WO2016161615A1

WO2016161615A1 - Method for treating cancer

Info

Publication number: WO2016161615A1
Application number: PCT/CN2015/076256
Authority: WO
Inventors: Kwan-Hwa Chi; Kai-lin YANG; Yu-Shan Wang; Hsin-Chien Chiang
Original assignee: Individual
Current assignee: Individual
Priority date: 2015-04-10
Filing date: 2015-04-10
Publication date: 2016-10-13
Anticipated expiration: 2017-10-10

Abstract

Disclosed herein is a method for treating a subject with a malignant solid tumor. The method includes administering to the subject at least one chemotherapeutic agent, and administering to the subject a chemotherapy sensitizer in an amount sufficient to enhance the subject responsiveness to the at least one chemotherapeutic agent. According to various embodiments of the present disclosure, the chemotherapy sensitizer consists of rapamycin and a substituted quinoline, such as hydroxychloroquine and chloroquine.

Description

METHDO FOR TREATING CANCER

BACKGROUND OF THE INVENTION

1. FIELD OF THE INVENTION

The present disclosure relates to field of cancer therapy. In particular, the present disclosure relates to the treatment for cancer patients who are resistant, non‐responsive, or refractory to previous cancer therapy.

2. DESCRIPTION OF RELATED ART

Cancer is a generic term for a large group of diseases characterized by the uncontrolled growth and spread of abnormal cells. These abnormal cells can then invade adjoining parts of the body and spread to other organs, which eventually may lead to death. Cancers is one of the leading causes of morbidity and mortality worldwide. In the United States, cancer is the second most common cause of death, exceeded only by heart disease, and accounts for nearly 1 of every 4 deaths. According to the estimation of the American Cancer Society, approximately 1.7 million new cancer cases are expected to be diagnosed in 2015, and each day, about 1, 620 Americans are expected to die of cancer.

Currently available treatments for cancer include surgery, radiation therapy, chemotherapy, hormone therapy, immunotherapy, targeted therapy, and photodynamic therapy, among others.

Chemotherapy (chemo) is the use of medicines or drugs to treat cancer. Unlike surgery and radiation, which are considered local treatments designated to remove or damage cancer cells in a certain area, chemotherapy is mostly used as a systemic treatment in which the chemotherapeutic agent kills not only the primary (original) tumor cells, but also the metastasized cancer cells that are spread to parts of the body away from the original tumor. Chemotherapy have been used to treat various cancers, and more than one hundred chemotherapeutic agents are used today, either alone or in combination with other chemotherapeutic agents or treatments. Despite the effort to develop more potent chemotherapy regimens with fewer side effects, currently available chemotherapy regimens for some types of cancers are associated with significant toxicity and minimal therapeutic gain.

In view of the foregoing, there exists a need in the art for providing a novel strategy for treating cancer patients who are not responsive to previous treatment (s) , such as hormone therapy, chemotherapy, combined chemotherapy, radio therapy, and/or targeted therapy.

SUMMARY

The following presents a simplified summary of the disclosure in order to provide a basic understanding to the reader. This summary is not an extensive overview of the disclosure and it does not identify key/critical elements of the present invention or delineate the scope of the present invention. Its sole purpose is to present some concepts disclosed herein in a simplified form as a prelude to the more detailed description that is presented later.

In one aspect, the present disclosure is directed to a method for treating a subject with a malignant solid tumor.

According to embodiments of the present disclosure, the method comprises the steps of administering to the subject at least one chemotherapeutic agent, and administering to the subject a chemotherapy sensitizer in an amount sufficient to enhance the subject responsiveness to the at least one chemotherapeutic agent. According to the present disclosure, the chemotherapy sensitizer consists of rapamycin and a substituted quinoline.

In certain embodiments, the substituted quinoline is hydroxychloroquine or chloroquine, or a pharmaceutically acceptable salt thereof. In one embodiment, the substituted quinoline is hydroxychloroquine.

According to some embodiments of the present disclosure, the rapamycin and the substituted quinolone are present in the chemotherapy sensitizer in a weight ratio between 10: 1 and 1: 5,000； preferably, 5: 1 to 1: 1,000； more preferably, 2: 1 to 1: 500 or 1: 1 to 1: 250.

In certain embodiments, the rapamycin is administered to the subject in an amount of 0.01 to 2.5 mg/kg body weight/day； preferably, 0.02 to 1 mg/kg body weight/day； more preferably, 0.04 to 0.5 mg/kg body weight/day.

According to various embodiments, the substituted quinoline is administered to the subject in an amount of 0.1 to 250 mg/kg body weight/day； preferably, 1 to 100 mg/kg body weight/day； more preferably, 5 to 50 mg/kg body weight/day.

In some embodiments, the rapamycin is administered to the subject in an amount of 0.04 to 0.5 mg/kg body weight/day, and the substituted quinoline is administered to the subject in an amount of 5 to 50 mg/kg body weight/day.

According to some embodiments of the present disclosure, each of the rapamycin and the substituted quinoline is administered once per day.

According to various embodiments of the present disclosure, the malignant solid tumor is any of the following, brain cancer, head and neck cancer, lung cancer, non‐small cell lung cancer, esophageal cancer, gastric cancer, hepatocellular carcinoma, small intestine cancer, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, renal cancer, bladder cancer, thyroid cancer, skin cancer, melanoma, and sarcoma.

In certain embodiments, the malignant solid tumor is a metastatic malignant solid tumor. Alternatively, or additionally, the malignant solid tumor is a refractory malignant solid tumor.

According to various embodiments of the present disclosure, the at least one chemotherapeutic agent is administered in a metronomic dosing.

In some embodiments, the subject is a human being.

Many of the attendant features and advantages of the present disclosure will becomes better understood with reference to the following detailed description considered in connection with the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

The present description will be better understood from the following detailed description read in light of the accompanying drawings, where:

Figure 1 provides images of the tumor of representative patients before and after metronomic chemotherapy, as well as after the salvage metronomic chemotherapy； and

Figure 2 provides images of the tumor of a representative patient after metronomic chemotherapy and the salvage metronomic chemotherapy.

DESCRIPTION

The detailed description provided below in connection with the appended drawings is intended as a description of the present examples and is not intended to represent the only forms in which the present example may be constructed or utilized. The description sets forth the functions of the example and the sequence of steps for constructing and operating the example. However, the same or equivalent functions and sequences may be accomplished by different examples.

For convenience, certain terms employed in the specification, examples and appended claims are collected here. Unless otherwise defined herein, scientific and technical terminologies employed in the present disclosure shall have the meanings that are commonly understood and used by one of ordinary skill in the art. Unless otherwise required by context, it will be understood that singular terms shall include plural forms of the same and plural terms shall include the singular. Specifically, as used herein and in the claims, the singular forms “a” and “an” include the plural reference unless the context clearly indicates otherwise. Also, as used herein and in the claims, the terms “at least one” and “one or more” have the same meaning and include one, two, three, or more.

Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in the respective testing measurements. Also, as used herein, the term “about” generally means within 10％, 5％, 1％, or 0.5％of a given value or range. Alternatively, the term “about” means within an acceptable standard error of the mean when considered by one of ordinary skill in the art. Other than in the operating/working examples, or unless otherwise expressly specified, all of the numerical ranges, amounts, values and percentages such as those for quantities of materials, durations of times, temperatures, operating conditions, ratios of amounts, and the likes thereof disclosed herein should be understood as modified in all instances by the term “about. ” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the present disclosure and attached claims are approximations that can vary as desired. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Ranges can be expressed herein as from one endpoint to another endpoint or between two endpoints. All ranges disclosed herein are inclusive of the endpoints, unless specified otherwise.

Throughout the present disclosure, the term "neoplasm" refers to a new and abnormal growth of cells or a growth of abnormal cells that reproduce faster than normal. A neoplasm creates an unstructured mass (a tumor) , which can be either benign or malignant. The term "benign" refers to a neoplasm or tumor that is noncancerous, e.g. its cells do not invade surrounding tissues or metastasize to distant sites； whereas the term "malignant" refers to a neoplasm or tumor that is metastatic, invades contiguous tissue or no longer under normal cellular growth control. Generally, the term "cancer" refers to all types of cancer or malignant neoplasm or tumors found in animals, including leukemia, carcinoma, melanoma, lymphomas, and sarcoma.

In the present disclosure, the term “solid tumors” refers a tumor which arises from or which develops in solid tissues that usually does not contain cysts or liquid areas. Examples of solid tumors are carcinomas, lymphomas, and sarcomas. Sarcomas are cancers arising from connective or supporting tissues such as bone or muscle； examples of sarcoma include, but are not limited to, fibrosarcoma, malignant fibrous hystiocytoma, liposarcoma, rhabdomyosarcoma, leiomyosarcoma, hemangiosarcoma, Kaposi's sarcoma, lymphangiosarcoma, synovial sarcoma, neurofibrosarcoma, extraskeletal chrondrosarcoma, extraskeletal osteosarcoma, embryonal sarcoma, alveolar sarcoma, dermatofibrosarcoma, infantile heamangiopericytoma, malignant peripheral nerve sheath tumors (also known as neurofibrosarcomas, malignant schwannomas, and neurogenic sarcomas) , alveolar soft part sarcoma, extraskeletal myxoid chondrosarcoma, and extraskeletal mesenchymal sarcoma. Carcinomas are cancers arising from glandular cells and epithelial cells, and illustrative examples of which include, colorectal carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatocellular carcinoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioblastoma multiforme, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, melanoma, neuroblastoma, and retinoblastoma. Lymphomas are cancers of the lymphoid organs such as the lymph nodes, spleen, and thymus； lymphomas are categorized as Hodgkin lymphomas or non‐Hodgkin lymphomas.

As used herein, the term "cancer therapy" refers to a therapy useful in treating cancer； in particular, malignant solid tumor. Examples of cancer therapy include, but are not limited to, e.g., surgery, radiation therapy, chemotherapy, hormone therapy, immunotherapy, targeted therapy, and photodynamic therapy.

The term "resistant" as used herein with reference to resistance to a cancer therapy (e.g., chemotherapy) refers to the fact that the normal therapeutic efficacy of the particular cancer therapy is not attained and that, for instance, a tumor continues to grow. For example, a subject resistant to chemotherapy is the one who suffers a relapse after at least one full cycle or course of cancer chemotherapy. In some cases, term "resistant to a treatment" means that the cell (e.g., tumor cell) or the subject exhibits reduced, diminished, or no responsiveness to a particular therapeutic agent (such as an EGFR‐Tyrosine kinase inhibitor) , as compared with the same cell or the subject at an earlier time point (in the case of acquired resistance or adaptive resistance) or as compared with other cells of the same type (known as natural resistance) or other subjects that respond to said therapeutic agent. According to certain embodiments of the present disclosure, a tumor is classified as “resistant” if the patient bearing the tumor was classified as “anon‐responder” according to, for instance, the criteria used or set forth by the European Organisation for Research and Treatment of Cancer (EORTC) in Response Evaluation Criteria in Solid Tumors (RECIST) , or evaluations using cancer‐specific antigen (e.g., postate‐specific antigen； PSA) or other tumor markers (such as, carcinoembryonic antigen (CEA) , and Serum cytokeratin fragment 21.1 (CFR 21.1) .

As used herein, the term "metastatic cancer" is to be understood as a disease in which at least one cancerous cell from a primary tumor has separated from the primary tumor and has continued to grow into a tumor at a location distinct from that of the primary tumor. The term "refractory cancers, " as used herein, refers to any type of cancer that either fails to respond favorably to a first line of cancer therapy, or alternatively, recurs or relapses after responding favorably to a first line of cancer therapy.

As used herein, the term “chemotherapy” refers to the administration of one or more chemotherapeutic agents to a patient in need thereof with the purpose to reduce, prevent, mitigate, limit, and/or delay the growth of neoplasms or metastases in the patient, or kill neoplastic cells directly by necrosis or apoptosis of neoplasms or any other mechanism. The term "chemotherapeutic agent, " as used herein refers to any chemical substance known to the clinical practitioner of ordinary skill in the art used for the treatment or amelioration of cancer and/or as an inducer of apoptosis in a patient.

Throughout the present disclosure and appended claims, the terms “treatment” and “treating” are used to include preventative (e.g., prophylactic) , curative, or palliative treatment that results in a desired pharmaceutical and/or physiological effect. Preferably, the effect is therapeutic in terms of partially or completely curing or preventing tumor genesis, progression, or metastasis. Also, the terms “treatment” and “treating” as used herein refer to the application or administration of the present pharmaceutical composition to a subject, who has been diagnosed with a malignant solid tumor or exhibits a symptom of or predisposition toward epidermoid carcinoma, with the purpose to partially or completely alleviate, ameliorate, relieve, delay onset of, inhibit progression of, reduce severity of, and/or reduce incidence of one or more symptoms or features of the malignant solid tumor. In particular, in the case where a patient is not responsive or resistant to a previous cancer therapy, the present treatment is useful in promoting the patient’s responsiveness to a chemotherapeutic treatment. Generally, a “treatment” includes not just the improvement of symptoms or decrease of markers of the disease, but also a cessation or slowing of progress or worsening of a symptom that would be expected in absence of treatment. Beneficial or desired clinical results of the present treatment include, but are not limited to, alleviation of one or more symptom (s) of the malignant solid tumor, diminishment of size and/or volume of the malignant solid tumor, stabilized (i.e., not worsening) state of the malignant solid tumor, or delay or slowing of progression, metastasis, or remission of the malignant solid tumor, whether partial or total, detectable or undetectable.

As used herein, the term "enhance the responsiveness" toward a chemotherapy is used in the present disclosure, and is meant to encompass the phenomenon in which when the tumor or the subject bearing the tumor has been determined to be not responsive to a previous cancer treatment, the same tumor or the subject bearing the tumor, after the present treatment, exhibits a measurable degree of responsiveness toward the chemotherapy.

The term “effective amount” as used herein refers to the quantity of a component (e.g., a chemotherapeutic agent) which is sufficient to yield a desired response (such as an inhibitory effect and/or a therapeutic effect) . As to the effective amount of a chemotherapy sensitizer, said effective amount is the quantity of the chemotherapy sensitizer that is sufficient to elicit a synergistic effect with a chemotherapeutic agent so that a subject (or tumor cell) that is not responsive to a previous cancer therapy exhibits a measurable degree of responsiveness toward the currant chemotherapy. Effective amount may be expressed, for example, in grams, milligrams, or micrograms, or as milligrams per kilogram of body weight (mg/kg) , milligrams per kilogram of body weight per day (mg/kg/day) or milligrams per day (mg/day) . The term also refers to an amount of a pharmaceutical composition containing an active component or combination of components. The specific effective or sufficient amount will vary with such factors as the particular condition being treated, the physical condition of the patient (e.g., the patient's body mass, age, or gender) , the type of mammal or animal being treated, the duration of the treatment, the nature of concurrent therapy (if any) , and the specific formulations employed and the structure of the compounds or its derivatives.

As used herein, the term "synergism" or "synergistic, " when referring to synergism between the chemotherapy sensitizer and a chemotherapy regimen, means that the two interact in ways that enhance or magnify one or more therapeutic and/or inhibitory effects of the chemotherapeutic agent (s) used in the chemotherapy regimen. That is, the combination of the two causes a greater effect than simply the sum of the individual effects of each component if they were used separately.

As used herein, a “pharmaceutically acceptable” component is one that is suitable for use with humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic response) commensurate with a reasonable benefit/risk ratio. Also, each excipient must be “acceptable” in the sense of being compatible with the other ingredients of the pharmaceutical formulation. The excipient can be in the form of a solid, semi‐solid, or liquid diluent, cream or a capsule.

The terms “subject” and “patient” are used interchangeably herein to refer to a mammal that is treatable with the present pharmaceutical composition and/or method. The term “mammal” refers to all members of the class Mammalia, including humans, primates, domestic and farm animals (such as rabbit, pig, sheep, and cattle) , rodents (such as mouse, rat, guinea pig, marmots, hamster) , and zoo, sports or pet animals. The term “subject” is intended to refer to both the male and female gender unless one gender is specifically indicated. In some embodiments, the subject is a human being with a malignant solid tumor that is resistant or non‐responsive to a previous cancer therapy.

As used herein, the term "clinical benefit" in the context of treating human solid tumor refers to a statistically significant decrease in at least one of: the rate of tumor growth, a cessation of tumor growth, or in a reduction in the size, mass, metabolic activity, or volume of the tumor, as measured by standard criteria described below.

The present disclosure is based, at least, on the finding that the co‐administration of the present chemotherapy sensitizer and one or more chemotherapeutic agent (s) synergistically increase the antitumor efficacy of the chemotherapeutic agent (s) in cases where the patients failed to respond to a previous cancer therapy. As could be appreciated by persons having ordinary skill in the art, since these patients were not responsive to the previously administered cancer therapy, the subsequent chemotherapy is not expected to produce any positive outcome in most cases. However, the clinical trial (as provided below) conducted on patients having malignant solid tumors and unresponsive to previous cancer treatment (s) , indicates that the co‐administration of the chemotherapy sensitizer and chemotherapeutic agent (s) may enhance the patients’ responsiveness to the chemotherapeutic agent (s) in all cases. This synergistic effect achieved by the chemotherapy sensitizer and the chemotherapeutic agent (s) is unexpected to persons having ordinary skill in the art. Accordingly, the first aspect of the present disclosure is directed to a method for treating a subject with a malignant solid tumor using the present chemotherapy sensitizer in combination with a chemotherapy regimen.

According to embodiments of the present disclosure, the method comprises the steps of administering to the subject at least one chemotherapeutic agent, and administering to the subject a chemotherapy sensitizer in an amount sufficient to enhance the subject responsiveness to the chemotherapeutic treatment. Specifically, the chemotherapy sensitizer consists of rapamycin and a substituted quinoline.

The substituted quinolines useful in the present invention may be prepared by synthetic techniques that are well known in the art and there have been many commercially‐available substituted quinolines. In certain embodiments, suitable substituted quinolines include 4‐aminoquinoline, 8‐aminoquinoline, and hydroxymethylquinoline, hydroxychloroquine, chloroquine, amodiaquine, amopyroquine, bis‐pyroquine, cycloquine, oxychloroquine, sontoquine, amoproquine, primaquine, mefloquine, quinacrine, tebuquine, quinine, thalidomide, sulfasalazine, and sulfapyridine, or pharmaceutically acceptable salt or enantiomer or prodrug thereof. A particularly example of preferred substituted quinolines used in the present examples is hydroxychloroquine or a pharmaceutically acceptable salt thereof.

According to some embodiments of the present disclosure, the rapamycin and the substituted quinoline are present in the chemotherapy sensitizer in a weight ratio of about 10: 1 to 1: 5,000； preferably, 5: 1 to 1: 1,000； more preferably, 2: 1 to 1: 500 or 1: 1 to 1: 250. For example, the weight ratio between the rapamycin and the substituted quinoline is 10, 9.5, 9, 8.5, 8, 7.5, 7, 6.5, 6, 5.5, 5, 4.5, 4, 3.5, 3, 2.5, 2, 1.9, 1.8, 1.7, 1.6, 1.5, 1.4, 1.3, or 1.2 : 1, or 1: 1, 1.25, 1.75, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700, 1,800, 1,900, 2,000, 2,500, 3,000, 3,500, 4,000, 4,500, or 5,000.

According to some embodiments of the present disclosure, the rapamycin is administered to the subject in an amount of 0.01 to 2.5 mg/kg body weight/day； preferably, 0.02 to 1 mg/kg body weight/day； more preferably, 0.04 to 0.5 mg/kg body weight/day. For example, the rapamycin is administered to the subject in an amount of 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, or 2.5 mg/kg body weight/day. Alternatively, the rapamycin is administered to the subject in an amount of 1 to 100 mg per day.

According to various embodiments, the substituted quinoline is administered to the subject in an amount of 0.1 to 250 mg/kg body weight/day； preferably, 1 to 100 mg/kg body weight/day； more preferably, 5 to 50 mg/kg body weight/day. Specifically, the substituted quinoline is administered to the subject in an amount of 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 200, or 250 mg/kg body weight/day. Alternatively, the substituted quinoline is administered to the subject in an amount of 10 mg to 10 g per day.

In some embodiments, the rapamycin is administered to the subject in an amount of 0.04 to 0.5 mg/kg body weight/day, and the substituted quinoline is administered to the subject in an amount of 5 to 50 mg/kg body weight/day. Alternatively, the rapamycin is administered to the subject in an amount of 1.5 to 50 mg/day, and the substituted quinoline is administered to the subject in an amount of 200 mg/day to 5 g/day.

According to some embodiments of the present disclosure, each of the rapamycin and the substituted quinoline is administered once per day. However, the present disclosure is not limited thereto, and any suitable administration interval may be adopted by the medical practitioner performing the present method. Further, the rapamycin and the substituted quinoline may be administered at the same or different intervals depending on the actual need.

Chemotherapeutic agent (s) suitable to be used in the present method include, but are not limited to, alkylating agents, platinum drugs, antimetabolites, anti‐tumor antibiotics, topoisomerase inhibitors, mitotic inhibitors, corticosteroids, and other miscellaneous chemotherapeutic agents. Alkylating agents directly damage DNA to prevent the cancer cell from reproducing； illustrative examples of which include, nitrogen mustards (such as, mechlorethamine (nitrogen mustard) , chlorambucil, cyclophosphamide

ifosfamide, and melphalan) , nitrosoureas (including streptozocin, carmustine (BCNU) , and lomustine) , alkyl sulfonates (e.g., busulfan) , triazines (such as, dacarbazine (DTIC) and temozolomide

) , and ethylenimines (e.g., thiotepa and altretamine (hexamethylmelamine) ) . The platinum drugs are sometimes grouped with alkylating agents because they kill cells in a similar way； examples of platinum drugs include cisplatin, carboplatin, and oxalaplatin. Antimetabolites interfere with DNA and RNA growth by substituting for the normal building blocks of RNA and DNA； examples of antimetabolites include, 5‐fluorouracil (5‐FU) , 6‐mercaptopurine (6‐MP) , capecitabine

cladribine, clofarabine, cytarabine

foxuridine, fludarabine, gemcitabine

hydroxyurea, methotrexate, pemetrexed

pentostatin, and thioguanine. Anti‐tumor antibiotics either break down DNA strands or slow down or stop DNA synthesis, thereby retarding the proliferation of cancer cells； currently available anti‐tumor antibiotics include anthracyclines (such as, daunorubicin, doxorubicin

epirubicin, and idarubicin) , actinomycin‐D, bleomycin, dactinomycin, mitomycin‐C, and plicamycin. Topoisomerase inhibitors interfere with topoisomerases, which help separate the strands of DNA during cell proliferation； examples of topoisomerase I inhibitors include camptothecin (CPT) , irinotecan (CPT‐11) , and topotecan； examples of topoisomerase II inhibitors include etoposide (VP‐16) , mitoxantrone, and teniposide. Mitotic inhibitors can stop mitosis or inhibit the synthesis of proteins involved in cell reproduction； examples of mitotic inhibitors include, taxanes (such as, paclitaxel

and docetaxel

) , epothilones, (e.g., ixabepilone

) , vinca alkaloids (such as, vinblastine

vincristine

and vinorelbine

) , and estramustine

Corticosteroid chemotherapeutic agents include natural hormones and hormone‐like drugs capable of killing cancer cells or slowing their growth, examples include prednisone, methylprednisolone

and dexamethasone

Other miscellaneous chemotherapy drugs that act in slightly different ways and do not fit well into any of the other categories include, but are not limited to, L‐asparaginase and the proteosome inhibitor bortezomib

As could be appreciated, some chemotherapeutic agents may fall into more than two categories described above depending on the source or chemical structure, or the way in which they act. For example, mitoxantrone can be considered an anti‐tumor antibiotic, which also acts as a topoisomerase II inhibitor. Accordingly, the above‐provided classification should not be regarded as a limitation to the claimed invention. Further, persons having ordinary skill in the art will appreciate that the above list is not exhaustive, and other chemotherapeutic agents (currently available ones or those might be developed in the future) are also envisaged in the scope of the present disclosure, as long as the therapeutic efficacy of these chemotherapeutic agents is boosted by the present chemotherapy sensitizer.

While administering the chemotherapy sensitizer or the at least one chemotherapeutic agent (s) , the chemotherapy sensitizer or the at least one chemotherapeutic agent (s) is preferably formulated into one or more pharmaceutical compositions suitable for administration. According to various embodiments of the present disclosure, the chemotherapy sensitizer and the at least one chemotherapeutic agent can be formulated in a single pharmaceutical composition or in separate pharmaceutical compositions. Still optionally, the rapamycin and the substituted quinoline can be formulated in a single pharmaceutical composition or in separate pharmaceutical compositions. The above‐mentioned preparation (s) can be prepared in accordance with acceptable pharmaceutical procedures, such as described in Remington: The Science and Practice of Pharmacy, 20th edition, ed. Alfonoso R. Gennaro, Lippincott Williams &Wilkins (2000) .

Each pharmaceutical composition may comprise at least one pharmaceutically acceptable excipient that is compatible with other ingredient (s) in the formulation and biologically acceptable. The choice of a pharmaceutically acceptable excipient to be used in conjunction with the components of the present composition is basically determined by the way the pharmaceutical composition is to be administered.

The chemotherapy sensitizer, the chemotherapeutic agent, or the pharmaceutical composition (s) comprising at least one of the above‐mentioned components may be administered by any suitable route, for example, by oral or parenteral (such as, intravenous, subcutaneous, intramuscular, intrathecal, intraperitoneal, intrarectal, viginal, nasal, intragastric, intratracheal, pulmonary, intratumoral or peritumoral injection) administration. Alternatively, the pharmaceutical composition or components thereof may be administered via an implant. As could be appreciated, in the case where the three components (i.e., rapamycin, substituted quinoline, and at least one chemotherapeutic agent) are provided in at least two dose units, these dose units may be administered via the same or different routes. Still optionally, these dose units may be given simultaneously or sequentially in time. Also, each of these components may be given with equal or different time intervals.

For example, in the case where the chemotherapy sensitizer is formulated in one dose unit, while the chemotherapeutic agent is formulated in another dose unit, the chemotherapy sensitizer may be administered prior to, concurrently with, or after the administration of the chemotherapeutic agent. When the two dose units are not administered concurrently, the time interval between the administrations of said two dose units is 12 hours at most. For example, the time interval nay be 1 2, 3, 4, 5, 10, 15, 20, or 25 minutes, or 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, or 12 hours. Preferably, the time interval may be less than 6 hours； more preferably, less than 5 hours.

Further, the chemotherapy sensitizer and chemotherapeutic agent can be administered at the same or different frequencies. For example, during one course of the chemotherapy, the chemotherapy sensitizer may be given on a daily basis while the chemotherapeutic agent is administered on an intermittent basis (e.g., every other day) . As could be appreciated by persons having ordinary skill in the art, the present invention is not limited to the above‐mentioned dosage regimens, which are provided for illustrative purpose only.

The above‐mentioned dosage regimes are also applicable in case where the three components are formulated in three different dosage units. For example, the dosage units of substituted quinoline and rapamycin are administered at substantially the same time and the same frequency, while the dosage unit of chemotherapeutic agent is administered at the same time or different time, with the same or a different frequency.

According to embodiments of the present disclosure, the chemotherapy protocols suitable for use herein include, but are not limited to, adjuvant chemotherapy, neoadjuvant chemotherapy induction chemotherapy, consolidation chemotherapy, and maintenance chemotherapy. Adjuvant chemotherapy is given to destroy left‐over (microscopic) cells that may be present after the known tumor is removed by surgery so as to prevent a possible cancer reoccurrence. Neoadjuvant chemotherapy is performed prior to the surgical procedure mostly in an attempt to shrink the tumor so that the surgical procedure may not need to be as extensive. Induction chemotherapy is administered to induce a remission, while consolidation chemotherapy (or intensification therapy) is performed once a remission is achieved to sustain the remission. Maintenance chemotherapy is given in lower doses to assist in prolonging a remission.

According to various embodiments of the present disclosure, the at least one chemotherapeutic agent is administered to the subject at a metronomic dosing. As used here, the term "metronomic dosing" refers to a long‐term, low‐dose, frequent administration of the chemotherapeutic drug (s) . For example, the chemotherapeutic drug (s) may be administered using a dose lower than the maximum tolerated doses (MTD) of a chemotherapeutic. Specifically, the metronomic dose may be one‐tenth to one‐third of the MTD.

In some embodiments, the subject is a human being.

The following Examples are provided to elucidate certain aspects of the present invention and to aid those of skilled in the art in practicing this invention. These Examples are in no way to be considered to limit the scope of the invention in any manner. Without further elaboration, it is believed that one skilled in the art can, based on the description herein, utilize the present invention to its fullest extent. All publications cited herein are hereby incorporated by reference in their entirety.

1. Patient Recruitment

From May 2012 to September 2014, a total of 46 patient volunteers were recruited at Shin Kong Wu Ho‐Su Memorial Hospital, Taipei, Taiwan, under the approval of the Institutional Review Board of said Hospital with written informed consent of the patients. To be included in the following analysis, each recruited patients shall have incurable metastatic/recurrent disease and does not respond to current metronomic chemotherapy regardless of the primary tumor type he/she has. Specifically, the enrollment criteria included: Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1； life expectancy of at least 3 months before the salvage treatment； at least one single site of measurable (two‐dimensional) disease or serial and continuous (>3 monthly measurement) elevated tumor markers (at least twice the upper limit) in patients with non‐measurable lesions. Patients were excluded if treatment with the metronomic therapy and salvage therapy was shorter than 12 weeks, unless there was a radiographic confirmation of disease progression. Twenty‐five (17 female, 8 male) of the enrolled patients satisfying the above‐mentioned criteria were identified and clinical data of these patients were used for further analysis. Demographics of the enrolled patients are summarized in Table 1, below.

Table 1

2. Patient Treatment and Assessment

Before the enrollment, patients had been treated with the metronomic regimen suitable for their respective tumor types (see, Table 2, below) . The same metronomic regimen for each patient was continued after the enrollment, in conjunction with 2 mg rapamycin (oral, once daily) and 400 mg hydroxychloroquine (oral, once daily) as the salvage treatment. Treatment was discontinued with the development of grade IV non‐myelotoxicity, patient intolerance, or disease progression.

Treatment‐related toxicity was assessed every two weeks. Toxicity was scored according to the Common Terminology Criteria for Adverse Events (CTCAE) v4.0.

Patients were examined using chest radiography, computed tomography (CT) , or positron emission tomography–computed tomography, every 2 to 3 months from the time of enrollment. Tumor markers were assessed every 1 to 3 months.

Response and progression were evaluated using the international criteria proposed by the Response Evaluation Criteria in Solid Tumors (RECIST) guideline (version 1.1) based on the examining results determined in accordance with the above‐mentioned procedures. The response was categorized into one of the following RECIST criteria, complete response (CR) , partial response (PR) , progressive disease (PD) and stable disease (SD) . In patients with complete response (CR) , no measurable disease was observed and there were no development of new lesions, with tumor markers dropping to the normal range. As to patients with partial response (PR) , at least 30％reduction in the sum of the longest diameters measured at disease sites or enhanced area was recorded. Patients categorized into the progressive disease (PD) group experienced at least 20％increase in the sum of the longest diameter measured disease sites or appearance of new lesions. For patients not falling into any of the criteria of CR, PR, or PD, or for patients with tumor markers decline of > 50％in non‐measurable lesions and stable disease, these patients were determined to have stable disease (SD) .

Clinical benefit rate was defined as the percentage of patients without disease progression for more than 3 months.

3. Results

Among the 25 patients underwent evaluation, 10 patients (40％) experienced PR, 11 patients (44％) had SD, and only 4 patients exhibited disease progression (patient #7, 21, 22, and 25) . That is, eighty‐four percent of patients experienced clinical benefits (SD+PR) for more than 3 months. The clinical characteristics and results of patients who received this treatment strategy are summarized in Table 2. The median follow‐up time was 11 months (ranging from 3 to 28 months) . The median duration of salvage treatment was 4 months (95％confidence interval, 3–7 months) before disease progression, contented stop, or refusal to continue treatment.

It was difficult to evaluate the effect of the addition of chemotherapy sensitizer (rapamycin and hydroxychloroquine) to metronomic chemotherapy on the progression‐free survival (PFS) in such a heterogeneous group of patients. Nevertheless, the results in Table 2 indicated that 2 patients progressed from PD to PR and another 8 patients progressed from SD to PR after the salvage treatment, suggesting this treatment being a potential alternative treatment solution for patients not responding to conventional chemotherapy.

Images of the tumor of representative patients before and after metronomic chemotherapy, as well as after the salvage metronomic chemotherapy are provided in Figure 1 (

patient #

6, 8, 9, 19, and 20) and Figure 2 (patient #13) .

Patient #6 had bladder cancer with multiple liver metastases, and referring to Figure 1, although the tumor did not reduce in size, the sum of diameters of enhancement regions were reduced by more than 30％, according to the modified RECIST criteria. Patient #8 had a nodular lesion in right upper lung (black arrow in Figure 1) , the size of which was significantly reduced after the salvage treatment. Patient #9 had right pleural seeding tumor (white arrow in Figure 1) , the size of which was also significantly reduced after the salvage treatment. Both patient #19 and patient #20 experienced bilateral multiple lung metastases, and the metastases were reduced after the salvage treatment.

Table 2

* Previous treatment after initial treatment failure.

Abbreviations: Ca, cancer； meta, metastasis； bid, twice a day； tid, three times a day； qd, every day； qod, every weeks； LN, lymph node(s)； CCRT, concomitant chemoradiotherapy； TKI, tyrosine kinase inhibitor； RT, radiother fluorouracil and folinic acid； FOLFOX, oxaliplatin with fluorouracil and folinic acid； CAF, 5‐fluorouracil, doxorubi

Patient #13 had peritoneal seeding in the right pelvic region, and the positron emission tomography–computed tomography scanning image of Figure 2 indicated a significant shrinkage of the tumor after the salvage treatment.

Data related to non‐hematologic toxicity of the present salvage treatment were summarized in Table 3, below.

Table 3

The data above indicated that the present salvage treatment was well tolerated. In particular, 8 patients (32％) reported grade ≥1 fatigue, including 1 patient who had grade 3 fatigue and had to discontinue the treatment. Diarrhea was the second most common side effects of the chemotherapy, with 4 (16％) patients exhibiting grade 2 diarrhea, and 1 (4％) patient reporting grade 3 diarrhea. Two (8％) patients had mucositis, and 1 (4％) reported grade 3 nausea/vomiting or renal toxicity. Myelotoxicity was relatively common, with 8 patients (32％) developing grade ≥ 3 thrombocytopenia, 6 patients (24％) developing grade ≥ 3 leucopenia and 3 patients (12％) having grade ≥ 3 anemia.

One patient (patient #23) had grade 3 cardiotoxicity, with a left ventricle ejection fraction of 35％； this patient had no history of doxorubicin usage, and recovered after discontinuing all treatment.

Another patient (patient #22) experienced grade V hepatitis, which was attributed to the reactivation of previously unnoted hepatitis B virus. Patient #22 had not been administered prophylactic anti‐viral medicine.

None of these patients developed febrile neutropenia, and they all recovered quickly from myelotoxicities after one to two weeks of treatment interruption.

The above‐mentioned clinical data established that the addition of rapamycin and hydroxychloroquine to conventional metronomic chemotherapy was well tolerated in patients respectively having various types of cancers and was safe to the patient. Most importantly, by co‐administrating the chemotherapy sensitizer and one or more chemotherapeutic agents, response rate to treatment increased by 40％, and the cancers became stabilized in 84％patients in a cohort of patients refractory to their chemotherapy regimen. This significant clinical benefit observed in patients resistant to chemotherapy, was unexpected.

The results of this self‐controlled study indicated that co‐administration of a chemotherapy sensitizer and one or more chemotherapeutic agent was not only effective in preventing tumors from progressing, in some cases, it could also reverse therapeutic drug resistance. The collective data as presented herein suggests that the autophagy inducer (e.g., rapamycin) and lysosomal inhibitor (e.g., hydroxychloroquine) act synergistically withthe conventional chemotherapeutic agent (s) .

In sum, the clinical data provided hereinabove established that the co‐administration of a chemotherapy sensitizer (which consists of rapamycin and a substituted quinoline, such as hydroxychloroquine) and a chemotherapy regimen (e.g., metronomic chemotherapy) to a subject who is resistant, non‐responsive or refractory to a previous cancer therapy, will enhance the subject’s responsiveness to the at least one chemotherapy regimen. In other words, the responsiveness of the resistant, non‐responsive or refractory tumor of a subject is improved in a measurable degree under the sensitizing action of rapamycin and the substituted chloroquine (e.g., hydroxychloroquine) .

It will be understood that the above description of embodiments is given by way of example only and that various modifications may be made by those with ordinary skill in the art. The above specification, examples, and data provide a complete description of the structure and use of exemplary embodiments of the invention. Although various embodiments of the invention have been described above with a certain degree of particularity, or with reference to one or more individual embodiments, those with ordinary skill in the art could make numerous alterations to the disclosed embodiments without departing from the spirit or scope of this invention.

Claims

A method for treating a subject with a malignant solid tumor, comprising,

administering to the subject at least one chemotherapeutic agent； and

administering to the subject a chemotherapy sensitizer consisting of rapamycin and a substituted quinoline in an amount sufficient to enhance the responsiveness of the malignant solid tumor to the at least one chemotherapeutic agent.
The method of claim 1, wherein the substituted quinoline is hydroxychloroquine or chloroquine, or a pharmaceutically acceptable salt thereof.
The method of claim 2, wherein the substituted quinoline is hydroxychloroquine.
The method of claim 1, wherein the rapamycin and the substituted quinoline are present in the chemotherapy sensitizer in a weight ratio of 10: 1 to 1: 5, 000.
The method of claim 4, wherein the rapamycin and the substituted quinoline are present in the chemotherapy sensitizer in a weight ratio of 5: 1 to 1: 1, 000.
The method of claim 5, wherein the rapamycin and the substituted quinoline are present in the chemotherapy sensitizer in a weight ratio of 2: 1 to 1: 500.
The method of claim 6, wherein the rapamycin and the substituted quinoline are present in the chemotherapy sensitizer in a weight ratio of 1: 1 to 1: 250.
The method of claim 1, wherein the rapamycin is administered to the subject in an amount of 0.01 to 2.5 mg/kg body weight/day.
The method of claim 1, wherein the rapamycin is administered to the subject in an amount of 0.02 to 1 mg/kg body weight/day.
The method of claim 1, wherein the rapamycin is administered to the subject in an amount of 0.04 to 0.5 mg/kg body weight/day.
The method of claim 1, wherein the substituted quinoline is administered to the subject in an amount of 0.1 to 250 mg/kg body weight/day.
The method of claim 1, wherein the substituted quinoline is administered to the subject in an amount of 1 to 100 mg/kg body weight/day.
The method of claim 1, wherein the substituted quinoline is administered to the subject in an amount of 5 to 50 mg/kg body weight/day.
The method of claim 1, wherein the rapamycin is administered to the subject in an amount of 0.04 to 0.5 mg/kg body weight/day, and the substituted quinoline is administered to the subject in an amount of 5 to 50 mg/kg body weight/day.
The method of claim 14, wherein the rapamycin and the substituted quinoline are administered once per day, respectively.
The method of claim 1, wherein the malignant solid tumor is brain cancer, head and neck cancer, lung cancer, non‐small cell lung cancer, esophageal cancer, gastric cancer, hepatocellular carcinoma, small intestine cancer, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, renal cancer, bladder cancer, thyroid cancer, skin cancer, melanoma, or sarcoma.
The method of claim 1, wherein the malignant solid tumor is a metastatic malignant solid tumor.
The method of claim 1, wherein the malignant solid tumor is a refractory malignant solid tumor.
The method of claim 1, wherein the at least one chemotherapeutic agent is administered in a metronomic manner.
The method of claim 1, wherein the subject is a human being.