Swap70 inhibitors for use in the treatment of cancer

Abstract

The present disclosure relates to the use of compounds such as compounds of Formula (I) or a pharmaceutically acceptable salt, solvate and / or prodrug thereof in treating cancer and / or reduction and / or prevention of tumor invasion and / or metastasis.

Description

SWAP70 INHIBITORS FOR USE IN THE TREATMENT OF CANCERFIELD

[0001] The present disclosure relates to the use of compounds such as 3-(2-chlorophenyl)- 5-methyl-N-(6-nitrobenzo[d]thiazol-2-yl)isoxazole-4-carboxamide (B2), for example, in the treatment of cancer and / or reduction and / or prevention of tumor invasion and / or metastasis.BACKGROUND

[0002] By far the largest fraction of cancer-related death occurs through metastasis and thus, metastasis is the greatest challenge to cancer therapy. While new approaches have been established, such as immune checkpoint or cell-based therapies, additional new therapies are urgently required to solve this most pressing problem for human health.

[0003] Key steps in metastasis include the dissociation of tumor cells from the primary tumor, the entry into and migration through blood or lymph vessels, the exit from vessels, entry into tissues, and finally homing to a suitable tissue area to grow a metastatic tumor. This stepwise process, governed by multiple interactions, is reminiscent of Stephen Paget’s “seed and soil” theory of 1889 (Paget 1889). All these steps require dynamic rearrangements of the F-actin cytoskeleton of the tumor cell and thus proteins that govern F-actin dynamics. The highly active F-actin dynamics have been identified as a hallmark of aggressive and / or metastasising tumor cells, which form a range of F-actin-based structures such as lamellipodia, filopodia, and invadopodia to facilitate migration and invasion (Verschueren, Van der Taelen et al. 1994, Muller, Homey et al. 2001, Olson and Sahai 2009, Hanahan and Weinberg 2011, Numberg, Kitzing et al. 2011, Gross 2013, Steeg 2016, Lambert, Pattabiraman et al. 2017, Mondal, Di Martino et al. 2021).

[0004] In recent years, F-actin modulatory and accessory proteins increasingly attracted attention as targets in metastasis prevention and / or therapy (Numberg, Kollmannsperger et al. 2014, Huang, Hein et al. 2015, Yin, Ulloa et al. 2019, Barik, Sahay et al. 2022, Limaye, Whittaker et al. 2022). Other F-actin bundling proteins have been shown to be involved in tumor cell metastasis; notably Fascin has been reported to have an important role in promoting tumor cell invasion (Adams 2004, Machesky and Li 2010).

[0005] SWAP-70 was first described in 1998 as a protein involved in the isotype switch of immunoglobulins (Borggrefe, Wabl et al. 1998), and in 2000 as a protein which in murine B cells and in mice supports the production specifically of the IgE isotype of immunoglobulins (Borggrefe, Keshavarzi et al. 2001). In subsequent studies, it was shown that SWAP-70 has a key role in hematopoietic cell migration and adhesion, which are central to many inflammatory processes (Pearce, Angeli et al. 2006, Chopin, Quemeneur et al. 2010, Chopin, Chacon-Martinez et al. 2011, Ocana-Morgner, Reichardt et al. 2011, Pearce, Audzevich et al. 2011). The underlying mechanisms were then identified: in the cytoplasm and at cytoplasmic membranes SWAP-70 controls specific F-actin rearrangements in certain hematopoietic cells and thereby controls cell morphogenesis, mobility, adhesion and several specific functions that depend on the F-actin network), suggesting a role for SWAP-70 in tumor metastasis (Chacon-Martinez, Kiessling et al. 2013, Betaneli and Jessberger 2020).

[0006] In addition, SWAP-70 has been generally implicated in malignant diseases, in particular a role for SWAP-70 in several types of cancer in humans and other organisms has been disclosed (Heerema, Abbey et al. 2004, Fukui, Tanaka et al. 2007, Murugan, Ihara et al. 2008, Seol, Smith et al. 2009, Chiyomaru, Tatarano et al. 2011, Shu, Jing Yang et al. 2013, Chang, Shu et al. 2016, Kriplani, Duncan et al. 2019, Shi, Liu et al. 2019, Stanton, Gad et al. 2019, You, Zhang et al. 2019, Zhou, Shou et al. 2019).

[0007] Chemotherapy is a common approach for the treatment of cancer. However, the synthesis and / or identification of new, effective and safe chemotherapeutic agents remains a challenge. Reddy et al. disclose a series of pyrazolo-benzothiazole hybrids which were screened for their cytotoxic activity towards several cancer cell lines: colon (HT-29), prostate (PC-3), lung (A549), and glioblastoma (U87MC) (Reddy et al. 2019). Compounds with fluoro or chloro on the benzothiazole moiety displayed significant activity, with a compound with a chloro at that position being particularly potent towards all the tested cancer cell lines and considered by Reddy et al. to be a lead molecule for further development of VEGFR-2 inhibitors.

[0008] Canadian Patent Application No. 2,900,798 discloses a method for identifying a substance which inhibits the activity of SWAP-70, the method comprising: contacting at least one test substance with SWAP-70, detecting the degree of dimerization of SWAP-70, and selecting a test substance which inhibits the dimerization of SWAP-70. CA 2,900,798 alsogenerally discloses the use of a substance obtained by such a method for treatment of cancer.However, no specific substances have been disclosed in 2,900,798 for such treatment.

[0009] Accordingly, while SWAP-70 represents a potential target for treatment of cancer, there remains a need for compounds that inhibit the activity of SWAP-70 that would be useful as a potential therapy in such treatment.SUMMARY

[0010] An objective of the present invention is to reduce the mobility of cancer cells for example, to reduce tumor invasion and / or metastasis. B2 and other compounds of the present disclosure are predicted to act as inhibitors of SWAP-70 ’s interaction with F-actin.

[0011] Accordingly, the present disclosure includes a compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof for use in treating cancer:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; andn is an integer of from 0 to 4.

[0012] In an embodiment, the cancer is selected from prostate cancer, breast cancer, cervical cancer, thyroid cancer, colorectal cancer, stomach cancer, liver cancer, lung cancer, lymphoma, leukemia, glioblastoma, sarcoma and melanoma.

[0013] In an embodiment, the cancer comprises cancer stem cells.

[0014] The present disclosure also includes a compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof for use in reducing and / or preventing tumor invasion and / or metastasis:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is an integer of from 0 to 4.

[0015] In an embodiment, the tumor invasion and / or metastasis is in a subject having a primary cancer selected from prostate cancer, breast cancer, cervical cancer, thyroid cancer, colorectal cancer, stomach cancer, liver cancer, lung cancer, glioblastoma, sarcoma and melanoma. In another embodiment, the primary cancer comprises cancer stem cells.

[0016] In an embodiment, X2is O.

[0017] In an embodiment, m is 1.

[0018] In an embodiment, n is 1.

[0019] In an embodiment:R1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2is selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;R3is NO2;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is 1 ; and n is an integer of from 1 to 4.

[0020] In an embodiment:R4, R5, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; andR6is selected from H and Ci-ioalkyl.

[0021] In an embodiment, the compound of Formula I is a compound of Formula 1(a):1(a) wherein R1, R2, R3, X1, X2, X3, X4and X5are as described herein.

[0022] In an embodiment, the compound of Formula I is a compound of Formula I(a)(i):I(a)(i) wherein R1, R2, R3, X1, X3, X4and X5are as described herein.

[0023] In an embodiment, R3is NO2.

[0024] In an embodiment, R1is Ci-4alkyl. In another embodiment, R1is methyl.

[0025] In an embodiment, R2is halo. In another embodiment, R2is chloro.

[0026] In an embodiment, X1is N.

[0027] In am embodiment, X3is NH.

[0028] In am embodiment, X4is S.

[0029] In am embodiment, X5is N.

[0030] In an embodiment, the compound of Formula I has the structure:

[0031] In an embodiment, the compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof is the compound of Formula I.

[0032] The present disclosure also includes a pharmaceutical composition comprising a compound of the present disclosure for use in treating cancer.

[0033] In an embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0034] In an embodiment, the cancer is selected from prostate cancer, breast cancer, cervical cancer, thyroid cancer, colorectal cancer, stomach cancer, liver cancer, lung cancer, lymphoma, leukemia, glioblastoma, sarcoma and melanoma.

[0035] In an embodiment, the cancer comprises cancer stem cells.

[0036] Other features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating embodiments of the disclosure, are given by way of illustration only and the scope of the claims should not be limited by these embodiments but should rather be given the broadest interpretation consistent with the description as a whole.BRIEF DESCRIPTION OF THE DRAWINGS

[0037] The embodiments of the disclosure will now be described in greater detail with reference to the attached drawings, in which:

[0038] FIG. 1 shows inhibition of SWAP-70 dimerization in living cells by 3-(2- chlorophenyl)-5-methyl-N-(6-nitrobenzo[d]thiazol-2-yl)isoxazole-4-carboxamide (B2) in comparison to control. The left plot shows inhibition of the fluorescence-activated cell sorting- fluorescence resonance energy transfer (FACS -FRET) signal (FRET efficiency) and thus ofSWAP-70 dimerization by three different concentrations of B2, measured at 20 minutes after stimulation of the cells; and the right plot shows inhibition by 50 μM B2 at earlier time intervals after stimulation. B2 was added to the cells 15 minutes prior to stimulation.

[0039] FIGs. 2-4 show the binding of B2 to the target molecule (SWAP-70) in solution assessed in the microscale thermophoresis (MST) assay. FIG. 2 shows the mobility of SWAP- 70 at different time points up to 20 minutes after incubation of SWAP-70 with B2; note the change in signal of the complex compared to the target (SWAP-70). FIG. 3 shows control measurements of the fluorescence signals at the 14-15 minute interval after incubation of SWAP-70 with B2. FIG. 4 shows quantification of the mobility shift and thus binding within the same interval, demonstrating significant binding (n = 4).

[0040] FIG. 5 shows the binding of B2 to SWAP-70 (lower curve) in comparison to SWAP-70 only (upper curve) in the nanoDSF (differential scanning fluorimetry) assay.

[0041] FIG. 6 shows inhibition of 4T1 breast carcinoma tumor cell growth by B2 in a soft agar colony formation assay in comparison to control. The inhibition of 4T1 breast carcinoma tumor cell growth was measured two weeks after seeding of cells and addition of, from left to right: 0 μM (dimethylsulfoxide, DMSO), 50 μM, 20 μM and 10 μM B2. The entire cell culture plates (upper row) and a representative magnified area (lower row) are shown. The colony size is also strongly reduced in presence of B2 as is obvious from the magnified panel.

[0042] FIG. 7 is a plot showing the quantified results of three experiments of inhibition of 4T1 breast carcinoma tumor cell growth by 50 μM, 20 μM and 10 μM B2 in a soft agar colony formation assay in comparison to control as number of colonies.

[0043] FIG. 8 shows inhibition of 4T1 mouse breast carcinoma cell migration by B2 through a membrane in a Transwell set-up. The average number of 4T1 breast carcinoma tumor cells was determined 24 hours after insertion of cells and addition of from left to right in graph: 0 μM (DMSO), 20 μM or 50 μM B2; average of three experiments.

[0044] FIG. 9 shows images showing representative fields of view of migrated cells for 0 μM (left image) and 50 μM B2 (right image) in the Transwell set-up described for FIG. 8.

[0045] FIG. 10 shows inhibition of 4T1 breast carcinoma cells migration by B2 in a wound healing assay. The percentage of “wound” area covered by 4T1 breast carcinoma tumor cellswas determined 18 hours after addition of three different concentrations of B2, from left to right: 0 μM (DMSO), 50 μM, 20 μM or 10 μM B2. Average of at least 5 experiments.

[0046] FIG. 11 shows exemplary images showing a “wound” within a cell layer in the wound healing assay described for FIG. 10, for control (DMSO, left image) and 50 μM B2 (right image). The white arrow points to the edge of the wound where cells are actively migrating into the free space only in the control (DMSO, left image).

[0047] FIG. 12 shows inhibition of 3D spheroid formation of 4T1 breast carcinoma cells by B2 for, from left to right: no B2 (DMSO), and 50 μM, 10 μM, 1 μM or 0.1 μM B2. The average spheroid area (left graph) and the average fluorescence of the stained cells (right graph) are shown from 3 independent experiments.

[0048] FIG. 13 shows images presenting exemplary spheroids from the examples relating to inhibition of 3D spheroid formation described for FIG. 12.

[0049] Fig. 14 shows inhibition of tumor growth of 4T1 breast carcinoma cells in mice by B2 (B2-treated) compared to a control (no B2, DMSO), left: tumor volume, right: tumor weight, as average from 10 mice each group.DETAILED DESCRIPTIONI. Definitions

[0050] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the disclosure herein described for which they would be understood to be suitable by a person skilled in the art.

[0051] As used herein, the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “include” and “includes”) or “containing” (and any form of containing, such as “contain” and “contains”), are inclusive or open-ended and do not exclude additional, unrecited elements or process / method steps. As used herein, the word “consisting” and its derivatives, are intended to be close ended terms that specify the presence of stated features, elements, components, groups, integers, and / or steps, and also exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The term “consisting essentially of’, as used herein, is intended to specify the presenceof the stated features, elements, components, groups, integers, and / or steps as well as those that do not materially affect the basic and novel characteristic(s) of these features, elements, components, groups, integers, and / or steps.

[0052] As used herein, terms of degree such as “substantially”, “about” and “approximately” mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be construed as including a deviation of at least ±5% or at least ±10% of the modified term if this deviation would not negate the meaning of the term it modifies.

[0053] As used in this disclosure, the singular forms “a”, “an” and “the” include plural references unless the content clearly dictates otherwise.

[0054] The term “and / or” as used herein means that the listed items are present, or used, individually or in combination. In effect, this term means that “at least one of’ or “one or more” of the listed items is present or used.

[0055] The term “suitable” as used herein means that the selection of the particular compound and / or conditions would depend on the specific synthetic manipulation to be performed, and / or the identity of the compound(s) to be transformed, but the selection would be well within the skill of a person skilled in the art. All synthetic method steps described herein are to be conducted under conditions sufficient to provide the product shown. A person skilled in the art would understand that all reaction conditions, including, for example, reaction solvent or lack thereof, reaction time, reaction temperature, reaction pressure, reactant ratio and whether or not the reaction should be performed under an anhydrous or inert atmosphere, can be varied to optimize the yield of the desired product and it is within their skill to do so.

[0056] The expression “proceed to a sufficient extent” as used herein with reference to the reactions or method steps disclosed herein means that the reactions or method steps proceed to am extent that conversion of the starting material or substrate to product is maximized. Conversion may be maximized when greater than about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100% of the starting material or substrate is converted to product.

[0057] The term “subject” as used herein includes all members of the animal kingdom including mammals, and optionally refers to humans. In an embodiment, the subject is human.

[0058] The term “halo” as used herein refers to a halogen atom and includes F, Cl, Br and I. In an embodiment of the present disclosure, halo is chloro.

[0059] The term “alkyl” as used herein, whether it is used alone or as part of another group, means straight or branched chain, saturated alkyl groups. The number of carbon atoms that are possible in the referenced alkyl group are indicated by the numerical prefix “Cn1-n2”. For example, the term Ci-ioalkyl means an alkyl group having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.

[0060] The term “aryl” as used herein, whether used alone or as part of another group, refers to groups that contain at least one aromatic ring. When an aryl group contains more than one aromatic ring the term “aryl” as used herein includes condensed aromatic systems. In an embodiment, the aryl group contains from 6, 9, 10 or 14 atoms, such as phenyl, naphthyl, indanyl or anthracenyl. The number of carbon atoms that are possible in the referenced aryl group are indicated by the numerical prefix “Cn1-n2”. For example, the term Ce-ioaryl means an aryl group having 6, 7, 8, 9 or 10 carbon atoms.

[0061] The term “heteroaryl” as used herein, whether used alone or as part of another group, refers to an aromatic, ring-containing group having one or more multivalent heteroatoms (for example, heteroatoms independently selected from N, O and S), as a part of the ring structure. In an embodiment of the present disclosure, the heteroaryl includes at least 5 and up to 20 atoms in the ring(s). Heteroaryl groups may contain more than one ring.

[0062] The term “substituted” as used herein refers to a structure, molecule or group in which one or more available hydrogen atoms are replaced with one or more other chemical groups. A person skilled in the art would readily appreciate that the number of possible substituents would depend, for example, on the particular structure, molecule or group and / or the number of available hydrogen atoms therein. For example, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 5, 4, 3, 2 and / or 1 available hydrogen atoms may be replaced with the one or more other chemical groups. The term “available hydrogen atoms” as used herein refers to hydrogens that would be known to a person skilled in the art to be capable of replacement by a suitable substituent. In an embodiment, the substituents are independently selected from one or more of C1-4alkyl, aryl, heteroaryl, halo, OR9, NO2 and SO2R9, wherein R9is selected from H, C1-4alkyl, aryl and heteroaryl.

[0063] The term “B2” as used herein refers to the compound 3-(2-chlorophenyl)-5-methyl-N-(6-nitrobenzo[d]thiazol-2-yl)isoxazole-4-carboxamide, having the chemical structure:

[0064] The terms “compounds of the disclosure”, “compounds of the present disclosure” and the like as used herein include compounds of Formula I and pharmaceutically acceptable salts, solvates and / or prodrugs thereof, including compounds of Formula 1(a) and pharmaceutically acceptable salts, solvates and / or prodrugs thereof, and including compounds of Formula I(a)(i) and pharmaceutically acceptable salts, solvates and / or prodrugs thereof as well as B2 and pharmaceutically acceptable salts, solvates and / or prodrugs thereof.

[0065] The term “pharmaceutically acceptable” means compatible with the treatment of subjects, for example, mammals such as humans.

[0066] The term “pharmaceutically acceptable salt” as used herein means an acid addition salt or a base addition salt that is compatible with the treatment of subjects.

[0067] An “acid addition salt that is compatible with the treatment of subjects” is any non- toxic inorganic or organic salt of any basic compound. Basic compounds that form an acid addition salt include, for example, compounds comprising an amine group susceptible to protonation. Illustrative inorganic acids which form suitable salts include hydrochloric, hydrobromic, sulfuric and phosphoric acids, as well as metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Illustrative organic acids that may form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartan c, citric, ascorbic, maleic, benzoic, phenylacetic, cinnamic and salicylic acids, as well as sulfonic acids such as p-toluene sulfonic and methanesulfonic acids. The selection of a suitable salt can be made by a person skilled in the art. The formation of a desired acid addition salt is, for example, achieved using standard techniques. For example, in an embodiment of the present disclosure, the neutral compoundis treated with the desired acid in a suitable solvent and the salt which is thereby formed then isolated by filtration, extraction and / or any other suitable method.

[0068] A “base addition salt that is compatible with the treatment of subjects” is any non- toxic inorganic or organic salt of any acidic compound. Acidic compounds that form a base addition salt include, for example, compounds comprising a sulfonic acid group. Inorganic bases that may form suitable salts include, without limitation, lithium, sodium, potassium, calcium, magnesium or barium hydroxide. Organic bases that may form suitable salts include, without limitation, aliphatic, alicyclic or aromatic organic amines such as methylamine, trimethylamine and picoline or ammonia. The selection of a suitable salt can be made by a person skilled in the art. The formation of a desired base addition salt is, for example, achieved using standard techniques. For example, in an embodiment of the present disclosure, the neutral compound is treated with the desired base in a suitable solvent and the salt which is thereby formed then isolated by filtration, extraction and / or any other suitable method.

[0069] The term “solvate” as used herein in reference to a compound of the disclosure refers to a complex formed between the compound and a solvent from which the compound is precipitated or in which the compound is made. Accordingly, the term “solvate” as used herein means a compound of the disclosure, wherein molecules of a suitable solvent are incorporated in the crystal lattice. Examples of suitable solvents are ethanol, water and the like. When water is the solvent, the complex is optionally referred to as a “hydrate”. The formation of solvates will vary depending on the compound and the solvate. In general, solvates are formed by dissolving the compound in an appropriate solvent and isolating the solvate by cooling or using an antisolvent and / or reducing and / or evaporating the solvent. The solvate is typically dried or azeotroped under ambient conditions. The selection of suitable conditions to form a particular solvate can be made by a person skilled in the art.

[0070] The term “prodrug” as used herein in reference to a compound of the disclosure refers to a derivative of the compound that reacts under biological conditions to provide the compound.

[0071] The term “cancer” as used herein refers to a disease involving abnormal cell growth with the potential to invade and / or spread into other parts of the body such as healthy tissue and that is treatable via inhibition of SWAP-70. The term “cancer” as used herein includes malignant neoplasms (malignant or cancerous tumors) and hematologic cancers thatare treatable via inhibition of SWAP-70 and may include, for example prostate cancer, breast cancer, cervical cancer, thyroid cancer, colorectal cancer, stomach cancer, liver cancer, lung cancer, lymphomas, leukemias, glioblastomas, cancer stem cells, sarcomas and melanoma. Many cancers spread beyond their initial boundaries and invade adjoining parts of the body and / or spread to other tissues. Metastasis is a major cause of death from cancer. Metastasis or metastatic cancer occurs when cancer cells break off from the original (primary) tumor and spread to other areas of the body via a process comprising migration through blood or lymph vessels.

[0072] The terms “inhibiting”, “inhibited” and the like as used herein, e.g., in respect to inhibiting SWAP-70 means an inhibition that causes a therapeutic effect in respect to the treatment of cancer and may be any detectable inhibition in the presence of a compound of the present disclosure in comparison to the same conditions without the compound.

[0073] The terms “reducing” and the like as used herein, e.g., in respect to tumor invasion and / or metastasis may be any detectable reduction in the presence of a compound of the present disclosure in comparison to the same conditions without the compound.

[0074] The terms “preventing” and the like as used herein, e.g., in respect to tumor invasion and / or metastasis means a prophylactic treatment to prevent tumor invasion and / or metastasis.

[0075] The terms “to treat”, “treating” and “treatment” and the like as used herein and as is well understood in the art, means an approach for obtaining beneficial or desired results, including clinical results. For example, in the context of treating cancer, beneficial or desired clinical results include, but are not limited to alleviation or amelioration of one or more symptoms of the cancer, diminishment of the extent of the cancer, stabilized (i.e., not worsening) of the cancer, delay or slowing of the progression of the cancer, amelioration or palliation of the disease state of the cancer, diminishment of the reoccurrence of the cancer, and / or remission (whether partial or total) of the cancer, whether detectable or undetectable. “To treat”, “treating” and “treatment” and the like as used herein also include prophylactic treatment. For example, a subject with early stage cancer is treated to prevent progression or alternatively a subject in remission is treated to prevent recurrence.

[0076] The compounds of the disclosure are, for example, administered to the subject or used in an “effective amount”. As used herein, the term “effective amount” and the like means an amount effective, at dosages and for periods of time necessary to achieve a desired result. For example, in the context of treating cancer, an effective amount of a compound administered or used is an amount that, for example, reduces the size of a tumor of the cancer and / or amount of metastases of a cancer compared to the cancer without administration or use of the compound. Effective amounts may vary according to factors such as the disease state, age, sex, weight and / or species of the subject. The amount of a given compound that will correspond to such an amount will vary depending upon various factors, such as the given compound, the pharmaceutical formulation, the route of administration or use, the type of cancer being treated, the identity of the subject being treated, and the like, but can nevertheless be routinely determined by one skilled in the art.II. Methods of Treatment and Uses

[0077] The compound B2 was selected in vitro for inhibition of SWAP-70 and subsequently shown to block SWAP-70 dimerization in living cells. B2 has been shown to directly bind to SWAP-70 protein. B2 has further been shown to block SWAP-70 dimerization inside cells, which is essential for its F-actin modulatory activity. In addition, B2 has been shown in cell culture assays to inhibit several tumor invasion and metastasis-related activities. Accordingly, B2 and / or other compounds of the present disclosure may be useful to treat cancer treatable by inhibiting SWAP-70.

[0078] Accordingly, the present disclosure includes a method of treating cancer, the method comprising administering an effective amount of a compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof to a subject in need thereof:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is an integer of from 0 to 4.

[0079] The present disclosure also includes the use of a compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof for treating cancer:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is an integer of from 0 to 4.

[0080] The present disclosure also includes a use of a compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof for preparation of a medicament for treating cancer:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is an integer of from 0 to 4.

[0081] The present disclosure also includes a compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof for use in treating cancer:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is an integer of from 0 to 4.

[0082] In an embodiment, the cancer is a primary cancer and the treatment reduces and / or prevents metastasis from the primary cancer.

[0083] In another embodiment, the cancer comprises a tumor and the treatment reduces and / or prevents tumor invasion.

[0084] In am embodiment, the cancer is selected from prostate cancer, breast cancer, cervical cancer, thyroid cancer, colorectal cancer, stomach cancer, liver cancer, lung cancer, lymphoma, leukemia, glioblastoma, sarcoma and melanoma.

[0085] In another embodiment, the cancer comprises cancer stem cells.

[0086] The present disclosure also includes a method of reducing and / or preventing tumor invasion and / or metastasis, the method comprising administering an effective amount of a compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof to a subject in need thereof:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is am integer of from 0 to 4.

[0087] The present disclosure also includes a use of a compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof for reducing and / or preventing tumor invasion and / or metastasis:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is an integer of from 0 to 4.

[0088] The present disclosure also includes a use of a compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof for preparation of a medicament for reducing and / or preventing tumor invasion and / or metastasis:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is an integer of from 0 to 4.

[0089] The present disclosure also includes a compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof for use in reducing and / or preventing tumor invasion and / or metastasis:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, Ci-walkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is an integer of from 0 to 4.

[0090] In an embodiment, the tumor invasion and / or metastasis is in a subject having a primary cancer selected from prostate cancer, breast cancer, cervical cancer, thyroid cancer, colorectal cancer, stomach cancer, liver cancer, lung cancer, glioblastoma, sarcoma and melanoma.

[0091] In another embodiment, the primary cancer comprises cancer stem cells.

[0092] In an embodiment, m is an integer of from 1 to 5, 1 to 4, 1 to 3 or 1 to 2. In another embodiment, m is 4, 3, 2 or 1. In an embodiment, m is not 0. In an embodiment, m is not 2. In an embodiment, m is 1. In an embodiment, when m is 0, R3is selected from substituted or unsubstituted aryl, heteroaryl, halo, NO2and SO2R5. In another embodiment, m is 1 and R2is halo, n is an integer of from 1 to 4, and R3is selected from substituted or unsubstituted aryl, heteroaryl and NO2. In a further embodiment, when m is 2, R2is selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, OR4, NO2and SO2R5.

[0093] In an embodiment, n is an integer of from 1 to 4, 1 to 3 or 1 to 2. In another embodiment, n is 3, 2 or 1. In a further embodiment, n is 1.

[0094] In an embodiment:R1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2is selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2and SO2R5;R3is NO2;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is 1 ; and n is an integer of from 1 to 4.

[0095] In another embodiment:R4, R5, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; andR6is selected from H and C1-10alkyl.

[0096] In an embodiment, the compound of Formula I is a compound of Formula 1(a):whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH; andR4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; orR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2is selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2and SO2R5;R3is N02;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH; andR4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl.

[0097] In an embodiment, the compound of Formula I is a compound of Formula 1(a) (i):I(a)(i) 5 whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH; andR4, R5, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; orR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2is selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2and SO2R5;R3is NO2;X1is selected fromX3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2and NR8;X5is selected from N and CH; andR4, R5, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl.

[0098] In an embodiment, R1is H. In another embodiment, R1is selected from C1-10alkyl, substituted or unsubstituted aryl and heteroaryl. In a further embodiment, R1is C1-10alkyl. In another embodiment, R1is Ci-salkyl. In a further embodiment, R1is Ci-4alkyl. In another embodiment, R1is methyl.

[0099] In an embodiment, R2is halo. In another embodiment, R2is chloro.

[0100] In an embodiment, R3is NO2. In an embodiment, R3is not SO2R5.

[0101] In an embodiment, R4is H. In another embodiment, R4is selected from C1-10alkyl, substituted or unsubstituted aryl and heteroaryl. In a further embodiment, R4is C1-10alkyl. In another embodiment, R4is substituted or unsubstituted aryl. In an embodiment, R4is heteroaryl.

[0102] In an embodiment, R5is H. In another embodiment, R5is selected from C1-10alkyl, substituted or unsubstituted aryl and heteroaryl. In a further embodiment, R5is C1-10alkyl. In another embodiment, R5is substituted or unsubstituted aryl. In an embodiment, R5is heteroaryl.

[0103] In an embodiment, R6is H. In another embodiment, R6is selected from C1-10alkyl, substituted or unsubstituted aryl and heteroaryl. In a further embodiment, R6is C1-10alkyl. In another embodiment, R6is substituted or unsubstituted aryl. In an embodiment, R6is heteroaryl.

[0104] In an embodiment, R7is H. In another embodiment, R7is selected from C1-10alkyl, substituted or unsubstituted aryl and heteroaryl. In a further embodiment, R7is C1-10alkyl. In another embodiment, R7is substituted or unsubstituted aryl. In an embodiment, R7is heteroaryl.

[0105] In am embodiment, R8is H. In another embodiment, R8is selected from C1-10alkyl, substituted or unsubstituted aryl and heteroaryl. In a further embodiment, R8is C1-10alkyl. In another embodiment, R8is substituted or unsubstituted aryl. In an embodiment, R8is heteroaryl.

[0106] In an embodiment, X1is N.

[0107] In an embodiment, X2is O.

[0108] In an embodiment, X3is NR7. In another embodiment, X3is NH.

[0109] In an embodiment, X4is S.

[0110] In an embodiment, X5is N.

[0111] In an embodiment, X1is N, X2is O, X3is NH, X4is S and / or X5is N.

[0112] In an embodiment, the compound of Formula I has the structure:

[0113] In an embodiment, of the compounds of Formula I: when m is 0, R3is selected from substituted or unsubstituted aryl, heteroaryl, halo, NO2 and SO2R5; when m is 1 and R2is halo, n is an integer of from 1 to 4, and R3is selected from substituted or unsubstituted aryl, heteroaryl and NO2; and when m is 2, R2is selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, OR4, NO2 and SO2R5.

[0114] In an embodiment, the compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof is the compound of Formula I; i.e., is not the pharmaceutically acceptable salt, solvate and / or prodrug of the compound of Formula I.

[0115] In an embodiment, the subject is a human.

[0116] In am embodiment, the compounds of the disclosure are for administration or use in the form of a pharmaceutical composition comprising one or more compounds of the present disclosure and optionally a pharmaceutically acceptable carrier. In another embodiment, the pharmaceutical composition comprises the pharmaceutically acceptable carrier. In another embodiment of the present disclosure, the pharmaceutical composition comprises a pharmaceutically acceptable thinner or carrier, and at least one further component selected from the group consisting of diluents, fillers, binders, and other excipients or stabilizers.

[0117] The present disclosure also includes a pharmaceutical composition as described herein for use in any of the methods of treating or uses as described herein. For example, the present disclosure includes a method of treating cancer as described herein, comprising administering an effective amount of a pharmaceutical composition as described herein to a subject in need thereof; and a method of reducing and / or preventing tumor invasion and / or metastasis as described herein, the method comprising administering an effective amount of a pharmaceutical composition as described herein to a subject in need thereof; as well as the corresponding uses. It will be appreciated by a person skilled in the art that the embodiments of such methods and uses can be varied as described herein in respect to the methods and uses in respect to the compounds of the present disclosure described hereinabove.

[0118] The compounds of the disclosure can be administered to a subject or used in a variety of forms depending on the selected route of administration or use, as will be understood by those skilled in the art. In an embodiment, the one or more compounds of the disclosure are administered to the subject, or used, by oral (peroral) or parenteral (including injection (e.g., intravenous (i.v.), intramuscular (i.m.), intraperitoneal (i.p.), subcutaneous (s.c.) and intrathecal (i.t.) forms of administration or use), infusion, via implant, inhalational, intranasal, sublingual, buccal, rectal, vaginal, topical, transdermal, ocular and otic administration or use and the compound(s) formulated accordingly. For example, the compounds of the disclosure are administered or used in an injection, in a spray, in a tablet / caplet, in a powder, topically, in a gel, in drops, by a patch, by an implant, by a slow-release pump or by any other suitable method of administration or use, the selection of which can be made by a person skilled in the art.

[0119] In an embodiment of the present disclosure, the one or more compounds of the present disclosure are administered or used parenterally. Pharmaceutical forms suitable for injectable administration or use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. A person skilled in the art would know how to prepare suitable formulations. In an embodiment, the one or more compounds of the present disclosure are administered or used via injection. In another embodiment, the one or more compounds of the present disclosure are orally administered or used, for example, with an inert diluent or with an assimilable edible carrier, or enclosed in hard or soft shell gelatin capsules, or compressed into tablets, or incorporateddirectly with the food of the diet. In an embodiment, for oral therapeutic administration or use, the one or more compounds of the disclosure are incorporated with excipient and administered or used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Oral dosage forms also include modified release, for example immediate release and timed-release, formulations. In an embodiment, the one or more compounds of the present disclosure are administered via inhalation.

[0120] Treatment methods or uses comprise administering to a subject or use of an effective amount of one or more compounds of the disclosure, optionally consisting of a single administration or use, or alternatively comprising a series of administrations or uses. For example, the compounds of the disclosure are administered or used at least once a week. However, in another embodiment, the compounds are administered to the subject or used from one time per three weeks, or one time per week to once daily for a given treatment or use. In another embodiment, the compounds are administered or used 2, 3, 4, 5 or 6 times daily. The length of the treatment period, the intervals, or use depend on a variety of factors, such as the kind of cancer, the stage of cancer, the age of the subject, the concentration of the one or more compounds in a formulation, the activity of the compounds of the present disclosure, and / or a combination thereof. It will also be appreciated that the effective amount of a compound used for the treatment or use may increase or decrease over the course of a particular treatment regime or use. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In some instances, chronic administration or use is required. For example, the one or more compounds of the present disclosure are administered or used in an amount and for duration sufficient to treat the subject.

[0121] The extent and / or undesirable clinical manifestations of the cancer are optionally lessened (palliated) and / or the time course of the progression is slowed or lengthened, as compared to not treating the cancer.

[0122] The one or more compounds of the disclosure may be administered or used alone or in combination with other cancer therapies, including in combination with additional therapeutic agents useful for treating cancer. In an embodiment, the one or more compounds of the disclosure are administered or used in combination with at least one further cancer therapy. In an embodiment, the at least one further cancer therapy is selected from surgical removal,radiation therapy, photodynamic therapy, external hyperthermia, stem cell transplant, bone marrow transplant, an additional therapeutic agent useful for treating cancer and combinations thereof. In an embodiment, the additional therapeutic agent useful for treating cancer is selected from a chemotherapeutic agent, an angiogenesis inhibitor, a therapeutic agent useful for hormone therapy, an immunotherapeutic agent, a therapeutic agent useful for targeted therapy and combinations thereof. In an embodiment, the at least one further cancer therapy is an immunotherapy. It will be appreciated by a person skilled in the art that the selection of a suitable cancer therapy will depend, for example, on the nature of the cancer. Such administration or use of the one or more compounds of the disclosure in combination with other cancer therapies may include simultaneous and sequential treatment. Thus, the compounds of the disclosure may optionally be administered or used before, after or simultaneously with the at least one other cancer therapy. For example, when administered or used in combination with other known therapeutic agents, it is an embodiment that the one or more compounds of the disclosure are administered or used contemporaneously with those therapeutic agents. As used herein the term “contemporaneous” in reference to administration of two substances to a subject or use means providing each of the two substances so that they are both biologically active in the individual at the same time. The exact details of the administration or use will depend on the pharmacokinetics of the two substances in the presence of each other and can include administering or using the two substances within a few hours of each other, or even administering or using one substance within 24 hours of administration or use of the other if the pharmacokinetics are suitable. Design of suitable dosing regimens is routine for one skilled in the art. In particular embodiments, two substances will be administered or used substantially simultaneously, i.e., within minutes of each other, or in a single composition that contains both substances. It is a further embodiment that a combination of the two substances is administered to a subject or used in a non-contemporaneous fashion.

[0123] The dosage of compounds of the disclosure can vary depending on many factors such as the pharmacodynamic properties of the compound, the mode of administration or use, the age, health and weight of the subject, the kind of cancer, the stage of cancer, the frequency of the treatment or use and the type of concurrent treatment or use, if any, and the clearance rate of the compound in the subject. One of skill in the art can determine the appropriate dosage based on the above factors. In an embodiment, the compounds of the disclosure are administeredor used initially in a suitable dosage that is optionally adjusted as required, depending on the clinical response. As a representative example, oral dosages of one or more compounds of the disclosure will range from less than 1 mg per day to 1000 mg per day for a human adult or an animal. In an embodiment, the compounds of the disclosure are administered or used in a single daily dose or the total daily dose may be divided into two, three or four daily doses.III. Preparation of Compounds of the Present Disclosure

[0124] The compounds of the present disclosure can be prepared by various methods, the selection of which can be readily made by a person skilled in the art. Suitable techniques for work-up and / or purification of intermediates and final products can also be selected by the skilled person. Compounds used in the synthesis of the compounds may be available from commercial sources such as but not limited to Sigma-Aldrich, Acros Organics, Lach-Ner or TCI or alternatively may be prepared by a suitable method know to those skilled in the art.

[0125] For example, in an embodiment wherein in the compounds of the present disclosure, X1is N, X2is O and X3is NH, the compounds are generally prepared according to a method illustrated in Scheme 1. Variables in Scheme 1 and the following embodiments of the exemplary method for preparation are as defined herein for the compounds of the present disclosure unless otherwise specified and A+is a suitable alkali metal ion such as sodium ion (Na+). A person skilled in the art could readily vary the method to prepare, for example, compounds having X1, X2and / or X3as otherwise defined herein for the compounds of the present disclosure.

[0126] In an embodiment, as shown in Scheme 1, the method comprises reaction of an optionally substituted benzaldehyde of Formula II with a suitable reagent to prepare the oxime of Formula III followed by chlorination of the oxime of Formula III to prepare the chloro-oxime of Formula IV, cyclization of the chloro-oxime of Formula IV and a suitable alkali metal salt (e.g., a sodium salt) of an acetoacetate of Formula V followed by hydrolysis of the methyl ester to prepare the carboxylic acid of Formula VI, which is amide-coupled with a compound of Formula VII (e.g., a 2-amino-benzothiazole derivative) to prepare the compound of Formula I.

[0127] In an embodiment, the reaction of the optionally substituted benzaldehyde of Formula II with a suitable reagent to prepare the oxime of Formula III comprises reacting the optionally substituted benzaldehyde of Formula II with hydroxylamine hydrochloride in the presence of a suitable base such as sodium hydroxide in a suitable solvent or mixture thereof such as a mixture of water and ethanol for a time and at a temperature for the conversion of the optionally substituted benzaldehyde of Formula II to the oxime of Formula III to proceed to a sufficient extent.

[0128] In an embodiment, the chlorination of the oxime of Formula III to the chloro-oxime of Formula IV comprises reaction of the oxime of Formula III with a suitable chlorinating reagent (e.g., N-chlorosuccinimide) in a suitable solvent or mixture thereof such as N,N- dimethylformamide (DMF) for a time and at a temperature for the conversion of the oxime of Formula III to the chloro -oxime of Formula IV to proceed to a sufficient extent. In an embodiment, the N-chlorosuccinimide is added to a solution of the oxime of Formula III in the suitable solvent or mixture thereof such as DMF portion-wise, for example, in 10, 9, 8, 7, 6, 5, 4, 3 or 2 portions. In another embodiment, the reaction comprises induction with HCl(g).

[0129] In an embodiment, the cyclization of the chloro-oxime of Formula IV and the suitable alkali metal salt of an acetoacetate of Formula V comprises reaction of the chloro- oxime of Formula IV and the suitable alkali metal salt of the acetoacetate of Formula V in the presence of a suitable base (e.g., NaOH) in a suitable solvent or mixture thereof such as methanol for a time and at a temperature for the conversion of the chloro-oxime of Formula IV and the suitable alkali metal salt of an acetoacetate of Formula V to the methyl ester of the carboxylic acid of Formula VI to proceed to a sufficient extent. In an embodiment, the alkali metal salt of the acetoacetate of Formula V is a sodium salt. In an embodiment, the suitable alkali metal salt of the acetoacetate of Formula V is added to a solution of the chloro-oximeof Formula IV in the suitable solvent or mixture thereof such as methanol portion -wise, for example, in 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3 or 2 portions. In an embodiment, the hydrolysis comprises reaction of the methyl ester of the carboxylic acid of Formula VI with a suitable hydrolyzing agent e.g., a solution of a suitable base such as NaOH in water for a time and at a temperature for the conversion of the methyl ester of the carboxylic acid of Formula VI to the carboxylic acid of Formula VI to proceed to a sufficient extent. In an embodiment, the hydrolysis is carried out in situ; e.g., without work-up, isolation and / or purification of the methyl ester of the carboxylic acid of Formula VI prior to the hydrolysis.

[0130] In an embodiment, the amide coupling of the carboxylic acid of Formula VI with the compound of Formula VII (e.g., the 2-amino-benzothiazole derivative) comprises addition of triethylamine and diisopropylcarbodiimide (DIC) to a mixture of the carboxylic acid of Formula VI, the compound of Formula VII and ethyl cyanohydroxyiminoacetate (Oxyma), in a suitable solvent or mixture thereof such as dimethylformamide and reacting for a time and at a temperature for the reaction of the carboxylic acid of Formula VI with the compound of Formula VII to proceed to a sufficient extent, followed by a suitable work-up.

[0131] The following are non-limiting examples of the present disclosure:EXAMPLESExample 1: Preparation of 3-(2-chlorophenyl)-5-methyl-N-(6-nitrobenzo[d]thiazol-2- yl)isoxazole-4-carboxamide (“B2”)I. Synthesis a) Synthesis of 2-chlorobenzaldehyde oxime

[0132] To a stirred mixture of 2-chlorobenzaldehyde (100 g, 0.711 mol) in water (100 mL) and 96% ethanol (250 mL) cooled to 5 °C by ice bath, hydroxylamine hydrochloride (54.5 g, 0.784 mol) was added. The solution was stirred at 5 °C and 50% NaOH (95 mL) was added slowly, turning the mixture strongly alkaline as indicated by litmus paper. The mixture was allowed to warm up to room temperature (r.t.), stirred for 1 h, acidified with concentrated HC1(25mL) to strongly acidic as indicated by litmus paper, and extracted with CH2CI2 (3x250 mL). The combined organic extracts were washed with water (2x250 mL), brine (2x250 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give a crystallizing oil. Yield: 109.0 g (98%). liquid chromatography-mass spectrometry (LC-MS): MS (electrospray ionization; ESI) calcd. for: C7H7CINO [M+H]+156.01; found: 155.99, retention time (tn) = 2.67 min. b) Synthesis of 2-chloro-N-hydroxybenzimidoyl chloride

[0133] To a stirred solution of the 2-chlorobenzaldehyde oxime (100 g, 0.64 mol) inN,N- dimethylformamide (DMF; 630 mL) was added one-fifth portion (17.1 g, 0.13 mol) of N- chlorosuccinimide (NCS). To induce the reaction, HCl(g) from an HC1 generator was bubbled through the reaction mixture with vigorous stirring until the temperature started to increase. The temperature of the solution rose to 35 °C and was kept below this value by periodical ice bath cooling. The rest of the NCS (68.4 g, 0.51 mol) was added in 4 portions. The time interval between portions was 15 min and then the reaction mixture was stirred for 0.5 h. The solution was concentrated in vacuo, and the residue was dissolved in methyl / -butyl ether (MTBE; 600 mL), washed with water (2x500 mL), brine (1x500 mL), dried over anhydrous sodium sulfate (25 g), filtered, and concentrated in vacuo to give a yellowish oil. Yield: 116.8 g (96%). LC- MS: MS (ESI) calcd. for: C7H6CI2NO [M+H]+189.97; found: 190.00, tR= 3.14 min. c) Synthesis of 3-(2-chlorophenyl)-5-methylisoxazole-4-carboxylic acid

[0134] To a solution of 2-chloro-N-hydroxybenzimidoyl chloride (121.76 g, 0.64 mol) in methanol (900 mL), cooled between -15 to -20 °C, was added NaOH (1.3 g, 0.032 mol). Then sodium salt of methyl acetoacetate (88.44 g, 0.64 mol) was added in 10 portions with 10 min time interval between portions at -15 °C. The reaction mixture was subsequently allowed toheat up to 20 °C and stirred for an additional 30 min. Then the reaction mixture was cooled to 0 °C, a solution of NaOH (51.0 g, 1.28 mol) in water (150 mL) was added and the reaction mixture was stirred at 50 °C for 2 h. The methanol was removed in vacuo, the residual suspension was diluted with water (1500 mL) and the pH adjusted to 2.0-2.5 with 20% H2SO4 (250 mL). The compound was filtered, washed with water (2x500 mL) and dried in vacuo to give the product as yellow solids. Yield: 129.33 g (85%). LC-MS: MS (ESI) calcd. for: C11H9CINO3 [M+H]+238.02; found: 237.97, tR = 3.60 min. Nuclear magnetic resonance (NMR): (DMSO-rfe, 500 MHz, δ, ppm): 2.72 (s, 3H), 7.42-7.48 (m, 2H), 7.50-7.54 (m, 1H), 7.57-7.59 (m, 1H), 12.98 (br. s, 1H).13CNMR (DMSO-<76, 125 MHz, 8, ppm): 12.85, 109.98, 127.01, 128.43, 129.27, 131.15, 131.26, 132.93, 160.62, 162.30, 175.06. d) Synthesis of 3-(2-chlorophenyl)-5-methyl-N-(6-nitrobenzo[d]thiazol-2-yl)isoxazole-4- carboxamide

[0135] To a solution of 3-(2-chlorophenyl)-5-methylisooxazole-4-carboxylic acid (100 g, 0.42 mol), 2-amino-6-nitrobenzothiazole (90.75 g, 0.46 mol, 1.1 eq) and ethyl cyanohydroxyiminoacetate (Oxyma; 68.25 g, 0.48 mol, 1.1 eq) in DMF (900 mL) were added triethylamine (70.0 mL, 0.50 mol, 1.2 eq) and diisopropylcarbodiimide (DIC) (85.0 mL, 0.55 mol, 1.3 eq). The reaction mixture was stirred at 50 °C under nitrogen atmosphere for 16 h, allowed to cool down to r.t., then kept in a fridge at 5 °C for 2 h. The resulting suspension was filtered, the filter cake was washed with DMF (2x50 ml), and the filtrate was concentrated in vacuo. The residue was dissolved in ethyl acetate (3000 mL), extracted with 2% K2CO3 in water (2x1000 mL), 0.2 M HC1 (2x1000 mL), brine (2x1000 mL), dried over anhydrous sodium sulphate (50 g), filtered and concentrated. The product was dissolved in ethyl acetate (20 mL / g) under reflux and activated charcoal powder (1 g per 10 g of product) was added. The resulting suspension was heated to reflux for 10 minutes under intensive stirring, then filtered by hot filtration. The filtrate was heated to reflux, and heptane (10 mL / g) was added slowly. The solution was allowed to cool down to room temperature and stirred gently overnight. Yield:121.7g (70%). LC-MS: MS (ESI) calcd. for: C18H12CIN4O4S [M+H]+415.02; found: 414.84, tR= 4.26 min. 1HNMR (CDCI3, 500 MHz, δ, ppm): 2.89 (s, 3H), 7.49-7.65 (m, 4H), 7.69 (d, J= 9.0 Hz, 1H), 8.28 (dd, J= 9.0, 2.3 Hz, 1H), 8.73 (d, J = 2.3 Hz, 1H).13C NMR (CDCI3 125 MHz, δ, ppm): 13.72, 110.21, 118.29, 120.92, 122.34, 126.36, 128.32, 131.02, 131.93, 132.42, 133.02, 133.96, 144.35, 152.02, 158.08, 159.46, 161.66, 177.08.II. Discussion

[0136] 3-(2-chlorophenyl)-5-methyl-N-(6-nitrobenzo[d]thiazol-2-yl)isoxazole-4- carboxamide was obtained according to Scheme 2.Scheme 2. i) NH2OH HCI, NaOH, EtOH / water, 20°C, 2h; ii) NCS, DMF, 20°C, 0.5h; iii) sodium salt of methyl acetoacetate, NaOH, MeOH, -20°C to +50°C, 6h; iv) 2-amino-6- nitrobenzothiazole, DIC, Oxyma, EtsN, DMF, 50°C, 16h.

[0137] Briefly, 2-chlorobenzaldehyde oxime was prepared by treatment of 2- chlorobenzaldehyde with hydroxylamine hydrochloride in the presence of sodium hydroxide in ethanol. Then, by chlorination with N-chlorosuccinimide (NCS) in DMF, 2- chlorobenzaldehyde oxime was converted to 2-chloro-N-hydroxybenzimidoyl chloride. Cyclization of 2-chloro-N-hydroxybenzimidoyl chloride with sodium salt of methyl acetoacetate followed by hydrolysis with NaOH gave 3-(2-chlorophenyl)-5-methylisoxazole- 4-carboxylic acid. Finally, amide coupling of 3-(2-chlorophenyl)-5-methylisoxazole-4- carboxylic acid with 2-amino-6-nitrobenzothiazole in the presence of diisopropylcarbodiimide (DIC) and Oxyma yielded 3-(2-chlorophenyl)-5-methyl-N-(6- nitrobenzo[d]thiazol-2-yl)isoxazole-4-carboxamide.Example 2: Biological ActivityI. Materials and Methods

[0138] FA CS-FRET of SWAP- 70 in living cells: The method was in line with that previously described by Betaneli and Jessberger (2020). Briefly, two versions of SWAP-70, differently tagged with the fluorescent dyes Cerulean and Venus, were stably expressed in 293T cells. Upon activation of the cells, e.g., by sodium vanadate, SWAP-70 dimerizes. Thereby the two fluorophores associate and generate a fluorescence resonance energy transfer (FRET) signal, which is measured by FACS. FACS-FRET allows to analyze large numbers of cells simultaneously. The left plot in FIG. 1 shows inhibition of the FACS-FRET signal (FRET efficiency) and thus of SWAP-70 dimerization by, from left to right: 50, 25 and 10 μM B2 in comparison to control (far left), measured at 20 minutes after stimulation of the cells. B2 was added to the cells 15 min prior to stimulation.

[0139] Binding of B2 to SWAP-70: In the microscale thermophoresis (MST) assay the influence of binding of one molecule (B2) to a target molecule (SWAP-70) in solution on the mobility of the target molecule was determined. FIGs. 2-4 show the binding of B2 to the target molecule (SWAP-70) in solution assessed in the microscale thermophoresis (MST) assay. The assay measures the effect of binding of B2 to the target molecule (SWAP-70, fluorescently labeled) on the mobility of the target molecule. FIG. 2 shows the mobility of SWAP-70 at different time points up to 20 minutes after incubation of SWAP-70 with B2; note the change in signal of the complex compared to the target. FIG. 3 shows control measurements of the fluorescence signals at the 14-15 minute interval after incubation of SWAP-70 with B2; and FIG. 4 shows quantification of the mobility shift and thus binding within the same interval, demonstrating significant binding (n = 4).

[0140] Binding of B2 to SWAP- 70: In the nanoDSF (differential scanning fluorimetry) assay, the thermal protein stability based on intrinsic fluorescence of a protein is determined. The lower curve in FIG. 5 represents B2 added to SWAP -70, and the upper curve SWAP-70 only. The two phases represent two melting points (Tml, Tm2).

[0141] Inhibition of tumor cell growth: The inhibition of tumor cell growth of 4T1 mouse breast carcinoma tumor cells was determined using a soft agar colony formation assay. Thisassay tests the ability of tumor cells to form colonies in a 3D matrix in an anchorage- independent manner. 1 ml of 0.5% (w / v) agar in Roswell Park Memorial Institute (RPMI) tissue culture media was added to each well of a 6-well plate and allowed to solidify to create a base. 3 x 104cells were mixed with agarose (final concentration of agarose 0.35% (w / v) in RPMI) and 300 pl of the mixture was layered onto base. The cell culture medium was added on top of the agarose and changed every three days. After two weeks the cells were fixed with 10% methanol and 10% acetic acid in ddFEO for 10 minutes and colonies were stained with 0.01 % crystal violet for 1 hour. Crystal violet was removed and the samples were washed several times with ddH2O until the stained single colonies were clearly visible, then the wells were scanned using an Epson Perfection 4180 scanner and colonies were counted with Fiji software; from left to right in FIG. 6: 0 μM (far left images), 50 μM B2 (second images from left), 20 μM B2 (second images from right) or 10 μM (far right images) was added. FIG. 7 shows the comparison of the number of colonies for the results of three experiments. The control lacking B2 was performed using the solvent for B2, DMSO.

[0142] Inhibition of tumor cell migration: The inhibition of tumor cell migration of 4T1 mouse breast carcinoma cell migration was determined using a membrane in a Transwell set- up. 5 x 104cells in 100 pl low FCS (0.5 %) medium were added to the upper chamber of a Transwell insert (6.5 mm diameter, 8 pm pore size; Coming), and the insert was placed in a 24-well plate containing 1 ml complete medium with transforming growth factor- beta / epidermal growth factor (TGFp / EGF). The Transwell inserts were fixed with 4% paraformaldehyde after 24 hours. Cells that remained on the upper side of the Transwell were removed with a cotton swab, while cells successfully migrated to the bottom of the Transwell were stained with the 4',6-diamidino-2-phenylindole (DAPI) staining solution. Three randomly selected fields on the lower side of the insert were imaged with an Olympus IX 70 inverted microscope at lOx magnification, and the number of cells in each field was then counted using Fiji software. The average number of cells per insert was calculated for 0 μM, 20 μM or 50 μM B2 added. The control lacking B2 was performed using the solvent for B2, DMSO.

[0143] The inhibition of tumor cell migration of 4T1 mouse breast carcinoma cell migration was further determined using a wound healing assay. In this cell culture assay, migration of cells attached to a biological surface into an open space (the “wound”) ismonitored. Flat glass-bottom 96-well plates were pre-coated with fibronectin (bovine plasma fibronectin, Merck Sigma- Aldrich) which had been resuspended in phosphate-buffered saline (PBS) with 1 mM MgCI2 and 1 mM CaCI2 at a concentration of 25 pg / ml for 2 hours at 37°C. The Oris™ (Platypus technologies) silicon stopper was placed into the 96-well fibronectin- coated plate to create a physical barrier, and 5 x 104cells were seeded around the stopper with complete cell medium with, or without, TGFp / EGF overnight at 37°C. After removing the stoppers, migration of the cells into the resulting void was imaged for each sample every 30 minutes for up to 15 hours on a Nikon TE2000E inverted microscope with a Plan APO 4x / 0.13 NA objective, a CoolSNAP HQ camera (Photometries), and standard filter sets at 37°C. All images were acquired using the Nikon NIS-Elements ND2 software and analyzed with Fiji. The control lacking B2 was performed using the solvent for B2, DMSO.

[0144] Inhibition of 3D spheroid formation: The inhibition of 3D spheroid formation was determined by culturing a single cell suspension of 4T1 cells in ultra-low-attachment plates and measuring formation of cell aggregates, spheroids, using microscopy or Alamar blue staining. B2 was added at the onset of the cultures in different concentrations, i.e. no B2 (DMSO), 50 μM, 10 μM, 1 μM and 0.1 μM.

[0145] Inhibition of tumor growth in mice: Balb / c mice (10 mice each group; 25 g body weight, females) were orthotopically injected into to mammary fat pad with 1 x 1064T1 mouse breast cancer cells. Starting at day 3 after injection of the tumor cells, every 3 days the mice received an injection into the tail vein of 50 pl of a 2.5 mg / ml solution of B2, calculated to result in an initial serum concentration of 142 μM B2 in 1% DMSO / PBS or, for control, 1% DMSO / PBS only. The mice were euthanized after a total of 21 days. Tumor volume and weight were measured.IE Results

[0146] B2 inhibits SWAP-70 dimerization in living cells (FIG. 1). Two versions of SWAP-70, differently tagged with the fluorescent dyes Cerulean and Venus, were stably expressed in 293T cells. Upon activation of the cells, e.g., by sodium vanadate, SWAP-70 dimerizes. Thereby the two fluorophores associate and generate a fluorescence resonance energy transfer (FRET) signal, which is measured by FACS. FACS-FRET allows to analyze large numbers of cells simultaneously. The left plot in FIG. 1 shows inhibition of the FACS-FRET signal (FRET efficiency) and thus of SWAP-70 dimerization by, from left to right: 50 μM, 25 μM and 10 μM B2 in comparison to control (far left), measured at 20 minutes after stimulation of the cells. The right plot in FIG. 1 shows inhibition by 50 μM B2 at earlier time intervals (second column from left: 5-10 minutes; far right: 10-15 minutes) in comparison to controls (far left: 5-10 minutes; second column from right: 10-15 minutes) after stimulation. B2 was added to the cells 15 minutes prior to stimulation.

[0147] To investigate the binding of B2 to SWAP-70 the mobility of the target molecule in solution was determined using the microscale thermophoresis assay (MST) at different time points up to 20 minutes and 2 h after incubation of the target with B2 (FIGs. 2-4). The upwards shift of the right curve (complex of SWAP-70 and B2) demonstrates binding; left curve: SWAP- 70 only (FIG. 3). Similar results obtained after 2 h incubation show stability of complex formation for at least 2 h (FIG. 4).

[0148] To independently investigate the binding of B2 to SWAP-70 the thermal protein stability was determined using the nanoDSF (differential scanning fluorimetry) assay, based on intrinsic fluorescence of the protein (FIG. 5), wherein B2 was added to SWAP-70, and compared to SWAP-70 only. The lower curve representing B2 added to SWAP -70 differs from the upper curve (SWAP-70 only), indicating binding of B2 to the protein. The two phases representing two melting points (Tml, Tm2) are shifted upwards. Binding of B2 renders SWAP -70 more resistant to heat-denaturation as indicated by the higher melting temperature (Tml of 49.1 °C and Tm2 of 58.0°C) in comparison to SWAP-70 only (Tml of 44.9°C and Tm2 of 53.4°C). FIGs. 2-4 and FIG. 3 show direct binding of B2 to SWAP-70 in two separate, independent assays.

[0149] To investigate the inhibition of tumor cell growth 4T1 mouse breast carcinoma, tumor cells were treated with B2 in a soft agar colony formation assay. 4T1 mouse breast carcinoma tumor cells were treated with 0 μM, 10 μM, 20 μM or 50 μM B2 and fixed and stained after two weeks (FIG. 6). The resulting number of colonies are shown in FIG. 7. Inhibition of tumor cell growth was observed at concentrations of 10 μM, 20 μM and 50 μM B2.

[0150] To investigate the inhibition of tumor cell migration 4T1, mouse breast carcinoma cells were treated with B2 in a membrane in a Transwell set-up. 4T1 mouse breast carcinoma tumor cells were treated with 0 μM, 10 μM or 50 μM B2 and fixed in the bottom of the Transwell after 24 hours and imaged with an Olympus IX 70 inverted microscope at lOxmagnification. The average number of cells per insert is shown in FIG. 8. Inhibition of tumor cell migration was observed at a concentration of 50 μM B2. FIG. 9 shows images showing representative fields of view of migrated cells for 0 μM (left image) and 50 μM B2 (right image).

[0151] To further investigate the inhibition of tumor cell migration 4T1, mouse breast carcinoma cells were treated with B2 in a wound healing assay. 4T1 mouse breast carcinoma tumor cells were treated with 0 μM, 10 μM, 20 μM or 50 μM B2 and were imaged for each sample every 30 minutes for up to 15 hours on a Nikon TE2000E inverted microscope with a Plan APO 4x / 0.13 NA objective, a CoolSNAP HQ camera (Photometries), and standard filter sets at 37°C. The percentage of “wound” area covered by 4T1 breast carcinoma tumor cells 18 hours after addition of three concentrations of B2 (10 μM, 20 μM or 50 μM) and the control (DMSO) is shown in FIG. 10. Significant inhibition of tumor cell migration was observed at a concentration of 50 μM B2. The mobility of control-treated cells was much higher than those treated with 50 μM B2. FIG. 11 shows exemplary images showing a “wound” within a cell layer in the wound healing assay. The white arrow points to the edge of the wound where cells are actively migrating into the free space only in the control (DMSO, left image).

[0152] The inhibition of 3D spheroid formation was determined by culturing a single cell suspension of 4T1 cells in ultra-low-attachment plates and measuring formation of cell aggregates, spheroids, using microscopy or Alamar blue staining (FIGs. 12 and 13).The inhibition of tumor growth in mice was examined by injecting 4T1 cells into to mammary fat pad of mice, injecting B2 or DMSO / PBS (control) after 3 days and measuring tumor volume and tumor weight (FIG. 14). The mice were euthanized after a total of 21 days or if the tumor became necrotic with break-through through the skin. None of the mice had to be euthanized before 21 days. Upon treatment with B2, the reduction in tumor volume was 22% and the reduction in tumor weight 27%. Furthermore, there was 21% more necrosis in the B2-treated tumor. III. Discussion

[0153] The compound B2 was selected in vitro for inhibition of SWAP-70 and subsequently shown to block SWAP-70 dimerization, which is key to its activity (FIG. 1).

[0154] To show direct target engagement, SWAP-70 was incubated with B2 and the change in mobility caused by binding measured in the MST assay (FIGs. 2-4). The complex showed altered MST trace curves proving binding of B2 to the protein.

[0155] To show direct target engagement in a second, independent assay, SWAP-70 was incubated with B2 and subjected to thermal treatment in a nanoDSF assay. The denaturation of the protein, indicated by change of its intrinsic fluorescence, was altered by B2, demonstrating direct association of B2 to SWAP-70 (FIG. 5).

[0156] Anchorage-independent growth of tumor cells, a hallmark of cancer, is strongly reduced by treatment of tumor cells with B2 (FIGs. 6 and 7).

[0157] Tumor cell migration is a key feature of cancer, particularly for metastasis. FIGs. 8 and 9 show inhibition of tumor cell migration in vitro by B2.

[0158] Expanding into free space in cell culture by migration and proliferation is another property of tumor cells and measured as “wound healing”, i.e. filling of empty space within a monolayer of cells. This activity is inhibited by B2 (FIGs. 10 and 11).

[0159] F ormation of spheroids by tumor cells is another hallmark of tumor cells and is inhibited by B2 as shown in FIGs. 12 and 13. FIG. 12 shows quantification of spheroids by area (left) and fluorescence (right) as the tumor cells were equipped with a fluorescent dye. FIG. 13 shows examples of spheroids untreated (DMSO) or treated with the indicated concentrations of B2.

[0160] While the present disclosure has been described with reference to examples, it is to be understood that the scope of the claims should not be limited by the embodiments set forth in the examples but should be given the broadest interpretation consistent with the description as a whole.

[0161] All publications, patents and patent applications arc herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety. Where a term in the present application is found to be defined differently in a document incorporated herein by reference, the definition provided herein is to serve as the definition for the term.FULL CITATIONS FOR REFERENCES REFERRED TO IN THE DESCRIPTIONAdams, J. C. (2004). "Roles of fascin in cell adhesion and motility." Curr Opin Cell Biol 16(5): 590-596.Barik, G. K., O. Sahay, D. Paul and M. K. Santra (2022). "Ezrin gone rogue in cancer progression and metastasis: An enticing therapeutic target." Biochim Biophys Acta Rev Cancer 1877(4): 188753.Betaneli, V. and R. Jessberger (2020). "Mechanism of control of F-actin cortex architecture by SWAP-70." J Cell Sci 133(2).Borggrefe, T., M. Wabl, A. T. 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Claims

CLAIMSWhat is claimed is:

1. A compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof for use in treating cancer:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is an integer of from 0 to 4.

2. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of claim 1, wherein the cancer is selected from prostate cancer, breast cancer, cervical cancer, thyroid cancer, colorectal cancer, stomach cancer, liver cancer, lung cancer, lymphoma, leukemia, glioblastoma, sarcoma and melanoma.

3. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of claim 1 or 2, wherein the cancer comprises cancer stem cells.

4. A compound of Formula I or a pharmaceutically acceptable salt, solvate and / or prodrug thereof for use in reducing and / or preventing tumor invasion and / or metastasis:whereinR1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2and R3are each independently selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2and SO2R5;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is an integer of from 0 to 5; and n is an integer of from 0 to 4.

5. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of claim 4, wherein the tumor invasion and / or metastasis is in a subject having a primary cancer selected from prostate cancer, breast cancer,cervical cancer, thyroid cancer, colorectal cancer, stomach cancer, liver cancer, lung cancer, glioblastoma, sarcoma and melanoma.

6. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of claim 5 or 6, wherein the primary cancer comprises cancer stem cells.

7. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 6, wherein X2is O.

8. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 7, wherein m is 1.

9. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 8, wherein n is 1.

10. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 6, wherein:R1is selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl;R2is selected from C1-10alkyl, substituted or unsubstituted aryl, heteroaryl, halo, OR4, NO2 and SO2R5;R3is NO2;X1is selected from N and CH;X2is selected from O, S and NR6;X3is selected from O and NR7;X4is selected from O, S, CH2, SO, SO2 and NR8;X5is selected from N and CH;R4, R5, R6, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; m is 1 ; and n is an integer of from 1 to 4.

11. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of claim 10, wherein:R4, R5, R7and R8are each independently selected from H, C1-10alkyl, substituted or unsubstituted aryl and heteroaryl; andR6is selected from H and C1-10alkyl.

12. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 6, wherein the compound of Formula I is a compound of Formula 1(a):wherein R1, R2, R3, X1, X2, X3, X4and X5are as defined in any one of claims 1, 4, 7, 10 or 11.

13. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of claim 1 or 2, wherein the compound of Formula I is a compound of Formula I(a)(i):wherein R1, R2, R3, X1, X3, X4and X5are as defined in any one of claims 1, 4 or 10.

14. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of claim 12 or 13, wherein R3is NO2.

15. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 14, wherein R1is Ci-4alkyl.

16. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of claim 15, wherein R1is methyl.

17. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 16, wherein R2is halo.

18. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of claim 17, wherein R2is chloro.

19. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 18, wherein X1is N.

20. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 19, wherein X3is NH.

21. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 20, wherein X4is S.

22. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 21, wherein X5is N.

23. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 6, wherein the compound of Formula I has the structure:

24. The compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for the use of any one of claims 1 to 23, wherein the compound of Formula I or the pharmaceutically acceptable salt, solvate and / or prodrug thereof is the compound of Formula I.

25. A pharmaceutical composition comprising a compound as defined in any one of claims 1 to 23 or the pharmaceutically acceptable salt, solvate and / or prodrug thereof for use in treating cancer.

26. The pharmaceutical composition for the use of claim 25, further comprising a pharmaceutically acceptable carrier.

27. The pharmaceutical composition for the use of claim 26, wherein the cancer is selected from prostate cancer, breast cancer, cervical cancer, thyroid cancer, colorectal cancer, stomach cancer, liver cancer, lung cancer, lymphoma, leukemia, glioblastoma, sarcoma and melanoma.

28. The pharmaceutical composition for the use of any one of claims 25 to 27, wherein the cancer comprises cancer stem cells.