Porphyrins and their pharmaceutical uses

Abstract

The present invention relates to a method for preparing a porphyrin compound of formula (I) or its metallo-derivatives, salts, and solvates, particularly its tin(IV) phosphate derivative [wherein R1, R2, R3, R4, R5, and R6 are each independently H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 The present invention relates to porphyrins selected from alkanamines and C(O)CH3. The present invention also relates to pharmaceuticals, in particular pharmaceuticals for use in the treatment of proliferative and / or malignant diseases (such as cancer) and for use in treating or inhibiting metastasis, as well as pharmaceutical compositions containing such porphyrins. [Formula 1] JPEG2026509438000119.jpg72170

Description

[Technical Field]

[0001] The present invention relates to methods for preparing porphyrins or their metallo-derivatives, salts, and solvates, particularly methods for preparing staghorn mesoporphyrins, and compounds prepared by such methods. The present invention also relates to such porphyrins or their metallo-derivatives, salts, and solvates for use as pharmaceuticals, particularly for the treatment of proliferative and / or malignant diseases (such as cancer), and to pharmaceutical compositions containing such porphyrins. [Background technology]

[0002] Cancer is a leading cause of death worldwide. The need for new cancer treatments to address the disease remains urgent.

[0003] One promising treatment involves immunotherapy that targets specific proteins. Preclinical data demonstrate that heme oxygenase-1 (HO-1) represents an important immunotherapy target in cancer (Non-Patent Literature 1 (see Reference 5 below)).

[0004] The HO family of enzymes includes the biologically active product of heme, biliverdin, and ferrous (Fe) 2+ ), and is responsible for its decomposition to carbon monoxide (CO) (Non-Patent Literature 2 (see Reference 6 below)). CO has shown immunosuppressive properties (Non-Patent Literature 3 (see Reference 7 below), Non-Patent Literature 4 (see Reference 8 below), Non-Patent Literature 5 (see Reference 9 below)), suggesting its potential role in blocking the immune response against tumors. Using a high-grade spontaneous breast cancer model (MMTV-PyMT), the inventors demonstrated that the combination of sparrow mesoporphyrin (SnMP), a clinically important small molecule inhibitor of HO-1, with standard chemotherapy reduced immunosuppression and allowed CD8+ T cells (immune cells that can specifically target cancer) to control tumor growth (Non-Patent Literature 1, Non-Patent Literature 6 (see Reference 10 below)).

[0005] SnMP is a tin adduct of mesoporphyrin IX (3,3'-(7,12-diethyl-3,8,13,17-tetramethylporphyrin-2,18-diyl)dipropionic acid), as shown in Scheme 1 below. KCl-HO-1i is the tin phosphate salt of mesoporphyrin IX, and its synthesis is shown in Scheme 2 below. KCl-HO-1i is an HO inhibitor as shown in the example below. SnMP and KCl-HO-1i cross-react with HO-1 / 2.

[0006] Conventional synthesis of SnMPs involves the use of hemin (usually derived from bovine sources), which can raise ethical, material sourcing, and hygiene concerns, and has raised regulatory issues in several markets.

[0007] SnMP as a chloride salt is described in the literature as an HO-1 and HO-2 inhibitor. SnMP has shown tumor suppression when used in combination with existing chemotherapeutic or immunotherapeutic agents. Our research suggests that the combination of SnMP and standard chemotherapy can play a role in controlling tumor growth by NK cells.

[0008] Infant hyperbilirubinemia (also known as infantile jaundice or neonatal hyperbilirubinemia) occurs in newborns when the liver is unable to conjugate bilirubin in a way that keeps pace with its formation and excretion rate. Bilirubin is produced at birth from the release of heme, which is part of the physiological conversion of fetal hemoglobin to adult hemoglobin. Stansoporfin (tin(IV) mesoporphyrin IX dichloride) has been shown to have therapeutic value in the treatment of hyperbilirubinemia. [ka] Scheme 1: Mesoporphyrin IX

[0009] The synthesis and use of SnMP are described.

[0010] International Publication No. 2013 / 083659 discloses cancer treatments including combinations of HO-1 inhibitors such as SnMPs and immunotherapies.

[0011] Japanese Patent Application No. 2013 / 232129 discloses a method for synthesizing stanzaporfin (tin(IV) mesoporphyrin IX dichloride) in high purity and on a large scale using hemin as a starting material, and the resulting composition.

[0012] U.S. Patent No. 10,533,024 discloses a method for synthesizing metal mesoporphyrins by transmetallation of hemin followed by hydrogenation of tin protoporphyrin IX to form metal mesoporphyrins.

[0013] U.S. Patent No. 8,530,458 discloses a method for large-scale, high-purity synthesis of stansoporfin (tin(IV)mesoporphyrin IX dichloride), the compositions thus produced, and their use for the treatment of infantile hyperbilirubinemia. [Prior art documents] [Patent Documents]

[0014] [Patent Document 1] International Publication No. 2013 / 083659 [Patent Document 2] Patent application No. 2013 / 232129 [Patent Document 3] U.S. Patent No. 10,533,024 [Patent Document 4] U.S. Patent No. 8,530,458 [Non-patent literature]

[0015] [Non-Patent Document 1] T. Muliaditan et al., Repurposing tin mesoporphyrin as an immune checkpoint inhibitor shows therapeutic efficacy in preclinical models of cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, (2018) [Non-Patent Document 2] R. Gozzelino et al., Mechanisms of cell protection by heme oxygenase-1. Annual review of pharmacology and toxicology 50, 323-354 (2010) [Non-Patent Document 3] K. Minamoto et al., Reciprocal regulation of airway rejection by the inducible gas-forming enzymes heme oxygenase and nitric oxide synthase. The Journal of experimental medicine 202, 283-294 (2005) [Non-Patent Document 4] S. Brouard et al., Carbon monoxide generated by heme oxygenase 1 suppresses endothelial cell apoptosis. The Journal of experimental medicine 192, 1015-1026 (2000) [Non-Patent Document 5] X. Zhang et al., Carbon monoxide differentially modulates STAT1 and STAT3 and inhibits apoptosis via a phosphatidylinositol 3-kinase / Akt and p38 kinase-dependent STAT3 pathway during anoxia-reoxygenation injury. The Journal of biological chemistry 280, 8714-8721 (2005) [Non-Patent Document 6] JN Arnold et al., Tumoral immune suppression by macrophages expressing fibroblast activation protein-alpha and heme oxygenase-1. Cancer immunology research 2, 121-126 (2014) [Overview of the project] [Problems that the invention aims to solve]

[0016] However, there is a need for improved methods for producing mesoporphyrins and their derivatives, particularly synthetic methods that do not rely on animal products (such as hemins). Improved therapeutic forms of mesoporphyrins and their derivatives, including new salt forms, are also desired.

[0017] The inventors have found that several tin salts of mesoporphyrin derivatives can provide improved treatment for proliferative and / or malignant diseases and can also inhibit or reduce metastasis.

[0018] This invention aims to address these needs and overcome the problems associated with the prior art. [Means for solving the problem]

[0019] In view of the above, the present invention, in a first aspect, relates to a compound of formula (I): [ka] Or a method for preparing a metallo-dera, salt, and solvate thereof, comprising the dicarboxylic acid compound of the following formula: [ka] Or its salts and solvates, the dialdehyde compounds of the following formula: [ka] Alternatively, it can be added to its salts and solvates and reacted to form the following porphyrin diester compound: [ka] or the step of forming a metallo-derivative, salt, and solvate thereof. [In the formula, R1, R2, R3, R4, R5, and R6 are each independently H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', and C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3, R7 and R8 are independently C 1-6 Selected from alkyl, R' and R'' are independently H and C 1-6 The present invention provides a method that includes [selected from alkyl groups].

[0020] In a further embodiment, a compound of formula (I) or its metallo-derivatives, salts, and solvates, wherein the compound is [ka] The present invention provides compounds or their metallo-derivatives, salts, and solvates, which are salts and solvates thereof.

[0021] More precisely, the compound of formula (II) or (III) from the following formulas: [ka] Alternatively, salts and solvates thereof are provided.

[0022] More precisely, the compound of formula (II) or (III) is such that R1, R3, R5, and R6 are methyl, and R2 and R4 are ethyl, and as a result, the compound of formula (VI) or (VII): [ka] Or it becomes a salt or solvate thereof.

[0023] In a further embodiment, compounds of formula (II), (III), (IV), (V), or (VI), or salts and solvates thereof, are provided for use as pharmaceuticals.

[0024] In a further embodiment, compounds of formula (II), (III), (IV), (V), or (VI), or salts and solvates thereof, are provided for use in the treatment of proliferative and / or malignant diseases.

[0025] In a further embodiment, compounds of formula (II), (III), (IV), (V), or (VI), or salts and solvates thereof, are provided for use in therapeutic methods.

[0026] In a further embodiment, the present invention provides a pharmaceutical composition comprising a compound of formula (II), (III), (IV), (V), or (VI), or a salt or solvate thereof, and a pharmaceutically acceptable excipient, carrier, or diluent.

[0027] In a further embodiment, compounds of formula (II), (III), (IV), (V), or (VI), or salts and solvates thereof, are provided for use in the manufacture of pharmaceuticals for the treatment of proliferative and / or malignant diseases.

[0028] In a further aspect, there is provided a method of treating a patient suffering from a proliferative and / or malignant disease, the method comprising administering to the patient a therapeutically effective amount of a compound of formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof.

[0029] In a further aspect, there is provided a compound of formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof, wherein the compound is administered alone or co-administered or sequentially administered with one or more immunotherapeutic agents or other anti-cancer agents for use in the treatment of proliferative and / or malignant diseases.

[0030] In a further aspect, a compound of formula (I):

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[0031] In a further embodiment, compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, or mixtures thereof, may be administered alone, concurrently, or sequentially in combination with other therapeutic agents, depending on the condition being treated.

[0032] In a further embodiment, compounds of formula (II), (III), (IV), (V), or (VI), or salts and solvates thereof, are provided for use in the treatment or inhibition of metastasis. [Brief explanation of the drawing]

[0033] Next, embodiments of the present invention will be further described with reference to the attached drawings. [Figure 1] This graph shows the serum drug concentration (ng / mL) in mouse plasma samples after administration of SnMP (38 mg / kg, PO), analyzed by LC-MS / MS. The mean ± standard deviation versus time is plotted as a line graph (semi-logarithmic scale). [Figure 2] This graph shows the serum drug concentration (ng / mL) in mouse plasma samples after administration of KCL-HO-1i(NS) (44 mg / kg, PO), analyzed by LC-MS / MS. The mean ± standard deviation versus time is plotted as a line graph (semi-logarithmic scale). [Figure 3] This graph combines the data from Figures 1 and 2. [Figure 4A] ~ [Figure 4B] This is a schematic diagram illustrating intraperitoneal (ip) administration strategies for KCL-HO-1i or SnMP and / or fluorouracil (5-FU) or gemcitabine or a vehicle in MMTV-PyMT mice with established tumors. [Figure 4C] ~ [Figure 4G]Figures 4A and 4B show growth curves of established spontaneous tumors in MMTV-PyMT mice treated with a single medium, SnMP, KCL-HO-1i, 5-FU, or gemcitabine, according to the administration strategies indicated. Each line represents an individual tumor and mouse. [Figure 4H] ~ [Figure 4J] Figures 4A and 4B show the growth curves of established spontaneous tumors in MMTV-PyMT mice treated with combination therapy of SnMP / 5-FU, KCL-HO-1i / 5-FU, and KCL-HO-1i / gemcitabine, as instructed by the administration strategies shown. Each line represents an individual tumor and mouse. [Figure 4K] This figure shows the average growth curves of established spontaneous tumors in mice given the indicated medium, KCL-HO-1i, 5-FU, or a combination of KCL-HO-1i and 5-FU. [Figure 4L] This figure shows the mean growth curves of established spontaneous tumors in mice given the indicated medium, KCL-HO-1i, gemcitabine, or a combination of KCL-HO-1i and gemcitabine. [Figure 5] This figure shows the tumor size 8 days after the start of treatment in MMTV-PyMT mice treated with SnMP, KCL-HO-1i, along with fluorouracil (5-FU). Each dot represents an individual mouse. * P<0.05. [Figure 6] This figure shows the growth curves of spontaneously occurring tumors that encroached in MMTV-PyMT mice treated with the prescribed commercially available SnMP, SnMP synthesized according to the present invention (vegan SnMP), and / or fluorouracil (5-FU). Each line represents an individual mouse. [Figure 7A] This is a schematic diagram showing the experimental and explanatory (representation) of the strategy used to determine plasma concentrations. [Figure 7B] This graph shows the time course of plasma concentrations in male mice that received commercially available SnMP, SnMP synthesized according to the present invention (veSnMP), or KCL-HO1i via intraperitoneal administration (left panel) or oral administration (right panel). [Figure 7C] The HO-1luc / eGFP reporter mouse model used (top) and a brief description of the experiment (bottom) are shown. [Figure 7D] This figure shows descriptive bioluminescence images of HO-1luc / eGFP reporter mice (C) at whole-body T0 (upper panel) and 24 hours after treatment (lower panel) for media, commercially available SnMP, SnMP synthesized according to the present invention (VeSnMP), or KCL-HO1i. [Figure 7E] This figure shows the relative quantification of luciferase expression (photons / second) in the whole-body image shown in (D) of the HO-1luc / eGFP reporter mouse (C). [Figure 7F] This figure shows the changes in HO-1 expression in individual mouse tissues (derived from those shown in D) normalized to the media treatment group in HO-1luc / eGFP reporter mice (C). [Figure 7G] This figure shows the evaluation of inhibition of HO-1 activity in rat microsomes using various doses of KCL-HO-1i and SnMP. [Figure 8A] This is a schematic diagram illustrating the administration strategy for KCL-HO-1i and gemcitabine in MMTV-PyMT mice. [Figure 8B] This figure shows the change in tumor volume (mm3) in each mouse against time (days). KCL-HO-1i was delivered orally, and gemcitabine was delivered intraperitoneally. The dashed line indicates the start of treatment. [Figure 9] This bar graph shows CD8 T cell infiltration into tumors of MMTV-PyMT mice treated with a medium, commercially available SnMP (cSnMP), or KCL-HO-1i. [Figure 10A] This is a schematic diagram illustrating the intraperitoneal administration strategy for non-immune IgG and immunodepleted anti-CD8α antibodies administered together with KCL-HO-1i and gemcitabine in MMTV-PyMT mice. [Figure 10B] This figure shows the growth curves of individual mice treated with non-immune IgG. [Figure 10C]This figure shows the growth curves of individual mice treated with immunodepletion anti-CD8α antibodies. [Figure 10D] This figure shows the growth curves of individual mice treated with immunodepletion anti-CD8α antibody, KCL-HO-1i, and gemcitabine, respectively. [Figure 10E] This figure shows growth curves, where each line represents the average for each treatment. [Figure 11A] This is a schematic diagram illustrating the intraperitoneal administration strategy of KCL-HO-1i and / or 5-FU or gemcitabine or a medium in MMTV-PyMT mice with established tumors. The tumors were analyzed by flow cytometry 36 hours after the start of treatment. [Figure 11B] This figure shows the changes in tumor growth over a 36-hour treatment period. [Figure 11C] This figure shows the stromal composition of enzyme-dispersed tumors for each of the different treatments. [Figure 11D] These bar graphs represent CD8+ T cell infiltration in the tumor microenvironment for each of the different treatments. [Figure 11E] This bar graph shows the CD8+ T cell effector function, as evaluated by IFN-γ expression, for each of the different treatments. [Figure 11F] This bar graph represents CD8+ T cell infiltration in the tumor microenvironment in relation to the medium and SnMPs. [Figure 11G] This figure shows representative dot plots of FACS-gated (7AAD-), CD45+CD3+CD8+ T cells showing CD44 and CD62L expression in media-treated mice. [Figure 11H] This is a histogram representing the CD8+ T cell subpopulation (shown in Figure 11G) across different treatment groups. [Figure 12A] Figure 12A is a schematic diagram showing the administration strategy to mice with encroaching MMTV-PyMT tumors treated with KCL-HO-1i and / or 5-FU or gemcitabine or a medium, with tumor tissue analyzed at 36 hours by bulk RNA-seq analysis. [Figure 12B]Figure 12B is a Venn diagram showing all DEGs for each treatment of the medium and the intersections between these groups. [Figure 12C] Figure 12C is a Venn diagram showing all the up-control DEGs for KCL-HO-1i treatment of the medium and the crossovers between these groups. [Figure 12D] Figure 12D shows a heatmap of common upregulatory DEGs (419 genes) that have been hierarchically clustered across treatment groups, which are secretory genes (91 genes) throughout the entire treatment process. [Figure 12E] Figure 12E is a Venn diagram showing all upcontrol DEGs for treatments including chemotherapy and the crossovers between these groups. [Figure 12F] Figure 12F is a Venn diagram showing dual therapy treatments associated with chemokines and upregulated DEG, as well as crossovers between these groups. [Figure 12G] Figure 12G is a heatmap showing hierarchical clustering of chemokine and cytokine upregulation DEGs associated with the dual-drug treatment group. [Figure 13A] This is a schematic diagram illustrating the administration strategy of KCL-HO-1i and / or gemcitabine or a vehicle in MMTV-PyMT mice with established tumors. [Figure 13B] Figure 13A shows the tumor growth curves for each treatment. [Figure 13C] This graph shows the weight of the mice during each treatment. [Figure 13D] This figure shows the ratio of serum liver aspartate aminotransferase (AST) to alanine aminotransferase (ALT) in the blood for each treatment. [Figure 13E] This diagram shows the blood immune cell composition for each treatment. [Figure 13F] This figure shows a heatmap of the physical characteristics of the animals obtained on the 21st day after treatment. [Figure 14A]This is a schematic diagram illustrating the oral delivery strategy for KCL-HO-1i and / or ip-delivered gemcitabine or a medium in C57Bl / 6 mice (MN-MCA-1 cell line) with established subcutaneous sarcoma tumors. [Figure 14B] Figure 14A shows the tumor growth curves for each treatment. [Figure 15A] This figure shows the tumor volume of mice that received KCL-HO-1i (blue curve) or the medium (black curve) daily for 14 days, 12 days after tumor injection. [Figure 15B] This histogram shows the counted metastatic nodules from lung samples taken 26 days after tumor injection. [Modes for carrying out the invention]

[0034] definition The following abbreviations are used throughout this specification: Ac Acetyl-C(O)CH3; Et Ethyl; E3N Triethylamine; 5-FU Fluorouracil; KCl-HO-1i Tin phosphate of mesoporphyrin IX; Me Methyl; MeOH Methanol; Ph (Phenyl); p-TSA / PTSA p-toluenesulfonic acid; TBAC Tetrabutylammonium chloride; TEOF Triethyl orthoformic acid; TFA Trifluoroacetic acid; THF Tetrahydrofuran; Ts(Tosylate) p-toluenesulfonyl group [-SO2-C6H4-CH3]; ip Intraperitoneal administration; SD Standard deviation; SEM Standard error of the mean.

[0035] Adoptive cell therapy is an immunotherapy that uses the immune cells of the subject (or donor immune cells) to treat diseases such as cancer or viral infections. In adoptive cell therapy, T cells are isolated based on their ability to proliferate in response to tumors, or genetically modified to target certain molecules on cells, such as antigens on cancer cells. The tumor-responsive T cells are then expanded and injected back into the subject.

[0036] TIL therapy is a form of adoptive cell therapy involving tumor-infiltrating lymphocytes. In TIL therapy, tumor-infiltrating lymphocytes that have penetrated the tumor are collected from tumor biopsy material taken from the patient, grown in vitro, and reinjected into the patient. Tumor-infiltrating lymphocytes actively participate in the destruction of the tumor. In one method, following the excision of the biopsy material, DNA isolated from the tumor is sequenced to identify cancer-specific mutations that are recognized as neoantigens. In this method, the mutated neoantigens are inserted into autologous dendritic cells and co-cultured with tumor-infiltrating lymphocytes. The tumor-infiltrating lymphocytes are then assayed for neoantigen recognition. These tumor-infiltrating lymphocytes that recognize the neoantigens are then selected, grown, and transfused back into the patient. In another method, a number of T cells are grown due to the ability of T cells to recognize isolated tumor biopsy material, and then injected back into the patient.

[0037] Another approach to adoptive cell therapy is called TCR therapy, which involves modifying the T cells of a subject or donor to express a specific T cell receptor ("TCR"). The T cell receptor is a heterodimer consisting of two subunits, TCRα and TCRβ. Each subunit contains a constant region that tethers the receptor to the cell membrane and a hypervariable region that functions in antigen recognition. TCRs can recognize tumor-specific proteins / peptides presented on the extracellular surface. In TCR therapy, T cells are collected from the blood of the subject or donor. The T cells are genetically modified in the laboratory to express a new T cell receptor. The number of T cells is increased and injected back into the subject. T cells with the new T cell receptor can target the patient's cancer.

[0038] A further form of adoptive cell therapy is chimeric antigen receptor ("CAR") T cell therapy ("CAR-T therapy"). In CAR-T therapy, one or more portions of the T cell receptor are modified into antigen-binding portions such as antibodies, antibody fragments, or receptor ligands. Cancer-associated antigens (tumor-associated antigens or "TAAs") are often expressed by tumors. Antibodies or antibody fragments target TAAs. T cells that target TAAs can directly attack cancer cells. In CAR-T therapy, T cells are collected from the blood of the patient or donor, genetically modified to express CARs, and then the number of T cells is increased and injected back into the patient. CAR-T modification allows T cells to specifically target the target cancer.

[0039] When "substituted" is used in relation to a chemical substituent or moiety (e.g., an alkyl group), it means that one or more hydrogen atoms of the substituent or moiety are replaced by one or more nonhydrogen atoms or groups, provided that the valence requirements are met and the compound is chemically stable as a result of the substitution.

[0040] The phrases "independently" or "independently selected" are used in the context of statements such as "R1 and R2 are independently H, methyl, ethyl, ..." and mean that each example of a functional group, e.g., R1, is selected from the enumerated options, separate from any other examples of R1 or R2 in the compound. For example, H can be selected as the first example of R1 in the compound, methyl can be selected as the second example of R1 in the compound, and ethyl can be selected as the first example of R2 in the compound.

[0041] C 1-6 Alkyl groups are generally linear and branched saturated hydrocarbon groups having 1 to 6 carbon atoms; appropriately, C 1-5 Alkyl; more appropriately C 1-4 Alkyl; more appropriately C 1-3Alkyl; more appropriately, refers to methyl or ethyl. Examples of alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl, penta-1-yl, penta-2-yl, penta-3-yl, 3-methylbuta-1-yl, 3-methylbuta-2-yl, 2-methylbuta-2-yl, 2,2,2-trimethylethyl-1-yl, n-hexyl, n-heptyl, and n-octyl.

[0042] C 1-2 Alkanamines refer to methyl or ethyl groups of amines substituted with the NR'R'' group, where R' and R'' are independently H and C, respectively. 1-6 Selected from alkyl groups. Therefore, C 1-2 Alkanamines are CH2NR'R'', CHNR''R''-CH3, or CH2-CH2NR'R''.

[0043] C 1-2 Haloalkyl refers to a methyl or ethyl group substituted with one or more halogen groups. Therefore, C 1-2 Haloalkyls are CH2X, CHX2, CX3, CH2CH2X, CH2CHX2, CH2CX3, CHXCH3, CHXCH2X, CHXCHX2, CHXCX3, CX2CH3, CX2CH2X, CX2CHX2, or CX2CX3, where X is a halogen. More precisely, C 1-2 The haloalkyl group is CH2X, CX3, CH2CH2X, CHXCH3, or CX2CX3. Each halogen can be selected independently. More appropriately, C 1-2 Each halogen in the haloalkyl group is the same.

[0044] "Halo," "halogen," and "halogeno" can be used synonymously, and halogens are each independently selected from fluoro, chloro, bromo, and iodine. More appropriately, halogens are each independently fluoro or chloro.

[0045] The term "or its metallo derivatives, salts, and solvates" means that the porphyrin compound may, as an alternative, not contain any metallo group, salt, or solvate, or may contain one or more of them. Thus, the compound may be a metallo derivative, a salt, or a solvate, or any combination thereof. A metallo derivative is a compound in which a metal is housed at the center of the porphyrin ring. The most preferred metallo group is tin. For example, the compounds of formula (II) or (III) described herein are tin metallo derivatives of the compound of formula (I), and also include phosphates.

[0046] The term "oncolytic virus" is used in its simple, common sense to refer to a virus that preferentially infects and kills cancer cells. When infected cancer cells are destroyed by tumor breakdown (direct lysis of cells), they release new infectious viral particles or virions, which help destroy the remaining tumor. Oncolytic viruses are thought to not only cause the direct destruction of tumor cells but also stimulate the host's anti-tumor immune system response. Appropriate oncolytic viruses include adenoviruses, reoviruses, measles, herpes simplex virus, Newcastle disease virus, vaccinia, or seneca virus.

[0047] The term "or pharmaceutically acceptable salts, solvates, tautomers, stereoisomers, or mixtures thereof" means that pharmaceutically acceptable salts, solvates, tautomers, and stereoisomers of the indicated structure are also included. "Mixtures thereof" means that mixtures of these forms may exist; for example, the compounds of the present invention may contain both tautomers and pharmaceutically acceptable salts. A "pharmaceutically acceptable" substance is one that is appropriate for use in contact with the target tissue without excessive toxicity, irritation, or allergic response, corresponds to a reasonable benefit-risk ratio, is effective for its intended use, and falls within reasonable medical judgment.

[0048] A "pharmaceutical composition" refers to a combination of one or more active pharmaceutical ingredients and one or more excipients.

[0049] The term "or its salts and solvates" means that the compound may not contain any salts or solvates, or it may contain one or more. Therefore, the compound may be a salt, a solvate, or both. Appropriately, the broader term "or its metallo-derivatives, salts, and solvates" can be limited to "or its salts and solvates." If the compound contains salts, there may be one or more salts; for example, there may be situations where only one of the carboxylic acid groups is a salt, or where both carboxylic acid groups are salts. Preferably, any salt is a pharmaceutically acceptable salt.

[0050] In this specification, “solvate” refers to a variable stoichiometric complex formed by a solute and a solvent. For crystalline compounds in which solvent molecules are incorporated into the crystal lattice during crystallization, pharmaceutically acceptable solvates can be formed. The incorporated solvent molecules may be water molecules, or non-aqueous molecules such as ethanol, isopropanol, dimethyl sulfoxide, acetic acid, ethanolamine, and ethyl acetate molecules, but are not limited to these.

[0051] "Tautomers" refer to two or more isomers of a compound that exist together in equilibrium and readily interconvert through the transfer of atoms, groups, or double bonds within the molecule.

[0052] In this specification, the term “subject” refers to human or non-human mammals. Examples of non-human mammals include domesticated animals such as sheep, horses, cows, pigs, goats, rabbits, and deer; as well as companion animals such as cats, dogs, rodents, and horses. Preferably, the subject is human.

[0053] A "therapeutic dose" of a drug refers to the amount of a drug or composition that is effective in treating a subject and thus producing the desired therapeutic, ameliorative, inhibitory, or preventive effect. The therapeutic dose may depend, in particular, on the subject's weight and age, as well as the route of administration.

[0054] "To treat" means to reverse, reduce, or inhibit the progression of a disorder, disease, or condition to which such term applies, or to prevent a disorder, disease, or condition, or to reverse, reduce, or inhibit the progression of one or more symptoms of such disorder, disease, or condition, or to prevent one or more symptoms of such disorder, disease, or condition.

[0055] "Treatment" refers to the act of "providing treatment," as defined directly above.

[0056] In this specification, the term “comprising” means “including at least a portion of,” and is comprehensive or open-ended. When interpreting each statement in this specification that contains the term “comprising,” there may be features, elements and / or steps other than those preceded by the term. Related terms such as “comprise” and “comprises” should be interpreted in the same way.

[0057] The phrase "essentially derived from" limits the scope of the claim to the specified material or step of the claimed invention, "and which does not substantially affect the basic and novel features." When the phrase "essentially derived from" appears in a clause of the body of the claim rather than immediately after the preamble, it limits the scope to only the elements described in that clause.

[0058] The term "consisting of" excludes any element, step, or material not specified in the claim. "Consisting of" is defined as closing the claim to any material other than the material described, except for impurities that are normally associated with it. When the phrase "consisting of" appears in a section of the body of the claim rather than immediately following the preamble, it limits only the elements described in that section. Other elements are not excluded from the claim as a whole. It should also be understood that various embodiments in this specification are presented using the phrase "including" in various contexts, and related embodiments are described using the phrases "essentially consisting of" or "consisting of".

[0059] Deprotection reaction Appropriately, the method involves the following diester compound: [ka] Alternatively, by subjecting its salt and solvate to a deprotection reaction, the following dicarboxylic acid compound is obtained: [ka] The process further includes the step of forming a salt or solvate thereof.

[0060] More precisely, the diester compound of the following formula: [ka] Alternatively, by subjecting its salt and solvate to a deprotection reaction, the following dicarboxylic acid compound is obtained: [ka] or a step of forming a salt or solvate thereof.

[0061] Suitablely, a dicarboxylic acid compound is formed by subjecting a diester compound or its salt and solvate to a deprotection reaction using (i) tin and concentrated HCl; (ii) iron and ammonium formate; (iii) CrO3 and acetic acid; (iv) N-bromosuccinimide (NBS) and ozone; or (v) a hydrogenation reaction.

[0062] Ideally, the deprotection reaction is carried out by refluxing the diester compound or its salt and solvate with (i) tin and concentrated HCl.

[0063] Ideally, the deprotection reaction is carried out using ethanol and iron(II) in water and ammonium formate.

[0064] Ideally, the deprotection reaction is carried out using (iii)CrO3 and acetic acid.

[0065] Ideally, the deprotection reaction is carried out using (iv)N-bromosuccinimide (NBS) and ozone.

[0066] Appropriately, the deprotection reaction is (v) a hydrogenation reaction. Appropriately, the hydrogenation reaction is carried out using a Pd / C catalyst.

[0067] Ideally, the hydrogenation reaction should be carried out in the presence of an organic base.

[0068] Ideally, hydrogenation is carried out in the presence of triethylamine.

[0069] Diester formation Appropriately, the method is pyrrole A: [ka] or its salts and solvates Pyrrole B: [ka] Or react with its salt and solvate, The following diester compound: [ka] or a step of forming a salt and solvate thereof.

[0070] Appropriately, pyrrole A is, [ka] or its salts and solvates.

[0071] More precisely, R1 is methyl, and the method is pyrrole A: [ka] or its salts and solvates are pyrrole B: [ka] Or react with its salt and solvate, The following diester compound: [ka] The process includes the step of forming a.

[0072] Suitablely, pyrrole A is reacted with pyrrole B in the presence of a protonic acid or montmorillonite clay. Suitablely, pyrrole A is reacted with pyrrole B in the presence of a protonic acid. Suitablely, pyrrole A is reacted with pyrrole B in the presence of montmorillonite clay. Suitablely, the protonic acid is a mineral acid, an organic sulfonic acid, or trifluoroacetic acid. Suitablely, the mineral acid is boric acid, hydrobromic acid, hydrochloric acid, hydrofluoric acid, hydroiodic acid, nitric acid, perchloric acid, phosphoric acid, or sulfuric acid. Suitablely, the organic sulfonic acid is benzenesulfonic acid, camphorsulfonic acid, ethanesulfonic acid, methanesulfonic acid, or p-toluenesulfonic acid. More preferably, the protonic acid is p-toluenesulfonic acid. Suitablely, the montmorillonite clay is K10 montmorillonite clay.

[0073] Ideally, pyrrole A is reacted with pyrrole B in methanol.

[0074] Porphyrin diester formation Appropriately, the method involves the following dicarboxylic acid compound: [ka] Or its salts and solvates are the following dialdehyde compounds: [ka] Alternatively, it can be added to its salts and solvates and reacted to form the following porphyrin diester compound: [ka] The process may also include the step of forming a metallo-derivative, salt, or solvate thereof.

[0075] More precisely, the method involves the following dicarboxylic acid compound: [ka] Or its salts and solvates are the following dialdehyde compounds: [ka] Alternatively, it can be added to its salts and solvates and reacted to form the following porphyrin diester compound: [ka] The process may also include the step of forming a metallo-derivative, salt, or solvate thereof.

[0076] Preferably, (i) a dicarboxylic acid compound or its salt and solvate is added to a dialdehyde compound or its salt and solvate in the presence of a protonic acid or montmorillonite clay. Preferably, this addition is carried out in the presence of a protonic acid. Preferably, this addition is carried out in the presence of montmorillonite clay. Preferably, the protonic acid is a mineral acid, an organic sulfonic acid, or trifluoroacetic acid. Preferably, the mineral acid is boric acid, hydrobromic acid, hydrochloric acid, hydrofluoric acid, hydroiodic acid, nitric acid, perchloric acid, phosphoric acid, or sulfuric acid. Preferably, the organic sulfonic acid is benzenesulfonic acid, camphorsulfonic acid, ethanesulfonic acid, methanesulfonic acid, or p-toluenesulfonic acid. More preferably, the protonic acid is p-toluenesulfonic acid. Preferably, the montmorillonite clay is K10 montmorillonite clay.

[0077] Appropriately, (i) trichloromethane and methanol solvent systems are used to add dicarboxylic acid compounds or their salts and solvates to dialdehyde compounds or their salts and solvates. Appropriately, (i) trichloromethane is the main component (higher than 50% by volume) of the trichloromethane and methanol solvent system. Appropriately, (i) trichloromethane is present in a trichloromethane:methanol ratio of at least 2:1; preferably at least 3:1; preferably at least 4:1. Appropriately, (i) trichloromethane:methanol is present in a ratio of about 5:1.

[0078] Preferably, after adding a dicarboxylic acid compound or its salt and solvate, and a dialdehyde compound or its salt and solvate, (ii) the intermediate is treated with zinc acetate and oxygen; or zinc hexafluoroacetylacetate dihydrate. More preferably, (ii) the intermediate is treated with zinc acetate and oxygen.

[0079] (iii) After treatment with zinc acetate and oxygen, or zinc hexafluoroacetylacetate dihydrate, the intermediate is further treated with trifluoroacetic acid.

[0080] Hydrolysis of porphyrin diesters Appropriately, the method involves the following porphyrin diester compound: [ka] Alternatively, hydrolysis of its metallo derivatives, salts, and solvates yields the dicarboxylic acid of formula (I): [ka] The process further includes the step of obtaining a metallo-dera, salt, or solvate thereof.

[0081] More precisely, the method involves the following porphyrin diester compound: [ka] Alternatively, hydrolysis of its metallo derivatives, salts, and solvates yields the dicarboxylic acid of formula (I): [ka] The process further includes the step of obtaining a metallo-dera, salt, or solvate thereof.

[0082] Ideally, the hydrolysis step is a base hydrolysis step. Ideally, the base hydrolysis step is carried out using sodium hydroxide or lithium hydroxide.

[0083] Ideally, the hydrolysis step is carried out in the presence of tributylammonium chloride (TBAC).

[0084] Ideally, the hydrolysis step is carried out in a dichloromethane and methanol solvent system.

[0085] Ideally, the hydrolysis step is carried out in methanol.

[0086] In some embodiments, hydrolysis is carried out using lithium hydroxide in ethanol.

[0087] Conversion to metallo derivatives Suitablely, the method further comprises the step of converting a compound of formula (I) into a metallo-derivative of the porphyrin dicarboxylic acid of formula (I) or its salts and solvates.

[0088] Suitablely, the metallo-derivative is a tin compound of porphyring dicarboxylic acid of formula (I) or its salts and solvates. More suitablely, the metallo-derivative is a tin mesylate compound, tin oxalate compound, tin phosphate compound, or tin tartrate compound of porphyring dicarboxylic acid of formula (I) or its salts and solvates.

[0089] Preferably, the method further includes the step of converting a compound of formula (I) or its metallo derivative, salt, and solvate to a tin(IV) compound or its salt and solvate. Preferably, the compound of formula (I) or its salt and solvate is brought into contact with a tin source to produce a tin compound of porphyrindicarboxylic acid of formula (I) or its salt and solvate.

[0090] Preferably, the method further comprises the step of converting a compound of formula (I) or its metallo-derivative, salt, and solvate to a metallo-derivative that is a tin compound, the tin compound being a tin acetate compound, tin mesylate compound, tin oxalate compound, tin phosphate compound, or tin tartrate compound of the porphyrindicarboxylic acid of formula (I) or its salts and solvates.

[0091] Preferably, the method further comprises the step of converting a compound of formula (I) or its metallo-derivative, salt, and solvate into a metallo-derivative that is a tin compound, the tin compound being a tin mesylate compound, tin oxalate compound, tin phosphate compound, or tin tartrate compound of the porphyrindicarboxylic acid of formula (I) or its salts and solvates.

[0092] More preferably, the method further includes the step of converting a compound of formula (I) or its metallo derivative, salt, and solvate to a tin(IV) compound of formula (II), (III), (IV), (V), or (VI), or its salt and solvate.

[0093] More preferably, the method further comprises the step of converting a compound of formula (I) or its metallo-derivative, salt, and solvate into a metallo-derivative that is a tin compound, [ka] or its salts and solvates.

[0094] Appropriately, SnO or tin(II) chloride is used to convert the compound of formula (I) or its metallo-derivatives, salts, and solvates into tin(IV) compounds.

[0095] The reaction with SnO is properly carried out in the presence of HCO2H. The reaction with SnO is properly carried out in the presence of CH3CO2H.

[0096] Ideally, the reaction with tin(II) chloride is carried out in the presence of acetic acid. Ideally, the compound of formula (I) or its salt and solvate are refluxed with tin(II) chloride in acetic acid.

[0097] Ideally, the intermediate should be treated with acid after the reaction with SnO. Ideally, the intermediate should be treated with hydrochloric acid after the reaction with SnO.

[0098] In another embodiment, more preferably, the method involves a compound of formula (I) or its metallo-derivative, salt, and solvate, and tin phosphate of formula (II) or (III): [ka] The process further includes the step of converting to a salt or solvate thereof. It is produced by contacting it with a tin source and a phosphate source.

[0099] More preferably, tin(II) pyrophosphate is used to convert the compound of formula (I) or its metallo-derivatives, salts, and solvates into tin(IV) compounds. More preferably, the reaction with tin(II) pyrophosphate is carried out in the presence of acetic acid. More preferably, the compound of formula (I) or its metallo-derivatives, salts, and solvates are refluxed with tin(II) pyrophosphate in acetic acid.

[0100] More appropriately, the method involves using a compound of formula (I) or its metallo-derivative, salt, and solvate as a tin(IV) phosphate salt of formula (VII) or (VIII): [ka] The process further includes the step of converting to a salt or solvate thereof.

[0101] compound More appropriately, the compound of formula (I) or its metallo-derivatives, salts, and solvates are tin salts of the compound of formula (I), and the tin salts are tin acetate, tin mesylate, tin oxalate, tin phosphate, or tin tartrate.

[0102] More appropriately, the tin salt of the compound of formula (I) is stannous mesylate, stannous oxalate, stannous phosphate, or stannous tartrate. More appropriately, the tin salt is stannous mesylate, stannous oxalate, or stannous phosphate, or the tin salt is stannous oxalate, stannous phosphate, or stannous tartrate, or the tin salt is stannous mesylate, stannous phosphate, or stannous tartrate.

[0103] More precisely, the tin salt is tin mesylate or tin phosphate, or the tin salt is tin oxalate, tin phosphate, or tin tartrate.

[0104] The tin salt of the compound of formula (I), wherein the tin salt is tin acetate, tin mesylate, tin oxalate, or tin phosphate.

[0105] More precisely, for any of the above tin salts of the compound of formula (I), substituents R1, R3, R5, and R6 are methyl, and R2 and R4 are ethyl.

[0106] More appropriately, the compounds of formula (I) or their metallo-derivatives, salts, and solvates are the compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates.

[0107] More specifically, the compounds of formula (I) or their metallo-derivatives, salts, and solvates are compounds of formula (II), or (III), or (IV), or (V), or (VI), where substituents R1, R3, R5, and R6 are methyl, and R2 and R4 are ethyl. Such compounds are of formula [ka] It contains JPEG2026509438000043.jpg114170 or its salts and solvates.

[0108] Synthesis of Pyrrole A Appropriately, the method is (i) the compound of the following formula: [ka] Alternatively, by reacting its salt and solvate, an oxime of the following formula can be obtained: [ka] The process further includes the step of producing a salt or solvate thereof.

[0109] More precisely, step (i) is the compound of the following formula: [ka] Alternatively, by reacting its salt and solvate, an oxime of the following formula can be obtained: [ka] Or it produces salts and solvates thereof.

[0110] Ideally, step (i) involves using sodium nitrite and acetic acid to produce an oxime.

[0111] Appropriately, the method involves a cyclization reaction between an oxime or its salt and solvate and R2-CH2C(O)CH3 to produce the pyrrole of the following formula: [ka] The method further comprises (ii) the step of producing a salt and solvate thereof.

[0112] Properly, R2 is acetyl such that R2-CH2C(O)CH3 becomes acetylacetone, and step (ii) of the cyclization reaction of acetylacetone with the oxime or salt and solvate thereof is the pyrrole of the following formula: [ka] Or it produces salts and solvates thereof.

[0113] Ideally, the cyclization reaction in step (ii) is carried out in the presence of one or more of sodium acetate, acetic acid, and zinc.

[0114] Appropriately, the method is (iii) [ka] Alternatively, selective reduction of its salt and solvate yields the following pyrrole compound: [ka] The process further includes the step of producing a salt or solvate thereof.

[0115] Suitably, the selective reduction is a Wolf-Kishner reduction, a Clemmensen reduction, a catalytic hydrogenation, or a reduction using TsNHNH2, in which case it is a NaBH4CN reduction, and such selective reduction is well-known in the following reference 11. Suitably, the Wolf-Kishner reduction is carried out using NH2NH2 and KOH. Suitably, the Clemmensen reduction is carried out using Zn-Hg and HCl. Suitably, the catalytic hydrogenation is carried out using Pt-C under H2 pressure.

[0116] Suitably, the method is (iv) a pyrrole compound of the following formula:

Chemical formula

Chemical formula

[0117] Suitably, step (iv) is catalyzed using lead tetraacetate.

[0118] More suitably, R1 is methyl and step (iv) forms pyrrole A:

Chemical formula

[0119] Synthesis of pyrrole B Suitably, the method is (a) an amine of the following formula:

Chemical formula

Chemical formula

[0120] Appropriately, the method is (b) the isocyanide of the following formula: [ka] Or its salts and solvates, and the alkynes of the following formula: [ka] Alternatively, a cyclization reaction between its salt and solvate may be carried out to produce the following pyrrole compound: [ka] The process further includes the step of forming a salt or solvate thereof.

[0121] More precisely, R4 is C(O)CH3, and the alkyne is given by the following formula: [ka] or it has salts and solvates thereof.

[0122] This type of cyclization reaction is discussed in reference 12 below. Preferably, cyclization step (b) is carried out using a Cu catalyst. Preferably, cyclization step (b) is carried out using a Cu catalyst in the presence of triphenylphosphine (PPh3). Preferably, the Cu catalyst is Cu2O. Preferably, the Cu catalyst is present in amounts of 1-10 mol%; preferably 4-6 mol%; more preferably 5 mol%. Preferably, triphenylphosphine is present in amounts of 10-30 mol%; preferably 15-25 mol%; more preferably 20 mol%. Preferably, R4 is C(O)CH3, and the method is (c) pyrrole compound: [ka] Alternatively, selective reduction of its salt and solvate is performed to obtain pyrrole B: [Chemical formula] Further comprising the step of forming a salt or a solvate thereof.

[0123] More preferably, R3 is methyl, and step (c) is pyrrole B: [[ID=ll]] [Chemical formula] Or form a salt or a solvate thereof.

[0124] Preferably, the selective reduction is Wolf-Kishner reduction, Clemmensen reduction, catalytic hydrogenation, or using TsNHNH2, in which case it is NaBH4CN reduction, and such selective reduction is well-known in the following Reference 11. Preferably, Wolf-Kishner reduction is carried out using NH2NH2 and KOH. Preferably, Clemmensen reduction is carried out using Zn-Hg and HCl. Preferably, catalytic hydrogenation is carried out using Pt-C under H2 pressure.

[0125] Synthesis of dialdehyde Preferably, the method further comprises the step of reacting a compound of the following formula: [Chemical formula] Or a salt or a solvate thereof to react to form an oxime of the following formula: [Chemical formula] Or a salt or a solvate thereof.

[0126] Preferably, step (i) uses sodium nitrite and acetic acid to generate the oxime.

[0127] Preferably, the method uses an oxime or a salt or a solvate thereof and a compound of the following formula: [Chemical formula] The cyclization reaction is carried out, resulting in the pyrrole shown in the following equation: [ka] The method further comprises (ii) the step of producing a salt and solvate thereof.

[0128] Appropriately, the method is to use a compound of the following formula: (ia) [ka] Alternatively, by reacting its salt and solvate, an oxime of the following formula can be obtained: [ka] The process further includes the step of producing a salt or solvate thereof.

[0129] Appropriately, the method involves an oxime or its salt and solvate and a compound of the following formula: [ka] The cyclization reaction is carried out, resulting in the pyrrole shown in the following equation: [ka] The process further includes the step (iia) of generating the result.

[0130] Ideally, the cyclization reaction in step (iia) is carried out in the presence of one or more of sodium acetate, acetic acid, and zinc.

[0131] Appropriately, the method involves the pyrrole compound of the following formula: [ka] Or its salts and solvates, the pyrrole compounds of the following formula: [ka] Alternatively, coupling with its salt and solvate, the following pyrrole diester compounds are formed: [ka] The method further comprises (iii) the step of producing a salt and solvate thereof.

[0132] Appropriately, the method is (i) the compound of the following formula: [ka] Alternatively, by reacting its salt and solvate, an oxime of the following formula can be obtained: [ka] The process further includes the step of producing a salt or solvate thereof.

[0133] Appropriately, the method involves an oxime or its salt and solvate and a compound of the following formula: [ka] The cyclization reaction is carried out, resulting in the pyrrole shown in the following equation: [ka] The method further comprises (ii) the step of producing a salt and solvate thereof.

[0134] Ideally, step (i) or (ia) involves generating an oxime using sodium nitrite and acetic acid.

[0135] Preferably, the cyclization reaction in step (ii) or (iia) is carried out in the presence of one or more of sodium acetate, acetic acid, and zinc. Preferably, the cyclization reaction in step (ii) or (iia) is carried out in the presence of sodium acetate, acetic acid, and zinc.

[0136] Preferably, step (iii) is carried out in methanol, using Br2 as a reagent to produce the pyrrole diester compound. Preferably, step (iii) is carried out in methanol.

[0137] Ideally, step (iii) couples two identical pyrrole compounds (i.e., R5=R6 and R7=R8).

[0138] Appropriately, the method involves the following pyrrole diester compound: [ka] Alternatively, by selective hydrolysis of its salt and solvate, the following pyrrole diacitate compound is obtained: [ka] The method further comprises (iv) the step of producing a salt and solvate thereof.

[0139] Ideally, the selective hydrolysis in step (iv) is carried out under hydrogenation conditions.

[0140] Ideally, the selective hydrolysis in step (iv) is carried out using a Pd / C catalyst.

[0141] The appropriate method is to use the following pyrrole diacid compound: [ka] Or its salts and solvates, the dialdehyde compounds of the following formula: [ka] The method further comprises (v) selective reduction to a salt or solvate thereof.

[0142] R1 R1 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3. Suitablely, R1 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl and C 1-2Selected from alkanamines.

[0143] Appropriately, in some embodiments, R1 is methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3.

[0144] In another embodiment, R1 is H.

[0145] More preferably, R1 is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, and NR'R''. More preferably, R1 is selected from methyl, ethyl, methoxy, and ethoxy.

[0146] More appropriately, R1 is methyl or ethyl. Most appropriately, R1 is methyl.

[0147] R2 R2 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3. Suitablely, R2 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl and C 1-2 Selected from alkanamines.

[0148] Appropriately, in some embodiments, R2 is methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3.

[0149] In another embodiment, R2 is H.

[0150] More preferably, R2 is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', and C(O)CH3. More preferably, R2 is selected from methyl, ethyl, methoxy, ethoxy, and C(O)CH3.

[0151] More appropriately, R2 is methyl or ethyl. Most appropriately, R2 is ethyl.

[0152] R3 R3 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3. Suitablely, R3 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl and C 1-2 Selected from alkanamines.

[0153] Appropriately, in some embodiments, R3 is methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3.

[0154] In another embodiment, R3 is H.

[0155] More preferably, R3 is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, and NR'R''. More preferably, R3 is selected from methyl, ethyl, methoxy, and ethoxy.

[0156] More appropriately, R3 is methyl or ethyl. Most appropriately, R3 is methyl.

[0157] R4 R4 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2Selected from alkanamines and C(O)CH3. Suitablely, R4 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl and C 1-2 Selected from alkanamines.

[0158] Appropriately, in some embodiments, R4 is methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3.

[0159] In another embodiment, R4 is H.

[0160] More preferably, R4 is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', and C(O)CH3. More preferably, R4 is selected from methyl, ethyl, methoxy, ethoxy, and C(O)CH3.

[0161] More appropriately, R4 is methyl or ethyl. Most appropriately, R4 is ethyl.

[0162] R5 R5 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3. Suitablely, R5 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl and C 1-2 Selected from alkanamines.

[0163] Appropriately, in some embodiments, R5 is methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3.

[0164] In another embodiment, R5 is H.

[0165] More preferably, R5 is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, and NR'R''. More preferably, R5 is selected from methyl, ethyl, methoxy, and ethoxy.

[0166] More appropriately, R5 is methyl or ethyl. Most appropriately, R5 is methyl.

[0167] R6 R6 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3. Suitablely, R6 is H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl and C 1-2 Selected from alkanamines.

[0168] Appropriately, in some embodiments, R6 is methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3.

[0169] In another embodiment, R6 is H.

[0170] More preferably, R6 is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, and NR'R''. More preferably, R6 is selected from methyl, ethyl, methoxy, and ethoxy.

[0171] More appropriately, R6 is methyl or ethyl. Most appropriately, R6 is methyl.

[0172] R1, R2, R3, R4, R5, R6 Appropriately, one, two, or three of R1, R2, R3, R4, R5, and R6 are independently H, halogen, methoxy, ethoxy, NR'R'', or C.1-2 Haloalkyl, C 1-2 The compounds are selected from alkanamines and C(O)CH3, and the remainder of R1, R2, R3, R4, R5, and R6 are independently selected from methyl and ethyl.

[0173] Appropriately, one, two, or three of R1, R3, R5, and R6 are independently H, ethyl, halogen, methoxy, ethoxy, NR'R'', or C. 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3, where R1, R3, R5, and the remainder of R6 are methyl, and R2 and R4 are ethyl. More preferably, one of R1, R3, R5, and R6 is independently H, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3, R1, R3, R5, and the remainder of R6 are methyl, and R2 and R4 are ethyl.

[0174] Additionally, as appropriate, optionally, one or two of R1, R3, R5, and R6 may independently be H, ethyl, halogen, methoxy, ethoxy, NR'R'', or C. 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3, where R1, R3, R5, and the remainder of R6 are methyl, and one of R2 and R4 is H, methyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 Selected from alkanamines and C(O)CH3, the other of R2 and R4 is ethyl. Therefore, if no selection is made, R1, R3, R5, and R6 are methyl.

[0175] Additionally, as appropriate, one of R1, R3, R5, and R6 may be independently H, ethyl, halogen, methoxy, ethoxy, NR'R'', or C. 1-2 Haloalkyl, C 1-2Selected from alkanamines and C(O)CH3, R1, R3, R5, and the remainder of R6 are methyl, and R2 and R4 are independently H, methyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl, C 1-2 The group is selected from alkanamines and C(O)CH3. Therefore, if no selection is made, R1, R3, R5, and R6 are methyl.

[0176] In any of the four paragraphs immediately above, the relevant numbers R1, R3, R5, and R6 are independently H, ethyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl and C 1-2 Selected from alkanamines. More preferably, the applicable numbers of R1, R3, R5, and R6 are independently selected from H, ethyl, halogen, methoxy, ethoxy, and NR'R''. More preferably, the applicable numbers of R1, R3, R5, and R6 are independently selected from ethyl, methoxy, and ethoxy.

[0177] In addition, in any of the preceding paragraphs that appropriately begin with, more appropriately, the applicable numbers of R2 and R4 are independently H, methyl, halogen, methoxy, ethoxy, NR'R'', C 1-2 Haloalkyl and C 1-2 Selected from alkanamines. More preferably, the applicable numbers of R2 and R4 are independently selected from H, methyl, halogen, methoxy, ethoxy, and NR'R''. More preferably, the applicable numbers of R2 and R4 are independently selected from methyl, methoxy, and ethoxy.

[0178] More precisely, R1, R3, R5, and R6 are methyl, and R2 and R4 are ethyl.

[0179] R7 R7 is C 1-6 It is alkyl. More precisely, R7 is C 1-5 Selected from alkyl groups. More precisely, R7 is C 1-4Selected from alkyl groups.

[0180] Suitablely, R7 is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, and t-butyl. Suitablely, R7 is selected from methyl, ethyl, n-propyl, and i-propyl. More preferably, R7 is selected from methyl and ethyl. Most preferably, R7 is methyl.

[0181] R8 R8 is C 1-6 It is alkyl. More precisely, R8 is C 1-5 Selected from alkyl groups. More precisely, R8 is C 1-4 Selected from alkyl groups.

[0182] Suitablely, R8 is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, and t-butyl. Suitablely, R8 is selected from methyl, ethyl, n-propyl, and i-propyl. More preferably, R8 is selected from methyl and ethyl. Most preferably, R8 is methyl.

[0183] Strictly speaking, R7 and R8 are the same.

[0184] R9 R9 is selected from benzyl, para-methoxybenzyl, and 2,4-dimethoxybenzyl. More appropriately, R9 is selected from benzyl and para-methoxybenzyl. More appropriately, R9 is benzyl.

[0185] R 10 R 10 The compound is selected from benzyl, para-methoxybenzyl, and 2,4-dimethoxybenzyl. Appropriately, R 10 The compound is selected from benzyl and para-methoxybenzyl. More preferably, R 10 It is benzyl.

[0186] Appropriately, R9 and R10 is the same.

[0187] NR’R’’ In NR’R’’, R’ and R’’ are each independently selected from H and C 1-6 alkyl. Thus, NR’R’’ is NH2, NHC 1-6 alkyl, or N(C 1-6 alkyl)2. Suitably, in some embodiments, NR’R’’ is NH2.

[0188] Suitably, in other embodiments, NR’R’’ is NHC 1-6 alkyl or N(C 1-6 alkyl)2. More suitably, NR’R’’ is NHC 1-6 alkyl. More suitably, NR’R’’ is NHCH3 or NHCH2CH3. More suitably, NR’R’’ is NHCH3.

[0189] Proliferative and / or malignant diseases Compounds of formula (II), or (III), or (IV), or (V), or (VI) or salts and solvates thereof for use in the treatment of proliferative and / or malignant diseases are described.

[0190] Suitably, the proliferative and / or malignant disease can be a metastatic or non-metastatic cancer. The cancer can be a familial or sporadic cancer. Suitably, the proliferative and / or malignant diseases that can be treated can include, for example, benign or intraepithelial lesions and malignant solid tumors, as well as benign and malignant non-solid tumors.

[0191] For example, the proliferative and / or malignant disease can include solid tumors, such as carcinomas or sarcomas. Carcinomas include malignant neoplasms derived from epithelial cells that invade, for example, infiltrate surrounding tissues and give rise to metastases. Adenocarcinomas are carcinomas derived from glandular tissue or tissue that forms recognizable glandular structures.

[0192] The present invention is applicable to the treatment of proliferative and / or malignant diseases.

[0193] In some embodiments, a method is provided for treating a proliferative and / or malignant disease, comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound of the present disclosure or a salt and solvate thereof, or a composition comprising a compound of the present disclosure or a salt and solvate thereof.

[0194] Appropriately, proliferative and / or malignant diseases are selected from breast cancer, lung cancer, brain tumors, and central nervous system cancers, carcinomas, gastrointestinal cancers, hormone-dependent cancers, leukemia, liver cancer, lymphoma, sarcoma and fibrosarcoma, skin cancer and melanoma, urinary tract cancer and genital cancer, and various other cancers.

[0195] Appropriately, proliferative and / or malignant diseases are selected from brain tumors and central nervous system cancers, carcinomas, gastrointestinal cancers, hormone-dependent cancers, leukemias, liver cancers, lymphomas, sarcomas and fibrosarcomas, skin cancers and melanomas, urinary tract cancers and genital cancers, and various other cancers.

[0196] Appropriately treatable brain tumors and central nervous system (CNS) cancers and tumors include astrocytoma (including cerebellar and brain), brainstem glioma, brain tumor, malignant glioma, ependymoma, glioblastoma, medulloblastoma, supratentorial primordial neuroectodermal tumor, visual tract and hypothalamic glioma, primary central nervous system lymphoma, ependymoma, brainstem glioma, visual tract and hypothalamic glioma, extracranial germ cell tumor, medulloblastoma, myelodysplastic syndrome, oligodendroglioma, myelodysplastic / myeloproliferative disorders, myeloid leukemia, myeloid leukemia, multiple myeloma, myeloproliferative disorders, neuroblastoma, plasma cell neoplasm / multiple myeloma, central nervous system lymphoma, true brain tumor, astrocytic brain tumor, glioma, and / or metastatic tumor cell infiltration in the central nervous system.

[0197] The cancers that can be treated include: adrenal cortex, acinar, acinic cells, acinous cells, adenocysts, adenoid cysts, adenoid squamous cells, adenocarcinoma, adenosquamous cell carcinoma, Adnexel, cancer of the adrenal cortex, adrenal cortex, aldosterone production, aldosterone secretion, alveoli, alveolar cells, ameloblasts, ampulla, undifferentiated thyroid carcinoma, apocrine cells, basal cells, basal cells, alveoli, comedone basal cells, cystic basal cells, morpha-like basal cells, multicentric basal cells, nodular ulcerative basal cells, pigmented basal cells, sclerosing basal cells, superficial basal cells. Basaloid, basal squamous epithelial cells, bile duct, extrahepatic bile duct, intrahepatic bile duct, bronchoalveolar, bronchiolar, bronchioloalveolar, bronchoalveolar, bronchoalveolar cells, bronchogenic, rotenocerebral, cholangiocarcinoma, villus, choroid plexus, clear cells, cloaca, anus, colloid, comedo, body, cancer of the body of the uterus, cortisol production, cribriform, castular, castular cells, glandular, glandular, tubular carcinoma of the prostate, ductal carcinoma in situ (DCIS), eccrine, embryonic, armory carcinoma, endometrium, cancer of the endometrium, endometrioid, epidermioid, mixed tumor-derived carcinoma, pleomorphic adenoma-derived carcinoma, lateral growth, fibrous lamellar type, fibrous carcinoma, thyroid follicles Hmm, stomach, gelatinous, gelatinous, giant cell, giant cell carcinoma of the thyroid, giant cell carcinoma, gland, granular cell, hepatocyte, Haasle cell, adrenal gland-like, infant embryonic, islet cell carcinoma, breast cancer, inflammatory breast cancer, carcinoma in situ, intraductal, intraepidermal, intraepithelial, juvenile embryonic, Kruticky cell, large cell, leptomeninges, lobules, invasive lobules, invasive lobules, lobular carcinoma in situ (LCIS), lymphoepithelium, medullary carcinoma, medullary, medullary carcinoma of the thyroid, medullary thyroid, medullary thyroid, melanin, meninges, Merkel cells, degenerate cells, micropapillae, mucinous, musipalm carcinoma, mucinous cell carcinoma, mucinous epithelioid, mucinous carcinoma, mucinous, Nasopharyngeal, cutaneous neuroendocrine carcinoma, non-invasive, non-small cell, non-small cell lung cancer (NSCLC), oat cell, ossifying carcinoma, osteoid, Paget's disease, papillary, papillary thyroid carcinoma, periampuloma, pre-invasive, squamous cell, primary intraosseous, renal cell, scar, schistosomiatic bladder, Schneiderian, hard, sebaceous, signet ring cell, simple carcinoma, small cell, small cell lung cancer (SCLC), spindle cell, cavernous carcinoma, squamous, squamous cell, terminal tubule, undifferentiated thyroid, follicular thyroid, medullary thyroid, papillary thyroid, cutaneous cord-like carcinoma, transitional cell, tubular, undifferentiated thyroid carcinoma, uterine body, wart-like, villous, choriocarcinoma, yolk sac,In particular, this includes squamous cell cancers of the head and neck, squamous cell cancers of the esophagus, and / or oral cancers and carcinomas.

[0198] Treatable gastrointestinal cancers include extrahepatic bile duct cancer, intestinal cancer, colon cancer, colon and rectal cancer, colorectal cancer, gallbladder cancer, gastric (stomach) cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, bladder cancer, islet cell carcinoma (pancreatic endocrine part), pancreatic cancer, islet cell carcinoma, prostate cancer, rectal cancer, salivary gland cancer, small intestine cancer, colon cancer, and polyps associated with colorectal neoplasia. Cancers of the lung and respiratory system that can be treated include bronchial adenoma / carcinoid, esophageal cancer, esophageal cancer, hypopharyngeal cancer, laryngeal cancer, hypopharyngeal cancer, pulmonary carcinoid tumor, non-small cell lung cancer, small cell lung cancer, small cell lung cancer, mesothelioma, nasal and paranasal sinus cancer, nasopharyngeal cancer, oral cancer, oral and lip cancer, oropharyngeal cancer; paranasal and nasal sinus cancer, and / or pleuropneumonoma.

[0199] Hormonal cancers that can be treated include parathyroid cancer, pineal and supratentorial primitive neuroectodermal tumors, pituitary tumors, thymoma and thymic carcinoma, thymoma, thymic carcinoma, thyroid cancer, adrenal cortical cancer, and / or ACTH-producing tumors.

[0200] Leukemias that can be targeted (and may also be considered to include other hematological malignancies) include acute lymphoblastic, acute myeloid, acute lymphoblastic, acute myelogenous leukemia, chronic myeloid, hair follicle, erythroleukemia, lymphoblastic, myeloid, lymphocytic, myelogenous, leukemia, hair follicle, T cell, monocytic, myeloblastic, granulocytic, Gross, mirror-like cell, basophilic, hematoblastic, histiocytic, leukopenic, lymphoid, Schilling, stem cell, myelomonocytic, monocytic, prolymphoblastic, promyelocytic, micromyeloblastic, megakaryoblastic. Megakaryotic, leader cell, bovine, non-leukemic, mast cell, myeloid, plasma cell, subleukemic, multiple myeloma, non-lymphocytic, chronic myeloid leukemia, chronic lymphocytic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin lymphoma (low- and high-grade forms), multiple myeloma, Waldenström macroglobulinemia, heavy chain disease, myelodysplastic syndromes, and myelodysplastic and / or chronic myeloid leukemia.

[0201] Liver cancers that can be targeted include extrahepatic cholangiocarcinoma and / or hepatocellular carcinoma.

[0202] The lymphomas that can be treated include AIDS-related, non-Hodgkin, Hodgkin, T-cell, T-cell leukemia / lymphoma, African, B-cell, B-cell monocyte-like, bovine malignant, Burkitt, central cell, cutaneous lymphoma, diffuse; diffuse large cell; diffuse small cell and large cell mixed type; diffuse, cleavage cell; follicular, follicular central cell, follicular cleavage cell and large cell mixed type, follicular large cell dominant type, follicular cleavage cell dominant type, giant follicular, giant follicular, granulomatous, histiocytic, large cell, immunoblastic, large cleavage cell, large non-cleavage cell, Renato, lymphoblastic, and lymphatic. Examples include pacific, intermediate; lymphocytic, moderately differentiated, plasmacytoid; poorly differentiated lymphocytic, small lymphocytic, well-differentiated lymphocytic, bovine lymphoma; MALT, mantle cells, mantle zone, marginal zone, Mediterranean lymphoma, lymphocytic-histiocytic mixed type, nodular, plasmacytoid, pleomorphic, primary central nervous system, primary exudative, small B cells, small incised cells, small unincised cells, T-cell lymphoma; gyrus T cells, cutaneous T cells, small lymphocytic T cells, undefined lymphoma, U cells, undifferentiated, AIDS-related, central nervous system, cutaneous T cells, exudative (coelomic), thymic lymphoma, and / or cutaneous T-cell lymphoma.

[0203] Appropriate proliferative and / or malignant diseases include tumors, sarcomas and fibrosarcomas, in which tumor cells are embedded in fibrous or homogeneous material such as embryonic connective tissue. Sarcomas that can be targeted include adipose, alveolar soft tissue, ameloblastic, avian, staphyloid, staphyloid sarcoma, roanthoma, chondroblastic, clear cell sarcoma of the tendon sheath, clear cell sarcoma of the kidney, embryonic, endometrial stroma, epithelioid, Ewing's, fascia, fibroblastic, avian, giant cell, granulocytic, vascular endothelial, Hodgkin's, idiopathic multiple pigmented hemorrhagic, B-cell immunoblastic sarcoma, T-cell immunoblastic sarcoma, Examples include Jensen's, Kaposi's, Kupffer's, leukocytic, lymphoid, melanistic, mixed cell, multiple, lymphatic, idiopathic, pluripotent primary sarcoma of bone, osteoblastic, osteogenic, paraostemic, pleomorphic, pseudokaposi's, reticular, reticular cell sarcoma of the brain, rhabdomyosarcoma, Routh's, soft tissue, spindle cell, synovial, telangiectatic, bone sarcoma (osteosarcoma) / malignant fibrous histiocytoma, and / or soft tissue sarcoma.

[0204] Treatable skin cancers (including non-melanomas) and melanomas include cutaneous T-cell lymphoma, intraocular melanoma, metastatic malignant melanoma, tumor progression of human cutaneous keratinocytes, basal cell carcinoma, and squamous cell carcinoma. Targetable ocular cancers include intraocular melanoma, retinoblastoma, and / or intraocular melanoma.

[0205] Cancers of the urinary tract and reproductive organs that can be treated include cervical cancer, endometrial cancer, ovarian epithelial cancer, extragonadal germ cell tumors, extracranial germ cell tumors, extragonadal germ cell tumors, ovarian germ cell tumors, gestational trophoblastic neoplasms, spleen cancer, kidney cancer, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumors, low-grade ovarian tumors, penile cancer, kidney cancer, retinocellular carcinoma (including carcinoma), renal cell carcinoma, renal pelvis and ureter (transitional cell carcinoma), transitional cell carcinoma of the renal pelvis and ureter, gestational trophoblastic neoplasms, testicular cancer, ureter and renal pelvis, transitional cell carcinoma, urethral cancer, endometrial uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, ovarian cancer, primary peritoneal epithelial neoplasms, cervical cancer, uterine cancer and solid tumors in the ovarian follicles, superficial bladder tumors, invasive transitional cell carcinoma of the bladder, and / or muscle-invasive bladder cancer.

[0206] Other cancers that can be targeted include advanced cancer, AIDS-related cancer, anal cancer, adrenal cortical cancer, aplastic anemia, aniline cancer, betel nut cancer, bone cancer, gnathomycosis, carcinoid cancer (gastrointestinal and bronchial), Castleman disease, chronic myeloproliferative disorders, rotorous cancer, chimney sweep cancer, clay tube cancer, colloidal cancer, contact cancer, cystic cancer, dendritic cancer, gametocarcinoma, tube cancer, dye worker cancer, cerebral cancer, armory cancer, endometrium, endothelial cancer, epithelial cancer, Ewing tumor family, glandular cancer, head and neck cancer, hemangiovascular cell tumor, carcinoma in situ, kangu cancer, and latent cancer. Examples include labial, lip and oral cancer, medullary, melanistic, melanistic, metastatic cervical squamous cell carcinoma of unknown primary origin, multiple endocrine neoplasm syndrome, multiple myeloma / plasma cell neoplasm, mule spinner, mycosis fungoides, occult cancer, paraffin, peritoneal exudative, malignant pleural effusion, pheochromocytoma, pitch worker, scar, schistosomiasis bladder, scirrhous, Sézary syndrome, lymph node, soft tissue sarcoma, soot, spindle cell, supratentorial primitive neuroectodermal tumor, swamp, tar, tubular carcinoma, chorionic neoplasm, trophoblastic neoplasm, and Wilms' tumor.

[0207] More appropriately, proliferative and / or malignant diseases include bladder cancer, brain tumors, breast cancer, hepatocellular carcinoma, acute myeloid leukemia, lung cancer, non-small cell lung cancer, non-Hodgkin lymphoma, glioma, glioblastoma, melanoma, metastatic melanoma, ovarian cancer, or prostate cancer.

[0208] Appropriately, the compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, are administered alone or concurrently or sequentially with one or more additional therapeutic agents.

[0209] Appropriately, one or more additional therapeutic agents are selected from one or more immunotherapies or other anticancer drugs.

[0210] Appropriately, immunotherapeutic agents may include one or more cancer vaccines and / or therapeutic antibodies and / or small molecules, and / or RNA / DNA-based agents, and / or viral agents (such as oncolytic viruses or carriers), and / or cell therapies or genetically modified cell therapies. Cancer vaccines are agents, cell-based agents, molecules, or immunogens that stimulate, utilize, or induce an endogenous immune response to one or more tumor antigens in an individual or subject.

[0211] Cell therapy or genetically modified cell therapy includes adoptive cell therapies such as TIL therapy, CAR-T therapy, and other immune effector cell therapies. In some embodiments, immune effector cells include or consist of NK cells, T cells, B cells, dendritic cells, macrophages, peripheral blood mononuclear cells (PBMCs), ab T cells, gd T cells, regulatory T cells, NK T cells, or mesenchymal cells, or combinations thereof.

[0212] In this specification, tumor antigens are broadly defined as antigens expressed within or specifically by tumor or cancer cells. Antigens may be expressed on the cell surface that are recognized by components of the humoral immune system, such as B lymphocytes (B cells). Intracellular tumor antigens (including those associated with secreted cytoplasmic and nuclear proteins) may be processed into shorter peptide fragments to form complexes with major histocompatibility complex (MHC) molecules I and II, also known as human leukocyte antigens (HLA) (presented on the cell surface of cancer cells and recognized by T cell receptors (TCFs) of T lymphocytes (T cells)). Antigens may also include exogenous and autologous lipids (such as those presented on CD1 molecules) or abnormal glycosylation or post-translational modifications. Preferably, tumor antigens are not expressed by normal cells, or at least not at the same level as those expressed within tumor cells. Cancer vaccines currently produced and under development for human treatment activate either the humoral immune system (i.e., the antibody-dependent immune response) or the cell-mediated immune system, which includes T lymphocytes (T cells) that can specifically recognize and kill tumor cells.

[0213] Cancer vaccines can enhance the presentation of one or more tumor antigens to both antigen-presenting cells (e.g., macrophages and dendritic cells) and / or other immune cells such as T cells, B cells, and NK cells. In some cases, preparations and / or formulations of cancer vaccines can be used with one or more adjuvants known in the art to induce or enhance the immune response.

[0214] An adjuvant is a substance incorporated into or administered with an antigen to enhance the immune response. Adjuvants can improve the immunological response by providing an antigen reservoir (extracellular or intracellular in macrophages / DCs) and activating antigen-presenting cells to stimulate specific lymphocyte populations. Many types of adjuvants are well known in the art. Specific examples of adjuvants include monophosphoryl lipid A (MPL, SmithKline Beecham), which is a similar product obtained after purification and acid hydrolysis of Salmonella Minnesota Re 595 lipopolysaccharide; saponins, including QS21 (SmithKline Beecham), a high-purity QA-21 saponin purified from Quillaja saponaria extract; DQS21 (SmithKline Beecham), described in International Publication No. 96 / 33739; QS-7, QS-17, QS-18, and QS-L1 (So et al., MoI Cells (1997) 7: 178-186); and ISCOMATRIX adjuvants, which are cage-like structures composed of saponins, phospholipids, and cholesterol (e.g., Maraskovsky et al., Clin. Cancer Res. (2004)). Examples include 10:2879-2890; incomplete Freund adjuvants; complete Freund adjuvants; montanides; alum; CpG oligonucleotides (see, e.g., Kreig et al., Nature 374:546-9, 1995) and other immunostimulant oligonucleotides, including poly-IC and poly-ICLC (Hiltonol®); and various water-in-oil emulsions prepared from biodegradable oils such as squalene and / or tocopherol.

[0215] Appropriately, cancer cells in an individual express tumor antigens that immunologically cross-react with cancer vaccines. Appropriately, tumor antigens are expressed in cancer cells in an individual but not in normal somatic cells. Appropriate tumor antigens for use in immunotherapies such as cancer vaccines include P1A, MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A5, MAGE-A6, MAGE-A7, MAGE-A8, MAGE-A9, MAGE-A10, MAGE-A11, MAGE-A12, GAGE-I, GAGE-2, GAGE-3, GAGE-4, GAGE-5, GAGE-6, GAGE-7, GAGE-8, BAGE-I, RAGE-1, L Examples include B33 / MUM-1, FRAME, NAG, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), tyrosinase, cerebral glycogen phosphorylase, Melan-A, MAGE-C1 / CT7, MAGE-C2, NY-ESO-I, LAGE-I, SSX-I, SSX-2 (HOM-MEL-40), SSX-3, SSX-4, SSX-5, SCP-I, and XAGE, as well as their immunogenic fragments.Other types of tumor antigens include overexpressed or mutant proteins and differentiation antigens, particularly melanocyte differentiation antigens, such as p53, ras, CEA, MUC1, PMSA, PSA, tyrosinase, melan-A, MART-1, gp100, gp75, α-actinin-4, Bcr-Abl fusion protein, Casp-8, β-catenin, cdc27, cdk4, cdkn2a, coa-1, dek-can fusion protein, EF2, ETV6-AML1 fusion protein, LDLR-fucosyltransferase AS fusion protein, HLA-A2, HLA-A11, hsp70-2, KIAAO205, Mart2, Mum- 2 and 3, neo-PAP, myosin class I, OS-9, pml-RARα, fusion protein, PTPRK, K-ras, N-ras, triose phosphate isomerase, GnTV, Herv-K-mel, NA-88, SP17, TRP2-Int2, (MART-I), E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein-Barr virus antigen, EBNA, human papillomavirus (HPV) antigens E6 and E7, TSP-180, MAGE-4, MAGE-5, MAGE-6, p185erbB2, p180erbB-3, c-met, nm-23H1, PSA, TAG-72-4, CA Examples include 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, α-fetoprotein, 13HCG, BCA225, BTAA, CA 125, CA 15-3 (CA 27.29\BCAA), CA 195, CA 242, CA-50, CAM43, CD68\KP1, CO-029, FGF-5, G250, Ga733 (EpCAM), HTgp-175, M344, MA-50, MG7-Ag, MOV18, NB\170K, NY-CO-1, RCAS1, SDCCAG16, TA-90 (Mac-2 binding protein\cyclophyllin C-related protein), TAAL6, TAG72, TLP, and TPS, as well as tyrosinase-related proteins such as TRP-1 and TRP-2.Other suitable antigens include the following classes of cancer antigens outlined in Cancer Vaccines and Immunotherapy (2000) Eds Stern, Beverley and Carroll, Cambridge University Press, Cambridge: cancer-testis antigens (e.g., HOM-MEL-40), differentiation antigens (e.g., HOM-MEL-55), overexpression gene products (HOM-MD-21), mutant gene products (NY-COL-2), splice variants (HOM-MD-397), splice peptides (13, 14, 15), gene amplification products (HOM-NSCLC-11), and cancer-associated autoantigens (HOM-MEL-2.4). In some exemplary embodiments, the antigen is a tumor antigen selected from the group consisting of MUC1, MAGE, BAGE, RAGE, CAGE, SSX-2, NY-ESO-1, FRAME, PSMA, tyrosinase, Melane-A, and mixtures thereof. In some variant forms, the cancer antigen is a mammalian protein. In some variants, cancer antigens are human proteins. In some variants, full-length proteins can be used as antigens. In some variants, peptides containing antigenic fragments of these proteins can be used as tumor antigens.

[0216] Other suitable tumor antigens are well known in the art (see, for example, International Publication No. 00 / 20581). The sequences of these tumor antigens are readily available from public databases, but can also be found in International Publications No. 1992 / 020356, No. 1994 / 005304, No. 1994 / 023031, No. 1995 / 020974, No. 1995 / 023874, and No. 1996 / 026214.

[0217] In some embodiments, a method of treating a subject using compounds of formula (II), (III), (IV), (V), or (VI) or their salts and solvates, and immunotherapeutic agents disclosed herein, may further include a step of treating the tumor by administering one or more tumor treatments. Such treatments include, for example, oncological drugs, radiation, and surgical procedures.

[0218] In this specification, “oncology drugs” refers to drugs administered to a target for the purpose of treating cancer. Various types of drugs for treating tumors are described herein.

[0219] Oncology drugs function in various ways. Some cancer drugs work by targeting physiological mechanisms specific to tumor cells. Examples include targeting specific genes and their gene products (i.e., essentially proteins) that are mutated in tumors. Such genes include, but are not limited to, oncogenes (e.g., Ras, Her2, bcl-2), tumor suppressor genes (e.g., EGF, p53, Rb), and cell cycle targets (e.g., CDK4, p21, telomerase). Oncology drugs can alternately target signaling pathways and other molecular mechanisms that are altered in tumor cells.

[0220] Examples of immunotherapeutic agents for use in combination with the compounds described herein include biological response modifiers. Examples of biological response modifiers for use in the combination therapies disclosed herein include interferon-α, IL-2, G-CSF, and GM-CSF; various agents, such as platinum-coordinated complexes including cisplatin and carboplatin; anthracendions such as mitoxantrone; substituted ureas such as hydroxyurea; methylhydrazine derivatives including N-methylhydrazine (MIH) and procarbazine; corticosteroids such as mitotane (o,p'-DDD) and aminoglutethimide; antagonists, including hormones and corticosteroid antagonists, such as prednisone and its equivalents, dexamethasone, and aminoglutethimide; progestins such as hydroxyprogesterone caproate, medroxyprogesterone acetate, and megestrol acetate; and diethyl Examples include estrogens such as stilbestrol and ethinylestradiol equivalents; anti-estrogens such as tamoxifen; androgens including testosterone propionate and fluoxymesterone / equivalents; anti-androgens such as flutamide, gonadotropin-releasing hormone analogs, and leuprolide; nonsteroidal antiandrogens such as flutamide; kinase inhibitors, histone deacetylase inhibitors, methylation inhibitors, proteasome inhibitors, oxidants, antioxidants, telomerase inhibitors, BH3 mimetic agents, ubiquitin ligase inhibitors, and STAT inhibitors; receptor tyrosine kinase inhibitors such as imatinib (Gleevec or Glivec); and erlotinib (EGF receptor inhibitor), currently marketed as Tarceva.

[0221] Additional biological response modifiers include inhibitors of the immunomodulatory enzymes indoleamine 2,3-dioxygenase (IDO or INDO EC 1.13.11.52) and tryptophan 2,3-dioxygenase (TDO, EC 1.13.11.11).

[0222] Tryptophan 2,3-dioxygenase (TDO) is a homotetrameric heme-containing cytosolic enzyme encoded by the gene TD02 and highly expressed in the liver. It catalyzes the first rate-limiting step in tryptophan degradation along the kynurenine pathway, thereby regulating systemic tryptophan levels. The role of TDO in cancer was demonstrated in a study by Pilotte et al. (Proc Natl Acad Sci US A. 2012 Feb 14; 109(7): 2497-502. Epub 2012 Jan 30). These studies detected TDO expression in a significant proportion of human tumors. In preclinical models, TDO expression by tumors prevented their rejection by immune mice. Studies have shown that TDO inhibitors, when used systemically, restored the mice's ability to reject TDO-expressing tumors, thus providing evidence that TDO inhibitors may be effective in cancer treatment. LM10 is one example of a TDO inhibitor.

[0223] Indoleamine 2,3-dioxygenase (IDO) is an enzyme encoded by the ID01 gene in humans. This enzyme also catalyzes the first rate-limiting step in the breakdown of the essential amino acid L-tryptophan to N-formylkynurenine and is normally expressed in tumor cells and activated immune cells. IDO weakens the immune response by breaking down the indole moiety of tryptophan, drastically reducing tryptophan levels locally and increasing pro-apoptotic kynurenine. As a result, IDO blocks the proliferation and activation of T cells that are extremely sensitive to Trp deficiency. This creates an environment in which tumor-specific cytotoxic T lymphocytes become functionally inactive or can no longer attack the patient's cancer cells. The finding that many human tumors constitutively express IDO has led to the hypothesis that inhibiting it could improve the effectiveness of cancer immunotherapy. Results from in vitro and in vivo studies show that the efficacy of therapeutic vaccination in cancer patients can indeed be improved by the co-administration of IDO inhibitors.

[0224] Small molecule inhibitors of IDOs are available in the art to treat IDO-related diseases such as cancer. For example, International Publication 99 / 29310 reports a method for modifying T cell-mediated immunity, comprising the step of modifying, in particular, the local extracellular concentrations of tryptophan and tryptophan metabolites. Additional compounds having IDO inhibitory activity are reported in International Publication 2004 / 094409, U.S. Patent No. 8088803 (which reports compound INCB024360), U.S. Patent Application Publication 20110165188, and U.S. Patent Application Publication 20110159017. Cytokines can be used as immunotherapeutic agents for use in combination with the compounds described herein.

[0225] Cytokines effective in inhibiting tumor growth / metastasis can be used in combination with the compounds described herein. Such cytokines, lymphokines, or other hematopoietic factors include, but are not limited to, M-CSF, GM-CSF, TNF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IFN (types I and II), TNFα, TNF1, TNF2, G-CSF, Meg-CSF, GM-CSF, thrombopoietin, stem cell factors, and erythropoietin.

[0226] In this specification, the terms “immunotherapy,” “immunotherapy agent,” or “immunotherapy drug” generally refer to any therapeutic method aimed at treating or curing a disease, particularly cancer, by mobilizing or manipulating a patient’s immune system. The terms “immunotherapy,” “immunotherapy agent,” or “immunotherapy drug” encompass the targeting of tumor cells via the recognition of immunogenic proteins or antigens expressed by tumor cells, achieved by utilizing passively transmitted immune molecules such as antibodies, or cancer vaccine preparations designed to induce antibodies or T lymphocytes (T cells) that recognize local areas of antigens or epitopes specific to tumor cells.

[0227] The immunotherapeutic agent intended for use in combination with the compounds described herein may be an antibody.

[0228] Appropriately, therapeutic antibodies include one or more anti-Her2 / Neu receptor antibodies, such as trastuzumab (marketed as Herceptin); alemtuzumab, a CD52 antibody currently under development as Lemtrada, marketed as Campath, MabCampath, or Campath-1H; gemtuzumab, an anti-CD33 monoclonal antibody linked to calicheamicin, marketed as Mylotarg by Wyeth; anti-CD20 antibodies such as rituximab (marketed as Rituxan and MabThera) or ibritumomab tiuxetan marketed under the trade name Zevalin; infliximab (marketed as Remicade), or adalimumab (marketed as Humira), or ethanol. This includes anti-TNF-α antibodies such as soluble TNFR2 molecules like Nercept (also known as Enbrel); antibodies against the CD25 chain of the IL-2 receptor, such as basiliximab (trade name Simulect); anti-CD40 / CD40L antibodies such as humanized IgGl anti-human CD40 antibody (SGN-40); anti-CTLA-4 blocking antibodies such as ipilimumab (also known as MDX-101 or MDX-010, marketed as Yervoy) or tremelimumab; anti-PD-1 antibodies (programmed cell death protein 1, also named CD279); anti-PDL-1 (programmed cell death ligand); antibodies against glucocorticoid-inducible TNFR family-related genes, or anti-GITR antibodies; or anti-OX-40 (CD134) antibodies.

[0229] Other suitable immunotherapeutic agents include soluble lymphocyte-activating gene 3 (also known as LAG3 or CD223) immunomodulators such as LAG3-Ig (IMP321); Toll-like receptor agonists such as MPL, CpG, single-stranded RNA, nucleotides, nucleotide analogs, CL087 (TLR7-specific ligand), loxolibine, polyinosine-polycytidylic acid, flagellin, reximod, immiquimod, and gardiquimod; NOD ligands such as muramyl dipeptide, murabutide, peptidoglycan, and muramyl dipeptide; and antiviral agents such as oseltamivir phosphate, amphotericin B, and palivizumab.

[0230] In some embodiments, the therapies disclosed herein may include the administration of compounds described herein, such as compounds of formula (II), (III), (IV), (V), or (VI), or salts and solvates thereof, and one or more antibodies selected from the group consisting of anti-PD1 antibodies, anti-PDL-1 antibodies, anti-CTLA-4 antibodies, anti-GITR antibodies, and anti-OX40 antibodies.

[0231] Anti-PD1 antibodies can be monoclonal antibodies produced against the negative immunomodulatory human cell surface receptor PD-1, which possesses immunoenhancing activity. An exemplary anti-PD1 antibody is the human monoclonal antibody MDX-1106, which binds and blocks the activation of PD-1 by its ligands PD-L1 and PD-L2, resulting in T cell activation and a cell-mediated immune response against tumor cells. In some embodiments, anti-PD-1 antibodies are monoclonal antibodies produced against the protein ligand PD-L1, which possesses immunomodulatory and antitumor activity. An exemplary anti-PD-L1 antibody is the human monoclonal antibody MDX-1105, which binds PD-L1 and blocks its binding to and activation of its receptor PD-1, thereby improving the T cell-mediated immune response against neoplasms and restoring T cell inactivation in chronic infections. PD-L1 is widely expressed in hematopoiesis and parenchymal tissues.

[0232] Anti-CTLA-4 antibodies can be monoclonal antibodies formulated against the T cell receptor protein cytotoxic T lymphocyte-associated protein 4 (CTLA-4). An exemplary anti-CTLA-4 antibody is the human IgG2 monoclonal antibody tremelimumab, which binds to CTLA4 and blocks the binding of antigen-presenting cell ligands B7-1 and B7-2 to CTLA-4, thereby inhibiting the B7-CTLA4-mediated downregulation of T cell activation. Another exemplary anti-CTLA-4 antibody is the human IgG1 monoclonal antibody ipilimumab, which binds to CTLA4 and blocks the binding of antigen-presenting cell ligands B7-1 and B7-2 to CTLA-4, thereby inhibiting the B7-CTLA4-mediated downregulation of T cell activation. Ipilimumab is in clinical trials for the treatment of non-small cell lung cancer, small cell lung cancer, and metastatic hormone-refractory prostate cancer.

[0233] Anti-GITR antibodies are monoclonal antibodies produced against glucocorticoid-induced tumor necrosis factor receptor (GITR), which block the interaction between GITR and its ligand, enhance the cytotoxicity of native human killer cells, and / or downregulate GITR expression on peripheral blood lymphocytes.

[0234] Anti-OX40 antibodies can be agonist monoclonal antibodies that mimic the natural OX40 ligand, selectively bind to the OX40 receptor, and activate it. Receptor activation induces the proliferation of memory and effector T cells.

[0235] Appropriately, anticancer agents may include any known agents having desirable anticancer properties. Appropriately, anticancer agents may include one or more of the following: taxoids such as Taxol® and Taxotere®, Abraxane, or other chemotherapeutic agents such as cisplatin (and other platin inserts), etoposide and etoposide phosphate, bleomycin, mitomycin C, CCNU, doxorubicin, daunorubicin, idarubicin, and ifosfamide.Other anticancer agents may be considered for use in the combination therapies disclosed herein, including aspirin, sulindac, curcumin; mechloretamine, cyclophosphamide, ifosfamide, melphalan, and nitrogen mustards such as chlorambucil; nitrosoureas such as carmustine (BCNU), lomustine (CCNU), and semustine (methyl-CCNU); triethylenemelamine (TEM), Alkylating agents including ethyleneimine / methylmelamines such as ethylene, thiophosphoramide (thiotepa), hexamethylmelamine (HMM, altoretamine), alkyl sulfonates such as busulfan, and triazines such as dacarbazine (DTIC); folic acid analogs such as methotrexate and trimethrexate, 5-fluorouracil, fluorodeoxyuridine, gemcitabine, cytosine arabinoside (AraC, cytarabine), 5-azacitidine, 2,2'-diphthol Antimetabolites including pyrimidine analogs such as ruorodeoxycytidine, purine analogs such as 6-mercaptopurine, 6-thioguanine, azathiopurine, 2'-deoxycoformycin (pentostatin), erythrohydroxynonyladenine (EHNA), fludarabine phosphate, and 2-chlorodeoxyadenosine (cladribine, 2-CdA); natural products including antimitotics such as paclitaxel, vinblastine (VLB), vincristine, and vinorelbine. Examples include vinca alkaloids, taxotere, estramustine, and estramustine phosphate; epipodophyllotoxins such as etoposide and teniposide; antibiotics such as actinomycin D, daunomycin (rubidomycin), doxorubicin, mitoxantrone, idarubicin, bleomycin, plicamycin (mitramycin), mitomycin C, and actinomycin; and enzymes such as L-asparaginase.Anticancer drugs can also be biological agents such as proteins that inhibit tumor growth, including interferon (IFN)-γ, tumor necrosis factor (TNF)-α, TNF-β, GM-CSF, and similar cytokines; or anti-angiogenic factors such as angiostatins and endostatins; or soluble forms of angiogenic factor receptors, including soluble VGF / VEGF receptors, or inhibitors of FGF or VEGF. Further anticancer drugs include platinum-coordinated complexes such as cisplatin and carboplatin, anthracendions such as mitoxantrone, substituted ureas such as hydroxyurea, methylhydrazine derivatives including N-methylhydrazine (MIH) and procarbazine, corticosteroids such as mitotane (o,p'-DDD) and aminoglutethimide; hormones and antagonists including prednisone and equivalents, dexamethasone and aminoglutethimide; progestins such as hydroxyprogesterone caproate, medroxyprogesterone acetate and megestrol acetate; estrogens such as diethylstilbestrol and ethinylestradiol equivalents; antiestrogens such as tamoxifen; and propionate tessyl estrone. Androgens, including tosterone and fluoxymesterone / equivalence; antiandrogenic drugs such as flutamide, gonadotropin-releasing hormone analogs and leuprolide; nonsteroidal antiandrogens such as flutamide; kinase inhibitors, histone deacetylase inhibitors, methylation inhibitors, proteasome inhibitors, monoclonal antibodies, oxidants, antioxidants, telomerase inhibitors, BH3 mimetic agents, ubiquitin ligase inhibitors, STAT inhibitors, and receptor tyrosine kinase inhibitors such as imatinib mesylate (marketed as Gleevac or Glivac) and erlotinib (EGF receptor inhibitor), currently marketed as Tarveca; and antiviral drugs such as oseltamivir phosphate, amphotericin B, and palivizumab. STING agonists such as DMXAA are also included.

[0236] More appropriately, the anticancer agent comprises one or more pyrimidine analogs and antimitotic agents. More appropriately, the anticancer agent comprises one or more of the following: 5-fluorouracil, fluorodeoxyuridine, gemcitabine, cytosine arabinoside (AraC, cytarabine), 5-azacitidine, 2,2'-difluorodeoxycytidine, paclitaxel, vinblastine (VLB), vincristine, vinorelbine, taxotere, estramustine, and estramustine phosphate.

[0237] Appropriately, the anticancer agent may further contain an antibody that directly or indirectly conjugates to one or more agents having desirable anticancer properties to form an antibody-drug conjugate. A suitable antibody can be selected to direct the conjugate to desired cells, such as tumor cells. Such antibody-drug conjugates are well known in the art (see reference 16 below).

[0238] This treatment can treat any type of cell, including but not limited to cells of the bones, eyes, head and neck, lungs, gastrointestinal tract (e.g., mouth, esophagus, intestines, colon), chest (breasts), neck, ovaries, uterus, prostate, liver, kidneys, bladder, pancreas, brain, and skin.

[0239] Those skilled in the art can easily determine whether a candidate compound treats the proliferative and / or malignant condition of any particular type of cell.

[0240] More appropriately, the subjects are humans, domesticated animals, and companion animals. More appropriately, the subjects are humans.

[0241] Metastasis inhibitors Compounds of formula (I) or their metallo-derivatives, salts, and solvates, wherein the compound is a compound of formula (II), (III), (IV), (V), or (VI), or its salts and solvates, for example, compounds of formula (VII), (VIII), (IX), (X), or (XI), or their salts and solvates, have been shown to have transfer inhibitory activity.

[0242] Therefore, compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, may be used in the manufacture of pharmaceuticals for treating or inhibiting metastasis.

[0243] More appropriately, the compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, for use in treating or inhibiting metastasis, are the compounds of formula (VII), (VIII), (IX), (X), or (XI), or their salts and solvates. More appropriately, the compounds are the compounds of formula (II) or (III), or their salts and solvates. More appropriately, the compounds are the compounds of formula (VII) or (VIII), or their salts and solvates.

[0244] Treatment of metastasis includes the step of administering a therapeutically effective amount of a compound or pharmaceutical composition described herein to a subject in need.

[0245] Appropriately, metastases originate from bladder cancer, breast cancer, colorectal cancer, esophageal cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, or stomach cancer, or any combination thereof.

[0246] The spread of cancer cells from the primary tumor site to distal organs is called metastasis. Cancer tumor metastasis is a major cause of treatment failure when treating a disease, as patients succumb to the proliferation of multiple tumors. The extent to which metastasis occurs varies depending on the individual tumor type. Melanoma, breast cancer, lung cancer, and prostate cancer are particularly prone to metastasis.

[0247] When metastasis occurs, secondary tumors can form in various parts of the body, with the lungs being one of the more common sites of metastasis.

[0248] Therefore, any degree of inhibition of tumor metastasis is beneficial, regardless of the effect the inhibitory agent has on the primary tumor. However, if the agent also inhibits the primary tumor, this is an additional benefit of the agent.

[0249] The metastasis inhibitors disclosed herein may be used alone or in combination as part of a treatment regimen for animal or human patients with metastatic cancers, particularly melanoma, breast cancer, lung cancer, and prostate cancer. Treatments that inhibit metastasis formation are best administered as soon as possible after cancer detection. By utilizing treatment regimens in early-stage patients, the treating physician maximizes the chances that significant metastasis has not yet occurred. This maximizes the chances of a successful treatment. In such regimens, metastasis inhibitors or salts thereof may, and are generally, administered in combination with other forms of treatment that control the primary tumor itself.

[0250] Appropriately, compounds of formula (I) or their metallo-derivatives, salts, and solvates for use in treating or inhibiting metastasis are administered concurrently or sequentially with one or more immunotherapeutic agents or other anticancer agents.

[0251] The treatments described herein may also be used in conjunction with (i.e., immediately before or after) a surgical procedure to remove primary tumor material from the body. Often, surgical procedures to remove tumor material from the body are avoided due to the fear that metastasis of tumor cells may occur as a result of such physical procedures. However, if the metastasis inhibitors described herein are administered to the patient prior to the surgical procedure, the risk of metastasis that may occur as a result of surgery can be reduced, making surgery a more attractive treatment option.

[0252] Within reasonable limits of medical judgment, the dosage and method of administration of the metastasis inhibitors described herein will vary depending on the severity and nature of the specific condition being treated, the duration of treatment, the adjunctive therapies used, the patient's age and health status, and similar factors within the specific knowledge and expertise of the attending physician. However, single doses can typically range from 0.01 to 2000 milligrams per kilogram of body weight, preferably 1 to 200 milligrams per kilogram (unless otherwise specified, units designated as "mg / kg" herein refer to milligrams per kilogram of body weight). Up to four doses per day can be used routinely, but this can be adjusted according to the patient's needs while maintaining a reasonable benefit / risk ratio. Variations in patient response are to be expected, but the higher dose within the indicated range is usually required for oral administration, and the lower dose applies to intravenous administration.

[0253] Pharmaceutical composition Appropriately, the pharmaceutical composition further comprises one or more additional therapeutic agents.

[0254] Appropriately, the pharmaceutical composition further comprises one or more immunotherapeutic agents or one or more additional therapeutic agents selected from one or more immunotherapeutic agents or other anticancer agents.

[0255] Appropriately, the pharmaceutical composition further comprises one or more other anticancer agents.

[0256] Preferably, the pharmaceutical composition further comprises one or more cancer vaccines or therapeutic antibodies. Preferably, the pharmaceutical composition further comprises an adjuvant.

[0257] Appropriately, the pharmaceutical composition comprises a compound of formula (II), (III), (IV), (V), or (VI) or a salt or solvate thereof, a cancer vaccine or therapeutic antibody, an adjuvant, and pharmaceutically acceptable excipients, carriers, or diluents.

[0258] Ideally, the pharmaceutical composition is formulated as a nanoparticle carrier medium or a viral vector.

[0259] Suitablely, the pharmaceutical composition is formulated as a nanoparticle carrier medium. Suitablely, the nanoparticle carrier medium is one of silica nanoparticles, liposomes, micelles, nanogels, or polymer nanoparticles. Suitablely, the nanoparticle carrier medium is liposomes.

[0260] Appropriately, the pharmaceutical composition is formulated as a viral vector. Appropriately, the viral vector is an adenovirus, adeno-associated virus, retrovirus, lentivirus, or herpesvirus vector. The use of viral vectors is a well-known method of delivery (see references 17 and 18 below).

[0261] Dosage and Administration Compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, may be administered alone or in combination with any one or more compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, and other pharmacologically active compounds.

[0262] The compounds of the present invention can be appropriately used in combination with various components to produce compositions of the present invention. The compositions can be appropriately used in combination with pharmaceutically acceptable carriers or diluents to produce pharmaceutical compositions (which may be for human or animal use). Suitable carriers and diluents include isotonic saline solutions (e.g., phosphate-buffered saline), water, ethanol, propylene glycol, glycerin, and combinations thereof. Useful pharmaceutical compositions and methods for their preparation can be found in standard pharmaceutical textbooks. For example, see Handbook for Pharmaceutical Additives, 3rd Edition (eds. M. Ash and I. Ash), 2007 (Synapse Information Resources, Inc., Endicott, New York, USA) and Remington: The Science and Practice of Pharmacy, 21st Edition (ed. DB Troy) 2006 (Lippincott, Williams and Wilkins, Philadelphia, USA), which are incorporated herein by reference.

[0263] The compounds of the present invention can be administered by any suitable route. Preferably, the compounds of the present invention are administered orally or by any parenteral route in the form of a pharmaceutically acceptable dosage form, in the form of a pharmaceutical preparation containing the active ingredient in any form of a non-toxic organic or inorganic acid or base, or an addition salt. Preferably, the parenteral route is selected from intramuscular, subcutaneous, intravenous, and intradermal administration. More preferably, the parenteral route is selected from intramuscular and intravenous administration.

[0264] The compounds of the present invention, their pharmaceutically acceptable salts, and pharmaceutically acceptable solvates of any of these entities may be administered alone, but are generally administered in mixture with appropriate pharmaceutical excipients, diluents, or carriers selected in relation to the intended route of administration and standard pharmaceutical practice.

[0265] Appropriate pharmaceutically acceptable excipients include, but are not limited to, any adjuvants, disintegrants, excipients, fluidizers, granulating binders, lubricants, sweeteners, preservatives, dyes, flavor enhancers, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers approved by the U.S. Food and Drug Administration for use in humans or livestock.

[0266] For example, the compounds of the present invention or their salts or solvates can be administered orally, buccally, or sublingually in the form of tablets, capsules (including soft gel capsules), suppositories, elixirs, liquids, or suspensions, which may contain flavorings or colorings, for immediate, delayed, modified, sustained, controlled-release, or pulsed delivery applications. The compounds of the present invention can also be administered through rapidly dispersible or rapidly dissolving dosage forms.

[0267] Such tablets may contain excipients such as crystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium hydrogen phosphate, and glycine; disintegrants such as starch (preferably corn, potato, or tapioca starch), sodium starch glycolate, croscarmellose sodium, and several complex silicates; and granulating binders such as polyvinylpyrrolidone, hydroxypropyl methylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin, and acacia. Furthermore, lubricants such as magnesium stearate, stearic acid, glyceryl behenate, and talc may be included.

[0268] Similar types of solid compositions can also be used as fillers in gelatin capsules. Preferred additives in this regard include lactose, starch, cellulose, lactose, or high molecular weight polyethylene glycol. In aqueous suspensions and / or elixirs, the compounds of the present invention can be used in combination with various sweeteners or flavorings, colorants or dyes, emulsifiers and / or suspending agents, and diluents such as water, ethanol, propylene glycol, glycerin, and combinations thereof.

[0269] Modified release and pulsed release formulations may contain additional excipients that act as release rate modifiers, such as those detailed for immediate release formulations, which are coated and / or included in the body of the device. Examples of release rate modifiers include, but are not limited to, hydroxypropyl methylcellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, cellulose acetate, polyethylene oxide, xanthan gum, carbomer, ammonia methacrylate copolymer, hydrogenated castor oil, carnauba wax, paraffin wax, cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, methacrylic acid copolymer, and mixtures thereof. Modified release and pulsed release formulations may contain one or a combination of release rate modifier excipients. The release rate modifier excipients may be present within the formulation, i.e., within the matrix, and / or on the formulation, i.e., on the surface or coating.

[0270] Rapidly dispersible or rapidly dissolving formulations (FDDFs) may contain the following materials: aspartame, acesulfame potassium, citric acid, croscarmellose sodium, crospovidone, diascorbic acid, ethyl acrylate, ethylcellulose, gelatin, hydroxypropyl methylcellulose, magnesium stearate, mannitol, methyl methacrylate, mint flavoring agent, polyethylene glycol, fumed silica, silicon dioxide, sodium starch glycolate, sodium stearyl fumarate, sorbitol, xylitol.

[0271] The compounds of the present invention may be administered parenterally, for example, intravenously or intra-arterially, or by infusion techniques. In such parenteral administration, they are best used in the form of a sterile aqueous solution that may contain sufficient salt or glucose to make a solution isotonic with other substances, such as blood. The aqueous solution should be buffered appropriately (preferably to pH 3-9) if necessary. The preparation of a suitable parenteral formulation under sterile conditions is readily achieved by standard formulation techniques well known to those skilled in the art.

[0272] Appropriately, the formulations of the present invention are optimized for the route of administration, such as oral or intravenous administration.

[0273] Administration may be a single dose during the course of treatment, either continuous or intermittent (e.g., divided doses at appropriate intervals). Methods for determining the most effective means and dosage are well known to those skilled in the art and vary depending on the formulation used for treatment, the purpose of treatment, the target cells being treated, and the subject being treated. Single or multiple doses may be administered at dose levels and in dose regimens selected by the treating physician, veterinarian, or clinician.

[0274] When more than one agent is administered, these agents may be provided by simultaneous or sequential administration. "Simultaneous" administration means that different agents are administered to the individual at the same time. This can be achieved as a single dose administered simultaneously via the same or different routes of administration. This can be done, for example, when one agent is administered by infusion or parenteral administration, and the other is administered orally during the course of the infusion or parenteral administration.

[0275] "Continuous" means that different agents are administered at different times, provided that the activity of the first agent is present and continues in the target when the second agent is administered. For example, an immunotherapy agent may be administered first to induce an immune response against the tumor antigen, and then a compound of formula (II), (III), (IV), (V), or (VI) or its derivatives, or its salts and solvates, may be administered to reduce immunosuppression at the tumor site. This continuous administration may be carried out via the same or different routes of administration. Preferably, the continuous administration is carried out so that the second agent of the two agents is administered within 48 hours, preferably within 24 hours, for example, within 12, 6, 4, 2, or 1 hour of the first agent. Preferably, one agent may be administered daily, and the second agent may be administered every 2, 3, 4, 5, 6, or 7 days.

[0276] Compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, can be administered multiple times, for example, two, three, four, five, or more times after administration of one or more immunotherapeutic agents or other anticancer agents. Administration of compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, can be continued for the duration following administration of immunotherapeutic agents or other anticancer agents. For example, treatment with compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, can be continued for at least one week, at least two weeks, at least three weeks, at least one month, or at least two months. Treatment with compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, can be interrupted and then resumed periodically. Treatment with compounds of formula (II), (III), (IV), (V), or (VI), or their salts and solvates, may be continued as long as complete tumor rejection needs to be achieved.

[0277] One or more immunotherapeutic agents or other anticancer agents may be administered multiple times, for example, two, three, four, five, or more times after administration of a compound of formula (II), (III), (IV), (V), or (VI) or its salts and solvates. Administration of immunotherapeutic agents or other anticancer agents may be continued for the duration following administration of a compound of formula (II), (III), (IV), (V), or (VI) or its salts and solvates. For example, treatment with immunotherapeutic agents or other anticancer agents may be continued for at least one week, at least two weeks, at least three weeks, at least one month, or at least two months. Treatment with immunotherapeutic agents or other anticancer agents may be continued as long as it is necessary to achieve complete tumor rejection.

[0278] Depending on the disorder being treated, the patient, and the route of administration, the composition may be administered in various doses. For example, a typical dose for adults may be 100 ng to 25 mg (preferably about 1 μg to about 10 mg) per kg of body weight per day.

[0279] When estimating initial doses for human subjects, advice can be drawn from studies in test animals. For example, when identifying a specific dose for mice, the initial test dose for humans may appropriately be approximately 0.5–2 × mg / kg values ​​given to mice.

[0280] Other forms Unless otherwise specified, the above includes well-known ions, salts, solvates, tautomers, and protected forms of these substituents. For example, reference to carboxylic acids (-RCOOH) means the anionic (carboxylate) form (-RCOO). - ), its salts or solvates, and the usual protected forms. Similarly, references to the amino group include the protonated form (-RN). + HR 1 R 2) include salts or solvates of amino groups, such as hydrochloride salts, and the usual protected forms of amino groups. Similarly, references to hydroxyl groups also include the anionic form (-O - ), its salts or solvates, and the usual protected forms.

[0281] Some compounds can exist in one or more specific geometric, optical, enantiomeric, diastereomeric, epimeric, atropisomeric, mesomeric, stereoisomeric, tautomeric, conformational, or anomeric forms, collectively referred to hereinafter as "isomers" (or "isomeric forms"), including but not limited to cis and trans forms; E and Z forms; c, t, and r forms; endo and exo forms; R, S, and meso forms; D and L forms; d and l forms; (+) and (-) forms; keto, enol, and enolate forms; syn and anti forms; synclinal and anticlinal forms; α and β forms; axial and equatorial forms; boat, chair, twisted, envelope, and half-chair forms; and combinations thereof.

[0282] Note that, except for the tautomeric forms described hereinafter, specifically excluded from the term "isomers" in this specification are structural (or constitutional) isomers (i.e., isomers that differ not merely in the position of atoms in space but in the bonds between atoms). For example, a reference to a methoxy group (-OCH3) should not be construed as a reference to its structural isomer, a hydroxymethyl group (-CH2OH).

[0283] References to classes of structures may include structural isomeric forms corresponding to that class (e.g., C 1-7 alkyl includes n-propyl and iso-propyl; butyl includes n-, iso-, sec-, and tert-butyl; methoxyphenyl includes ortho-, meta-, and para-methoxyphenyl).

[0284] The above exclusions do not apply to the tautomer forms, such as keto, enol, and enolate forms, in the case of the following tautomer pairs: keto / enol, imine / enamine, amide / iminoalcohol, amidine / amidine, nitroso / oxime, thioketone / enthiol, N-nitroso / hydroxyazo, and nitro / acinitro.

[0285] Please note that the term "isomer" specifically refers to compounds that have one or more isotopic substitutions. For example, H is 1 H, 2 H(D), and 3 It can take on any isotopic form, including H(T). C is 12 C, 13 C, and 14 It can take on any isotopic form, including C. O is, 16 O and 18 It can take on any isotopic form, including oxygen.

[0286] Unless otherwise specified, references to a particular compound include all such isomeric forms, including (complete or partial) racemic compounds and other mixtures thereof.

[0287] Methods for preparing (e.g., asymmetric synthesis) and separating (e.g., fractional crystallization and chromatographic means) such isomer morphologies are known in the art or can be readily obtained by methods taught herein or by adapting known methods in known ways.

[0288] Unless otherwise specified, references to specific compounds include, for example, their ions, salts, solvates, and protected forms, as described below.

[0289] In some embodiments, the compounds of the Disclosure, their salts, and solvates include pharmaceutically acceptable salts of the compounds of the Disclosure.

[0290] The compounds of this disclosure, including those specifically named above, can form pharmaceutically acceptable complexes, salts, solvates, and hydrates. Examples of these salts include non-toxic acid addition salts (including diacides) and base salts.

[0291] The compound is cationic, or has a functional group that can be cationic (for example, -NH2 is -NH3) +If (this is possible), the acid addition salt can be formed with a suitable anion. Examples of suitable inorganic anions include, but are not limited to, those derived from the following inorganic acids: hydrochloric acid, nitric acid, nitrite, phosphoric acid, sulfuric acid, sulfurous acid, hydrobromic acid, hydroiodic acid, hydrofluoric acid, phosphoric acid, and phosphorous acid. Examples of suitable organic anions include, but are not limited to, those derived from the following organic acids: 2-acetyloxybenzoic acid, acetic acid, ascorbic acid, aspartic acid, benzoic acid, camphor sulfonic acid, cinnamic acid, citric acid, edetic acid, ethanedisulfonic acid, ethanesulfonic acid, fumaric acid, glucheptonic acid, gluconic acid, glutamic acid, glycolic acid, hydroxymaleic acid, hydroxynaphthalenecarboxylic acid, isethionic acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, methanesulfonic acid, mucoic acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, pantothenic acid, phenylacetic acid, phenylsulfonic acid, propionic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, tartaric acid, toluenesulfonic acid, and valeric acid. Suitable polymer organic anions include, but are not limited to, those derived from polymer acids such as tannic acid and carboxymethylcellulose. Examples of such salts include acetate, adipine, aspartate, benzoate, besilate, bicarbonate, carbonate, bisulfate, sulfate, borate, camusylate, citrate, cyclamate, edicylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride, hydrobromide, hydroiodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfonate, naphthylate, 2-napsylate, nicotinate, nitrate, orotinate, oxalate, palmitate, pamoate, phosphate, hydrogen phosphate, dihydrogen phosphate, pyroglutamate, saccharinate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, and xinafoate.

[0292] For example, a compound is anionic, or has a functional group that can be anionic (for example, -RCOOH is -RCOO - If (this is possible), the base salt can form a suitable cation. Examples of suitable inorganic cations include, but are not limited to, metal cations such as alkali metal or alkaline earth metal cations, ammonium and substituted ammonium cations, and amines. An example of a suitable metal cation is sodium (Na + ), potassium (K + ), magnesium (Mg 2+ ), calcium (Ca 2+ ), zinc (Zn 2+ ), and aluminum (Al 3+ Examples of suitable organic cations include the ammonium ion (i.e., NH4). + ) and substituted ammonium ions (e.g., NH3R + NH2R2 + NHR3 + NR4 + Examples of suitable substituted ammonium ions include, but are not limited to, ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, and tromethamine, as well as those derived from amino acids such as lysine and arginine. A common example of a quaternary ammonium ion is N(CH3)4. +Appropriate amines include arginine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethylamine, diethanolamine, dicyclohexylamine, ethylenediamine, glycine, lysine, N-methylglucamine, olamine, 2-amino-2-hydroxymethylpropane-1,3-diol, and procaine. For a discussion of useful acid addition salts and base salts, see SM Berge et al., J. Pharm. Sci. (1977) 66:1-19. See also Stahl and Wermuth, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (2011).

[0293] Pharmaceutically acceptable salts can be prepared using a variety of methods. For example, a desired salt can be obtained by reacting the compounds of the disclosed herein with a suitable acid or base. A precursor of the compounds of the disclosed herein can also be reacted with an acid or base to remove acid- or base-unstable protecting groups, or to open the lactone or lactam groups of the precursor. Furthermore, a salt of a compound of the disclosed herein can be converted to another salt by treatment with a suitable acid or base or contact with an ion-exchange resin. After the reaction, the salt can then be isolated and recovered by filtration if it precipitates from the solution, or by evaporation. The degree of ionization of the salt can range from fully ionized to nearly non-ionized.

[0294] The preparation, purification, and / or handling of the corresponding solvates of the active compound may be convenient or desirable. The term "solvate" refers to a molecular complex containing the compound and one or more pharmaceutically acceptable solvent molecules (e.g., EtOH). The term "hydrate" refers to a solvate in which the solvent is water. Examples of pharmaceutically acceptable solvates include those in which the solvent is isotope-substituted (e.g., D2O, acetone-d6, DMSO-d6).

[0295] The currently accepted classification system for solvates and hydrates of organic compounds distinguishes between isolation sites, channels, and metal ion-coordinated solvates and hydrates. See, for example, KR Morris (HG Brittain ed.) Polymorphism in Pharmaceutical Solids (1995). Isolation site solvates and hydrates are those in which solvent (e.g., water) molecules are isolated from direct contact with each other by intermediary molecules of the organic compound. In channel solvates, solvent molecules reside in lattice channels adjacent to other solvent molecules. In metal ion-coordinated solvates, solvent molecules are bound to metal ions.

[0296] When the solvent or water is tightly bound, the complex exhibits a clearly defined stoichiometry regardless of humidity. However, when the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water or solvent content depends on humidity and dry conditions. In such cases, non-stoichiometry is typically observed.

[0297] Synthesis Strategy The method of the present invention may include one or more of the techniques described later. Some of the schemes and examples may omit details of common reactions, including oxidation, reduction, separation techniques (extraction, evaporation, precipitation, chromatography, filtration, tritulation, crystallization, etc.), and analytical procedures, which are known to those skilled in the art of organic chemistry. Details of such reactions and techniques can be found in several specialized books, including Richard Larock, *Comprehensive Organic Transformations*, *A Guide to Functional Group Preparations*, 2nd Ed (2010), and a multi-volume series edited by Michael B. Smith et al. (*Michael B. Smith and others*, *Compendium of Organic Synthetic Methods* (1974 et seq.)). Starting materials and reagents can be obtained from commercial suppliers or prepared using methods described in the literature. Some reaction schemes may omit small amounts of products resulting from chemical transformations (e.g., alcohols from ester hydrolysis, CO2 from diacid decarboxylation, etc.). Furthermore, in some cases, reaction intermediates can be used in subsequent steps (i.e., in situ) without isolation or purification.

[0298] In some of the following reaction schemes and examples, several compounds can be prepared using protecting groups to prevent undesirable chemical reactions at other reaction sites. Protecting groups can also be used to improve solubility or, in other cases, to modify the physical properties of compounds. For a discussion of protecting group strategies, a description of materials and methods for incorporating and removing protecting groups, and a compilation of useful protecting groups for common functional groups, including amines, carboxylic acids, alcohols, ketones, and aldehydes, see TW Greene and PG Wuts, Protecting Groups in Organic Chemistry, 4th Edition, (2006) and P. Kocienski, Protective Groups, 3rd Edition (2005).

[0299] In general, some reactions can be beneficial by using an excess of one or more reactants, but the chemical transformations described throughout this specification can be carried out using substantially stoichiometric amounts of reactants. Furthermore, while many of the reactions disclosed throughout this specification can be carried out at approximately room temperature (RT) and ambient pressure, some reactions can be carried out at high pressure, or at higher temperatures (e.g., under reflux) or lower temperatures (e.g., -78°C to 0°C), depending on the reaction kinetics, yield, etc. In this disclosure, references to stoichiometric ranges, temperature ranges, pH ranges, etc., include the indicated endpoints, whether the word “range” is explicitly used or not.

[0300] In many chemical transformations, one or more miscible solvents can be used, which can affect the reaction rate and yield. Depending on the properties of the reactants, one or more solvents may be protic polar solvents (including water), aprotic polar solvents, nonpolar solvents, or any combination thereof. Typical solvents include saturated aliphatic hydrocarbons (e.g., n-pentane, n-hexane, n-heptane, n-octane); aromatic hydrocarbons (e.g., benzene, toluene, xylene); halogenated hydrocarbons (e.g., methylene chloride, chloroform, carbon tetrachloride); and aliphatic alcohols (e.g., methanol, ethanol, propan-1-ol, propan-2-ol, butan-1-ol, 2-methyl-propan-1-ol, butan-2-ol, 2-methyl-propan-2-ol, pentan-1-ol, 3-methyl-butan-1-ol, hexane-1-ol, 2-methoxyethanol, 2-ethoxyethanol, 2-butoxyethanol, 2-(2-methoxyethoxy)ethanol, 2-(2-ethoxyethoxy)ethanol, 2-(2-butoxyethoxy)-ethanol Examples include ethanol; ethers (e.g., diethyl ether, di-isopropyl ether, dibutyl ether, 1,2-dimethoxyethane, 1,2-diethoxyethane, 1-methoxy-2-(2-methoxyethoxy)ethane, 1-ethoxy-2-(2-ethoxyethoxy)ethane, tetrahydrofuran, 1,4-dioxane); ketones (e.g., acetone, methyl ethyl ketone); esters (methyl acetate, ethyl acetate); nitrogen-containing solvents (e.g., formamide, N,N-dimethylformamide, acetonitrile, N-methylpyrrolidone, pyridine, quinoline, nitrobenzene); sulfur-containing solvents (e.g., carbon disulfide, dimethyl sulfoxide, tetrahydrothiophene-1,1-dioxide); and phosphorus-containing solvents (e.g., HMPA, hexamethylphosphoramide).

[0301] Further specific preferred embodiments are described in the attached independent and dependent claims. Features of the dependent claims may be combined with features of the independent claims as appropriate, and may be combinations other than those expressly described in the claims. [Examples]

[0302] Surprisingly, the inventors discovered that the phosphate KCL-HO-1i exhibited unexpected in vivo anticancer activity and demonstrated superiority over commercially available SnMP dichlorides in in vivo studies (see Section 2). KCL-HO-1i also appears to have a superior solubility profile compared to commercially available SnMPs.

[0303] The synthesis of KCl-HO-1i from SnMP is described below. Generally, SnMP can be brought into contact with a phosphate source at a predetermined temperature for a predetermined time, or MP can be brought into contact with a tin source and a phosphate source at a predetermined temperature for a predetermined time.

[0304] 1) Synthesis of KCL-HO-1i [ka] JPEG2026509438000084.jpg7170 Scheme 2. Synthesis of KCl-HO-1i using tin pyrophosphate

[0305] 2) KCL-HO-1i as an HO inhibitor with antitumor efficacy The inventors explored KCL-HO-1i in preclinical trials and demonstrated its superiority over dichloride SnMP (shown in Figures 3, 4, 5, 7, and 9). Administration of KCL-HO-1i to tumor mice with MMTV-PyMT demonstrated superior antitumor efficacy in combination with 5-FU chemotherapy (Figure 4). Remarkably, on day 8 after the start of treatment, KCL-HO-1i eradicated all tumor burdens in SnMP-treated animals, which still exhibited residual disease (Figure 5). Therefore, KCL-HO-1i, when used in combination with 5-FU, provides a measurable improvement in the antitumor immune response compared to standard commercially available SnMP (Figure 5).

[0306] Currently, MP is synthesized using animal-derived starting materials, specifically hemin.

[0307] There is a need for a synthetic route that improves the efficiency and scale of production, addresses ethical and religious concerns, and is vegan-friendly by not containing animal products. Surprisingly, the inventors provide a method for synthesizing MP using a method that enables large-scale production of MP without using animal products.

[0308] 3) Large-scale production of synthetic KCl-HO-1i The synthesis route illustrated in Scheme 3 below is a possible route to KCL-HO-1i that does not require animal starting materials.

[0309] SnMP produced using the synthetic pathway according to the present invention is active in delivering antitumor efficacy in the MMTV-PyMT mouse model of cancer when used in combination with chemotherapy 5-FU (see Figure 6). [ka] JPEG2026509438000086.jpg7170 Scheme 3. Optimal synthesis of large-scale synthetic production of sparrow mesoporphyrin phosphate (KCL-HO-1i) according to the present invention

[0310] KCL-HO-1i can be synthesized from MP produced in a large-scale synthesis in step 4 or from SnMP produced in step 5 by contacting MP or SnMP with a tin source and a phosphate source (for example, tin pyrophosphate as described in 1 above).

[0311] Stage 1 [ka] Pyrrole A(1) (200 g), pyrrole B(2) (309 g), and methanol (3.99 L) were placed in a reactor under nitrogen, and the contents were heated and refluxed. A methanol (10 mL) solution of p-toluenesulfonic acid monohydrate (1.21 g) was added over 3 minutes. The reaction mixture was stirred under reflux for 32 minutes, and then cooled to 0°C over 2 hours and 30 minutes on a controlled gradient. Spontaneous crystallization occurred at 38-36°C. At 2°C, the slurry was discharged into a filter for deliquency, and the container was rinsed with methanol (2 times × 500 mL) cooled to below 5°C. The cake was completely deliqued, excavated onto a drying tray, and vacuum-dried at 40°C for 8 hours to obtain product (3) (286.8 g, yield 90.7%).

[0312] Stage 2 [ka] Compound (3) (210.0 g), tetrahydrofuran (1050 mL), and triethylamine (147 mL) were placed in a flask, and the contents were stirred until a solution was formed. Alumina-supported 5% palladium (42.0 g) was placed in the flask, and hydrogen was dispersed in the stirred mixture at 500 mL / min for 52 minutes, during which time the temperature of the flask contents was raised from 17°C to 37°C. After this time, the reaction temperature began to decrease, and the gas flow discharged from the batch increased significantly by visual observation. After a reaction time of 60 minutes, the mixture was passed through a 25 mm deep Celite bed and placed in a 3 L pear-shaped flask. The filter bed was washed with tetrahydrofuran (4 times × 100 mL). The filtrate and washings were combined and concentrated under reduced pressure to obtain a wet grayish-white solid. This was vacuum-dried at 40°C for 20 hours to obtain a dry intermediate (174.2 g, yield 98.6%), which was used without purification. 1 Identity confirmed by 1H NMR.

[0313] Stage 3 [ka] Para-toluenesulfonic acid monohydrate (372.4 g), chloroform (10.61 L), and methanol (2.12 L) were placed in a flask and placed under a nitrogen atmosphere. A 5:1 chloroform / methanol (1.50 L) solution of the TEA salt (174.1 g) and dialdehyde (157.6 g) of the crude compound (26) from step 2 was added via an HPLC pump over 16 hours and 30 minutes. A 5:1 chloroform / methanol solution (40 mL x 3 times) was added as a rinse solution for the container and lines. The mixture was stirred for 1 hour. A solution of zinc acetate dihydrate (172.4 g) and methanol (2.50 L) was added. Air was dispersed in the mixture for 6 hours. The mixture was stirred for approximately 10 hours in an open atmosphere. The mixture was washed with deionized water (2 x 2.50 L), then with 20% w / w aqueous potassium carbonate (2.10 L, 2.5 kg), and finally with water (2.50 L). The chlorinated solution was concentrated under reduced pressure to obtain a dark brown / purple oily mass. This was vacuum-dried at 40°C for 20 hours to obtain a dry intermediate (21) (255.6 g, 99% yield), which was used without further purification or analysis.

[0314] Crude zinc porphyrin complex (21) (511.7 g) and a 5% v / v sulfuric acid methanol (2.50 L) solution were placed in a flask and stirred until the solids dissolved. After stirring for 4 hours, the mixture was transferred to a 20 L container and diluted with dichloromethane (5.00 L). The mixture was washed with deionized water (2.50 L). The aqueous layer was back-extracted with dichloromethane (2 × 0.5 L). The chlorinated phases were combined and washed with deionized water (2.50 L), then with 20% w / w aqueous potassium carbonate (2.10 L, 2.5 kg), and further washed with water (2.50 L). The chlorinated solution was concentrated under reduced pressure to obtain a dark brown / black oily mass. This was vacuum-dried at 40°C for 20 hours to obtain the dry crude step 3 product (6) (490.1 g). [ka] The crude Step 3 product (480 g) was dissolved in dichloromethane (approximately 3.5 L) and loaded onto a 5 kg HP-Sphere silica cartridge equilibrated with dichloromethane in a Biotage Flash 150 L system. The cartridge was eluted with dichloromethane (40 L) at approximately 800 mL / min, followed by 0.5% methanol in dichloromethane (130 L), and a fraction of approximately 10 L was collected. Elution was continued with 0.5% methanol in dichloromethane (30 L), and fractions of approximately 5 L each were collected six more times. The desired fractions were combined and concentrated under reduced pressure to obtain the Step 3 product as a brown / purple solid (214.3 g, yield 44.3%).

[0315] Stage 4 [ka] Compound (6) (85.0 g) from step 3, tetrabutylammonium chloride (850 mg), dichloromethane (1.70 L), methanol (1.70 L), and 2M aqueous sodium hydroxide (268 mL) were placed in a flask, and the contents were heated and refluxed (45°C). The mixture was stirred under reflux for 4 hours, after which the reaction was completed as determined by HPLC. The mixture was allowed to cool to room temperature. Concentrated hydrochloric acid (56.2 g) was added dropwise over 10 minutes. After stirring the mixture for 1 hour, it was discharged into a filter and dehydrated, and the container was rinsed with a 1:1 mixture of dichloromethane and methanol (2 times × 150 mL), then with water (2 times × 100 mL) into the filter. The filtration cake was completely dehydrated and excavated.

[0316] The moist product cake, deionized water (300 mL), and methanol (300 mL) were placed in a flask, the contents were heated to 70°C and stirred for 30 minutes, and then cooled to room temperature. The slurry was discharged through a filter and dehydrated, and the container was rinsed through a filter with a 1:1 mixture of methanol and deionized water (twice × 150 mL), then methanol (100 mL). The filtered cake was completely dehydrated, excavated onto a drying tray, and vacuum-dried at 45°C for 20 hours to obtain the Step 4 product as a brown / purple powder (71.6 g). The liquids from both filters were concentrated under reduced pressure to obtain a slurry of approximately 600 mL in volume. This slurry was discharged through a filter and dehydrated. The filtered cake was washed with deionized water (twice × 100 mL). The filtered cake was completely dehydrated, excavated onto a drying tray, and vacuum-dried at 45°C for 20 hours to obtain the second harvest of the Step 4 product (7) as a brown / purple powder (7.2 g). The Step 4 product (7) was combined to obtain a brown / purple powder (78.8 g, yield 97.3%).

[0317] Stage 5 [ka] 200 mg of mesoporphyrin IX (0.35 mmol, 1 equivalent) and 576 mg of tin(II) pyrophosphate (1.4 mmol, 4 equivalents) were added to a 50 ml round-bottom flask lined with tin foil. 5 ml of glacial acetic acid was added, and nitrogen gas was passed through the flask. The mixture was then stirred under reflux at 115°C. Once the temperature stabilized, the reaction was opened to air to introduce oxygen into the system, and the reaction was left to stand for 24 hours. The mixture was then cooled to room temperature, quenched with 4 ml of HPLC-grade water, filtered, and air-dried for 10 minutes. The filtered solid was placed in another 50 ml flask with 3 ml of HPLC-grade water and 0.48 ml of concentrated HCl, and stirred at 90°C for 30 minutes. The mixture was cooled to room temperature, filtered, and washed with cold water. After air drying for 30 minutes, 125 mg of the product (KCL-HO-1i) was obtained as a red powder (40% isolated yield). 1H NMR(400MHz,DMSO-d6)δ10.36(s,1H), 10.13(s,1H), 10.09(d,J=4Hz,2H), 7.11(s,4H), 3.82~3.96(m,5H), 3.62(q,J=14Hz,4H), 3.17(d,J=12Hz,12H), 2.64(t,J=6Hz,5H), 1.82(t,J=2Hz,16H), 1.25(t,J=6Hz,8H).

[0318] Alternative Stage 5 This general reaction scheme can also be applied to the preparation of SnMP by using the following alternative step 5. [ka] 101.7 g of tin(II) oxide and 2.50 L of acetic acid were placed in a flask, and the contents were heated to 65°C. While maintaining the temperature of the mixture in the flask at 60-70°C, the solution of step 4(7) (115 g) in the mixture of formic acid (590 mL) and concentrated hydrochloric acid (43.2 g) was added dropwise over 3 hours and 40 minutes. Formic acid (20 mL x 3 times) was added as line rinses. The formation of a deep red precipitate became apparent after approximately 2 hours and 45 minutes (approximately 75% by addition). The mixture was stirred at 63-68°C for 17 hours and 25 minutes. 1.05 L of deionized water was added over 20 minutes. The mixture was cooled to 20°C over 3 hours and stirred for a further 2 hours. The slurry was discharged into a filter for deliquescence, and the container was rinsed with deionized water (100 mL x 2 times) into the filter. The filtration cake was dehydrated, excavated, and returned to the flask. 1.75 L of 1 M aqueous hydrochloric acid was added to the flask, and the contents were heated to 85°C. The mixture was stirred at 85-90°C for 80 minutes, after which the slurry was discharged into a filter for dehydration, and the container was rinsed with water (250 mL twice) through the filter. The filtration cake was completely dehydrated, excavated onto a drying tray, and vacuum-dried at 40°C for 40 hours to obtain sproutsoporphyrin as a deep red powder (135.7 g, yield 88.7%).

[0319] Inductively coupled plasma mass spectrometry (ICP-MS) comparisons were performed between SnMP (tin dichloride salt of mesoporphyrin IX) and two KCl-HO-1i samples (tin phosphate salt of mesoporphyrin IX) to enable elemental analysis of the tin content of these compounds. [Table 1] Therefore, this elemental analysis data indicates that KCL-HO1i has, on average, three times more Sn than SnMP.

[0320] This general reaction scheme can also be applied to the synthesis of mesophorphorin IX tartrate (TA-191-149) by using the following alternative step 5. [ka] 200 mg of mesoporphyrin IX (0.35 mmol, 1 equivalent) and 372.5 mg of tin(II) tartrate (1.4 mmol, 4 equivalents) were added to a 50 ml round-bottom flask lined with tin foil. 5 ml of glacial acetic acid was added, and nitrogen gas was passed through the flask twice. The mixture was then stirred under reflux at 120°C. Once the temperature stabilized, the reaction was opened to air to introduce oxygen into the system, and the reaction was left to stand for 24 hours. The mixture was then cooled to room temperature, quenched with 4 ml of HPLC-grade water, filtered, and air-dried for 10 minutes. The filtered solid was placed in another 50 ml flask with 3.5 ml of 1 M HCl and stirred at 90°C for 30 minutes. The mixture was cooled to room temperature, filtered, and washed with cold water. After air-drying for 30 minutes, 125 mg of the product was obtained as a brick-red powder (isolation yield 32%). Melting point: 287.2℃; 1 H NMR(400MHz, methanol-d4)d 11.21(br.s.,1H), 11.01(br.s.,1H), 10.97(br.s.,2H), 4.45(d,J=7.06Hz,5H), 3.95(s,7H), 3.98(s,8H), 3.47(br.s.,5H), 2.65(br.s.,2H), 1.88~2.16(m,8H).

[0321] This general reaction scheme can also be applied to the synthesis of mesophorphorin IX oxalate (TA-191-151) by using the following alternative step 5. [ka] 200 mg of mesoporphyrin IX (0.35 mmol, 1 equivalent) and 290 mg of tin(II) oxalate (1.4 mmol, 4 equivalents) were added to a 50 ml round-bottom flask lined with tin foil. 5 ml of glacial acetic acid was added, and nitrogen gas was passed through the flask twice. The mixture was then stirred under reflux at 120°C. Once the temperature stabilized, the reaction was opened to air to introduce oxygen into the system, and the reaction was then left to stand for 24 hours. The mixture was then cooled to room temperature, quenched with 4 ml of HPLC-grade water, filtered, and air-dried for 10 minutes. The filtered solid was placed in another 50 ml flask with 3.5 ml of 1 M HCl and stirred at 90°C for 30 minutes. The mixture was cooled to room temperature, filtered, and washed with cold water. After air-drying for 30 minutes, 130 mg of the product was obtained as a brick-red powder (isolation yield 38%). Melting point: 263.2℃; 1 H NMR(400MHz, methanol-d4)d 11.08(d,J=8.34Hz,1H), 10.89(d,J=6.69Hz,1H), 10.85(d,J=5.41Hz,2H), 4.19~4.3 9(m,5H), 3.67~3.93(m,14H), 3.27~3.47(m,5H), 3.23(s,1H), 1.91(q,J=7.40Hz,7H)

[0322] This general reaction scheme can also be applied to the synthesis of mesophorphorin IX mesylate (TA-191-153) by using the following alternative step 5. [ka] 200 mg of mesoporphyrin IX (0.35 mmol, 1 equivalent) and 290 mg of tin(II) methylsulfonate (1.4 mmol, 4 equivalents) were added to a 50 ml round-bottom flask lined with tin foil. 5 ml of glacial acetic acid was added, and nitrogen gas was passed through the flask twice. The mixture was then stirred under reflux at 120°C. Once the temperature stabilized, the reaction was opened to air to introduce oxygen into the system, and the reaction was left to stand for 24 hours. The mixture was then cooled to room temperature, quenched with 4 ml of HPLC-grade water, filtered, and air-dried for 10 minutes. The filtered solid was placed in another 50 ml flask with 3.5 ml of 1 M HCl and stirred at 90°C for 30 minutes. The mixture was cooled to room temperature, filtered, and washed with cold water. After air-drying for 30 minutes, 72 mg of the product was obtained as a red powder (isolation yield 24%). Melting point: 257.8℃; 1 H NMR(400MHz, methanol-d4)d 11.25(d,J=16.05Hz,1H), 10.94~11.14(m,3H), 4.71~4.80(m,4H), 4.39~4.53(m,4 H), 3.91~4.04(m,12H), 3.47(t,J=7.24Hz,4H), 2.70(s,6H), 2.03(t,J=7.57Hz,6H)

[0323] 4) Synthesis of pyrrole A [ka] The selective reduction step (iii) can be achieved by (a) Wolff-Kishner reduction using N2H2, KOH; (b) Clemmensen reduction using Zn-Hg, HCl; (c) catalytic hydrogenation using Pt-C, H2 under pressure; or (d) using TsNHNH2, then NaBH3CN.

[0324] 4) Synthesis of pyrrole B [ka] JPEG2026509438000101.jpg9170 (a) KOH, benzyltrimethylammonium bromide, CHCl3; (b) pen-3-in-2-one, 5 mol% Cu catalyst (as Cu2O), 20 mol% PPh3, dioxane, 6-8 hours at 100°C (see reference 19 below); (c) (i) Wolff-Kishner reduction using N2H2, KOH; (ii) Clemmensen reduction using Zn-Hg, HCl; (iii) Catalytic hydrogenation using Pt-C, H2 under pressure; or (IV) Selective reduction using TsNHNH2, then NaBH3CN.

[0325] 5) Synthesis of Dialdehydes [ka] For further details regarding step (v), please refer to reference 20 below.

[0326] Pharmacokinetic (PK) research In male ICRs, pharmacokinetic (PK) studies were conducted after oral administration (PO) of SnMP (CR) and KCL-HO-1i (NS) at doses of 38 and 44 mg / kg, respectively. Plasma samples were collected a predetermined time after PO administration. Two formulations containing SnMP(CR) and KCL-HO-1i 7(NS) were prepared as shown in Table 1 below. The administration volume was 9.5 mL / kg for SnMP(CR) and 11 mL / kg for KCL-HO-1i(NS).

[0327] SnMP(CR) preparation (4 mg / mL). 23.38 mg of SnMP(CR) powder is dissolved in 1.17 mL of 0.1 M sodium hydroxide (NaOH); after complete dissolution, 4.68 mL of 0.5 M sodium bicarbonate (NaHCO3) is added at pH 7 to achieve a final concentration of 4 mg / mL.

[0328] Preparation of KCl-HO-1i(NS) (4 mg / mL). 26.68 mg of KCl-HO-1i(NS) powder is dissolved in 1.334 mL of 0.1 M NaOH. After complete dissolution, 5.336 mL of 0.5 M NaHCO3 is added at pH 7 to achieve a final concentration of 4 mg / mL.

[0329] Male ICR mice weighing 25±5g were obtained by BioLasco Taiwan (under the license of Charles RIVer Laboratories). The animals were acclimatized for 3 days prior to use to ensure good health. All animals were maintained in a hygienic environment with controlled temperature (20–24°C), humidity (30%–70%), and a 12-hour light / dark cycle. They were given free access to sterile standard laboratory diet [MFG (Oriental Yeast Co., Ltd., Japan)] and autoclaved tap water. All aspects of this study, including enclosures, experiments, and animal handling, were conducted in accordance with the Guide for the Care and Use of Laboratory Animals: Eighth Edition (National Academy Press, Washington, DC, 2011) at the inventors' AAALAC-accredited laboratory animal facility. Animal care and use protocols were reviewed and approved by IACUC of Pharmacology Discovery Services Taiwan, Ltd.

[0330] The animal dosage design is described in Table 2 below.

[0331] Collection of plasma samples from mice (parallel) A fixed amount of blood (approximately 300 μL) was collected from euthanized mice by cardiac puncture using a tube coated with lithium heparin, gently mixed, and centrifuged at 2,500 × g for 15 minutes at 4°C within one hour of collection. Plasma samples were then collected and kept frozen at -70°C until further processing.

[0332] Quantitative bioanalysis (plasma samples) and pharmacokinetics Plasma samples were treated with methanol precipitation and analyzed by LC-MS / MS. The exposure levels (ng / mL) of the formulations [SnMP or KCl-HO-1i] in the plasma samples were then determined by LC-MS / MS. Figure 1 shows the SnMP exposure levels (ng / mL) in mouse plasma samples after SnMP (38 mg / kg, PO) administration, Figure 2 shows the levels for KCl-HO-1i, and Figure 3 shows a combined plot. [Table 2] (a) This is based on visual observation: S: Soluble; SS: Slightly soluble; I: Insoluble (suspended or precipitated) (b) Y: Maintain the mixture in a brown tube or vial, or cover it with aluminum foil. (c) RT: Prepare fresh and store at 20-25°C. 4°C: Prepare fresh and store in a refrigerator or keep on ice. Table 1: Formulation [Table 3] Table 2: Animal Dosage Design

[0333] In vivo antitumor effects of various single and combination therapies Mice with invasive breast cancer model, invasive mammary tumor (MMTV-PyM), received an intraperitoneal (ip) administration strategy offering individual treatments of KCL-HO-1i (25 μMol / kg / day), SnMP (25 μMol / kg / day), 5-FU (40 mg / kg / 4 days), gemcitabine (64 mg / kg / 7 days), or medium injection, or a combination dual treatment of (i) KCL-HO-1i or SnMP and (ii) 5-FU or gemcitabine (see Figures 4A and 4B). Treatment was initiated on day 0 and marked with vertical dashed lines in Figures 4C–4L. For each individual mouse tumor, tumor growth was monitored, tumor volume was calculated, and the results are shown as growth curves for different single treatments (see Figures 4C–4G) and combination treatments (see Figures 4H–4J). The combination of KCL-HO-1i and chemotherapy (5-FU or gemcitabine) provides durable antitumor efficacy, as shown in Figures 4K and 4L, where each line represents the mean (n=5 for each treatment group) (** P<0.01). Remarkably, KCL-HO-1i and 5-FU treatment eradicated all tumor burden by day 8 after the start of treatment, resulting in a measurable improvement in the antitumor immune response compared to standard commercially available SnMP and 5-FU treatment, in which animals still had residual disease (see Figure 5).

[0334] Bioavailability Plasma concentrations were measured using a three-step process including (1) (ip) injection or forced oral administration of SnMP (25 μMol / kg) or KCL-HO1i (25 μMol / kg); (2) blood collection; and (3) LC / MS / MS measurement. Figure 7(A) shows a schematic diagram illustrating this process. Male mice received intraperitoneal administration (see Figure 7(B) (left panel)) or oral administration (see Figure 7(B) (right panel)) of commercially available SnMP or KCL-HO1i. At 0.167, 0.5, 1, 2, 4, 6, 8, and 24 hours after oral administration of SnMP, a certain volume of blood was collected by cardiac puncture in a heparin-coated lithium tube. After centrifugation, plasma samples were collected and maintained at -70°C until processing. The exposure levels (ng / mL) of SnMP (blue curve), veSnMP, and KCL-HO1i (red curve) in plasma samples are shown in Figure 7(B) (left panel) for intraperitoneal administration and Figure 7(B) (right panel) for oral administration. These exposure levels were determined by liquid chromatography-tandem mass spectrometry (LC-MS / MS) (Figure 7A). The plasma concentrations of SnMP and KCL-HO1i in mouse plasma are shown as mean ± SEM (standard error of the mean) versus time. The bar graph represents the mean, and the dots represent individual data points from individual mice.

[0335] A bioavailability study was conducted to test KCL-HO1i against commercially available SnMPs using bioluminescence. luc / eGFP A schematic diagram illustrating the mouse model and the strategy followed for bioluminescence is shown in Figure 7(C). HO-1 luc / eGFP Mice were injected with D-luciferin and imaged at T0 and 24 hours after treatment. Organs were collected and imaged. HO-1 treated with commercially available SnMP or KCL-HO1i at T0 (upper panel) and 24 hours after systemic treatment (lower panel). luc / eGFPRepresentative bioluminescence images comparing relative luciferase expression between mice are shown in Figure 7(D). Relative quantification of luciferase expression (photons / second) in whole-body and tissues is shown in Figure 7(E). Illustrative diagrams of individual tissue and total HO-1 and HO-1 expression changes in mice were obtained by normalizing to the medium group. Unexpectedly, KCL-HO-1i does not induce HO-1 expression, which is considered an undesirable side effect of commercially available SnMPs (Figures 7E, F). Figure 7(G) shows the evaluation of inhibition of HO-1 activity in rat microsomes using various doses of KCL-HO-1i and SnMP (biological replicates n=3) with concentrations of KCL-HO-1i and SnMP (μM) shown on a logarithmic scale. (*P<0.05, **P<0.01, ****P<0.001). These results demonstrate that KCL-HO-1i exhibits better availability in vivo than commercially available SnMPs.

[0336] In vivo antitumor effect of oral KCl-HO-1i in combination with gemcitabine Mice with spontaneously occurring tumors (MMTV-PyM), an invasive breast cancer model, were administered gemcitabine (intraperitoneal administration) in combination with KCL-HO-1i (oral administration), as shown in Figure 8A. Tumor measurement began on day 0 (cohort of 2 mice). Tumor growth was monitored, and tumor volume was calculated for each mouse (shown in Figure 8B, where each line represents an individual mouse and tumor). The combination of KCL-HO-1i and gemcitabine suppressed tumor growth.

[0337] Study of CD8 T cell infiltration into tumors in MMTV-PyMT mice 500~750mm 3MMTV-PyMT spontaneous tumor mice with tumors reaching a certain stage were used. Mice were administered equivalent doses of commercially available SnMP (cSnMP) or KCL-HO-1i on days 0 and 1, and the tumors were then harvested on day 2 (36 hours after the first injection). The tumors were enzymatically digested to release single cells, and the percentage of CD8+CD3+CD45+ T cells was assessed by flow cytometry. CD8 T cell infiltration into tumors of MMTV-PyMT mice treated with the appropriate medium, commercially available SnMP (cSnMP), or KCL-HO-1i is shown as bar graphs in Figure 9, where dots represent individual tumors and mice. *** P<0.001.

[0338] Efficacy of immunodepletion anti-CD8α antibodies to the in vivo antitumor effect of KCl-HO-1i and gemcitabine combination therapy Mice with spontaneously occurring tumors (MMTV-PyM) were treated with non-immune IgG or immunodepleted anti-CD8α antibody. Each antibody treatment was administered to the mice as an initial dose on day 2, followed by subsequent doses every 4 days. Some of these mice were also given KCL-HO-1i (25 μMol / kg / day) and gemcitabine (64 mg / kg / 7 days), starting on day 0. Tumor growth was monitored for each mouse, and tumor volume was calculated. Tumor growth curves are shown in Figures 10B-10E. In Figures 10B, 10C, and 10D, dots represent individual tumors and mice. Figure 10E is a line graph where each line represents each treatment and the average of the bars. In Figure 10, the dashed black line marks the start of treatment (day 0), and the red line marks 250 mm. 3 The result was marked. ** P<0.01. These results, particularly those of KCL-HO-1i / gemcitabine / anti-CD8α antibody compared to KCL-HO-1i / gemcitabine (see Figure 10E), demonstrate that the antitumor effects of KCL-HO-1i and gemcitabine are immunotherapeutic.

[0339] Investigation of T cell infiltration into tumors and T cell effector function Mice with established MMTV-PyM (MMTV-PyM) were treated with KCL-HO-1i (25 μMol / kg / day) and / or 5-FU (40 mg / kg / 4 days) or gemcitabine (64 mg / kg / 7 days) or a medium. After analyzing the tumors, treatment was initiated and re-analyzed 36 hours after the start of treatment. Tumor growth over the 36-hour treatment period is shown in the line graphs in Figure 11B for each treatment. The tumor microenvironment population was evaluated by removing and collecting the treated MMTV-PyM-derived tumors and analyzing their stromal composition by flow cytometry. There were no significant changes in the stromal population of most cells (see Figure 11C). CD8 + From T cell studies, KCL-HO-1i is CD8 + It was found that KCL-HO1i induces the influx of T cells into the tumor microenvironment, and this was promoted by the combined use of KCL-HO1i with chemotherapy (see bar graph Figure 11D). KCL-HO-1i treatment promotes CD8 + T-cell effector function was improved, as assessed by its IFN-γ expression, and this improvement was amplified by the combination of KCL-HO-1i and chemotherapy (see bar graph Figure 11E). There is evidence of synergistic effects between KCL-HO-1i and the chemotherapeutic agents tested. In contrast to KCL-HO-1i, SnMP, using equivalent doses and regimens to KCL-HO-1i, increased CD8 in the tumor microenvironment in MMTV-PyMT mice. + It was found not to induce T cell infiltration (see bar graph Figure 11F). Flow cytometry involves investigating and quantifying desired populations using fluorescent cell sorting (FACS) with gating. FACS-gated live cells showing CD44 and CD62L expression in medium-treated mice (7AAD) - ), CD45 + CD3 + CD8 + A representative dot plot of T cells is shown in Figure 11G, and the histogram shows CD8 across different treatment groups. + This represents a subpopulation of T cells (shown in Figure 11G), as shown in Figure 11H.

[0340] The bar graph shows the mean, and the dots represent individual data points from individual tumors and mice. The line graph shows the mean and SEM. * P<0.05, ** P<0.01.

[0341] Investigation using bulk RNA sequencing analysis Mice with established MMTV-PyMT tumors were intraperitoneally administered KCL-HO-1i (25 μMol / kg / day) and / or 5-FU (40 mg / kg) or gemcitabine (64 mg / kg) or the media. Tumor tissue was removed and collected as schematicly shown in Figure 12A, and analyzed at 36 hours by bulk RNA-seq analysis (cohort of mice and tumors n=5). From this analysis, Venn diagrams were plotted showing all upregulatory DEGs for each treatment of the media and their intercepts (see Figure 12B), and KCL-HO-1i treatment-related upregulatory DEGs (see Figure 12C). Heatmaps were also created for common upregulatory DEGs (419 genes) hierarchically clustered across treatment groups, which are secretory genes (91 genes) across the entire treatment. The scale ranges from low to high maximum change from the media-treated tumor (see Figure 12D). From this analysis, we also plotted Venn diagrams showing all upregulated DEGs in treatments including chemotherapy (5-FU or gemcitabine, alone or in combination with KCL-HO-1i) against the medium (see Figure 12E). Venn diagrams showing chemokine and upregulated DEG-related dual therapy treatments (KCL-HO-1i / 5-FU or KCL-HO-1i / gemcitabine) and the overlap between these groups against the medium (see Figure 12F). We also created heatmaps for chemokine and cytokine upregulated DEGs, hierarchically clustered across dual therapy groups throughout the treatment. The scale ranges from low to high, representing the maximum change from the medium-treated tumor (see Figure 12G).

[0342] In vivo treatment with oral KCl-HO-1i and gemcitabine, and analysis of animal physical characteristics and blood. Mice with established MMTV-PyMT tumors were orally administered KCL-HO-1i (25 μMol / kg / day) and / or intraperitoneally administered gemcitabine (64 mg / kg / 7 days) or a solvent (see Figure 13A). Figure 13B shows a line graph of tumor growth, where each line represents the average for mice that received each treatment. This shows that tumor growth was controlled by the combined use of KCL-HO-1i and gemcitabine. The body weight of the mice was measured during treatment and it was found that they maintained a body weight close to their baseline weight (see Figure 13C). On day 21 after treatment, blood samples were collected and plasma was separated from the blood by centrifugation. Serum liver aspartate aminotransferase (AST) and alanine aminotransferase (ALT) enzymes were evaluated by enzyme-linked immunosorbent assay (ELISA), and the AST / ALT ratio was evaluated and shown in a histogram (see Figure 13D) (n=5 for each treatment group). This revealed that no significant changes were observed in any of the treatment groups compared to treatment with the medium (see Figure 13D). The composition of immune cells in the blood was measured (see Figure 13E), and the animals' physical characteristics were examined on day 21 by their behavioral responses to any signs of distress, and these were scored on a scale of 0 to 5 (see heatmap in Figure 13F). No signs of distress were observed in the animals, and these results, along with weight measurements and blood analysis, all indicate that these treatments were nontoxic at the tested doses.

[0343] The bar graph shows the mean, and the dots represent individual data points from individual tumors and mice. The line graph shows the mean and SEM. * P<0.05, ** P<0.01.

[0344] Histopathological evaluation of organs in mice treated with KCL-HO-1i and / or gemcitabine. Mice were administered KCL-HO-1i (25 μMol / kg / day, 5-day treatment and 2-day rest regimen) via forced oral delivery, and / or gemcitabine (64 mg / kg / 7 days) via ip delivery, or administered a medium in MMTV-PyMT mice with established tumors (n=5 in each treatment group). On day 21 after the start of treatment, the indicated tissues were collected for histological examination, and the tissue sections and tissues from each mouse were stained with hematoxylin and eosin (H&E) and scored by a pathologist. These scores were based on a non-linear semi-quantitative evaluation system from 0 to 5, where 0 indicates no significant change and 5 indicates that the entire organ or tissue was affected in each observation.

[0345] Histological examination of brain, heart, kidney, and liver tissues showed little to no significant changes in any of the treatments. However, when pathologists scored mononuclear inflammatory cell infiltration and multifocal (perivascular) lesions in lung tissue histological examination, mice treated with KCL-HO-1i and gemcitabine scored up to 3. Similarly, for the lungs, when pathologists scored mixed inflammatory cell infiltration and focal (alveolar) lesions, mice treated with KCL-HO-1i and gemcitabine scored up to 2. However, when scoring metastases in lung tissue histological examination and evaluating foam macrophage (alveolar) lesions, the lungs were scored 0.

[0346] A separate histopathological evaluation of mouse lungs was performed in tumor-free C57Bl / 6 mice treated with a dual therapy of KCL-HO-1i (25 μMol / kg / day) and gemcitabine (64 mg / kg / 7 days) or orally administered the medium for 21 days. At the end of the treatment, lungs were then resected from one cohort of mice, while the second cohort was left untreated for 30 days before sacrifice. Lung sections were stained with H&E and evaluated by a pathologist, and scored based on the same nonlinear semi-quantitative evaluation system. In the dual therapy, at the end of the treatment, histological examination of lung tissue again showed scores of up to 3 for mononuclear inflammatory cell infiltration, multifocal (perivascular) lesions, and scores of up to 2 for mixed inflammatory cell infiltration, focal (alveolar) lesions. However, 30 days after the end of treatment, the scores for these lesions decreased, and in some mice, a score of 0 was recorded in the histological examination of lung tissue.

[0347] These histopathological evaluations suggest that while some pulmonary toxicity is observed with the KCL-HO-1i and gemcitabine dual treatment, these issues begin to resolve after discontinuation of the treatment. The absence of metastasis after dual treatment indicates that KCL-HO-1i may have an anti-metastatic effect.

[0348] Further in vivo antitumor effects of oral KCl-HO-1i in combination with gemcitabine Mice (C57B1 / 6) with established subcutaneous sarcoma tumors (MN-MCA-1 cell line) were administered KCL-HO-1i (25 μMol / kg / day, orally) and / or intraperitoneally administered gemcitabine (64 mg / kg / 7 days) or a medium, as schematically shown in Figure 14A. Tumor measurements were started on day 0. Tumor growth was monitored and tumor volume was calculated for each mouse. The tumor growth curves for each treatment are shown in Figure 14B, where the line represents the mean for treated mice and the bar represents SEM. The combination of KCL-HO-1i and gemcitabine suppressed tumor growth in these subcutaneous sarcoma tumors. ** P < 0.01.

[0349] Anti-metastatic effect 4T1 mammary gland cancer cells were orthotopically transplanted into Balb / c mice (BioLasco Taiwan, under the license of Charles River Laboratories) for tumor formation. A total of 2.5 × 10¹⁶ cells were obtained from 100 μl of Roswell Park Memorial Institute medium (RPMI 1640 medium). 5 Individual cells were subcutaneously injected into the mammary fat pad of syngeneic female mice, and tumor growth was then monitored, as shown in Figure 15A. Twelve days after tumor injection, mice were intraperitoneally administered either KCL-HO-1i (25 μMol / kg) or the medium daily for 14 days. Twenty-six days after tumor injection, lungs were harvested, and metastatic nodules were counted in both the KCL-HO-1i and medium-treated groups, as shown in the histogram in Figure 15B. KCL-HO-1i showed an anti-metastatic effect in 4T1 mice compared to the medium-treated group (see Figure 15B). Dots represent individual mice. **P<0.01.** This model indicates that primary tumor growth is unaffected, and therefore, the absence of lung metastases observed in the KCL-HO-1i medium-treated group does not simply reflect a lower tumor burden.

[0350] (References) 1. HA Tawbi et al., Combined Nivolumab and Ipilimumab in Melanoma Metastatic to the Brain. The New England journal of medicine 379, 722-730 (2018). 2. M. Husnain et al., Complete response to ipilimumab and nivolumab therapy in a patient with extensive extrapulmonary high-grade small cell carcinoma of the pancreas and HIV infection. Journal for immunotherapy of cancer 6, 66 (2018). 3. C. Robert et al., Pembrolizumab versus Ipilimumab in Advanced Melanoma. The New England journal of medicine 372, 2521-2532 (2015). 4. J. Larkin et al., Combined Nivolumab and Ipilimumab or Monotherapy in Untreated Melanoma. The New England journal of medicine 373, 23-34 (2015). 5. T. Muliaditan et al., Repurposing tin mesoporphyrin as an immune checkpoint inhibitor shows therapeutic efficacy in preclinical models of cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, (2018). 6. R. Gozzelino et al., Mechanisms of cell protection by heme oxygenase-1. Annual review of pharmacology and toxicology 50, 323-354 (2010). 7. K. Minamoto et al., Reciprocal regulation of airway rejection by the inducible gas-forming enzymes heme oxygenase and nitric oxide synthase. The Journal of experimental medicine 202, 283-294 (2005). 8. S. Brouard et al., Carbon monoxide generated by heme oxygenase 1 suppresses endothelial cell apoptosis. The Journal of experimental medicine 192, 1015-1026 (2000). 9. X. Zhang et al., Carbon monoxide differentially modulates STAT1 and STAT3 and inhibits apoptosis via a phosphatidylinositol 3-kinase / Akt and p38 kinase-dependent STAT3 pathway during anoxia-reoxygenation injury. The Journal of biological chemistry 280, 8714-8721 (2005). 10. J. N. Arnold et al., Tumoral immune suppression by macrophages expressing fibroblast activation protein-alpha and heme oxygenase-1. Cancer immunology research 2, 121-126 (2014). 11. M. B. Smith, "March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure", Eight Edition, 2020, Wiley. 12. O. V. Larionov et al., Versatile Direct Synthesis of Oligosubstituted Pyrroles by Cycloaddition of α-Metallated Isocyanides to Acetylenes. Angew. Chem. Int. Ed., 44, 5664-5667 (2005,). 13. F. Ebstein et al., Proteasomes generate spliced epitopes by two different mechanisms and as efficiently as non-spliced epitopes. Sci. Rep. 6, 24032; DOI: 10.1038 / srep24032 (2016). 14. J. Liepe et al., A large fraction of HLA class I ligands are proteasome-generated spliced peptides. Science, 354, 354-358 (2016). 15. S. Chai et al., NeoSplice: a bioinformatics method for prediction of splice variant neoantigens. Bioinformatics Advances, 2, 1-10 https: / / doi.org / 10.1093 / bioadv / vbac032 (2022). 16. P. Khongorzul et al., Antibody-Drug Conjugates: A Comprehensive Review, Mol Cancer Res, 18, 3-19; DOI: 10.1158 / 1541-7786.MCR-19-0582 (2020) 17. J. T. Bulcha et al., Viral Vector Platforms Within the Gene Therapy Landscape. Signal Transduction and Targeted Therapy, 6, Article number: 53, https: / / doi.org / 10.1038 / s41392-021-00487-6 (2021). 18. S. Ghosh et al., Viral Vector Systems for Gene Therapy: A Comprehensive Literature Review of Progress and Biosafety Challenges. Applied Biosafety 25, 7-18, http: / / doi.org / 10.1177 / 1535676019899502 (2020). 19. S. Kamijo et al., Copper- or Phosphine-Catalyzed Reaction of Alkynes with Isocyanides. Regioselective Synthesis of Substituted Pyrroles Controlled by the Catalyst. J. Am. Chem. Soc. 127, 9260-9266, https: / / doi.org / 10.1021 / ja051875m (2005). 20. KM Smith et al., Novel porphyrins from copper(II)-mediated cyclizations of 1',8'-dimethyl-A,C-biladiene salts: mechanism of the cyclization reaction. J. Org. Chem. 50, 2073-2080, https: / / doi.org / 10.1021 / jo00212a014 (1985)

[0351] All publications referenced herein are incorporated herein by reference. While exemplary embodiments of the present invention have been disclosed in detail herein with reference to the accompanying drawings, it should be understood that the present invention is not limited to these exact embodiments and that various modifications and changes can be made by those skilled in the art without departing from the scope of the invention as defined by the accompanying claims and equivalents.

Claims

1. Compound of formula (I): 【Chemistry 1】 Or a method for preparing a metallo-dera, salt, and solvate thereof, comprising the dicarboxylic acid compound of the following formula: 【Chemistry 2】 Or its salts and solvates, the following dialdehyde compounds: 【Transformation 3】 Alternatively, adding it to its salt and solvate and reacting it yields the following porphyrin diester compound: 【Chemistry 4】 or the step of forming a metallo-derivative, salt, and solvate thereof. [wherein, R １ , R ２ , R ３ , R ４ , R ５ , and R ６ are each independently H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', C １－２ haloalkyl, C １－２ alkanamine, and C(O)CH ３ ; R ７ and R ８ are independently selected from C １－６ alkyl; and R' and R'' are each independently selected from H and C １－６ alkyl] comprising said method.

2. The method according to claim 1, further comprising the step of hydrolyzing a porphyrin diester compound or its metallo derivative, salt, and solvate to obtain a dicarboxylic acid of formula (I), or its metallo derivative, salt, and solvate.

3. Preparation of dicarboxylic acid compounds, pyrrole A: 【Transformation 5】 Or its salts and solvates are pyrrole B: 【Transformation 6】 Alternatively, by reacting with its salt and solvate, the following diester compound is obtained: 【Transformation 7】 or a step of forming a salt and solvate thereof [wherein R ９ and R １０ The method according to claim 1 or 2, further comprising [ independently selected from benzyl, para-methoxybenzyl, and 2,4-dimethoxybenzyl].

4. The diester compound of the following formula: 【Transformation 8】 Alternatively, by subjecting its salt and solvate to a deprotection reaction, the following dicarboxylic acid compound is obtained: 【Chemistry 9】 The method according to claim 3, further comprising the step of forming a salt and solvate thereof.

5. The method according to any one of claims 1 to 4, further comprising the step of converting a compound of formula (I) or its metallo-derivative, salt, and solvate into a metallo-derivative that is a tin compound, wherein the tin compound is a tin mesylate compound, tin oxalate compound, tin phosphate compound, or tin tartrate compound of porphyrindicarboxylic acid of formula (I), or its salts and solvates.

6. The process further comprises the step of converting a compound of formula (I) or its metallo-derivative, salt, and solvate into a metallo-derivative that is a tin compound, wherein the tin compound is 【Chemistry 10】 The method according to any one of claims 1 to 5, or a salt or solvate thereof.

7. Compounds of formula (I) or their metallo-derivatives, salts, and solvates, as tin(IV) phosphate salts of formula (II) or (III): 【Chemistry 11】 The method according to any one of claims 1 to 6, further comprising the step of converting to a salt or solvate thereof.

8. A compound of formula (I) or its metallo derivatives, salts, and solvates, wherein the compound is 【Chemistry 12】 or its salts and solvates [wherein R １ , R ２ , R ３ , R ４ , R ５ , and R ６ These are independently H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', and C. １－２ Haloalkyl, C １－２ Alkanamines and C(O)CH ３ Selected from, R' and R'' are independently H and C １－６ The compound or its metallo-derivatives, salts, and solvates, which are selected from alkyl groups.

9. Compounds of formula (II) or (III): 【Chemistry 13】 or its salts and solvates [wherein R １ , R ２ , R ３ , R ４ , R ５ , and R ６ These are independently H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R'', and C. １－２ Haloalkyl, C １－２ Alkanamines and C(O)CH ３ Selected from, R' and R'' are independently H and C １－６ The compound according to claim 8, which is selected from alkyl groups.

10. Formula (VII) or (VIII): 【Chemistry 14】 Or it may be represented by its salts and solvates, R １ , R ３ , R ５ , and R ６ is methyl, R ２ and R ４ A compound of formula (II) or (III) according to claim 9, wherein is ethyl.

11. A compound of formula (I) according to any one of claims 8 to 10, or a salt and solvate thereof, for use in the treatment of proliferative and / or malignant diseases.

12. A compound of formula (I), or a salt and solvate thereof, for use in the treatment of a proliferative and / or malignant disease according to claim 11, wherein the proliferative and / or malignant disease is selected from breast cancer, lung cancer, brain tumors, and central nervous system cancers, carcinomas, gastrointestinal cancers, hormone-dependent cancers, leukemia, liver cancer, lymphoma, sarcoma and fibrosarcoma, skin cancer and melanoma, urinary tract cancer and genital cancer, and various other cancers.

13. A compound of formula (I), or a salt and solvate thereof, for use in the treatment of a proliferative and / or malignant disease according to claim 11 or 12, wherein the compound is administered simultaneously with or in succession to one or more immunotherapeutic agents or other anticancer agents.

14. A compound of formula (I) according to any one of claims 8 to 10, or a salt and solvate thereof, for use in the treatment or inhibition of metastasis.

15. A pharmaceutical composition comprising a compound of formula (I) according to any one of claims 8 to 10, or a salt and solvate thereof, and a pharmaceutically acceptable excipient, carrier, or diluent.

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