Porphyrin compound and medical application thereof

Abstract

The invention relates to a method for preparing porphyrin compounds of formula (I) or metal derivatives, salts and solvates thereof, in particular tin (IV) phosphate derivatives thereof: wherein each of R1, R2, R3, R4, R5 and R6 is independently selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR 'R '', C1-2 haloalkyl, C1-2 alkylamine and C (O) CH3. The invention also relates to the use of such porphyrins as medicaments, in particular as medicaments for the treatment of proliferative and / or malignant diseases, such as cancer, and for the treatment or inhibition of metastasis, and to pharmaceutical compositions containing such porphyrins.

Description

TECHNICAL FIELD

[0001] The present invention relates to methods of preparing porphyrins or metal derivatives, salts and solvates thereof, in particular tin mesoporphyrins, and to compounds prepared by such methods. The present invention also relates to the use of such porphyrins or metal derivatives, salts and solvates thereof as medicaments, in particular as medicaments for the treatment of proliferative and / or malignant diseases, such as cancer, and to pharmaceutical compositions containing such porphyrins. BACKGROUND

[0002] Cancer is a leading cause of death worldwide. There is still an urgent need for new cancer treatment methods to combat this disease.

[0003] One promising treatment approach involves immunotherapy targeting specific proteins. Preclinical data suggest that heme oxygenase-1 (HO-1) represents an important immunotherapeutic target in cancer (5).

[0004] The HO enzyme family is responsible for the breakdown of heme into the biologically active products biliverdin, ferrous (Fe 2+ ) and carbon monoxide (CO) (6). Carbon monoxide has been shown to have immunosuppressive effects (7-9), suggesting that it can play a role in blocking the immune response against tumors. Using an aggressive spontaneous breast cancer model (MMTV-PyMT), the present inventors have shown that the clinically relevant small molecule HO-1 inhibitor tin mesoporphyrin (SnMP) in combination with standard-of-care chemotherapy reduces immunosuppression and allows CD8+ T cells, immune cells that can specifically target cancer, to control tumor growth (5, 10).

[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. KCI-HO-1i is a tin phosphate salt of mesoporphyrin IX, the synthesis of which is shown in Scheme 2 below, and KCI-HO-1i is an HO inhibitor as shown in the Examples below. SnMP and KCI-HO-1i cross-react with HO-1 / 2.

[0006] Conventional SnMP synthesis uses chlorinated heme (typically from a bovine), which raises certification issues in certain markets and can create problems due to ethical, material procurement and health concerns.

[0007] SnMP as a chloride salt has been described in the literature as an inhibitor of HO-1 and HO-2. SnMP has shown tumour inhibitory effects in combination with existing chemotherapeutic or immunotherapeutic agents. The inventors' research has shown that SnMP in combination with standard of care chemotherapy enables NK cells to play a role in controlling tumour growth.

[0008] Infant hyperbilirubinaemia (also known as infant jaundice or neonatal hyperbilirubinaemia) occurs in newborns when the liver is unable to conjugate bilirubin and so can excrete bilirubin at a rate commensurate with its formation. Bilirubin arises from the release of haeme, which is part of the physiological transition from foetal to adult haemoglobin at birth. Stannsoporfin (tin (IV) dichloride mesoporphyrin IX) has been shown to have therapeutic value in treating hyperbilirubinaemia.

[0009]

[0010] Scheme 1 : mesoporphyrin IX.

[0011] The synthesis and use of SnMP has been described.

[0012] WO-A-2013 / 083659 discloses cancer therapies involving combinations of HO-1 inhibitors (such as SnMP) and immunotherapeutic agents.

[0013] JP-A-2013 / 232129 discloses the synthesis of stannsoporfin (tin (IV) dichloride mesoporphyrin IX) in high purity and quantity using chlorohaematin as a starting material, and the resulting composition.

[0014] US-B-10,533,024 discloses a method of synthesising metallo mesoporphyrin compounds by chlorohaematin metal transfer and subsequent tin protoporphyrin IX hydrogenation to form metallo mesoporphyrin.

[0015] US-B-8,530,458 discloses a method of synthesising stannsoporfin (tin (IV) dichloride mesoporphyrin IX) in high purity and quantity, and the composition thus prepared and its use for the treatment of infant hyperbilirubinaemia.

[0016] However, there is a need for improved methods of preparing mesoporphyrin and its derivatives, particularly methods for synthesis that do not rely on animal products (such as chlorohaematin). There is also a need for improved therapeutic forms of mesoporphyrin and its derivatives, including novel salt forms.

[0017] The inventors have found that certain tin salts of mesoporphyrin derivatives can provide improved treatments of proliferative and / or malignant diseases; and inhibition or reduction of metastasis.

[0018] The present invention seeks to address these needs and overcome problems associated with the prior art. SUMMARY

[0019] Accordingly, the present invention provides in a first aspect a process for the preparation of a compound of formula (I) or a metal derivative, salt and solvate thereof:

[0020]

[0021] wherein the process comprises the step of adding a dicarboxylic acid compound of formula:

[0022]

[0023] or a salt and solvate thereof to a dialdehyde compound of formula:

[0024]

[0025] or a salt and solvate thereof, and reacting to form a porphyrin diester compound of formula:

[0026]

[0027] or a metal derivative, salt and solvate thereof;

[0028] wherein each of R1, R2, R3, R4, R5and R6is independently selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR’R”, C 1-2 haloalkyl, C 1-2 alkylamine and C(O)CH3; and R7and R8are independently selected from C 1-6 alkyl; and R’ and R” are each independently selected from H and C 1-6 alkyl.

[0029] In another aspect, there is provided a compound of formula (I) or a metal derivative, salt and solvate thereof, wherein the compound is:

[0030] or a salt and solvate thereof.

[0031] More suitably, there is provided a compound of formula (II) or (III) of formula:

[0032] or a salt and solvate thereof.

[0033] More suitably, for a compound of formula (II) or (III), R1, R3, R5and R6are methyl; and R2and R4are ethyl, to give a compound of formula (VI) or (VII):

[0034]

[0035] Or its salts and solvates.

[0036] On the other hand, compounds of formula (II), (III), (IV), (V), or (VI) or their salts and solvates are provided for use as pharmaceuticals.

[0037] On the other hand, the use of compounds of formula (II), or (III), or (IV), or (V), or (VI), or salts thereof and solvates thereof, in the treatment of proliferative and / or malignant diseases is provided.

[0038] On the other hand, the use of compounds of formula (II), or (III), or (IV), or (V), or (VI), or their salts and solvates, in therapeutic applications is provided.

[0039] On the other hand, a pharmaceutical composition is provided comprising a compound of formula (II), or (III), or (IV), or (V), or (VI) or a salt thereof and a solvate thereof, and a pharmaceutically acceptable excipient, carrier or diluent.

[0040] On the other hand, the use of compounds of formula (II), or (III), or (IV), or (V), or (VI), or salts thereof and solvates thereof, in the preparation of medicaments for the treatment of proliferative and / or malignant diseases is provided.

[0041] On the other hand, a method for treating a patient suffering from proliferative and / or malignant diseases is provided, 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 thereof and a solvate thereof.

[0042] On the other hand, compounds of formula (II), or (III), or (IV), or (V), or (VI) or their salts and solvates are provided, wherein the compound is applied alone or in combination with one or more immunotherapeutic agents or other anticancer agents for the treatment of proliferative and / or malignant diseases, either simultaneously or sequentially.

[0043] On the other hand, methods for preparing compounds of formula (I) or their metal derivatives, salts, and solvates are provided:

[0044]

[0045] Each of R1, R2, R3, R4, R5, and R6 is independently selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", and C. 1-2 Haloalkyl, C 1-2alkyl, the method comprising the steps of: 1-6 alkyl, the method comprising the steps of:

[0046] (a) reacting a pyrrole A:

[0047] or salts and solvates thereof,

[0048] with a pyrrole B:

[0049] or salts and solvates thereof,

[0050] wherein R9and R 10 are independently selected from benzyl, p-methoxybenzyl and 2,4-dimethoxybenzyl;

[0051] to form a diester compound of the formula:

[0052] or salts and solvates thereof;

[0053] (b) subjecting the diester compound or salts and solvates thereof to a deprotection reaction to form a dicarboxylic acid compound of the formula: or salts and solvates thereof,

[0054] (c) adding the dicarboxylic acid compound or salts and solvates thereof to a dialdehyde compound of the formula:

[0055] or salts and solvates thereof, and reacting to form a porphyrin diester compound of the formula:

[0056]

[0057] or salts and solvates thereof, wherein R7and R8are each independently selected from C 1-6 alkyl;

[0058] (d) subjecting the porphyrin diester compound or salts and solvates thereof to hydrolysis to provide a porphyrin dicarboxylic acid of the formula (I):

[0059] or salts and solvates thereof; and

[0060] (e) converting the porphyrin dicarboxylic acid of the formula (I) or salts and solvates thereof to a metal derivative of the porphyrin dicarboxylic acid of the formula (I) or salts and solvates thereof; suitably, converting the porphyrin dicarboxylic acid of the formula (I) or salts and solvates thereof to a tin compound of the porphyrin dicarboxylic acid of the formula (I) or salts and solvates thereof.

[0061] In another aspect, the compounds of formula (II), or (III), or (IV), or (V), or (VI), or salts and solvates thereof, or mixtures thereof, can be administered alone or in combination with other treatments either simultaneously or sequentially, depending upon the condition to be treated.

[0062] In another aspect, there is provided the use of a compound of formula (II), or (III), or (IV), or (V), or (VI), or salts and solvates thereof, in the treatment or inhibition of metastasis.

[0063] Definitions

[0064] The following abbreviations are used throughout the specification: Ac acetyl -C(O)CH3; Et ethyl; Et3N triethylamine; 5-FU fluorouracil; KCl-HO-1 i tin phosphonate salt of mesoporphyrin IX; Me methyl; MeOH methanol; Ph (phenyl); p-TSA / PTSA p-toluenesulfonic acid; TBAC tetrabutylammonium chloride; TEOF triethyl orthoformate; TFA trifluoroacetic acid; THF tetrahydrofuran; and Ts (p-toluenesulfonyl salt) p-toluenesulfonyl group [-SO2-C6H4-CH3]; i.p. intraperitoneal; SD standard deviation; and SEM standard error of the mean.

[0065] Adoptive cell therapy is an immunotherapy that uses the subject's own immune cells (or donor immune cells) to treat a disease (e.g., cancer or viral infection). In adoptive cell therapy, T cells are isolated based on their ability to expand in response to a tumor, or are genetically modified to target certain molecules on cells, such as antigens on cancer cells. The tumor-reactive T cells are then expanded and infused back into the subject.

[0066] "TIL therapy" is a form of adoptive cell therapy that involves tumor infiltrating lymphocytes. In TIL therapy, tumor infiltrating lymphocytes that have penetrated a tumor are collected from a tumor biopsy taken from a subject and expanded in vitro for reinfusion into the patient. Tumor infiltrating lymphocytes actively participate in tumor destruction. In one method, after the biopsy is excised, DNA isolated from the tumor is sequenced to identify mutations found in the cancer that are recognized as neoantigens. In this method, the mutated neoantigens are inserted into autologous dendritic cells that are co-cultured with the tumor infiltrating lymphocytes. The tumor infiltrating lymphocytes for neoantigen recognition are then assayed. Those tumor infiltrating lymphocytes that recognize the neoantigens are then selected, expanded, and transferred for infusion back into the subject. In other methods, T cells are expanded in number due to their ability to recognize the tumor biopsy from which they were isolated, and are infused back into the patient.

[0067] Another approach to adoptive cell therapy is called TCR therapy, which involves engineering the T cells of the subject or donor to express a specific T cell receptor ("TCR"). The T cell receptor is a heterodimer composed of two subunits (TCRa and TCRp). Each subunit contains a constant region that anchors the receptor to the cell membrane and a hypervariable region that plays a role in antigen recognition. The TCR can recognize tumor-specific proteins / peptides presented on the outside of the cell. In TCR therapy, T cells are harvested from the blood of the subject or donor. The T cells are genetically modified in the lab to express a new T cell receptor. The T cells are expanded in number and infused back into the subject. The T cells with the new T cell receptor can target the patient's cancer.

[0068] Another adoptive cell therapy is chimeric antigen receptor ("CAR") T cell therapy ("CAR-T therapy"). In CAR-T therapy, one or more parts of the T cell receptor are changed to an antigen-binding moiety, such as an antibody, antibody fragment, or receptor ligand. Cancer-associated antigens (tumor-associated antigens or "TAAs") are typically expressed by tumors. The antibody or antibody fragment targets the TAA. The T cells targeting the TAA can directly attack the cancer cells. In CAR-T therapy, T cells are harvested from the blood of the subject or donor and genetically modified to express a CAR. The T cells are expanded in number and infused back into the subject. The CAR-T modification specifically targets the T cells to the subject's cancer.

[0069] "Substituted" when used in conjunction with chemical substituent or moiety (e.g., an alkyl group), means that one or more hydrogen atoms of the substituent or moiety have been replaced by one or more non-hydrogen atoms or groups, provided that chemically stable compounds are produced.

[0070] "Independently" or "independently selected" is used in the context of a statement such as "R1, R2 are each independently H, methyl, ethyl,...," and means that each instance of the functional group, e.g., R1, is selected independently from the listed options, independent of any other instance of R1 or R2 in the compound. Thus, for example, a first instance of R1 in a compound can be selected from H; another instance of R1 in the compound can be selected from methyl; and a first instance of R2 in the compound can be selected from ethyl.

[0071] C 1-6 alkyl: refers to straight chain and branched chain saturated hydrocarbon groups having generally from 1 to 6 carbon atoms; suitably, refers to C 1-5 alkyl; more suitably, refers to C 1-4 alkyl; more suitably, refers to C 1-3alkyl; more suitably, methyl or ethyl. Examples of alkyl groups include methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, t-butyl, pent-1-yl, pent-2-yl, pent-3-yl, 3-methylbut-1-yl, 3-methylbut-2-yl, 2-methylbut-2-yl, 2,2,2-trimethyleth-1-yl, n-hexyl, n-heptyl, n-octyl, and the like.

[0072] C 1-2 alkyl. Thus, C 1-6 alkyl. Thus, C 1-2 alkyl. Thus, C

[0073] C 1-2 haloalkyl. Thus, C 1-2 haloalkyl. Thus, C 1-2 haloalkyl. Thus, C 1-2 each halo in haloalkyl is the same.

[0074] “Halo” can be used interchangeably with “halogen” and each halo is independently selected from fluorine, chlorine, bromine, and iodine. Suitably, each halo is independently fluorine or chlorine.

[0075] The term “or a metal derivative, salt, and solvate thereof” means that the porphyrin compound can alternatively contain none or one or more of a metal group, salt, and solvate. Thus, the compound can be a metal derivative, salt, or solvate or any combination of these options. A metal derivative is a compound in which a metal is located at the center of the porphyrin ring. The most interesting metal group is tin. For example, a compound of formula (II) or (III) as described herein is a tin metal derivative of a compound of formula (I) that also contains a phosphate salt.

[0076] The term "oncolytic virus" is used according to its plain ordinary meaning and refers to a virus that preferentially infects and kills cancer cells. When infected cancer cells are destroyed by oncolysis (direct lysis of the cell), they release new infectious viral particles or virions to help destroy the remaining tumor. Oncolytic viruses are thought to cause not only direct destruction of tumor cells, but also to stimulate the host anti-tumor immune system response. Suitable oncolytic viruses are adenoviruses, reoviruses, measles viruses, herpes simplex viruses, Newcastle disease viruses, vaccinia viruses, or senecaviruses.

[0077] The term "or a pharmaceutically acceptable salt, solvate, tautomer, stereoisomer, or mixture thereof" means that pharmaceutically acceptable salts, solvate, tautomer, stereoisomer forms of the depicted structure are also included. Mixtures thereof means that mixtures of these forms can exist, for example, a compound of the application can include both tautomeric forms and pharmaceutically acceptable salts. "Pharmaceutically acceptable" means those substances that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of subjects without excessive toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio, and effective for their intended purpose.

[0078] "Pharmaceutical composition" means a combination of one or more pharmaceutical substances and one or more excipients.

[0079] The term "or a salt and solvate thereof" means that the compound can alternatively comprise no salt and solvate or one or more of a salt and solvate. Thus, the compound can be a salt, the compound can be a solvate, or the compound can be both a salt and a solvate. Suitably, the broader term "or a metal derivative, salt and solvate thereof" can be limited to "or a salt and solvate thereof". Where the compound comprises a salt, there can be one or more salts, for example, there can be a case where only one of the carboxylic acid groups is a salt, or other cases where both carboxylic acid groups are salts. Suitably, any salt is a pharmaceutically acceptable salt.

[0080] As used herein, "solvate" means a complex of variable stoichiometry formed by a solute and a solvent. For crystalline compounds, a pharmaceutically acceptable solvate can be formed, in which the solvent molecules are incorporated in the crystalline lattice during the process of crystallization. The incorporated solvent molecules can be water molecules or non- water molecules such as, but not limited to, ethanol, isopropanol, dimethylsulfoxide, acetic acid, ethanolamine, and ethyl acetate molecules.

[0081] "Tautomer" means two or more isomers of a compound that exist in an equilibrium and are readily interchangeable by the shift of a proton and an electron pair between two atoms.

[0082] As used herein, the term "subject" refers to a human or a non-human mammal. Examples of non-human mammals include livestock, such as sheep, horses, cattle, pigs, goats, rabbits, and deer; and companion animals, such as cats, dogs, rodents, and horses. Suitably, the subject is a human.

[0083] A "therapeutically effective amount" of a drug refers to the amount of the drug or composition that is effective to treat a subject and thus produce the desired therapeutic, ameliorative, inhibitory or preventative effect. The therapeutically effective amount can depend on the weight and age of the subject, and the route of administration, among other things.

[0084] "Treating" refers to reversing, alleviating, inhibiting the progress of, or preventing the disorder, disease, or condition to which such term applies, or to reversing, alleviating, inhibiting the progress of, or preventing one or more symptoms of such disorder, disease, or condition.

[0085] "Treatment" refers to the act of "treating," as defined above.

[0086] As used herein, the term "comprising" means "including at least" and is intended to be

[0087] The term "consisting essentially of shall limit the scope of a claim to the specified materials or steps "plus" those that do not materially affect the basic and novel characteristic(s) of the claimed application. When the phrase "consisting essentially of appears in the clause of a claim, it shall be construed in accordance with 35 U.S.C. § 112, paragraph six.

[0088] The term“consisting of’ excludes any element, step, or ingredient not specified in the claim; defines the claim as encompassing only those elements recited in the claim, to the exclusion of additional materials otherwise commonly utilized in the practice of the invention. When the phrase“consisting of’ appears in a clause of the body of the claim, it limits the elements set forth in that clause to the exclusion of additional elements not specified in that clause. It is to be understood that while various embodiments have been presented in the specification by way of example, related embodiments also exist.

[0089] Deprotection reaction

[0090] Suitably, the method further comprises the step of subjecting a di-ester compound of the formula:

[0091]

[0092] or salts and solvates thereof to a deprotection reaction to form a dicarboxylic acid compound of the formula:

[0093]

[0094] or salts and solvates thereof.

[0095] More suitably, the method comprises the step of subjecting a di-ester compound of the formula:

[0096] or salts and solvates thereof to a deprotection reaction to form a dicarboxylic acid compound of the formula:

[0097] or salts and solvates thereof.

[0098] Suitably, the di-ester compound or salts and solvates thereof is subjected to a deprotection reaction to form the dicarboxylic acid compound using: (i) tin and concentrated HC1; (ii) iron formate and ammonium formate; (iii) CrO3 and acetic acid; (iv) N-bromosuccinimide (NBS) and ozone; or (v) a hydrogenation reaction.

[0099] Suitably, the deprotection reaction is carried out by refluxing the di-ester compound or salts and solvates thereof using (i) tin and concentrated HC1.

[0100] Suitably, the deprotection reaction is carried out using (ii) iron formate and ammonium formate in ethanol and water.

[0101] Suitably, the deprotection reaction is carried out using (iii) CrO3 and acetic acid.

[0102] Suitably, the deprotection reaction is a (v) hydrogenation reaction. Suitably, the hydrogenation reaction is carried out using a Pd / C catalyst.

[0103] Suitably, the deprotection reaction is a (v) hydrogenation reaction. Suitably, the hydrogenation reaction is carried out using a Pd / C catalyst.

[0104] Suitably, the hydrogenation reaction is carried out in the presence of an organic base.

[0105] Suitably, the hydrogenation is carried out in the presence of triethylamine.

[0106] Diester formation

[0107] Suitably, the method comprises the step of reacting a pyrrole A:

[0108] or salts and solvates thereof,

[0109] with a pyrrole B:

[0110] or salts and solvates thereof,

[0111] to form a diester compound of the formula:

[0112] or salts and solvates thereof.

[0113] Suitably, the pyrrole A is:

[0114] or salts and solvates thereof.

[0115] More suitably, R1is methyl and the method comprises the step of reacting a pyrrole A:

[0116] or salts and solvates thereof, with a pyrrole B:

[0117] or salts and solvates thereof,

[0118] to form a diester compound of the formula:

[0119]

[0120] Suitably, pyrrole A is reacted with pyrrole B in the presence of a protic acid or a montmorillonite clay. Suitably, pyrrole A is reacted with pyrrole B in the presence of a protic acid. Suitably, pyrrole A is reacted with pyrrole B in the presence of a montmorillonite clay. Suitably, the protic acid is a mineral acid, an organic sulfonic acid or trifluoroacetic acid. Suitably, the mineral acid is boric acid, hydrobromic acid, hydrochloric acid, hydrofluoric acid, hydroiodic acid, nitric acid, perchloric acid, phosphoric acid or sulfuric acid. Suitably, the organic sulfonic acid is benzenesulfonic acid, camphorsulfonic acid, ethanesulfonic acid, methanesulfonic acid or p-toluenesulfonic acid. More suitably, the protic acid is p-toluenesulfonic acid. Suitably, the montmorillonite clay is K10 montmorillonite clay.

[0121] Suitably, pyrrole A is reacted with pyrrole B in methanol.

[0122] Porphyrin diester formation

[0123] Suitably, the method comprises the step of adding a dicarboxylic acid compound of the formula:

[0124] or salts and solvates thereof, to a dialdehyde compound of the formula: or salts and solvates thereof, and reacting to form a porphyrin diester compound of the formula:

[0125] or metal derivatives, salts and solvates thereof.

[0126] More suitably, a dicarboxylic acid compound of the formula:

[0127] or salts and solvates thereof, to a dialdehyde compound of the formula: or salts and solvates thereof, and reacting to form a porphyrin diester compound of the formula:

[0128] or metal derivatives, salts and solvates thereof.

[0129] Suitably, (i) a dicarboxylic acid compound or salt and solvate thereof is added to a dialdehyde compound or salt and solvate thereof in the presence of a protic acid or a montmorillonite clay. Suitably, the addition is in the presence of a protic acid. Suitably, the addition is in the presence of a montmorillonite clay. Suitably, the protic acid is a mineral acid, an organic sulfonic acid or trifluoroacetic acid. Suitably, the mineral acid is boric acid, hydrobromic acid, hydrochloric acid, hydrofluoric acid, hydroiodic acid, nitric acid, perchloric acid, phosphoric acid or sulfuric acid. Suitably, the organic sulfonic acid is benzenesulfonic acid, camphorsulfonic acid, ethanesulfonic acid, methanesulfonic acid or p-toluenesulfonic acid. More suitably, the protic acid is p-toluenesulfonic acid. Suitably, the montmorillonite clay is K10 montmorillonite clay.

[0130] Suitably, (i) the dicarboxylic acid compound or salt and solvate thereof is added to the dialdehyde compound or salt and solvate thereof using a chloroform and methanol solvent system. Suitably, (i) the chloroform is the major component (i.e. greater than 50% by volume) of the chloroform and methanol solvent system. Suitably, (i) the chloroform is present in a ratio of chloroform:methanol of at least 2:1; suitably, in a ratio of at least 3:1; suitably, in a ratio of at least 4:1. Suitably, (i) the chloroform:methanol is present in a ratio of about 5:1.

[0131] Suitably, after the addition of the dicarboxylic acid compound or salt and solvate thereof and the dialdehyde compound or salt and solvate thereof, (ii) the intermediate is treated using zinc acetate and oxygen; or using zinc hexafluoroacetylacetonate dihydrate. More suitably, (ii) the intermediate is treated using zinc acetate and oxygen.

[0132] Suitably, after the treatment using zinc acetate and oxygen or using zinc hexafluoroacetylacetonate dihydrate, (iii) the further intermediate is treated using trifluoroacetic acid.

[0133] Hydrolysis of porphyrin diester

[0134] Suitably, the method further comprises the step of hydrolysing a porphyrin diester compound of the formula:

[0135]

[0136] or a metal derivative, salt and solvate thereof, to provide the dicarboxylic acid of the formula (I): or a metal derivative, salt and solvate thereof.

[0137] More suitably, the method further comprises the step of hydrolysing a porphyrin diester compound of the formula: or a metal derivative, salt and solvate thereof, to provide the dicarboxylic acid of the formula (I):

[0138] or a metal derivative, salt and solvate thereof.

[0139] Suitably, the hydrolysis step is a base hydrolysis step. Suitably, the base hydrolysis step is carried out using sodium hydroxide or lithium hydroxide.

[0140] Suitably, the hydrolysis step is carried out in the presence of tributylammonium chloride (TBAC).

[0141] Suitably, the hydrolysis step is carried out in a dichloromethane and methanol solvent system.

[0142] Suitably, the hydrolysis step is carried out in methanol.

[0143] In some aspects, hydrolysis is performed using lithium hydroxide in ethanol.

[0144] Conversion to metal derivative

[0145] Suitably, the method further comprises the step of converting the compound of formula (I) to a metal derivative of a porphyrin dicarboxylic acid of formula (I), or salts and solvates thereof.

[0146] Suitably, the metal derivative is a tin compound of a porphyrin dicarboxylic acid of formula (I), or salts and solvates thereof. More suitably, the metal derivative is a tin methanesulfonate compound, a tin oxalate compound, a tin phosphate compound, or a tin tartrate compound of a porphyrin dicarboxylic acid of formula (I), or salts and solvates thereof.

[0147] Suitably, the method further comprises the step of converting the compound of formula (I), or a metal derivative, salt and solvate thereof, to a tin (IV) compound, or a salt and solvate thereof. Suitably, the compound of formula (I), or a salt and solvate thereof, is contacted with a tin source to produce a tin compound of a porphyrin dicarboxylic acid of formula (I), or a salt and solvate thereof.

[0148] Suitably, the method further comprises the step of converting the compound of formula (I), or a metal derivative, salt and solvate thereof, to a metal derivative which is a tin compound, wherein the tin compound is a tin acetate compound, a tin methanesulfonate compound, a tin oxalate compound, a tin phosphate compound, or a tin tartrate compound of a porphyrin dicarboxylic acid of formula (I), or a salt and solvate thereof.

[0149] Suitably, the method further comprises the step of converting the compound of formula (I), or a metal derivative, salt and solvate thereof, to a metal derivative which is a tin compound, wherein the tin compound is a tin methanesulfonate compound, a tin oxalate compound, a tin phosphate compound, or a tin tartrate compound of a porphyrin dicarboxylic acid of formula (I), or a salt and solvate thereof.

[0150] More suitably, the method further comprises the step of converting the compound of formula (I), or a metal derivative, salt and solvate thereof, to a tin (IV) compound of formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof.

[0151] More suitably, the method further comprises the step of converting the compound of formula (I), or a metal derivative, salt and solvate thereof, to a metal derivative which is a tin compound, wherein the tin compound is:

[0152]

[0153]

[0154] or salts and solvates thereof.

[0155] Suitably, the compound of formula (I) or a metal derivative, salt and solvate thereof is converted to a tin(IV) compound using SnO or tin(II) chloride.

[0156] Suitably, the reaction using SnO is carried out in the presence of HCO2H. Suitably, the reaction using SnO is carried out in the presence of CH3CO2H.

[0157] Suitably, the reaction using tin(II) chloride is carried out in the presence of acetic acid. Suitably, the compound of formula (I) or a salt and solvate thereof is refluxed in acetic acid using tin(II) chloride.

[0158] Suitably, after the reaction using SnO, the intermediate is treated using an acid. Suitably, after the reaction using SnO, the intermediate is treated using hydrochloric acid.

[0159] In other aspects, more suitably, the method further comprises the step of converting the compound of formula (I) or a metal derivative, salt and solvate thereof to a tin(IV) phosphate of formula (II) or (III):

[0160]

[0161] or salts and solvates thereof.

[0162] with a source of tin and a source of phosphate to produce

[0163] More suitably, the compound of formula (I) or a metal derivative, salt and solvate thereof is converted to a tin(IV) compound using tin(II) pyrophosphate. More suitably, the reaction using tin(II) pyrophosphate is carried out in the presence of acetic acid. More suitably, the compound of formula (I) or a metal derivative, salt and solvate thereof is refluxed in acetic acid using tin(II) pyrophosphate.

[0164] More suitably, the method further comprises the step of converting the compound of formula (I) or a metal derivative, salt and solvate thereof to a tin(IV) phosphate salt of formula (VII) or (VIII):

[0165]

[0166] or salts and solvates thereof.

[0167] Compound

[0168] Suitably, the compound of formula (I) or a metal derivative, salt and solvate thereof is a tin salt of the compound of formula (I), wherein the tin salt is a tin acetate salt, a tin methylsulfonate salt, a tin oxalate salt, a tin phosphate salt or a tin tartrate salt.

[0169] Suitably, the tin salt of the compound of formula (I) is a tin methanesulfonate salt, a tin oxalate salt, a tin phosphate salt or a tin tartrate salt. More suitably, the tin salt is a tin methanesulfonate salt, a tin oxalate salt or a tin phosphate salt; or alternatively, the tin salt is a tin oxalate salt, a tin phosphate salt or a tin tartrate salt; or alternatively, the tin salt is a tin methanesulfonate salt, a tin phosphate salt or a tin tartrate salt.

[0170] More suitably, the tin salt is a tin methanesulfonate salt or a tin phosphate salt; or alternatively, the tin salt is a tin oxalate salt or a tin phosphate salt or a tin tartrate salt.

[0171] a tin salt of the compound of formula (I), wherein the tin salt is a tin acetate salt, a tin methanesulfonate salt, a tin oxalate salt or a tin phosphate salt.

[0172] More suitably, for any of the above tin salts of the compound of formula (I), the substituents R1, R3, R5and R6are methyl groups; and R2and R4are ethyl groups.

[0173] Suitably, the compound of formula (I) or a metal derivative, salt and solvate thereof is a compound of formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof.

[0174] More suitably, the compound of formula (I) or a metal derivative, salt and solvate thereof is a compound of formula (II), or (III), or (IV), or (V), or (VI), wherein the substituents R1, R3, R5and R6are methyl groups; and R2and R4are ethyl groups. Such compounds have the following formula:

[0175]

[0176] or a salt and solvate thereof.

[0177] Synthesis of pyrrole A

[0178] Suitably, the method further comprises the step of: (i) reacting a compound of formula:

[0179] or a salt and solvate thereof, to produce an oxime of formula:

[0180] or a salt and solvate thereof.

[0181] More suitably, step (i) is reacting a compound of formula:

[0182]

[0183] or salts and solvates thereof, to produce an oxime of the formula:

[0184] or salts and solvates thereof.

[0185] Suitably, step (i) uses sodium nitrite and acetic acid to prepare the oxime.

[0186] Suitably, the method further comprises the step of: (ii) subjecting the oxime, or a salt and solvate thereof, to a cyclisation reaction with R2-CH2C(O)CH3to produce a pyrrole of the formula:

[0187] or salts and solvates thereof.

[0188] Suitably, R2is acetyl, such that R2-CH2C(O)CH3is acetylacetone, and the cyclisation reaction step (ii) of the oxime, or a salt and solvate thereof, with acetylacetone produces a pyrrole of the formula:

[0189] or salts and solvates thereof.

[0190] Suitably, the cyclisation reaction of step (ii) is performed in the presence of one or more of sodium acetate, acetic acid and zinc.

[0191] Suitably, the method further comprises the step of: (iii) subjecting the compound of the formula:

[0192]

[0193] or salts and solvates thereof, to a selective reduction to produce a pyrrole compound of the formula:

[0194]

[0195] or salts and solvates thereof.

[0196] Suitably, the selective reduction is a Wolf-Kishner reduction, a Clemmenson reduction, a catalytic hydrogenation or a reduction using TsNHNH2followed by NaBH4CN, such selective reductions being well known in the literature (11). Suitably, the Wolf-Kishner reduction is performed using NH2NH2and KOH. Suitably, the Clemmenson reduction is performed using Zn-Hg and HC1. Suitably, the catalytic hydrogenation is performed using Pt-C, H2under pressure.

[0197] Suitably, the method further comprises the step of: (iv) subjecting the pyrrole compound of the formula:

[0198]

[0199] or salts and solvates thereof, to add an acyloxy group to form a pyrrole A:

[0200]

[0201] or salts and solvates thereof.

[0202] Suitably, step (iv) is catalysed using lead tetraacetate.

[0203] More suitably, R1is methyl and step (iv) forms a pyrrole A:

[0204] or salts and solvates thereof.

[0205] Synthesis of pyrrole B

[0206] Suitably, the method further comprises the step of: (a) converting an amine of the formula:

[0207]

[0208] or salts and solvates thereof, to an isocyanide of the formula:

[0209]

[0210] or salts and solvates thereof. Suitably, step (a) is performed using KOH and benzyltrimethylammonium bromide.

[0211] Suitably, the method further comprises the step of: (b) performing a cyclisation reaction between an isocyanide of the formula:

[0212]

[0213] or salts and solvates thereof, and an alkyne of the formula:

[0214]

[0215] or salts and solvates thereof, to form a pyrrole compound of the formula:

[0216] or salts and solvates thereof.

[0217] Suitably, R4is C(O)CH3and the alkyne has the formula:

[0218] or salts and solvates thereof.

[0219] This type of cyclisation reaction has been discussed in the literature (12). Suitably, the cyclisation step (b) is carried out using a Cu catalyst. Suitably, the cyclisation step (b) is carried out using a Cu catalyst in the presence of triphenylphosphine (PPh3). Suitably, the Cu catalyst is Cu2O. Suitably, the Cu catalyst is present at 1 to 10 mol%; suitably, at 4 to 6 mol%; more suitably, at 5 mol%. Suitably, the triphenylphosphine is present at 10 to 30 mol%; suitably, at 15 to 25 mol%; more suitably, at 20 mol%. Suitably, R4is C(O)CH3, and the method further comprises the step of: (c) selective reduction of the pyrrole compound:

[0220]

[0221] or salts and solvates thereof, to form a pyrrole B:

[0222] or salts and solvates thereof.

[0223] More suitably, R3is methyl, and step (c) forms a pyrrole B:

[0224] or salts and solvates thereof.

[0225] Suitably, the selective reduction is a Wolff-Kishner reduction, Clemmensen reduction, catalytic hydrogenation or reduction using TsNHNH2followed by NaBH4CN, such selective reductions are well known in the literature (11). Suitably, the Wolff-Kishner reduction is carried out using NH2NH2and KOH. Suitably, the Clemmensen reduction is carried out using Zn-Hg and HC1. Suitably, the catalytic hydrogenation is carried out using Pt-C, H2under pressure.

[0226] Synthesis of dialdehyde

[0227] Suitably, the method further comprises the step of: (i) reacting a compound of the formula:

[0228]

[0229] or salts and solvates thereof, to produce an oxime of the formula:

[0230] or salts and solvates thereof.

[0231] Suitably, step (i) uses sodium nitrite and acetic acid to produce the oxime.

[0232] Suitably, the method further comprises the step of: (ii) reacting the oxime or salts and solvates thereof with a compound of the formula: carrying out a cyclisation reaction to prepare a pyrrole of the formula:

[0233] or salts and solvates thereof.

[0234] Suitably, the method further comprises the step of (ia) reacting a compound of the formula:

[0235]

[0236] or salts and solvates thereof, to prepare an oxime of the formula:

[0237] or salts and solvates thereof.

[0238] Suitably, the method further comprises the step of (iia) reacting the oxime, or a salt and solvate thereof, with a compound of the formula:

[0239] carrying out a cyclisation reaction to prepare a pyrrole of the formula:

[0240]

[0241] Suitably, the cyclisation reaction of step (iia) is carried out in the presence of one or more of sodium acetate, acetic acid and zinc.

[0242] Suitably, the method further comprises step (iii) wherein a pyrrole compound of the formula:

[0243] or salts and solvates thereof;

[0244] is coupled with a pyrrole compound of the formula:

[0245] or salts and solvates thereof;

[0246] to thereby prepare a pyrrole diester compound of the formula:

[0247] or salts and solvates thereof.

[0248] Suitably, the method further comprises the step of (i) reacting a compound of the formula:

[0249] or salts and solvates thereof, to prepare an oxime of the formula:

[0250] or salts and solvates thereof.

[0251] Suitably, the method further comprises the step of (ii) reacting the oxime, or a salt and solvate thereof, with a compound of the formula:

[0252] The cyclisation reaction is carried out to prepare a pyrrole of the formula:

[0253]

[0254] or salts and solvates thereof.

[0255] Suitably, step (i) or (ia) uses sodium nitrite and acetic acid to prepare the oxime.

[0256] Suitably, the cyclisation reaction of step (ii) or (iia) is carried out in the presence of one or more of sodium acetate, acetic acid and zinc. Suitably, the cyclisation reaction of step (ii) or (iia) is carried out in the presence of sodium acetate, acetic acid and zinc.

[0257] Suitably, step (iii) uses Br2as a reagent for preparing the pyrrole diester compound. Suitably, step (iii) is carried out in methanol.

[0258] Suitably, step (iii) couples two identical pyrrole compounds (i.e. R5= R6and R7= R8).

[0259] Suitably, the method further comprises step (iv) wherein a pyrrole diester compound of the formula:

[0260]

[0261] or salts and solvates thereof is subjected to selective hydrolysis to prepare a pyrrole diacid compound of the formula:

[0262] or salts and solvates thereof.

[0263] Suitably, the selective hydrolysis of step (iv) is carried out under hydrogenation conditions.

[0264] Suitably, the selective hydrolysis of step (iv) is carried out using a Pd / C catalyst.

[0265] Suitably, the method further comprises step (v) selective reduction of a pyrrole diacid compound of the formula:

[0266]

[0267] or salts and solvates thereof to a dialdehyde compound of the formula:

[0268]

[0269] or salts and solvates thereof.

[0270] R1

[0271] R1is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3. Suitably, R1is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, and C 1-2 alkylamine.

[0272] Suitably, in some aspects, R1is selected from methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3.

[0273] In alternative aspects, R1is H.

[0274] More suitably, R1is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, and NR'R". More suitably, R1is selected from methyl, ethyl, methoxy, and ethoxy.

[0275] More suitably, R1is methyl or ethyl. Most suitably, R1is methyl.

[0276] R2

[0277] R2is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3. Suitably, R2is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, and C 1-2 alkylamine.

[0278] Suitably, in some aspects, R2is selected from methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3.

[0279] In alternative aspects, R2is H.

[0280] More suitably, R2is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", and C(O)CH3. More suitably, R2is selected from methyl, ethyl, methoxy, ethoxy, and C(O)CH3.

[0281] More suitably, R2is methyl or ethyl. Most suitably, R2is ethyl.

[0282] R3

[0283] R3is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3. Suitably, R3is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, and C 1-2 alkylamine.

[0284] Suitably, in some aspects, R3is selected from methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3.

[0285] In alternative aspects, R3is H.

[0286] More suitably, R3is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, and NR'R". More suitably, R3is selected from methyl, ethyl, methoxy, and ethoxy.

[0287] More suitably, R3is methyl or ethyl. Most suitably, R3is methyl.

[0288] R4

[0289] R4is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3. Suitably, R4is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, and C 1-2 alkylamine.

[0290] Suitably, in some aspects, R4is selected from methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3.

[0291] In alternative aspects, R4is H.

[0292] More suitably, R4is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R" and C(O)CH3. More suitably, R4is selected from methyl, ethyl, methoxy, ethoxy, and C(O)CH3.

[0293] More suitably, R4is methyl or ethyl. Most suitably, R4is ethyl.

[0294] R5

[0295] R5is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3. Suitably, R5is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, and C 1-2 alkylamine.

[0296] Suitably, in some aspects, R5is selected from methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3.

[0297] In alternative aspects, R5is H.

[0298] More suitably, R5is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, and NR'R". More suitably, R5is selected from methyl, ethyl, methoxy, and ethoxy.

[0299] More suitably, R5is methyl or ethyl. Most suitably, R5is methyl.

[0300] R6

[0301] R6is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3. Suitably, R6is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, and C 1-2 alkylamine.

[0302] Suitably, in some aspects, R6is selected from methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine, and C(O)CH3.

[0303] In alternative aspects, R6is H.

[0304] More suitably, R6is selected from H, methyl, ethyl, halogen, methoxy, ethoxy, and NR'R". More suitably, R6is selected from methyl, ethyl, methoxy, and ethoxy.

[0305] More suitably, R6is methyl or ethyl. Most suitably, R6is methyl.

[0306] R1, R2, R3, R4, R5, and R6

[0307] Suitably, one, two or three of R1, R2, R3, R4, R5and R6are independently selected from H, halo, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine and C(O)CH3; the remainder of R1, R2, R3, R4, R5and R6are independently selected from methyl and ethyl.

[0308] Suitably, one, two or three of R1, R3, R5and R6are independently selected from H, ethyl, halo, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine and C(O)CH3; the remainder of R1, R3, R5and R6are methyl; and R2and R4are ethyl. More suitably, one of R1, R3, R5and R6is independently selected from H, ethyl, halo, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine and C(O)CH3; the remainder of R1, R3, R5and R6are methyl; and R2and R4are ethyl.

[0309] Additionally, suitably, optionally, one or two of R1, R3, R5and R6are independently selected from H, ethyl, halo, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine and C(O)CH3; the remainder of R1, R3, R5and R6are methyl; and one of R2and R4is selected from H, methyl, halo, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine and C(O)CH3; and the other of R2and R4is ethyl. Thus, when this option is not taken, R1, R3, R5and R6are methyl.

[0310] Additionally, suitably, optionally, one of R1, R3, R5and R6is independently selected from H, ethyl, halo, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine and C(O)CH3; the remainder of R1, R3, R5and R6are methyl; and R2and R4are independently selected from H, methyl, halo, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, C 1-2 alkylamine and C(O)CH3. Thus, when this option is not taken, R1, R3, R5and R6are methyl.

[0311] For any of the four preceding paragraphs, suitably, the relevant number of R1, R3, R5, and R6is independently selected from H, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, and C 1-2 alkylamine. More suitably, the relevant number of R1, R3, R5, and R6is independently selected from H, ethyl, halogen, methoxy, ethoxy, and NR'R". More suitably, the relevant number of R1, R3, R5, and R6is independently selected from ethyl, methoxy, and ethoxy.

[0312] For any of the four preceding paragraphs, suitably, the relevant number of R1, R3, R5, and R6is independently selected from H, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 haloalkyl, and C 1-2 alkylamine. More suitably, the relevant number of R1, R3, R5, and R6is independently selected from H, ethyl, halogen, methoxy, ethoxy, and NR'R". More suitably, the relevant number of R1, R3, R5, and R6is independently selected from ethyl, methoxy, and ethoxy.

[0313] More suitably, R1, R3, R5, and R6are methyl; and R2and R4are ethyl.

[0314] R7

[0315] R7is C 1-6 alkyl. Suitably, R7is selected from C 1-5 alkyl. More suitably, R7is selected from C 1-4 alkyl.

[0316] Suitably, R7is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, and t-butyl. Suitably, R7is selected from methyl, ethyl, n-propyl, and i-propyl. More suitably, R7is selected from methyl and ethyl. Most suitably, R7is methyl.

[0317] R8

[0318] R8is C 1-6 alkyl. Suitably, R8is selected from C 1-5 alkyl. More suitably, R8is selected from C 1-4 alkyl.

[0319] Suitably, R8is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl, and t-butyl. Suitably, R8is selected from methyl, ethyl, n-propyl, and i-propyl. More suitably, R8is selected from methyl and ethyl. Most suitably, R8is methyl.

[0320] Suitably, R7and R8are the same.

[0321] R9

[0322] R9is independently selected from benzyl, p-methoxybenzyl and 2,4-dimethoxybenzyl. Suitably, R9is selected from benzyl and p-methoxybenzyl. More suitably, R9is benzyl.

[0323] R 10

[0324] R 10 is independently selected from benzyl, p-methoxybenzyl and 2,4-dimethoxybenzyl. Suitably, R 10 is selected from benzyl and p-methoxybenzyl. More suitably, R 10 is benzyl.

[0325] Suitably, R9and R 10 are the same.

[0326] NR'R ”

[0327] For 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 aspects, NR’R” is NH2.

[0328] Suitably, in other aspects, 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 NHCH3or NHCH2CH3. More suitably, NR’R” is NHCH3.

[0329] Proliferative and / or malignant disease

[0330] The use of a compound of formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof, in the treatment of a proliferative and / or malignant disease is described.

[0331] Suitably, the proliferative and / or malignant disease can be a metastatic or non-metastatic cancer. The cancer can be familial or sporadic. Suitably, the proliferative and / or malignant disease that can be treated can include, for example, benign or in situ lesions and malignant solid tumours as well as benign and malignant non-solid tumours.

[0332] For example, the proliferative and / or malignant disease can include a solid tumor, such as a carcinoma or sarcoma. Carcinomas include malignant tumors derived from epithelial cells that infiltrate (e.g., invade) surrounding tissues and give rise to metastases. Adenocarcinomas are carcinomas derived from glandular tissue or tissue that forms recognizable glandular structures.

[0333] The present application finds use in the treatment of proliferative and / or malignant diseases.

[0334] In certain aspects, there is provided a method for treating a proliferative and / or malignant disease, the method comprising 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.

[0335] Suitably, the proliferative and / or malignant disease is selected from breast cancer, lung cancer, brain and central nervous system cancer, carcinoma, gastrointestinal cancer, hormonal cancer, leukemia, liver cancer, lymphoma, sarcoma and fibrosarcoma, skin cancer and melanoma, urinary tract cancer and reproductive cancer, and various other cancers.

[0336] Suitably, the proliferative and / or malignant disease is selected from brain and central nervous system cancer, carcinoma, gastrointestinal cancer, hormonal cancer, leukemia, liver cancer, lymphoma, sarcoma and fibrosarcoma, skin cancer and melanoma, urinary tract cancer and reproductive cancer, and various other cancers.

[0337] Suitably, the brain and central nervous system (CNS) cancers and tumors that can be treated include astrocytic tumors (including cerebellar and cerebral), brain stem gliomas, brain tumors, malignant gliomas, ependymal tumors, glioblastomas, medulloblastomas, supratentorial primitive neuroectodermal tumors, visual pathway and hypothalamic glioma, primary central nervous system lymphoma, ependymal tumors, brain stem gliomas, visual pathway and hypothalamic glioma, extracranial germ cell tumors, medulloblastomas, myelodysplastic syndrome, oligodendroglioma, myelodysplasia / myeloproliferative disease, myelogenous leukemia, myeloid leukemia, multiple myeloma, myeloproliferative disease, neuroblastoma, plasma cell neoplasm / multiple myeloma, central nervous system lymphoma, intrinsic brain tumors, astrocytic brain tumors, gliomas, and / or metastatic tumor cell invasion in the central nervous system.

[0338] Treatable cancers include adrenocortical carcinoma, acinar carcinoma, acinic cell carcinoma, adenocystic carcinoma, adenoid cystic carcinoma, adenoid squamous cell carcinoma, adenomatosum carcinoma, adenosquamous carcinoma, adnexal carcinoma, cancer of the adrenal cortex, adrenocortical carcinoma, aldosterone-producing carcinoma, aldosterone-secreting carcinoma, alveolar carcinoma, alveolar cell carcinoma, ameloblastic carcinoma, ampullary carcinoma, and anaplastic cancer of the thyroid. (gland), apocrine carcinoma, basal cell carcinoma, alveolar carcinoma, comedo basal cell carcinoma, cystic basal cell carcinoma, morphea-like basal cell carcinoma, multicentric basal cell carcinoma, nodulo-ulcerative basal cell carcinoma, pigmented basal cell carcinoma, sclerosing basal cell carcinoma, superficial basal cell carcinoma, basaloid carcinoma, basosquamous cell carcinoma, bile duct carcinoma, extrahepatic bile duct carcinoma, intrahepatic bile duct carcinoma. Bronchioloalveolar carcinoma, bronchiolar carcinoma, bronchioloalveolar carcinoma, bronchial alveolar carcinoma, bronchoalveolar cell carcinomacancer, bronchogenic cancer, cerebriform cancer, cholangiocellular cancer, chorionic cancer, choroids plexus cancer, clear cell cancer, cloacogenic anal cancer, colloid cancer, comedo cancer, corpus cancer, cancer of corpus uteri, Cortisol-producing cancer, cribriform cancer, cylindrical cancer, cylindrical cell cancer, duct cancer, ductal cancer, ductal cancer of the prostate, ductal cancer in situ (DCIS), eccrine cancer, embryonal cancer, cancer en cuirasse, endometrial cancer, cancer of endometrium, endometroid cancer, epidermoid cancer, cancer ex mixed tumour, cancer ex pleomorphic adenoma, exophytic cancer, fibrolamellar cancer, cancer fibrosum, follicular cancer of thyroid gland, gastric cancer, gelatinform cancer, gelatinous cancer, giant cell cancer, giant cell cancer of thyroid gland, cancer gigantocellular, glandular cancer, granulose cell cancer, hepatocellular cancer, Hurthle cell cancer, hypernephroid cancer, infantile embryonal cancer, islet cell cancer, juvenile carcinoma, juvenile nasopharyngeal cancer, Krompecher's cancer, Kupffer cell cancer, laryngeal cancer, lipomatous cancer, lymphoepithelial cancer, lymphoid cancer, mammary cancer, medullary cancer, medulloepithelioma, melanotic cancer, merkel cell cancer, mesonephric cancer, mixed cell cancer, mucinous cancer, myxomatous cancer, nasopharyngeal cancer, oat cell cancer, oncocytic cancer, oncocytoma cancer, osteocarcinoma, ovarian cancer, Paget's cancer, papillary cancer, papillary cystic cancer, paraganglio cancer, parathyroid cancer, pheochromocytoma cancer, pinealoma cancer, pituicytic cancer, pleuropulmonary cancer, polycythemia vera cancer, prickle cell cancer, primary brain cancer, primary carcinoma, primary liver cancer, primary skin cancer, prostatic cancer, rectal cancer, renal cancer, retinoblastoma cancer, Rous sarcoma cancer, sarcoma cancer, scirrhous cancer, sebaceous cancer, small cell cancer, small cell lung cancer, small cell oat cell cancer, small cell with features intermediate between lymphoma and carcinoma cancer, small cell with features intermediate between lymphoma andcarcinoma), breast cancer, inflammatory cancer of the breast, cancer in situ, intraductal cancer of the breast, intraepidermal cancer, intraepithelial cancer, juvenile embryonal cancer, Kulchitsky-cell cancer, large cell cancer, leptomeningeal cancer, lobular cancer, infiltrating lobular cancer, invasive lobular cancer, lobular cancer in situ (LCIS), lymphoepithelial cancer, cancer medullare, medullary cancer, medullary cancer of thyroid gland, medullary thyroid cancer, melanotic cancer, meningeal cancer, Merkel cell cancer, metatypical cell cancer, micropapillary cancer, mucinous cancer, cancer muciparum, cancer mucocellulare, mucoepidermoid cancer, cancer mucosum, mucous cancer, nasopharyngeal cancer, neuroendocrine cancer of the skin, noninfiltrating cancer, non-small cell cancer, non-small cell lung cancer (NSCLC), oat cell cancer, cancer ossificans, osteoid cancer, Paget’s cancer, papillary cancer, papillary cancer of thyroid gland, periampullary cancer, preinvasive cancer, prickle cell, primary intrabony cancer, primary mucosal cancer, prostatic cancer, rectal cancer, renal cancer, reticuloendothelial cancer, retinoblastoma, rhabdomyosarcoma, sarcoma, scirrhous cancer, small cell cancer, small cell lung cancer (SCLC), small cell neuroendocrine cancer, squamous cell cancer, squamous cell carcinoma of the lung, squamous cancer, squamous epithelial cancer, stomach cancer, supratentorial primitive neuroectodermal tumor (sPNET), superficial spreading cancer, synovial sarcoma, T-cell cancer, testicular cancer, thyroid cancer, transitional cell cancer, trophoblastic cancer, tumor, tumor cancer, ulcerating cancer, undifferentiated cancer, undifferentiated carcinoma, undifferentiated sarcoma, urethral cancer, urethral cancer, uterine cancer, vaginal cancer, and vulvar cancer.Intraosseous carcinoma, renal cell carcinoma, scar carcinoma, schistosomal bladder carcinoma, Schneiderian carcinoma, scirrhous carcinoma, sebaceous carcinoma, signet-ring cell carcinoma, simplex carcinoma, small cell carcinoma, small cell lung cancer (SCLC), spindle cell carcinoma, spongiosum carcinoma, squamous carcinoma, squamous cell carcinoma, terminal duct carcinoma, anaplastic thyroid carcinoma, follicular thyroid carcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, trabecular cancer of the skin. Skin, transitional cell carcinoma, tubular carcinoma, undifferentiated thyroid carcinoma, uterine corpus carcinoma, verrucous carcinoma, villous carcinoma, choriocarcinoma, yolk sac carcinoma, squamous cell carcinoma, especially head and neck squamous cell carcinoma, esophageal squamous cell carcinoma, and / or oral cancer or oral cavity cancer.

[0339] Gastrointestinal cancers that can be treated include extrahepatic bile duct cancer, bowel cancer, colon cancer, colon and rectum cancer, colorectal cancer, gallbladder cancer, gastric cancer, gastrointestinal carcinoid tumour, gastrointestinal carcinoid tumour, gastrointestinal stromal tumour, bladder cancer, islet cell cancer (endocrine pancreas), pancreatic cancer, islet cell pancreatic cancer, prostate cancer, rectal cancer, salivary gland cancer, small bowel cancer, colon cancer, and polyps associated with colorectal neoplasia. Lung and respiratory cancers that can be treated include bronchial adenoma / carcinoid, esophagus cancer, esophageal cancer, esophageal cancer, hypopharyngeal cancer, laryngeal cancer, hypopharyngeal cancer, lung carcinoid tumour, non-small cell lung cancer, small cell lung cancer, small cell carcinoma of the lungs, mesothelioma, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, nasopharyngeal cancer, oral cavity cancer, oral cavity and lip cancer, oropharyngeal cancer, paranasal sinus and nasal cavity cancer, and / or pleuropulmonary blastoma.

[0340] Hormonal cancers that can be treated include parathyroid cancer, pineal and supratentorial primitive neuroectodermal tumours, pituitary tumour, thymoma and thymic carcinoma, thymoma, thymus cancer, thyroid cancer, adrenocortical cancer, and / or ACTH-producing tumours.

[0341] Leukemias that can be targeted (which can also be considered to include other blood cell malignancies) include acute lymphoblastic, acute myeloid, acute lymphocytic, acute myelogenous leukemia, chronic myelogenous leukemia, hairy cell leukemia, erythroleukemia, lymphoblastic leukemia, myeloid leukemia, lymphocytic leukemia, myelogenous leukemia, leukemia, hairy cell leukemia, T-cell leukemia, monocytic leukemia, myeloblastic leukemia, granulocytic leukemia, gross leukemia, hand mirror-cell leukemia, basophilic leukemia, hemoblastic leukemia, histiocytic leukemia, leukopenic leukemia, lymphatic leukemia, Schilling's leukemia, stem cell leukemia, myelomonocytic leukemia, monocytic leukemia, prolymphocytic leukemia, promyelocytic leukemia, micromyeloblastic leukemia, megakaryoblastic leukemia, megakaryocytic leukemia, rieder cell leukemia, bovine leukemia, aleukemic leukemia, mast cell leukemia, myelocytic leukemia, plasma cell leukemia, subleukemic leukemia, multiple myeloma, nonlymphocytic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma (indolent and high-grade forms), multiple myeloma, Waldenstrom's macroglobulinemia, myelodysplastic syndrome, and myelofibrosis.Macroglobulinemia), heavy chain disease, myelodysplastic syndrome, and myelodysplasia and / or chronic myelocytic leukemia.

[0342] Hepatic cancers that can be targeted include extrahepatic bile duct cancer and / or hepatocellular carcinoma.

[0343] AIDS-related lymphoma, non-Hodgkin's lymphoma, Hodgkin's lymphoma, T-cell lymphoma, T-cell leukemia / lymphoma, African lymphoma, B-cell lymphoma, B-cell monocytoid lymphoma, bovine malignant lymphoma, Burkitt's lymphoma, centrocytic lymphoma, lymphoma cutis, diffuse lymphoma, diffuse large cell lymphoma, diffuse mixed small and large cell lymphoma, diffuse, small cleaved cell lymphoma, follicular lymphoma, follicular center cell lymphoma, follicular mixed small cleaved and large cell lymphoma, follicular, predominantly large cell lymphoma, follicular, predominantly small cleaved cell lymphoma, giant follicle lymphoma, giant follicular lymphoma, granulomatous lymphoma, histiocytic lymphoma, large cell lymphoma, immunoblastic lymphoma, large cleaved cell lymphoma, large non-cleaved cell lymphoma, Lennert's lymphoma, lymphoblastic lymphoma, lymphocytic, intermediate lymphoma, intermediately differentiated, lymphocytic, small lymphocytic lymphoma, lymphoma, lymphomatoid granulomatosis, mediastinal (thymic) large cell (ML) lymphoma, mixed cell (small and large cell) lymphoma, nodular lymphoma, nodular mixed small and large cell lymphoma, small cell (center cell) lymphoma, small lymphocytic lymphoma, T-cell leukemia / lymphoma, T-cell rich (B-cell) lymphoma, and Waldenstrom's macroglobulinemia.lymphoma, poorly differentiated lymphocytic lymphoma, small lymphocytic lymphoma, well differentiated lymphocytic lymphoma, bovine lymphoma, MALT, mantle cell lymphoma, mantle zone lymphoma, marginal zone lymphoma, Mediterranean lymphoma, lymphoma mixed lymphocytic-histiocytic, nodular lymphoma, plasmacytoid lymphoma, pleomorphic lymphoma, primary central nervous system lymphoma, primary effusion lymphoma, small B-cell lymphoma, small cleaved cell lymphoma, small noncleaved cell lymphoma, T-cell lymphomas, convoluted T-cell lymphoma, cutaneous T-cell lymphoma, small lymphocytic T-cell lymphoma, undefined lymphoma, u-cell lymphoma, undifferentiated, AIDS-related central nervous system lymphoma, cutaneous T-cell lymphoma, effusion (body cavity based) lymphoma, thymic lymphoma lymphoma, and / or cutaneous T-cell lymphoma.

[0344] Suitable proliferative and / or malignant diseases include sarcomas and fibrosarcomas, which are tumors that embed their cells in a fibrous or homogeneous material, such as embryonic connective tissue. Sarcomas that can be targeted include adipose sarcoma, alveolar soft part sarcoma, ameloblastic sarcoma, avian sarcoma, botryoid sarcoma, botryoides sarcoma, chicken sarcoma, chloromatous sarcoma, chondroblastic sarcoma, clear cell sarcoma of tendon sheaths, clear cell sarcoma of kidney, embryonal sarcoma, endometrial stromal sarcoma, epithelioid sarcoma, Ewing’s sarcoma, fascial sarcoma, fibroblastic sarcoma, fowl sarcoma, giant cell sarcoma, granulocytic sarcoma, hemangioendothelial sarcoma, Hodgkin’s sarcoma, idiopathic multiple pigmented hemorrhagic sarcoma, immunoblastic sarcoma of B cells, immunoblastic sarcoma of T cells, Jensen’s sarcoma, Kaposi’s sarcoma, kupffer cell sarcoma, leukocytic sarcoma, lymphatic sarcoma, melanotic sarcoma, mixed cell sarcoma, multiple sarcoma, lymphangio sarcoma, idiopathic hemorrhagic sarcoma, multipotential primary sarcoma of bone, myogenic sarcoma, myxosarcoma, myxomatous sarcoma, myxosarcoma, neurogenic sarcoma, osteogenic sarcoma, palmar fibromatosis, paraganglioma, parosteal sarcoma, plexosarcoma, pleuropulmonary sarcoma, rhabdomyosarcoma, reticulocytic sarcoma, reticulosis, reticulosis sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma, reticulohistiocytic sarcoma,sarcoma (osteosarcoma) / malignant fibrous histiocytoma of bone, and / or soft tissue sarcomas.

[0345] Skin cancers (including non-melanoma) and melanomas that can be treated include cutaneous T-cell lymphoma, intraocular melanoma, metastatic melanoma, tumour progression of human skin keratinocytes, basal cell carcinoma, and squamous cell cancer. Ocular cancers that can be targeted include intraocular melanoma, retinoblastoma, and / or intraocular melanoma.

[0346] Urinary and reproductive system cancers that can be treated include cervical cancer, endometrial cancer, ovarian epithelial cancer, extragonadal germ cell tumour, extracranial germ cell tumour, extragonadal germ cell tumour, ovarian germ cell tumour, gestational trophoblastic tumour, spleen cancer, kidney cancer, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumour, ovarian low malignant potential tumour, penile cancer, renal cancer, retin renal cell cancer (including carcinomas), renal cell cancer, renal pelvis and ureter (transitional cell cancer), transitional cell cancer of the renal pelvis and ureter, gestational trophoblastic tumour, testicular cancer, ureter and renal pelvis, transitional cell cancer of the ureter and renal pelvis, urethral cancer, uterine cancer, and vaginal cancer.transitional cell cancer), urethral cancer, endometrial uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, ovarian carcinoma, primary peritoneal epithelial neoplasms, cervical carcinoma, uterine cancer, solid tumours in the ovarian follicle, superficial bladder tumours, invasive transitional cell carcinoma of the bladder and / or muscle-invasive bladder cancer.

[0347] Various other cancers that can be targeted include advanced cancers, AIDS-related cancers, anal cancer, adrenal cortical cancer, aplastic anemia, aniline-induced cancer, betel nut-induced cancer, bone cancer, buccal mucosa cancer, carcinoids (including gastrointestinal and bronchial carcinoids), Castleman's disease, chronic myeloproliferative disorders, cerebriform carcinoma, chimney-sweeps carcinoma, clay pipe carcinoma, colloid carcinoma, contact carcinoma, cystic carcinoma, dendritic carcinoma, cancer deux, duct carcinoma, and dye worker carcinoma. Workers' cancer, encephaloid cancer, carcinoma en cuirasse, endometrial cancer, endothelial cancer, epithelial cancer, Ewing's family of tumors, glandular cancer, head and neck cancer, hemangiopericytoma, carcinoma in situ, kang cancer, kangri cancer, latent cancer, lip and oral cavity cancer, medullary cancer, melanotic tumor, metastatic squamous neck cancer with occult primary, multiple endocrine neoplasia syndrome, multiple myeloma / plasma cell tumor. neoplasm, mule-spinner's cancer, mycosisSuitably, the proliferative and / or malignant disease is bladder cancer, brain cancer, breast cancer, hepatocellular carcinoma, acute myeloid leukaemia, lung cancer, non-small cell lung cancer, non-Hodgkin's lymphoma, glioma, glioblastoma, melanoma, metastatic melanoma, ovarian cancer or prostate cancer.

[0348] Suitably, the proliferative and / or malignant disease is bladder cancer, brain cancer, breast cancer, hepatocellular carcinoma, acute myeloid leukaemia, lung cancer, non-small cell lung cancer, non-Hodgkin's lymphoma, glioma, glioblastoma, melanoma, metastatic melanoma, ovarian cancer or prostate cancer.

[0349] Suitably, the compound of formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof, is administered alone or with one or more additional therapeutic agents either simultaneously, sequentially or separately.

[0350] Suitably, the one or more additional therapeutic agents are selected from one or more immunotherapeutic agents or other anti-cancer agents.

[0351] Suitably, the immunotherapeutic agent can include one or more cancer vaccines and / or therapeutic antibodies and / or small molecules, and / or RNA / DNA-based agents, and / or viral agents (e.g., oncolytic viruses or oncolytic agents) and / or cell therapies or genetically engineered cell therapies. Cancer vaccines are agents, cell-based agents, molecules, or immunogens in which an endogenous immune response against one or more tumor antigens is stimulated, harnessed, or elicited in an individual or subject.

[0352] Cell therapies or genetically engineered cell therapies include adoptive cell therapies, such as TIL therapies, CAR-T therapies, and other immune effector cells. In some embodiments, the immune effector cells comprise 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.

[0353] As used herein, tumor antigens are broadly defined as antigens that are expressed within or specifically by tumor or cancer cells. Antigens can be expressed on the cell surface, where they are recognized by components of the humoral immune system, such as B lymphocytes (B cells). Intracellular tumor antigens, including those associated with secreted proteins, cytoplasmic proteins, and nuclear proteins, are processed into shorter peptide fragments that form complexes with major histocompatibility complex (MHC) molecules I and II (also known as human leukocyte antigens (HLAs)) and are presented on the cell surface of cancer cells, where they are recognized by T cell receptors (TCRs) of T lymphocytes (T cells). Antigens can also include foreign and self lipids (such as those presented on CD1 molecules) or aberrant glycosylation or post-translational modifications. Preferably, tumor antigens are antigens that are not expressed by normal cells, or at least not to the same level as in tumor cells. Cancer vaccines are currently being developed to activate the humoral immune system (i.e., antibody-dependent immune responses) or the cell-mediated immune system in humans, which includes T lymphocytes (T cells) capable of specifically recognizing and killing tumor cells.

[0354] Cancer vaccines can enhance the presentation of one or more tumor antigens to antigen-presenting cells (e.g., macrophages and dendritic cells) and / or to other immune cells (e.g., T cells, B cells, and NK cells). In some embodiments, the formulation and / or formulation of the cancer vaccine can be used with one or more adjuvants well known in the art to induce or increase an immune response.

[0355] An adjuvant is a substance that is incorporated into or administered with an antigen to enhance the immune response. Adjuvants can enhance the immune response by providing an antigen depot (extracellular or intramacrophage / DC), or by activating antigen presenting cells to stimulate specific lymphocyte collections. Many types of adjuvants are known in the art. Specific examples of adjuvants include monophosphoryl lipid A (MPL, SmithKline Beecham), a homolog obtained after purification and acid hydrolysis of Salmonella Minnesota Re595 lipopolysaccharide; saponins, which include QS21 (SmithKline Beecham), a purified QA-21 saponin from Quillaja Saponaria extract; DQS21, described in PCT application WO 96 / 33739 (SmithKline Beecham); QS-7, QS-17, QS-18 and QS-L1 (So et al., MoI Cells (1997) 7: 178-186); ISCOMATRIX adjuvant, a cage-like structure composed of saponin, phospholipid and cholesterol (see, e.g., Maraskovsky et al., Clin. Cancer Res. (2004) 10: 2879-2890); incomplete Freund's adjuvant; complete Freund's adjuvant; montanide; alum; CpG oligonucleotides (see, e.g., Kreig et al., Nature 374:546-9, 1995) and other immunostimulatory oligonucleotides, including poly-IC and poly-ICLC and various water-in-oil emulsions prepared from biodegradable oils, such as squalene and / or tocopherol.

[0356] Suitably, the cancer cells in the individual express a tumor antigen that is cross- reactive with the cancer vaccine immunization. Suitably, the tumor antigen is expressed in the cancer cells but not in normal somatic cells of the individual. Suitable tumor antigens for use in immunotherapeutic agents (e.g., 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, LB33 / MUM-1, FRAME, NAG, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), tyrosinase, brain type 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, and immunogenic fragments thereof.Other types of tumour antigens include overexpressed or mutated proteins as well as differentiation antigens, especially melanocyte differentiation antigens, such as p53, ras, CEA, MUC1, PMSA, PSA, tyrosinase, Melan-A, MART-1, gp100, gp75, alpha-actinin-4, Bcr-Abl fusion protein, Casp-8, beta-catenin, cdc27, cdk4, cdkn2a, coa-1, dek-can fusion protein, EF2, ETV6-AML1 fusion protein, LDLR-fucosyltransferase AS fusion protein, HLA-A2, HLA-A1 1, hsp70-2, KIAA0205, Mart2, Mum-2 and Mum-3, neo-PAP, class I myosins, OS-9, pml-RAR.alpha fusion protein, PTPRK, K-ras, N-ras, Triosephosphate isomerase, GnTV, Herv-K-mel, NA-88, SP17, TRP2-Int2, (MART-I), E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein-Barr virus antigens, EBNA, human papillomavirus (HPV) antigens E6 and E7, TSP-180, MAGE-4, MAGE-5, MAGE-6, p185 erbB2, p180 erbB-3, c-met, nm-23 H1, PSA, TAG-72-4, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, alpha-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 / cyclophilin C-associated protein), TAAL6, TAG72, TLP, and TPS as well as tyrosinase-related proteins such as TRP-1 and TRP-2.Other suitable antigens include cancer antigens of the following classes: cancer-testis antigens (e.g., HOM-MEL-40), differentiation antigens (e.g., HOM-MEL-55), overexpressed gene products (HOM-MD-21), mutated gene products (NY-COL-2), splice variants (HOM-MD-397), splice peptides (13, 14, 15), gene amplification products (HOM-NSCLC-11), and self-antigens associated with cancer (HOM-MEL-2.4), as described in Cancer Vaccines and Immunotherapy (2000) Eds Stern, Beverley and Carroll, Cambridge University Press, Cambridge. 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, melan-A, and mixtures thereof. In some variations, the cancer antigen is a mammalian protein. In some variations, the cancer antigen is a human protein. In some variations, a full-length protein can be used as an antigen. In some variations, a peptide comprising an antigenic fragment of these proteins can be used as a tumor antigen.

[0357] Other suitable tumor antigens are well known in the art (see, e.g., WO 00 / 20581). The sequences of these tumor antigens are readily available from public databases, but are also found in WO 1992 / 020356, WO 1994 / 005304, WO 1994 / 023031, WO 1995 / 020974, WO 1995 / 023874, and WO 1996 / 026214.

[0358] In some embodiments, the methods of treating a subject with a compound of Formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof, and an immunotherapeutic agent as disclosed herein can further comprise administering one or more tumor therapies to treat the tumor. Such therapies include, for example, tumor medicaments, radiation, and surgery.

[0359] As used herein, “tumor medicament” refers to an agent that is administered to a subject for the purpose of treating cancer. Various types of medicaments for treating tumors are described herein.

[0360] Tumor drugs exert their effects in a variety of ways. Some cancer drugs exert their effects by targeting physiological mechanisms specific to tumor cells. Examples include targeting specific genes and their gene products (i.e., primarily 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). Tumor drugs can alternatively target signal transduction pathways and other molecular mechanisms that are altered in tumor cells.

[0361] Immunotherapeutic agents used in combination with a compound as described herein can include biological response modifiers. Biological response modifiers for use in the combination therapies disclosed herein can include: interferon-alpha, IL-2, G-CSF, and GM-CSF; a variety of agents including platinum coordination complexes such as cisplatin and carboplatin, anthracenediones such as mitoxantrone, substituted ureas such as hydroxyurea, methylhydrazine derivatives including N-methylhydrazine (MIH) and procarbazine, adrenocortical suppressants such as mitotane (o,p'-DDD) and aminoglutethimide; hormones and antagonists including adrenocorticosteroid antagonists such as prednisone and its equivalents, dexamethasone, and aminoglutethimide; progestins such as hydroxyprogesterone caproate, medroxyprogesterone acetate, and megestrol acetate; estrogens such as diethyl stilbestrol and ethinyl estradiol equivalents; antiestrogens such as tamoxifen; androgens including testosterone propionate and fluoxymesterone and equivalents; antiandrogens such as flutamide, gonadotropin-releasing hormone analogs, and leuprolide; non-steroidal antiandrogens such as flutamide;Kinase inhibitors, histone deacetylase inhibitors, methylation inhibitors, proteasome inhibitors, oxidants, antioxidants, telomerase inhibitors, BH3 mimetics, ubiquitin ligase inhibitors, stat inhibitors; receptor tyrosine kinase inhibitors, such as Imatinib (Gleevec or Glivec) and Erlotinib (an EGF receptor inhibitor, marketed as Tarceva).

[0362] Additional biological response modifiers can include inhibitors of the immune modulating enzyme indoleamine 2,3-dioxygenase (IDO or IDO or INDO EC 1.13.11.52), tryptophan 2,3-dioxygenase (TDO, EC 1.13.11.11).

[0363] Tryptophan 2,3-dioxygenase (TDO) is a homotetrameric heme-containing cytosolic enzyme encoded by the gene TD02 and expressed at high levels in the liver. It catalyzes the first and rate-limiting step of tryptophan degradation along the kynurenine pathway, thereby regulating systemic tryptophan levels. The work of Pilotte et al. (Proc Natl Acad Sci U S A. 2012 Feb 14; 109(7): 2497-502. Epub 2012 Jan 30) demonstrates the role of TDO in cancer. These studies detected TDO expression in a significant proportion of human tumors. In preclinical models, TDO expression by tumors prevented immune mice from rejecting the tumors. These studies used a TDO inhibitor, which, after systemic treatment, restored the ability of mice to reject tumors expressing TDO, providing evidence that TDO inhibitors are effective in cancer treatment. TDO inhibitors include LM10.

[0364] Indoleamine 2,3 dioxygenase (IDO) is an enzyme encoded in humans by the IDOl gene. This enzyme also catalyzes the first and rate-limiting step in the degradation of the essential amino acid L-tryptophan to N-formylkynurenine, and is normally expressed in tumor cells and activated immune cells. IDO suppresses the immune response by degrading the indole moiety of tryptophan, locally depleting tryptophan levels and increasing proapoptotic kynurenine. Thus, IDO blocks the proliferation and activation of T cells, which are extremely sensitive to Trp deficiency. This creates an environment in which tumor-specific cytotoxic T lymphocytes are functionally inactivated or no longer able to attack the patient's cancer cells. The observation that many human tumors constitutively express IDO introduced the hypothesis that inhibition of IDO can enhance the effectiveness of cancer immunotherapy. Results from in vitro and in vivo studies suggest that the efficacy of therapeutic vaccination of cancer patients can indeed be improved by concomitant administration of IDO inhibitors.

[0365] Small molecule inhibitors of IDO are available in the art for the treatment of IDO related diseases, such as cancer. For example, W099 / 29310 reports methods for altering T cell mediated immunity comprising altering local extracellular concentrations of tryptophan and tryptophan metabolites, etc. Other compounds having IDO inhibitory activity are reported in WO2004 / 094409, US8088803 (which reports the INCB024360 compound), US20110165188 and US20110159017. Immunotherapeutic agents for use in combination with the compounds as described herein can include cytokines.

[0366] 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), TNF alpha, TNF1, TNF2, G-CSF, Meg-CSF, GM-CSF, thrombopoietin, stem cell factor, and erythropoietin.

[0367] As used herein, the term "immunotherapy," "immunotherapeutic agent," or "immunotherapeutic" generally refers to any therapeutic approach that aims to mobilize or manipulate the immune system of a patient to treat or cure a disease, in particular a disease such as cancer. The term "immunotherapy," "immunotherapeutic agent," or "immunotherapeutic" includes targeting tumor cells by recognizing immunogenic proteins or antigens expressed by said tumor cells, either by exploiting passively transferred immune molecules (e.g., antibodies) or by exploiting cancer vaccine preparations designed to induce antibodies or T lymphocytes (T cells) that recognize specific regions or epitopes of tumor cell-specific antigens.

[0368] The immunotherapeutic agent used in combination with the compound as described herein can be an antibody.

[0369] Suitably, the therapeutic antibody comprises one or more of an anti-Her2 / neu receptor antibody, such as trastuzumab (trade name Herceptin); Alemtuzumab (a CD52 antibody, trade name Campath, MabCampath or Campath-1H, currently being further developed under the name Lemtrada); Gemtuzumab (an anti-CD33 monoclonal antibody linked to calicheamicin, marketed by Wyeth under the trade name Mylotarg); an anti-CD20 antibody, such as Rituximab (trade name Rituxan and MabThera) or Ibritumomab tiuxetan (sold under the trade name Zevalin); an anti-TNF-alpha antibody, such as Infliximab (trade name Remicade) or Adalimumab (trade name Humira), or a soluble TNFR2 molecule, such as etanercept (also known as Enbrel); an antibody against the CD25 chain of the IL-2 receptor, such as basiliximab (trade name Simulect); an anti-CD40 / CD40L antibody, such as humanized IgGl anti-human CD40 antibody (SGN-40); an anti-CTLA-4 blocking antibody, such as Ipilimumab (also known as MDX-101 or MDX-010, trade name Yervoy) or tremelimumab; an anti-PD-1 antibody (programmed cell death protein 1, also known as CD279); an anti-PD-L1 antibody (programmed cell death ligand); an antibody against a glucocorticoid-induced TNFR family-related gene, or an anti-GITR antibody; or an anti-OX-40 (CD134) antibody.

[0370] Other suitable immunotherapeutic agents include immunomodulatory agents based on soluble lymphocyte activation gene 3 (also known as LAG3 or CD223), e.g., LAG3-Ig (IMP321); Toll-like receptor agonists, e.g., MPL, CpG, single-stranded RNA, nucleotides, nucleotide analogs, CL087 (a TLR7 specific ligand), loxoribine, polyinosine-polycytidylic acid, flagellin, resiquimod, immiquimod, gardiquimod, and NOD ligands, e.g., muramyl dipeptide, murabutide, peptidoglycan, muramyl dipeptide, and antiviral agents, e.g., oseltamivir phosphate, Amphotericin B, and palivizumab.

[0371] In some embodiments, a therapy as disclosed herein can comprise administration of a compound as described herein (e.g., a compound of Formula (II), or (III), or (IV), or (V), (VI), or a salt and solvate thereof) and one or more antibodies selected from the group consisting of an anti-PD1 antibody, an anti-PDL-1 antibody, an anti-CTLA-4 antibody, an anti-GITR antibody, and an anti-OX40 antibody.

[0372] An anti-PD1 antibody can be a monoclonal antibody directed against the negative immune regulatory human cell surface receptor PD-1 with immune-enhancing activity. An exemplary anti-PD1 antibody is the human monoclonal antibody MDX-1106, which binds to PD-1 and blocks the activation of PD-1 by its ligands PD-L1 and PD-L2, thereby activating T cells and cell-mediated immune responses against tumor cells. In some embodiments, an anti-PD-1 antibody is a monoclonal antibody directed against the protein ligand PD-L1 with immune regulatory and anti-tumor activity. An exemplary anti-PD-L1 antibody is the human monoclonal antibody MDX-1105, which binds to PD-L1 and blocks the binding and activation of PD-1 to its receptor PD-1, thereby enhancing T cell-mediated immune responses against tumors and reversing T cell inactivation in chronic infectious diseases. PD-L1 is widely expressed on hematopoietic and parenchymal tissues.

[0373] The anti-CTLA-4 antibody can be a monoclonal antibody directed 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 the antigen presenting cell ligands B7-1 and B7-2 to CTLA-4, resulting in the inhibition of downregulation of B7-CTLA4-mediated T-cell activation. Another exemplary anti-CTLA-4 antibody is the human IgGl monoclonal antibody ipilimumab, which binds to CTLA4 and blocks the binding of the antigen presenting cell ligands B7-1 and B7-2 to CTLA-4, resulting in the inhibition of downregulation of B7-CTLA4-mediated T-cell activation. Ipilimumab is undergoing clinical trials for the treatment of non-small cell lung cancer, small cell lung cancer, and metastatic hormone-refractory prostate cancer.

[0374] The anti-GITR antibody can be a monoclonal antibody directed against the glucocorticoid-induced tumor necrosis factor receptor (GITR) that blocks the interaction of GITR with its ligand, enhances the cytotoxicity of human natural killer cells, and / or downregulates GITR expression on peripheral blood lymphocytes.

[0375] The anti-OX40 antibody can be an agonistic monoclonal antibody that mimics the natural OX40 ligand and selectively binds to and activates the OX40 receptor. Receptor activation induces proliferation of memory and effector T cells.

[0376] Suitably, the anti-cancer agent can include any known agent having the desired anti-cancer properties. Suitably, the anti-cancer agent includes one or more of the following agents: taxoids, such as paclitaxel (Taxol®) and docetaxel (Taxotere®); platinum coordination complexes, such as cisplatin, carboplatin, and oxaliplatin; alkylating agents, such as mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, and nitrogen mustard; antimetabolites, such as methotrexate, 5-fluorouracil, and cytosine arabinoside; antibiotics, such as dactinomycin (actinomycin D), daunorubicin, doxorubicin, idarubicin, bleomycin, plicamycin (mithramycin), and mitomycin; enzymes, such as L-asparaginase; topoisomerase inhibitors, such as aclarubicin, amsacrine, etoposide, etoposide phosphate, teniposide, and topotecan; DNA synthesis inhibitors, such as fluorouracil and tegafur; DNA cross-linking agents, such as busulfan, carmusta, die, lomustine, and streptozocin; DNA synthesis and / or repair inhibitors, such as hydroxyurea; and biological response modifiers, such as interferon-alpha. albumin paclitaxel (Abraxane) or other chemotherapeutic agents such as cis-platin and other platin intercalating compounds, etoposide and etoposide phosphate, bleomycin, mitomycin C, CCNU, doxorubicin, daunorubicin, idarubicin, ifosfamide, and the like. Other anticancer agents that can be contemplated for use in the combination therapies disclosed herein include aspirin, sulindac, curcumin, alkylating agents, including: nitrogen mustards, such as mechlorethamine, cyclophosphamide, ifosfamide, melphalan, and chlorambucil; nitrosoureas, such as carmustine (BCNU), lomustine (CCNU), and semustine (methyl-CCNU); ethylenimines / methylmelamines, such as thriethylenemelamine (TEM), triethylene thiophosphoramide (thiotepa), hexamethylmelamine (HMM, altretamine); alkyl sulfonates, such as busulfan; triazines, such as dacarbazine (DTIC); antimetabolites, including: folic acid analogs, such as methotrexate and trimetrexate;pyrimidine analogs such as 5-fluorouracil, fluorodeoxyuridine, gemcitabine, cytosine arabinoside (Ara C, cytarabine), 5-azacytidine, 2,2'-difluorodeoxycytidine; purine analogs such as 6-mercaptopurine, 6-thioguanine, azathioprine, 2'-deoxycoformycin (pentostatin), erythrohydroxynonyladenine (EHNA), fludarabine phosphate, 2-chlorodeoxyadenosine (cladribine, 2-CdA); natural products including: antimitotic drugs such as paclitaxel, vinca alkaloids including vinblastine (VLB), vincristine, vinorelbine, taxotere, estramustine, estramustine phosphate; epipodophylotoxins such as etoposide and tenyposide; antibiotics such as actimomycin D, daunomycin (rubidomycin), doxorubicin, mitoxantrone, idarubicin, bleomycins, plicamycin (mithramycin), mitomycin C, actinomycin;enzymes, such as L-asparaginase. Anti-cancer agents can also be biological agents, such as proteins that inhibit tumor growth, such as interferons (IFN-γ), tumor necrosis factor (TNF-α), TNF-β, GM-CSF, and similar cytokines; or anti-angiogenic factors, such as angiostatin and endostatin; or inhibitors of FGF or VEGF, such as soluble forms of angiogenic factor receptors, including soluble VGF / VEGF receptors. Other anti-cancer agents can also be platinum coordination complexes, such as cisplatin and carboplatin, anthracenediones, such as mitoxantrone, substituted ureas, such as hydroxyurea, methylhydrazine derivatives, which include N-methylhydrazine (MIH) and procarbazine, adrenocortical suppressants, such as mitotane (o,p'-DDD) and aminoglutethimide;Hormones and antagonists, including adrenocorticosteroid antagonists such as prednisone and its equivalents, dexamethasone and aminoglutethimide, progestins such as hydroxyprogesterone caproate, medroxyprogesterone acetate and megestrol acetate, estrogens such as diethyl stilbestrol and its ethinyl estradiol equivalents, antiestrogens such as tamoxifen, androgens including testosterone propionate and fluoxymesterone and their equivalents, antiandrogens such as flutamide, gonadotropin-releasing hormone analogs and leuprolide, non-steroidal antiandrogens such as flutamide;Kinase inhibitors, histone deacetylase inhibitors, methylation inhibitors, proteasome inhibitors, monoclonal antibodies, oxidants, antioxidants, telomerase inhibitors, BH3 mimetics, ubiquitin ligase inhibitors, STAT inhibitors, receptor tyrosin kinase inhibitors, such as imatinib mesylate (marketed under the trade names Gleevac or Glivac) and erlotinib (an EGF receptor inhibitor, now marketed under the trade name Tarveca); and anti-virals, such as oseltamivir phosphate, Amphotericin B and palivizumab. Also included are STING agonists, such as DMXAA.

[0377] Suitably, the anti-cancer agent comprises one or more pyrimidine analogues and an anti-mitotic drug. More suitably, the anti-cancer agent comprises one or more of: 5-fluorouracil, fluorodeoxyuridine, gemcitabine, AraC, cytarabine, 5-azacytidine, 2,2’-difluorodeoxycytidine, paclitaxel, vinblastine (VLB), vincristine, vinorelbine, taxotere, estramustine and estramustine phosphate.

[0378] Suitably, the anti-cancer agent can also comprise an antibody directly or indirectly linked to one or more agents having a desired anti-cancer property to form an antibody-drug conjugate. Suitable antibodies can be selected that will direct the conjugate to cells of interest, such as tumor cells. Such antibody-drug conjugates are well known in the art (16).

[0379] Any type of cell can be treated, including but not limited to bone cells, eye cells, head and neck cells, lung cells, gastrointestinal cells (including, for example, oral, esophageal, intestinal, colon), breast / mammary cells, cervical cells, ovarian cells, uterine cells, prostate cells, liver / hepatic cells, kidney / renal cells, bladder cells, pancreatic cells, brain cells, and skin cells.

[0380] The skilled person will readily be able to determine whether a candidate compound treats a proliferative and / or malignant condition of any particular cell type.

[0381] Suitable subjects are humans, livestock, and companion animals. More suitably, the subject is a human.

[0382] Anti-metastatic

[0383] It has been found that a compound of formula (I), or a metal derivative, salt and solvate thereof, wherein the compound is a compound of formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof, such as a compound of formula (VII), or (VIII), or (IX), or (X), or (XI), or a salt and solvate thereof, has anti-metastatic activity.

[0384] Accordingly, the use of a compound of formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof, in the manufacture of a medicament for treating or inhibiting metastasis can be found.

[0385] Suitably, the compound of formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof, for use in treating or inhibiting metastasis is a compound of formula (VII), or (VIII), or (IX), or (X), or (XI), or a salt and solvate thereof. More suitably, the compound is a compound of formula (II) or (III), or a salt and solvate thereof. More suitably, the compound is a compound of formula (VII) or (VIII), or a salt and solvate thereof.

[0386] Treating metastasis comprises administering to a subject in need a therapeutically effective amount of a compound as described herein or a pharmaceutical composition as described herein.

[0387] Suitably, the metastasis is derived from a bladder cancer, breast cancer, colorectal cancer, esophageal cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer or stomach cancer or any combination thereof.

[0388] Cancer cell spread from the primary tumor site to distant organs is called metastasis. Metastasis of cancer tumors is a cause of many treatment failures when treating the disease, as the patient succumbs to multiple tumor growths. The degree to which metastasis occurs varies from individual tumor type to individual tumor type. Melanoma, breast cancer, lung cancer and prostate cancer are particularly prone to metastasis.

[0389] When metastasis occurs, secondary tumors can form at multiple sites in the body, one of the more common sites of metastasis being the lung.

[0390] Thus, it is beneficial to inhibit tumor metastasis to any degree, regardless of whether the agent involved has any effect on the primary tumor. However, it is an additional advantage of the agent if it also inhibits the primary tumor.

[0391] The metastasis inhibition disclosed herein can be used alone or in combination as part of a treatment regimen for an animal or human patient having a cancer that is prone to metastasis, particularly melanoma, breast cancer, lung cancer and prostate cancer. It is best to administer the treatment that inhibits metastatic formation as soon as possible after the cancer is detected. By using the treatment regimen in the patient at an early stage, the treating physician maximizes the chance that no significant metastasis has yet occurred. This maximizes the chance of successful treatment. In such a regimen, the anti-metastatic factor or salt thereof can and usually will be administered in combination with another form of therapy that controls the primary tumor itself.

[0392] Suitably, the compound of formula (I) or metal derivative, salt and solvate thereof for use in the treatment or inhibition of metastasis is administered either simultaneously with or sequentially with one or more immunotherapeutic agents or other anti-cancer agents.

[0393] The treatments described herein can also be used in conjunction with (i.e. before or after) surgical procedures to remove primary tumor material from the body. Typically, surgical procedures to remove tumor material from the body are avoided due to concerns that the physical manipulation involved will cause tumor cells to metastasize. However, if an anti-metastatic agent as described herein is administered to the patient prior to the surgical procedure, the risk of metastasis that can be caused by the surgical procedure can be reduced and the surgical procedure will be a more attractive treatment option.

[0394] Within the scope of sound medical judgment, the dosage and method of administration of the anti-metastatic agents as described herein will vary with the severity and nature of the particular condition being treated, the duration of treatment, the auxiliary therapies used, the age and physical condition of the patient, and similar factors within the knowledge and expertise of the attending physician. However, a single dose can generally be in the range of 0.01 to 2000 mg / kg body weight, preferably 1 to 200 mg / kg (as used herein, units specified as "mg / kg" refer to milligrams per kilogram of body weight, unless otherwise specified). Up to four doses per day can be routinely used, but this can vary according to the needs of the patient, consistent with a reasonable benefit / risk ratio. Variations in the patient's response can be expected, but generally higher doses within the ranges shown will be required in the case of oral administration, while lower doses shown will apply in the case of intravenous administration.

[0395] Pharmaceutical composition

[0396] Suitably, the pharmaceutical composition further comprises one or more additional therapeutic agents.

[0397] Suitably, the pharmaceutical composition further comprises one or more additional therapeutic agents selected from one or more immunotherapeutic agents or other anti-cancer agents.

[0398] Suitably, the pharmaceutical composition further comprises one or more other anti-cancer agents.

[0399] Suitably, the pharmaceutical composition further comprises one or more cancer vaccines or therapeutic antibodies. Suitably, the pharmaceutical composition further comprises an adjuvant.

[0400] Suitably, the pharmaceutical composition comprises a compound of formula (II), or (III), or (IV), or (V), or (VI), or a salt and solvate thereof, a cancer vaccine or therapeutic antibody, an adjuvant, and a pharmaceutically acceptable excipient, carrier or diluent.

[0401] Suitably, the pharmaceutical composition is formulated as a nanoparticle carrier vehicle or a viral vector.

[0402] Suitably, the pharmaceutical composition is formulated as a nanoparticle carrier vehicle. Suitably, the nanoparticle carrier vehicle is one of a silica nanoparticle, a liposome, a micelle, a nanogel or a polymeric nanoparticle. Suitably, the nanoparticle carrier vehicle is a liposome.

[0403] Suitably, the pharmaceutical composition is formulated as a viral vector. Suitably, the viral vector is an adenoviral, adeno-associated viral, retroviral, lentiviral or herpes viral vector. The use of viral vectors is a well-known method of delivery (17, 18).

[0404] Administration and dosage

[0405] The compounds of formula (II), or (III), or (IV), or (V), or (VI), or salts and solvates thereof, can be administered alone or in combination with one or another compound of formula (II), or (III), or (IV), or (V), or (VI), or salts and solvates thereof, or with one or more pharmacologically active compounds different from the compounds of formula (II), or (III), or (IV), or (V), or (VI), or salts and solvates thereof.

[0406] The compounds of the application can be suitably combined with various components to prepare compositions of the application. Suitably, the compositions are combined with a pharmaceutically acceptable carrier or diluent to prepare a pharmaceutical composition (which can be for human or animal use). Suitable carriers and diluents include isotonic saline solutions (e.g. phosphate buffered saline), water, ethanol, propylene glycol, glycerol, and combinations thereof. Useful pharmaceutical compositions and methods for their preparation can be found in the standard pharmaceutical literature. See, for example, 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. D. B. Troy) 2006 (Lippincott, Williams and Wilkins, Philadelphia, USA), which are incorporated herein by reference.

[0407] The compounds of the application can be administered by any suitable route. Suitably, the compounds of the application are typically administered orally or by any parenteral route in the form of a pharmaceutical formulation comprising the active ingredient, optionally in the form of a non-toxic organic or inorganic acid or base addition salt, in a pharmaceutically acceptable dosage form. Suitably, the parenteral route is selected from intramuscular, subcutaneous, intravenous and intradermal administration. More suitably, the parenteral route is selected from intramuscular and intravenous administration.

[0408] The compounds of the application, their pharmaceutically acceptable salts and pharmaceutically acceptable solvates of any entity can be administered alone but will generally be administered in admixture with a suitable pharmaceutical excipient, diluent or carrier, selected with regard to the intended route of administration and standard pharmaceutical practice.

[0409] Suitable pharmaceutically acceptable excipients include, but are not limited to, any adjuvant, diluent, excipient, glidant, granulation binder, lubricant, sweetening agent, preservative, dye / colorant, flavor enhancer, surface-active agent, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsor that has been approved by the United States Food and Drug Administration (FDA) for use in humans or domestic animals.

[0410] For example, the compounds of the present application, or salts or solvates thereof, can be administered orally, buccally, or sublingually in the form of tablets, capsules (including soft gel capsules), ovules, elixirs, solutions or suspensions for immediate-, delayed-, modified-, sustained-, controlled- or pulsatile delivery, which can contain flavoring agents or coloring agents, and which can be formulated with binders as in gelatin or sucrose-fatty acid acylate, or with dispersants, as in starch. The compounds of the present application can also be administered rectally in the form of suppositories, pessaries, or ovules for immediate-, delayed-, modified-, sustained-, controlled- or pulsatile delivery.

[0411] Such tablets can contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dicalcium phosphate, and glycine; disintegrants such as starch (preferably corn, potato or tapioca starch), sodium starch glycolate, croscarmellose sodium, and certain complex silicates; and granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin, and acacia. Additionally, lubricating agents such as magnesium stearate, stearic acid, glyceryl behenate, and talc can be included.

[0412] Solid compositions of a similar type can also be employed as fillers in gelatin capsules. Preferred excipients in this regard include lactose, starch, cellulose, milk sugar, or high molecular weight polyethylene glycols. For aqueous suspensions and / or elixirs, the compounds of the present application can be combined with various sweetening or flavoring agents, coloring matter or dyes, emulsifying and / or suspending agents, and diluents such as water, ethanol, propylene glycol, and glycerin, and combinations thereof.

[0413] Modified release and pulsatile release dosage forms can contain excipients such as those detailed for immediate release dosage forms, as well as additional excipients used as release rate modifiers, coated on and / or included in the body of the device. Release rate modifiers include, but are not limited to, hydroxypropyl methylcellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, cellulose acetate, polyvinyl oxides, xanthan gum, carbomer, ammoniomethacrylate copolymer, hydrogenated castor oil, carnauba wax, paraffin wax, cellulose acetate phthalate, hydroxypropylmethyl cellulose phthalate, methacrylic acid copolymer, and mixtures thereof. Modified release and pulsatile release dosage forms can contain one or a combination of release rate modifying excipients. Release rate modifying excipients can be present within the dosage form (i.e., within the matrix) and / or on the dosage form (i.e., on the surface or coating).

[0414] Fast dispersing or dissolving dosage forms (FDDF) can contain the following ingredients: aspartame, acesulfame potassium, citric acid, croscarmellose sodium, crospovidone, diascorbic acid, ethyl acrylate, ethyl cellulose, gelatin, hydroxypropyl methylcellulose, magnesium stearate, mannitol, methyl methacrylate, mint flavouring, polyethylene glycol, fumed silica, silicon dioxide, sodium carboxymethyl starch, sodium stearyl fumarate, sorbitol, xylitol.

[0415] The compounds of the application can also be administered parenterally, e.g., intravenously, intraarterially, or the compounds of the application can be administered by infusion techniques. For such parenteral administration, they are best used in the form of a sterile aqueous solution, which can contain other substances, e.g., enough saline or dextrose to make the solution isotonic with the blood. If desired, the aqueous solution can be suitably buffered (preferably to a pH of 3 to 9). Preparation of a suitable parenteral formulation under sterile conditions is readily accomplished by standard pharmaceutical practice, as is well known to those in the art.

[0416] Suitably, the formulations of the application are optimized for the route of administration (e.g., oral, intravenous), etc.

[0417] Administration can be carried out during the course of treatment, at one dose, continuously or intermittently (e.g. in divided doses at appropriate intervals). Methods for determining the most effective means and dosage are well known to skilled persons and will vary with the formulation used for therapy, the purpose of the therapy, the target cell to be treated and the subject to be treated. Single or multiple administrations can be carried out with the dose level and dose regimen being selected by the treating physician, veterinarian or clinician.

[0418] When more than one agent is to be administered, the agents can be provided by administration simultaneously or sequentially. By "simultaneous" administration is meant that the different agents are administered to the individual at the same point in time. This can be achieved as a single dose by the same route of administration or by different routes of administration occurring at the same time. This can occur, for example, where one agent is administered by infusion or parenteral administration while the other agent is given orally during the infusion or parenteral administration.

[0419] By "sequentially" is meant that the different agents are administered at different points in time, provided that the activity of the first agent administered is present and ongoing in the subject at the time of administration of the second agent. For example, an immunotherapeutic agent can be administered first, such that an immune response is generated against a tumour antigen, and then a compound of formula (II), or (III), or (IV), or (V), or (VI), or a derivative compound thereof, or a salt and solvate thereof, is administered, so as to reduce immune suppression at the tumour site. This sequential administration can occur by the same route of administration or by different routes of administration. Preferably, the sequential dosing occurs such that the second of the two agents is administered within 48 hours, preferably within 24 hours, for example 12, 6, 4, 2 or 1 hour, of the first agent. Preferably, one agent can be administered daily, the second agent can be administered every two days, or every three days, or every four days, or every five days or every six days, or every seven days.

[0420] Multiple doses of the compound of Formula (II), or (III), or (IV), or (V), or (VI), or salt and solvate thereof, can be administered, for example 2, 3, 4, 5, or more than 5 doses, can be administered after administration of one or more immunotherapeutic agents or other anticancer agents. Administration of the compound of Formula (II), or (III), or (IV), or (V), or (VI), or salt and solvate thereof, can continue for a period of time after administration of the immunotherapeutic agent or other anticancer agent. For example, treatment with the compound of Formula (II), or (III), or (IV), or (V), or (VI), or salt and solvate thereof, can continue for at least 1 week, at least 2 weeks, at least 3 weeks, at least 1 month, or at least 2 months. Treatment with the compound of Formula (II), or (III), or (IV), or (V), or (VI), or salt and solvate thereof, can be in cycles of discontinuing and then resuming treatment. Treatment with the compound of Formula (II), or (III), or (IV), or (V), or (VI), or salt and solvate thereof, can continue for as long as necessary to achieve complete tumor rejection.

[0421] Multiple doses of one or more immunotherapeutic agents or other anticancer agents can be administered, for example 2, 3, 4, 5, or more than 5 doses, can be administered after administration of the compound of Formula (II), or (III), or (IV), or (V), or (VI), or salt and solvate thereof. Administration of the immunotherapeutic agent or other anticancer agent can continue for a period of time after administration of the compound of Formula (II), or (III), or (IV), or (V), or (VI), or salt and solvate thereof. For example, treatment with the immunotherapeutic agent or other anticancer agent can continue for at least 1 week, at least 2 weeks, at least 3 weeks, at least 1 month, or at least 2 months. Treatment with the immunotherapeutic agent or other anticancer agent can continue for as long as necessary to achieve complete tumor rejection.

[0422] The composition can be administered in different dosages depending on the condition and patient to be treated and the route of administration. For example, a typical dose for an adult human can be 100 ng to 25 mg (suitably about 1 microgram to about 10 mg) per kg body weight of the subject per day.

[0423] When estimating an initial dose for a human subject, guidance can be obtained from studies in test animals. For example, when determining a particular dose for a mouse, suitably the initial test dose for a human can be about 0.5 to 2 times the mg / Kg value given to the mouse.

[0424] Other forms

[0425] Unless otherwise indicated, the foregoing recitations include the well-known ionic, salt, solvate, tautomeric, and protected forms of these substituents. For example, reference to a carboxylic acid group (-RCOOH) also includes the anionic (carboxylate) form (-RCOO - ), salts or solvates thereof, and the conventional protected forms. Similarly, reference to an amino group includes the protonated form (-RN + HR 1 R 2 ), salts or solvates, e.g., hydrochloride, and the conventional protected forms of the amino group. Similarly, reference to a hydroxyl group also includes the anionic form (-O - ), salts or solvates thereof, and the conventional protected forms.

[0426] Certain compounds can exist in one or more particular geometric, optical, enantiomeric, diastereomeric, epimeric, atropic, mesomeric, stereoisomeric, tautomeric, conformational, or anomeric forms. These forms, while not explicitly listed, are individually included in the present disclosure. Such isomeric forms can be defined, in terms of absolute stereochemistry, as (R)- or (S)-, as (l)- or (d)-, or as (l)- or (l)- for arotic compounds, as (D)- or (L)- for amino acids; as (+) or (-) for a given compound, e.g. (+) or (-) a given chiral compound, e.g. (+) or (-) a given chiral amine, or as (D)- or (L)- for a given compound, e.g. (D)- or (L)- a given chiral compound, e.g. (D)- or (L)- a given chiral amine. Also included are any and all geometric, optical, enantiomeric, diastereomeric, epimeric, atropic, mesomeric, stereoisomeric, tautomeric, conformational, or anomeric forms of the present compounds. For example, if a compound of the present disclosure incorporates a chiral center, it can exist, independently, as either an (R)- or (S)-enantiomer, or mixtures thereof. When a compound of the present disclosure incorporates more than one chiral center (e.g., two chiral centers), then the present disclosure includes all possible enantiomeric and diastereomeric mixtures, as well as the individual (R)- and (S)-enantiomers. Similarly, if a compound of the present disclosure incorporates an aromatic ring, it can exist as either an (E)- or (Z)- geometric isomer, or mixtures thereof. Unless otherwise indicated, the present disclosure includes all such possible isomers, including racemic mixtures, optically pure forms, and other mixtures.

[0427] Note that, except as discussed below with respect to tautomeric forms, specifically excluded from the term "isomers" as used herein are structural (or compositional) isomers (i.e., isomers that differ in their connectivity of atoms, rather than merely their position in space). For example, reference to a methoxy group -OCH3should not be interpreted as reference to its structural isomer hydroxymethyl -CH2OH.

[0428] Reference to a class of structures can well include structural isomers falling within the class (e.g., C 1-7 Alkyl includes methyl, ethyl, propyl, isopropyl; butyl, isobutyl, sec-butyl, t-butyl; methoxyphenyl includes o-methoxyphenyl, m-methoxyphenyl, and p-methoxyphenyl.

[0429] The above exclusions do not apply to tautomeric forms, such as keto, enol and enolate forms, for example as shown in the following tautomeric pairs: keto / enol, imine / enamine, amide / imino alcohol, amidine / amidine, nitroso / oxime, thioketone / enethiol, N-nitroso / hyroxyazo, and nitro / aci-nitro.

[0430] Note that the term“isomer” specifically includes compounds that have one or more isotopic substitutions. For example, H can be any isotopic form, including 1 H, 2 H(D), and 3 H(T); C can be any isotopic form, including 12 C, 13 C, and 14 C; O can be any isotopic form, including 16 O, and 18 O; and the like.

[0431] Unless otherwise stated, a reference to a particular compound includes all such isomeric forms, including (wholly or partially) racemic mixtures, where these are otherwise possible.

[0432] Methods for preparing (e.g., asymmetric synthesis) and isolating (e.g., fractional crystallization and chromatographic means) such isomeric forms are known in the art or are readily obtained by modification of the methods taught herein or known methods, by way of known procedures.

[0433] Unless otherwise stated, a reference to a particular compound also includes solvates, salts, and protected forms thereof, for example as described below.

[0434] In some embodiments, the compounds of the present disclosure, and salts and solvates thereof, comprise pharmaceutically acceptable salts of the compounds of the present disclosure.

[0435] The compounds of the present disclosure, including the compounds specifically named above, can form pharmaceutically acceptable complexes, salts, solvates, and hydrates. These salts include nontoxic acid addition salts (including diacids) and base salts.

[0436] If the compound is cationic or has a functional group which can be cationic (e.g., -NH2 can be -NH3+), salts of the compound can be prepared from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfinic, phosphoric, nitric, and the like; or from organic acids such as aliphatic-mono- and -dicarboxylic acids and +When the compound of the application is basic, salts can be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include, by way of example only, acetic, 2- acetyloxybenzoic, ascorbic, aspartic, benzoic, camphorsulfonic, cinnamic, citric, edetic, ethanedisulfonic, ethanesulfonic, fumaric, glucheptonic, gluconic, glutamic, glycolic, hydroxymaleic, hydroxynaphthalenecarboxylic, isethionic, lactic, lactobionic, lauric, maleic, malic, methanesulfonic, mucic, oleic, oxalic, palmitic, pamoic, pantothenic, phenylacetic, phenylsulfonic, propionic, pyruvic, salicylic, stearic, succinic, sulfanilic, tartaric, toluenesulfonic, and valeric acids. Examples of suitable polymeric organic anions include, but are not limited to, those derived from the following polymeric acids: tannic acid, carboxymethyl cellulose.Such salts include acetate, adipate, aspartate, benzoate, besylate, bicarbonate, carbonate, bisulfate, sulfate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfonate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate, hydrogen phosphate, dihydrogen phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, and xinofoate.

[0437] For example, if the compound is anionic, or has a functional group which can be anionic (e.g. -RCOOH can be -RCOO ), then salts of cations or bases can be formed. Salts of the compounds of the application include those derived from inorganic bases, such as ammonium, potassium, sodium and lithium. Salts of the compounds of the application also include those derived from organic bases, such as organic amines, including primary, secondary and tertiary amines. Other salts include those derived from magnesium, calcium, barium, zinc, aluminum and copper. -), then base salts can be formed with suitable cations. Examples of suitable inorganic cations include, but are not limited to, metal cations such as alkali or alkaline earth cations, ammonium and substituted ammonium cations, and amines. Examples of suitable metal cations include sodium (Na + ), potassium (K + ), magnesium (Mg 2+ ), calcium (Ca 2+ ), zinc (Zn 2+ ), and aluminum (Al 3+ ). Examples of suitable organic cations include, but are not limited to, ammonium ions (i.e., NH4 + ) and substituted ammonium ions (e.g., NH3R + , NH2R2 + , NHR3 + , NR4 + ). Examples of some suitable substituted ammonium ions are those derived from ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, tromethamine, and amino acids such as lysine and arginine. An example of a common quaternary ammonium ion is N(CH3)4 +Examples of suitable amines include arginine, N,N'-dibenzylethylene-diamine, chloroprocaine, choline, diethylamine, diethanolamine, dicyclohexylamine, ethylenediamine, glycine, lysine, N-methylglucamine, olamine, 2-amino-2-hydroxymethyl-propane-1,3-diol, and procaine. A discussion of useful acid addition and base salts is found in S. M. Berge et al., J. Pharm. Sci. (1977) 66:1-19; see also Stahl and Wermuth, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (2011).

[0438] Pharmaceutically acceptable salts can be prepared using various methods. For example, a compound of the disclosure can be reacted with a suitable acid or base to give the desired salt. A precursor of a compound of the disclosure can also be reacted with an acid or base to remove an acid or base-labile protecting group or to open a lactone or lactam group of the precursor. Additionally, a salt of a compound of the disclosure can be converted to another salt by treatment with a suitable acid or base, or by contact with an ion exchange resin. After the reaction, if the salt precipitates from solution, it can be isolated by filtration, or it can be recovered by evaporation. The degree of ionization of the salt can vary from completely ionized to almost non-ionized.

[0439] It can be convenient or desirable to prepare, purify, and / or handle a corresponding solvate of the active compound. The term “solvate” describes a molecular complex comprising the compound and one or more pharmaceutically acceptable solvent molecules (e.g., EtOH). The term “hydrate” is a solvate where the solvent is water. Pharmaceutically acceptable solvates include those in which the solvent can be isotopically substituted (e.g., D2O, acetone-d6, DMSO-d6).

[0440] The currently accepted classification system for solvates and hydrates of organic compounds distinguishes between solvates and hydrates with separation sites, channels, and metal ion coordination. See, for example, K.R. Morris (H.G. Rittain ed.) Polymorphism in Pharmaceutical Solids (1995). Separation site solvates and hydrates are those in which solvent (e.g., water) molecules are separated from each other by the intervention of organic compounds and thus do not come into direct contact. In channel solvates, solvent molecules reside in lattice channels, where they are adjacent to other solvent molecules. In metal ion coordinated solvates, solvent molecules are bonded to metal ions.

[0441] When solvent or water is tightly bound, the complex will have a well-defined stoichiometry independent of humidity. However, when solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water or solvent content will depend on humidity and drying conditions. In this case, non-stoichiometry is often observed.

[0442] Synthesis strategy

[0443] The methods of this invention may include one or more of the techniques described below. Details of common reactions, including oxidation, reduction, etc., separation techniques (extraction, evaporation, precipitation, chromatography, filtration, grinding, crystallization, etc.), and analytical procedures, may be omitted in some schemes and embodiments, as are known to those skilled in the art of organic chemistry. Details of such reactions and techniques can be found in numerous publications, including Richard Larock, Comprehensive Organic Transformations, A Guide to Functional Group Preparations, 2nd Ed (2010), and the multi-volumeseries edited by Michael B. Smith and others, Compendium of Organic Synthetic Methods (1974 et seq.). Starting materials and reagents may be obtained from commercial sources or prepared using literature methods. Some reaction schemes may omit minor products arising from chemical transformations (e.g., alcohols from the hydrolysis of esters, CO2 from the decarboxylation of diacids, etc.). Furthermore, in some cases, reaction intermediates may be used in subsequent steps without separation or purification (i.e., in situ).

[0444] In some of the reaction schemes and examples below, certain compounds can be prepared using protecting groups, which prevent undesirable chemical reactions from occurring at other reactive sites. Protecting groups can also be used to enhance the solubility of a compound or otherwise modify the physical properties of a compound. For a discussion of protecting group strategies, descriptions of materials and methods for installing and removing protecting groups, and a compilation of useful protecting groups for common functional groups, including amines, carboxylic acids, alcohols, ketones, aldehydes, and the like, see T. W. Greene and P. G. Wuts, Protecting Groups in Organic Chemistry, 4th Edition, (2006) and P. Kocienski, Protective Groups, 3rd Edition (2005). th

[0445] Generally, the chemical transformations described throughout the specification can be carried out using amounts of reactants that are essentially stoichiometric, although certain reactions can benefit from the use of an excess of one or more reactants. Additionally, many of the reactions disclosed throughout the specification can be carried out at about room temperature (RT) and ambient pressure, although depending on reaction kinetics, yield, etc., some reactions can be run at elevated pressure or with higher temperatures (e.g., reflux conditions) or lower temperatures (e.g., -78 °C to 0 °C). Any reference in the present disclosure to stoichiometric ranges, temperature ranges, pH ranges, etc., whether or not the word “range” is explicitly used, also includes the indicated endpoints.

[0446] ​Many chemical transformations can also employ one or more compatible solvents, which can influence the rate and yield of the reaction. Depending on the nature of the reactants, one or more solvents can be polar protic solvents (including water), polar aprotic solvents, non-polar solvents, or some combination. Representative 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., dichloromethane, chloroform, carbon tetrachloride); aliphatic alcohols (e.g., methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-methyl-1-propanol, 2-butanol, 2-methyl-2-propanol, 1-pentanol, 3-methyl-1-butanol, 1-hexanol, 2-methoxyethanol, 2-ethoxyethanol, 2-butoxyethanol, 2-(2-methoxyethoxy)ethanol, 2-(2-ethoxyethoxy)ethanol, 2-(2-butoxyethoxy)ethanol); ethers (e.g., diethyl ether, diisopropyl 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 (e.g., methyl acetate, ethyl acetate); nitrogen-containing solvents (e.g.: formamide, N,N-dimethylformamide, acetonitrile, N-methylpyrrolidinone, pyridine, quinoline, nitrobenzene); sulfur-containing solvents (e.g., carbon disulfide, dimethyl sulfoxide, tetrahydrothiophene-1,1-dioxide); and phosphorus-containing solvents (e.g., HMPA, hexamethylphosphoramide).

[0447] Further particular and preferred aspects are set out in the accompanying independent and dependent claims. Features of the dependent claims can be suitably combined with features of the independent claims and with those other than explicitly set out in the claims. BRIEF DESCRIPTION OF DRAWINGS

[0448] Embodiments of the application will now be further described with reference to the drawings in which:

[0449] Figure 1 is a plot of drug serum concentration (ng / mL) in mouse plasma samples post SnMP (38 mg / kg, PO) administration analyzed by LC-MS / MS. Mean ± SD values were plotted against time in a line graph (semi-log).

[0450] Figure 2 is a plot of drug serum concentration (ng / mL) in mouse plasma samples post KCL-HO-1 i(NS) (44 mg / kg, PO) administration analyzed by LC-MS / MS. Mean ± SD values were plotted against time in a line graph (semi-log).

[0451] Figure 3 is Figure 1 and Figure 2 a combination chart of data in

[0452] Figure 4 A and Figure 4 B shows a schematic of the i.p. dosing strategy of KCL-HO-1 i or SnMP and / or Fluorouracil (5-FU) or Gemcitabine or vehicle in MMTV-PyMT mice carrying established tumors.

[0453] Figure 4 C to Figure 4 G shows growth curves of established spontaneous tumors in MMTV-PyMT mice that have received single treatment of vehicle, SnMP, KCL-HO-1 i, 5-FU or Gemcitabine according to the dosing strategies shown in Figure 41 and Figure 4 B. Each line represents an individual tumor and mouse.

[0454] Figure 4 H to Figure 4 J shows growth curves of established spontaneous tumors in MMTV-PyMT mice that have received combination treatment of SnMP / 5-FU, KCL-HO-1 i / 5-FU and KCL-HO-1 i / Gemcitabine according to the dosing strategies shown in Figure 4 A and Figure 4 B. Each line represents an individual tumor and mouse.

[0455] Figure 4 K shows the mean of the growth curves of mice with established spontaneous tumors given vehicle, KCL-HO-1 i, 5-FU, or KCL-HO-1 i and 5-FU combination as indicated.

[0456] Figure 4 L shows the mean of the growth curves of mice with established spontaneous tumors given vehicle, KCL-HO-1 i, Gemcitabine, or KCL-HO-1 i and Gemcitabine combination as indicated.

[0457] Figure 4 shows tumor size on day 8 after start of treatment in MMTV-PyMT mice that have received SnMP, KCL-HO-1 i in combination with Fluorouracil (5-FU). Each dot represents an individual mouse. *P < 0.05.

[0458] Figure 5A shows growth curves of established spontaneous tumors in MMTV-PyMT mice that have received commercial SnMP, SnMP synthesized according to the application (Vegan SnMP) and / or fluorouracil (5-FU) as indicated. Each line represents an individual mouse.

[0459] Figure 6 A shows a schematic of the experiments and strategy used to determine plasma concentrations.

[0460] Figure 7 B shows a graph of plasma concentrations over time in male mice that received either intraperitoneal (left panel) or oral (right panel) administration of commercial SnMP, SnMP synthesized according to the application (veSnMP) or KCL-HO1 i.

[0461] Figure 7 C shows HO-1 luc / eGFP A schematic of the mouse model (upper panel) and experimental flowchart (lower panel) is reported.

[0462] Figure 7 D shows HO-1 luc / eGFP Representative bioluminescence images of vehicle, commercial SnMP, SnMP synthesized according to the application (VeSnMP) or KCL-HO1 i in the whole body of reporter mice (C) at T0 (upper panel) and after 24h treatment (lower panel) are shown.

[0463] Figure 7 E shows HO-1 luc / eGFP Relative quantification (photons / sec) of luciferase expression in whole body images as shown in (D) in reporter mice (C) is shown.

[0464] Figure 7 F shows HO-1 luc / eGFP Changes in HO-1 expression normalized to vehicle treated group in individual mouse tissues (from those shown in D) in reporter mice (C) are shown.

[0465] Figure 7 G shows the evaluation of the inhibition of HO-1 activity in rat microsomes using a range of doses of KCL-HO-1 i and SnMP.

[0466] Figure 7 A shows a schematic of the dosing strategy of KCL-HO-1 i and gemcitabine in MMTV-PyMT mice.

[0467] Figure 8 B shows the tumor volume (mm3) in each mouse. 3) over time (days). KCL-HO-1i was delivered by oral delivery, and gemcitabine was delivered by intraperitoneal administration. Dotted line indicates start of treatment.

[0468] Figure 8 A bar graph showing CD8 T cell infiltration into tumors of MMTV-PyMT mice treated with vehicle, commercial SnMP (cSnMP), or KCL-HO-1i.

[0469] Figure 9 A shows a schematic representation of the intraperitoneal (i.p.) dosing strategy of non-immune IgG and immune depleting anti-CD8a antibody administered in combination with KCL-HO-1i and gemcitabine in MMTV-PyMT mice.

[0470] Figure 10 B shows growth curves of each individual mouse treated with non-immune IgG.

[0471] Figure 10 C shows growth curves of each individual mouse treated with immune depleting anti-CD8a antibody.

[0472] Figure 10 D shows growth curves of each individual mouse treated with immune depleting anti-CD8a antibody, KCL-HO-1i, and gemcitabine.

[0473] Figure 10 E shows growth curves shown as line graphs, where each line shows the mean value for each treatment.

[0474] Figure 10 A shows a schematic representation of the intraperitoneal (i.p.) dosing strategy of KCL-HO-1i and / or 5-FU or gemcitabine or vehicle in MMTV-PyMT mice carrying established tumors. Tumors were analyzed by flow cytometry 36 hours after the start of treatment.

[0475] Figure 11 B shows changes in tumor growth during the 36 hours of treatment.

[0476] Figure 11 C shows stromal composition of enzymatically dispersed tumors for each of the different treatments.

[0477] Figure 11 D shows a bar graph representing CD8 + T cell infiltration in the tumor microenvironment for each of the different treatments.

[0478] Figure 11 E shows a bar graph representing CD8 +A bar chart of T cell effector function, which is assessed by their IFN-γ expression levels.

[0479] Figure 11 F shows the CD8 matrix representing the solvent and SnMP. + A bar chart showing the infiltration of T cells in the tumor microenvironment.

[0480] Figure 11 G shows the live (7AAD) through FACS gate. - CD45 + CD3 + CD8 + A representative scatter plot of T cells, showing the expression of CD44 and CD62L in solvent-treated mice.

[0481] Figure 11 H shows the different treatment groups (e.g.) Figure 11 (G indicates CD8) + Histogram of T cell subsets.

[0482] Figure 11 A schematic diagram illustrates a dosing strategy for mice carrying established MMTV-PyMT tumors, in which these mice are treated with KCL-HO-1i and / or 5-FU or gemcitabine or solvent, and the tumor tissue is analyzed by bulk RNAseq analysis after 36 hours.

[0483] Figure 12 B shows a Venn diagram of the intersection between all DEGs and their groups for the corresponding treatment relative to the solvent.

[0484] Figure 12 C shows a Venn diagram illustrating the intersection between DEG and its groups when the KCL-HO-1i-based treatment is upregulated relative to all solvents.

[0485] Figure 12 D shows a heatmap of hierarchical clustering of secretory genes (91 genes) in the DEG (419 genes) that were co-upregulated across treatment groups.

[0486] Figure 12 E shows a Venn diagram of the intersection between all upregulated DEGs and their groups, including those treated with chemotherapy.

[0487] Figure 12 F shows a Venn diagram of the intersection between chemokines and upregulated DEG and their groups associated with dual therapy treatment.

[0488] Figure 12G shows a heatmap of hierarchical clustering of DEGs of chemokines and cytokines upregulated in relation to the doublet processing group.

[0489] Figure 12 A shows a schematic representation of the dosing strategy for oral delivery of KCL-HO-1 i and / or gemcitabine or vehicle in MMTV-PyMT mice carrying established tumors.

[0490] Figure 13 B shows Figure 13 Tumor growth curves for the respective treatments shown in A.

[0491] Figure 13 C is a graph showing the body weight of the mice during the respective treatments.

[0492] Figure 13 D shows the ratio of serum liver aspartate aminotransferase (AST) to alanine aminotransferase (ALT) in the blood of the respective treatments.

[0493] Figure 13 E shows the blood immune cell composition of the respective treatments.

[0494] Figure 13 F shows a heatmap of animal body features assessed on day 21 of treatment.

[0495] Figure 13 A shows a schematic representation of the dosing strategy for oral delivery of KCL-HO-1 i and / or i.p. delivery of gemcitabine or vehicle in C57B1 / 6 mice carrying established subcutaneous sarcoma tumors (MN-MCA-1 cell line).

[0496] Figure 14 B shows Figure 14 Tumor growth curves for the respective treatments shown in A.

[0497] Figure 14 A shows the tumor volume of mice receiving either KCL-HO-1 i (blue curve) or vehicle (black curve) daily from day 12 after tumor injection for 14 days.

[0498] Figure 15 B shows a histogram representing the counted metastatic nodules in the lungs harvested from day 26 after tumor injection. DETAILED DESCRIPTION

[0499] EMBODIMENT

[0500] Surprisingly, the inventors have discovered that KCL-HO-1i shows unexpected anticancer activity in in vivo studies and proves superior to the commercial SnMP dichloride (see section 2). KCL-HO-1i appears to have an excellent solubility profile compared to the commercially available SnMP.

[0501] KCL-HO-1i is synthesized from SnMP as shown below. Typically, SnMP can be contacted with a source of phosphate at a predetermined temperature for a predetermined time, or MP can be contacted with a source of tin and a source of phosphate at a predetermined temperature for a predetermined time.

[0502] 1) Synthesized KCL-HO-1i

[0503]

[0504] Scheme 2. Synthesis of KCL-HO-1i using tin pyrophosphate.

[0505] 2) KCL-HO-1i as a HO inhibitor with antitumor efficacy.

[0506] We have explored KCL-HO-1i preclinically and demonstrated its superiority over the dichloride SnMP (as shown in Figure 15 , Figure 3 , Figure 4 , Figure 5 and Figure 7 ). Administration of KCL-HO-1i to MMTV-PyMT tumor-bearing mice has demonstrated that KCL-HO-1i in combination with chemotherapy 5-FU shows superior antitumor efficacy ( Figure 9 ), with KCL-HO-1i having surprisingly eliminated all tumor burden at day 8 after the start of treatment while SnMP treated animals still had residual disease ( Figure 4 ). Thus, KCL-HO-1i in combination with 5-FU provides a measurable improvement in the antitumor immune response compared to the standard commercial SnMP ( Figure 5 ).

[0507] Currently, MP is synthesized using animal product starting materials, in particular chlorinated hematin.

[0508] There is a need for a synthesis approach to improve manufacturing efficiency and scale, and to address ethical and religious concerns, and which is vegetarian friendly as it does not contain animal products. Surprisingly, the inventors provide a method to synthesize MP using a method that allows for large scale manufacturing of MP without the use of animal products.

[0509] 3) Large scale manufacturing of synthesized KCL-HO-1i.

[0510] The synthetic route shown in Scheme 3 below is a possible route to synthesize KCL-HO-1i without the need for animal starting materials.

[0511] SnMP prepared using the synthetic route according to the method of the present application is active in delivering anti-tumor efficacy in combination with chemotherapy 5-FU in the MMTV-PyMT mouse cancer model (see Figure 5 ).

[0512]

[0513]

[0514] Scheme 3. Optimized synthesis for large scale preparation of the synthesized tin mesoporphyrin phosphate salt (KCL-HO-1i) according to the present application.

[0515] KCL-HO-1i can be synthesized using the MP produced in Stage 4 or SnMP prepared in Stage 5 by contacting the MP or SnMP with a source of tin and a source of phosphate, for example tin pyrophosphate as described above in 1.

[0516] Stage 1

[0517]

[0518] Pyrrole A (1) (200 g), pyrrole B (2) (309 g) and methanol (3.99 L) were charged into a reactor under nitrogen and the contents heated to reflux. A solution of p-toluenesulfonic acid monohydrate (1.21 g) in methanol (10 mL) was added over 3 minutes. The reaction mass was stirred at reflux for 32 minutes then cooled to 0 °C over 2:30 hours at a controlled rate. Spontaneous crystallization occurred between 38 and 36 °C. Once at 2 °C, the slurry was discharged to a filter, deliquored and the vessel washed to the filter using methanol (2 x 500 mL) cooled to <5 °C. The filter cake was fully deliquored and dug out to a drying tray, vacuum dried at 40 °C for 8 hours to give the product (3) (286.8 g, 90.7% yield).

[0519] Figure 6

[0520]

[0521] Compound (3) (210.0 g), tetrahydrofuran (1050 mL) and triethylamine (147 mL) were charged into a flask and the contents stirred until a solution was formed. 5% Palladium on alumina (42.0 g) was charged into the flask and hydrogen was bubbled through at 500 mL min -1Hydrogen was sprayed onto the mixture while stirring for 52 minutes, during which time the temperature of the contents of the flask increased from 17°C to 37°C. After this time, the reaction temperature began to decrease, and the gas flow rate emanating from the batch was visually observed to increase significantly. After a reaction time of 60 minutes, the mixture was passed through a 25 mm deep diatomaceous earth bed into a 3 L pear-shaped flask. The filter bed was washed with tetrahydrofuran (4 × 100 mL). The filtrate and washings were combined and concentrated under reduced pressure to give a moist, off-white solid, which was then vacuum dried at 40°C for 20 hours to give a dried intermediate (174.2 g, yield 98.6%), which was ready for use without purification (via...). 1 H NMR confirmed identity.

[0522] Phase 2

[0523]

[0524] A flask containing p-toluenesulfonic acid monohydrate (372.4 g), chloroform (10.61 L), and methanol (2.12 L) was placed under a nitrogen atmosphere. A solution of crude compound (26) TEA salt (174.1 g) from stage 2 and dialdehyde (157.6 g) in 5:1 chloroform / methanol (1.50 L) was added over 16:30 hours using an HPLC pump. 3 × 40 mL of 5:1 chloroform / methanol was added as a flushing solution for the container and tubing. 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 jetted through the mixture for 6 hours. The mixture was then stirred with the mixture exposed to atmosphere for approximately 10 hours. The mixture was washed with deionized water (2 × 2.50 L), followed by a 20% w / w potassium carbonate aqueous solution (2.10 L, 2.5 kg) and an additional 2.50 L of water. The chlorinated solution was concentrated under reduced pressure to obtain a dark brown / purple oily substance, which was then vacuum dried at 40°C for 20 hours to obtain a dried intermediate (21) (255.6 g, yield 99%), which can be used without further purification or analysis.

[0525] A solution of crude zinc porphyrin complex (21) (511.7 g) and 5% v / v sulfuric acid in methanol (2.50 L) was charged to a flask and stirred until the solids dissolved. Stirring was continued for 4 hours, after which time the mixture was transferred to a 20 litre vessel and diluted using dichloromethane (5.00 L). The mixture was washed using deionised water (2.50 L). The aqueous layer was back-extracted using dichloromethane (2 x 0.5 L). The chlorinated phases were combined and washed using deionised water (2.50 L), then using 20% w / w aqueous potassium carbonate solution (2.10 L, 2.5 kg) and further water (2.50 L). The chlorinated solution was concentrated under reduced pressure to give a dark brown / black oily material which was dried under vacuum at 40 °C for 20 hours to give dry stage 3 crude product (6) (490.1 g).

[0526]

[0527] Stage 3 crude product (480 g) was dissolved in dichloromethane (ca. 3.5 L) and loaded onto a 5 kg HP-Sphere silica column in a Biotage Flash 150 L system which had been equilibrated using dichloromethane. Elution was carried out using dichloromethane (ca. 800 mL min -1 The column was eluted using dichloromethane (40 L), then using 0.5% methanol in dichloromethane (130 L) to take ca. 10 L fractions. Elution was continued using 0.5% methanol in dichloromethane (30 L) to take a further 6 x ca. 5 L fractions. The desired fractions were combined and concentrated under reduced pressure to give stage 3 product (214.3 g, 44.3% yield) as a brown / purple solid.

[0528] Phase 3

[0529]

[0530] Compound (6) from stage 3 (85.0 g), tetrabutylammonium chloride (850 mg), dichloromethane (1.70 L), methanol (1.70 L) and 2M aqueous sodium hydroxide solution (268 mL) were charged to a flask and the contents heated to reflux (45 °C). The mixture was stirred at reflux for 4 hours, after which time the reaction was complete 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. The mixture was stirred for 1 hour, then drained onto a filter, drained and the vessel washed through to the filter using a 1 : 1 mixture of dichloromethane and methanol (2 x 150 mL), then water (2 x 100 mL). The filter cake was fully drained and dug out.

[0531] A wet product filter cake, deionized water (300 mL) and methanol (300 mL) were charged to a flask and the contents were heated to 70 °C and stirred for 30 minutes, then cooled to room temperature. The slurry was drained to a filter, deliquored, and the vessel washed to the filter using a 1 : 1 mixture of methanol and deionized water (2 x 150 mL), followed by methanol (100 mL). The filter cake was fully deliquored and scooped into a drying tray, vacuum dried at 45 °C for 20 hours to yield Stage 4 product (71.6 g) as a brown / purple powder. The liquids from both filtrations were concentrated under reduced pressure to yield a slurry of approximately 600 mL which was drained to a filter and deliquored. The filter cake was washed with deionized water (2 x 100 mL). The filter cake was fully deliquored and scooped into a drying tray, vacuum dried at 45 °C for 20 hours to yield a second harvest of Stage 4 product (7) (7.2 g) as a brown / purple powder. Stage 4 products (7) were combined to yield a brown / purple powder (78.8 g, 97.3% yield).

[0532] Phase 4

[0533]

[0534] To a 50 mL round bottom flask, covered with tin foil, was added 200 mg mesoporphyrin IX (0.35 mmol, 1 eq.) and 576 mg tin (II) pyrophosphate (1.4 mmol, 4 eq.). 5 mL glacial acetic acid was added and the flask was flushed with nitrogen. The mixture was then stirred at reflux at 115 °C. Once temperature was stable, the reaction was allowed to open to air to introduce oxygen into the system, then the reaction was left for 24 hours. The mixture was then cooled to room temperature, quenched with 4 mL HPLC grade water, filtered, and air dried for 10 minutes. The filtered solid was placed in another 50 mL flask containing 3 mL HPLC grade water and 0.48 mL concentrated HC1, then stirred at 90 °C for 30 minutes. The mixture was cooled to room temperature, then filtered and washed with cold water. After air drying for 30 minutes, 125 mg of product (KCL-HO-1i) was obtained as a red powder (40% isolated yield).1H NMR (400 MHz, DMSO-d6) δ 10.36 (s, 1H), 10.13 (s, 1H), 10.09 (d, J = 4 Hz, 2H), 7.11 (s, 4H), 3.82 - 3.96 (m, 5H), 3.62 (q,, J = 14 Hz, 4H), 3.17 (d, J = 12 Hz, 12H), 2.64 (t, J = 6 Hz, 5H), 1.82 (t, J = 2 Hz, 16H), 1.25 (t, J = 6 Hz, 8H).

[0535] Alternative Stage 5

[0536] This general reaction scheme can also be applied to the preparation of SnMP by using the following alternative Stage 5.

[0537]

[0538] Tin (II) oxide (101.7 g) and acetic acid (2.50 L) were charged to a flask and the contents heated to 65 °C. A solution of Stage 4 (7) (115 g) in a mixture of formic acid (590 mL) and concentrated hydrochloric acid (43.2 g) was added dropwise over 3:40 hours while maintaining the temperature of the mixture in the flask between 60 and 70 °C. Formic acid (3 x 20 mL) was charged as a line flush; the formation of a deep red precipitate became evident after about 2:45 hours (by about 75% addition). The mixture was stirred at 63 to 68 °C for 17:25 hours. Deionized water (1.05 L) was added over 20 minutes. The mixture was allowed to cool to 20 °C over 3 hours and stirred for an additional 2 hours. The slurry was discharged to a filter, drained, and the vessel to the filter washed with deionized water (2 x 100 mL). The filter cake was drained, dug out, and recharged to a flask. 1 M aqueous hydrochloric acid (1.75 L) was charged to the flask and the contents heated to 85 °C. The mixture was stirred at 85 to 90 °C for 80 minutes, then the slurry was discharged to a filter, drained, and the vessel to the filter washed with water (2 x 250 mL). The filter cake was completely drained and dug out to a drying tray and vacuum dried at 40 °C for 40 hours to give tin mesoporphyrin (135.7 g, 88.7% yield) as a deep red powder.

[0539] An inductively coupled plasma mass spectrometry (ICP-MS) comparison was performed between two samples of SnMP (tin chloride salt of mesoporphyrin IX) and KCL-HO-1i to provide elemental analysis of the tin content of these compounds.

[0540] Phase 5 Sample Powder in solute (g / L) Sn (pg / g) 3.49 340508.46 SnMP 2.94 949046.39 KCL-HO-1 i (1) 3.88 1081682.85

[0541] Thus, this elemental analysis data shows that the average of Sn for KCL-HO1i is 3 times more than SnMP.

[0542] This general reaction scheme can also be applied to the synthesis of the tartrate salt of tin mesoporphyrin IX (TA-191-149) by using the following alternative Stage 5.

[0543]

[0544] To a 50 mL round bottom flask, covered with tin foil, was added 200 mg mesoporphyrin IX (0.35 mmol, 1 eq.) and 372.5 mg tin(II) tartrate (1.4 mmol, 4 eq.). 5 mL glacial acetic acid was added and the flask was flushed twice with nitrogen. The mixture was then stirred under reflux at 120 °C. Once the temperature was stable, the reaction was allowed to open to air to introduce oxygen into the system and the reaction was left for 24 hours. The mixture was then cooled to room temperature, quenched with 4 mL HPLC grade water, filtered and air dried for 10 minutes. The filtered solid was placed in another 50 mL flask containing 3.5 mL 1 M HC1 and then stirred at 90 °C for 30 minutes. The mixture was cooled to room temperature and then filtered and washed with cold water. After air drying for 30 minutes, 125 mg of product was obtained as a brick red powder (32% isolated yield). M.P: 287.2 °C; 1 H NMR (400 MHz, Methanol-d4) d 11.21 (br. s., 1H), 11.01 (br. s., 1H), 10.97 (br. s., 2H), 4.45 (d, J = 7.06 Hz, 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).

[0545] This general reaction scheme can also be applied to the synthesis of the oxalate salt of tin mesoporphyrin IX (TA-191-151) by using the following alternative Stage 5.

[0546]

[0547] To a 50 mL round bottom flask, covered with tin foil, was added 200 mg mesoporphyrin IX (0.35 mmol, 1 eq.) and 372.5 mg tin(II) tartrate (1.4 mmol, 4 eq.). 5 mL glacial acetic acid was added and the flask was flushed twice with nitrogen. The mixture was then stirred under reflux at 120 °C. Once the temperature was stable, the reaction was allowed to open to air to introduce oxygen into the system and the reaction was left for 24 hours. The mixture was then cooled to room temperature, quenched with 4 mL HPLC grade water, filtered and air dried for 10 minutes. The filtered solid was placed in another 50 mL flask containing 3.5 mL 1 M HC1 and then stirred at 90 °C for 30 minutes. The mixture was cooled to room temperature and then filtered and washed with cold water. After air drying for 30 minutes, 125 mg of product was obtained as a brick red powder (32% isolated yield). M.P: 287.2 °C; 1H NMR (400 MHz, Methanol-d4) d 11.08 (d, J = 8.34 Hz, 1H), 10.89 (d, J = 6.69 Hz, 1H), 10.85 (d, J = 5.41 Hz, 2H), 4.19 - 4.39 (m, 5H), 3.67 - 3.93 (m, 14H), 3.27 - 3.47 (m, 5H), 3.23 (s, 1H), 1.91 (q, J = 7.40 Hz, 7H).

[0548] This general reaction scheme can also be applied to the synthesis of mesoporphyrin IX methanesulfonate (TA-191-153) by using the following alternative Stage 5.

[0549]

[0550] To a 50 mL round bottom flask covered with tin foil was added 200 mg mesoporphyrin IX (0.35 mmol, 1 eq.) and 290 mg tin (II) methanesulfonate (1.4 mmol, 4 eq.). 5 mL glacial acetic acid was added and the flask was flushed twice with nitrogen. The mixture was then stirred at reflux at 120 °C. Once the temperature was stable, the reaction was allowed to open to air to introduce oxygen into the system and then the reaction was left for 24 hours. The mixture was then cooled to room temperature, quenched with 4 mL HPLC grade water, filtered and air dried for 10 minutes. The filtered solid was placed in another 50 mL flask containing 3.5 mL 1 M HC1 and then stirred at 90 °C for 30 minutes. The mixture was cooled to room temperature and then filtered and washed with cold water. After air drying for 30 minutes, 72 mg of product was obtained as a red powder (24% isolated yield). M.P: 257.8 °C; 1 H NMR (400 MHz, Methanol-d4) d 11.25 (d, J = 16.05 Hz, 1H), 10.94 - 11.14 (m, 3H), 4.71 - 4.80 (m, 4H), 4.39 - 4.53 (m, 4H), 3.91 - 4.04 (m, 12H), 3.47 (t, J = 7.24 Hz, 4H), 2.70 (s, 6H), 2.03 (t, J = 7.57 Hz, 6H).

[0551] 4) Synthesis of pyrrole A

[0552]

[0553] The selective reduction step (iii) can be achieved by: (a) Wolff-Kishner reduction using N2H2, KOH; (b) Clemmensen reduction using Zn-Hg, HC1; (c) catalytic hydrogenation using Pt-C, H2 under pressure; or (d) using TsNHNH2 followed by NaBH3CN.

[0554] 4) Synthesis of pyrrole B

[0555]

[0556] (a) KOH, benzyltrimethylammonium bromide, CHCl3; (b) pent-3-yn-2-one, 5 mol% Cu catalyst (as Cu2O), 20 mol% PPh3, dioxane, 100 °C for 6-8 hours, see (19); (c) selective reduction using any of the following: (i) Wolff-Kishner reduction using N2H2, KOH; (ii) Clemmensen reduction using Zn-Hg, HC1; (iii) catalytic hydrogenation using Pt-C, H2 under pressure; or (iv) using TsNHNH2 followed by NaBH3CN.

[0557] 5) Synthesis of dialdehyde

[0558]

[0559] Further details regarding step (v) are found in (20).

[0560] KCL-HO-1 i (2)

[0561] Pharmacokinetic (PK) studies were performed in male mice ICR after oral (PO) administration of SnMP(CR) and KCL-HO-1 i(NS) at 38 mg / kg and 44 mg / kg, respectively. Plasma samples were collected at predetermined times after PO administration. Two formulations containing SnMP(CR) and KCL-HO-1 i7(NS) were prepared as shown in Table 1 below. The dosing volume for SnMP(CR) was 9.5 mL / kg and for KCL-HO-1 i(NS) was 11 mL / kg.

[0562] SnMP(CR) formulation (4 mg / mL). 23.38 mg SNMP(CR) powder in 1.17 mL 0.1 M sodium hydroxide (NaOH); once completely dissolved, 4.68 mL 0.5 M sodium bicarbonate (NaHCO3) was added at pH 7 to reach a final concentration of 4 mg / mL.

[0563] KCL-HO-1 i(NS) formulation (4 mg / mL). 26.68 mg KCL-HO-1 i(NS) powder in 1.334 mL 0.1 M NaOH; once completely dissolved, 5.336 ml 0.5 M NaHC03 was added at pH 7 to reach a final concentration of 4 mg / mL.

[0564] Male ICR mice of 25 ± 5 g were obtained from BioLasco, Taiwan, China (under license from Charles River Laboratories). The animals were acclimated for 3 days prior to use and confirmed to be in good health. All animals were maintained in a sanitary environment with controlled temperature (20 to 24 °C), humidity (30% to 70%), and a 12-hour light / dark cycle. Sterile standard laboratory diet [MFG (Oriental Yeast Co., Ltd., Japan)] and autoclaved tap water were available ad libitum. All aspects of this work, including animal housing, experiments, and handling, were generally conducted in our AAALAC-accredited facility in accordance with the Guide for the Care and Use of Laboratory Animals: Eighth Edition (National Academy Press, Washington, D.C., 2011). Animal care and use protocols were reviewed and approved by the IACUC of Pharmacology Discovery Services Taiwan, China, Ltd.

[0565] The animal dosing design is shown in Table 2 below.

[0566] Pharmacokinetic (PK) study

[0567] Blood aliquots were collected from euthanized mice by cardiac puncture (approximately 300 μL), placed in heparin lithium-coated tubes, mixed gently, and centrifuged at 2,500 x g for 15 minutes at 4 °C within 1 hour of collection. Plasma samples were then harvested and kept frozen at -70 °C until further processing.

[0568] Mouse plasma sample collection (parallel)

[0569] Plasma samples were processed using methanol precipitation and analyzed by LC-MS / MS. The exposure levels (ng / mL) of the formulations [SnMP or KCL-HO-1i] in plasma samples were then determined by LC-MS / MS. The exposure levels (ng / mL) of SnMP in mouse plasma samples after SnMP (38 mg / kg, PO) administration are shown in Quantitative bioanalysis (plasma samples) and pharmacokinetics Figure 1A, KCL-HO-1i as shown in Figure 1 Figure IB, and the combination plot as shown in Figure 2 Figure 1C.

[0570]

[0571]

[0572] (a) This is based on visual observation: S: soluble; SS: slightly soluble; I: insoluble (suspension or precipitate).

[0573] (b) Y: formulation was stored in brown test tubes or vials, or covered with aluminum foil.

[0574] (c) RT: freshly prepared and stored between 20 to 25 °C. 4 °C: freshly prepared and stored in the refrigerator or on ice.

[0575] Table 1: Formulations

[0576]

[0577] Table 2: Animal dosing design

[0578] In vivo anti-tumor effects of various single and combination treatments

[0579] Mice bearing established tumors (MMTV-PyM), as a model of an aggressive breast cancer, received an intraperitoneal (i.p.) dosing strategy, providing single treatments of KCL-HO-1i (25 μΜοΙ / kg / day) or SnMP (25 μΜοΙ / kg / day) or 5-FU (40 mg / kg / 4 days) or gemcitabine (64 mg / kg / 7 days) or vehicle injection, or double treatments of (i) KCL-HO-1i or SnMP in combination with (ii) 5-FU or gemcitabine (see Figure 3 A and Figure 4 B). Treatments were initiated on day 0, marked by a vertical dashed line in each of Figure 4 C through Figure 4 L. Tumor growth was monitored and the tumor volume of each individual mouse was calculated and the results are shown for the different single treatments (see Figure 4 C through Figure 4 G) and combination treatments (see Figure 4 H throughFigure 4 Growth curves for KCL-HO-1 i. KCL-HO-1 i in combination with chemotherapy (either with 5-FU or with gemcitabine) provided durable anti-tumor efficacy, as shown in Figure 4 K and Figure 4 L, where each line shows the mean (n=5 per treatment group) (**P<0.01). Surprisingly, treatment with KCL-HO-1 i and 5-FU eliminated all tumor burden by day 8 after treatment initiation, providing a measurable improvement in anti-tumor immune response relative to treatment with standard commercial SnMP and 5-FU, where animals still had residual disease (see Figure 4 ). Figure 5

[0580] Plasma concentrations were measured over the course of three phases, including (1) injection (ip) or oral gavage of SnMP (25 μMol / kg) or KCL-HO1 i (25 μMol / kg); (2) blood collection; and (3) LC / MS / MS measurement. Bioavailability (A) provides a schematic of the procedure. Male mice received either intraperitoneal administration of commercial SnMP or KCL-HO1 i (see Figure 7 (B) (left panel)) or oral administration of commercial SnMP or KCL-HO1 i (see Figure 7 (B) (right panel)). Blood aliquots were collected by cardiac puncture at 0.167, 0.5, 1, 2, 4, 6, 8, and 24 hours after oral administration of SnMP, placed in heparin lithium-coated tubes. After centrifugation, plasma samples were collected and kept at -70°C until processing. Exposure levels (ng / mL) in plasma samples for intraperitoneal administration of SnMP (blue curve), veSnMP, and KCL-HO1 i (red curve) are shown in Figure 7 (B) (left panel), and for oral administration in Figure 7 (B) (right panel). These exposure levels were determined by liquid chromatography-tandem mass spectrometry (LC-MS / MS) Figure 7 A). Plasma concentrations of SnMP and KCL-HO1 i in mouse plasma were plotted against time as mean ± SEM (standard error of the mean). The bar represents the mean, and the dots represent individual data points from individual mice.

[0581] A bioavailability study was implemented to test KCL-HO1 i against commercial SnMP using bioluminescence. Figure 7 (C) shows the HO-1 luc / eGFP Schematic of mouse model and subsequent bioluminescence strategy. HO-1 luc / eGFPMice were injected and imaged at TO and 24 hours post-treatment. Organs have been collected and imaged. Figure 7 (D) shows a comparison of HO-1 expression using vehicle, commercial SnMP or KCL-HO1i treatment. luc / eGFP Representative bioluminescence images of whole-body relative luciferase expression in mice at TO (top panel) and 24 hours post-treatment (bottom panel). Relative quantification of luciferase expression (photons / sec) in whole-body and across tissues as Figure 7 (E) is shown. Plots of total HO-1 and HO-1 expression changes in each individual tissue and mouse were obtained by normalizing to the vehicle group. Unexpectedly, KCL-HO-1i did not induce HO-1 expression, which was considered as an adverse side effect of commercial SnMP Figure 7 E, Figure 7 F). Figure 7 (G) shows the evaluation of the inhibition of HO-1 activity in rat microsomes using different doses of KCL-HO-1i and SnMP (n = 3 biological replicates), where the concentration of KCL-HO-1i and SnMP (mM) is shown in a logarithmic scale. (*P < 0.05, **P < 0.01, ****P < 0.001).

[0582] These results show that KCL-HO1i shows better in vivo availability than commercial SnMP. In vivo anti-tumor effect of oral administration of KCL-HO-1i in combination with gemcitabine

[0583] Mice carrying spontaneous tumors (MMTV-PyM), an aggressive breast cancer model, were given a combination of gemcitabine (by intraperitoneal administration) and KCL-HO-1i (by oral administration) as shown in Figure 8 A. Tumor measurements were initiated on day 0 (n = cohort of 2 mice). Tumor growth was monitored and tumor volume was calculated for each mouse (as shown in Figure 8 B, where each line represents an individual mouse and tumor). KCL-HO-1i in combination with gemcitabine can control tumor growth.

[0584] Investigation of CD8 T cell infiltration into tumors of MMTV-PyMT mice.

[0585] Tumors had reached 500 to 750 mm 3MMTV-PyMT spontaneous tumor mice. Mice were given an equivalent dose of commercial SnMP (cSnMP) or KCL-HO-1 i on days 0 and 1, and then tumors were harvested on day 2 (36 hours after the first injection). Tumors were subjected to enzymatic digestion to release individual cells, and the proportion of CD8+CD3+CD45+ T cells was assessed by flow cytometry. The infiltration of CD8 T cells into tumors of MMTV-PyMT mice treated with vehicle, commercial SnMP (cSnMP), or KCL-HO-1 i is shown as Figure 9 bar graphs in which points represent individual tumors and mice. ***P < 0.001.

[0586] Effect of immune-depleting anti-CD8a antibody on the in vivo anti-tumor effect of KCl-HO-1 i in combination with gemcitabine

[0587] Spontaneous tumor-bearing mice (MMTV-PyM) were treated with non-immune IgG or with immune-depleting anti-CD8a antibody. Mice were given the respective antibody treatment as a loading dose on day -2, followed by doses every 4 days. Starting on day 0, some of these mice were also given KCL-HO-1 i (25 μMol / kg / day) and gemcitabine (64 mg / kg / 7 days). Tumor growth was monitored, and the tumor volume was calculated for each mouse, and tumor growth curves are shown in Figure 10 B to Figure 10 E. For Figure 10 B, Figure 10 C and Figure 10 D, points represent individual tumors and mice. Figure 10 E is a line graph in which each line shows the mean and error bars for each treatment. In Figure 10 , the black dotted line marks the start of treatment (day 0), and the red line marks 250 mm 3 **P < 0.01. These results, particularly KCL-HO-1 i / gemcitabine / anti-CD8a antibody compared to KCL-HO-1 i / gemcitabine (see Figure 11 E), indicate that the anti-tumor effect of KCL-HO-1 i and gemcitabine is immunotherapeutic. Investigation of T cell infiltration into tumors and T cell effector function.

[0588] Spontaneous tumor-bearing mice (MMTV-PyM) were treated with KCL-HO-1 i (25 μMol / kg / day) and / or 5-FU (40 mg / kg / 4 days) or gemcitabine (64 mg / kg / 7 days) or vehicle. Tumors were analyzed before the start of treatment, and again 36 hours after the start of treatment. Tumor growth during the 36 hours of treatment for each treatment is shown in Figure 11The line graph of B is shown. Tumors from treated MMTV-PyMT were removed, collected, and studied by flow cytometry to assess the tumor microenvironment population by analyzing the stromal composition of the tumor. No significant changes were observed in the stromal population of most cells (see [link to graph]). Figure 11 C). From CD8 + T-cell analysis revealed that KCL-HO-1i induced CD8... + T cell influx into the tumor microenvironment is enhanced by combining KCL-HO1i with chemotherapy (see [link to relevant documentation]). Figure 11 (D histogram). KCL-HO-1i processing improves CD8. + The effector function of T cells, which is assessed by their IFN-γ expression levels, was enhanced by combining KCL-HO1i with chemotherapy (see [link to relevant documentation]). Figure 11 (Bar chart of E). There is evidence of a synergistic effect between KCL-HO1i and the tested chemotherapeutic agents. In contrast to KCL-HO1i, SnMP was found not to induce CD8+ using equivalent doses and regimens. + T cells infiltrate the tumor microenvironment in MMTV-PyMT mice (see [link]). Figure 11 (Bar chart of F). Flow cytometry involves the study and quantification of populations of interest using gated fluorescence activated cell sorting (FACS). Figure 11 G shows the live (7AAD) gated via FACS. - CD45 + CD3 + CD8 + A representative scatter plot of T cells, showing the expression of CD44 and CD62L in solvent-treated mice, and Figure 11 H indicates the different treatment groups (e.g., ... Figure 12 (G indicates CD8) + Histogram of T cell subsets.

[0589] The bar chart shows the mean, and the dots show individual data points from individual tumors and mice. The line chart shows the mean and SEM. *P<0.05, **P<0.01.

[0590] Research through batch RNA sequencing analysis

[0591] Mice bearing established MMTV-PyMT tumors were dosed by i.p. administration of KCL-HO-1 i (25 μMol / kg / day) and / or 5-FU (40 mg / kg) or gemcitabine (64 mg / kg) or vehicle and at 36 hours tumor tissue was removed, collected and analyzed by bulk RNA sequencing (n=5 mice and tumor cohorts) as described in Example 1. Figure 12 A). From this analysis, a Venn diagram showing all upregulated DEGs for the respective treatments relative to vehicle was plotted (see Figure 12 B) and DEGs showing KCL-HO-1 i treatment related upregulation (see Figure 12 C) and the intersection between groups were prepared. Also, a heatmap was prepared for secreted genes (91 genes) among DEGs (419 genes) that were commonly upregulated between treatment groups in treatment was hierarchically clustered with the scale from low to high relative to the maximum degree of change in tumors treated with vehicle (see Figure 12 D). Likewise, from this analysis, a Venn diagram showing all upregulated DEGs for treatments including chemotherapy (either 5-FU or gemcitabine alone or in combination with KCL-HO-1 i) relative to vehicle was plotted (see Figure 12 E). A Venn diagram showing chemokines and upregulated DEGs relative to vehicle and doublet treatment (KCL-HO-1 i / 5-FU or KCL-HO-1 i / gemcitabine) and the intersection between groups was plotted (see Figure 12 F). Also, a heatmap was prepared for chemokines and cytokines upregulated DEGs related to doublet treatment groups in treatment was hierarchically clustered with the scale from low to high relative to the maximum degree of change in tumors treated with vehicle (see Figure 13 G). KCl-HO-1 i in combination with gemcitabine oral in vivo treatment and analysis of animal body features and blood

[0592] Mice bearing established MMTV-PyMT tumors were dosed by oral administration of KCL-HO-1 i (25 μMol / kg / day) and / or i.p. delivery of gemcitabine (64 mg / kg / 7 days) or vehicle (see Figure 13 A). Figure 13 B shows a line graph of tumor growth where each line shows the mean of mice that underwent each treatment, from Figure 13 As can be seen in B, KCL-HO-1 i in combination with gemcitabine can control tumor growth. The body weight of mice was measured during treatment and it was found to remain close to the initial body weight (see Figure 13C). Blood samples were collected 21 days after treatment and plasma was separated from blood by centrifugation. Serum liver aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were assessed by enzyme-linked immunosorbent assay (ELISA) and the ratio of AST / ALT was plotted in a histogram (see Figure 13 D) (n=5 per treatment group). It was found that there were no significant changes in any of the treatment groups compared to treatment with vehicle (see Figure 13 D). Blood immune cell composition was measured (see Figure 13 E), and animals were examined for physical signs of distress by behavioral responses at day 21 and these were scored on a scale of 0 to 5 (see Figure 14 F). No signs of distress were found in the animals and these results, along with body weight measurements and blood analysis, indicated that the treatments were non-toxic at the doses tested.

[0593] Bar graphs show the mean values, and dots show individual data points from individual tumors and mice. Line graphs show the mean and SEM. *P < 0.05, **P < 0.01.

[0594] Histopathological evaluation of organs from mice treated with KCL-HO-1 i and / or gemcitabine.

[0595] Mice carrying established tumors were treated with oral gavage delivery of KCL-HO-1 i (25 μΜοΙ / kg / day, with a 5 days on, 2 days off schedule) and / or intraperitoneal (i.p.) delivery of gemcitabine (64 mg / kg / 7 days) or vehicle (n=5 per treatment group) in MMTV-PyMT mice. At 21 days after the start of treatment, the indicated tissues were harvested for histological examination and tissue sections from each mouse were cut and stained using hematoxylin and eosin staining (H&E) and scored by a pathologist. These scores used a non-linear semi-quantitative grading system from 0 to 5, where 0 = no significant changes, 5 = entire organ or tissue affected at every observation.

[0596] From the histological examination of tissues from the brain, heart, kidney, and liver, little or no significant changes were observed for all treatments. However, the pathologist scored the mice treated with KCL-HO-1 i and gemcitabine doublet up to 3 for mononuclear inflammatory cell infiltration and multifocal (perivascular) pathology in the histological examination of lung tissue. In addition, for the lung, the pathologist scored the mice treated with KCL-HO-1 i and gemcitabine doublet up to 2 for focal (alveolar) mixed inflammatory cell infiltration. However, in the histological examination of lung tissue, the lung was scored as 0 when scored for metastasis and for alveolar macrophage (alveolar) pathology.

[0597] Further histopathological evaluation of the lungs of non-tumor bearing C57B1 / 6 mice was performed in which mice were dosed with KCL-HO-1 i (25 μMol / kg / day) and gemcitabine (64 mg / kg / 7 days) doublet or vehicle by oral administration for 21 days. At the end of treatment, lungs were then excised from one cohort of mice and the second cohort of mice was not treated for 30 days prior to sacrifice. Lung sections were stained with H&E and evaluated by a pathologist and scored on the same non-linear semi-quantitative grading system. For the doublet treatment, at the end of treatment, mononuclear inflammatory cell infiltration and multifocal (perivascular) pathology were again observed in the histological examination of lung tissue scored up to 3 and focal (alveolar) mixed inflammatory cell infiltration was observed scored up to 2. However, lower scores for these pathologies were observed 30 days after the end of treatment and some mice scored 0 in the histological examination of lung tissue.

[0598] These histopathological evaluations indicate that the KCL-HO-1 i and gemcitabine doublet treatment showed some lung toxicity, but these issues started to resolve after the treatment was stopped. The absence of metastasis after the doublet treatment indicates that KCL-HO-1 i can have an anti-metastatic effect.

[0599] Further in vivo anti-tumor effect of oral administration of KCl-HO-1 i in combination with gemcitabine

[0600] Mice (C57B1 / 6) bearing established subcutaneous sarcoma tumors (MN-MCA-1 cell line) were orally dosed with KCL-HO-1 i (25 μMol / kg / day) and / or intraperitoneally administered with gemcitabine (64 mg / kg / 7 days) or vehicle as Figure 14 A. Tumor measurements were initiated on day 0. Tumor growth was monitored and the tumor volume was calculated for each mouse. Tumor growth curves for the respective treatments are shown in Figure 15Shown in B, where lines show the mean of mice undergoing treatment and error bars show SEM. KCL-HO-1 i in combination with gemcitabine can control tumor growth of this subcutaneous sarcoma tumor. **P < 0.01.

[0601] Anti-metastatic effect.

[0602] For tumor purposes, 4T1 mammary adenocarcinoma cells were implanted orthotopically in Balb / c mice (BioLasco, Taiwan, under license in Charles River Laboratories). A total of 2.5 x 10 5 cells were suspended in 100 μL of Roswell Park Memorial Institute medium (RPMI 1640 medium), then injected subcutaneously into the mammary fat pad of syngeneic female mice, then tumor growth was monitored and depicted in Figure 15 A. On day 12 after tumor injection, mice received KCL-HO-1 i (25 μMol / kg) or vehicle daily by intraperitoneal injection for 14 days. On day 26 after tumor injection, lungs were harvested and metastatic nodules were counted for KCL-HO-1 i and vehicle treated groups and represented in Figure 15 B. KCL-HO-1 i exhibited anti-metastatic effect in 4T1 mice compared to vehicle treated group (see References B), where dots represent individual mice, **P < 0.01. This model shows that primary tumor growth was not affected, so the lack of lung metastasis observed for KCL-HO-1 i vehicle treated group is not merely a reflection of lower tumor burden.

[0603] ​

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[0622] 20. K. M. 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)

[0623] All publications mentioned in the above specification are herein incorporated by reference. While the illustrative embodiments of the application have been described with reference to the accompanying drawings, it is to be understood that the application is not limited to the precise embodiments, and that various changes and modifications can be effected therein by one skilled in the art without departing from the scope of the application as defined by the appended claims and their equivalents.

Claims

1. A method for preparing a compound of formula (I) or its metal derivatives, salts, and solvates: The method includes the following steps: The following dicarboxylic acid compound: Or its salts and solvates are added to dialdehyde compounds of the following formula: Or its salt and solvate, and react to form a porphyrin diester compound of the following formula: Or its metal derivatives, salts and solvates; Each of R1, R2, R3, R4, R5, and R6 is independently selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", and C. 1-2 Haloalkyl, C 1-2 Alkylamines and C(O)CH3; and R7 and R8 are independently selected from C 1-6 Alkyl group; and R' and R" are each independently selected from H and C. 1-6 alkyl.

2. The method according to claim 1, wherein the method further comprises the following step: Hydrolysis of the porphyrin diester compound or its metal derivatives, salts and solvates provides a dicarboxylic acid of formula (I) or its metal derivatives, salts and solvates.

3. The method according to claim 1 or 2, wherein the preparation of the dicarboxylic acid compound further comprises the following step: Pyrrole A: or its salts and solvates With pyrrole B: Or its salts and solvates react; To form diester compounds of the following formula: or its salts and solvates, wherein R9 and R 10 It is independently selected from benzyl, p-methoxybenzyl and 2,4-dimethoxybenzyl.

4. The method according to claim 3, wherein the method further comprises the following step: Make the following diester compound: Its salts and solvates undergo deprotection reactions to form dicarboxylic acid compounds of the following formula: Or its salts and solvates.

5. The method according to any one of the preceding claims, wherein the method further comprises the following steps: The compound of formula (I) or its metal derivatives, salts and solvates are converted into metal derivatives, wherein the metal derivatives are tin compounds, and wherein the tin compounds are porphyrin dicarboxylic acids of formula (I) or their salts and solvates, tin methanesulfonate compounds, tin oxalate compounds, tin phosphate compounds or tin tartrate compounds.

6. The method according to any one of the preceding claims, wherein the method further comprises the following step: The compound of formula (I) or its metal derivative, salt, and solvate are converted into a metal derivative, wherein the metal derivative is a tin compound, and wherein the tin compound is: Or its salts and solvates.

7. The method according to any one of the preceding claims, wherein the method further comprises the following step: To convert a compound of formula (I) or its metal derivative salt and solvate into a tin (IV) phosphate salt of formula (II) or (III) or its salt and solvate:

8. A compound of formula (I) or a metal derivative salt thereof and a solvate thereof, wherein the compound is: Or its salts and solvates, wherein each of R1, R2, R3, R4, R5 and R6 is independently selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 Haloalkyl, C 1-2 Alkylamine and C(O)CH3; and R' and R” are each independently selected from H and C. 1-6 alkyl.

9. The compound according to claim 8, wherein the compound is a compound of formula (II) or (III): Or its salts and solvates, wherein each of R1, R2, R3, R4, R5 and R6 is independently selected from H, methyl, ethyl, halogen, methoxy, ethoxy, NR'R", C 1-2 Haloalkyl, C 1-2 Alkylamine and C(O)CH3; and R' and R” are each independently selected from H and C. 1-6 alkyl.

10. The compound of formula (II) or (III) according to claim 9, wherein R1, R3, R5, and R6 are methyl; and R2 and R4 are ethyl, said compound being capable of being represented by formula (VII) or (VIII) or its salts and solvates:

11. Use of the compound of formula (I) according to claim 8, 9 or 10, or its salts and solvates, in the treatment of proliferative and / or malignant diseases.

12. Use of the compound of formula (I) according to claim 11, or its salts and solvates, in the treatment of proliferative and / or malignant diseases, wherein the proliferative and / or malignant diseases are selected from breast cancer, lung cancer, brain cancer and central nervous system cancer, carcinoma, gastrointestinal cancer, hormone-induced cancer, leukemia, liver cancer, lymphoma, sarcoma and fibrosarcoma, skin cancer and melanoma, urinary tract cancer and reproductive cancer, and various other cancers.

13. Use of the compound of formula (I) according to claim 11 or 12, or its salts and solvates, in the treatment of proliferative and / or malignant diseases, wherein the compound is administered simultaneously or sequentially with one or more immunotherapeutic agents or other anticancer agents.

14. Use of the compound of formula (I) according to claim 8, 9 or 10, or its salts and solvates, in the treatment or inhibition of metastasis.

15. A pharmaceutical composition comprising a compound of formula (I) according to claim 8, 9 or 10, or a salt thereof and a solvate thereof, and a pharmaceutically acceptable excipient, carrier or diluent.

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