeting compound and its use in diagnosing and / or treating diseases, preferably in treating diseases, especially treating hyperproliferative diseases, especially treating cancer. Background of the Invention

[0004] In mammalian subjects, specific cell killing may be necessary for the successful treatment of various diseases. A common example of this is in the treatment of malignant diseases such as sarcomas and carcinomas. However, the selective elimination of certain cell types may also play a key role in the treatment of other diseases, particularly proliferative and neoplastic diseases.

[0005] Currently, the most common methods of selective treatment are surgery and external beam irradiation. However, targeted radionuclide therapy is a promising and developing field that has the potential to deliver highly cytotoxic radiation specifically to cell types associated with the disease. The most common forms of radiopharmaceuticals currently authorized for use in humans use beta-emitting and / or gamma-emitting radionuclides. However, there is some interest in using alpha-emitting radionuclides in therapy because alpha-emitting radionuclides have more effective cell killing.

[0006] The radiation range of commonly used alpha emitters in physiological environments is generally less than 100 microns, equivalent to only a few cell diameters. This makes these sources well suited for treating tumors, including micrometastases, because they have the range to reach neighboring cells within the tumor, but if they have good targeting, little of the radiation energy will pass through the target cell. Therefore, it is not necessary to target every cell, but damage to surrounding healthy tissue can be minimized (see Feinendegen et al., Radiat Res 148:195-201 (1997)). In contrast, beta particles have a range of more than 1 mm in water (see Wilbur, Antibody Immunocon. Radiopharm. 4:85-96 (1991)).

[0007] Compared to the energy carried by beta particles, gamma rays, and X-rays, the energy of alpha particle radiation is high, typically 5-8 MeV, or 5 to 10 times the energy of beta particles and more than 20 times the energy of gamma rays. Therefore, this deposition of a large amount of energy over a very short distance gives alpha radiation a particularly high linear energy transfer (LET), a high relative biological efficacy (RBE), and a low oxygen enhancement ratio (OER) compared to gamma and beta radiation (see Hall, "Radiobiology for the radiologist", Fifth Edition, Lippincott Williams & Wilkins, Philadelphia PA, USA, 2000). This explains the special cytotoxicity of alpha-emitting radionuclides, and also places strict requirements on the biological targeting of such isotopes and the control and research level of the distribution of alpha-emitting radionuclides to avoid unacceptable side effects.

[0008] Currently, the application in radioimmunotherapy is mainly focused on 211At, 213Bi and 225Ac, which have been explored in clinical immunotherapy trials. However, although the use of macrocyclic complexes of radionuclides for targeted radiotherapy has been around for some time, the compounds currently used generally adopt the macrocyclic chelator DOTA (2,2′,2″,2″′-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid), which lacks stability and causes the radionuclide to dissociate from the chelating macrocycle, resulting in reduced selectivity and activity towards target tissues and increased toxicity towards non-target tissues.

[0009] For example, in preclinical studies, a one-step radiolabeling procedure using DOTA antibody chelator conjugate (ACC) was reported with a chelator to antigen (CAR) ratio of up to 10 (William F. Maguire, Michael R. McDevitt, Peter M. Smith-Jones, David A. Scheinberg. "Efficient one-step radiolabeling of monoclonal antibodies to high specific activity with actinium-225 for alpha-particle radioimmunotherapy of cancer", J. Nucl. Med. 2014; 55 (9): 1492–1498). However, after labeling for 2 hours at 37°C, quantitative labeling was not achieved, and a subsequent purification step was required to remove unlabeled Ac-225. This is in contrast to the tissue-targeted compounds of the present invention, in which Ac-225 can be quantitatively chelated after one hour at room temperature, thereby forming a complex with a significantly lower CAR ratio (<1).

[0010] Although the use of larger macrocyclic compounds can make the complexation of larger-sized radionuclides more stable, there is a need to develop targeted compounds with improved stability, selectivity and efficacy, especially in the field of hyperproliferative diseases such as cancer. For some types of cancer, especially prostate cancer, patients initially respond to treatment but eventually develop resistance (Swami U, McFarland TR, Nussenzveig R, Agarwal N. Advanced Prostate Cancer: Treatment Advances and Future Directions. Trends Cancer 2020; 6(8): 702-15 doi10.1016 / j.trecan.2020.04.010). Therefore, there is a need for new drugs that can selectively target specific cells and cell types in malignant diseases, which are very effective and can reduce, mitigate and / or avoid the prevalence of resistance. In particular, there is a need to provide new therapeutic agents for the treatment of cancer, especially prostate cancer, which is the most common cancer in men (Mattiuzzi C, Lippi G. Current Cancer Epidemiology. J Epidemiol Glob Health 2019; 9(4): 217-22 doi10.2991 / jegh.k.191008.001). The prostate-specific membrane antigen (PSMA) encoded by the FOLH1 (folate hydrolase 1) gene is highly expressed in prostate cancer cells, making it a suitable target for radiotherapy.

[0011] Surprisingly, it was found that the tissue targeting compounds of formula (I) of the present invention and the combination of tissue targeting compounds of formula (I) and other agents exhibited favorable properties in terms of stability, activity, efficacy and selectivity. Specifically, the tissue targeting compounds of formula (I) of the present invention exhibited higher immune response fraction (IRF) values ​​at low chelator to antigen (CAR) ratios and high monomer purity, resulting in higher scores of radionuclide labeled compounds with affinity for PSMA. In addition, the tissue targeting compounds of formula (I) of the present invention exhibited good biodistribution and low accumulation in the liver in vivo, which is a sign of reduced dissociation of the radionuclide from the chelating part and thus increased stability. In contrast to the examples previously reported in the literature, the tissue targeting compounds of formula (I) of the present invention can be labeled with radionuclides under very mild conditions and at lower temperatures (i.e., room temperature), thereby reducing antibody denaturation and the amount of non-PSMA binding parts in the final product. Therefore, the tissue targeting compounds of formula (I) of the present invention show a better ratio of tumor to liver, so that diseases can be treated more effectively and selectively, reducing damage to non-target tissues while maintaining high efficacy against cancer cells in target tissues. When used in combination with one or more other agents, a greater therapeutic effect may be achieved than when either compound is used alone.

[0012] In the field of radiopharmaceuticals, there is an increasing need to provide radionuclide-containing compounds and / or combinations comprising radionuclide-containing compounds that are stable for a sufficiently long time to allow them to be delivered to the administering physician and patient without suffering radiopharmaceutical degradation, for example, by dissociation of the radionuclide from the chelating agent. From a logistical point of view, increasing the stability of the radiopharmaceutical in the radionuclide-containing compound and / or combination comprising a radionuclide-containing compound is particularly desirable as this will allow the radiopharmaceutical-containing drug product to be reliably delivered between countries and / or continents. --+ It is desirable to obtain radionuclide-containing compounds and / or combinations comprising radionuclide-containing compounds that are stable for at least 48 hours, preferably for 96 hours, and / or have a monomer content of at least 85% for at least 48 hours, preferably at least 90% for at least 48 hours. The present invention solves these problems by providing tissue-targeted compounds of formula (I) and combinations comprising tissue-targeted compounds of formula (I) and other agents.

[0013] In particular, a combination comprising a tissue targeting compound of formula (I) and another agent of the present invention will have the following effects:

[0014] (1) produce greater efficacy in reducing tumor growth or even eliminating tumors than either agent alone,

[0015] (2) providing for administration of smaller amounts of the administered targeted compound and / or chemotherapeutic drug,

[0016] (3) provide patients with chemotherapy that is well tolerated and has fewer adverse pharmacologic complications than single-agent chemotherapy and certain other combination therapies,

[0017] (4) provide for the treatment of a wider variety of cancer types in mammals (particularly humans),

[0018] (5) provide higher response rates among patients receiving treatment,

[0019] (6) Providing longer survival time to treated patients compared to standard chemotherapy,

[0020] (7) provide a longer time until tumor progression, and / or

[0021] (8) Compared with other cancer drug combinations known to produce inhibitory effects, the efficacy and tolerability results are at least as good as those of the drugs used alone. Summary of the invention

[0022] The present invention relates to a tissue targeting compound of formula (I)

[0023]

[0024] wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for prostate-specific membrane antigen (PSMA).

[0025] definition

[0026] The term "comprising" as used in this specification includes "consisting of".

[0027] If any provision in this document is referred to as "as provided herein", it means that it may be referred to anywhere in this document.

[0028] In the context of the present invention, "tissue targeting" is used herein to indicate that the substance in question (i.e., a tissue targeting compound, a tissue targeting actinium complex, and / or a tissue targeting moiety, particularly when it is in the form of a tissue targeting complex as described herein) is used to preferentially localize itself (particularly localize any conjugated actinium complex) to at least one tissue site where its presence (e.g., to transmit radioactive decay) is desired. Thus, the tissue targeting compound, complex, group or moiety, upon administration to a subject, is used to provide greater localization to at least one desired site in the subject's body, as compared to the concentration of an equivalent complex without the targeting moiety. In the present case, the targeting moiety will preferentially bind to a cell surface receptor associated with a cancer cell or other receptor associated with the tumor microenvironment.

[0029] In the context of the present invention, the term "PSMA" refers to the prostate specific membrane antigen encoded by the FOLH1 (folate hydrolase 1) gene (Gene ID: 2346).

[0030] In the context of the present invention, the terms TLX592 and J592 are used interchangeably and refer to the same monoclonal antibody, namely a monoclonal antibody having binding affinity to the antigen PSMA (anti-PSMA antibody) developed by Telix Pharmaceuticals, which has been described in, for example, WO2021 / 000018.

[0031] TLX592 (or J592) comprises the heavy chain sequence according to SEQ ID NO. 19 and the light chain sequence according to SEQ ID NO. 20.

[0032] Further, TLX592 (or J592) comprises at least three CDR heavy chain sequences as shown in SEQ ID NO.21, SEQ ID NO.22 and SEQ ID NO.23 and three CDR light chain sequences as shown in SEQ ID NO.24, SEQ ID NO.25 and SEQ ID NO.26:

[0033] TLX592 CDR heavy chain sequence

[0034] SEQ ID NO.21: EYTIH

[0035] SEQ ID NO.22:N INPNNGGTTY NQKFED

[0036] SEQ ID NO.23: GW NFDY

[0037] TLX592 CDR light chain sequence

[0038] SEQ ID NO.24: KASQDVG TAVD

[0039] SEQ ID NO.25: W ASTRHT

[0040] SEQ ID NO.26: QQ YNSYPLT

[0041] The compound of formula (I) of the present invention comprises a chelating moiety comprising a complexed actinium atom (Ac). In the context of the present invention, the term "Ac" means an ion of at least one actinium isotope emitting alpha rays. Preferably, the actinium isotope emitting alpha rays is 225Ac. Preferably, the ion of at least one actinium isotope emitting alpha rays is a chelating moiety having three positive charges. 225 Ac, that is 225 Ac3+ Therefore, in the context of the present invention, the term "Ac" preferably but not exclusively means 225 Ac 3+ .

[0042] In the compounds of formula (I) of the present invention, the actinium (Ac) atom is shown as complexed with four oxygen atoms and two nitrogen atoms of the macrocycle and two carboxylate groups. Although this is the assumed complexation mode of the actinium atom, this description includes all possible and conceivable situations in which one or more bonds do not exist, such as the actinium atom is not bound to all heteroatoms or carboxylate groups of the macrocycle, etc.

[0043] In the context of the present invention, the term IRF refers to the immunoreactive fraction, i.e., the fraction of the labeled product (i.e., the compound of formula (I) of the present invention) that is capable of binding to the target (i.e., PSMA). The Lindmo assay (Lindmo T., et al. (1984) "Determination of the immunoreactive fraction of radiolabeled monoclonalantibodies by linear extrapolation to binding at infinite antigen excess." J. Immunol. Methods. 72, 77-89) is the most commonly used method for evaluating the immunoreactive fraction.

[0044] In the context of the present invention, the term CAR refers to the ratio of chelator to antigen, which is a measure of the specific activity of a radiolabeled compound (eg, a compound of formula (I) of the present invention).

[0045] The terms "polypeptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. These terms apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimetics of the corresponding natural amino acids, as well as natural amino acid polymers and non-natural amino acid polymers. Unless otherwise indicated, a particular polypeptide sequence also implicitly encompasses conservatively modified variants thereof.

[0046] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Similarly, nucleotides may be referred to by their commonly recognized single-letter codes.

[0047] The term "antibody" as used herein means an immunoglobulin molecule, which includes but is not limited to full-length antibodies and monovalent antibodies. A "full-length antibody" is preferably composed of four polypeptide chains, i.e., two heavy chains (H) and two light chains (L), which are usually interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region may include, for example, three domains CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain (CL). The VH and VL regions can be further subdivided into highly variable regions, called complementary determining regions (CDRs), interspersed with more conservative regions, called framework regions (FRs). Each VH and VL is usually composed of three CDRs and up to four FRs, arranged from the amino terminus to the carboxyl terminus, for example in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. As used herein, a "monovalent antibody" is preferably composed of three polypeptide chains, i.e., two heavy chains (H) and one light chain (L), which are usually interconnected by disulfide bonds. One heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region may comprise, for example, three domains CH1, CH2, and CH3. The other heavy chain consists only of a heavy chain constant region. The light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region consists of one domain (CL). The VH and VL regions can be further subdivided into highly variable regions, called complementarity determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). Each VH and VL is usually composed of three CDRs and up to four FRs, arranged from the amino terminus to the carboxyl terminus, for example, in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0048] As used herein, the term "complementarity determining region" (CDR; e.g., CDR1, CDR2, and CDR3) refers to the amino acid residues of an antibody variable domain whose presence is essential for antigen binding. Each variable domain typically has three CDR regions, designated CDR1, CDR2, and CDR3. Each complementarity determining region can comprise amino acid residues from a "complementarity determining region" as defined by Kabat (e.g., residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable domain and residues 31-35 (H1), 50-65 (H2), and 95-102 (H3) in the heavy chain variable domain; (Kabat et al., Sequences of Proteins of Immunolological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD. (1991)) and / or those residues from a "hypervariable loop" (e.g., residues 26-32 (L1), 50-52 (L2), and 91-96 (L3) in the light chain variable domain and residues 26-32 (H1), 53-55 (H2), and 96-101 (H3) in the heavy chain variable domain (Chothia and Lesk; J Mol. Biol 196:901-917 (1987). In certain cases, the complementarity determining regions may include amino acids from CDR regions and hypervariable loops according to the Kabat definition.

[0049] Depending on the amino acid sequence of the constant region of the heavy chain, intact antibodies can be assigned to different "classes". There are five main classes of intact antibodies: IgA, IgD, IgE, IgG and IgM, several of which can be further divided into "subclasses" (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2. The preferred immunoglobulin class used in the present invention is IgG.

[0050] The heavy chain constant regions corresponding to different classes of antibodies are called [α], [δ], [ε], [γ] and [μ], respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known. The antibodies used herein are conventionally known antibodies and their functional fragments.

[0051] "Functional fragments" or "antigen-binding antibody fragments" of antibodies / immunoglobulins are defined herein as fragments of antibodies / immunoglobulins that retain the antigen-binding region (e.g., the variable region of IgG). The "antigen-binding region" of an antibody is generally located in one or more hypervariable regions of the antibody, such as CDR1, -2, and / or -3 regions; however, variable "framework" regions may also play an important role in antigen binding, for example, by providing a scaffold for the CDRs.

[0052] "Functional fragments", "antigen-binding antibody fragments" or "antibody fragments" of the present invention include, but are not limited to, Fab, Fab', Fab'-SH, F(ab')2 and Fv fragments; diabodies; single domain antibodies (DAbs), linear antibodies; single chain antibody molecules (scFv); and multispecific antibodies formed from antibody fragments, such as bispecific and trispecific antibodies (CAK Borrebaeck, editor (1995) Antibody Engineering (Breakthroughs in Molecular Biology), Oxford University Press; R. Kontermann & S. Duebel, editors (2001) Antibody Engineering (Springer Laboratory Manual), Springer Verlag). Antibodies other than "multispecific" or "multifunctional" antibodies are understood to have the same binding sites at each site. F(ab')2 or Fab can be designed to minimize or completely eliminate the intermolecular disulfide bond interactions that occur between the CH1 and CL domains.

[0053] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain comprising at least a portion of a constant region. The term includes native sequence Fc regions and variant Fc regions. In one embodiment, the human IgG heavy chain Fc region extends from Cys226 or from Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) in the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is performed according to the EU numbering system (also referred to as the EU index), as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0054] Variants of antibodies or antigen-binding antibody fragments contemplated by the present invention are molecules in which the binding activity of the antibody or antigen-binding antibody fragment is retained.

[0055] “Binding proteins” considered by the present invention are, for example, antibody mimetics, such as Affibodies, Adnectins, Anticalins, DARPins, Avimers, Nanobodies (reviewed by Gebauer M. et al., Curr. Opinion in Chem. Biol. 2009; 13: 245-255; Nuttall SD et al., Curr. Opinion in Pharmacology 2008; 8: 608-617).

[0056] Herein, a "human" antibody or antigen-binding fragment thereof is defined as an antibody or antigen-binding fragment thereof that is non-chimeric (e.g., non-"humanized") and is not derived from (in whole or in part) a non-human species. A human antibody or antigen-binding fragment thereof may be derived from a human or may be a synthetic human antibody. A "synthetic human antibody" is defined herein as an antibody having a sequence that is derived in whole or in part from a synthetic sequence based on analysis of known human antibody sequences. Computer design of human antibody sequences or fragments thereof can be achieved, for example, by analyzing a database of human antibody or antibody fragment sequences and designing polypeptide sequences using the data obtained therefrom. Another example of a human antibody or antigen-binding fragment thereof is an antibody encoded by a nucleic acid isolated from a library of antibody sequences of human origin (e.g., a library based on antibodies taken from natural sources of humans). Examples of human antibodies include Antibodies described by et al. in Nature Biotech. 2000, 18: 853-856.

[0057] A "humanized antibody" or a humanized antigen-binding fragment thereof is defined herein as: (i) an antibody derived from a non-human source (e.g., a transgenic mouse with a heterologous immune system) that is based on a human germline sequence; (ii) an antibody in which the amino acids of the framework region of a non-human antibody are partially exchanged for human amino acid sequences by genetic engineering or (iii) CDR-grafted, in which the CDRs of the variable domain are from a non-human source, while one or more frameworks of the variable domain are from a human source and the constant domains (if any) are from a human source.

[0058] A "chimeric antibody" or antigen-binding fragment thereof is defined herein as an antibody or antigen-binding fragment thereof in which the variable domains are derived from a non-human source and some or all of the constant domains are derived from a human source.

[0059] The term "monoclonal antibody" as used herein refers to an antibody obtained from a group of substantially homogeneous antibodies, i.e., the individual antibodies constituting the antibody group are identical except for a small amount of mutations (e.g., naturally occurring mutations) that may exist. Therefore, the term "monoclonal" means that the antibody is not a mixture of discrete antibodies. Compared with polyclonal antibody preparations that usually contain different antibodies for different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is directed to a single determinant on an antigen. In addition to specificity, the advantage of monoclonal antibody preparations is that they are generally not contaminated by other immunoglobulins. The term "monoclonal" should not be interpreted as requiring the generation of antibodies by any particular method. The term monoclonal antibody specifically includes chimeric antibodies, humanized antibodies, and human antibodies.

[0060] An "isolated" antibody is one that has been identified and separated from the cellular components in which it is expressed. Contaminating cellular components are substances that may interfere with the diagnostic or therapeutic use of the antibody and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes.

[0061] An "isolated" nucleic acid is one that has been identified and separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that normally contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0062] As used herein, an antibody "specifically binds", "is specific for / directed against", or "specifically recognizes" an antigen of interest, such as a tumor-associated polypeptide antigen target or an antigen-binding polypeptide target (e.g., an antigen-binding antibody), means that the antibody binds to the antigen target with sufficient affinity so that the antibody can be used as a therapeutic agent for targeting cells or tissues expressing the antigen, or binds to cells or tissues of the antigen-binding polypeptide target with sufficient affinity so that the antibody can be used as a reversal agent to neutralize the therapeutic activity of the antigen-binding polypeptide (e.g., an antigen-binding antibody), and does not significantly cross-react with other proteins, or does not significantly cross-react with proteins other than orthologs and variants (e.g., mutant forms, splice variants, or proteolytic truncated forms) of the above-mentioned targets. As used herein, the term "specifically recognizes" or "specifically binds to" or "is specific for / directed against" a particular polypeptide or an epitope on a particular polypeptide target can be, for example, a monovalent K+ binding affinity exhibited by the antibody or its antigen-binding fragment to the antigen. D Less than about 10 -4 M, or less than about 10 -5 M, or less than about 10 -6 M, or less than about 10 -7 M, or less than about 10 -8 M, or less than about 10 -9 M, or less than about 10 -10 M, or less than about 10-11 M, or less than about 10 -12 M or less. If the antibody can distinguish between an antigen and one or more reference antigens, the antibody "specifically binds", "specific for / for" or "specifically recognizes" the antigen. In the most general form, "specific binding", "specifically binds to", "specific for / for" or "specific recognition" refers to the ability of an antibody to distinguish between a target antigen and an unrelated antigen, for example, as determined according to one of the following methods. Such methods include, but are not limited to, surface plasmon resonance (SPR), Western blots, ELISA, RIA, ECL, IRMA tests, and peptide scanning. For example, a standard ELISA assay can be performed. Scoring can be performed by standard color development (e.g., secondary antibodies with horseradish peroxidase and tetramethylbenzidine with hydrogen peroxide). The reaction in certain wells is scored by optical density, for example, at 450nm. A typical background (=negative reaction) may be 0.1OD; a typical positive reaction may be 1OD. This means that the difference between positive / negative is greater than 5 times, 10 times, 50 times, and preferably greater than 100 times. Typically, binding specificity is determined not with a single reference antigen but with a panel of about three to five unrelated antigens, such as milk powder, BSA, transferrin, etc.

[0063] "Binding affinity" or "affinity" refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule and its binding partner. Unless otherwise indicated, "binding affinity" as used herein refers to intrinsic binding affinity, which reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The dissociation constant "K D "Affinity" is often used to describe the affinity between a molecule (e.g., an antibody) and its binding partner (e.g., an antigen), that is, how tightly a ligand binds to a specific protein. Ligand-protein affinity is affected by non-covalent intermolecular interactions between the two molecules. Affinity can be measured by common methods known in the art (including those described herein). In one embodiment, the "K affinity" according to the present invention is D ” or “K D The "value" is measured by using a surface plasmon resonance assay using a suitable device, but not limited to a Biacore instrument, such as Biacore T100, Biacore T200, Biacore2000, Biacore 4000, Biacore 3000 (GE Healthcare Biacore, Inc.) or a ProteOn XPR36 instrument (Bio-Rad Laboratories, Inc.).

[0064] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which secreted immunoglobulins bind to Fcγ receptors (FcγRs) on certain cytotoxic cells (e.g., NK cells, neutrophils, and macrophages), enabling these cytotoxic effector cells to specifically bind to antigenic target cells and subsequently kill the target cells, for example, with cytotoxins. In order to assess the ADCC activity of an antibody of interest, an in vitro ADCC assay, such as that described in U.S. Pat. Nos. 5,500,362 or 5,821,337 or U.S. Pat. No. 6,737,056 (Presta), can be performed. Useful effector cells for such assays include PBMCs and NK cells.

[0065] "Complement dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (CIq) to antibodies (of the appropriate subclass) that are bound to their cognate antigens. To assess complement activation, a CDC assay may be performed, e.g., as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996). Polypeptide variants having altered Fc region amino acid sequences (polypeptides having variant Fc regions) and increased or decreased CIq binding are described, e.g., in U.S. Pat. No. 6,194,551 Bl and WO 1999 / 51642.

[0066] "Percentage (%) of sequence identity" relative to a reference polynucleotide or polypeptide sequence is defined as the percentage of nucleic acid or amino acid residues in a candidate sequence that are identical to nucleic acid or amino acid residues in a reference polynucleotide or polypeptide sequence, respectively, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percentage of sequence identity. Conservative substitutions are not considered part of sequence identity. Preferably, gapless alignments are used. Alignments for determining percentage of amino acid sequence identity can be achieved in various ways within the skill of the art, for example, using publicly available computer software, such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithm required for achieving maximum alignment over the full length of the compared sequences.

[0067] Substantial sequence identity / similarity can be understood as having at least 80% sequence similarity / identity with the complete sequence, and / or having at least 90% sequence similarity / identity with a specific binding region (e.g., CDR region). Preferred sequence similarity or more preferred identity can be at least 92%, 95%, 97%, 98% or 99%. The "BestFit" program of the University of Wisconsin Genetics Computer Group Version 10 software package can be used to determine sequence similarity and / or identity. The program uses the local had algorithm of Smith and Waterman with default values ​​of: Gap creation penalty = 8, Gap extension penalty = 2, average match = 2.912, average mismatch = 2.003.

[0068] The terms "polynucleotide" or "nucleic acid" are used interchangeably herein to refer to a nucleotide chain of any length, and include DNA and RNA. The nucleotides may be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into the chain by DNA or RNA polymerase. The polynucleotide may include modified nucleotides, such as methylated nucleotides and their analogs.

[0069] "Sequence homology" refers to the percentage of amino acids that are identical or represent conservative amino acid substitutions.

[0070] In addition, the nucleic acid sequence encoding the variable region of the heavy chain and / or light chain can be converted into a nucleic acid sequence encoding, for example, a full-length antibody chain, a Fab fragment or a scFv. The VL- or VH-encoding DNA fragment can be operably connected (so that the amino acid sequence encoded by the two DNA fragments conforms to the reading frame) to another DNA fragment encoding, for example, an antibody constant region or a flexible linker. The sequences of human heavy and light chain constant regions are known in the art (see, for example, Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242), and the DNA fragments comprising these regions can be obtained by standard PCR amplification.

[0071] To generate a polynucleotide sequence encoding an scFv, the VH- and VL-encoding nucleic acids can be operably linked to another fragment encoding a flexible linker so that the VH and VL sequences can be expressed as a continuous single-chain protein, with the VL and VH regions connected by the flexible linker (see, e.g., Bird et al., (1988) Science 242:423-426; Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883; McCafferty et al., Nature (1990) 348:552-554).

[0072] To express the antibody, its antigen-binding fragment or variant thereof, standard recombinant DNA expression methods can be used (see, e.g., Goeddel; Gene Expression Technology. Methods in Enzymology 185, Academic Press, San Diego, Calif. (1990)). For example, the DNA encoding the desired polypeptide can be inserted into an expression vector and then transfected into a suitable host cell. Suitable host cells are prokaryotic and eukaryotic cells. Examples of prokaryotic host cells are, for example, bacteria, and examples of eukaryotic host cells are yeast, insects and insect cells, plants and plant cells, transgenic animals, or mammalian cells. In some embodiments, the DNA encoding the heavy chain and the light chain are inserted into separate vectors. In other embodiments, the DNA encoding the heavy chain and the light chain are inserted into the same vector. It should be understood that the design of the expression vector, including the selection of regulatory sequences, is affected by factors such as the selection of the host cell, the expression level of the desired protein, and whether the expression is constitutive or inducible.

[0073] By inserting the DNA sequence encoding the desired protein into the operable reading phase with a functional promoter together with suitable translation initiation and termination signals, useful expression vectors for bacterial purposes have been constructed. The vector will contain one or more phenotype selectable markers and replication origins to ensure the maintenance of the vector, and if necessary, amplification in the host is provided. Suitable prokaryotic hosts for conversion include but are not limited to various species in Escherichia coli (E. coli.), Bacillus subtilis (bacillus subtilis), Salmonella typhimurium (salmonella typhimurium) and Pseudomonas, Streptomyces and Staphylococcus.

[0074] Bacterial vectors can be, for example, based on phage, plasmid or phagemid. These vectors can contain selectable markers and bacterial origins of replication, which are derived from commercially available plasmids that usually contain the elements of the known cloning vector pBR322 (ATCC 37017). After transforming a suitable host strain and growing the host strain to a suitable cell density, the promoter selected is de-repressed / induced by a suitable method (e.g., temperature conversion or chemical induction), and the cells are cultured for an additional time. Usually, the cells are harvested by centrifugation, broken by physical or chemical methods, and the resulting crude extract is retained for further purification.

[0075] In bacterial systems, a number of expression vectors may be advantageously selected depending on the intended use of the protein being expressed. For example, when large quantities of such proteins are to be produced, for example, to generate antibodies or to screen peptide libraries, vectors that direct the expression of high-level fusion protein products that are easily purified may be desirable.

[0076] The antibodies or antigen-binding fragments thereof or variants thereof of the present invention include naturally purified products, products of chemical synthesis methods, and products produced by recombinant technology from prokaryotic hosts, such as Escherichia coli, Bacillus subtilis, Salmonella typhimurium, and various species within the genera Pseudomonas, Streptomyces, and Staphylococcus, preferably from Escherichia coli cells.

[0077] Preferred regulatory sequences for mammalian host cell expression include high-level viral elements that direct protein expression in mammalian cells, such as promoters and / or enhancers derived from cytomegalovirus (CMV) (e.g., CMV promoter / enhancer), Simian virus 40 (SV40) (e.g., SV40 promoter / enhancer), adenovirus (e.g., adenovirus major late promoter (AdMLP)), and polyoma virus. Expression of the antibody can be constitutive or regulated (e.g., by addition or removal of small molecule inducers such as tetracycline in combination with the Tet system). For further description of viral regulatory elements and their sequences, see, e.g., US Pat. No. 5,168,062 to Stinski, US Pat. No. 4,510,245 to Bell et al., and US Pat. No. 4,968,615 to Schaffner et al. Recombinant expression vectors may also include an origin of replication and a selectable marker (see, e.g., US Pat. Nos. 4,399,216, 4,634,665, and 5,179,017). Suitable selectable markers include genes that confer resistance to drugs such as G418, puromycin, hygromycin, blasticidin, zeocin / bleomycin or methotrexate on the host cell into which the vector has been introduced, or use of auxotrophic selectable markers such as glutamine synthetase (Bebbington et al., Biotechnology (NY). 1992 Feb; 10(2): 169-75). For example, the dihydrofolate reductase (DHFR) gene confers resistance to methotrexate, the neomycin resistance gene confers resistance to G418, the bsd gene from Aspergillus terreus confers resistance to blasticidin, puromycin N-acetyltransferase confers resistance to puromycin, the Sh ble gene product confers resistance to zeocin, and resistance to hygromycin is conferred by the E. coli hygromycin resistance gene (hyg or hph). Selectable markers such as DHFR or glutamine synthetase can also be used in amplification techniques in conjunction with MTX and MSX.

[0078] Transfection of the expression vector into host cells can be performed using standard techniques, such as electroporation, nucleofection, calcium phosphate precipitation, lipofection, polycation-based transfection such as polyethyleneimine (PEI)-based transfection and DEAE-dextran transfection.

[0079] Suitable mammalian host cells for expressing the antibodies, antigen-binding fragments thereof, or variants thereof provided herein include, but are not limited to, Chinese hamster ovary (CHO cells), such as CHO-K1, CHO-S, CHO-K1SV [including dhfr-CHO cells, described in Urlaub and Chasin, (1980) Proc. Natl. Acad. Sci. USA 77:4216-4220 and Urlaub et al., Cell. 1983 Jun; 33(2):405-12, together with a DHFR selective marker (e.g., as described in RJ Kaufman and PA Sharp (1982) Mol. Biol. 159:601-621); and other knockout cells exemplified in Fan et al., Biotechnol Bioeng. 2012 Apr; 109(4):1007-15], NS0 myeloma cells, COS cells, HEK293 cells, HKB11 cells, BHK21 cells, CAP cells, EB66 cells, and SP2 cells.

[0080] Expression can also be transient or semi-stable in expression systems such as HEK293, HEK293T, HEK293-EBNA, HEK293E, HEK293-6E, HEK293-Freestyle, HKB11, Expi293F, 293EBNALT75, CHO Freestyle, CHO-S, CHO-K1, CHO-K1SV, CHOEBNALT85, CHOS-XE, CHO-3E7 or CAP-T cells (e.g., Durocher et al., Nucleic Acids Res. 2002 Jan 15;30(2):E9).

[0081] In some embodiments, the expression vector is designed so that the expressed protein is secreted into the culture medium in which the host cells are grown. The antibody, antigen-binding fragment thereof, or variant thereof can be recovered from the culture medium using standard protein purification methods.

[0082] The antibodies or antigen-binding fragments or variants thereof of the present invention can be recovered and purified from recombinant cell cultures by known methods, including but not limited to ammonium sulfate or ethanol precipitation, acid extraction, protein A chromatography, protein G chromatography, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxyapatite chromatography, mixed mode chromatography, and lectin chromatography. High performance liquid chromatography ("HPLC") can also be used for purification. See, e.g., Colligan, Current Protocols in Immunology, or Current Protocols in Protein Science, John Wiley & Sons, NY, NY, (1997-2001), e.g., Chapters 1, 4, 6, 8, 9, 10, each of which is incorporated herein by reference in its entirety.

[0083] The antibodies or antigen-binding fragments thereof or variants thereof of the invention include naturally purified products, products of chemical synthesis procedures, and products produced by recombinant techniques from eukaryotic hosts, including, for example, yeast, higher plants, insects, and mammalian cells. Depending on the host used in the recombinant production process, the antibodies of the invention may be glycosylated, or may be non-glycosylated. Such methods are described in many standard laboratory manuals, such as Sambrook, supra, sections 17.37-17.42; Ausubel, supra, chapters 10, 12, 13, 16, 18, and 20.

[0084] In preferred embodiments, the antibody is purified (1) to greater than 95% by weight of the antibody as determined, for example, by the Lowry method, UV-Vis spectroscopy, or by SDS-capillary gel electrophoresis (e.g., on a Caliper LabChip GXII, GX90, or Biorad Bioanalyzer apparatus), and in further preferred embodiments, greater than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence, or (3) to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue or, preferably, silver stain. Isolated naturally occurring antibodies include antibodies in situ within recombinant cells since at least one component of the antibody's natural environment will not be present. Ordinarily, however, isolated antibodies will be prepared by at least one purification step.

[0085] In the context of the present invention, the term "Ac225-Macropa-Pelgifatamab" refers to the following compound:

[0086]

[0087] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which is bound to the rest of the molecule via the terminal amino group of the lysine residue of the monoclonal antibody or the antigen-binding fragment thereof.

[0088] In the context of the present invention, the term "Ac225-DOTA-Pelgifatamab" refers to the following compound:

[0089]

[0090] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which is bound to the rest of the molecule via the terminal amino group of the lysine residue of the monoclonal antibody or the antigen-binding fragment thereof.

[0091] The present invention includes a combination of a tissue-targeted compound of formula (I) as described herein and one or more other compounds (i.e., other agents) having biological activity, preferably against cancer, especially prostate cancer. These compounds are defined by their class, so that those skilled in the art can identify compounds belonging to this class.

[0092] In the context of the present invention, nonsteroidal antiandrogens refer to antiandrogens with nonsteroidal chemical structures, which are usually selective and complete or silent antagonists of androgen receptors. Examples include, but are not limited to, darolutamide, bicalutamide, enzalutamide, apalutamide, flutamide, nilutamide, topirutamide, pukelutamide or badeglutamide or any combination thereof. In the context of the present invention, nonsteroidal antiandrogens are preferably selected from darolutamide and enzalutamide, and more preferably nonsteroidal antiandrogens are darolutamide. In certain embodiments, nonsteroidal antiandrogens are enzalutamide. In certain embodiments, nonsteroidal antiandrogens are apalutamide. In further embodiments, nonsteroidal antiandrogens are bicalutamide, flutamide or nilutamide. In other further embodiments, nonsteroidal antiandrogens are ARV-766, EPI-7386, CC-94676, AC-0176, HP-518 or TAS-3681. Preferably, the nonsteroidal antiandrogen is darolutamide.

[0093] In the context of the present invention, steroidal antiandrogens refer to antiandrogens with a steroid chemical structure, which are generally antagonists of androgen receptors. Examples include, but are not limited to, cyproterone acetate, allylestradiol, chlormadinone acetate, delmadinone acetate, progesterone caproate, hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, osaterone acetate, osantolone acetate, and spironolactone. In the context of the present invention, steroidal antiandrogens are preferably cyproterone acetate.

[0094] In the context of the present invention, androgen synthesis inhibitor refers to an enzyme inhibitor that prevents androgen biosynthesis. Examples include, but are not limited to, ketoconazole, abiraterone, aminoglutethimide, goserelin and seviteronel. In the context of the present invention, androgen synthesis inhibitor is preferably abiraterone or goserelin.

[0095] In the context of the present invention, antigonadotropins refer to compounds that inhibit the activity and / or downstream effects of one or both of gonadotropins, follicle stimulating hormone (FSH) and luteinizing hormone (LH). Examples include, but are not limited to, Abarelix, Danazol, Gestrinone, Propionate, Cetrorelix, Degarelix, Alagol sodium, Ganirelix, Linzagolix and Relugolix. In the context of the present invention, antigonadotropins are preferably Degarelix or Relugolix.

[0096] In the context of the present invention, a PARP inhibitor refers to a compound that is a pharmacological inhibitor of the enzyme poly-ADP ribose polymerase (PARP). Examples include, but are not limited to, Olaparib, Rucaparib, Veliparib, Niraparib, Talazoparib, Pamiparib, CEP 9722, E7016 and Iniparib. In the context of the present invention, a PARP inhibitor is preferably Olaparib or Rucaparib.

[0097] In the context of the present invention, an ATR inhibitor refers to a compound that is a pharmacological inhibitor of the enzyme ATR serine / threonine protein kinase, also known as ataxia telangiectasia and Rad3-related protein or FRAP-related protein 1 (FRP1). Examples include, but are not limited to, Berzosertib, BAY1895344 (Elimusertib), AZD6738, M6620, AZ20 and VE 821. In the context of the present invention, a PARP inhibitor is preferably Berzosertib or BAY1895344.

[0098] In the context of the present invention, an ATM inhibitor refers to a compound that is a pharmacological inhibitor of the enzyme ATM serine / threonine kinase. Examples include, but are not limited to, AZD1390, KU-55933, KU-60019, KU-59403, CP-466722, AZ31 / AZ32, AZD0156.

[0099] In the context of the present invention, a DNA-PK inhibitor refers to a compound that is a pharmacological inhibitor of DNA-dependent protein kinase (DNA-PK). Examples thereof include, but are not limited to, AZD7648, Nedisertib, VX-984, CC-115, Samotolisib, and BAY-8400. In the context of the present invention, a DNA-PK inhibitor is preferably Nedisertib or BAY-8400.

[0100] In the context of the present invention, AKT inhibitor refers to a compound that is a pharmacological inhibitor of serine / threonine kinase AKT. Examples thereof include but are not limited to Ipatasertib, afuresertib, miransertib, capivasertib, uprosertib and MK2206. In the context of the present invention, AKT inhibitor is preferably Ipatasertib.

[0101] In the context of the present invention, a Pi3K inhibitor refers to a compound that is capable of inhibiting one or more phosphoinositide 3-kinase (PI3K) enzymes, which are part of the PI3K / AKT / mTOR pathway. Examples include, but are not limited to, Copanlisib, Buparlisib, Duvelisib, Idelalisib, Paxalisib, Zandelisib, Inavolisib, Duvelisib, Alpelisib, and Umbralisib. In the context of the present invention, a Pi3K inhibitor is preferably Copanlisib.

[0102] In the context of the present invention, a PSMA-targeted beta emitter refers to a compound containing a radionuclide capable of emitting beta radiation. Examples thereof include, but are not limited to, (177Lu)-vipivotide tetraxetan (Pluvicto), 177Lu-PSMA-I&T, and PNT2002.

[0103] In the context of the present invention, immune checkpoint inhibitors refer to compounds that can block proteins called checkpoints produced by certain types of immune system cells (e.g., T cells) and certain cancer cells. Checkpoints help prevent excessive immune responses and sometimes prevent T cells from killing cancer cells. Examples include, but are not limited to, Atezolizumab, Durvalumab, Avelumab, Nivolumab, Pembrolizumab, and Ipilimumab.

[0104] In the context of the present invention, an alpha emitter refers to a compound containing a radionuclide capable of undergoing radioactive decay, wherein the radionuclide emits an alpha particle (helium nucleus), thereby transforming or "decaying" into a different nucleus, whose mass number decreases by four and whose atomic number decreases by two. Examples include, but are not limited to, 223Ra, J591-225Ac, and PSMA-617-225Ac.

[0105] In the context of the present invention, vaccine refers to a compound suitable for cell-based cancer immunotherapy of prostate cancer. Examples include, but are not limited to, sipoleucel T.

[0106] In the context of the present invention, a chemotherapeutic agent refers to a compound suitable for use in prostate cancer chemotherapy. Examples include, but are not limited to, paclitaxel, docetaxel, ixabepilone, vinorelbine, nocodazole, vincristine, colchicine and eribulin. In the context of the present invention, a chemotherapeutic agent is preferably paclitaxel or docetaxel.

[0107] In the context of the present invention, external beam radiation therapy refers to radiation therapy in which an external source of ionizing radiation is directed at and / or applied to a specific part of a subject's body.

[0108] In the context of the present invention, the term "combination" refers not only to a dosage form containing all components (i.e., a tissue targeting compound of formula (I) and another agent, also referred to as a fixed-dose combination), and a combination package containing components separated from each other, but also to components administered simultaneously or sequentially, as long as they are used to prevent or treat the same disease.

[0109] The amount of active ingredient administered can vary widely depending on such factors as the specific compound and dosage unit employed, the mode and timing of administration, the period of treatment, the age, sex and general condition of the patient being treated, the nature and extent of the condition being treated, the rate of drug metabolism and excretion, potential drug combinations and drug-drug interactions. Summary of the invention

[0111] Compound

[0112] According to a first aspect, the present invention relates to a tissue targeting compound of formula (I)

[0113]

[0114] wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for prostate-specific membrane antigen (PSMA).

[0115] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0116]

[0117] wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for prostate-specific membrane antigen (PSMA), wherein [Ab] is bound to the remainder of the molecule via the terminal amino group of a lysine residue of the monoclonal antibody or the antigen-binding fragment thereof.

[0118] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0119]

[0120] wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for prostate-specific membrane antigen (PSMA) selected from Pelgifatamab, J591 and TLX592.

[0121] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0122]

[0123] wherein [Ab] is an anti-PSMA monoclonal antibody or an antigen-binding fragment thereof selected from Pelgifatamab, J591 and TLX592.

[0124] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0125]

[0126] wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for prostate-specific membrane antigen (PSMA) selected from Pelgifatamab and J591.

[0127] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0128]

[0129] wherein [Ab] is an anti-PSMA monoclonal antibody or an antigen-binding fragment thereof selected from Pelgifatamab and J591.

[0130] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0131]

[0132] Wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity to prostate-specific membrane antigen (PSMA) J591.

[0133] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0134]

[0135] wherein [Ab] is the anti-PSMA monoclonal antibody J591 or an antigen-binding fragment thereof.

[0136] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0137]

[0138] Wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof that has binding affinity for prostate-specific membrane antigen (PSMA) Pelgifatamab.

[0139] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0140]

[0141] Wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof that has binding affinity for prostate-specific membrane antigen (PSMA) Pelgifatamab, wherein Pelgifatamab or its antigen-binding fragment is bound to the rest of the molecule via the terminal amino group of a lysine residue of the monoclonal antibody or its antigen-binding fragment.

[0142] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0143]

[0144] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof.

[0145] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0146]

[0147] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, wherein Pelgifatamab or the antigen-binding fragment thereof is bound to the rest of the molecule via the terminal amino group of a lysine residue of the monoclonal antibody or the antigen-binding fragment thereof.

[0148] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0149]

[0150] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises a heavy chain nucleotide sequence having sequence similarity or identity to SEQ ID NO.1 and a light chain nucleotide sequence having sequence similarity or identity to SEQ ID NO.2.

[0151] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0152]

[0153] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises a heavy chain sequence according to SEQ ID NO.1 and a light chain sequence according to SEQ ID NO.2.

[0154] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0155]

[0156] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising a heavy chain sequence according to SEQ ID NO.1 and a light chain sequence according to SEQ ID NO.2, wherein Pelgifatamab or an antigen-binding fragment thereof is bound to the rest of the molecule via the terminal amino group of a lysine residue of the monoclonal antibody or an antigen-binding fragment thereof.

[0157] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0158]

[0159] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10.

[0160] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0161]

[0162] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10, wherein Pelgifatamab or an antigen-binding fragment thereof is bound to the rest of the molecule via the terminal amino group of the lysine residue of the monoclonal antibody or the antigen-binding fragment thereof.

[0163] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0164]

[0165] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises the heavy chain sequence according to SEQ ID NO.1:

[0166] SEQ ID NO. 1 - Heavy chain

[0167]

[0168] And the light chain sequence according to SEQ ID NO.2:

[0169] SEQ ID NO. 2-Light chain

[0170]

[0171] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0172]

[0173] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises a heavy chain nucleotide sequence having sequence similarity or identity to SEQ ID NO.3 and a light chain nucleotide sequence having sequence similarity or identity to SEQ ID NO.4.

[0174] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0175]

[0176] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises a heavy chain amino acid sequence according to SEQ ID NO.3 and a light chain amino acid sequence according to SEQ ID NO.4.

[0177] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0178]

[0179] Wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises the heavy chain amino acid sequence according to SEQ ID NO.3:

[0180] SEQ ID NO.3-Heavy chain

[0181]

[0182] And the light chain amino acid sequence according to SEQ ID NO.4:

[0183] SEQ ID NO.4-Light chain

[0184]

[0185] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0186]

[0187] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises Figure 1 Heavy chain nucleotide sequences and / or deduced amino acid sequences having similarity or identity to the sequences shown, and Figure 2 The light chain nucleotide sequences and / or deduced amino acid sequences to which the sequences shown have similarity or identity.

[0188] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0189]

[0190] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising Figure 1 The heavy chain nucleotide sequence and / or deduced amino acid sequence of the sequence shown, and according to Figure 2 The light chain nucleotide sequence and / or deduced amino acid sequence of the sequence shown.

[0191] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0192]

[0193] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising Figure 3 At least three CDR heavy chain sequences and three CDR light chain sequences of the sequences shown.

[0194] In a further embodiment, the present invention relates to a tissue targeting compound of formula (I)

[0195]

[0196] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising Figure 3 At least three CDR heavy chain sequences and three CDR light chain sequences of the sequences shown, wherein Pelgifatamab or its antigen-binding fragment is bound to the rest of the molecule through the terminal amino group of the lysine residue of the monoclonal antibody or its antigen-binding fragment.

[0197] The tissue targeting compound of formula (I) of the present invention comprises a chelating moiety capable of binding actinium. Preferably, the chelating moiety comprises an 18-membered chelating moiety. Preferably, the chelating moiety comprises the macrocyclic chelator N,N′-bis[(6-carboxyl-2-pyridyl)methyl]-4,13-diaza-18-crown-6 (macropa).

[0198] Macrocyclic chelators macropa have been described, for example, in Thiele et al., Angew. Chem. Int. Ed. 2017, 56 (46), 14712 and references therein. Macropa chelators and their derivatives are also described in WO2018 / 183906 and WO2020 / 106886 and references therein.

[0199] The actinium complexing chelating moiety is preferably terminated with an isothiocyanate or carboxylic acid moiety to facilitate its coupling to the anti-PSMA antibody or antigen-binding fragment thereof. When a macropa chelator is used, one of the pyridine rings of the macropa chelator is typically substituted so that it can be coupled to the anti-PSMA antibody or antigen-binding fragment thereof. This can be a simple functional group (e.g., carboxylic acid -COOH) or a more complex chemical structure, as long as it is capable of coupling to the anti-PSMA antibody or antigen-binding fragment thereof. In particular, the substituents on the pyridine ring of the macropa chelator are preferably terminated with an isothiocyanate or carboxylic acid moiety.

[0200] The anti-PSMA antibody or antigen-binding fragment thereof is preferably coupled to the rest of the molecule forming the tissue targeting compound of the present invention via a lysine residue of the antibody. Thus, the actinium complexing chelating moiety and the anti-PSMA antibody or antigen-binding fragment thereof may be coupled via an amide or thiourea moiety. When the preferred macrocyclic chelator macropa is used, the terminal isothiocyanate or carboxylic acid moiety is coupled to the terminal amino group of the lysine residue in the anti-PSMA antibody or antigen-binding fragment thereof, resulting in the formation of a thiourea or amide group, respectively. Thus, the preferred actinium complexing chelating moiety comprising the macropa chelator is attached to the anti-PSMA antibody or antigen-binding fragment thereof via a thiourea or amide bond derived from the terminal amino group of the lysine residue of the anti-PSMA antibody or antigen-binding fragment thereof.

[0201] Preferred antibodies or antigen-binding fragments thereof in the compounds of the present invention have binding affinity for PSMA (anti-PSMA antibodies). Prostate-specific membrane antigen (PSMA) is an enzyme encoded by the FOLH1 (folate hydrolase 1) gene in humans. Specific binding fragments such as Fab, Fab', F(ab')2 and single-chain specific binding antibodies are typical fragments. Specifically, the anti-PSMA antibody is preferably selected from Pelgifatamab, J591 and TLX592 or antigen-binding fragments thereof. Preferably, the anti-PSMA antibody is selected from Pelgifatamab and J591 or antigen-binding fragments thereof. More preferably, the anti-PSMA antibody is Pelgifatamab or an antigen-binding fragment thereof.

[0202] Pelgifatamab (CAS Registry No. 2414550-93-7, INN published in WHO Recommended List 86, WHO Drug Information, Vol. 35, No. 3, 2021) is a monoclonal antibody with binding affinity to the antigen PSMA (anti-PSMA antibody). Its preparation, isolation and purification have been described in, for example, US8114965 (Pelgifatamab = clone006).

[0203] Pelgifatamab comprises a heavy chain nucleotide sequence according to SEQ ID NO.1:

[0204] SEQ ID NO.1- Heavy chain

[0205]

[0206] And the light chain nucleotide sequence according to SEQ ID NO.2:

[0207] SEQ ID NO.2- Light chain

[0208]

[0209] Furthermore, Pelgifatamab comprises a heavy chain amino acid sequence according to SEQ ID NO.3:

[0210] SEQ ID NO.3- Heavy chain

[0211]

[0212] And the light chain amino acid sequence according to SEQ ID NO.4:

[0213] SEQ ID NO.4- Light chain

[0214]

[0215] Furthermore, Pelgifatamab comprises Figure 1 The heavy chain nucleotide sequence of the sequence shown and the Figure 2 Light chain nucleotide sequence of the sequence shown.

[0216] Furthermore, Pelgifatamab comprises Figure 1 The heavy chain amino acid sequence of the sequence shown and according to Figure 2 Light chain amino acid sequence of the sequence shown.

[0217] Further, Pelgifatamab comprises at least three CDR heavy chain sequences as shown in SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences as shown in SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10:

[0218] Pelgifatamab CDR heavy chain sequence

[0219] SEQ ID NO.5: RYGMH

[0220] SEQ ID NO.6:VIWYDGSNKYYADSVKG

[0221] SEQ ID NO.7: GGDFLYYYYYGMDV

[0222] Pelgifatamab CDR light chain sequence

[0223] SEQ ID NO.8: RASQGISNYLA

[0224] SEQ ID NO.9: EASTLQS

[0225] SEQ ID NO.10: QNYNSAPFT

[0226] Furthermore, Pelgifatamab comprises Figure 3 At least three CDR heavy chain sequences and three CDR light chain sequences of the sequences shown.

[0227] J591 is a monoclonal antibody with binding affinity to the antigen PSMA (anti-PSMA antibody). Its preparation, isolation and purification have been described in, for example, WO2002 / 098897 and EP2277542.

[0228] Furthermore, J591 comprises a heavy chain sequence according to SEQ ID NO.11 and a light chain sequence according to SEQ ID NO.12:

[0229] SEQ ID NO.11:

[0230]

[0231] SEQ ID NO.12:

[0232]

[0233] Further, J591 comprises at least three CDR heavy chain sequences according to SEQ ID NO.13, SEQ ID NO.14 and SEQ ID NO.15 and three CDR light chain sequences according to SEQ ID NO.16, SEQ ID NO.17 and SEQ ID NO.18:

[0234] J591 CDR heavy chain sequence

[0235] SEQ ID NO.13: YTIH

[0236] SEQ ID NO.14:N INPNNGGTTY NQKFED

[0237] SEQ ID NO.15: GW NFDY

[0238] J591 CDR light chain sequence

[0239] SEQ ID NO.16: KASQDVG TAVD

[0240] SEQ ID NO.17: W ASTRHT

[0241] SEQ ID NO.18: QQ YNSYPLT

[0242] TLX592 is a monoclonal antibody with binding affinity to the antigen PSMA (anti-PSMA antibody) developed by Telix Pharmaceuticals, described, for example, in WO2021 / 000018.

[0243] TLX592 comprises a heavy chain sequence according to SEQ ID NO. 19 and a light chain sequence according to SEQ ID NO. 20:

[0244] SEQ ID NO.19:

[0245]

[0246] SEQ ID NO.20:

[0247]

[0248] Further, TLX592 comprises at least three CDR heavy chain sequences according to SEQ ID NO.21, SEQ ID NO.22 and SEQ ID NO.23 and three CDR light chain sequences according to SEQ ID NO.24, SEQ ID NO.25 and SEQ ID NO.26:

[0249] TLX592 CDR heavy chain sequence

[0250] SEQ ID NO.21: EYTIH

[0251] SEQ ID NO.22:N INPNNGGTTY NQKFED

[0252] SEQ ID NO.23: GW NFDY

[0253] TLX592 CDR light chain sequence

[0254] SEQ ID NO.24: KASQDVG TAVD

[0255] SEQ ID NO.25: W ASTRHT

[0256] SEQ ID NO.26: QQ YNSYPLT

[0257] combination

[0258] In another aspect, the present invention relates to a combination, preferably a pharmaceutical combination comprising a tissue targeting compound of formula (I) as described above and another agent. Said combination is useful for treatment, preferably for the treatment of a hyperproliferative disease such as cancer.

[0259] A combination comprising an effective amount of a tissue targeting compound of formula (I) as described above and another agent achieves a higher therapeutic effect than when either compound is used alone.

[0260] The relative proportions of the compounds in the combination may also be selected based on their respective mechanisms of action and disease biology. The relative proportions of the compounds may vary widely.

[0261] Where appropriate, the release of one or more of the drugs in the combination may also be controlled to provide the desired therapeutic activity in a single dosage form, combination package, kit or separate independent dosage forms.

[0262] In general, the combinations of the present invention will help to:

[0263] (1) produce greater efficacy in reducing tumor growth or even eliminating tumors than either agent alone,

[0264] (2) providing for administration of smaller amounts of targeted compounds and / or chemotherapeutic drugs,

[0265] (3) provide patients with chemotherapy that is well tolerated and has fewer adverse pharmacologic complications than single-agent chemotherapy and certain other combination therapies,

[0266] (4) provide for the treatment of a wider variety of cancer types in mammals (particularly humans),

[0267] (5) provide a higher response rate among the patients treated,

[0268] (6) provide longer survival times in treated patients compared with standard chemotherapy,

[0269] (7) provide a longer time until tumor progression, and / or

[0270] (8) Compared with known situations where other cancer drug combinations produce antagonistic effects, the efficacy and tolerability results produced were at least as good as those of the drugs used alone.

[0271] The tissue targeting compounds of formula (I) are compounds of the invention as described herein.

[0272] The other agent is selected from a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, a PARP inhibitor, an ATR inhibitor, an ATM inhibitor, a DNA-PK inhibitor, an AKT inhibitor, a Pi3K inhibitor, a PSMA-targeted beta emitter, an immune checkpoint inhibitor, an alpha emitter, a vaccine, and a chemotherapeutic agent.

[0273] Preferably, the other agent is selected from nonsteroidal antiandrogens, steroidal antiandrogens, androgen synthesis inhibitors, antigonadotropins, PARP inhibitors, AKT inhibitors, Pi3K inhibitors, PSMA-targeted beta emitters, immune checkpoint inhibitors, alpha emitters, vaccines and chemotherapeutic agents.

[0274] More preferably, the other agent is selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, a PARP inhibitor, a PSMA-targeted beta emitter, an alpha emitter, a vaccine, and a chemotherapeutic agent.

[0275] More preferably, the other agent is selected from the group consisting of nonsteroidal antiandrogens, steroidal antiandrogens, androgen synthesis inhibitors, antigonadotropins, and PARP inhibitors.

[0276] Even more preferably, the other agent is selected from the group consisting of nonsteroidal antiandrogens, steroidal antiandrogens, androgen synthesis inhibitors and antigonadotropins.

[0277] Even more preferably, the other agent is a nonsteroidal antiandrogen.

[0278] If the other agent is a nonsteroidal antiandrogen, it is preferably selected from darolutamide, bicalutamide, enzalutamide, apalutamide, flutamide, nilutamide, topirutamide, proxalutamide or bavdegalutamide or any combination thereof. More preferably, the nonsteroidal antiandrogen is selected from darolutamide and enzalutamide, and most preferably, the nonsteroidal antiandrogen is darolutamide. In certain embodiments, the nonsteroidal antiandrogen is enzalutamide. In certain embodiments, the nonsteroidal antiandrogen is apalutamide. In further embodiments, the nonsteroidal antiandrogen is bicalutamide, flutamide or nilutamide. In other further embodiments, the nonsteroidal antiandrogen is ARV-766, EPI-7386, CC-94676, AC-0176, HP-518 or TAS-3681. Preferably, the nonsteroidal antiandrogen is darolutamide.

[0279] If the other agent is a steroidal antiandrogen, it is preferably selected from cyproterone acetate, allylestradiol, chlormadinone acetate, delmadinone acetate, progesterone caproate, hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, osaterol acetate, oxentolone and spironolactone. More preferably, the steroidal antiandrogen is cyproterone acetate.

[0280] If the other agent is an androgen synthesis inhibitor, it is preferably selected from ketoconazole, abiraterone, aminoglutethimide, goserelin and seviternaine. More preferably, the androgen synthesis inhibitor is abiraterone or goserelin.

[0281] If the other agent is an antigonadotropin, it is preferably selected from abarelix, danazol, gestrinone, paraben, cetrorelix, degarelix, alagroglucose sodium, ganirelix, linzagolix and relugolix. More preferably, the antigonadotropin is degarelix or relugolix.

[0282] If the other agent is a PARP inhibitor, it is preferably selected from olaparib, rucaparib, veliparib, niraparib, talazoparib, pamiparib, CEP 9722, E7016 and iniparib. More preferably, the PARP inhibitor is olaparib or rucaparib.

[0283] If the further agent is an ATR inhibitor, it is preferably selected from Berzosertib, BAY1895344, AZD6738, M6620, AZ20 and VE 821. More preferably, the PARP inhibitor is Berzosertib or BAY1895344 (Elimusertib).

[0284] If the further agent is an ATM inhibitor it is preferably selected from AZD1390, KU-55933, KU-60019, KU-59403, CP-466722, AZ31 / AZ32 and AZD0156.

[0285] If the other agent is a DNA-PK inhibitor, it is preferably selected from ZD7648, Nindectinib, VX-984, CC-115, Samotolisib and BAY-8400. More preferably, the DNA-PK inhibitor is Nindectinib or BAY-8400.

[0286] If the other agent is an AKT inhibitor, it is preferably selected from ipasatib, afloxetine, milansertib, capasitinib, eupsitine and MK2206. More preferably, the AKT inhibitor is ipasatib.

[0287] If the other agent is a Pi3K inhibitor, it is preferably selected from Copanlisib, Buparlisib, Duvilisib, Idranib, Paxalisib, Zandelisib, Inarlisib, Duvilisib, Apellisib and Erbulisib. More preferably, the Pi3K inhibitor is Copanlisib.

[0288] If the other agent is a PSMA-targeted beta emitter, it is preferably (177Lu)-vipivotidetetraxetan (Pluvicto), 177Lu-PSMA-I&T and PNT2002.

[0289] If the other agent is an immune checkpoint inhibitor, it is preferably selected from atezolizumab, durvalumab, avelumab, nivolumab, pembrolizumab and ipilimumab.

[0290] If the other agent is an alpha emitter, it is preferably selected from 223Ra, J591-225Ac and PSMA-617-225Ac.

[0291] If the other agent is a vaccine, it is preferably Sipoleucel T.

[0292] If the other agent is a chemotherapeutic agent, it is preferably selected from paclitaxel, docetaxel, ixabepilone, vinorelbine, nocodazole, vincristine, colchicine and eribulin. More preferably, the chemotherapeutic agent is paclitaxel or docetaxel.

[0293] Therefore, in one embodiment, the invention relates to a combination comprising

[0294] (i) Tissue-targeting compounds of formula (I)

[0295]

[0296] wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for prostate-specific membrane antigen (PSMA), and

[0297] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, a PARP inhibitor, an ATR inhibitor, an ATM inhibitor, a DNA-PK inhibitor, an AKT inhibitor, a Pi3K inhibitor, a PSMA-targeted beta emitter, an immune checkpoint inhibitor, an alpha emitter, a vaccine, and a chemotherapeutic agent.

[0298] In a further embodiment, the invention relates to a combination comprising

[0299] (i) Tissue-targeting compounds of formula (I)

[0300]

[0301] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, and

[0302] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, a PARP inhibitor, an ATR inhibitor, an ATM inhibitor, a DNA-PK inhibitor, an AKT inhibitor, a Pi3K inhibitor, a PSMA-targeted beta emitter, an immune checkpoint inhibitor, an alpha emitter, a vaccine, and a chemotherapeutic agent.

[0303] In a further embodiment, the invention relates to a combination comprising

[0304] (i) Tissue-targeting compounds of formula (I)

[0305]

[0306] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, wherein Pelgifatamab or the antigen-binding fragment thereof is bound to the rest of the molecule via the terminal amino group of a lysine residue of the monoclonal antibody or the antigen-binding fragment thereof, and

[0307] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, a PARP inhibitor, an ATR inhibitor, an ATM inhibitor, a DNA-PK inhibitor, an AKT inhibitor, a Pi3K inhibitor, a PSMA-targeted beta emitter, an immune checkpoint inhibitor, an alpha emitter, a vaccine, and a chemotherapeutic agent.

[0308] In a further embodiment, the invention relates to a combination comprising

[0309] (i) Tissue-targeting compounds of formula (I)

[0310]

[0311] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising a heavy chain sequence according to SEQ ID NO.1 and a light chain sequence according to SEQ ID NO.2, and

[0312] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, a PARP inhibitor, an ATR inhibitor, an ATM inhibitor, a DNA-PK inhibitor, an AKT inhibitor, a Pi3K inhibitor, a PSMA-targeted beta emitter, an immune checkpoint inhibitor, an alpha emitter, a vaccine, and a chemotherapeutic agent.

[0313] In a further embodiment, the invention relates to a combination comprising

[0314] (i) Tissue-targeting compounds of formula (I)

[0315]

[0316] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10, and

[0317] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, a PARP inhibitor, an ATR inhibitor, an ATM inhibitor, a DNA-PK inhibitor, an AKT inhibitor, a Pi3K inhibitor, a PSMA-targeted beta emitter, an immune checkpoint inhibitor, an alpha emitter, a vaccine, and a chemotherapeutic agent.

[0318] In a further embodiment, the invention relates to a combination comprising

[0319] (i) Tissue-targeting compounds of formula (I)

[0320]

[0321] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising a heavy chain amino acid sequence according to SEQ ID NO.3 and a light chain amino acid sequence according to SEQ ID NO.4, and

[0322] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, a PARP inhibitor, an ATR inhibitor, an ATM inhibitor, a DNA-PK inhibitor, an AKT inhibitor, a Pi3K inhibitor, a PSMA-targeted beta emitter, an immune checkpoint inhibitor, an alpha emitter, a vaccine, and a chemotherapeutic agent.

[0323] In a further embodiment, the invention relates to a combination comprising

[0324] (i) Tissue-targeting compounds of formula (I)

[0325]

[0326] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10, and

[0327] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, a PARP inhibitor, a PSMA-targeted beta emitter, an alpha emitter, a vaccine, and a chemotherapeutic agent.

[0328] In a further embodiment, the invention relates to a combination comprising

[0329] (i) Tissue-targeting compounds of formula (I)

[0330]

[0331] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10, and

[0332] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, and a PARP inhibitor.

[0333] In a further embodiment, the invention relates to a combination comprising

[0334] (i) Tissue-targeting compounds of formula (I)

[0335]

[0336] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10, and

[0337] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, and an antigonadotropin.

[0338] In a further embodiment, the invention relates to a combination comprising

[0339] (i) Tissue-targeting compounds of formula (I)

[0340]

[0341] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10, and

[0342] (ii) Nonsteroidal antiandrogens.

[0343] In a further embodiment, the invention relates to a combination comprising

[0344] (i) Tissue-targeting compounds of formula (I)

[0345]

[0346] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10, and

[0347] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, a PARP inhibitor, a PSMA-targeted beta emitter (177Lu)-vipivotide tetraxetan (Pluvicto), an alpha emitter, the vaccine Sipoleucel T, and a chemotherapeutic agent, wherein the nonsteroidal antiandrogen is selected from the group consisting of darolutamide, bicalutamide, enzalutamide, apalutamide, flutamide, nilutamide, and topiramide, the steroidal antiandrogen is selected from the group consisting of cyproterone acetate, allylestradiol, chlormadinone acetate, delmadinone acetate, progesterone caproate, hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, osetaxelone acetate, orsendolone, and spironolactone, and the androgen The gonadotropin synthesis inhibitor is selected from ketoconazole, abiraterone, aminoglutethimide, goserelin and svetnet, the antigonadotropin is selected from abarelix, danazol, gestrinone, paraben, cetrorelix, degarelix, alagrocon sodium, ganirelix, linzagoli and relugolix, the PARP inhibitor is selected from olaparib, rucaparib, veliparib, niraparib, talazoparib, pamiparib, CEP 9722, E7016 and iniparib, the alpha emitter is selected from 223Ra, J591-225Ac and PSMA-617-225Ac, and the chemotherapeutic agent is selected from paclitaxel, docetaxel, ixabepilone, vinorelbine, nocodazole, vincristine, colchicine and eribulin.

[0348] In a further embodiment, the invention relates to a combination comprising

[0349] (i) Tissue-targeting compounds of formula (I)

[0350]

[0351] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10, and

[0352] (ii) an agent selected from the group consisting of a nonsteroidal antiandrogen, a steroidal antiandrogen, an androgen synthesis inhibitor, an antigonadotropin, and a PARP inhibitor, wherein the nonsteroidal antiandrogen is selected from the group consisting of darolutamide, bicalutamide, enzalutamide, apalutamide, flutamide, nilutamide, and topiramide, and the steroidal antiandrogen is selected from the group consisting of cyproterone acetate, allylestradiol, chlormadinone acetate, delmadinone acetate, progesterone caproate, hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, acetonide The invention relates to an in vitro induced dysmenorrhea drug comprising: abarelix, danazol, gestrinone, paraben, cetrorelix, degarelix, alagrogol sodium, ganirelix, linzagoli and relugolix, and the PARP inhibitor is selected from olaparib, rucaparib, veliparib, niraparib, talazoparib, pamiparib, CEP 9722, E7016 and iniparib.

[0353] In a further embodiment, the invention relates to a combination comprising

[0354] (i) Tissue-targeting compounds of formula (I)

[0355]

[0356] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10, and

[0357] (ii) an agent selected from the group consisting of nonsteroidal antiandrogens, steroidal antiandrogens, androgen synthesis inhibitors, and antigonadotropins, wherein the nonsteroidal antiandrogen is selected from the group consisting of darolutamide, bicalutamide, enzalutamide, apalutamide, flutamide, nilutamide, and topiramide, the steroidal antiandrogen is selected from the group consisting of cyproterone acetate, allylestradiol, chlormadinone acetate, delmadinone acetate, progesterone caproate, hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, osatetron acetate, oxentolone, and spironolactone, the androgen synthesis inhibitor is selected from the group consisting of ketoconazole, abiraterone, aminoglutethimide, goserelin, and sivetenolone, and the antigonadotropin is selected from the group consisting of abarelix, danazol, gestrinone, parahydroxypropiophenone, cetrorelix, degarelix, elagrogol sodium, ganirelix, linzagoli, and relugolix.

[0358] In a further embodiment, the invention relates to a combination comprising

[0359] (i) Tissue-targeting compounds of formula (I)

[0360]

[0361] wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, comprising at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10, and

[0362] (ii) an agent selected from the group consisting of the nonsteroidal antiandrogen darolutamide, the steroidal antiandrogen cyproterone acetate, the androgen synthesis inhibitors abiraterone or goserelin, and the antigonadotropins degarelix or relugolix.

[0363] synthesis

[0364] In another aspect, the present invention relates to a method for preparing a tissue targeting compound of formula (I),

[0365]

[0366] Wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for PSMA, the method comprises

[0367] (i) coupling the chelating moiety of formula (II) to a monoclonal antibody or antigen-binding fragment thereof having binding affinity for PSMA, and

[0368]

[0369] (ii) contacting the resulting product with an aqueous solution comprising ions of at least one alpha-emitting actinium isotope.

[0370] In a further embodiment, the present invention relates to a method for preparing a tissue targeting compound of formula (I),

[0371]

[0372] Wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for PSMA, the method comprises

[0373] (i) coupling the chelating moiety of formula (II) to a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for PSMA via a lysine residue of the monoclonal antibody or the antigen-binding fragment thereof, and

[0374]

[0375] (ii) contacting the resulting product with an aqueous solution comprising ions of at least one alpha-emitting actinium isotope.

[0376] In a further embodiment, the present invention relates to a method for preparing a tissue targeting compound of formula (I),

[0377]

[0378] wherein [Ab] is an anti-PSMA antibody or an antigen-binding fragment thereof selected from Pelgifatamab, J591 and TLX592,

[0379] The method comprises

[0380] (i) coupling a chelating moiety of formula (II) to an anti-PSMA antibody or an antigen-binding fragment thereof selected from Pelgifatamab, J591 and TLX592 via a lysine residue of the anti-PSMA antibody or the antigen-binding fragment thereof, and

[0381]

[0382] (ii) contacting the resulting product with an aqueous solution comprising ions of at least one alpha-emitting actinium isotope.

[0383] In a further embodiment, the present invention relates to a method for preparing a tissue targeting compound of formula (I),

[0384]

[0385] wherein [Ab] is the anti-PSMA antibody Pelgifatamab or an antigen-binding fragment thereof,

[0386] The method comprises

[0387] (i) coupling the chelating moiety of formula (II) to the anti-PSMA antibody Pelgifatamab or its antigen-binding fragment via a lysine residue of the monoclonal antibody or its antigen-binding fragment, and

[0388]

[0389] (ii) contacting the resulting product with an aqueous solution comprising ions of at least one alpha-emitting actinium isotope.

[0390] In step (i), the chelating moiety of formula (II) is coupled to a monoclonal antibody or antigen-binding fragment thereof having binding affinity for PSMA.

[0391]

[0392] Preferably, the anti-PSMA antibody is selected from Pelgifatamab, J591 and TLX592 or an antigen-binding fragment thereof. More preferably, the anti-PSMA antibody is Pelgifatamab or an antigen-binding fragment thereof.

[0393] Preferably, the chelating moiety of formula (II) is coupled to the anti-PSMA antibody or antigen-binding fragment thereof through a lysine residue of the monoclonal antibody, thereby forming a thiourea moiety linking the chelating moiety to the anti-PSMA antibody or antigen-binding fragment thereof.

[0394] Preferably, the coupling in step (i) is carried out at a basic pH, ie a pH above 7. More preferably, the coupling in step (i) is carried out at a pH of 8 to 10, most preferably at a pH of 9.

[0395] After step (i), a tissue targeting chelator of formula (III) is generated. Therefore, in a further embodiment, the present invention relates to a tissue targeting chelator of formula (III),

[0396]

[0397] Wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof that has binding affinity for PSMA.

[0398] Preferably, the anti-PSMA antibody or antigen-binding fragment thereof is selected from Pelgifatamab, J591 and TLX592 or antigen-binding fragments thereof. More preferably, the anti-PSMA antibody is Pelgifatamab or an antigen-binding fragment thereof.

[0399] In step (ii), the product obtained by coupling in step (i) is contacted with an aqueous solution containing ions of at least one alpha-emitting actinium isotope. Preferably, the alpha-emitting actinium isotope is 225Ac. Preferably, the ions of at least one alpha-emitting actinium isotope are ions having three positive charges. 225 Ac, that is 225 Ac 3+ The present invention allows step (ii) to be performed at room temperature, i.e., at a temperature lower than the temperature required for preparing the corresponding DOTA conjugate (i.e., Ac225-DOTA-Pelgifatamab), because the preparation of the corresponding DOTA conjugate (i.e., Ac225-DOTA-Pelgifatamab) requires a higher temperature (60°C).

[0400] use

[0401] On the other hand, the tissue targeting compound of formula (I) of the present invention and / or a combination comprising a tissue targeting compound of formula (I) and another agent of the present invention can be used to treat a disease in a human or non-human animal subject. In particular, the tissue targeting compound of formula (I) of the present invention and / or a combination comprising a tissue targeting compound of formula (I) and another agent of the present invention can be used to treat proliferative and / or neoplastic diseases, such as cancer. It is known that many targets are associated with proliferative diseases and neoplastic diseases. These include certain receptors, cell surface proteins, transmembrane proteins, and proteins / peptides found in the extracellular matrix near the diseased cells. Examples of cell surface receptors and antigens that may be associated with neoplastic diseases include, but are not limited to, CD22, CD33, FGFR2 (CD332), PSMA, HER2, GPC3, and mesothelin. In a preferred embodiment, the tissue targeting compound of formula (I) of the present invention and / or a combination comprising a tissue targeting compound of formula (I) of the present invention comprises a tissue targeting moiety, and the tissue targeting moiety comprises a monoclonal antibody having specificity and / or binding affinity for PSMA. For example, this can be reflected by a binding affinity for cells expressing PSMA that is 50 times or more (preferably at least 100 times higher, more preferably at least 300 times higher) higher than the binding affinity for cells that do not express PSMA. It is believed that PSMA is expressed and / or overexpressed in cells with certain disease states (as shown herein), so PSMA-specific binding agents can be used to target the complex to such cells affected by the disease. Similarly, the tissue targeting portion can bind to a cell surface marker (e.g., a PSMA receptor) on a cell near a diseased cell. The expression of the PSMA cell surface marker on the surface of a diseased cell may be higher than that on the surface of a healthy cell, or the expression on the cell surface during growth or replication may be higher than that in a dormant period. In one embodiment, a PSMA-specific tissue targeting adhesive can be used in combination with another adhesive for a disease-specific cell surface marker to obtain a dual binding complex. As discussed herein, the tissue targeting adhesive for PSMA is typically a peptide or protein.

[0402] Various aspects of the invention described herein relate to the treatment of disease, particularly selective targeted therapy for diseased tissue, and to complexes, conjugates, medicaments, formulations, kits, etc. useful in such methods. In all aspects, the diseased tissue may be located at a single site in the body (e.g., in the case of a localized solid tumor), or may be located at multiple sites (e.g., in the case of arthritis where multiple joints are affected or in the case of distributed or metastatic cancer).

[0403] The targeted diseased tissue can be located at a soft tissue site, a calcified tissue site, or multiple sites, which can all be located in soft tissue, all be located in calcified tissue, or can include at least one soft tissue site and / or at least one calcified tissue site. In one embodiment, at least one soft tissue site is targeted. The targeted site and the origin site of the disease can be the same, but can also be different (e.g., the metastatic site is specifically targeted). When multiple sites are involved, this may include the original site, and may also be multiple secondary sites.

[0404] The term "soft tissue" as used herein refers to a tissue that does not have a "hard" mineralized matrix. Specifically, the soft tissue used herein can be any tissue that is not a bone tissue. Accordingly, the "soft tissue disease" used herein means a disease that occurs in the "soft tissue" used herein. The present invention is particularly suitable for treating cancer and "soft tissue diseases", and therefore includes cancers, sarcomas, myelomas, leukemias, lymphomas and mixed cancers that occur in any "soft" (i.e., non-mineralized) tissue, as well as other non-cancerous diseases of such tissues. Cancerous "soft tissue diseases" include solid tumors that occur in soft tissues as well as metastatic and micrometastatic tumors. In fact, soft tissue diseases may include primary solid tumors of soft tissues and at least one soft tissue metastatic tumor in the same patient. Alternatively, "soft tissue diseases" may consist only of primary tumors, or only of metastatic tumors, wherein the primary tumor is a bone disease. In all appropriate aspects of the present invention, blood tumors, especially lymphocyte tumor diseases, such as lymphomas and lymphocytic leukemias, including non-Hodgkin's lymphomas, B-cell tumors of B-cell lymphomas, are particularly suitable for treatment and / or targeting. Similarly, any tumor disease of the bone marrow, spinal (especially spinal cord) lymph nodes and / or blood cells is suitable for treatment and / or targeting in all appropriate aspects of the present invention.

[0405] Examples of some B cell diseases (e.g., tumors) that are suitable for treatment and / or targeting by appropriate aspects of the present invention include:

[0406] Chronic lymphocytic leukemia / small lymphocytic lymphoma, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma (eg, Waldenstrom's macroglobulinemia macroglobulinemia), splenic marginal zone lymphoma, plasma cell neoplasms (e.g., plasma cell myeloma, plasmacytoma, monoclonal immunoglobulin deposition disease, heavy chain disease), extranodal marginal zone B-cell lymphoma (MALT lymphoma), nodal marginal zone B-cell lymphoma (NMZL), follicular lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, mediastinal (thymic) large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, and Burkitt lymphoma / leukemia.

[0407] The tissue targeting compound of formula (I) of the present invention and / or a combination of a tissue targeting compound of formula (I) and another agent of the present invention preferably comprises a tissue targeting portion comprising a monoclonal antibody having binding affinity to prostate-specific membrane antigen (PSMA). PSMA expression in tumor cells has been observed in different types of diseases (particularly hyperproliferative diseases such as cancer) (Uijen et al., Eur. J. Nucl. Med. Mol. Imaging 2021, 48, 4350, doi: 10.1007 / s00259-021-05433-w). Examples of diseases and / or tumors suitable for treatment using a preferred tissue targeting compound of formula (I) of the present invention and / or a combination of a tissue targeting compound of formula (I) of the present invention include prostate cancer, salivary gland cancer, glioblastoma, brain cancer, thyroid cancer, hepatocellular carcinoma, and clear cell renal carcinoma.

[0408] In some embodiments, the present invention includes methods of treating a disease using a tissue targeting compound of formula (I) and / or a combination comprising a tissue targeting compound of formula (I) and another agent of the present invention, more preferably a hyperproliferative disease, more particularly, wherein the hyperproliferative disease is cancer, even more particularly, wherein the cancer disease is prostate cancer. In some embodiments, the present invention includes methods of treating a disease using a tissue targeting compound of formula (I) and / or a combination comprising a tissue targeting compound of formula (I) and another agent of the present invention, preferably a hyperproliferative disease, more preferably a cancer, characterized by overexpression of PSMA in tumor cells.

[0409] In some embodiments, the present invention includes methods of treating a hyperproliferative disease, more specifically cancer, more specifically prostate cancer, comprising administering to a subject in need thereof an effective amount of at least one tissue targeting compound of formula (I) and / or a combination comprising a tissue targeting compound of formula (I) and another agent of the present invention. In some embodiments, the present invention includes methods of treating a hyperproliferative disease characterized by overexpression of PSMA in tumor cells, more specifically, wherein the hyperproliferative disease is cancer, more specifically prostate cancer, comprising administering to a subject in need thereof an effective amount of at least one tissue targeting compound of formula (I) and / or a combination comprising a tissue targeting compound of formula (I) and another agent of the present invention.

[0410] In some embodiments, the present invention provides a tissue targeting compound of formula (I) as described above, and / or a combination comprising a tissue targeting compound of formula (I) as described above and another agent of the present invention, for use in treating and / or preventing a disease, in particular a hyperproliferative disease, more particularly cancer, even more particularly, wherein the cancer disease is prostate cancer. In some embodiments, the present invention includes a tissue targeting compound of formula (I) as described above, and / or a combination comprising a tissue targeting compound of formula (I) as described above and another agent of the present invention, for use in treating and / or preventing a hyperproliferative disease characterized by overexpression of PSMA in tumor cells.

[0411] In some embodiments, the present invention includes a tissue-targeted compound of formula (I) and / or a combination of a tissue-targeted compound of formula (I) and another agent of the present invention, for use in a method of inhibiting cell proliferation and / or inducing cell apoptosis, the method comprising contacting the cell with a tissue-targeted compound of formula (I) and / or a combination of a tissue-targeted compound of formula (I) and another agent. In some embodiments, the present invention includes a tissue-targeted compound of formula (I) and / or a combination of a tissue-targeted compound of formula (I) and another agent of the present invention, for use in a method of inhibiting tumor cell proliferation and / or inducing tumor cell apoptosis, the method comprising contacting tumor cells with a tissue-targeted compound of formula (I) and / or a combination of a tissue-targeted compound of formula (I) and another agent, wherein PSMA is overexpressed in tumor cells.

[0412] In some embodiments, the present invention includes a tissue-targeted compound of formula (I) and / or a combination comprising a tissue-targeted compound of formula (I) and another agent for use in a method of treating a hyperproliferative disease, more specifically, wherein the hyperproliferative disease is cancer, even more specifically, wherein the cancer disease is prostate cancer. In some embodiments, the present invention includes a tissue-targeted compound of formula (I) and / or a combination comprising a tissue-targeted compound of formula (I) and another agent for use in a method of treating a hyperproliferative disease characterized by overexpression of PSMA in tumor cells.

[0413] In some embodiments, the present invention includes the use of a tissue targeting compound of formula (I) and / or a combination comprising a tissue targeting compound of formula (I) and another agent in the preparation of a medicament for treating and / or preventing a hyperproliferative disease. In some embodiments, the present invention includes the use of a tissue targeting compound of formula (I) and / or a combination comprising a tissue targeting compound of formula (I) and another agent in the preparation of a medicament for treating and / or preventing a hyperproliferative disease characterized by overexpression of PSMA in tumor cells.

[0414] In some embodiments, the present invention includes the use of a tissue targeting compound of formula (I) and / or a combination comprising a tissue targeting compound of formula (I) and another agent in the preparation of a medicament for treating a hyperproliferative disease, particularly cancer, more particularly prostate cancer.

[0415] Drug administration

[0416] In a combination comprising a tissue targeting compound of formula (I) and another agent, the tissue targeting compound of formula (I) and the other agent may be administered sequentially in any order, or simultaneously.

[0417] The tissue targeting compound of formula (I) or the combination of the present invention comprising the tissue targeting compound of formula (I) and another agent can be administered in any form by any effective route, including, for example, oral, parenteral, enteral, intravenous, intraperitoneal, topical, transdermal (e.g., using any standard patch), ocular, nasal, topical, non-oral (e.g., aerosol), inhalation, subcutaneous, intramuscular, buccal, sublingual, rectal, vaginal, intra-arterial, and intrathecal, etc. They can be administered alone or in combination with any active or inactive ingredient. Preferably, the tissue targeting compound of formula (I) or the combination of the present invention is administered intravenously.

[0418] The tissue targeting compound of formula (I) or the combination of the present invention comprising the tissue targeting compound of formula (I) and another pharmaceutical agent can be converted into conventional pharmaceutical preparations in a known manner, which can be liquid or solid preparations, such as but not limited to plain and enteric-coated tablets, capsules, pills, powders, granules, elixirs, tinctures, solutions, suspensions, syrups, solid and liquid aerosols and emulsions.

[0419] The relative proportions of each compound in the combination of the invention comprising a tissue targeting compound of formula (I) and another agent may also be selected based on their respective mechanisms of action and disease biology. The relative proportions of each compound may vary widely.

[0420] Where appropriate, the release of one or more agents in the combination of the invention comprising a tissue targeting compound of formula (I) and another agent may also be controlled to provide the desired therapeutic activity in a single dosage form, combination pack, kit or in separate independent dosage forms.

[0421] The present invention includes a pharmaceutical composition consisting of a pharmaceutically acceptable carrier and a pharmaceutically effective amount of a tissue targeting compound of formula (I) or a combination of the present invention (i.e., a tissue targeting compound of formula (I) as described above and another agent). A pharmaceutically acceptable carrier is any carrier that is relatively nontoxic and harmless to the patient at a concentration consistent with the effective activity of the active ingredient, so that any side effects caused by the carrier do not diminish the beneficial effects of the active ingredient. A pharmaceutically effective amount of a compound is an amount that produces a result or exerts an effect on the specific condition being treated.

[0422] For oral administration, the tissue targeting compound of formula (I) or the combination of the present invention comprising the tissue targeting compound of formula (I) and another agent can be formulated into solid or liquid preparations, such as solid dispersions, capsules, pills, tablets, lozenges, lozenges, melts, powders, solutions, suspensions or emulsions, and can be prepared according to methods known in the art for preparing pharmaceutical compositions. The solid unit dosage form can be a capsule, which can be a common hard shell or soft shell gelatin type, containing, for example, surfactants, lubricants and inert fillers, such as lactose, sucrose, calcium phosphate and corn starch.

[0423] If administered in tablet form, the tissue targeting compound of formula (I) or the combination of the invention comprising the tissue targeting compound of formula (I) and another agent can be tableted together with conventional tablet bases (e.g., lactose, sucrose, and corn starch) and binders (e.g., gum arabic, corn starch, or gelatin), disintegrants (e.g., potato starch, alginic acid, corn starch, and guar gum, tragacanth gum, gum arabic) to help tablet breakage and dissolution after administration, lubricants (e.g., talc, stearic acid or magnesium stearate, calcium stearate, or zinc stearate) to improve tablet granule flowability and prevent tablet material from adhering to the surface of tablet molds and punches, dyes, colorants, and flavorings (e.g., mint, wintergreen oil, or cherry flavoring) to enhance the aesthetic quality of the tablets and make them more acceptable to patients. Excipients suitable for oral liquid dosage forms include dicalcium phosphate and diluents, such as water and alcohols, such as ethanol, benzyl alcohol, and polyvinyl alcohol, with or without the addition of pharmaceutically acceptable surfactants, suspending agents, or emulsifiers. Various other materials may also be present as coatings or to otherwise modify the physical form of the dosage unit. For example, tablets, pills, or capsules may be coated with shellac, sugar, or both.

[0424] Dispersible powders and granules are suitable for preparing aqueous suspensions. They provide the active ingredient mixed with a dispersant or wetting agent, a suspending agent and one or more preservatives. Suitable dispersants or wetting agents and suspending agents are illustrated by those examples already mentioned above. Additional excipients such as those sweeteners, flavorings and coloring agents described above may also be present.

[0425] The tissue targeting compound of formula (I) or the combination of the present invention comprising the tissue targeting compound of formula (I) and another agent may also be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil such as liquid paraffin or a mixture of vegetable oils. Suitable emulsifiers may be (1) gums of natural origin such as gum arabic and tragacanth, (2) phospholipids of natural origin such as soybean and lecithin, (3) esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, (4) condensation products of the partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion may also contain sweeteners and flavoring agents.

[0426] Oily suspensions can be prepared by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspension may contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. The suspension may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate; one or more coloring agents; one or more flavoring agents; and one or more sweeteners such as sucrose or saccharin.

[0427] Syrups and elixirs may be formulated with sweetening agents such as glycerol, propylene glycol, sorbitol or sucrose. Such preparations may also contain a demulcent and preservative (such as methyl and propylparabens) as well as flavoring and coloring agents.

[0428] The tissue targeting compound of formula (I) or a combination of the present invention comprising a tissue targeting compound of formula (I) and another agent can also be administered parenterally, i.e., as an injectable dose of the compound in a physiologically acceptable diluent and a pharmaceutical carrier which can be a sterile liquid or liquid mixture, such as water, saline, aqueous dextrose and related sugar solutions and alcohols such as ethanol, isopropanol or hexadecyl alcohol, glycerols such as propylene glycol or polyethylene glycol, glycerol ketals such as 2,2-dimethyl-1,1-dioxolane-4-methanol, ethers such as poly(ethylene glycol) 400, oils, fatty acids, fatty acid esters or fatty acid glycerides or acetylated fatty acid glycerides, with or without the addition of pharmaceutically acceptable surfactants such as soaps or detergents, suspending agents such as pectin, carbomer, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifiers and other pharmaceutically acceptable adjuvants.

[0429] Preferably, the tissue targeting compound of formula (I) or the combination of the present invention comprising the tissue targeting compound of formula (I) and another agent is administered intravenously and / or orally together or separately. Preferably, in the combination of the present invention comprising the tissue targeting compound of formula (I) and another agent, the tissue targeting compound of formula (I) is administered intravenously and the other agent is administered orally.

[0430] Examples of oils that can be used for parenteral preparations of the present invention are oils of petroleum, animal, plant or synthetic origin, such as peanut oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, vaseline and mineral oil. Suitable fatty acids include oleic acid, stearic acid, isostearic acid and myristic acid. Suitable fatty acid esters are, for example, ethyl oleate and isopropyl myristate. Suitable soaps include fatty acid alkali metal salts, ammonium salts and triethanolamine salts, and suitable detergents include cationic detergents, such as dimethyldialkylammonium halides, alkylpyridinium halides and alkylamine acetates; anionic detergents, such as alkyl, aryl and olefin sulfonates, alkyl, olefins, ethers and monoglycerides of sulfate, and sulfosuccinates; nonionic detergents, such as fatty amine oxides, fatty acid alkanolamides and poly (ethylene oxide-propylene oxide) or ethylene oxide or propylene oxide copolymers; and amphoteric detergents, such as alkyl-β-aminopropionates and 2-alkylimidazoline quaternary ammonium salts, and mixtures.

[0431] Examples of surfactants used in parenteral formulations are polyethylene sorbitan fatty acid esters, such as sorbitan monooleate and the high molecular weight adducts of ethylene oxide with a hydrophobic base formed by the condensation of propylene oxide with propylene glycol.

[0432] The tissue targeting compound of formula (I) or the combination of the present invention comprising the tissue targeting compound of formula (I) and another agent can be in the form of a sterile injectable aqueous suspension. Such suspensions can be prepared according to known methods using suitable dispersants or wetting agents and suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinyl pyrrolidone, tragacanth gum and gum arabic; dispersants or wetting agents, which can be naturally occurring phospholipids, such as lecithin, condensation products of alkylene oxides with fatty acids, such as polyethylene glycol stearate, condensation products of ethylene oxide with long-chain fatty alcohols, such as heptadeca-ethyleneoxycetanol, condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol, such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyoxyethylene sorbitol monooleate.

[0433] Aseptic injection preparation can also be a sterile injection solution or suspension in a nontoxic parenteral acceptable diluent or solvent. Operable diluents and solvents are, for example, water, Ringer's solution, isotonic sodium chloride solution and isotonic glucose solution. In addition, sterile fixed oils are generally used as solvents or suspension media. For this purpose, any gentle fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can also be used to prepare injections.

[0434] The tissue targeting compound of formula (I) or the combination of the present invention comprising the tissue targeting compound of formula (I) and another agent can also be administered in the form of a suppository for rectal administration. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature and therefore melts in the rectum and releases the drug. Such materials are, for example, cocoa butter and polyethylene glycol.

[0435] Controlled-release preparations for parenteral administration include liposome microspheres, polymer microspheres, and polymer gel preparations known in the art.

[0436] The tissue targeting compound of formula (I) or the combination of the present invention comprising the tissue targeting compound of formula (I) and another agent can also be in the form of a solid dispersion. The solid dispersion can be a solid solution, a glass solution, a glass suspension, an amorphous precipitate in a crystalline carrier, a co-crystal or a single crystal, a compound or a complex formation, and a combination thereof.

[0437] As needed or desired, the tissue targeting compound of formula (I) or the combination of the present invention comprising the tissue targeting compound of formula (I) and another agent may also contain other conventional pharmaceutically acceptable ingredients, usually referred to as carriers or diluents. Such compositions in appropriate dosage forms may be prepared using conventional methods.

[0438] Common pharmaceutical ingredients that may be suitably used to formulate the composition for its intended route of administration include:

[0439] Acidifying agents (examples include but are not limited to acetic acid, citric acid, fumaric acid, hydrochloric acid, nitric acid);

[0440] Alkalizing agents (examples include, but are not limited to, ammonia solution, ammonium carbonate, diethanolamine, monoethanolamine, potassium hydroxide, sodium borate, sodium carbonate, sodium hydroxide, triethanolamine, trolamine);

[0441] Adsorbents (examples include, but are not limited to, powdered cellulose and activated carbon);

[0442] Aerosol propellants (examples include, but are not limited to, carbon dioxide, CCl2F2, F2ClC-CClF2, and CClF3);

[0443] Air displacers (examples include, but are not limited to, nitrogen and argon);

[0444] Antifungal preservatives (examples include, but are not limited to, benzoic acid, butylparaben, ethylparaben, methylparaben, propylparaben, sodium benzoate);

[0445] Antimicrobial preservatives (examples include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetylpyridinium chloride, chlorobutanol, phenol, phenylethyl alcohol, phenylmercuric nitrate, and thimerosal);

[0446] Antioxidants (examples include, but are not limited to, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, hypophosphorous acid, monothioglycerol, propyl gallate, sodium ascorbate, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium metabisulfite);

[0447] Binding materials (examples include, but are not limited to, block polymers, natural and synthetic rubbers, polyacrylates, polyurethanes, silicones, polysiloxanes, and styrene-butadiene copolymers);

[0448] Buffering agents (examples include, but are not limited to, potassium metaphosphate, dipotassium hydrogen phosphate, sodium acetate, anhydrous sodium citrate, and sodium citrate dihydrate);

[0449] Carriers (examples include, but are not limited to, acacia syrup, aromatic syrups, aromatic elixirs, cherry syrup, cocoa syrup, orange peel syrup, syrup, corn oil, mineral oil, peanut oil, sesame oil, antibacterial sodium chloride injection, and antibacterial water for injection);

[0450] Chelating agents (examples include, but are not limited to, edetate disodium and edetic acid);

[0451] Colorants (examples include, but are not limited to, FD&C Red No. 3, FD&C Red No. 20, FD&C Yellow No. 6, FD&C Blue No. 2, D&C Green No. 5, D&C Orange No. 5, D&C Red No. 8, caramel, and red iron oxide);

[0452] Clarifying agents (examples include, but are not limited to, bentonite);

[0453] Emulsifiers (examples include, but are not limited to, gum arabic, cetomacrogol, cetyl alcohol, glyceryl monostearate, lecithin, sorbitan monooleate, polyoxyethylene 50 monostearate);

[0454] Encapsulating agents (examples include, but are not limited to, gelatin and cellulose acetate phthalate);

[0455] Flavoring agents (examples include, but are not limited to, anise oil, cinnamon oil, cocoa, menthol, orange oil, peppermint oil, and vanillin);

[0456] Humectants (examples include, but are not limited to, glycerin, propylene glycol, and sorbitol);

[0457] Abrasives (examples include, but are not limited to, mineral oil and glycerin);

[0458] Oils (examples include, but are not limited to, arachis oil, mineral oil, olive oil, peanut oil, sesame oil, and vegetable oil);

[0459] ointment bases (examples include, but are not limited to, lanolin, hydrophilic ointment, polyethylene glycol ointment, petrolatum, hydrophilic petrolatum, white ointment, yellow ointment, and rose water ointment);

[0460] Penetration enhancers (transdermal delivery) (examples include, but are not limited to, monohydric or polyhydric alcohols, mono- or polyvalent alcohols, saturated or unsaturated fatty alcohols, saturated or unsaturated fatty esters, saturated or unsaturated dicarboxylic acids, essential oils, phosphatidyl derivatives, cephalins, terpenes, amides, ethers, ketones and ureas);

[0461] Plasticizers (examples include, but are not limited to, diethyl phthalate and glycerol);

[0462] Scavenging compounds that reduce and / or prevent radiolysis;

[0463] Solvents (examples include, but are not limited to, ethanol, corn oil, cottonseed oil, glycerol, isopropyl alcohol, mineral oil, oleic acid, peanut oil, purified water, water for injection, sterile water for injection, and sterile water for irrigation);

[0464] • Hardening agents (examples include, but are not limited to, cetyl alcohol, cetyl ester wax, microcrystalline wax, paraffin, stearyl alcohol, white wax, and yellow wax);

[0465] Suppository bases (examples include, but are not limited to, cocoa butter and polyethylene glycol (mixture));

[0466] Surfactants (examples include, but are not limited to, benzalkonium chloride, nonoxynol 10, oxtoxynol 9, polysorbate 80, sodium lauryl sulfate, and sorbitan monopalmitate);

[0467] Suspending agents (examples include, but are not limited to, agar, bentonite, carbomer, sodium carboxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, kaolin, methylcellulose, tragacanth, and magnesium aluminum silicate);

[0468] Sweeteners (examples include, but are not limited to, aspartame, glucose, glycerol, mannitol, propylene glycol, sodium saccharin, sorbitol, and sucrose);

[0469] Tablet anti-adherents (examples include, but are not limited to, magnesium stearate and talc);

[0470] Tablet binders (examples include, but are not limited to, gum arabic, alginic acid, sodium carboxymethylcellulose, compressible sugars, ethylcellulose, gelatin, liquid glucose, methylcellulose, non-cross-linked polyvinyl pyrrolidone, and pregelatinized starch);

[0471] Tablet and capsule diluents (examples include, but are not limited to, dibasic calcium phosphate, kaolin, lactose, mannitol, microcrystalline cellulose, powdered cellulose, precipitated calcium carbonate, sodium carbonate, sodium phosphate, sorbitol, and starch);

[0472] Tablet coating agents (examples include, but are not limited to, liquid dextrose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose, ethylcellulose, cellulose acetate phthalate, and shellac);

[0473] Tablet direct compression excipients (examples include but are not limited to calcium hydrogen phosphate);

[0474] Tablet disintegrants (examples include, but are not limited to, alginic acid, carboxymethylcellulose calcium, microcrystalline cellulose, polacrilin potassium, cross-linked polyvinyl pyrrolidone, sodium alginate, sodium starch glycolate, and starch);

[0475] Tablet glidants (examples include, but are not limited to, colloidal silicon dioxide, corn starch, and talc);

[0476] Tablet lubricants (examples include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, stearic acid, and zinc stearate);

[0477] Tablet / capsule opacifiers (examples include, but are not limited to, titanium dioxide);

[0478] Tablet polishing agents (examples include, but are not limited to, carnauba wax and white wax);

[0479] Thickeners (examples include, but are not limited to, beeswax, cetyl alcohol, and paraffin);

[0480] Tonicity agents (examples include, but are not limited to, dextrose and sodium chloride);

[0481] Viscosity-increasing agents (examples include, but are not limited to, alginic acid, bentonite, carbomer, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone, sodium alginate, and tragacanth); and

[0482] • Wetting agents (examples include, but are not limited to, heptadecaethyleneoxycetanol, lecithin, sorbitan monooleate, polyoxyethylene sorbitan monooleate, and polyethylene glycol stearate).

[0483] One skilled in the art can utilize the above information to utilize the present invention to its fullest extent. BRIEF DESCRIPTION OF THE DRAWINGS

[0484] Figure 1The heavy chain nucleotide sequence (SEQ ID NO.1) and the heavy chain deduced amino acid sequence (SEQ ID NO.2) of Pelgifatamab (IgG1 heavy chain cDNA gene).

[0485] Figure 2 .The light chain nucleotide sequence (SEQ ID NO.3) and the light chain deduced amino acid sequence (SEQ ID NO.4) of Pelgifatamab (κ light chain cDNA gene).

[0486] Figure 3 .The CDR heavy chain sequence (SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7) and CDR light chain sequence (SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.10) of Pelgifatamab.

[0487] Figure 4 Electrospray mass spectrometry of Macropa-Pelgifatamab ACC (antibody-chelator conjugate) (CAR 0.8)

[0488] Figure 5 Electrospray mass spectrometry of DOTA-Pelgifatamab ACC (antibody-chelator conjugate) (CAR 5.7)

[0489] Figure 6 Electrospray mass spectrometry of DOTA-Pelgifatamab ACC (antibody-chelator conjugate) (CAR 12.7)

[0490] Figure 7 .Size exclusion chromatograms of targeted actinium conjugates (TACs) Ac225-DOTA-Pelgifatamab (CAR 12.7), Ac225-DOTA-Pelgifatamab (CAR 5.7), and Ac225-Macropa-Pelgifatamab (CAR 0.8).

[0491] Figure 8 Radiochemical purity was defined as the radiochemical purity of 225 The ratio of the amount of actinium-225 bound to Ac-Pelgifatamab to the amount of actinium-225-labeled DOTA-J591.

[0492] Fig. 9Flow cytometry was used to measure the binding of the PSMA-targeting antibody Pelgifatamab, macropa-Pelgifatamab (non-radiolabeled ACC), and non-radiolabeled isotype control ACC to PSMA-expressing LNCaP prostate cancer cells.

[0493] Fig.10 . 225 Internalization of Ac-Pelgifatamab in LNCaP prostate cancer cells.

[0494] Fig.11 .Immune reaction fraction (IRF) of Ac225-Macropa-Pelgifatamab (CAR 0.8), Ac225-DOTA-Pelgifatamab (CAR5.7) and Ac225-DOTA-Pelgifatamab (CAR 12.7).

[0495] Fig.12 .Cytotoxicity of Ac225-Macropa-Pelgifatamab (CAR 0.8), Ac225-DOTA-Pelgifatamab (CAR5.7) and Ac225-DOTA-Pelgifatamab (CAR 12.7) against the PSMA-expressing prostate cancer cell line C4-2.

[0496] Fig.13 .Cytotoxicity of Ac225-Macropa-Pelgifatamab (CAR0.8), Ac225-DOTA-Pelgifatamab (CAR5.7) and Ac225-DOTA-Pelgifatamab (CAR12.7) against the PSMA-expressing prostate cancer cell line MDA-PCa-2b.

[0497] Fig.14 .Cytotoxicity of Ac225-Macropa-Pelgifatamab in different prostate cancer cell lines.

[0498] Fig.15 Biodistribution of Ac-225-macropa-Pelgifatamab (CAR 0.8) and Ac-225-DOTA-Pelgifatamab (CAR 12.7) in different tissues in the prostate cancer xenograft model MDA-PCA-2b. Animals received a total antibody dose of 0.75 mg / kg, 250 kBq / kg as a single intravenous injection

[0499] Fig.16.(a) Antitumor activity of Ac-225-macropa-Pelgifatamab (CAR 0.8) and Ac-225-DOTA-Pelgifatamab (CAR 12.7) in the Balb / c male nude mouse prostate cancer xenograft model C4-2, after a single intravenous injection of 300 kBq / kg, 0.14 mg / kg total antibody (b) Body weight change (percent, %) (c) White blood cell count at the end of the experiment.

[0500] Fig.17 . 225 In vivo mode of action of Ac-Pelgifatamab in the C4-2 model. (A) Untreated C4-2 tumors and those treated with 100 kBq / kg 225 Examples of γH2AX expression in C4-2 tumors treated with Ac-Pelgifatamab for (B) one day or (C) four days. Scale bar represents 200 μm. D. Untreated C4-2 tumors (as in panel A) and treated with 100 kBq / kg 225 γH2AX expression in C4-2 tumors treated with Ac-Pelgifatamab (n=3) for 1 day (as shown in panel B) or 4 days (as shown in panel C) was quantified using HSA software.

[0501] Fig.18 .Use a carrier or 225 Growth curves of LNCaP tumors in male SCID mice (n=10 / group) treated with Ac-Pelgifatamab (70, 125, or 250 kBq / kg; single dose, intravenous injection; total antibody dose 0.75 mg / kg). Statistical analysis was performed using two-way ANOVA and Tukey's test. ***, P<0.001 compared to vehicle (days 0-40).

[0502] Fig.19 . Use isotype control (300 kBq / kg; single dose, intravenous injection; total antibody dose 0.14 mg / kg) or 225 Growth curves of KUCaP-1 tumors in male scid / scid mice (n = 9–10 / group) treated with Ac-Pelgifatamab (70, 150, or 300 kBq / kg; single dose, intravenous injection; total antibody dose 0.75 mg / kg). Statistical analysis was performed using two-way ANOVA and Tukey's test. ***, P < 0.001 compared with vehicle (days 0–21); ###, P < 0.001 compared with isotype control (days 0–21); §§, P < 0.001 compared with isotype control (days 0–21). 225 Ac-Pelgifatamab, 300 kBq / kg, P < 0.01; §§§, compared with225 Ac-Pelgifatamab, 300 kBq / kg vs. P<0.001 (days 0-21); $$$, vs. 225 Ac-Pelgifatamab, 150 kBq / kg vs. P<0.001 (Days 0-21).

[0503] Fig. 20 .Use a carrier or 225 Ac-Pelgifatamab (300 kBq / kg; single dose, intravenous injection; total antibody dose 0.75 mg / kg) was used to treat male scid / scid mice (n = 9-10 / group) with medium-sized and large KUCaP-1 tumors (mean volume 470 mm at the start of treatment) 3 )’s growth curve.

[0504] Fig.21 . (E) The volume of KUCaP-1 tumors on the last day (day 21) of the isotype control group. Statistical analysis was performed using two-way ANOVA and Tukey's test. ***, P < 0.001 compared with vehicle (days 0-21); ###, P < 0.001 compared with isotype control (days 0-21); §§, P < 0.001 compared with 225 Ac-Pelgifatamab, 300 kBq / kgP<0.01;§§§, compared with 225 Ac-Pelgifatamab, 300 kBq / kg vs. P<0.001 (days 0-21); $$$, vs. 225 Ac-Pelgifatamab, 150 kBq / kg vs. P<0.001 (Days 0-21).

[0505] Fig. 22 Growth curves of 22Rv1 tumors treated with vehicle, darolutamide (100 mg / kg, BID, oral) and / or 225Ac-Pelgifatamab75 (A, upper panel) or 150 kBq / kg (B, lower panel), single dose, intravenous injection, total protein dose 0.75 mg / kg.

[0506] Fig.23 Darolutamide enhances PSMA expression, and combining it with 225Ac-pelgifatamab improves efficacy in prostate cancer cells.

[0507] A.–B. Flow cytometry analysis of cell surface PSMA expression in (A) VCaP cells and (B) 22Rv1 cells treated with increasing concentrations of darolutamide for 7 days. DMSO served as a baseline control, indicated by the dotted line.

[0508] C.–H. RNA expression of (CD)FOLH1, (EF)CDKN1A, and (GH)XRCC2 in 22Rv1 cells was measured by RT-qPCR after treatment with 2 μM darolutamide, 0.15 kBq / mL 225Ac-Pelgifatamab, or the combination of the two for 24 or 48 hours. Statistical analysis was performed using one-way analysis of variance (ANOVA) with Tukey's multiple comparison test and correction. *, p < 0.05 compared with untreated cells; **, p < 0.01 compared with untreated cells.

[0509] I.–J. Isobolograms of the combined effects of darolutamide and 225Ac-Pelgifatamab on the proliferation of (E) VCaP and (F) 22Rv1 cells. CI, combination index

[0510] Fig.24 .225Ac-Pelgifatamab and darolutamide antitumor efficacy in the ST1273 prostate cancer PDX model.

[0511] A. PSMA expression in ST1273 tumor xenografts (untreated) detected by IHC using PSMA antibody GCP-04. Darker colors indicate PSMA staining. Scale bar is 50 μm.

[0512] B. Growth of ST1273 tumors in mice treated with vehicle, 225Ac-Pelgifatamab (single dose, 75 or 150 kBq / kg, iv), darolutamide (100 mg / kg, BID, po), or their combination.

[0513] C. The volume of a single ST1273 tumor on day 30 is shown in (B). Statistical analysis was performed using one-way ANOVA and Tukey's test. ***, p<0.001 compared to vehicle. #, p<0.05 compared to darolutamide monotherapy.

[0514] D. Relative body weight changes of mice during the study are shown in (B).

[0515] Fig.25 .Antitumor efficacy of 225Ac-Pelgifatamab and darolutamide in the 22Rv1 prostate cancer model.

[0516] A. PSMA expression in 22Rv1 tumor xenografts (untreated) detected by IHC using PSMA antibody GCP-04. Darker colors indicate PSMA staining. Scale bar is 50 μm.

[0517] B. Growth of 22Rv1 tumors in mice treated with vehicle, isotype control (300 mg / kg, single dose, iv), 225Ac-Pelgifatamab (75, 150 or 300 kBq / kg, single dose, iv), darolutamide (100 mg / kg, BID, po), or their combination.

[0518] C. The volume of a single 22Rv1 tumor at day 16 is shown in (B). Statistical analysis was performed using one-way ANOVA and Tukey's test. **, p < 0.01 compared to vehicle; ***, p < 0.001 compared to vehicle. ###, p < 0.001 compared to darolutamide monotherapy. §, p < 0.05 compared to the corresponding 225Ac-Pelgifatamab monotherapy.

[0519] D. Relative body weight changes of mice during the study are shown in (B).

[0520] E. PSMA expression was determined by flow cytometry in isolated 22Rv1 cancer cells obtained from 22Rv1 tumor-bearing mice (n=2) treated with vehicle or darolutamide 18 days before sacrifice.

[0521] Fig.26 .Ac-225-Macropa-J592 and Ac-225-DOTA-J592 radiolabeled free chelator iTLC showing the percentage of radiolabeled free chelator (RFC) at t=0 h and after 96 h.

[0522] Experimental Section

[0523] Chemical names were generated using BIOVIADraw 2019 from Dassault Systems. In some cases, recognized names of commercially available reagents were used instead of the BIOVIADraw generated names.

[0524] Table 1 below lists the abbreviations used in this paragraph and the Examples section which are not explained in the text. Other abbreviations themselves have the meanings customary to the skilled person.

[0525] Table 1: Abbreviations

[0526] The following table lists the abbreviations used in this article.

[0527] 225 Ac Actinium-225

[0528] Ac-225 Actinium-225

[0529] ACC Antibody-Chelator Conjugate

[0530] ACN Acetonitrile

[0531] BCA Bicinchoninic acid

[0532] BRFF-HPC1 serum-free medium

[0533] CAR chelator to antibody ratio

[0534] DCTA 1,2-Diaminocyclohexanetetraacetic acid

[0535] DMA N,N-Dimethylacetamide

[0536] DMSO Dimethyl sulfoxide

[0537] DOTA 1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid

[0538] ESI Electrospray ionization

[0539] EtOH

[0540] FBS Fetal Bovine Serum

[0541] FPLC fast protein liquid chromatography

[0542] HCl

[0543] HPGe High Purity Germanium

[0544] HPLC High Performance Liquid Chromatography

[0545] iTLC instant thin layer chromatography

[0546] IRF immune response score

[0547] mAb monoclonal antibody

[0548] MEM-NEAA Minimum Essential Moderate Non-Essential Amino Acids

[0549] min

[0550] MS

[0551] NaCl Sodium chloride

[0552] nm nanometer

[0553] nmol nanomole

[0554] pelgi Pelgifatamab, prostate-specific membrane antigen IgG1 antibody

[0555] Pelgifatamab prostate-specific membrane antigen IgG1 antibody

[0556] PBS Phosphate Buffered Saline

[0557] PSMA Prostate-specific membrane antigen

[0558] RAC radioactivity concentration

[0559] RCP radiochemical purity

[0560] RPMI Roswell Park Memorial Institute 1640 Medium

[0561] SEC Size Exclusion Chromatography

[0562] TAC Targeted Actinium Conjugate (Ac-225 Labeled ACC)

[0563] TFA Trifluoroacetic acid

[0564] TOF Time of Flight

[0565] UPLC Ultra-Performance Liquid Chromatography

[0566] UV

[0567] All reagents whose synthesis is not described in the experimental section are either commercially available or are known compounds or can be formed from known compounds by methods known to those skilled in the art.

[0568] Various aspects of the invention described in this application are illustrated by the following examples, which are not meant to limit the invention in any way.

[0569] The exemplary test experiments described herein serve to illustrate the invention, and the invention is not limited to the examples given.

[0570] Tissue-targeted compounds containing Pelgifatamab

[0571] Characterization of ACC by size exclusion chromatography and mass spectrometry

[0572] The CAR of ACC was determined by SEC-MS (Water Acquity HPLC connected to Waters XEVO TOF; running buffer: 50 / 50 / 0.1 (v / v / v) ACN / water / TFA; flow rate: 0.06 mL / min, online desalting column: Waters Acquity BEH SEC, 1.7 μm, 2.1 x 150 mm; ESI+ mode) and expressed as the percentage of the MS peak height to the main peak height of the components mAb, mAb+1 chelator, mAb+2 chelator, mAb+3 chelator, etc., and using the formula CAR=Sum(n*An) / Sum An, where n equals the number of chelators and An equals the strength of the antibody conjugate bound to n chelators. The purity and concentration of ACC were determined by SEC-UV method (Agilent 1260 Infinity HPLC system, running buffer: 10% DMSO / PBS; flow rate: 0.3 mL / min, column: Waters Acquity BEH SEC, 1.7 μm, 4.6 x 300 mm, detection: UV at 280 nm). This method separates higher polymers, dimers and fragments from ACC monomers.

[0573] Example 1: Preparation of macropa-Pelgifatamab ACC

[0574] 6-[[16-[(6-carboxy-2-pyridinyl)methyl]-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane-7-yl]methyl]-4-[2-(4-isothiocyanatophenyl)ethoxy]pyridine-2-carboxylic acid (5.8 mg, 8,2 μmol) was dissolved in DMA (728 μL) and then added to Pelgifatamab (493 mg, 3.3 μmol) in PBS (25 mL). The solution was adjusted to pH 9 by adding 1 M carbonate buffer (pH 9.5, (5 mL)) and the solution was shaken overnight. The product was purified by FPLC (column: HiLoad 16 / 600 Superdex 200pg column; running buffer: 100mM acetate / 100mM NaCl 1:1, pH 5; flow rate: 1mL / min; detection: UV 214 / 254nm) to obtain 386mg (77% yield) of macropa-Pelgifatamab ACC in 100mM acetate / 100mM NaCl 1:1 (2.3mg / mL). The CAR was determined to be 0.8 by MS. The monomer purity was determined to be 99% by SEC-UV.

[0575] Example 2: Preparation of DOTA-Pelgifatamab ACC CAR 5.7

[0576] p-SCN-Bn-DOTA (macrocyclic compound, B-205) was dissolved in PBS at a concentration of 10 mg / mL. 261 μL was added to 990 μL Pelgifatamab (10 mg / mL) dissolved in PBS. The pH was adjusted to 9 with 1M carbonate buffer. The mixture was incubated on a thermomixer at 37°C with shaking at 550 rpm for 2 hours. The conjugate was purified by FPLC (column: HiLoad 16 / 600Superdex 200pg column; running buffer: 100mM acetate / 100mM NaCl 1:1, pH 5; flow rate: 1mL / min; detection: UV 280nm). The concentration of the combined DOTA-Pelgifatamab fractions was measured by SEC-UV to be 2.6 mg / mL. The CAR was determined to be 5.7 by MS. The monomer purity was determined to be 99% by SEC-UV.

[0577] Example 3: Preparation of DOTA-Pelgifatamab ACC CAR 12.7

[0578] p-SCN-Bn-DOTA (macrocyclic compound, B-205) was dissolved in PBS at a concentration of 10 mg / mL. 470 μL was added to 990 μL Pelgifatamab (10 mg / mL) dissolved in PBS. The pH was adjusted to 9 with 1M carbonate buffer. The mixture was shaken at 550 rpm on a thermomixer at 22°C to incubate overnight in the dark. The conjugate was purified by FPLC (column: HiLoad 16 / 600Superdex200pg column; running buffer: 100mM acetate / 100mM NaCl 1:1, pH 5; flow rate: 1mL / min; detection: UV 280nm). The concentration of the combined DOTA-Pelgifatamab fractions was measured to be 2.5 mg / mL by BCA protein assay. CAR was determined to be 12.7 by MS. The monomer purity was 98% by SEC-UV.

[0579] Radiolabeling

[0580] Macropa-Pelgifatamab, DOTA-Pelgifatamab CAR=5.7 and DOTA-Pelgifatamab CAR=12.7 were radiolabeled with Ac-225 with a specific activity of 2MBq / mg and diluted with 0.1M acetate buffer pH 5 to a radioactivity concentration of 1.5kBq / μL. For macropa-Pelgifatamab, the mixture was incubated at room temperature for one hour, and two DOTA-Pelgifatamab ACCs were incubated on a heating block at 60°C for one hour. The RCP of TAC was measured by instant thin layer chromatography, eluted with citrate buffer, and each sample was repeated three times. Unlabeled Ac-225 eluted at the solvent front, while the radiolabeled compound did not move from the application part. Radioactivity measurements were completed with a digital gamma-ray spectrometer (DSPEC-50, Ortec). Activity was measured at least 6 hours after radiolabeling to allow the daughter nuclide to reach equilibrium with Ac-225. The RCPs of the samples (defined as [(radioactivity in the applied portion) / (total radioactivity on the test strip)] x 100) were 99.9% ± 0.013; 99.2% ± 0.049 and 99.5% ± 0.110 for Ac-225-macropa-Pelgifatamab, Ac-225-DOTA-Pelgifatamab CAR 5.7 and Ac-225-DOTA-Pelgifatamab CAR 12.7, respectively.

[0581] Pelgifatamab-macropa and PSMA-binding antibody J591 conjugated with DOTA were both labeled with actinium-225. Labeling was performed at room temperature. J591-DOTA was also labeled at 60°C. Radiochemical purity was determined by instant thin layer chromatography (iTLC). Labeled and free actinium-225 were counted in a germanium detector using the actinium-225 specific energy line. Figure 8 ) was more efficient at room temperature. After radiolabeling, the radiochemical purity of 225Ac-Pelgifatamab was determined to be 100% ( Figure 8 ).

[0582] Size Exclusion Chromatography of TAC

[0583] The purity of TAC was determined by SEC-UV. This method separates higher aggregates, dimers and fragments from ACC monomers. The separation was performed using Acquity UPLC Protein BEH SEC, 1.7 μm, 4.6 x 300 mm column at 30°C. The mobile phase was 170 mM ammonium acetate and 300 mM NaCl containing 10% 2-propanol and 0.1 mM DCTA. The flow rate was 0.3 mL / min and ACC was detected at UV 280 nm. The analysis was performed on an Agilent 1200 series HPLC with an injection volume of 35 μL.

[0584] Table 2 Purity of TAC determined by SEC-UV

[0585]

[0586] Immune Reaction Fraction (IRF)

[0587] Soon after radiolabeling was completed, the binding of each sample to PSMA-coated M-270-carboxyl Dynabeads was determined. Briefly, 250 μg of beads were placed in 50 μL of buffer (PBS / 3% BSA) in a 2 mL Eppendorf tube. The samples were repeated three times and included samples blocked with an excess of naked PSMA mAb. The blocking step lasted 40 minutes while shaking the tubes at 750 rpm on a thermomixer (Eppendorf) at room temperature. The radiolabeled compound was diluted to 5 Bq / μL in buffer and added to each tube containing beads. The binding lasted 80 minutes while shaking at 750 rpm at room temperature. After the binding step was completed, 40 μL of buffer was added to each tube and the beads were separated from half of the supernatant using a magnet (DynaMag-2). The radioactivity in the supernatant and beads of each sample was measured using a digital gamma-ray spectrometer. Activity was measured at least 6 hours later to allow the daughter nuclide to reach equilibrium with Ac-225. Total binding was defined as [(activity in beads - activity in supernatant) / (total activity)] x 100. Nonspecific binding was obtained by the same calculation as for the blocked samples, and IRF was defined as [specific binding - nonspecific binding]. For Ac-225-macropa-Pelgifatamab, Ac-225-DOTA-Pelgifatamab CAR 5.7, and Ac-225-DOTA-Pelgifatamab CAR 12.7, the IRFs were 95%, 92%, and 74%, respectively ( Fig.11 ).

[0588] Flow cytometry

[0589] The in vitro binding properties of the fully human PSMA-targeting antibody Pelgifatamab, the antibody-chelator conjugate (ACC) pelgi-macropa, and an isotype control were evaluated in PSMA-expressing LNCaP prostate cancer cells. EC50 The binding of PSMA-targeted antibody Pelgifatamab, macropa-Pelgifatamab (non-radiolabeled ACC), and non-radiolabeled isotype control ACC to PSMA-expressing LNCaP prostate cancer cells was determined by flow cytometry ( Fig. 9 ).

[0590] Internalization

[0591] After 2 hours of incubation, LNCaP cells were used to determine 225 Internalization of Ac-Pelgifatamab. The cell binding and internalization activity of actinium-225 were measured using a gamma counter (Wizard 2470, PerkinElmer). Fig.10 ).

[0592] Cytotoxicity

[0593] The in vitro cytotoxicity of Ac-225-macropa-Pelgifatamab, Ac-225-DOTA-Pelgifatamab CAR=5.7 and Ac-225-DOTA-Pelgifatamab CAR=12.7 was measured in the PSMA-expressing prostate cancer cell lines C4-2 and MDA-PCa-2b. Cells were seeded in cell culture medium (RPMI / 10% FBS / 1% HEPES / 1% MEM NEAA / 1% NaPyruvate / 1% L-glutamine and BRFF-HPC1 / 20% FBS, respectively) at appropriate cell densities. One day after seeding, Ac-225-labeled compounds with a specific activity of 2MBq / mg were titrated simultaneously with the relevant negative isotype controls radiolabeled under the same conditions. Cells were exposed to radioligands for two hours. Six days later, cell viability was determined using CellTiterGlo (Promega) and luminescence was measured on an EnVision plate reader. In C4-2 cells, the IC50 values ​​of Ac-225-macropa-Pelgifatamab, Ac-225-DOTA-Pelgifatamab CAR 5.7, and Ac-225-DOTA-Pelgifatamab CAR 12.7 were 0.09, 0.15, and 0.29 kBq / ml, respectively. Fig. 9), in MDA-PCa-2b cells, the IC50 values ​​of Ac-225-macropa-Pelgifatamab, Ac-225-DOTA-Pelgifatamab CAR 5.7 and Ac-225-DOTA-Pelgifatamab CAR 12.7 were 0.05, 0.09 and 0.2 kBq / ml, respectively ( Fig.10 The IC50 value of Ac-225-macropa-isotype was 256 kBq / ml in C4-2 cells and 3 kBq / ml in MDA-PCa-2b cells ( Fig.12 +13). The IC50 value of Ac-225-macropa-isotype was ∼256 kBq / ml in C4-2 cells and 3 kBq / ml in MDA-PCa-2b cells ( Fig.12 +13).

[0594] Cytotoxicity

[0595] The in vitro cytotoxicity of Ac-225-macropa-Pelgifatamab was measured in prostate cancer cell lines expressing different levels of PSMA. PC-3, DU-145, 22Rv1, C4-2 and LNCaP cells were seeded at appropriate cell densities in the following cell culture media: DMEM / Ham's F12+10% FCS; DMEM+10% FCS; RPMI+10% FCS; DMEM / Ham's F12+10% FCS+insulin (human) (5 μg / ml, Sigma#I9278), transferrin (human); (final concentration: 5 μg / ml), D-biotin (final concentration: 0.25 μg / ml, Sigma: B4639), adenine (final concentration: 25 μg / ml, Sigma: A9795), L-3,3,5-triiodothyronine sodium salt, Sigma: T6397 (final concentration: 13.6 pg / ml); RPMI+20% FCS, as recommended by each supplier. One day after inoculation, Ac-225-macropa-Pelgifatamab with a specific activity of 2 MBq / mg was titrated in parallel with the relevant negative isotope control radiolabeled under the same conditions. Cells were treated for 5 days and then cell viability was determined using CellTiterGlo (Promega). Strong cytotoxicity was observed in LNCaP, C4-2, and 22R1 cells, while almost no cytotoxicity was observed in PMSA-negative PC-3 and DU-145 cells ( Fig.14 ).

[0596] Biodistribution study of Ac-225-macropa-Pelgifatamab

[0597] The biodistribution of Ac-225-macropa-pelgifatamab was studied in the PSMA-expressing prostate cancer xenograft model MDA PCa 2b. Female RJ:NMRI-Foxn1nu / nu mice were implanted subcutaneously with testosterone pellets and inoculated on the right flank with 1×10 6 MDAPCa 2b cells. Animals were pretreated with 200 μg / mouse nonspecific IgG2a antibody 24 hours prior to heat administration. Animals were treated with 300 kBq / kg Ac-225-macropa-Pelgifatamab IV for a total protein dose of 0.75 mg / kg. In a separate in vivo study, the xenograft model MDA PCa 2b was treated with 250 kBq / kg Ac-225-DOTA-Pelgifatamab for a protein dose of 0.14 mg / kg. In both studies, animals were sacrificed at 72, 168, 336, and 504 hours after treatment, and 225Ac was measured in blood, tumors, liver, kidneys, spleen, and femurs. For Ac-225-macropa-Pelgifatamab-treated animals, the data indicated robust and sustained accumulation in tumors, whereas accumulation in normal organs was generally low and transient and cleared over time. Compared with animals treated with 225Ac-DOTA-Pelgifatamab, bone clearance was improved, radioactivity in the liver was low and decreased over time, and uptake in the spleen was reduced ( Fig.15 ). These data may explain the improved in vivo complex stability of 225Ac to macropa compared to DOTA.

[0598] Comparison of in vivo efficacy between Ac-225-macropa-Pelgifatamab and Ac-225-DOTA-Pelgifatamab

[0599] The compounds Ac-225-macropa-, Ac-225-DOTA-Pelgifatamab and isotype-macropa were all labeled with 225Ac. Mice transplanted with C4-2 prostate cancer cells were treated with a single IV dose of 300 kBq / kg, with a total antibody dose of 0.14 mg / kg. Fig.16 As shown in (A) and (B), Ac-225-macropa-Pelgifatamab had higher antitumor activity than Ac-225-DOTA-Pelgifatamab. The weight loss in all targeted therapy groups was less than 10%. The effect on the hematopoietic system was determined by white blood cell counts at the end of the experiment ( Fig.16(C)).

[0600] Immunohistochemistry

[0601] In use 120kBq / kg 225 C4-2 tumors were harvested 24 or 96 hours after Ac-Pelgifatamab (total antibody dose of 0.75 mg / kg) treatment and the levels of phosphorylated histone H2AX (γ-H2AX) were assessed in paraffin-embedded C4-2 tumor tissue sections (n=3). (17A) Untreated C4-2 tumors neutralized 100 kBq / kg Ac-Pelgifatamab (total antibody dose of 0.75 mg / kg) 225 Examples of γH2AX expression in C4-2 tumors treated with Ac-Pelgifatamab for one day (17B) or four days (17C). Scale bar represents 200 μm. 17D. Untreated C4-2 tumors (as in panel A) and treated with 100 kBq / kg 225 γH2AX expression in C4-2 tumors treated with Ac-Pelgifatamab (n=3) for 1 day (as shown in panel B) or 4 days (as shown in panel C) was quantified using HSA software ( Fig.17 A, B, C, D).

[0602] 225 In vivo antitumor efficacy of Ac-Pelgifatamab in LNCaP and KUCaP-1 prostate cancer models.

[0603] The antitumor efficacy of 225Ac-Pelgifatamab was evaluated in the LNCaP prostate cancer model: male SCID mice (6 weeks, 22 g, Janvier Labs) were implanted subcutaneously with testosterone pellets (12.5 mg, 4 mm) and then inoculated subcutaneously with 5x10 6 LNCaP cells. When the tumor reaches an average size of 160 mm 3 The mice were randomly divided into groups (10 mice per group) and two days later, they were injected intravenously with vehicle or a single dose of 225 Ac-Pelgifatamab (70, 125 or 250 kBq / kg, total antibody dose 0.75 mg / kg) was used for treatment. The study was terminated on day 112 after tumor cell inoculation. In the LNCaP model, 70, 125 and 250 kBq / kg (single dose) 225 Ac-Pelgifatamab showed very strong anti-tumor efficacy, with T / C 体积 The (treatment / control) ratio is 0.00 ( Fig.18 ).

[0604] In the KUCaP-1 efficacy study, male scid / scid mice (8 weeks, 22 g, Janvier Labs) were implanted subcutaneously with 5x5x5 mm KUCaP-1 tumor fragments. When the average tumor size reached 220 mm 3 At 4 pm, mice were randomly divided into groups (9-10 mice per group) and injected intravenously the next day with vehicle or a single dose of isotype control (300 kBq / kg, total antibody dose 0.14 mg / kg) or 70, 150 or 300 kBq / kg (total antibody dose 0.75 mg / kg) of 225 Ac-Pelgifatamab treatment. In addition, for one treatment group, tumors were allowed to reach 470 mm 3 size, and then a single dose of 300 kBq / kg 225 Ac-pelgi treatment. The study was terminated on day 72 after tumor cell inoculation. In the patient-derived KuCaP model, 75, 150, and 300 kBq / kg (single dose) 225 Ac-Pelgifatamab showed dose-dependent efficacy, resulting in T / C 体积 The ratios were 0.55, 0.44 and 0.26 respectively ( Fig.19 +21). It is noteworthy that even in mice with large tumors, a single dose of 300 kBq / kg 225 Ac-Pelgifatamab can also induce tumor regression ( Fig. 20 ).

[0605] Tissue-targeted compounds containing TLX592 / J592

[0606] Characterization of ACC by size exclusion chromatography and mass spectrometry

[0607] The CAR of ACC was determined by LCMS. For Ac225-Macropa-J592, the CAR was 0.75, and for Ac225-DOTA-J592, the CAR was 6.7.

[0608] Macropa-NCS-J592

[0609] 6-[[16-[(6-carboxyl-2-pyridinyl)methyl]-1,4,10,13-tetraoxa-7,16-diazacyclooctadecane-7-yl]methyl]-4-[2-(4-isothiocyanatophenyl)ethoxy]pyridine-2-carboxylic acid was dissolved in DMA to 10 mg / ml. Anti-J592 antibody was dissolved in PBS to 10 mg / mL and the pH was adjusted to 9 with 1 M carbonate buffer. The chelating agent and antibody were mixed and incubated on a thermomixer at room temperature with shaking at 350 rpm for 1 hour. The product was analyzed by FPLC (column: 200Increase10 / 300GL; running buffer: 6mM citrate / 10mM histidine / 10mM glycine / 100mM NaCl; flow rate: 1mL / min; detection: UV 214 / 254nm). Concentration and monomer purity were determined by SEC-UV, and CAR was determined by SEC-MS using positive electrospray ionization, with the following formula: CAR = [(A*An) / A]0, A = all signal intensities of the maximum entropy model (entropy envelope), An = signal of the maximum entropy model containing chelating agent. CAR was determined to be 0.8.

[0610] DOTA-NCS-J592

[0611] p-SCN-Bn-DOTA (macrocycle, B-205) and anti-J592 antibody were dissolved in PBS to 10 mg / mL. The pH of the antibody was adjusted to 9 with 1 M carbonate buffer. p-SCN-Bn-DOTA was added to the antibody and the mixture was incubated on a thermomixer and shaken at 350 rpm for 1 hour at room temperature. The conjugate was analyzed by FPLC (column: 200Increase 10 / 300GL; running buffer: 6mM citrate / 10mM histidine / 10mM glycine / 100mM NaCl; flow rate: 1mL / min; detection: UV 280nm) purification. The concentration and monomer purity of the combined DOTA-antibody fractions were measured by SEC-UV. CAR was determined by SEC-MS using positive electrospray ionization, and the formula is as follows: CAR = [(A*An) / A]0, A = all signal intensities of the maximum entropy model, An = the signal of the maximum entropy model containing chelating agent. CAR was determined to be 6.7.

[0612] Buffer exchange

[0613] Amicon Ultra-4 30K (Millipore, product number UFC803096, batch number R4JA84392) was used for buffer exchange of J592-DOTA and J592-Macropa. The ultrafiltration membrane in the Ultra-4 device was rinsed with 3.5 mL of 0.1 M acetate buffer (No. J145-046, 21Nov22AHJ). Spin at 7000×g for 2 minutes. Remove the acetate buffer. Add the sample and 3.5 mL of 0.1 M acetate buffer and spin at 7000×g for 10 minutes for buffer exchange. Recover the sample and dilute with 0.5 mL of 0.1 M acetate buffer. Determine the new concentration by comparing the UV signal of the previous Amicon pre-sample with the known concentration using SE-HPLC.

[0614] Radiolabeling

[0615] Macropa-J592 CAR 0.75 and DOTA-J592 CAR 6.7 were radiolabeled with Ac-225 with a specific activity of 2 MBq / mg and diluted to a radioactivity concentration of 1.5 kBq / μL with 0.1 M acetate buffer pH 5. The mixture was incubated at room temperature for one hour for J592-Macropa, while DOTA-J592 was incubated at 60°C on a heating block for one hour. The RCP of TAC was measured by instant thin layer chromatography, using citrate buffer for elution, repeated three times for DOTA and once for Macropa. Unlabeled Ac-225 was eluted at the solvent front, while the radiolabeled compound did not move from the applied part. Radioactivity measurements were performed with a HPGe detector (DSPEC-50, Ortec). The activity was measured at least 6 hours after radiolabeling to allow the daughter nuclide to reach equilibrium with Ac-225. Two time points were analyzed, namely 0 hours and after 96 hours of storage at room temperature. The RCP of the samples (defined as [(radioactivity in the applied portion) / (total radioactivity on the test paper)] x 100) were 99.9% (0 hour) and 99.8% (96 hours), respectively; for Ac-225-macropa-J592 and Ac225-DOTA-J592, they were 99.5% ± 0.06 (0 hour) and 96.0% ± 0.35 (96 hours), respectively. The data show that at t = 96 hours, Ac-225-macropa-J592 releases less 225Ac than Ac225-DOTA-J592 and is therefore more stable to radionuclide leakage.

[0616] Size Exclusion Chromatography of TAC

[0617] The purity of TAC was determined by SEC. This method separates higher aggregates, dimers, and fragments from ACC monomers. Separation was performed using Acquity UPLC Protein BEH SEC, 1.7 μm, 4.6x 300mm column at 30°C. The mobile phase was 200mM ammonium acetate and 300mM NaCl containing 10% 2-propanol and 0.1mM DCTA. The flow rate was 0.3mL / min and ACC was detected at UV 280nm. The analysis was performed on an Agilent 1200 series HPLC with an injection volume of 40μL.

[0618] Table 3. Purity of TAC detected by SEC-UV

[0619]

[0620] The results in Table 3 show that the tissue targeting compounds of formula (I) have improved stability compared to their DOTA counterparts as evidenced by the lower amounts of byproducts (ie, aggregates and dimers) present after 96 hours.

[0621] iTLC(RFC)

[0622] Free chelator iTLC was performed to measure the amount of free chelator in the conjugate solution over time. In brief, the iTLC.SG. plate was cut to obtain a strip of 12 cm x 1 cm. The application point, cut point, and run end point were marked as 1 cm, 4 cm, and 10 cm, respectively. 2 μL of TAC sample was applied to n=3 iTLC strips and eluted with a freshly prepared 1:1 1M ammonium acetate / 100% methanol mobile phase. These strips were developed in 3 mL of mobile phase in LSC vials. When the liquid front reached the marked end point, they were removed from the vial and placed on aluminum foil to dry. The sample was read after 6 hours to obtain a long-term equilibrium in the sample. The iTLC strips were measured using an iTLC scanner AR-2000 from Eckert & Siegler, with 1 second per strip at a voltage of 1000 volts.

[0623] Fig.26 The tissue targeting compounds of formula (I) were shown to have increased stability to decomplexation compared to their DOTA counterparts, as evidenced by the lower amount of free chelate present after 96 hours.

[0624] Combination study of 225Ac-Pelgifatamab and darolutamide

[0625] The effects of darolutamide and 225Ac-Pelgifatamab on in vitro PSMA mRNA and protein expression were studied in VCaP and 22Rv1 prostate cancer cells. To evaluate PSMA protein expression, VCaP and 22Rv1 cells were treated with 0.08-5 μM darolutamide for 7 days, and PSMA expression was determined by flow cytometry using mouse PSMA-specific monoclonal antibody (mAb) J591 (made in-house). To evaluate FOLH1 gene expression, 22Rv1 cells were treated with 2 μM darolutamide and / or 0.15 kBq / mL 225Ac-Pelgifatamab for 24 or 48 hours. RNA was extracted using the RNeasy Plus Mini Kit (Qiagen), transcribed into cDNA using SuperScript IV VILO Master Mix (Invitrogen), and analyzed by qPCR.

[0626] To evaluate the in vitro combined effects of 225Ac-Pelgifatamab and darolutamide, VCaP and 22Rv1 cells were treated with darolutamide, 225Ac-Pelgifatamab, or their combination for 5 days. Cell viability was measured by ELISA (Promega). Combination index (CI) was calculated according to the median effect model of Chou-Talalay (8), and synergy was defined as CI < 0.9.

[0627] In vitro, darolutamide induced a more than 10-fold and 2-fold increase in PSMA expression in androgen-dependent VCaP cells and androgen-independent 22Rv1 cells, respectively. Fig.23 ).

[0628] After 48 hours of treatment with darolutamide combined with 225Ac-Pelgifatamab, FOLH1 gene expression in 22Rv1 cells increased by 76% ( Fig.23 ). No increase was observed with either monotherapy.

[0629] 225Ac-Pelgifatamab monotherapy and 225Ac-Pelgifatamab combined with darolutamide increased the RNA expression of CDKN1A in 22Rv1 cells ( Fig.23 ) and decreased the expression of XRCC2 ( Fig.23 ).

[0630] Combination therapy of darolutamide and 225Ac-Pelgifatamab was synergistic in VCaP and 22Rv1 cells, with combination indices of 0.34 and 0.38, respectively. Fig.23 ).

[0631] Darolutamide enhances PSMA expression, and combining it with 225Ac-pelgifatamab improves efficacy in prostate cancer cells.

[0632] PSMA expression in isolated, untreated 22Rv1 and ST1273 tumors was determined by immunohistochemistry (IHC) in formalin-fixed, paraffin-embedded (FFPE) tissue sections using the murine PSMA-specific monoclonal antibody GCP-04 (Novus Biologicals).

[0633] The in vivo efficacy of 225Ac-Pelgifatamab in combination with darolutamide was investigated in the androgen-dependent ST1273 and androgen-independent 22Rv1 prostate cancer models in mice.

[0634] In the ST1273 study, female NMRI nude mice (n=10 / group) were implanted with ST1273 patient-derived tumor fragments and, 19 days later, treated with vehicle (PEG400, propylene glycol, glucose; v / v% 50:30:20), 225Ac-Pelgifatamab (75 or 150 kBq / kg, single dose, intravenous injection), darolutamide (100 mg / kg, twice daily (BID) for 34 days, oral administration), or their combination.

[0635] In the 22Rv1 study, male NMRI nude mice (n=10 / group) were inoculated with 22Rv1 cells and treated 15 days later with vehicle (acetate buffer), 225Ac-Pelgifatamab (75 kBq / kg, single dose, intravenous injection; total antibody dose 75 mg / kg), darolutamide (100 mg / kg, BID, oral), or their combination.

[0636] In vivo expression of PSMA protein was evaluated in 22Rv1 prostate tumor-bearing male nude mice (nu / nu) treated with vehicle or darolutamide (100 mg / kg, BID, oral) for 18 days. 22Rv1 tumors (n=2) were extracted and isolated tumor cells were analyzed by flow cytometry using J591 antibody.

[0637] Combination of 225Ac-Pelgifatamab and darolutamide enhances therapeutic efficacy in the androgen-dependent ST1273 prostate cancer PDX model

[0638] In the expression of PSMA ( Fig.24 A) In the androgen-dependent, patient-derived ST1273 model, a single intravenous injection of 75 or 150 kBq / kg of 225Ac-Pelgifatamab produced a high antitumor effect, with treatment / control (T / C) ratios of 0.22 and 0.05, respectively. Darolutamide monotherapy at 100 mg / kg (BID, oral) showed good antitumor effect, with a T / C ratio of 0.29 ( Fig.24 BC, Table 4).

[0639] Notably, the combination of 75 or 150 kBq / kg of 225Ac-Pelgifatamab with darolutamide (100 mg / kg) produced an additive antitumor effect, with a T / C ratio of 0.01 ( Fig.24 BC, Table 4). In addition, 5 / 10 and 6 / 10 mice were tumor-free 100 days after a single 225Ac-Pelgifatamab injection.

[0640] All treatments were well tolerated, as evidenced by stable body weight throughout the study period ( Fig.24D).

[0641] Table 4. 225 In vivo efficacy of Ac-Pelgifatamab and darolutamide in ST1273 and 22Rv1 prostate cancer models.

[0642]

[0643]

[0644] T / C, treatment / control ratio determined according to tumor volume.

[0645] Statistical analysis was performed using one-way ANOVA and Tukey's test.

[0646] **, p < 0.01 compared with vehicle; ***, p < 0.001 compared with vehicle.

[0647] # , p < 0.05 compared with darolutamide monotherapy; ### , p<0.001 compared with darolutamide monotherapy.

[0648] § , and the corresponding 225 Compared with Ac-Pelgifatamab monotherapy, p<0.05.

[0649] Darolutamide also enhances the antitumor efficacy of 225Ac-Pelgifatamab in the androgen-independent 22Rv1 prostate cancer model

[0650] In the expression of PSMA ( Fig.25 A) In the androgen-independent cell line-derived 22Rv1 model, a single intravenous injection of 150 kBq / kg of 225Ac-Pelgifatamab showed antitumor efficacy with a T / C ratio of 0.59 ( Fig.25 BC, Table 4). Darolutamide (100 mg / kg, oral, BID) monotherapy was ineffective, with a T / C ratio of 0.78 on day 16.

[0651] Notably, the combination of 225Ac-Pelgifatamab (150 kBq / kg) and darolutamide improved efficacy, with a T / C ratio of 0.20 ( Fig.25 BC, Table 4).

[0652] All treatments were well tolerated, as evidenced by stable body weight during the study ( Fig.25 D).

[0653] Flow cytometric analysis of isolated 22Rv1 tumors showed a 50-fold increase in PSMA expression after darolutamide treatment compared with vehicle ( Fig.25 E).

[0654] The androgen-independent prostate cancer model 22Rv1 was evaluated 225 In vivo antitumor efficacy of Ac-pelgi combined with darolutamide. Starting from day 15 after 22Rv1 tumor cell inoculation, male nude mice (n=10-12 mice / group, nude mice (nu / nu)) received a single injection of 225 Ac-pelgi (75 or 150 kBq / kg, intravenous injection, total protein dose 0.75 mg / kg) and / or darolutamide (100 mg / kg, BID, oral) were used for treatment. The anti-tumor effect of the combination treatment group was significantly improved compared with the two single treatment groups ( Fig. 22 ).

Claims

1. A tissue targeting compound of formula (I), wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for prostate-specific membrane antigen (PSMA).

2. The tissue targeting compound of formula (I) according to claim 1, wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, the anti-PSMA monoclonal antibody J591 or an antigen-binding fragment thereof, or the anti-PSMA monoclonal antibody TLX592 or an antigen-binding fragment thereof.

3. The tissue targeting compound of formula (I) according to claim 1, wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof.

4. A tissue targeting compound of formula (I) according to any one of claims 1 to 3, wherein [Ab] is the anti-PSMA antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.

10.

5. A tissue targeting compound of formula (I) according to claim 1 or 2, wherein [Ab] is the anti-PSMA antibody J591 or an antigen-binding fragment thereof, which comprises at least three CDR heavy chain sequences according to SEQ ID NO.13, SEQ ID NO.14 and SEQ ID NO.15 and three CDR light chain sequences according to SEQ ID NO.16, SEQ ID NO.17 and SEQ ID NO.

18.

6. A tissue targeting compound of formula (I) according to claim 1 or 2, wherein [Ab] is the anti-PSMA antibody TLX592 or an antigen-binding fragment thereof, which comprises at least three CDR heavy chain sequences according to SEQ ID NO.21, SEQ ID NO.22 and SEQ ID NO.23 and three CDR light chain sequences according to SEQ ID NO.24, SEQ ID NO.25 and SEQ ID NO.

26.

7. Method for preparing tissue targeting compounds of formula (I) wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for PSMA, The method comprises (i) coupling the chelating moiety of formula (II) to a monoclonal antibody or antigen-binding fragment thereof having binding affinity for PSMA, and (ii) contacting the resulting product with an aqueous solution comprising ions of at least one alpha-emitting actinium isotope.

8. The method according to claim 7, wherein [Ab] is the anti-PSMA antibody Pelgifatamab or an antigen-binding fragment thereof, The method comprises (i) coupling the chelating moiety of formula (II) to the anti-PSMA antibody Pelgifatamab or its antigen-binding fragment via a lysine residue of the monoclonal antibody or its antigen-binding fragment, and (ii) contacting the resulting product with an aqueous solution comprising ions of at least one alpha-emitting actinium isotope.

9. The method according to claim 7, wherein [Ab] is the anti-PSMA antibody Pelgifatamab or an antigen-binding fragment thereof, The method comprises (i) coupling the chelating moiety of formula (II) to the anti-PSMA antibody Pelgifatamab or its antigen-binding fragment via a lysine residue of the monoclonal antibody or its antigen-binding fragment, and (ii) contacting the resulting product with an aqueous solution comprising ions of at least one alpha-emitting actinium isotope, The anti-PSMA antibody Pelgifatamab or its antigen-binding fragment comprises at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.

10.

10. A tissue targeting chelator of formula (III), Wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof that has binding affinity for PSMA.

11. The tissue targeting chelator of formula (III) according to claim 10, wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, the anti-PSMA monoclonal antibody J591 or an antigen-binding fragment thereof, or the anti-PSMA monoclonal antibody TLX592 or an antigen-binding fragment thereof.

12. The tissue targeting chelator of formula (III) according to claim 10, wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof.

13. A tissue targeting chelator of formula (III) according to any one of claims 10 to 12, wherein [Ab] is the anti-PSMA antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.

10.

14. A tissue targeting chelator of formula (III) according to claim 10 or 11, wherein [Ab] is the anti-PSMA antibody J591 or an antigen-binding fragment thereof, which comprises at least three CDR heavy chain sequences according to SEQ ID NO.13, SEQ ID NO.14 and SEQ ID NO.15 and three CDR light chain sequences according to SEQ ID NO.16, SEQ ID NO.17 and SEQ ID NO.

18.

15. A tissue targeting chelator of formula (III) according to claim 10 or 11, wherein [Ab] is the anti-PSMA antibody TLX592 or an antigen-binding fragment thereof, which comprises at least three CDR heavy chain sequences according to SEQ ID NO. 21, SEQ ID NO. 22 and SEQ ID NO. 23 and three CDR light chain sequences according to SEQ ID NO. 24, SEQ ID NO. 25 and SEQ ID NO.

26.

16. Use of a tissue targeting chelator of formula (III) according to any one of claims 10 to 15 for the preparation of a tissue targeting compound of formula (I) according to any one of claims 1 to 6.

17. A combination comprising (i) Tissue-targeting compounds of formula (I) wherein [Ab] is a monoclonal antibody or an antigen-binding fragment thereof having binding affinity for prostate-specific membrane antigen (PSMA), and (ii) an agent selected from the group consisting of nonsteroidal antiandrogens, steroidal antiandrogens, androgen synthesis inhibitors, antigonadotropins, PARP inhibitors, ATR inhibitors, ATM inhibitors, DNA-PK inhibitors, AKT inhibitors, Pi3K inhibitors, PSMA-targeted beta emitters, immune checkpoint inhibitors, alpha emitters, vaccines, and chemotherapeutic agents.

18. The combination according to claim 17, wherein [Ab] is the anti-PSMA monoclonal antibody Pelgifatamab or an antigen-binding fragment thereof, which comprises at least three CDR heavy chain sequences according to SEQ ID NO.5, SEQ ID NO.6 and SEQ ID NO.7 and three CDR light chain sequences according to SEQ ID NO.8, SEQ ID NO.9 and SEQ ID NO.

10.

19. A combination according to claim 17 or 18, wherein the agent is selected from the group consisting of nonsteroidal antiandrogens, steroidal antiandrogens, androgen synthesis inhibitors, antigonadotropins, PARP inhibitors, PSMA-targeted beta emitters, alpha emitters, vaccines and chemotherapeutic agents.

20. A combination according to any one of claims 17 to 19, wherein the agent is selected from nonsteroidal antiandrogens, steroidal antiandrogens, androgen synthesis inhibitors and antigonadotropins.

21. A combination according to any one of claims 17 to 20, wherein the agent is selected from the group consisting of nonsteroidal antiandrogens, steroidal antiandrogens, androgen synthesis inhibitors, antigonadotropins and PARP inhibitors, wherein the nonsteroidal antiandrogen is selected from the group consisting of darolutamide, bicalutamide, enzalutamide, apalutamide, flutamide, nilutamide and topiramide, and the steroidal antiandrogen is selected from the group consisting of cyproterone acetate, allylestradiol, chlormadinone acetate, delmadinone acetate, progesterone caproate, hydroxyprogesterone caproate, medroxyprogesterone acetate, hydroxy ... The invention relates to a pharmaceutical composition comprising the following: progesterone, megestrol acetate, osaterol acetate, oxentolone and spironolactone, the androgen synthesis inhibitor is selected from ketoconazole, abiraterone, aminoglutethimide, goserelin and severonil, the antigonadotropin is selected from abarelix, danazol, gestrinone, parahydroxypropiophenone, cetrorelix, degarelix, alagrogol sodium, ganirelix, linzagoli and relugolix, and the PARP inhibitor is selected from olaparib, rucaparib, veliparib, niraparib, talazoparib, pamiparib, CEP 9722, E7016 and iniparib.

22. A combination according to any one of claims 17 to 21, wherein the agent is selected from the group consisting of nonsteroidal antiandrogens, steroidal antiandrogens, androgen synthesis inhibitors and antigonadotropins, wherein the nonsteroidal antiandrogen is selected from the group consisting of darolutamide, bicalutamide, enzalutamide, apalutamide, flutamide, nilutamide and topiramide, and the steroidal antiandrogen is selected from the group consisting of cyproterone acetate, allylestradiol, chlormadinone acetate, delmagestrol acetate, The invention relates to a method for preparing an oxytocin-containing drug and a preparation for preparing the oxytocin-containing drug. The method comprises the following steps: the first step is to prepare an oxytocin-containing drug and a preparation for preparing the oxytocin-containing drug. The first step is to prepare an oxytocin-containing drug and a preparation for preparing the oxytocin-containing drug. The second step is to prepare an oxytocin-containing drug and a preparation for preparing the oxytocin-containing drug.

23. A combination according to any one of claims 17 to 22, wherein the agent is selected from the nonsteroidal antiandrogen darolutamide, the steroidal antiandrogen cyproterone acetate, the androgen synthesis inhibitors abiraterone or goserelin and the antigonadotropins degarelix or relugolix.

24. A combination according to any one of claims 17 to 23, wherein the agent is the non-steroidal antiandrogen darolutamide.

25. A tissue targeting compound of formula (I) according to any one of claims 1 to 6 or a combination according to any one of claims 17 to 24 for use in the treatment of a disease.

26. A tissue targeted compound of formula (I) according to any one of claims 1 to 6 or a combination according to any one of claims 17 to 24 for use according to claim 25, wherein the disease is characterized by overexpression of prostate specific membrane antigen (PSMA).

27. A tissue targeting compound of formula (I) according to any one of claims 1 to 6 or a combination according to any one of claims 17 to 24 for use according to claim 25 or 26, wherein the disease is cancer, preferably prostate cancer.

28. A tissue targeting compound of formula (I) according to any one of claims 1 to 6 or a combination according to any one of claims 17 to 24 for use in the treatment of a disease, preferably wherein the disease is a hyperproliferative disease, more preferably wherein the hyperproliferative disease is cancer.

29. Use of a tissue targeting compound of formula (I) according to any one of claims 1 to 6 or a combination according to any one of claims 17 to 24 for treating a disease, preferably wherein the disease is a hyperproliferative disease, more preferably wherein the hyperproliferative disease is cancer.

30. Use of a tissue targeting compound of formula (I) according to any one of claims 1 to 6 or a combination according to any one of claims 17 to 24 for the preparation of a medicament for treating a disease, preferably wherein the disease is a hyperproliferative disease, more preferably wherein the hyperproliferative disease is cancer.

31. A method of treating a disease, preferably wherein the disease is a hyperproliferative disease, more preferably wherein the hyperproliferative disease is cancer, the method comprising administering to the subject a therapeutically effective amount of a tissue targeted compound of formula (I) according to any one of claims 1 to 6 or a combination according to any one of claims 17 to 24.

32. A tissue targeted compound, combination, use or method according to any one of claims 25 to 31 , wherein the disease is prostate cancer.

33. A composition comprising a combination according to any one of claims 17 to 24 and a pharmaceutically acceptable ingredient.

34. A kit comprising the following combination Tissue targeting compounds and agents of formula (I) according to any one of claims 1 to 6; and optionally one or more other agents; Optionally, both or one of the tissue targeting compound or the agent of formula (I) is in the form of a pharmaceutical preparation, which can be administered simultaneously, concurrently, separately or sequentially.

Images