2-hydroxypropyl-beta-cyclodextrin as neoadjuvant or adjuvant for cancer therapy

Abstract

The present invention relates to a compound, 2-hydroxypropyl-beta-cyclodextrin (HP [beta] CD), for use in a method of treating cancer, a method of treating cancer using the compound HP [beta] CD, and the use of the compound in the preparation of one or more medicaments for the treatment of cancer. In particular, the compound HP [beta] CD can be used as adjuvant therapy or neoadjuvant therapy. Also provided are pharmaceutical compositions comprising HP [beta] CD, optionally comprising other chemotherapeutic agents or cancer therapeutic agents.

Description

Background Art

[0001] The present invention relates to a compound for treating cancer, namely 2-hydroxypropyl-β-cyclodextrin (HPβCD), a method for treating cancer using the compound HPβCD, and the use of the compound in preparing one or more drugs for treating cancer. Also provided is a pharmaceutical composition containing HPβCD, which optionally contains other chemotherapeutic drugs or cancer therapeutic agents.

[0002] Cancer is one of the leading causes of death worldwide. According to the World Health Organization (WHO), new cancer cases are expected to increase by about 70% in the next 20 years. Breast cancer is one of the most common malignancies among women worldwide, with nearly 1.7 million new cases worldwide in 2012. In 2012, there were more than 8,000 new cases of breast cancer in South Africa, accounting for 21.79% of the total number of cancer diagnoses in the country (National Health Laboratory Service, 2012).

[0003] Currently, cancer treatments include chemotherapy and, when possible, surgical removal of the tumor. Even when treatment is successful, cancer metastasis and recurrence remain the main reasons for low patient survival. Epithelial-mesenchymal transition (EMT) is a cellular program by which tumor cells acquire metastatic capacity and drug resistance. Existing breast cancer treatments include the use of selective estrogen receptor modulators (SERMs), a class of drugs that includes tamoxifen (the gold standard drug), but are known to have a variety of side effects. These side effects include uterine cancer, cataracts, thrombosis, and heart attacks. Despite significant advances in anticancer therapy, many cancer patients develop resistance to drug treatments, either innate or acquired, and often experience disease recurrence. Notably, invasion and metastasis mediated by malignant cancer cells account for 90% of cancer patient deaths. Therefore, it is critical to develop new approaches to inhibit cancer metastasis and address drug resistance.

[0004] Emerging experimental evidence suggests that imbalanced cholesterol homeostasis plays a key role in promoting cancer development and drug resistance. In addition, tumor metastasis often favors the transition between epithelial and mesenchymal cell states. Epithelial-mesenchymal transition (EMT) has been reported to lead to drug resistance, allowing cancer to relapse and metastasize after chemotherapy. High cholesterol levels not only directly regulate protein intermediates but also promote increased formation of lipid rafts, which are important signaling centers that are essential for the induction of EMT. Cholesterol is co-enriched with sphingolipids in specific areas of the cell membrane to form microdomains called lipid rafts. Lipid rafts are known to play a central role in multiple key signaling pathways involved in cell survival, migration, cell cycle progression, and tumorigenesis and metastasis. Studies have shown that during cells undergoing epithelial-mesenchymal transition (EMT), cholesterol levels increase significantly, while ceramide pools decrease, thereby enhancing plasma membrane fluidity. Stabilization of lipid rafts has been proposed as an emerging therapeutic target to reduce the increased metastatic potential mediated by epithelial-mesenchymal transition. This view is supported by studies showing that cancer cells that have undergone epithelial-mesenchymal transition are more sensitive to cholesterol-targeting drugs (i.e., statins), which reduce the cholesterol content in cells and, by reducing membrane fluidity, reduce cell motility and the potential for metastasis.

[0005] Since the induction of EMT is associated with an increased likelihood of drug resistance, it is plausible that cholesterol contributes to this increased likelihood of drug resistance through its EMT-regulating effects. Furthermore, dysregulated cholesterol metabolism is also thought to confer chemoresistant properties to cells, leading to drug resistance. This process is achieved through cholesterol-mediated regulation of pro-survival and anti-apoptotic signaling pathways. Furthermore, given the critical role of cholesterol in maintaining lipid raft integrity, there is evidence that lipid rafts are key domains for maintaining the activity of multidrug resistance transporters. Therefore, this points to multiple roles for cholesterol in promoting EMT and mediating cancer drug resistance.

[0006] Currently, the strategy of targeting cellular cholesterol by inhibiting cholesterol synthesis to restore sensitivity has been widely used in laboratory settings. This is mainly achieved by using the most commonly used lipid-lowering drugs on the market, namely statins. Although inhibiting cholesterol synthesis to reduce cellular cholesterol levels and enhance drug sensitivity has been shown to have certain effects in laboratory settings, the debilitating side effects associated with statin therapy are considered to be its main limiting factor in clinical settings. Common side effects include but are not limited to muscle pain, hepatotoxicity and nephrotoxicity, neurocognitive decline, myopathy, and idiopathic polyneuropathy. Notably, 30% of patients do not respond to statin treatment. This phenomenon may be attributed to the long-term inhibition of cholesterol synthesis in cells, where cholesterol plays a key role in regulating basic cellular processes. Therefore, the discovery of new cholesterol-lowering drugs is crucial to overcome resistance.

[0007] The present inventors propose that targeting cellular cholesterol may be a promising area for finding new cancer treatments and have identified a compound that may be used as an adjuvant or neoadjuvant therapy. Summary of the invention

[0008] The present invention relates to a compound for treating cancer, namely 2-hydroxypropyl-β-cyclodextrin (HPβCD), a method for treating cancer using the compound HPβCD, and the use of the compound in preparing one or more drugs for treating cancer. In addition, according to the present invention, a pharmaceutical composition comprising HPβCD is also provided, and the pharmaceutical composition optionally comprises other chemotherapeutic drugs.

[0009] According to a first aspect of the present invention, there is provided 2-hydroxypropyl-β-cyclodextrin (HPβCD) for use in a method of treating cancer, the method comprising administering HPβCD in combination or co-administration with another cancer therapy to a subject in need thereof.

[0010] In a first embodiment of use of HPβCD, HPβCD may be administered as a neoadjuvant prior to another cancer therapy.

[0011] According to a second embodiment of the use of HPβCD, when HPβCD is administered as a neoadjuvant, administration of HPβCD increases the sensitivity of cancer cells to cancer therapy by reversing the epithelial-mesenchymal transition phenotype of cancer cells.

[0012] In a third embodiment of the use of HPβCD, administering HPβCD as a neoadjuvant agent can improve a subject's response to cancer therapy.

[0013] According to a fourth embodiment of the use of HPβCD, HPβCD may be administered as an adjuvant in combination with or after another cancer therapy.

[0014] In another embodiment of the use of HPβCD, HPβCD is administered as an adjuvant to sensitize non-proliferative or proliferative cancer cells to cancer therapy.

[0015] According to another embodiment of the use of HPβCD, administration of HPβCD can reduce the toxicity of cancer therapy and / or reduce one or more side effects of cancer therapy. For example, a subject may exhibit better tolerance to cancer therapy (such as oxaliplatin therapy or fluorouracil therapy) when used in combination with HPβCD.

[0016] According to another embodiment of the use of HPβCD, administering HPβCD as an adjuvant can reduce the risk or incidence of cancer recurrence.

[0017] In another embodiment of HPβCD use, HPβCD may be combined or co-administered with another cancer therapy selected from the group consisting of chemotherapy, radiation therapy, hormone therapy, targeted therapy, or biological therapy.

[0018] According to a second aspect of the present invention, there is provided a method of treating cancer, the method comprising administering 2-hydroxypropyl-β-cyclodextrin (HPβCD) in combination or co-administering with another cancer therapy to a subject in need thereof.

[0019] In a first embodiment of this method, HPβCD may be administered as a neoadjuvant prior to another cancer therapy.

[0020] According to a second embodiment of the method, when HPβCD is administered as a neoadjuvant, administration of HPβCD can increase the sensitivity of cancer cells to cancer therapy by reversing the epithelial-mesenchymal transition phenotype of cancer cells.

[0021] In a third embodiment of this method, administering HPβCD as a neoadjuvant improves a subject's response to cancer therapy.

[0022] According to a fourth embodiment of this method, HPβCD may be administered as an adjunct agent in conjunction with or after another cancer therapy.

[0023] In another embodiment of this method, HP[beta]CD is administered as an adjunct agent to sensitize non-proliferative or proliferative cancer cells to cancer therapy.

[0024] According to another embodiment of the method, administering HPβCD as an adjunct agent reduces the risk or incidence of cancer recurrence.

[0025] According to another embodiment of the method, administering HPβCD can reduce the toxicity of a cancer therapy and / or reduce one or more side effects of a cancer therapy. For example, a subject may exhibit better tolerance to a cancer therapy (such as oxaliplatin therapy or fluorouracil therapy) when administered in combination with HPβCD.

[0026] In another embodiment of this method, HPβCD may be combined or administered in conjunction with another cancer therapy selected from the group consisting of chemotherapy, radiation therapy, hormone therapy, targeted therapy, or biological therapy.

[0027] According to a third aspect of the present invention, there is provided a use of 2-hydroxypropyl-β-cyclodextrin (HPβCD) in the preparation of a medicament for a method for treating cancer, the method comprising combining the medicament with another cancer therapy or co-administering the medicament to a subject in need thereof. It will be appreciated by those skilled in the art that when the other cancer therapy is a chemotherapeutic drug, a biological agent, a hormone or a targeted therapy drug, the medicament may be co-formulated with the cancer therapy or separately formulated.

[0028] In a first embodiment of this use, the medicament comprising HPβCD may be administered as a neoadjuvant prior to another cancer therapy.

[0029] According to a second embodiment of the use, when the drug comprising HPβCD is administered as a neoadjuvant, the administration of the drug can increase the sensitivity of cancer cells to cancer therapy by reversing the epithelial-mesenchymal transition phenotype of cancer cells.

[0030] In a third embodiment of this use, administering a medicament comprising HPβCD as a neoadjuvant improves a subject's response to cancer therapy.

[0031] According to a fourth embodiment of the use, the medicament comprising HPβCD can be administered as an adjuvant in combination with another cancer therapy or after another cancer therapy.

[0032] In another embodiment of this use, administration of a medicament comprising HP[beta]CD as an adjunct agent can sensitize non-proliferative or proliferative cancer cells to cancer therapy.

[0033] According to another embodiment of the use, administering a drug comprising HPβCD can reduce the toxicity of cancer therapy and / or reduce one or more side effects of cancer therapy. For example, when combined with a drug comprising HPβCD, a subject may exhibit better tolerance to cancer therapy (such as oxaliplatin therapy or fluorouracil therapy).

[0034] According to another embodiment of the use, administering a medicament comprising HPβCD as an adjuvant can reduce the risk or incidence of cancer recurrence.

[0035] In another embodiment of this use, the medicament comprising HPβCD may be combined or co-administered with another cancer therapy selected from the group consisting of chemotherapy, radiotherapy, hormone therapy, targeted therapy, or biological therapy.

[0036] According to a fourth aspect of the present invention, there is provided a pharmaceutical composition comprising 2-hydroxypropyl-β-cyclodextrin (HPβCD) and a pharmaceutically acceptable carrier, and optionally comprising a chemotherapeutic drug. It will be appreciated by those skilled in the art that the pharmaceutical composition can be formulated together with the chemotherapeutic drug or separately.

[0037] In one embodiment, the pharmaceutical composition may further comprise a pharmaceutically acceptable excipient, a diluent, a carrier or other suitable additives. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Non-limiting embodiments of the present invention will be described, by way of example only, and with reference to the following drawings.

[0039] Figure 1 : Effects of CoCl2, TGF-β, and IL-6 treatment on E-cadherin, an epithelial marker for epithelial-mesenchymal transition. Protein expression of E-cadherin markers in control and CoCl2, TGF-β, and IL-6-treated cells was detected by immunofluorescence microscopy. E-cadherin expression was indicated by Alexa Fluor 488 staining (green), and nuclei were stained with DAPI (blue). The scale bar in each image represents 100 μM. Statistical analysis was performed using an unpaired two-tailed t-test, and data are expressed as mean ± SD, n = 3. * P<0.05, ** P<0.01, *** P<0.001, ns indicates no statistical difference.

[0040] Figure 2 : Effects of CoCl2, TGF-β, and IL-6 treatment on vimentin, a mesenchymal marker of epithelial-mesenchymal transition. Protein expression of epithelial-mesenchymal transition molecular markers in the control group and cells treated with CoCl2, TGF-β, and IL-6 was detected by immunofluorescence microscopy. Vimentin expression was indicated by Texas Red staining (red), and cell nuclei were stained with DAPI (blue). The scale bar of each image represents 100 μM. Statistical analysis was performed using an unpaired two-tailed t-test, and data are expressed as mean ± SD, n = 3. * P<0.05, ** P<0.01, *** P<0.001, ns indicates no statistical difference.

[0041] Figure 3: Quantitative analysis of E-cadherin expression in cells induced by epithelial-mesenchymal transition after cholesterol-targeted drug treatment. Images show the protein expression of E-cadherin after treatment with different cholesterol-targeted drugs (1 mM MβCD and 10 mM HPβCD) before and after epithelial-mesenchymal transition in: (a) MCF-7 cells; (b) NMuMg cells; (c) HT-29 cells. E-cadherin expression was indicated by Alexa Fluor 488 staining (green), and cell nuclei were stained with DAPI (blue). The scale bar in each image represents 100 μM. Statistical analysis was performed using one-way analysis of variance (ANOVA) with Bonferroni post hoc test, and data are presented as mean ± SD, n = 3. * P<0.05, ** P<0.01, *** P<0.001, ns indicates no statistical difference.

[0042] Figure 4 : Quantitative analysis of vimentin expression in cells induced by epithelial-mesenchymal transition after cholesterol-targeted drug treatment. Images show the protein expression of vimentin after treatment with different cholesterol-targeted drugs (1 mM MβCD and 10 mM HPβCD) before and after epithelial-mesenchymal transition in: (a) MCF-7 cells; (b) NMuMg cells; (c) HT-29 cells. Vimentin expression was indicated by Texas Red staining (red), and nuclei were stained with DAPI (blue). The scale bar in each image represents 100 μM. Statistical analysis was performed using one-way analysis of variance (ANOVA) with Bonferroni post hoc test, and data are presented as mean ± SD, n = 3. * P<0.05, ** P<0.01, *** P<0.001, ns indicates no statistical difference.

[0043] Figure 5: Evaluation of the effect of cholesterol depletion on the expression of genes related to epithelial-mesenchymal transition in NMuMg cells. a) RT-qPCR was used to analyze the transcriptional expression levels of CDH1, Vim, SNAI1, TWIST1, ZEB1, CTBP1 and SMAD4 genes in TGFβ-treated cells (after epithelial-mesenchymal transition) relative to cells before epithelial-mesenchymal transition. b) RT-qPCR was used to analyze the transcriptional expression levels of CDH1, Vim, SNAI1, TWIST1, ZEB1, CTBP1 and SMAD4 genes in untreated cells before epithelial-mesenchymal transition and cells after epithelial-mesenchymal transition treated with 1mM MβCD, 10mM HPβCD and 10μM simvastatin for 2 hours. Relative expression levels are expressed as log2 fold changes between conditions. Data are expressed as mean ± SD. Statistical significance was calculated using a two-tailed unpaired Student's t-test, where * P<0.05, ** P<0.01, *** P < 0.001, ns indicates no statistical difference, n = 3. c) Western blot analysis of the protein expression levels of E-cadherin, vimentin, and N-cadherin in cells before and after epithelial-mesenchymal transition, cells were treated with 1 mM MβCD, 10 mM HPβCD, and 10 μM simvastatin for 2 h, and β-tubulin was used as a loading control.

[0044] Figure 6 : Evaluation of the invasion potential of cells induced by epithelial-mesenchymal transition after treatment with cholesterol-targeted drugs. Images represent the number of invasive cells treated with different cholesterol-targeted drugs (1 mM MβCD and 10 mM HPβCD) before and after epithelial-mesenchymal transition induction, including: (a) MCF-7 cells; (b) NMuMg cells; (c) HT-29 cells. Cell nuclei were stained with DAPI (blue) to assist visualization. The scale bar in each image represents 100 μM. Statistical analysis was performed using one-way analysis of variance (ANOVA) with Bonferroni post hoc test, and data are expressed as mean ± SD, n = 3. * P<0.05, ** P<0.01, *** P<0.001, ns indicates no statistical difference.

[0045] Figure 7: Evaluation of the effect of cholesterol depletion on cholesterol-related gene expression in NMuMG cells. a) RT-qPCR was used to analyze the transcriptional expression levels of ABCA1, ABCG1, SREBF2, LXR, LDLR, HMGCR, PCSK9 and LCAT genes in TGFβ-treated cells (after EMT) relative to cells before EMT. b) RT-qPCR was used to analyze the transcriptional expression levels of ABCA1, ABCG1, SREBF2, LXR, LDLR, HMGCR, PCSK9 and LCAT genes in untreated cells before EMT and cells after EMT treated with 1mM MβCD, 10mM HPβCD and 10μM simvastatin for 2 hours. Relative expression levels are expressed as log2 fold changes between conditions. Data are expressed as mean ± SD. Statistical significance was calculated using a two-tailed unpaired Student's t-test, where * P<0.05, ** P<0.01, *** P < 0.001, ns indicates no statistical difference, n = 3. c) Western blot analysis of the protein expression levels of ABCG1, HMGCR, LXR, SREBP1 and SREBP2 (N: nucleus, C: cytoplasm) in cells before and after epithelial-mesenchymal transition. Cells were treated with 1 mM MβCD, 10 mM HPβCD and 10 μM simvastatin for 2 h, and β-tubulin was used as a loading control.

[0046] Figure 8 : Evaluation of the effect of targeted cholesterol on MDR1 transporter function. a) After epithelial-mesenchymal transition, intracellular retention of fluorescent calcein was significantly increased in (a) MCF-7 cells and (b) HT-29 cells treated with cholesterol-depleting drugs. b) HT-29 cell data show that cholesterol-depleting drugs (1 mM MβCD and 10 mM HPβCD) increased intracellular retention of fluorescent calcein in cells treated with FOLFOX chemotherapy regimen. Statistical analysis was performed using one-way analysis of variance (ANOVA) with Bonferroni post hoc test, and data are expressed as mean ± SD, n = 3. * P<0.05, ** P<0.01, *** P<0.001, ns indicates no statistical difference.

[0047] Fig. 9: To investigate the effect of HPβCD combined with colorectal cancer (CRC) chemotherapeutic drugs on tumor size in vivo. NOD / SCID mice were treated with the following single drugs: 3000mg / kg HPβCD, 10mg / kg oxaliplatin (OXAL), and 30mg / kg 5-fluorouracil (5FU), as well as HPβCD+OXAL and HPβCD+5FU combined treatments. The results showed that tumor size was reduced after HPβCD treatment, and tumor size was further reduced after combined treatment.

[0048] Fig.10 : Graphical representation of tumor size and weight in mice after HPβCD combination therapy. Comparison of tumor size in untreated NOD / SCID mice and treated mice (3000 mg / kg HPβCD, 10 mg / kg OXAL and 30 mg / kg 5FU) (upper panel). Comparison of tumor weight in untreated mice and treated mice (lower panel). Combination treatment with HPβCD+OXAL or HPβCD+5FU significantly reduced tumor size and tumor weight compared to the use of these chemotherapeutic drugs alone. Statistical analysis was performed using one-way analysis of variance (ANOVA) with Dunnett's post hoc test, and data are expressed as mean ± SD, n = 4. * P<0.05, ** P<0.01, *** P<0.001, ns indicates no statistical difference.

[0049] Fig.11 : Graphical representation of the average weekly body weight change of mice over 15 weeks.

[0050] Fig.12 : Graphical representation of serum cholesterol levels in NOD / SCID mice after treatment. The mean serum cholesterol level in mice treated with HPβCD alone was slightly higher (65.5 mg / dL) compared to the mean level in untreated mice (51 mg / dL). Statistical analysis was performed using one-way analysis of variance (ANOVA) with Dunnett's post hoc test, and data are presented as mean ± SD, n = 4. There were no significant differences in serum cholesterol levels between all treatment groups.

[0051] Fig.13 : Graphical representation of AST and ALT levels, indicators of hepatotoxicity, after treatment in NOD / SCID mice. ALT measurements were similar between the untreated group and all treated groups (upper panel). An AST:ALT ratio (U / L) greater than 2 indicated hepatotoxicity, which was only observed in the HPβCD+OXAL treated group (lower panel). Statistical analysis was performed using one-way analysis of variance (ANOVA) with Dunnett's post hoc test, and data are expressed as mean ± SD, n = 4. There were no significant differences in toxicity between all treatment groups. DETAILED DESCRIPTION

[0052] The present invention will now be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all embodiments of the invention are shown.

[0053] The description of the present invention should not be limited to the specific embodiments disclosed, and modifications and other embodiments are intended to be included within the scope of the present invention. Although specific terms are used herein, they are used only for generic and descriptive purposes and not for limitation.

[0054] In this specification and the following claims, the singular forms "a," "an," and "the" include plural forms unless the context clearly dictates otherwise.

[0055] The terms and expressions used herein are for descriptive purposes and should not be construed as limiting in any way. The terms "comprising," "containing," "having," and "including," and variations thereof, as used herein, are intended to encompass the items listed thereafter and equivalents thereof, as well as additional items.

[0056] The inventors of the present invention discovered the role of the cholesterol depleting agent HPβCD in the cell state after epithelial-mesenchymal transition and found that it can reverse the mesenchymal phenotype to an epithelial phenotype. The inventors tested this concept in three cell lines, using three different epithelial-mesenchymal transition induction methods, and found similar results in all cell lines. This shows that HPβCD can reverse the mesenchymal phenotype to an epithelial phenotype in different types of cancer, regardless of the specific epithelial-mesenchymal transition induction mediator.

[0057] Based on the experimental results described herein, the inventors have demonstrated that targeting cellular cholesterol through cholesterol depletion can inhibit epithelial-mesenchymal transition, reduce the invasion and metastasis potential of cells, and restore their sensitivity to conventional chemotherapeutic drugs. In addition, the inventors have further demonstrated through in vivo studies in mice that the administration of HPβCD can reduce the toxicity of cancer therapies and can therefore be used to reduce one or more side effects of cancer therapies, such as oxaliplatin therapy or fluorouracil therapy, while preventing cancer metastasis.

[0058] Therefore, the inventors propose that the use of HPβCD in combination with conventional cancer therapies in neoadjuvant and adjuvant settings could improve patient response to treatment and reduce relapse rates. In addition, the reduction in drug resistance observed with HPβCD treatment could allow for a reduction in the dose of chemotherapy drugs, thereby reducing the severe side effects often associated with high-dose chemotherapy.

[0059] As mentioned above, HPβCD has potential application value in neoadjuvant and adjuvant therapy. Neoadjuvant therapy includes chemotherapy, radiotherapy or hormone therapy, which is usually performed before surgical resection of the tumor. Combining HPβCD with existing neoadjuvant therapy can improve the sensitivity of tumors to drugs by reversing the epithelial-mesenchymal transition phenotype, thereby enhancing the therapeutic effect.

[0060] HPβCD can be used as an adjuvant therapy in combination with existing therapies to reduce the risk of recurrence. Existing treatments, such as chemotherapy and radiation, target proliferative cells. Recurrence occurs if non-proliferative tumor cells remain after treatment. Adding HPβCD to the adjuvant therapy regimen can sensitize non-proliferative cells and / or proliferative cells to existing therapies, thereby reducing the risk of recurrence.

[0061] Therefore, it is proposed that HPβCD can be used in combination with existing chemotherapy drugs to reduce their toxicity, improve efficacy, prevent metastasis and reduce tumor resistance. In addition, HPβCD also has the potential benefit as a long-term treatment drug to prevent cancer recurrence or spread. As shown in the following examples, HPβCD has the potential to reverse the mesenchymal phenotype to the epithelial phenotype, thus indicating its ability to prevent cancer spread.

[0062] The pharmaceutical compositions and compounds of the present invention can be provided alone or in combination with other compounds (e.g., nucleic acid molecules, small molecules, peptides or peptide analogs), in the presence of liposomes, adjuvants or any carrier (e.g., a pharmaceutically acceptable carrier), and in a form suitable for administration to mammals (e.g., humans).

[0063] As used herein, "pharmaceutically acceptable carriers" or "excipients" include any or all physiologically compatible substances such as antibacterial and antifungal agents, coatings, dispersion media, solvents, isotonic agents, and retarder agents. "Pharmaceutically acceptable carriers" may include solid or liquid fillers, diluents, or encapsulating materials that can be safely used to administer a pharmaceutical composition or compound to a subject. The pharmaceutically acceptable carrier may be suitable for intramuscular, intradermal, intravenous, intraperitoneal, subcutaneous, oral, or sublingual administration. Pharmaceutically acceptable carriers include sterile aqueous solutions, dispersions, and sterile powders for preparing sterile solutions. Media and agents for preparing pharmaceutically active substances are well known in the art. If any conventional media or agents are incompatible with the active compound, they are not considered for use in the pharmaceutical composition of the present invention. In addition, supplementary active compounds may also be incorporated into the composition.

[0064] Preparations or compositions suitable for administering the pharmaceutical compositions and compounds of the present invention to subjects are within the scope of the present invention. Any suitable route of administration may be used, such as parenteral, intravenous, subcutaneous, intramuscular, intracranial, intraorbital, ocular, intraventricular, intracapsular, intraspinal, intrathecal, intracisternal, intraperitoneal, intranasal, aerosol, topical or oral administration.

[0065] The present invention also relates in part to a method of providing adjuvant therapy or neoadjuvant therapy in combination with another cancer therapy for treating cancer in a subject in need thereof; the method comprises administering to a subject in need thereof a therapeutically effective amount of a compound, composition, or formulation thereof of the present invention to treat cancer in the subject.

[0066] Typically, an effective amount of the compound or composition of the present invention will be administered to a subject. In this article, the term "subject" includes mammals, preferably human or animal subjects, but most preferably human subjects. For pharmaceutical compositions, an effective amount of the compound of the present invention can be provided alone, or in combination with other compounds, or can be connected to a suitable carrier and / or other molecules (e.g., lipids).

[0067] In certain embodiments, the pharmaceutical composition or compound of the present invention may be provided in the form of a kit, optionally together with a carrier, and accompanied by instructions for use.

[0068] The "effective amount" of the compound or pharmaceutical composition of the present invention includes a therapeutically effective amount. "Therapeutically effective amount" refers to an amount that can achieve the desired therapeutic effect (e.g., treatment of cancer) at a specific dose and within a specific time. The therapeutic results can be, for example, preventing cancer metastasis, preventing cancer recurrence, reducing cancer marker levels, reducing tumor size, inhibiting targeted metabolic pathways, delaying pathological progression associated with cancer, or other methods for determining the therapeutic effect. The therapeutically effective amount of a compound can vary according to a number of factors, such as the disease state, the age, sex, and weight of the individual, the ability of the compound to induce a desired response in the individual, previous treatment experience, the nature and severity of the cancer to be treated, the route of administration, and the form of the composition. The dosage regimen can be adjusted to provide the best therapeutic effect. A therapeutically effective amount also refers to the amount in which the therapeutic beneficial effects of a compound exceed any toxic or deleterious effects thereof.

[0069] The compounds or compositions of the present invention are intended to be used as adjuvant therapy or neoadjuvant therapy. As used herein, the term "adjuvant therapy" refers to an additional cancer treatment used in conjunction with or after a primary treatment to reduce the risk of cancer recurrence. Primary treatment may include chemotherapy, radiotherapy, hormone therapy, targeted therapy, or biological therapy. As used herein, the term "neoadjuvant therapy" refers to a treatment performed before primary treatment (usually surgery) as the first step of treatment to shrink a tumor or reduce cancer. Examples of neoadjuvant therapy include chemotherapy, radiotherapy, and hormone therapy. Therefore, the compounds and compositions of the present invention are intended to be used in combination with other cancer therapies, including chemotherapy, radiotherapy, hormone therapy, targeted therapy, or biological therapy.

[0070] The amount of 2-hydroxypropyl-β-cyclodextrin (HPβCD) in the composition may vary according to factors such as the stage of cancer, the age, sex and weight of the individual. The dosage regimen may be adjusted to provide the best therapeutic effect. For example, a single dose may be administered, or multiple doses may be administered over a period of time. In order to facilitate administration and ensure uniformity of dosage, it may be more advantageous to prepare the composition in the form of a unit dose. The dosage of the composition of the present invention may be determined by techniques known to those skilled in the art or by methods taught herein.

[0071] Dosage values ​​may be varied and adjusted over time according to the individual need and the judgment of the person administering or supervising the administration of the pharmaceutical compositions or compounds of the present invention. For ease of administration and to ensure uniformity of dosage, it may be advantageous to prepare the composition in unit dosage form.

[0072] The compounds and compositions of the present invention are intended to treat cancer, prevent cancer metastasis, or prevent cancer recurrence.

[0073] The term "prevention" when used in connection with a medical disease or condition is art-recognized and includes administering a composition that reduces the frequency of occurrence or delays the onset of symptoms of the condition in a subject compared to a subject not receiving the composition.

[0074] The term "therapeutic" treatment is well known to those skilled in the art and includes administering one or more pharmaceutical compositions or compounds of the present invention to a subject. If the composition is administered after an adverse condition has already occurred, the treatment is a therapeutic treatment (i.e., it is intended to alleviate, improve or stabilize an existing adverse condition or its side effects).

[0075] Toxicity and therapeutic efficacy of the compositions of the invention can be determined by standard pharmaceutical procedures in cell culture or using experimental animals, such as determining LD 50 and ED 50 The data obtained from cell culture and / or animal studies can be used to formulate a dosage range suitable for use in subjects. The dosage of any composition of the invention is preferably in the range of ED50 However, the concentration of the blood circulation is within the range of almost no toxicity or no toxicity. The dosage can vary within this range, depending on the dosage form used and the route of administration used. For the compounds and compositions of the present invention, the therapeutically effective dose can be estimated first by cell culture assays.

[0076] The following examples are used to illustrate the present invention but are not intended to limit the present invention.

[0077] Example 1

[0078] Three different mechanisms of action were used to induce epithelial-mesenchymal transition in breast cancer and colorectal adenocarcinoma cell lines. Breast cancer MCF-7 cells were treated with 150 micromolar (μM) cobalt chloride (CoCl2) for 48 hours. First, a working stock solution of 2500 μM CoCl2 was prepared by dilution with distilled water. The resulting solution was further diluted before application to cells cultured in cell culture medium. Cell culture medium composition: Dulbecco's modified Eagle's medium (DMEM) (Sigma Aldrich, UK), supplemented with 10% fetal bovine serum (FBS) (Celtic Molecular Diagnostics, South Africa) and 1% penicillin-streptomycin (SigmaAldrich, UK). NMuMG cells (normal mouse mammary epithelial cell line) were treated with 10 ng / mL transforming growth factor β1 (TGF-β) for 48 hours. The stock solution of TGF-β was prepared by resuspending 2 μg of recombinant TGF-β in sterile 4 mM HCl, containing 1 mg / mL bovine serum albumin (BSA), and diluted to 20 μg / mL in water. NMuMG cells were cultured in Dulbecco's modified Eagle's medium (DMEM) (Thermo Fisher Scientific, USA) supplemented with 10% FBS, 1% penicillin-streptomycin, 1% GlutaMAX and 10 μg / mL insulin. Colorectal cancer HT-29 cells were treated with 50 ng / mL interleukin-6 (IL-6) for 24 hours. A working stock solution of 1 μg / mL IL-6 was prepared by diluting with 0.1% BSA in phosphate buffered saline (PBS). The resulting solution was further diluted before application to cells cultured in cell culture medium. Cell culture medium composition: McCoy's 5a (modified) medium (Sigma Aldrich, UK) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin.

[0079] Immunofluorescence microscopy was used to detect protein expression of epithelial-mesenchymal transition molecular markers in the control group and cells treated with CoCl2, TGF-β, and IL-6. The epithelial cell junction protein, E-cadherin, was used as a molecular marker to indicate cells with an epithelial phenotype, while the intermediate filament protein, vimentin, was used as a molecular marker to indicate cells with a mesenchymal phenotype. E-cadherin expression was detected by Alexa Fluor 488 staining (green), and cell nuclei were stained with DAPI (blue) ( Figure 1 ). Vimentin expression was detected by Texas Red staining (red), and cell nuclei were stained with DAPI (blue) ( Figure 2 ). Image by Floid TM The images were acquired using a cell imaging system and analyzed using Image J software.

[0080] from Figure 1 and Figure 2 It can be clearly seen that after the induction of epithelial-mesenchymal transition by CoCl2, TGF-β, and IL-6, the expression of E-cadherin was significantly decreased, while the expression of vimentin was significantly increased. Therefore, under all treatment conditions, the expression of epithelial marker E-cadherin was significantly decreased, while the expression of mesenchymal marker vimentin was increased.

[0081] Example 2

[0082] Findings from laboratory studies have shown that after successful induction of epithelial-mesenchymal transition in breast cancer (MCF-7 and NMuMg) and colorectal cancer (HT-29) cells, cholesterol depletion using the compound HPβCD resulted in increased expression of the epithelial marker E-cadherin and decreased expression of the mesenchymal marker vimentin, a change confirmed by immunofluorescence staining. Methyl-β-cyclodextrin (MβCD) was used as a positive control because its role as a cholesterol depletion agent has been extensively studied and reported. After a 2-hour treatment, cells were fixed (4% formaldehyde, 10 minutes); permeabilized (0.1% Triton-X-100, 10 minutes); and a blocking step (1% bovine serum albumin in 0.1% Tween-20 solution, 1 hour) was completed. Cells were incubated with primary antibodies overnight at 4°C (dilution 1:250), then washed with phosphate-buffered saline (PBS) and incubated with secondary antibodies for 45 minutes at room temperature (dilution 1:1000). After completing the nuclear counterstaining, the coverslip containing the adherent cells was mounted and Floid TMVisualization analysis was performed using a cell imaging system (Thermo Fisher Scientific, USA). Quantitative analysis of immunofluorescence intensity was performed using Image J software (National Institutes of Health, USA). Analysis of target cells and background fluorescence was based on the following parameters: area, integrated density, and mean gray value. Corrected total cellular fluorescence (CTCF) was calculated according to the following formula: CTCF = integrated density – (area of ​​selected cells × mean of background fluorescence readings).

[0083] The protein expression levels of E-cadherin were investigated in MCF-7, NMuMg, and HT-29 cells treated with different cholesterol-targeting drugs before and after epithelial-mesenchymal transition induction. E-cadherin expression was determined by Alexa Fluor 488 staining (green), and nuclei were stained with DAPI (blue).

[0084] The protein expression levels of vimentin in MCF-7, NMuMg, and HT-29 cells treated with different cholesterol-targeted drugs before and after epithelial-mesenchymal transition induction were also investigated. Vimentin expression was determined by Texas Red staining (red), and cell nuclei were stained with DAPI (blue). Images courtesy of Floid TM The images were acquired by a cell imaging system and analyzed using Image J software. In addition, the expression levels of E-cadherin, vimentin, and N-cadherin were further analyzed by whole protein lysate in the NMuMg cell line to verify these experimental results.

[0085] Based on these findings, it can be seen that cholesterol-targeted drug treatment leads to increased expression of E-cadherin, an effect that is particularly pronounced in cells undergoing epithelial-mesenchymal transition ( Figure 3 ). In addition, it was observed that cholesterol-targeted drug treatment led to a decrease in vimentin expression, an effect that was particularly pronounced in cells undergoing epithelial-mesenchymal transition ( Figure 4 ).

[0086] These results are highly consistent with the Western blot data obtained in the NMuMg cell line. 6 Each cell was lysed with 100 μL of lysis buffer. Protein lysate was quantified using the bicinchoninic acid (BCA) protein quantification method (Sigma Aldrich, UK) and the bovine serum albumin (BSA) (Inqaba Biotec TM The protein lysate with a final concentration of 10 mg / mL was loaded onto a 10% SDS-PAGE gel and eluted using a semi-dry transfer system. Turbo TMThe protein was transferred to a PVDF membrane (pore size 0.22 μm) using a transfer system (Bio-Rad, USA). The PVDF membrane was then blocked with 3% BSA for 1 hour. The primary antibody was added and incubated overnight. The secondary antibody was added the next day and incubated for 1 hour. TM Western ECL HRP substrate (Bio-Rad, USA) was used to visualize protein bands on a ChemiDoc imaging system (Bio-Rad, USA).

[0087] The expression levels of epithelial marker E-cadherin and mesenchymal markers vimentin and N-cadherin were analyzed, and the results showed that the expression of these markers was reduced after treatment with cholesterol-targeted drugs ( Figure 5 ). In addition, in the NMuMg cell line, cholesterol-targeted drug treatment significantly reduced the expression of CTBP1 (master regulator) and SMAD4 (effector molecule), both of which play key roles in the TGF-β signaling pathway. This result supports the observation that the expression levels of epithelial-mesenchymal transition-related transcription factors TWIST1 and ZEB1 were reduced after cholesterol-targeted drug treatment. Therefore, it can be speculated that targeting cholesterol in cells after epithelial-mesenchymal transition helps to promote the restoration of the epithelial phenotype.

[0088] Example 3

[0089] The number of invasive cells in MCF-7, NMuMg, and HT-29 cells treated with different cholesterol-targeting drugs before and after induction of epithelial-mesenchymal transition was investigated, and the nuclei were stained with DAPI (blue) for visualization. To achieve this, after treatment, the cells were trypsinized and resuspended in serum-free medium and then seeded in Geltrex-coated Transwell chambers (pore size 0.8 μm). The Transwell chambers were placed in 24-well plates and incubated at 37°C for 24 hours. After the incubation period, non-invasive cells were removed from the top of the chamber with a cotton swab, and the invasive cells were fixed (4% formaldehyde, 10 minutes) and stained with DAPI. Image courtesy of Floid TM The images were acquired using a cell imaging system and analyzed using Image J software.

[0090] Depend on Figure 6The results showed that epithelial-mesenchymal transition led to a significant increase in the invasive potential of MCF-7, NMuMg, and HT-29 cells. Treatment with cholesterol-targeted drugs significantly reduced the invasive potential of cells after epithelial-mesenchymal transition and may effectively reduce the metastatic potential of cancer cells. This effect may be attributed to the disruption of lipid raft integrity, which in turn promotes the shedding of key cell surface receptors that cancer cells rely on to confer an invasive disease phenotype. In addition, multiple epithelial-mesenchymal transition-related signaling pathways were also inhibited. This effect promoted the increased expression of key epithelial markers while reducing the expression of mesenchymal markers. Therefore, the application of cholesterol-targeted drugs can effectively inhibit epithelial-mesenchymal transition and cancer cell metastasis and prevent them from acquiring an invasive disease phenotype.

[0091] To elucidate the molecular mechanism of cholesterol-mediated epithelial-mesenchymal transition induction, RT-qPCR analysis of major cholesterol-related genes was performed in the NMuMg cell line. TM RNA MiniPrep Kit (ZymoResearch, Inqaba Biotec TM , South Africa) to extract RNA. RNA extraction was performed according to the manufacturer's instructions. 1 μg of total RNA was used for cDNA synthesis using the RevertAid First Strand cDNA Synthesis Kit (Thermo Fisher Scientific, USA) according to the manufacturer's instructions. SensiFAST TM No-ROX Kit, in CFX96 Touch TM A three-step qPCR experiment was performed on a real-time fluorescence quantitative PCR detection system (Bio-Rad).

[0092] Increased expression of LXR, a master regulator of cholesterol efflux, was documented, supporting the observation of decreased expression of the master cholesterol efflux genes ABCA1 and ABCG1. In addition, increased expression of the master regulator SREBF2 confirmed the increased expression of HMGCR, LDLR, and PCSK9 in cells after epithelial-mesenchymal transition. The above observed effects were reversed after treatment with cholesterol-targeted drugs, with MβCD having a more significant effect on cholesterol biosynthesis and uptake. In contrast, HPβCD and simvastatin had a significant effect on cholesterol efflux.

[0093] To assess the potential for multidrug resistance in cells before and after epithelial-mesenchymal transition, we used MDR detection kit (Thermo Fisher Scientific, USA). The test is based on the administration of the appropriate multidrug resistance protein 1 (MDR1) substrate calcein acetoxymethyl ester (calcein-AM), and the drug efflux capacity is evaluated by detecting the fluorescence level of intracellular calcein. In a 96-well black-walled plate, 10,000 cells were seeded per well. After 2 hours of treatment, the cells were incubated with 0.25 μM calcein-AM for 30 minutes at 37°C. Subsequently, PBS was washed twice, and PBS was added to each well to determine the fluorescence intensity. The Victor Nivo multi-mode microplate reader (Perkin Elmer, USA) was used, with the maximum excitation wavelength set to 494 nm and the maximum emission wavelength set to 517 nm.

[0094] In MCF-7 and HT-29 cells, intracellular retention of fluorescent calcein after treatment with cholesterol depleting agents was significantly increased following epithelial-mesenchymal transition, thus reinforcing the use of cholesterol-targeted drugs as a suitable means of inhibiting the potential for multidrug resistance. This effect may be attributed to changes in lipid raft membrane structure, resulting in the translocation of MDR1 to the detergent-soluble fraction, which in turn affects the basal ATPase activity. This in turn affects the efflux capacity of the MDR1 transporter. Preliminary data from HT-29 cells showed that cholesterol depleting agents increased the intracellular retention of fluorescent calcein in cells treated with the FOLFOX chemotherapy regimen. Fluorescence intensity (RFU) was quantified using a Victor Nivo multimode microplate reader (Perkin Elmer, USA) with the maximum excitation wavelength set at 494 nm and the maximum emission wavelength set at 517 nm.

[0095] In summary, these results show that cholesterol depletion treatment significantly reduces the invasion and drug resistance potential of cells. The above experimental results show that after cholesterol depletion using HPβCD, the epithelial-mesenchymal transition phenotype is reduced. Through this reversal of epithelial-mesenchymal transition, the inventors showed a reduction in invasion potential and drug resistance. In addition, it was shown that HPβCD has lower toxicity compared to simvastatin, a statin that blocks cholesterol synthesis.

[0096] Example 4

[0097] To investigate the in vivo application of HPβCD as an anticancer drug for colorectal cancer (CRC), NOD / SCID mice were used for experiments. 24 NOD / SCID mice (5-8 weeks old) were injected with 4×10 6 HT-29 cells (suspended in 10% McCoy's medium and 1× PBS) were added until tumors formed. When the tumor size reached 150–200 mm 3Treatment was started at 1-2 weeks (a total of 12 treatments per group, but only 6 treatments were performed in the OXAL treatment group due to significant toxicity). The treatment groups included 4 mice each and were divided into the following groups: control group (injected with 1× PBS), HPβCD treatment group, oxaliplatin (OXAL) treatment group, 5-fluorouracil (5FU) treatment group, HPβCD combined with OXAL treatment group, and HPβCD combined with 5FU treatment group. Mice were intraperitoneally injected with 3000 mg / kg HPβCD, 10 mg / kg OXAL, and 30 mg / kg 5FU, 3 times a week for 4 weeks. For the combination treatment, HPβCD was administered simultaneously with 5-FU or OXAL chemotherapy. The body weight of mice was measured three times a week, and the average of the weekly body weight was calculated. When the mice were euthanized, serum was collected from whole blood samples and further used to measure cholesterol levels, as well as aspartate aminotransferase (AST) and alanine aminotransferase (ALT) as indicators of hepatotoxicity. Perfusion was then performed, and all organs, including tumors, were collected. All organs were snap frozen and stored at -80°C. Tumor samples were further stored in 10% neutral buffered formalin (4°C) and RNA Later (-80°C).

[0098] A significant reduction in tumor size and weight was observed. A reduction in tumor size was observed after HPβCD treatment, and further reduction in tumor size was observed after combined treatment ( Fig. 9 Compared with the OXAL alone treatment group, the tumor size of mice treated with HPβCD was reduced by an average of about 17%, while the tumors treated with HPβCD combined with OXAL were further reduced by 31%. In addition, HPβCD showed a total of about 74% reduction in tumor size and weight in both combined treatment groups (Table 1 and Fig.10 ). Therefore, the combination of HPβCD and OXAL or HPβCD and 5FU showed a further reduction in tumor size compared to these chemotherapeutic drugs alone. Therefore, it can be inferred that HPβCD enhances the efficacy of these drugs. Interestingly, HPβCD reduced the toxicity of OXAL, as the OXAL alone treatment group became toxic after only 6 treatments (2 deaths and 2 required euthanasia). The OXAL and HPβCD combination treatment group was able to complete the treatment regimen (12 treatments at the same dose) and the mice were in good health. After death, autopsy showed that one mouse in the OXAL alone treatment group had metastases almost all over the body, especially in the liver.

[0099] The weight of mice was measured three times a week, and the average of the weekly weight was calculated. The weight was monitored throughout the experiment as one of the indicators of the health status of the mice. Fig.11 shown.

[0100] Table 1: Results in the table show the tumor size and weight of mice in each treatment group, demonstrating the increase or decrease in tumors compared to the untreated group.

[0101]

[0102] HPβCD-treated mice had slightly higher serum cholesterol levels (average 65.5 mg / dL) compared with untreated mice (average 51 mg / dL). Fig.12 ). This temporary increase in cholesterol levels is expected because HPβCD has cholesterol sequestration properties that can excrete excess cholesterol into the blood. Interestingly, mice treated with OXAL and 5FU had significantly increased serum cholesterol levels. However, combined treatment with HPβCD showed a decrease in serum cholesterol levels in mice, which may lead to a decrease in cholesterol uptake in these cancer cells, thereby improving the efficacy of OXAL and 5FU in inhibiting tumor growth. Another possible explanation is that cyclodextrin, as a known excipient, increases the solubility of the drug. Conversely, OXAL and 5FU may increase the activity of transporters, thereby enhancing drug efflux and may also enhance cholesterol efflux. However, combined treatment with HPβCD reduced serum cholesterol levels in mice. Cells may use more cholesterol, or the liver may secrete cholesterol in the form of bile acids. The graph shows the changes in cholesterol levels in the blood of a group of mice. For the OXAL treatment group, the inventors were only able to draw blood from one mouse because the rest of the mice died due to OXAL toxicity.

[0103] Blood was collected after euthanasia of mice, and serum AST and ALT levels were analyzed by IDEXX Catalyst DX chemistry analyzer. ALT is a more specific indicator of hepatotoxicity because its concentration in the liver is higher than in other tissues. Fig.13 The hepatotoxicity markers ALT and AST levels and their ratios are shown. ALT measurements were similar between the untreated group and all treated groups ( Fig.13 , upper figure). No hepatotoxicity was observed in the HPβCD treated group (mean 1.67 U / L) compared to the untreated group (mean 1.59 U / L). An AST:ALT ratio greater than 2 indicates hepatotoxicity, which was only observed in the HPβCD+OXAL treated group ( Fig.13, lower figure). The AST:ALT ratio was not available for the OXAL monotherapy group. However, it is speculated that HPβCD reduced the toxicity of OXAL monotherapy. This was because after 6 doses of OXAL monotherapy, 50% of the mice in this treatment group died unexpectedly. The remaining mice in the OXAL treatment group were euthanized according to ethical regulations. Due to the unexpected termination of this group, no appropriate blood samples were obtained. Mice treated with the combination of OXAL and HPβCD were able to tolerate 12 doses of OXAL without significant adverse health effects.

[0104] In addition, HPβCD showed reduced toxicity associated with 5FU. No metastasis was observed in the HPβCD co-treatment group. Most mice also did not develop liver toxicity. The temporary increase in cholesterol levels in the HPβCD-treated group indicated that the treatment was working because HPβCD was extracting cholesterol from the cells. Therefore, HPβCD could be a good combination with existing chemotherapy drugs to reduce their toxicity, improve efficacy, prevent metastasis, and reduce tumor resistance.

Claims

1. 2-Hydroxypropyl-β-cyclodextrin (HPβCD) for use in a method of treating cancer, the method comprising administering the HPβCD to a subject in need thereof in combination with another cancer therapy.

2. HPβCD for use according to claim 1, wherein The HPβCD is administered as a neoadjuvant prior to another cancer therapy.

3. HPβCD for use according to claim 2, wherein Administration of the HPβCD increases the sensitivity of cancer cells to the cancer therapy by reversing the epithelial-mesenchymal transition phenotype of cancer cells.

4. HPβCD for use according to claim 2 or 3, wherein Administration of the HPβCD improves the subject's response to the cancer therapy.

5. HPβCD for use according to claim 1, wherein The HPβCD is administered as an adjunct in conjunction with or after another cancer therapy.

6. HPβCD for use according to claim 5, wherein Administration of the HPβCD sensitizes the non-proliferating cancer cells to the cancer therapy.

7. HPβCD for use according to claim 5 or 6, wherein Administration of the HPβCD reduces the toxicity of the cancer therapy and / or reduces one or more side effects of the cancer therapy.

8. HPβCD for use according to any one of claims 5 to 7, wherein Administration of the HPβCD reduces the risk or incidence of cancer recurrence.

9. HPβCD for use according to any one of claims 1 to 8, wherein The cancer therapy is selected from the group consisting of chemotherapy, radiotherapy, hormone therapy, targeted therapy or biological therapy.

10. A method of treating cancer comprising administering 2-hydroxypropyl-β-cyclodextrin (HPβCD) in combination with another cancer therapy to a subject in need thereof.

11. The method of claim 10, wherein: The HPβCD is administered as a neoadjuvant prior to another cancer therapy.

12. The method of claim 11, wherein: Administration of the HPβCD increases the sensitivity of cancer cells to the cancer therapy by reversing the epithelial-mesenchymal transition phenotype of cancer cells.

13. The method according to claim 11 or 12, wherein: Administration of the HPβCD improves the subject's response to the cancer therapy.

14. The method of claim 10, wherein: The HPβCD is administered as an adjunct in conjunction with or after another cancer therapy.

15. The method of claim 14, wherein: Administration of the HPβCD sensitizes the non-proliferating cancer cells to the cancer therapy.

16. The method according to claim 14 or 15, wherein: Administration of the HPβCD reduces the toxicity of the cancer therapy and / or reduces one or more side effects of the cancer therapy.

17. The method according to any one of claims 14 to 16, wherein: Administration of the HPβCD reduces the risk or incidence of cancer recurrence.

18. The method according to any one of claims 10 to 17, wherein: The cancer therapy is selected from the group consisting of chemotherapy, radiotherapy, hormone therapy, targeted therapy or biological therapy.

19. Use of 2-hydroxypropyl-β-cyclodextrin (HPβCD) in the preparation of a medicament for use in a method of treating cancer comprising administering the medicament to a subject in need thereof in combination with another cancer therapy.

20. The use according to claim 19, wherein The medicament comprising HPβCD is administered as a neoadjuvant prior to another cancer therapy.

21. The use according to claim 20, wherein Administration of the medicament comprising HPβCD increases the sensitivity of cancer cells to the cancer therapy by reversing the epithelial-mesenchymal transition phenotype of cancer cells.

22. The use according to claim 20 or 21, wherein Administration of the medicament comprising HPβCD improves the subject's response to the cancer therapy.

23. The use according to claim 19, wherein The medicament comprising HPβCD is administered as an adjunct in combination with another cancer therapy or after another cancer therapy.

24. The use according to claim 23, wherein Administration of the medicament comprising HPβCD sensitizes the non-proliferating cancer cells to the cancer therapy.

25. The use according to claim 23 or 24, wherein Administration of the HPβCD reduces the toxicity of the cancer therapy and / or reduces one or more side effects of the cancer therapy.

26. The use according to any one of claims 23 to 25, wherein Administration of the medicament comprising HPβCD reduces the risk or incidence of cancer recurrence.

27. The use according to any one of claims 19 to 26, wherein The cancer therapy is selected from the group consisting of chemotherapy, radiotherapy, hormone therapy, targeted therapy or biological therapy.

28. A pharmaceutical composition comprising 2-hydroxypropyl-β-cyclodextrin (HPβCD) and a pharmaceutically acceptable carrier, and optionally a chemotherapeutic drug.

29. The pharmaceutical composition of claim 28, further comprising a pharmaceutically acceptable excipient, diluent, carrier or other suitable additives.

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