EA046190B1 - NEW COMBINED SOLUTION FOR THE TREATMENT OF CANCER RESISTANT TO CHEMOTHERAPY - Google Patents

Description

This invention claims priority to PRC Provisional Patent Application CN 201811195239.3, filed on October 12, 2018, the contents of which are incorporated herein by reference in their entirety.

Technical field

The present invention relates to a combination of drugs for the treatment of chemotherapy-resistant cancer.

State of the art

Today, cancer remains the leading cause of death worldwide. Although people have made significant progress in treating some forms of cancer through the use of new methods of surgery, radiation therapy and chemotherapy, cancer remains largely incurable. Even if an effective treatment is available for a particular form of cancer, the side effects from such treatment can be quite severe, leading to an undesirable decrease in the patient's quality of life, and the cancer always develops resistance to the effects of chemotherapy.

Most conventional chemotherapy drugs have significant toxicity and only limited efficacy, especially in patients with advanced solid tumors. And while traditional chemotherapy drugs kill cancer cells, they also damage normal, healthy human cells. The therapeutic index (i.e., a measure of a given treatment's ability to distinguish cancer cells from normal cells) for such chemotherapy drugs can be quite low. Very often, the dose of chemotherapy that effectively kills cancer cells also kills normal cells, especially those that undergo frequent cell division. When such therapy affects normal cells, side effects often occur, manifested in the form of hair loss, suppression of hematopoietic function and nausea. Depending on the patient's overall health, these adverse events may cause the cancer patient to experience at least very unpleasant side effects or even cause him or her to stop taking chemotherapy drugs altogether. Even if a patient with cancer initially responds well to chemotherapy with visible tumor shrinkage, it is very common for the cancer to recur quickly. These recurrent cancers often become highly resistant or resistant to chemotherapy.

The so-called cancer stem cells (CSC (Cancer Stem Cells), which are also often called, for example, tumor-initiating cells, tumor stem-like cells, cancer stem-like cells, high-grade cells, are responsible for the rapid relapse of the tumor and its development of resistance to further action of traditional chemotherapy. tumorigenic potential or super-malignant cells). There is growing evidence to suggest that CSC cells are present in virtually all tumor types as distinct populations and that they give rise to specialized cells that form the bulk of the tumor mass and phenotypically characterize the disease. CSC cells have been shown to be fundamentally responsible for carcinogenesis, metastasis formation, recurrence and recurrence of cancer (see, for example, Fig. 1). CSC cells are naturally resistant to conventional chemotherapy, which means that they remain undamaged by conventional chemotherapy that kills the bulk of tumor cells (see, for example, Fig. 2). Based on this, the very fact of recognizing the existence of CSC cells entails a number of consequences regarding the further development of cancer treatment methods and the improvement of therapeutic methods. This includes, for example, disease identification, selection of drug targets, prevention of cancer metastasis and recurrence, treatment of cancers that are resistant to chemotherapy and/or radiation therapy, treatment of cancers that are naturally resistant to chemotherapy or radiation therapy, and development new strategies to combat cancer.

The STAT3 signaling protein is a potent transcriptional regulator targeting a large number of genes involved in the cell cycle, cell survival, tumorigenesis, tumor progression and metastasis, including but not limited to BCL-XL, c-MYC, CYCLIN D1, IDO1, PDL1, VEGF, MMP-2 and SURVIVIN (see, for example, Fig. 3). Collective expression of these STAT3-responsive genes maintains the stemness of cancer stem cells (CSCs), essential for the survival and proliferation of such stem cells. Therefore, STAT3 may play a key role in the ability of CSC cells to survive and self-renew in a wide range of cancers. Accordingly, STAT3 is emerging as a promising target to address the challenges of suppressing cancer stem cell survival and preventing the formation of metastases. Drugs that specifically target the STAT3 signaling protein and are active against CSC cells hold great promise for cancer patients (Boman, B.M., et al. J. Clin. Oncol., 2008. 26(17): p. 2795-99 ).

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According to the PCT patent application WO 2009/036059, a compound of formula (I)

o is an inhibitor of CSC cell growth and survival.

The compound of formula (I) inhibits the activity of STAT3 to control the transcription algorithm with a cellular half-maximal inhibitory concentration IC ₅₀ of the order of ~0.25 μM. The compound of formula (I) can be synthesized according to patent application according to PCT WO 2009/036059, see example 13. In some embodiments, the compound of formula (I) is used in a method of treating cancer. According to the patent application according to PCT WO 2014/169078, example 6, the compound of formula (I) was selected for clinical trials on patients with advanced stages of cancer. According to patent application PCT WO 2009/036101, a regimen of combining two drugs, one of which is a compound of formula (I) and the other a conventional antitumor drug, produces a potential synergistic antitumor effect. Prospects for the development of new antitumor strategies based on the suppression of the CSC cell-controlled transcription algorithm to prevent the formation of metastases and relapses of cancer, to treat cancers that have become resistant to chemotherapy and/or radiation therapy, and to treat cancers that are inherently resistant to chemotherapy and/or radiation therapy are associated with an urgent need for further development of new, alternative and more effective combinations of various therapeutic regimens based on the compound of formula (I) for the treatment of patients with specific types of cancer.

Pancreatic cancer is a fairly common type of cancer, characterized by rapid progression of the disease and poor prospects for the patient, since it is hidden and difficult to identify in the early stages of the disease, the tumor is difficult to remove surgically and easily recurs after surgery, and chemotherapy is ineffective against it. . Currently, the only possible treatment for such cancer is surgery. However, more than 80% of patients with metastatic pancreatic cancer already have locally advanced or metastatic cancer at the time of diagnosis. Of the small number of patients who do receive surgical treatment in this setting, the majority will progress to more advanced stages of pancreatic cancer, with five-year survival rates of less than 5% (Hidalgo, 2010). Currently, standard first-line chemotherapy regimens for patients with unresectable metastatic pancreatic cancer include FOLFIRINOX (5-fluorouracil (5-FU), leucovorin/LV), irinotecan, and oxaliplatin for combination chemotherapy. For untreated patients, the median overall survival (mOS) is 11.1 months (Conroy, 2011), and a combination chemotherapy regimen using gemcitabine in combination with nabpaclitaxel, which was recently used in a phase II/III clinical trial MRAST (Von Hoff, 2013), the mOS in the combined group was 8.7 months, while this combined group had an unprecedented three-year survival rate of about 4% (Goldstein, 2014). For patients whose disease continues to develop after receiving first-line treatment, the options currently available for further treatment are very limited. For example, for those who have not used gemcitabine in first-line treatment, standard doses of gemcitabine can be used in second-line treatment. For those who have already used gemcitabine in first-line treatment, 5-FU/LV is usually the standard choice for second-line treatment. The mOS in patients with pancreatic cancer after failed first-line treatment is approximately 4-5 months. Recent studies of a therapeutic regimen using Onivyde in combination with 5-FU/LV show that for patients with metastatic pancreatic cancer that continued to progress after first-line gemcitabine-based treatment, the mOS value of the combination regimen increased to 6.1 months (Chen, 2015). Researchers continue to work hard, and there is an urgent need to develop new treatment regimens that would improve the survival picture for patients with metastatic pancreatic cancer, especially those who have failed first-line treatment.

Brief description of the invention

The present invention discloses uses of a novel combination of drugs for the treatment of chemotherapy-resistant cancer, wherein said combination of drugs comprises a therapeutically effective amount of a compound of formula (I)

- 2 046190 o

o tD or its pharmaceutically acceptable salts or solvates, a low dose of gemcitabine, or its pharmaceutically acceptable salts or solvates, and a therapeutically effective amount of paclitaxel, or its pharmaceutically acceptable salts or solvates.

In developing the present invention, it was unexpectedly discovered that in a therapeutic regimen using a compound of formula (I) or pharmaceutically acceptable salts or solvates thereof in combination with low doses of gemcitabine or pharmaceutically acceptable salts or solvates thereof and paclitaxel or pharmaceutically acceptable salts or solvates thereof, the compound of formula (I) provides antitumor activity and sustained response in patients with metastatic pancreatic cancer whose previous treatments have failed.

Certain embodiments disclosed herein provide methods of treating cancer comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound of formula (I) or pharmaceutically acceptable salts or solvates thereof, a low dose of gemcitabine or pharmaceutically acceptable salts or solvates thereof, and therapeutically an effective amount of paclitaxel or pharmaceutically acceptable salts or solvates thereof.

The compound of formula (I), low dose gemcitabine and paclitaxel can be administered to a patient simultaneously, in parallel, separately and/or sequentially. Thus, in some embodiments, a compound of Formula (I) and a low dose of gemcitabine are administered to a patient simultaneously, in parallel, separately, and/or sequentially. In some other embodiments, a compound of formula (I) and paclitaxel are administered to a patient simultaneously, in parallel, separately, and/or sequentially.

The compound of formula (I) can be administered as a single dose or as a divided total dose daily. A therapeutically effective amount of a compound of formula (I) is administered daily at a dose of about 80-960 mg and about 80-480 mg. A compound of formula (I) is administered twice daily at a dose of about 80 mg, about 160 mg, or about 240 mg. Paclitaxel can be administered weekly. In this embodiment, paclitaxel m can be administered in an amount of 10-100 mg/m ² every week. In this case, a therapeutically effective amount of paclitaxel is administered weekly in doses of about 80 mg/m ² , about 60 mg/m ² and about 40 mg/m ² by intravenous infusion. A low dose of gemcitabine can be administered weekly. In this embodiment, low dose gemcitabine can be administered in an amount of 100-800 mg/m ² every week, and this dose of gemcitabine is less than 50% of the recommended effective dose. In this case, low dose gemcitabine is administered weekly in amounts of about 600 mg/ ^m2 , about 550 mg/ ^m2 , about 500 mg/ ^m2 , about 450 mg/ ^m2 , about 400 mg/ ^m2 , about 300 mg/ ^m2 , about 200 mg/m ² or about 100 mg/m ² by intravenous infusion.

In some embodiments, the cancers are advanced, metastatic, non-surgical, refractory, or recurrent cancers. In some embodiments, the cancer is at least one pancreatic cancer that has continued to progress after a previous course of treatment. In some embodiments, the pancreatic cancer is pancreatic adenocarcinoma. In some embodiments, the pancreatic cancer is pancreatic ductal adenocarcinoma. In some embodiments, the pancreatic cancer is at least one pancreatic cancer that has continued to progress after a previous course of treatment.

In some embodiments, disclosed herein are the use of combinations of drugs in methods of increasing the responsiveness of a subject to at least one therapeutic regimen, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of formula (I), a selected compound of formula (I)

O (I) or its pharmaceutically acceptable salts and solvates.

In some embodiments, disclosed herein are methods of increasing the responsiveness of a subject to at least one therapeutic regimen, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of formula (I).

In some embodiments, disclosed herein are methods of increasing the responsiveness of a subject to at least one pre-therapy regimen, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound

- 3 046190 formula (I), selected from compounds of formula (I)

or pharmaceutically acceptable salts and solvates thereof.

In some embodiments, disclosed herein are the uses of drug combinations in methods of increasing the responsiveness of a subject to at least one pre-therapy regimen, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of formula (I).

In some embodiments, disclosed herein are the uses of combinations of drugs in methods of increasing the responsiveness of a subject to at least one pre-therapy regimen, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of formula (I) selected from compounds of formula (I )

or pharmaceutically acceptable salts and solvates thereof.

In some embodiments, the at least one pre-therapy regimen is selected from chemotherapy regimens. In some embodiments, the at least one pre-therapy regimen is selected from gemcitabine regimens. In some embodiments, the at least one pre-therapy regimen is selected from gemcitabine-based monotherapy or combination therapy. In some embodiments, the at least one pre-therapy regimen is selected from paclitaxel regimens. In some embodiments, the at least one pre-therapy regimen is selected from FOLFIRINOX, mFOLFIRINOX, or Gem-Abraxane (gemcitabine and albumin-stabilized paclitaxel). In some embodiments, the at least one pre-therapy regimen is selected from conventional cancer treatment regimens, such as surgery, radiation therapy, targeted therapy, or immunotherapy.

In some embodiments, disclosed herein are methods of increasing the responsiveness of a subject to a chemotherapy regimen, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of formula (I) selected from compounds of formula (I)

or pharmaceutically acceptable salts and solvates thereof, a low dose of gemcitabine and a therapeutically effective amount of paclitaxel.

In some embodiments, the subject is a patient suffering from pancreatic cancer with failed prior treatment. In some embodiments, the subject is a patient suffering from metastatic pancreatic cancer with failed prior treatment.

In some embodiments, disclosed herein are methods of increasing the responsiveness of a subject to a chemotherapy regimen, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of formula (I).

In some embodiments, disclosed herein are methods of inhibiting, shortening, and/or minimizing the survival and/or self-renewal of cancer stem cells and heterogeneous cancer cells, comprising administering to a subject in need of treatment a therapeutically effective amount of a compound of formula (I), low dose of gemcitabine and a therapeutically effective amount of paclitaxel.

In some embodiments, disclosed herein is a kit comprising at least one compound selected from compounds having formula (I), prodrugs, derivatives, pharmaceutically acceptable salts of any of the foregoing compounds, and solvates of any of the foregoing compounds. In some embodiments, disclosed herein is a kit comprising at least one gemcitabine selected from gemcitabine, prodrugs, derivatives, pharmaceutically acceptable salts, and solvates thereof of any of the foregoing. In some embodiments of the invention, a set is disclosed herein, including

- 4 046190 containing at least one paclitaxel selected from paclitaxel, its prodrugs, pharmaceutically acceptable derivatives, and solvates. In some embodiments, disclosed herein is a kit comprising at least one compound selected from compounds having Formula (I), prodrugs, derivatives, pharmaceutically acceptable salts of any of the foregoing compounds, and solvates of any of the foregoing compounds, at least one gemcitabine selected from gemcitabine, prodrugs, derivatives, pharmaceutically acceptable salts of any of the above compounds, and solvates of any of the above compounds, and at least one paclitaxel selected from paclitaxel, prodrugs, derivatives, pharmaceutically acceptable salts of any of the above compounds, and solvates of any of the above compounds.

Aspects and embodiments of the present invention are set forth in more detail and will be clearly expressed in the following detailed description of the invention. It should be understood that both the foregoing general description and the following detailed description are exemplary and explanatory in nature only and are not intended to limit the claims.

Brief description of drawings

Fig. 1 shows the formation of heterogeneous cancer cells from cancer stem cells.

Fig. 2 shows the effect of cancer stem cell-specific therapy and traditional anticancer therapy.

Fig. 3 shows the STAT3 transcriptional activator signaling pathway in cancer.

Fig. 4 shows typical effects of 2-acetylnaphtho[2,3-b]furan-4,9-dione, gemcitabine (Gemzar) and combination treatment of 2-acetylnaphtho[2,3-b]furan-4,9-dione and gemcitabine in a mouse xenograft tumor model according to specific embodiments of the present invention.

Fig. 5 shows typical effects of 2-acetylnaphtho[2,3-b]furan-4,9-dione, paclitaxel, and combination treatment of 2-acetylnaphtho[2,3-b]furan-4,9-dione and paclitaxel on cancer stemness markers p -Stat3 and β-catenin levels in pancreatic cancer stem cells (Panc-1) in vitro.

Detailed Description of the Invention

The following are definitions used in this specification. The definition first given to a group or term in this specification applies to that same group or individual term throughout the remainder of this specification, either individually or as part of other groups or terms, unless otherwise specified.

When the term is roughly used in connection with a range of numerical values, it indicates that the range may also be extended above the upper and below the lower bound of its numerical values. In general, the term approximately in this description is used to show that the numerical value in question can be changed up or down by deviating from its specified value by 20, 10, 5 or 1%. In some embodiments of the invention, the term is roughly used to indicate that the numerical value in question can be changed by deviating up or down from its specified value by 10%. In some embodiments of the invention, the term is roughly used to indicate that the numerical value in question can be changed by deviating more or less from its specified value by 5%. In some embodiments of the invention, the term is roughly used to indicate that the numerical value in question can be changed by deviating more or less from its specified value by 1%.

When the term less than is used in connection with a range of numerical values, it is intended to indicate the possibility of covering any arbitrary numerical value or subrange that is less than and does not overlap with the specified range. For example, less than 5 mg means that values such as 1, 2, 3, and 4 mg can be included. In some embodiments, the expression administer less than 600 mg/ ^m2 also means the ability to administer 599 mg/ ^m2 , administer 598 mg/m2 ^, etc.

The phrase and/or when used in this specification, in these guidelines and in the claims of this invention should be understood as either or both of the two simultaneously with respect to the elements in question associated with this phrase, i.e. elements that in some cases may be present together, and in other cases present separately. Thus, as a non-exhaustive example, consider the expression A and/or B. If used in conjunction with a non-limiting expression of the inclusive type, it may refer in one embodiment only to A (may also optionally include other elements, but different from B), in another embodiment only to B (can also optionally include other elements, but different from A), and in a third embodiment - to A and B simultaneously (can also optionally include other elements), etc. When a range of values is specified in this specification, this means that each value and subrange included within the given range is covered. For example, the expression 1-5 mg means to cover values and sub-ranges such as 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 1-2 mg, 1-3 mg, 1-4 mg, 1-5 mg, 2-3 mg, 2-4 mg, 2-5 mg, 3-4 mg, 3-5 mg and 4-5 mg.

The terms introduce, input or introduction are used in this description to their broadest extent.

- 5 046190 sense. These terms refer to any method of administering to a subject a chemical compound or pharmaceutical composition described herein and may include, for example, administering the agent systemically to a subject, i.e. in relation to the whole organism as a whole, locally or in situ. Thus, a compound claimed to be the subject matter of this invention that can be produced within a subject from a composition administered to him (whether or not including the compound) is also covered by these terms. When used in conjunction with the terms systemic or systemic, they generally refer to the in vivo systemic absorption or systemic accumulation of a given compound or composition in the bloodstream followed by distribution throughout the body.

The term subject generally refers to an organism to which a substance or pharmaceutical composition described herein can be administered. The subject may be a mammalian animal or a cell thereof, including a human or a human cell. The term also refers to an organism comprising a cell or being a donor or recipient of such a cell. In various embodiments of the present invention, the term subject refers to any animal (e.g., mammal), including, but not limited to, humans, mammals, and non-mammals such as non-human primates, mice, rabbits, sheep, dogs, cats, horses, cows , chickens, amphibians, reptiles, fish, nematodes and insects that will be required to act as recipients of the substance or pharmaceutical composition described herein. In some circumstances, the term subject is equivalent to the term patient, and both terms may be used equally herein to refer to a human subject.

The term therapeutically effective amount, as used herein, has its meaning generally accepted in the art. The term generally refers to the amount of a substance or composition that will produce a desired biological or medical response in a cell, tissue, system, animal or human. For example, if a clinical therapy in question is considered effective when it produces at least about 25% reduction in a measurable parameter that characterizes the disease or disorder, then the therapeutically effective amount of the drug used to treat the disease or disorder will be the amount required to obtain the effect of reducing this measured parameter by at least about 25%.

The term "therapeutically effective amount" in relation to the treatment of cancer means an amount of a drug capable of causing one or more of the following effects: (1) inhibiting, to varying degrees, the growth of cancer cells or malignant tumors, including slowing or completely stopping growth; (2) reduction in the number of cancer or tumor cells; (3) reduction in tumor size; (4) inhibition (ie, reduction, slowing, or complete cessation) of infiltration of cancer or tumor cells into peripheral organs; (5) suppression (ie, reduction, slowing or complete cessation) of the formation of metastases; (6) increasing the antitumor immune response, which may (but will not necessarily) lead to tumor regression or rejection; or (7) relief, to some degree, of one or more measurable symptoms associated with the presence of cancer or tumor. In some embodiments of the present invention, the term therapeutically effective amount refers to an amount administered systemically, locally, or in situ (eg, an amount of a substance that is produced in the body of a subject in situ). The therapeutically effective amount may vary depending on various factors, such as disease states, age, sex and weight of the individual, as well as the ability of one or more anticancer agents to produce the desired response in the individual. A therapeutically effective amount is also an amount at which the therapeutically beneficial effects outweigh any possible toxic or harmful effects.

Terms such as treat, cure, mitigate, alleviate, alleviate, as used herein, refer to (1) therapeutic measures intended to cure, slow the progression, reduce the level of symptoms and/or stop the progression of a diagnosed pathological condition or disease, and (2 ) to prophylactic or preventative measures that prevent or slow down the development of a pathological condition or disease (warning or preventing). Thus, those in need of treatment include those who already have the disease, those who are prone to the disease, and those in whom the disease should be prevented.

The terms treat cancer, treat cancer, or their equivalents mean to reduce, reduce, or suppress the replication of cancer cells, reduce, reduce, or suppress the spread (metastasis) of cancer, reduce tumor size, reduce the number of tumors (i.e., reduce tumor burden), reduce or reduce the number of cancer cells in the body, prevent cancer recurrence after surgical removal of a tumor or after other anti-cancer therapies, and/or improve measurable treatment outcome measures (i.e., treatment outcome).

The term standard treatment as used herein is that which is generally accepted in the art and should be understood to refer to the method of treating cancer most widely used in clinical practice. It is usually referred to as chemotherapy, which can be combined with surgery and radiation therapy to treat cancer. Descriptions of specialized treatment regimens can be found on websites maintained by, for example, the National Cancer Institute (www.cancer.gov), the American Society for Clinical Oncology (www.asco.gov). org)] and the US National Comprehensive Cancer Network (www.nccn.org)]. In some embodiments, the term first-line treatment or first-line therapy corresponds to a term generally recognized in the art, which should be understood as the initial treatment of a disease, which is usually part of standard treatment regimens, such as surgery followed by chemotherapy and radiation therapy, which are considered best treatment methods and are also called primary treatment or primary therapy. In some embodiments, the term second-line treatment or second-line therapy corresponds to a term generally recognized in the art, which should be understood as chemotherapy treatment administered when the initial or primary treatment (first-line therapy or primary therapy) has failed or has ceased to work. results and can also be combined with surgery, radiation therapy and/or immunotherapy. As used herein, the term prior treatment may include all types and courses of treatment that a patient received for the treatment of a given disease prior to being prescribed a treatment regimen according to the present invention, including, but not limited to, all first-line, second-line, third-line treatments etc.

The terms synergy, synergistically, synergistically, or mutually enhanced as used herein refer to the effect of interaction or combination of two or more components, resulting in a combined effect greater than the sum of their individual effects (or their additive effect).

The term cancer refers to the phenomenon of the presence of cells having characteristics typical of carcinogenic cells, such as uncontrolled proliferation, cellular immortality, the ability to metastasize, rapid growth and proliferation rate, and certain morphological features. Often cancer cells present as a tumor or mass, but such cells can exist individually within a subject's body or circulate in the bloodstream as independent cells, such as leukemia or lymphoma cells.

The term cancer includes, for example, AIDS-associated cancers, breast cancers, digestive and gastrointestinal cancers, endocrine and neuroendocrine cancers, eye cancers, genitourinary cancers, germ cell cancers, gynecological cancers, head and neck cancers, hematologic cancers, musculoskeletal cancers, neurological cancers, respiratory and thoracic cancers, skin cancers, childhood cancers, and cancers of unknown primary site.

Typical types of AIDS-associated cancers include, but are not limited to, AIDS-associated lymphoma, primary central nervous system lymphoma, and Kaposi's sarcoma.

Typical types of digestive and gastrointestinal cancers include, but are not limited to, anal cancer, anal cancer, appendix cancer, gastrointestinal carcinoid tumor, bile duct cancer, carcinoid tumor, gastrointestinal cancer, colon cancer, esophageal cancer , gallbladder cancer, gastrointestinal stromal tumors (GIST), pancreatic islet tumors, pancreatic neuroendocrine tumors, liver cancer, pancreatic cancer, rectal cancer, colorectal adenocarcinoma, small bowel cancer, gastroesophageal junction (GEJ) cancer , gastric adenocarcinoma and gastric cancer.

Typical types of endocrine and neuroendocrine cancers include, but are not limited to, adrenocortical carcinomas, gastrointestinal carcinoid tumors, pancreatic islet tumors, pancreatic neuroendocrine tumors, adrenocortical carcinoma, Merkel cell carcinomas, non-small cell neuroendocrine lung tumors, small cell neuroendocrine lung tumors, cancers of the parathyroid glands, pheochromocytomas, pituitary tumors and cancers of the thyroid gland.

Common genitourinary cancers include, but are not limited to, bladder cancer, kidney cancer (renal cell carcinoma), penile cancer, prostate cancer, renal pelvis and ureter cancer, transitional cell testicular cancer, urethral cancer, Wilms tumor, and others. pediatric renal tumors.

Common gynecologic cancers include, but are not limited to, cervical cancer, endometrial (uterine) cancer, uterine cancer, fallopian tube cancer, gestational trophoblastic tumor, epithelial ovarian cancer, ovarian germline cell tumor, low-grade (borderline) ovarian tumor. malignancy potential, primary peritoneal cancer, uterine sarcoma, vaginal cancer and vulvar cancer.

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Common head and neck cancers include, but are not limited to, hypopharyngeal cancer, laryngeal cancer, lip and oral cavity cancer, metastatic squamous cell carcinoma of the neck with an occult primary site, oropharyngeal cancer, nasopharyngeal cancer, oral cavity cancer, lip and oropharyngeal cancer, cancer paranasal sinuses and nasal cavity, parathyroid cancer, pharyngeal cancer, salivary gland cancer, throat cancer and thyroid cancer.

Common hematologic cancers include, but are not limited to, leukemias, acute lymphoblastic leukemia, adult and childhood acute lymphoblastic leukemia, adult acute myeloid leukemia, childhood acute myeloid leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, villous leukemia s cells, lymphomas, AIDS-associated lymphoma, cutaneous T-cell lymphoma, Hodgkin's lymphoma in adulthood, Hodgkin's lymphoma in childhood, Hodgkin's lymphoma in pregnancy, fungoid mycoses, non-Hodgkin's lymphoma in adulthood, non-Hodgkin's lymphoma in childhood, non-Hodgkin's lymphoma during pregnancy, primary lymphoma of the central nervous system, Sezary syndrome (reticulosis), cutaneous T-cell lymphoma, Waldenström macroglobulinemia, chronic myeloproliferative neoplasms, Langerhans cell histiocytosis, multiple myeloma or plasma cell neoplasms, myelodysplastic syndromes and myelodysplastic/myeloproliferative neoplasms.

Typical musculoskeletal cancers include, but are not limited to, bone cancer, Ewing's sarcoma, osteosarcoma, malignant fibrous histiocytoma of bone, childhood rhabdomyosarcoma, chondrosarcoma, and soft tissue sarcoma.

Common neurological cancers include, but are not limited to, adult brain tumors, childhood brain tumors, astrocytomas, brain and spinal cord tumors, brainstem glioma, glioblastoma multiforme, atypical teratoid rhabdoid tumor of the central nervous system, embryonal tumors central nervous system, germ cell tumors of the central nervous system, astrocytomas, ependymogliomas, schwannomas, medulloblastomas, meningiomas, craniopharyngiomas, neuroblastomas, pituitary tumors, pituitary adenomas and primary central nervous system (CNS) lymphoma.

Typical respiratory and thoracic cancers include, but are not limited to, non-small cell lung cancer, small cell lung cancer, malignant mesothelioma, thymoma and thymic carcinoma.

Typical skin cancers include, but are not limited to, cutaneous T-cell lymphoma, Kaposi's sarcoma, melanoma, Merkel cell carcinoma, skin cancer, cutaneous T-cell lymphoma, mycosis fungoides, intraocular melanoma, and Sezary syndrome (reticulosis).

Cancers also include resistant versions of all of the cancers listed above or combinations of one or more of the cancers listed above. Some of the cancers given as examples above act as umbrella terms and also as individual terms. For example, genitourinary cancer is an umbrella term and includes bladder cancer, prostate cancer, kidney cancer, testicular cancer, etc., and hepatobiliary cancer, another umbrella term, includes liver cancer (which is itself an umbrella term and includes hepatocellular carcinoma or cholangiocarcinoma), gallbladder cancer, bile duct cancer or pancreatic cancer. Moreover, both of these general terms, namely cancer of the genitourinary system and hepatobiliary cancer, are considered in this description of the invention as separate types of cancer and are included in the general term cancer.

Also included in the term cancer for use herein is the term solid tumor. The term solid tumor refers to conditions such as cancer in which an abnormal tumor mass forms, such as various types of sarcoma, carcinoma and lymphoma. Examples of solid tumors include, but are not limited to, non-small cell lung cancer (NSCLC), neuroendocrine tumors, thymomas, fibrous tumors, metastatic colorectal cancer (mCRC), and the like. In some embodiments, the solid tumor disease is adenocarcinoma, squamous cell carcinoma, large cell carcinoma, or the like.

In some embodiments, the cancer is selected from pancreatic adenocarcinoma, gastric adenocarcinoma, gastroesophageal junction (GEJ) adenocarcinoma, gastroesophageal junction adenocarcinoma, non-small cell lung cancer (NSCLC), breast cancer, triple negative breast cancer (TNBC; i.e. breast cancer testing negative for estrogen receptor (ER-), progesterone receptor (PR-) and HER2 (receptor tyrosine protein kinase erbB-2, also known as CD340 (differentiation cluster 340), Neu proto-oncogene, ERBB2 (human ); HER2-)), ovarian cancer, platinum-resistant ovarian cancer (PROC), melanoma, small cell lung cancer and cholangiocarcinoma. In some embodiments, the cancer is pancreatic ductal adenocarcinoma.

Typical pancreatic neuroendocrine tumors (pancreatic NET or PNET tumors) include, but are not limited to, gastrinomas (Zollinger-Ellison syndromes), glucagonomas, insulinomas, somatostatinomas, VIPomas (Werner-Morrison syndromes), pancreatic

- 8 046190 cholera (WDHA (watery diarrhea + hypokalemia + hypochlorhydria) syndromes), non-functioning pancreatic islet tumors and multiple endocrine neoplasia type 1 (MEN1, also known as Wermer syndromes).

Typical exocrine pancreatic tumors include, but are not limited to, adenocarcinomas, pancreatic ductal adenocarcinomas (PDAC), acinar cell carcinomas, intraductal papillary mucinous tumors (IPMNs), mucinous cystadenocarcinomas, solid pseudopapillary neoplasms, and pancreatoblastomas.

In some embodiments, the cancers are non-operable, advanced, resistant, recurrent or metastatic cancers.

The terms progress, progressive, and progressing as used herein refer to at least one of the following: (1) response to prior therapy (eg, chemotherapy) of progressive disease (PD), (2) appearance of one or more lesions following treatment modalities pretherapy (eg, chemotherapy), (3) to at least a 5% (eg, 10 or 20%) increase in the sum of target lesion diameters relative to the smallest sum recorded in the study (this could be the initial initial total diameter, if it was the smallest in this study), and (4) to the obvious development of non-target lesions.

As used herein, the term sensitization or related terms (eg, sensitize or sensitize) means to make subjects previously resistant, unresponsive, or very unresponsive to a treatment regimen (eg, conventional chemotherapy, targeted therapy, or immunotherapy) more sensitive, fully responsive, or more responsive to the same treatment regimen. In some embodiments, the term sensitization or related terms include the concept of resensitize or its equivalents, meaning to make subjects previously resistant, unresponsive, or very unresponsive to a treatment regimen (eg, conventional chemotherapy, targeted therapy, or immunotherapy) as a result of previous exposure of this therapy regimen, again sensitive, fully responsive, or more responsive to the same therapy regimen.

As used herein, the term at least one compound of formula (I) means a compound selected from the group of compounds of formula (I) (I) prodrugs, derivatives, pharmaceutically acceptable salts of any of the foregoing compounds and solvates of any of the above compounds.

The term compound of formula (I) means a compound selected from compounds of formula (I) o

o (|>

or pharmaceutically acceptable salts of any of the above compounds and solvates of any of the above compounds.

In some embodiments, prodrugs or derivatives of compounds having formula (I) are inhibitors of the STAT3 signaling protein. Non-limiting examples of prodrugs of compounds having formula (I) are their phosphoric acid ester and phosphoric acid diester described in patent application (PCT) WO 2009/036099 as compounds number 4011 and 4012, as well as suitable compounds described in US patent No. 9150530 Examples, but not limited to the following, of derivatives of compounds having formula (I) include those disclosed in Patent Application (PCT) WO 2009/036059. Description of Patent Applications (PCT) WO 2009/036099, WO 2009/036059 and US Patent No. 9150530 incorporated herein by reference in its entirety and for all purposes.

The compound having formula (I) and presented below may also be known as 2-acetylnaphtho[2,3-b]furan-4,9-dione, napabukasin and include tautomers thereof.

- 9 046190

Suitable methods for the preparation of 2-acetylnaphtho[2,3-b]furan-4,9-dione (including its crystalline forms and additional cancer cell stemness inhibitors) are described in PCT patent applications WO 2009/036099, WO 2009/036101, WO 2011 /116398, WO 2011/116399 and WO 2014/169078, the contents of each of these applications being incorporated herein by reference in their entirety and for all purposes.

As used herein, the term at least one gemcitabine means a compound selected from the group consisting of gemcitabine, prodrugs, derivatives, pharmaceutically acceptable salts of any of the foregoing compounds and solvates of any of the foregoing compounds. In some embodiments, the term gemcitabine means a compound selected from the group consisting of gemcitabine, pharmaceutically acceptable salts and solvates thereof.

As used herein, the term at least one paclitaxel means a compound selected from the group consisting of paclitaxel, prodrugs, derivatives, pharmaceutically acceptable salts of any of the foregoing compounds and solvates of any of the foregoing compounds. In some embodiments, the term paclitaxel means a compound selected from the group consisting of paclitaxel, pharmaceutically acceptable salts and solvates thereof. In some embodiments, the term paclitaxel refers to a paclitaxel injection approved for manufacture by the China Food and Drug Administration (formerly the China Food and Drug Administration, now the National Medical Products Administration).

As used herein, the compound of formula (I), paclitaxel and gemcitabine, included in the term drug combination, may be mixed together to form a single formulation for administration, or used for administration separately, and these active components may be administered simultaneously , in series, in parallel and separately.

As used herein, the term salt(s) includes acidic and/or basic salts formed with inorganic and/or organic acids and bases. As used herein, the term pharmaceutically acceptable salt refers to those salts which, within the bounds of sound medical judgment, are suitable for use in contact with the tissues of subjects without exhibiting toxicity, irritation, allergic reactions and/or other adverse effects, and comply reasonable balance of harm and benefit. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. various pharmaceutically acceptable salts are described in detail in J. Pharmaceutical Sciences (1977), 66:1-19.

Pharmaceutically acceptable salts can be formed with inorganic or organic acids. Non-limiting examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid. Non-limiting examples of suitable organic acids include acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid and malonic acid. Other non-limiting examples of suitable pharmaceutically acceptable salts include adipates, alginates, ascorbates, aspartates, benzenesulfonates, besilates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentane propionates, digluconates, dodecyl sulfates, ethanesulfonates, formates, fumarates, glucoheptonates, glycol erophosphates, gluconates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxyethanesulfonates, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalene sulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates and valerates.

In some embodiments, organic acids from which salts can be formed include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, lactic acid, trifluoroacetic acid, maleic acid, malonic acid, succinic acid, fumaric acid. acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid and salicylic acid.

Salts can be prepared in situ during the preparation and purification of the disclosed compound or separately, for example, by reacting said compound with a suitable base or acid, respectively. Non-limiting examples of pharmaceutically acceptable salts derived from bases include alkali metal salts, alkaline earth metal salts, ammonium salts (NH ₄ ⁺ ) and quaternary ammonium base salts (Malkyl Cj-C ₄ ) ₄ ⁺ ). Non-limiting examples of suitable alkali and alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese and aluminum salts. Other non-limiting examples of suitable pharmaceutically acceptable salts include, as appropriate, non-toxic ammonium salts, quaternary ammonium base salts, and amine cation salts formed with counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower ions. alkylsulfonates and arylsulfonates. Non-limiting examples of suitable organic bases from which salts can be prepared include primary amines, secondary amines,

- 10 046190 natural amines, substituted amines, including natural substituted amines, cyclic amines and basic ion exchange resins such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine and ethanolamine. In some embodiments, pharmaceutically acceptable base addition salts such as ammonium, potassium, sodium, calcium and magnesium salts may be selected.

The term solvate is a complex comprising one or more molecules of a compound as described herein and one or more molecules of a solvent or solvents. Solvates of the compounds herein include, for example, hydrates.

The term low dose as used herein refers to a therapeutically effective amount of a drug that is below its standard therapeutic dose. For example, the standard therapeutic dose of gemcitabine is 1000 to 1200 mg/m ² . In some embodiments, a low dose of gemcitabine means less than about 800 mg/m ² , about 600 mg/m ² , about 550 mg/m ² , about 500 mg/m ² , about 450 mg/m ² , about 400 mg/m ² , about 350 mg/m ² , about 300 mg/m ² , about 250 mg/m ² , about 200 mg/m ² , about 150 mg/m ² or about 100 mg/m ² administered weekly. In some embodiments, the low dose of gemcitabine is less than 50% of the recommended effective dose or even lower.

In some embodiments, gemcitabine and paclitaxel are administered according to the therapeutic regimen on days 1, 8, and 15 of each 28-day cycle. In some embodiments, a low dose of gemcitabine (i.e., about 600 mg/ ^m2 , about 550 mg/ ^m2 , about 500 mg/ ^m2 , about 450 mg/ ^m2 , about 400 mg/ ^m2 , about 350 mg /m ² , about 300 mg/m ² , about 250 mg/m ² , about 200 mg/m ² , about 150 mg/m ² or about 100 mg/m ² ) is administered weekly.

In some embodiments, a low dose of gemcitabine (i.e., about 600 mg/ ^m2 , about 550 mg/ ^m2 , about 500 mg/ ^m2 , about 450 mg/ ^m2 , about 400 mg/ ^m2 , about 350 mg /m ² , about 300 mg/m ² , about 250 mg/m ² , about 200 mg/m ² , about 150 mg/m ² or about 100 mg/m ² ) administered weekly for up to 7 weeks.

In some embodiments, a low dose of gemcitabine (i.e., about 600 mg/ ^m2 , about 550 mg/ ^m2 , about 500 mg/ ^m2 , about 450 mg/ ^m2 , about 400 mg/ ^m2 , about 350 mg /m ² , about 300 mg/m ² , about 250 mg/m ² , about 200 mg/m ² , about 150 mg/m ² or about 100 mg/m ² ) administered weekly for 3 of every 4 weeks.

In some embodiments, a low dose of gemcitabine from 100 to 800 mg/m ² is administered weekly, with a low dose of 100 to 600 mg/m ² being more preferred. In some embodiments, a low dose of gemcitabine means about 800 mg/m ² , about 600 mg/m ² , about 300 mg/m ² , or about 100 mg/m ² . In some embodiments, a low dose of gemcitabine of less than about 600 mg/m ² is administered weekly.

In some embodiments, paclitaxel at a dose of about 100 mg/ ^m2 , about 80 mg/ ^m2 , about 70 mg/ ^m2 , about 60 mg/ ^m2 , about 50 mg/ ^m2 , about 40 mg/ ^m2 , about 30 mg/m ² , about 20 mg/m ² or about 10 mg/m ² is administered weekly.

In some embodiments, paclitaxel at a dose of about 100 mg/ ^m2 , about 80 mg/ ^m2 , about 70 mg/ ^m2 , about 60 mg/ ^m2 , about 50 mg/ ^m2 , about 40 mg/ ^m2 , about 30 mg/m ² , about 20 mg/m ² or about 10 mg/m ² is administered weekly for up to 7 weeks.

In some embodiments, paclitaxel at a dose of about 100 mg/ ^m2 , about 80 mg/ ^m2 , about 70 mg/ ^m2 , about 60 mg/ ^m2 , about 50 mg/ ^m2 , about 40 mg/ ^m2 , about 30 mg/m ² , about 20 mg/m ² or about 10 mg/m ² is administered weekly for 3 of every 4 weeks.

In some embodiments, paclitaxel at a dose of 10 to 100 mg/m ² is administered weekly, with a dose of 40 to 80 mg/m ² being more preferred. In some embodiments, paclitaxel at doses of about 80 mg/m ² , about 60 mg/m ² or about 40 mg/m ² is administered weekly. In some embodiments, paclitaxel at a dose of less than 80 mg/m ² is administered weekly. In some embodiments, paclitaxel is administered intravenously at a dose of 80 mg/m ² according to a predetermined regimen, starting at least 2 hours after administration of the compound of formula (I) on the same day, with the intravenous infusion continued for approximately 60 minutes. In some embodiments, gemcitabine is administered intravenously at a dose of 600 mg/m ² immediately after intravenous administration of paclitaxel according to a predetermined treatment regimen, with intravenous administration of gemcitabine continued for about 30 to 60 minutes.

In some embodiments, a compound of formula (I) can be administered at a total daily dose ranging from about 80 to about 960 mg. In some embodiments, a compound of formula (I) can be administered at a total daily dose ranging from about 80 to about 480 mg. In some embodiments, a compound of formula (I) can be administered at a total daily dose ranging from about 160 mg to about 480 mg. In some embodiments, the total amount of the compound of formula (I) is administered once daily. In some embodiments, the compound of Formula (I) is administered at a dose of about 480 mg. In some embodiments, the compound of Formula (I) is administered at a dose of about 400 mg. In some embodiments, the compound of Formula (I) is administered at a dose of about 320 mg. In some embodiments, the compound of Formula (I) is administered at a dose of about 240 mg. In some embodiments, a compound of formula (I) is administered

- 11 046190 at a dose of approximately 160 mg. In some embodiments, the total amount of a compound of Formula (I) is administered as a divided total dose (more than one time) over a single day, for example, twice daily (BID) or more frequently. In some embodiments, a compound of formula (I) can be administered at a total daily dose ranging from about 80 to about 480 mg, twice daily. In some embodiments, a compound of formula (I) can be administered at a total daily dose ranging from about 80 to about 240 mg, twice daily. In some embodiments, a compound of formula (I) is administered at a total daily dose of about 240 mg twice daily. In some embodiments, a compound of formula (I) is administered at a total daily dose of about 480 mg twice daily. In some embodiments, a compound of Formula (I) is administered at a total daily dose of about 160 mg twice daily. In some embodiments, a compound of formula (I) is administered at a total daily dose of about 80 mg twice daily. In some embodiments, a compound of formula (I) is administered at a dose of about 240 mg twice daily at intervals of 8 to 12 hours. In some embodiments, a compound of formula (I) is administered at a dose of about 80 mg twice daily at intervals of 8 to 12 hours. until 12 o'clock

The compound disclosed herein may be in the form of a pharmaceutical composition (pharmaceutical preparation). In some embodiments, the pharmaceutical compositions (pharmaceuticals) may include a compound of formula (I) and at least one pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical compositions (pharmaceuticals) may include a compound of formula (I), paclitaxel, and a low dose of gemcitabine. In some embodiments, the pharmaceutical compositions (pharmaceuticals) may include a compound of formula (I) and paclitaxel. In some embodiments, the pharmaceutical compositions (pharmaceuticals) may include a compound of formula (I) and a low dose of gemcitabine.

The term carrier, as used herein, means a pharmaceutically acceptable material, composition or transport substance, such as a liquid or solid excipient, diluent, excipient, solvent or coating material, which is capable of somehow carrying or transporting the pharmaceutical compound in question from one organ or part body to another organ or part of the body. Each carrier must be acceptable in the sense of its compatibility with the other components of the formulation and not be harmful to the patient. Non-limiting examples of pharmaceutically acceptable carriers and/or diluents include sugars such as lactose, glucose and sucrose, starches such as corn starch and potato starch, cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate, tragacanth powder, malt , gelatin, talc, excipients such as cocoa butter and suppository waxes, vegetable oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil, glycols such as propylene glycol, polyhydric alcohols (polyols) such as glycerin, sorbitol, mannitol and polyethylene glycol, esters such as ethyl oleate and ethyl laurate, agar, buffering agents such as magnesium hydroxide and aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, phosphate buffer solutions and other non-toxic and compatible substances commonly used in pharmaceutical formulations. The compositions may also contain various humectants, emulsifiers and lubricants, such as sodium lauryl sulfate, magnesium stearate and polyethylene oxide-polypropylene oxide copolymer, as well as color additives, release agents, film formers, sweeteners, flavoring agents, preservatives and antioxidants.

The pharmaceutical compositions (pharmaceutical preparations) disclosed herein, which are suitable for oral administration, may be presented in the form of capsules, cachets, pills, tablets, lozenges (using a flavor base, usually sucrose and gum arabic or tragacanth), powders, granules, dragees, solution in an aqueous or non-aqueous liquid, suspension in an aqueous or non-aqueous liquid, oil-in-water emulsion, water-in-oil emulsion, tincture, syrup, lozenges (using an inert base such as gelatin, glycerin, sucrose and/or or gum acacia) and/or mouthwashes, each such form would contain a predetermined amount of the compound disclosed herein.

The pharmaceutical composition (pharmaceutical preparation) described herein can be administered in the form of boluses, electuaries or pastes.

Solid dosage forms for oral administration (capsules, tablets, pills, dragees, powders, granules, etc.) can be formulated in admixture with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following components: fillers or dry diluents such as starches, lactose, sucrose, glucose, mannitol and/or silicic acid, binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or acacia, humectants, such as glycerin, disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, some silicates, sodium carbonate and sodium starch

- 12 046190 glycolate, dissolution retarders such as paraffin, absorption accelerators such as quaternary ammonium compounds, wetting agents such as, for example, cetyl alcohol, glycerol monostearate and polyethylene oxide-polypropylene oxide copolymer, absorbents such as kaolin and bentonite clay, lubricants additives such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof, and color additives. In the case of capsules, tablets and pills, the pharmaceutical compositions may also include buffering agents. Solid compositions of a similar type may also be used as fillers for soft or hard gelatin capsules using excipients such as lactose or milk sugars, high molecular weight polyethylene glycols and the like.

Liquid dosage forms for oral administration may include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and tinctures. In addition to the active ingredients, liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol , 1,3-butylene glycol, vegetable oils (especially cottonseed, peanut, corn, wheat germ, olive, castor or sesame oil), glycerin, tetrahydrofuryl alcohol, polyethylene glycols and sorbitol fatty acid esters, as well as mixtures thereof. In addition, cyclodextrins, such as hydroxypropyl-beta-cyclodextrin, can be used to solubilize these substances.

Pharmaceutical compositions (pharmaceutical preparations) may also include auxiliary medicinal substances (adjuvants), such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, coloring agents, flavoring agents and preservatives. Suspensions, in addition to the substances disclosed herein, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitol esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, as well as mixtures thereof.

The pharmaceutical compositions disclosed herein for rectal or vaginal administration may be in the form of suppositories, which may be prepared by mixing one or more of the components herein with one or more suitable non-irritating excipients or carriers, including, for example, coconut oil, polyethylene glycol, suppository wax or salicylate, which are solid at room temperature but liquid at human body temperature and therefore melt within the rectum or vaginal cavity, releasing the substances disclosed herein. Pharmaceutical compositions that are suitable for vaginal administration may also include pessaries, tampons, creams, gels, pastes, foams or aerosol spray formulations containing carriers known to be suitable in the art.

Dosage forms for topical and transdermal administration of a pharmaceutical composition (pharmaceutical preparation) or pharmaceutical tablet according to the present description may include powders, aerosols (sprays), ointments, pastes, creams, lotions, gels, solutions, patches and inhalers. The pharmaceutical composition or pharmaceutical tablet may be mixed under sterile conditions with a pharmaceutically acceptable carrier and any preservatives, buffers or propellants that may be required.

Ointments, pastes, creams and gels may contain, in addition to the pharmaceutical composition (pharmaceutical preparation) or pharmaceutical tablet according to the present description, excipients such as animal and vegetable fats and oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones , bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and aerosols (sprays) may contain, in addition to the pharmaceutical composition (pharmaceutical preparation) or pharmaceutical tablet according to the present description, also excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and powdered polyamides, or mixtures thereof. In addition, sprays may contain conventional propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

Ophthalmic compositions, eye ointments, powders, solutions, etc. are also considered dosage forms within the scope of the present invention.

Compositions (pharmaceutical preparations) suitable for parenteral administration may include at least one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted directly into sterile injectable solutions or dispersions before their use, they may contain antioxidants, buffers, bacteriostats, soluble additives to provide the formulation with isotonic properties relative to the blood composition of the target recipient, or suspending or thickening agents.

In various embodiments of the invention, the compound of formula (I) described herein

- 13 046190 document, contained in pharmaceutical compositions. Said pharmaceutical compositions (pharmaceutical preparations) include the compounds of formula (I), their pharmaceutically acceptable salts and solvates, and one or more surfactants. In some embodiments, the surfactant is sodium lauryl sulfate (SLS), sodium dodecyl sulfate (SDS), or one or more polyoxyglycerides. For example, lauryl polyoxyglycerides (sometimes referred to as Gelucire™) or linoleoyl polyoxyglycerides (sometimes referred to as Labrafil™) can be used as the polyoxyglyceride. Examples of such compositions are presented in PCT patent application No. WO 2014/169078, the contents of which are included in this description in its entirety.

At least four properties of CSC cells are considered to contribute to their malignancy. These are their stemness, dysregulation of stemness signaling pathways, resistance to traditional cancer treatments and tendency to metastasize. As used herein, the term stemness generally refers to the ability of a population of stem cells to self-renew and transform into cancer stem cells (CSC) (Gupta P.B. et al., Nat. Med. 2009; 15(9):1010-1012). Although CSC cells constitute only a small percentage of the total population of cancer cells in a tumor (Clarke M.F., Biol. Blood Marrow Transplant. 2009; 11 (2 suppl. 2):14-16), they give rise to heterogeneous lineages of differentiated cancer cells that make up the main part of the tumor (see Gupta et al., 2009). In addition, CSC cells have the ability to spread to other sites in the body through metastasis, where they provide seeds for the growth of new tumors (Jordan C.T. et al. N. Engl. J. Med. 2006; 355(12):1253-1261).

The initiation and maintenance of stemness properties in CSC cells is mediated by progressive dysregulation of stemness signaling pathways, including, but not limited to, the Janus kinase/signal transporters and activators of transcription (JAK/STAT) associated signaling pathways, namely Hedgehog (Desert (DHH) , Indian (IHH) and Sonic (SHH))/PATCHED/(PTCH1)/SMOOTHENED (SMO), NOTCH/DELTA-LIKE (DLL1, DLL3, DLL4)/JAGGED (JAG1, JAG2)/CSL (CBF1/Su(H )/Lag-1), WNT/APC/GSK3/p-CATENIN/TCF4 and NANOG (Boman B.M. et al., J. Clin. Oncol. 2008; 26(17):2828-2838).

It is the dysregulation of these stemness signaling pathways in CSC cells (see Boman et al. 2008) that is considered to be the factor that makes CSC cells resistant to chemotherapy and radiation therapy, which subsequently leads to the recovery and further spread of cancer. Thus, although chemotherapy and radiation therapy kill most rapidly dividing cancer cells in the bulk of the tumor, dysregulation of stemness signaling pathways in CSC cells may allow CSC cells to escape cell death from chemotherapy and also explain how surviving CSC cells acquire the ability to metastasize to sites in the body , sufficiently distant from the primary tumor site.

Signal Transducer and Activator of Transcription 3 or STAT3, also known as Acute-Phase Response Factor or APRF, DNA-Binding Protein APRF, ADMIO 3, HIES, functions at the interface of several cytokine signaling pathways (see for example, Fig. 3, Catlett-Falcone, R., et al. Immunity, 1999. 10(1): p. 105-15; Bromberg, J. F., et al. Cell, 1999. 98(3): p. 295 -303; Kanda, N., et al. Oncogene, 2004. 23(28): p. 4921-29; Schlette, E. J., et al. J. Clin Oncol, 2004. 22(9): p. 1682-88 ; Niu, G., et al. Oncogene, 2002. 21(13): p. 2000-08; Xie, T. X., et al. Oncogene, 2004. 23(20): p. 3550-60). STAT3 regulates not only the expression of genes that control the cell cycle (CYCLIN D1, D2 and c-MYC), cell survival (BCL-XL, BCL-2, MCL-1) and angiogenesis (HIF1a, VEGF) (Furqan et al. Journal of Hematology & Oncology (2013), 6:90), but is also a major negative regulator of tumor immune surveillance and immune cell mobilization at the lesion site (Kortylewski, M., et al. Nat. Med., 2005. 11(12) : p. 1314-21; Burdelya, L., et al. J. Immunol., 2005. 174(7): p. 3925-31 and Wang, T., et al. Nat. Med., 2004. 10( 1): p. 48-54).

In normal cells, STAT3 activation is transient and tightly regulated, lasting, for example, from about 30 minutes to several hours. However, in a wide variety of human cancers, including all major carcinomas as well as some hematologic tumors, STAT3 appears abnormally active (Lin et al., Oncogene (2000), 19, 2496-2504; Bromberg J. Clin. Invest. (2002 ), 109:1139-1142; Buettner et al., Clinical Cancer Research (2002), 8, 945-954; Frank Cancer Letters, 251 (2007), 199-21; Oyu et al. Nature Reviews Cancer (2004), 4, 97-105). Consistently active STAT3 is present in more than half of all breast and lung cancers, as well as colorectal cancers (CRC), ovarian cancers, hepatocellular carcinomas, multiple myelomas, pancreatic cancers and more than 95% of all cancers head and neck.

Suppression of STAT3 signaling function through antisense oligonucleotides, short interfering RNA (siRNA), dominant negative form of STAT3 and/or targeted inhibition of STAT3-dependent tyrosine kinase activity leads to inhibition of cancer cell growth, apoptosis and a decrease in the incidence of metastasis in vitro and/or in vivo, assuming that the trunk

- 14 046190 CSC cell activity is dependent on the constitutive activation of the transcription factor STAT3 (Pedranzini, L., et al. J. Clin. Invest., 2004. 114(5): p. 619-22; Bromberg, J.F., et al Cell 1999 98(3): 295-303 Darnell JE Nat Med 2005 11(6): 595-96 and Zhang L et al Cancer Res 2007 67(12): pp. 5859-64). Therefore, STAT3 may play a key role in the ability of CSC cells to survive and self-renew in a wide range of cancer types.

As mentioned above, the methods disclosed herein can treat at least one disease associated with impaired STAT3 signaling activity in a subject. Abnormal activity of the STAT3 signaling pathway can be identified by the expression of phosphorylated STAT3 protein (pSTAT3) or its surrogate upstream or downstream regulators, or by detection of pSTAT3 localized in the nucleus.

The STAT3 signaling pathway can be activated in response to cytokines, such as IL-6, or one or more tyrosine kinases, such as EGFR, JAKs, ABL, KDR, c-MET, SRC and HER2 (see, for example, Fig. 3). Downstream effectors of STAT3 include, but are not limited to, BCL-XL, c-MYC, CYCLIND1, VEGF, MMP-2, and SURVIVIN. The STAT3 signaling pathway has been found to be abnormally active in a wide range of cancers. The persistently active STAT3 signaling pathway may be present in more than half of all breast and lung cancers, hepatocellular carcinomas, multiple myelomas, and more than 95% of all head and neck cancers. Blocking the STAT3 signaling pathway leads to inhibition of cancer cell growth, apoptosis, and a decrease in the frequency of metastasis in vitro and/or in vivo. Activated STAT3 has also been demonstrated in a number of autoimmune and inflammatory diseases. In addition, since it has been discovered that interleukin-6-mediated inflammation is the most common cause for the onset of atherosclerosis, peripheral blood vessel disease, coronary heart disease, hypertension, osteoporosis, type 2 diabetes and dementia, and it has also been discovered Since gp130-JAKS-STATs is a major signaling pathway activated by IL-6, inhibition of STAT3 signaling may also help treat or prevent these diseases (Libby, P., et al. Circulation, 2002. 105(9): p. 1 135-43; Stephens, J. W., et al. Mol. Genet. Metab., 2004. 82(2): p. 180-86; Cesari, M., et al. Circulation, 2003. 108(19) : p. 2317-22; Orshal, J. M. and R. A. Khalil. Am. J. Physiol. Regul. Integr. Comp. Physiol., 2004. 286(6): p. R1013-23; Manolagas, S. S. Bone, 1995. 17(2 SuppI): pp. 63S-67S and Yaffe, K., et al. Neurology, 2003. 61(1): pp. 76-80).

In some embodiments, the at least one disease may be selected from cancers having abnormal STAT3 signaling pathway activity. For example, activated pSTAT3 has been found in pancreatic cancer cells (Wei et al. Oncogene (2003), 22(3):319-329; Scholz et al. Gastroenterology (2003), 125:891-905; Toyonaga et al. Cancer Lett (2003), 10;201(1):107-16;Qiu et al. Cancer Sci. (2007), 98(7):1099-106).

In some embodiments, the at least one disease may be selected from autoimmune diseases associated with abnormal activity of the STAT3 signaling pathway and inflammatory diseases associated with abnormal activity of the STAT3 signaling pathway. In some embodiments, diseases associated with abnormal STAT3 signaling pathway activity may be selected from inflammatory bowel disease, arthritis, Crohn's disease, ulcerative colitis, rheumatoid arthritis, asthma, allergy, and systemic lupus erythematosus.

In some embodiments, the at least one disease may be selected from CNS diseases associated with abnormal activity of the STAT3 signaling pathway. In some embodiments, the CNS diseases may be selected from autoimmune diseases associated with demyelination, Alzheimer's disease, stroke, ischemia and reperfusion injury, and multiple sclerosis. In some embodiments, the at least one disease may be selected from diseases caused by inflammation and associated with abnormal activity of the STAT3 signaling pathway. In some embodiments, inflammatory diseases associated with abnormal activity of the STAT3 signaling pathway may be selected from peripheral vascular disease, coronary heart disease, hypertension, osteoporosis, type 2 diabetes, and dementia.

As discussed above, CSC cells are a subpopulation of cancer cells (found within solid tumors or in the bloodstream in hematologic cancers) that exhibit characteristics typically associated with stem cells. These cells can grow faster after the number of non-stem, regular cancer cells is reduced by chemotherapy, and this property may be responsible for the frequent recurrence of cancer after chemotherapy. In contrast to the bulk of cancer cells, which do not produce tumors, CSC cells are tumor-forming (initiating new tumors). In human acute myeloid leukemia, the incidence of these cells is less than 1 in 10,000 cancer cells (Bonnet, D. and J.E. Dick. Nat. Med., 1997. 3(7): p. 730-37).

The effectiveness of various cancer treatments (especially in the early stages of their trials) is often measured by the number and size of tumor mass destroyed by these methods. Since CSC cells constitute a very small proportion of the total tumor cell population and have a markedly

- 15 046190 different biological characteristics than their progeny differentiated cells, measuring tumor mass may be meaningless in the case of the use of drugs that act selectively only on cancer stem cells. In fact, CSC cells are resistant to radiation therapy and resistant to chemotherapeutic agents and targeted drugs. Normal human somatic stem cells are usually inherently resistant to chemotherapeutic agents because they have various protein pumps (eg, multidrug resistance protein pump) that remove drugs, a higher DNA repair capacity, and a slower rate of cell population turnover (chemotherapeutic agents). inherently target rapidly replicating cells). CSC cells, being mutated analogues of normal stem cells, may also have similar properties that allow them to survive therapy. In other words, traditional chemotherapy methods kill differentiated (or differentiating) cells that form the bulk of the tumor, which are unable to generate new cells (see, for example, Fig. 2). The population of CSC cells that gives rise to the tumor may remain unaffected and subsequently cause disease recurrence. In addition, treatment with chemotherapeutic agents may leave behind only chemotherapy-resistant CSC cells, increasing the likelihood that subsequent tumors will also be chemotherapy-resistant. In addition, it has also been demonstrated that cancer stem cells are resistant to radiotherapy (XRT) (Hambardzumyan, et al. Cancer Cell, 2006. 10(6): p. 454-56 and Baumann, M., et al. Nat Rev. Cancer, 2008. 8(7): pp. 545-54).

Because surviving CSC cells can repopulate tumor cells and induce relapse, anticancer therapies that include specific strategies against CSC cells hold great promise (Jones R. J. et al., J. Natl. Cancer Inst. 2004; 96(8):583-585 ). By targeting CSC signaling pathways, it is possible to treat patients with aggressive, unresectable tumors or resistant, recurrent cancers, as well as prevent metastasis and tumor regeneration. The development of specialized therapies targeting CSC signaling pathways is underway. Therefore, an increase in survival time and an improvement in the quality of life of cancer patients, especially those suffering from metastatic disease, can be expected.

Recent studies have discovered that cancer stem cells are capable of regenerating tumors (eg, Fig. 1). These cancer stem cells have been found to be functionally associated with ongoing malignancies, cancer metastasis, cancer relapse, and resistance to anticancer drugs. Cancer stem cells and their progeny differentiated cells have markedly different biological characteristics. Cancer stem cells persist in tumors as well-defined but rare populations. Comparative analyzes of traditional anticancer drugs are based on measurements of tumor number and weight and, therefore, cannot correctly identify those drugs that specifically target cancer stem cells. In fact, it has been discovered that cancer stem cells are resistant to standard chemotherapy and increase in concentration after courses of standard chemotherapy (see, for example, Fig. 2), which can lead to recurrent and chemotherapy-resistant cancer. Cancer stem cells have also been shown to be resistant to radiation therapy (Baumann, M., et al. Nat. Rev. Cancer, 2008. 8(7): p. 545-54). Types of cancer in which cancer stem cells have been isolated from the literature include breast cancer, head cancer, neck cancer, lung cancer, ovarian cancer, pancreatic cancer, colorectal carcinoma, prostate cancer, melanoma, multiple myeloma, sarcoma Kaposi's sarcoma, Ewing's sarcoma, liver cancer, medulloblastoma, brain tumors and leukemia. STAT3 has been identified as a survival and self-renewal factor for cancer stem cells. Therefore, STAT3 inhibitors may kill cancer stem cells and/or may inhibit cancer stem cell self-renewal. In some embodiments, the term cancer stem cells as used herein refers to a very small population of cancer cells that have the ability to self-renew and are tumor-forming (capable of giving rise to new tumors). Previous studies have proven that the compound of formula (I) can effectively inhibit STAT3 signaling pathways, and thereby inhibit the growth and survival of CSC cells. In clinical studies, the compound of formula (I) can be used to treat patients with advanced cancer. Based on previous experiments, the present invention has unexpectedly discovered that new types of pharmaceutical compositions containing a compound of formula (I), low dose gemcitabine and paclitaxel can also effectively treat patients with advanced cancer, especially patients with metastatic pancreatic cancer after unsuccessful previous courses of treatment. Previously, it had never been reported in the literature that a combination chemotherapy regimen using low-dose gemcitabine and paclitaxel could be effectively used to treat patients with advanced pancreatic cancer. Therefore, the present invention provides a new option for improving the survival of patients with advanced metastatic pancreatic cancer, especially after failure of first-line treatment.

This disclosure discloses methods for suppressing, reducing, and minimizing the survival of

- 16 046190 growth and self-renewal of cancer stem cells, including the administration of therapeutically effective pharmaceutical compositions that contain a compound of formula (I) in combination with a low dose of gemcitabine. Disclosed herein are methods of inhibiting, reducing, and minimizing the survival and self-renewal of cancer stem cells, comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a therapeutically effective amount of paclitaxel. Also disclosed herein are methods of inhibiting, reducing, and minimizing the survival and self-renewal of cancer stem cells, comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a low dose of gemcitabine and a therapeutically effective amount of paclitaxel. In some embodiments, a compound of formula (I) is included in a pharmaceutical composition.

Disclosed herein are methods of treating at least one type of cancer resistant to conventional chemotherapy and/or targeted therapy in a subject, comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a low dose of gemcitabine. Also disclosed herein are methods of treating at least one type of cancer resistant to conventional chemotherapy and/or targeted therapy in a subject, comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a therapeutically effective amount or a low dose of gemcitabine on a weekly basis. Also disclosed herein are methods of treating at least one type of cancer resistant to conventional chemotherapy and/or targeted therapy in a subject, comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a low dose of gemcitabine and a therapeutically effective amount of paclitaxel weekly. In various embodiments, a compound of formula (I) is included in a pharmaceutical composition.

Disclosed herein are methods of treating or preventing recurrence of cancer or metastasis thereof in a subject, comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a low dose of gemcitabine. Also disclosed herein are methods of treating or preventing recurrence of cancer or metastasis thereof in a subject, comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a therapeutically effective amount or a low dose of gemcitabine on a weekly basis. Also disclosed herein are methods of treating or preventing recurrence of cancer or metastasis thereof in a subject, comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a low dose of gemcitabine and a therapeutically effective amount or low dose of paclitaxel weekly. In various embodiments, a compound of formula (I) is included in a pharmaceutical composition. In various embodiments, the subject is a pancreatic cancer patient with previous treatment failure. In various embodiments, the subject is a patient with metastatic pancreatic cancer after a previous treatment failure.

Disclosed herein are methods of treating cancer in a subject, comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a low dose of gemcitabine. Disclosed herein are methods of treating cancer in a subject comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a therapeutically effective amount or a low dose of paclitaxel on a weekly basis. Disclosed herein are methods of treating cancer in a subject comprising administering a therapeutically effective amount of a compound of formula (I) in combination with a low dose of gemcitabine and a therapeutically effective amount or low dose of paclitaxel on a weekly basis. In various embodiments, a compound of formula (I) is included in a pharmaceutical composition.

Disclosed herein are the uses of pharmaceutical compositions in methods of treating cancer in a subject, the composition comprising a therapeutically effective amount of a compound of formula (I), a low dose of gemcitabine, and a therapeutically effective amount or low dose of paclitaxel, administered weekly. The compound of formula (I) is administered daily at a dose of 80 to 960 mg. Paclitaxel can be administered at a dose of 10 to 100 mg/ ^m2 daily. Gemcitabine can be administered at a dose of 100 to 800 mg/ ^m2 daily. In some preferred embodiments, the compound of formula (I) is administered at a total daily dose ranging from about 80 to about 480 mg. In some preferred embodiments, paclitaxel is administered at a dose of 40 to 80 mg/m ² weekly. In some preferred embodiments, gemcitabine is administered at a dose of 100 to 600 mg/m ² weekly. In some embodiments, the subject is a patient with pancreatic cancer after a previous treatment failure. In various embodiments, the subject is a patient with metastatic pancreatic cancer after a previous treatment failure.

In some embodiments, each of the cancers may be unresectable, advanced, refractory, recurrent, or metastatic. In some embodiments, the cancer may be pancreatic cancer. In some embodiments, the cancer may be pancreatic cancer that has progressed following a previous course of treatment. In some

- 17 046190 embodiments, the pancreatic cancer may be pancreatic adenocarcinoma. In some embodiments, the pancreatic cancer may be pancreatic ductal adenocarcinoma. In some embodiments, the cancer may be metastatic pancreatic cancer. In some embodiments, the cancer may be metastatic pancreatic cancer that has progressed after a previous course of treatment. In some embodiments, cancer may be associated with overexpression of activated pSTAT3. In some embodiments, cancer may be associated with intranuclear localization of β-catenin.

Examples

The following are examples that further illustrate various features of the present invention. The examples also illustrate useful methodology for the practical application of the present invention. These examples do not limit the claimed invention.

The methods disclosed herein involve administering to a subject in need of treatment a pharmaceutical composition comprising a low dose of gemcitabine, paclitaxel and a compound of formula (I) on a weekly basis.

Example 1.

The effects of 2-acetylnaphtho[2,3-b]furan-4,9-dione (i.e. the compound of formula (I)), gemcitabine and a combination of 2-acetylnaphtho[2,3-b]furan-4,9- dione and gemcitabine. Specifically, immunosuppressed mice bearing established human pancreatic adenoma (Panc-1) were treated under pure vehicle control with 2-acetylnaphtho[2,3-b]furan-4,9-dione (100 mg/kg, p.o., two once daily), gemcitabine (Gemzar, 80 mg/kg, IV, q3d) or a combination of 2-acetylnaphtho[2,3-b]furan-4,9-dione and gemcitabine. Tumor size was assessed periodically during treatment. Each data point represents the mean ± SEM (standard error of the mean) of measurements from five tumors. As shown in FIG. 4, although 2-acetylnaphtho[2,3-b]furan-4,9-dione or gemcitabine showed some effects in suppressing tumor growth, their combination in a mouse model showed a significantly greater reduction in tumor growth.

Example 2.

Recording the effects of 2-acetylnaphtho[2,3-b]furan-4,9-dione with or without paclitaxel on cancer stem cells based on cancer stem cell markers in xenograft cancer animal models.

Human cancer cells were implanted subcutaneously in the area of the right side of the abdominal cavity in 5-7 week old nude mice. When tumor size reached 200 mm ³ , they were administered 2-acetylnaphtho[2,3-b]furan-4,9-dione (e.g., 50 mg/kg twice daily by gavage (in groups of n=3)) , paclitaxel or 2-acetylnaphtho[2,3-b]furan-4,9-dione in combination with paclitaxel for the treatment of animals. Tumors were collected 24 hours after the first dose.

The collected tissues were fixed in 3.7% formaldehyde solution in neutral buffer at 4°C overnight. Then they were embedded in paraffin. Sections of approximately 5 microns were made and these sections were fixed on positively charged glass slides. After warming and dewaxing, slides containing tumor or control tissue samples were incubated in 10 mM sodium citrate solution (pH 6.0) for 10 min. After antigen retrieval, primary antibody p-STAT3 (Rabbit, Cell Signaling, 1:100), β-catenin (Mouse, Santa Cruz, 1:400) was used to label samples overnight at 4°C, and then secondary antibody conjugated with Alexa Fluor Fluorescent dye (1:500, Invitrogen). After mounting in media, slides containing DAPI (Invitrogen) with Prolong mounting medium were imaged through the 20× objective of a Zeiss fluorescence microscope and analyzed using Zen software.

As shown in FIG. 5, paclitaxel alone can lead to increased staining of stem cell markers, and 2-acetylnaphtho[2,3-b]furan-4,9-dione alone can significantly reduce the expression of stem cell markers p-STAT3 and β-catenin Therefore, when 2-acetylnaphtho[2,3EDfuran^D-dione is combined with paclitaxel, the expression of stem cell markers decreases to a lower level.

Example 3.

In phase II/III extended open-label international multicenter clinical studies, the effect of the compound of formula (I) was studied, i.e. 2-acetylnaphtho[2,3-b]furan-4,9-dione, with paclitaxel and low dose gemcitabine in patients with advanced pancreatic cancer after previous treatment failures.

These clinical studies observed the antitumor activity and safety of 2-acetylnaphtho[2,3-b]furan-4,9-dione in combination with weekly paclitaxel and low-dose gemcitabine in patients with advanced pancreatic cancer who had failed previous treatments. Images are available showing disease progression in patients who have previously received at least first-line systemic treatment that failed. Previous treatments included gemcitabine-based monotherapy, combination therapy, and/or FOLFIRINOX/mFOLFIRINOX-type chemotherapy.

- 18 046190

Patients listed to receive a 4-week (28-day) treatment cycle received 2-acetylnaphtho[2,3-b]furan-4,9-dione 240 mg orally twice daily (for a total daily dose of 480 mg) in combination with treatment with paclitaxel and low dose gemcitabine. On the first day of the first cycle, treatment began with paclitaxel 80 mg/m ² intravenously, followed by gemcitabine 600 mg/m ² intravenously on days 1, 8, and 15 of each 28-day treatment cycle. In case of manifestations of drug toxicity, the dose could be adjusted (reduced or even canceled).

To evaluate the antitumor activity of 2-acetylnaphtho[2,3-b]furan-4,9-dione in combination with paclitaxel and a low dose of gemcitabine, tumor remission and progression were analyzed according to the RECIST 1.1 standard.

Among patients enrolled with advanced pancreatic cancer, one patient had received prior third-line chemotherapy (following first-line hecitabine + tegafur, second-line gemcitabine, and third-line avelumab) and had disease progression after enrollment . Following treatment in these clinical trials, target lesion shrinkage was 9.1% at first assessment (8 weeks after study randomization) (RECIST score was SD (stable disease)). At the second evaluation (20 weeks after study randomization), the target lesion had decreased by 41%, the RECIST score had reached PR (partial response), and the nontarget lesions had significantly decreased.

One patient had previously received first-line chemotherapy (capecitabine + tegafur) and had disease progression before and after his enrollment. Following treatment in the clinical trial, target lesions were reduced by 25.6% at the first assessment (8 weeks after study randomization) (RECIST score SD (stable disease)), with significant reductions in non-target lesions. Two patients are currently still undergoing treatment.

Adverse events associated with 2-acetylnaphtho[2,3-b]furan-4,9-dione in all patients treated were primarily gastrointestinal side effects (such as diarrhea, constipation, nausea, vomiting etc.), while the combined drug regimen did not increase these side effects in patients and was characterized by good safety.

These studies show that 2-acetylnaphtho[2,3-b]furan-4,9-dione in combination with paclitaxel and low dose gemcitabine has a good synergistic effect in patients with metastatic pancreatic cancer after previous treatment failures. It has been shown that 2-acetylnaphtho[2,3-b]furan-4,9-dione (240 mg twice daily) in combination with paclitaxel (80 mg/ ^m2 intravenously once a week) and gemcitabine (600 mg/m m ² intravenously) is safe, well tolerated and has good antitumor activity.

Many features and advantages of the present invention will become apparent from the detailed description thereof, and it is the intention of the inventors to cover by the appended claims all such features and advantages of the invention described within the true spirit and scope of the present invention. In addition, since a variety of changes and variations can easily be made to the contents of the invention by those skilled in the art, it is desirable not to be limited in this specification to any of the precise structures and operations that would be suitably illustrated and described herein, but rather to include all possible changes and equivalent embodiments may be considered to fall within the scope of the present invention.

Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art when used in the particular manner in which they are used.

Incorporation of literature data by reference

This disclosure contains references and quotations from other literature such as patents, patent applications, patent publications, journals, books, articles, websites, and the like. All of these literature sources are incorporated herein by reference in their entirety and for all purposes. Any material or portion thereof which is stated to be incorporated by reference herein, but which conflicts with existing definitions, provisions, or other specification material expressly set forth in this specification, shall be deemed to be included only to the extent not creates inconsistencies between the material included and the material in this description. If any conflict arises, it shall be resolved in favor of the present disclosure as the preferred alternative.

Claims (10)

1. A method of treating chemotherapy-resistant pancreatic cancer, comprising administering to a subject in need of such treatment: a compound of formula (I) or pharmaceutically acceptable salts or solvates thereof at a dose of from about 480 to about 960 mg per day gemcitabine or its pharmaceutically acceptable salts or solvates at a dose of about 600 mg/m ² once a week by intravenous infusion; paclitaxel or its pharmaceutically acceptable salts or solvates at a dose of approximately 80 mg/m ² once a week by intravenous infusion. 2. The method according to claim 1, wherein pancreatic cancer is resistant to traditional methods of chemotherapy. 3. A method of resensitizing a subject with pancreatic cancer whose previous treatment with chemotherapy has failed, comprising administering to said subject: a compound of formula (I) or pharmaceutically acceptable salts or solvates thereof at a dose of from about 480 to about 960 mg per day gemcitabine or its pharmaceutically acceptable salts or solvates at a dose of about 600 mg/m ² once a week by intravenous infusion; paclitaxel or its pharmaceutically acceptable salts or solvates at a dose of approximately 80 mg/m ² once a week by intravenous infusion. 4. The method according to any one of claims 1 to 3, wherein the cancer is late, metastatic, unresectable, resistant or recurrent, chemotherapy-resistant cancer. 5. The method according to any one of claims 1 to 3, wherein the subject has received at least one previous course of chemotherapy for pancreatic cancer that was unsuccessful. 6. The method according to any one of claims 1 to 3, wherein the subject has developed pancreatic cancer to the stage of metastatic cancer after at least one previous course of treatment. 7. The method according to any one of claims 1 to 3, wherein the compound of formula (I), gemcitabine and paclitaxel can be used in second-line treatment, third-line treatment or fourth-line treatment. 8. The method according to any one of claims 1 to 3, wherein the compound of formula (I), gemcitabine and paclitaxel can be administered to the subject simultaneously, separately and/or sequentially. 9. Method according to any one of claims 1 to 7, wherein the compound of formula (I) is administered as a divided total dose. 10. Method according to any one of claims 1 to 7, wherein the compound of formula (I) is administered at a dose of about 240 mg twice daily.