Hyaluronan-Nimesulide Conjugates for Treating Locally Advanced or Metastatic Tumors - Patent application

Abstract

Compositions and methods for the treatment of locally advanced or metastatic tumors are disclosed. According to some embodiments, the compositions and methods include the use of hyaluronan-nimesulide conjugates in combination with pyrimidine-based nucleotide analogs.

Description

[Technical field]

[0001] The present disclosure relates to compositions and methods for treating locally advanced or metastatic solid or liquid tumors in patients. More particularly, the compositions and methods include the use of hyaluronan-nimesulide conjugates in the treatment of locally advanced or metastatic solid or liquid tumors. [Background technology]

[0002] Nimesulide is an oral nonsteroidal anti-inflammatory drug (NSAID) and selective cyclooxygenase-2 (COX-2) inhibitor that has been used since 1985 as an analgesic and antipyretic for painful musculoskeletal disorders (osteoarthritis) and primary dysmenorrhea. Since the late 1990s, numerous reports of nimesulide-induced liver injury have appeared in the medical literature. These reports have led to the withdrawal of the drug's approval in some countries and its removal from approval in others. After a comprehensive evaluation of the drug's safety profile by the European Medicines Agency, the Committee for Medicinal Products for Human Use recommended the restriction of indications for nimesulide, withdrawal of the 200 mg formulation from the market, and restriction to a dose of 100 mg twice daily. Symptoms of nimesulide-induced liver injury include jaundice, fatigue, and pruritus, and are generally associated with prolonged use beyond 15 days. More recently, preclinical studies have demonstrated that nimesulide may have anticancer activity in many types of tumors, but its poor solubility limits its efficacy and its potential as a primary anticancer drug.

[0003] Meanwhile, cancer is the leading cause of death worldwide, with approximately 10 million cancer deaths in 2020. Patients with locally advanced or metastatic tumors often have little chance of recovery. Summary of the Invention [Problem to be solved by the invention]

[0004] In view of the above, there exists a need in the art to provide additional effective compositions and methods for treating locally advanced or metastatic solid or liquid tumors. [Means for solving the problem]

[0005] (overview) The following presents a simplified summary of the disclosure in order to provide the reader with a basic understanding. This summary is not an extensive overview of the disclosure, and it does not identify key / critical elements of the invention or delineate the scope of the 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 below.

[0006] In one embodiment, the disclosure relates to a method for treating a locally advanced or metastatic tumor in a patient in need of treatment.

[0007] According to some embodiments of the present disclosure, the method includes administering to a patient an effective amount of a hyaluronan-nimesulide conjugate in combination with an effective amount of a pyrimidine-based nucleotide analog, such as a fluoropyrimidine.

[0008] According to some embodiments of the present disclosure, the locally advanced or metastatic tumor may be a tumor including a locally advanced or metastatic sarcoma, pancreatic tumor, colorectal tumor, liver tumor, melanoma, lung tumor, breast tumor, ovarian tumor, head or neck tumor, gastric tumor, prostate tumor, esophageal tumor, cervical tumor or vaginal tumor, brain tumor (such as glioblastoma, astrocytoma, meningioma or medulloblastoma). Alternatively, the locally advanced or metastatic tumor may be a liquid tumor including a locally advanced or metastatic multiple myeloma, leukemia, or lymphoma.

[0009] In certain embodiments, the hyaluronan-nimesulide conjugate is administered intravenously to a patient at 0.1-3 mg / kg Nimesulide equivalent (Nim equivalent) per administration. In certain embodiments, the hyaluronan-nimesulide conjugate is administered intravenously to a patient at 5-150 mg / kg hyaluronan-nimesulide conjugate per administration. In certain embodiments, the hyaluronan-nimesulide conjugate is administered at predetermined intervals during the treatment period. In certain embodiments, the hyaluronan-nimesulide conjugate is administered twice (on days 1 and 15) during a 28-day cycle.

[0010] According to certain embodiments of the present disclosure, the fluoropyrimidine is 5-fluorouracil (5-FU), 5-fluorocytosine, 5-fluorothymidine, capecitabine, and trifluorothymidine, carmofur, doxifluridine, emitefur, tegafur, or floxuridine. In certain embodiments, the fluoropyrimidine is 5-FU. In certain embodiments, the 5-FU is administered to the patient at 1-30 mg / kg / dose. In certain embodiments, the 5-FU is administered to the patient intravenously once a week.

[0011] In another aspect, the present disclosure relates to a pharmaceutical composition for treating locally advanced or metastatic tumors.

[0012] According to some embodiments, the pharmaceutical composition comprises an effective amount of a hyaluronan-nimesulide conjugate and a pharma- ceutically acceptable excipient, wherein the pharmaceutical composition is used in combination with a pyrimidine-based nucleotide analog, such as a fluoropyrimidine.

[0013] Subject matter encompassed within other aspects of the present disclosure includes the use of a hyaluronan-nimesulide conjugate in the manufacture of a medicament for use in the treatment of locally advanced or metastatic tumors, and a hyaluronan-nimesulide conjugate for use in the treatment of locally advanced or metastatic tumors, wherein the medicament or the hyaluronan-nimesulide conjugate is used in combination with a pyrimidine-based nucleotide analogue, such as a fluoropyrimidine.

[0014] Many of the attendant features and advantages of the present disclosure will be better understood by reference to the following detailed description considered in conjunction with the accompanying drawings.

[0015] The specification will be better understood from the following detailed description taken in light of the accompanying drawings. [Brief description of the drawings]

[0016] [Figure 1A] 1 is a line graph showing the effect of combined administration of CA102N and 5-FU on cell proliferation of HT-29 cells according to one embodiment of the present disclosure. [Figure 1B] 1 is a line graph showing the effect of combined administration of CA102N and 5-FU on cell proliferation of HT-29 cells according to one embodiment of the present disclosure. [Diagram 2] 1 is a line graph showing the effect of combined administration of CA102N and 5-FU on tumor volume in CT26 mice, according to one embodiment of the present disclosure. [Diagram 3] 1 is a line graph showing the effect of combined administration of CA102N and 5-FU on tumor volume in CR5038PDX mice, according to one embodiment of the present disclosure. [Figure 4] 1 is a line graph showing the effect of combined administration of CA102N and 5-FU on tumor volume in HT-29 tumor-bearing mice, according to one embodiment of the present disclosure. [Diagram 5]1 is a line graph showing the effect of combined administration of CA102N and 5-FU on tumor volume in MDA-MB-231 tumor-bearing mice, according to one embodiment of the present disclosure. [Figure 6] 1 is a bar graph showing the effect of combined administration of CA102N and 5-FU on cell proliferation of Bx-PC-3 cells, according to one embodiment of the present disclosure. [Figure 7] 1 is a bar graph showing the effect of combined administration of CA102N and 5-FU on cell proliferation of PAC-1 cells, according to one embodiment of the present disclosure. [Figure 8] 1 is a bar graph showing the effect of combined administration of CA102N and 5-FU on cell proliferation of A549 cells, according to one embodiment of the present disclosure. [Figure 9] 1 is a bar graph showing the effect of combined administration of CA102N and 5-FU on cell proliferation of MM.1S cells, according to one embodiment of the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] (explanation) The detailed description provided below in conjunction with the accompanying drawings is intended to be illustrative of the present embodiment and is not intended to represent the only manner in which the present embodiment may be constructed or utilized. The description sets forth the functions of the present embodiment and the sequence of steps for constructing and executing the present embodiment. However, the same or equivalent functions and sequences may be accomplished by different embodiments.

[0018] For convenience, certain terms employed in the specification, examples, and appended claims are collected here. Unless otherwise defined herein, scientific and technical terms employed in this disclosure shall have the meanings commonly understood and used by those skilled in the art. It is understood that the singular includes the plural of the same and the plural includes the singular, unless the context otherwise requires. Specifically, as used in the specification and claims, the singular forms "a" and "an" also include the plural, unless the context clearly indicates otherwise. Also, as used in the specification and claims, the terms "at least one" and "one or more" have the same meaning and include one, two, three, or more.

[0019] 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. However, any numerical value contains certain errors resulting from the standard deviation found in their respective testing measurements. Additionally, as used herein, the term "about" generally means within 10%, 5%, 1%, or 0.5% of a given value or range. Additionally, the term "about" means within an acceptable standard error of the mean as would be expected by one of ordinary skill in the art. Unless otherwise specified in the runs / examples or otherwise expressly indicated, all numerical ranges, amounts, values, and percentages for amounts of materials, lengths of time, temperatures, run conditions, proportions of amounts, and the like disclosed herein are to be understood as being modified in all instances by the term "about." Thus, unless specifically indicated to the contrary, the numerical parameters set forth in the present disclosure and appended claims are approximations that may be varied as desired. At a minimum, each numerical parameter should be interpreted at least in light of the number of reported significant digits and by applying ordinary rounding techniques. Ranges may be expressed herein as from one endpoint to another endpoint or between two endpoints. All ranges disclosed herein include the endpoints unless otherwise specified.

[0020] The terms "treatment" and "treating" as used herein may refer to preventative (e.g., prophylactic), curative or palliative measures. In particular, the term "treating" as used herein refers to the application or administration of the hyaluronan-nimesulide conjugate or a pharmaceutical composition comprising the same to a subject with a pathology, a symptom associated with a pathology, a disease or disorder secondary to a pathology, or a predisposition to a pathology, with the purpose of partially or completely alleviating, improving, relieving, delaying the onset, inhibiting the progression, reducing the severity, and / or reducing the incidence of one or more symptoms or characteristics of the particular disease, disorder, and / or condition. Treatment may be administered to subjects who do not show signs of a disease, disorder, and / or condition and / or to subjects who show only early signs of a disease, disorder, and / or condition, with the purpose of reducing the risk of developing pathologies associated with the disease, disorder, and / or condition.

[0021] The terms "subject" and "patient" are used interchangeably herein and are intended to refer to animals, including the human species, treatable by the hyaluronan-nimesulide conjugates, pharmaceutical compositions, and / or methods of the present invention. The term "subject" or "patient" is intended to refer to both male and female genders, unless one gender is specifically indicated.

[0022] The terms "application" and "administration" are used interchangeably herein and refer to the application of a hyaluronan-nimesulide conjugate or pharmaceutical composition of the present invention to a subject in need of treatment.

[0023] The term "effective amount" as used herein refers to an amount of the hyaluronan-nimesulide conjugate of the present invention sufficient to produce a desired therapeutic effect. An effective amount of a drug is not required to cure a disease or condition, but provides treatment for a disease or condition, such as delaying, hindering, or preventing the onset of the disease or condition, or ameliorating the symptoms of the disease or condition. The effective amount may be divided into one, two, or more doses in a form suitable for administration once, twice, or more times over a specified period of time. The specific effective or sufficient amount will vary depending on factors such as the particular condition being treated, the physical condition of the patient (e.g., the patient's weight, age, or sex), the type of mammal or animal being treated, the duration of treatment, the characteristics of any concomitant therapy, and the specific dosage form and structure of the compound or its derivatives used. The effective amount may be expressed, for example, as the total mass of the ester prodrug (e.g., grams, milligrams, or micrograms) or as the ratio of the mass of the ester prodrug to body weight (e.g., milligrams per kilogram (mg / kg)).

[0024] The term "pharmaceutical acceptable excipient" as used herein means a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, carrier, solvent, encapsulating material, etc., involved in carrying or transporting a subject agent, such as from one organ or part of the body to another. Each excipient must be "acceptable" in the sense of being compatible with the other components of the formulation. The pharmaceutical formulation comprises a compound of the invention in combination with one or more pharma- ceutically acceptable ingredients. The excipient may be in the form of a solid, semi-solid or liquid diluent, cream or capsule. These pharmaceutical formulations are further objects of the present invention. Usually, the amount of active compound is 0.1-95% by weight of the formulation, preferably 0.2-20% by weight for parenteral preparations and 1-50% by weight for oral preparations. For clinical use of the method of the present invention, the pharmaceutical composition of the present invention is formulated into a formulation suitable for the intended route of administration.

[0025] The present disclosure is based, at least in part, on the discovery that the combination of hyaluronan-nimesulide conjugates and pyrimidine-based nucleotide analogs, such as fluoropyrimidines, unexpectedly achieves a synergistic effect in suppressing the growth of certain tumors in some mouse models and cell lines, compared to hyaluronan-nimesulide conjugates alone or fluoropyrimidines alone.Therefore, the present disclosure proposes a method for treating locally advanced or metastatic tumors.Some embodiments of the present disclosure relate to a method for treating locally advanced or metastatic tumors.Also provided herein is the use of the hyaluronan-nimesulide conjugates in treating locally advanced or metastatic tumors, and its use in the manufacture of medicaments for the treatment purpose.The medicaments (i.e., pharmaceutical compositions) are of course subject matter within the scope of the present application.

[0026] In one embodiment, the disclosure relates to a method for treating a locally advanced or metastatic tumor in a patient in need of treatment.

[0027] According to some embodiments of the present disclosure, the method includes administering to a patient an effective amount of a hyaluronan-nimesulide conjugate in combination with an effective amount of a pyrimidine-based nucleotide analog, such as a fluoropyrimidine.

[0028] For example, the hyaluronan-nimesulide conjugate can be formulated with a pharmaceutically acceptable excipient into a pharmaceutical composition suitable for desired administration mode.A pharmaceutical composition prepared according to the presently disclosed and claimed inventive concept is a single unit dosage form suitable for parenteral (e.g., subcutaneous, intravenous, bolus injection, intramuscular, or intraarterial) administration to a patient.As can be understood, these pharmaceutical compositions are also within the scope of the present disclosure.

[0029] According to some embodiments of the present disclosure, the locally advanced or metastatic tumor may be a tumor including a locally advanced or metastatic sarcoma, pancreatic tumor, colorectal tumor, liver tumor, melanoma, lung tumor, breast tumor, ovarian tumor, head or neck tumor, gastric tumor, prostate tumor, esophageal tumor, cervical tumor or vaginal tumor, brain tumor (e.g., glioblastoma, astrocytoma, meningioma, or medulloblastoma, etc.). Alternatively, the locally advanced or metastatic tumor may be a liquid tumor including a locally advanced or metastatic multiple myeloma, leukemia, or lymphoma. Additional locally advanced or metastatic solid tumors or liquid tumors are also amenable to treatment using the compositions or methods disclosed herein. As used herein, the term "locally advanced tumor" refers to a cancer that has spread from where it originated in the body to nearby tissues or lymph nodes, but has not spread to other parts of the body. As used herein, the term "metastatic tumor" refers to a cancer that has spread from where it originated in the body to other parts of the body.

[0030] In certain embodiments, the hyaluronan-nimesulide conjugate is administered intravenously to a patient at a Nim equivalent of 0.1-3.2 mg / kg per dose to achieve the desired therapeutic effect. In any embodiment, the hyaluronan-nimesulide conjugate is administered intravenously to a patient at a Nim equivalent of 0.15-2.5 mg / kg per dose, preferably at a Nim equivalent of 0.2-2 mg / kg per dose, more preferably at a Nim equivalent of 0.3-1.5 mg / kg per dose. Specifically, the hyaluronan-nimesulide conjugate is administered intravenously to a patient on designated days of a treatment period (or treatment cycle) at a Nim equivalent of 0.1, 0.15, 0.18, 0.2, 0.25, 0.3, 0.36, 0.4, 0.45, 0.5, 0.54, 0.55, 0.6, 0.65, 0.7, 0.72, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.08, 1.26, 1.44, 1.5, 1.62, 1.8, 2, 2.5, 3, or 3.2 mg / kg. In a particular embodiment, the hyaluronan-nimesulide conjugate is administered to mice at a dose of 2.54-10.16 mg / kg / dose Nim equivalent, and therefore for a human patient, the equivalent dose is 0.205-0.82 mg / kg / dose Nim equivalent.

[0031] In certain embodiments, the hyaluronan-nimesulide conjugate is administered intravenously to a patient at 5-150 mg / kg of hyaluronan-nimesulide conjugate per dose to achieve the desired therapeutic effect. In any embodiment, the hyaluronan-nimesulide conjugate is administered intravenously to a patient at 10-100 mg / kg of hyaluronan-nimesulide conjugate per dose, more preferably 15-50 mg / kg of hyaluronan-nimesulide conjugate per dose. Specifically, the hyaluronan-nimesulide conjugate is administered intravenously to a patient on a designated day of a treatment period (or treatment cycle) at a dose of 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140 or 150 mg / kg of hyaluronan-nimesulide conjugate. In a particular embodiment, the hyaluronan-nimesulide conjugate is administered to a mouse at a dose of 100-400 mg / kg of hyaluronan-nimesulide conjugate, so that for a human patient, the equivalent dose is 8.1-32.4 mg / kg of hyaluronan-nimesulide conjugate.

[0032] In certain embodiments, the hyaluronan-nimesulide conjugate is administered at a certain interval during the treatment period to obtain the desired therapeutic effect.For example, the hyaluronan-nimesulide conjugate is administered once a month, twice a month, three times a month, once every other week, once a week (QW), twice a week (BIW), three times a week (TIW), four times a week, five times a week, six times a week, once every two days (QOD), once a day (QD), twice a day (BID), or three times a day (TID), for a period of about 1 day to about 1 week, about 2 weeks to about 4 weeks, about 1 month to about 2 months, about 2 months to about 4 months, or about 4 months to about 6 months, or longer. In certain embodiments, the hyaluronan-nimesulide conjugate is administered twice (on days 1 and 15) in a 28 day cycle at doses of 0.18, 0.36, 0.72, 0.9, 1.08, 1.44, 2.16, 2.88 and 3.6 mg / kg Nim equivalent.

[0033] According to certain embodiments of the present disclosure, the fluoropyrimidine is 5-FU, 5-fluorocytosine, 5-fluorothymidine, capecitabine, and trifluorothymidine, carmofur, doxifluridine, emitefur, tegafur, or floxuridine. In certain embodiments, the 5-FU is administered to the patient at 1-30 mg / kg / dose. In certain embodiments, the 5-FU is administered intravenously to the patient once a week. In certain embodiments, the hyaluronan-nimesulide conjugate is administered to mice at a dose of 50-200 mg / kg / dose of 5-FU, thus, for a human patient, the equivalent dose is 4.05-16.2 mg / kg / dose of 5-FU.

[0034] In another aspect, the disclosure relates to pharmaceutical compositions in combination with pyrimidines, such as fluoropyrimidines, for treating locally advanced or metastatic tumors.

[0035] According to some embodiments, the pharmaceutical composition comprises an effective amount of a hyaluronan-nimesulide conjugate and a pharma- ceutically acceptable excipient, wherein the pharmaceutical composition is used in combination with a fluoropyrimidine.

[0036] Subject matter encompassed within other aspects of the present disclosure includes the use of a hyaluronan-nimesulide conjugate in the manufacture of a medicament for use in the treatment of locally advanced or metastatic tumors, and a hyaluronan-nimesulide conjugate for use in the treatment of locally advanced or metastatic tumors, wherein the medicament or the hyaluronan-nimesulide conjugate is used in combination with a pyrimidine-based nucleotide analogue, such as a fluoropyrimidine.

[0037] Yet another aspect of the present disclosure relates to the use of hyaluronan-nimesulide conjugates in the manufacture of medicaments in combination with pyrimidine-based nucleotide analogs, such as fluoropyrimidines, for use in the treatment of locally advanced or metastatic tumors. Yet another aspect of the present disclosure relates to the use of hyaluronan-nimesulide conjugates in combination with pyrimidine-based nucleotide analogs, such as fluoropyrimidines, for the treatment of locally advanced or metastatic tumors. Similarly, the hyaluronan-nimesulide conjugates, pyrimidine-based nucleotide analogs, such as fluoropyrimidines, and various dosing regimens (including dosages and dosing intervals) for the various tumors described above are also applicable to these aspects.

[0038] The following examples are provided to illustrate certain aspects of the present invention and to aid those skilled in the art in practicing the present invention. These examples are not to be construed as limiting the scope of the present invention in any manner. Without further elaboration, it is believed that those skilled in the art can utilize the present invention to its fullest extent based on the description herein.

[0039] CA102N is a conjugate of modified nimesulide covalently bound to the sodium salt of hyaluronic acid. Specifically, CA102N is a hyaluronan conjugate having at least one disaccharide unit having the following structure:

[0040] [ka] EXAMPLES

[0041] Example 1

[0042] Combined administration of CA102N and 5-FU synergistically suppresses cell proliferation in HT-29 cells

[0043] In this example, HT-29 cells (colorectal adenocarcinoma) were maintained in high glucose Dulbecco's modified Eagle's medium (DMEM) supplemented with 1% penicillin-streptomycin-neomycin (PSN) antibiotic mixture, 1% L-glutamine, 1% sodium pyruvate, and 10% fetal bovine serum (FBS). All culture reagents were purchased from Invitrogen. Culture was performed at 5% CO 2 Cultures were maintained at 37°C in a humidified atmosphere containing 1 × 10 cells per well. 4 HT-29 cells were seeded in 96-well plates at a density of 100 cells / 100 μL. 0.1 mL of various concentrations of CA102N (equivalent to 0–400 μM nimesulide) and / or 5-FU (0–200 μM) was added to each well, and the cells were cultured at 37°C for 48 h.

[0044] Cell viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction by mitochondrial dehydrogenase. Briefly, the supernatant was removed and 100 μL of MTT solution (0.5 mg / mL in PBS) was added. After further incubation at 37° C. for 4 h, the resulting formazan crystals were dissolved in dimethyl sulfoxide (DMSO) (100 μL) and scanned in a multiwell ELISA reader (SpectraMax® M2e Multimode Plate Reader, Molecular Devices, USA) to measure absorbance at 570 nm. All tests were performed in triplicate and data were expressed as mean ± SD. Relative cell viability (%) was expressed as a percentage of untreated control cells.

[0045] The data summarized in Figure 1A and Figure 1B show that administration of 100, 200, or 400 μM CA102N (equivalent to Nimesulide) plus 50, 150, or 250 μM 5-FU reduces cell viability of HT-29 cells compared to 50, 150, or 250 μM 5-FU alone (Figure 1A) or 100, 200, or 400 μM CA102N alone (Figure 1B). The effect of combined administration of CA102N and 5-FU is dose-dependent. All combined treatment groups showed statistical differences (p<0.05) compared to the individual control groups.

[0046] Example 2

[0047] Combined administration of CA102N and 5-FU synergistically reduces tumor volume in CT26 mice

[0048] Immunocompetent female Balb / C nude mice (6-8 weeks old) were ordered and received from the National Laboratory Animal Center. Animals were housed in microisolator cages on irradiated corncob bedding under 12-h fluorescent lighting cycles under controlled temperature (20-23°C) and humidity (30-70%). Rodent chow and autoclaved acidified water (pH 2.5-3) were provided ad libitum.

[0049] CT26 is an N-nitroso-N-methylurethane (NNMU)-inducible undifferentiated mouse colon carcinoma cell line. CT26 cancer cells were grown in McCoy's 5a culture medium supplemented with 10% (v / v) FBS and 1% PSN antibiotic mixture. The cells were cultured at 175 cm 2 flasks at 37°C and 5% CO 2 The cells were maintained in a humidified incubator at 4 °C and subcultured 2–3 times weekly. On the day of the study, CT26 cells were thawed and prepared for inoculation into mice. The cells were washed with PBS, counted, and cultured at 2 × 10 6Cells were resuspended in cold PBS at a concentration of 10000 viable cells / 0.1 ml. The cell suspension was mixed with an equal volume of Cultrex ECM and kept on ice during transport to the vivarium. Cells for injection were prepared by placing the ECM-cell mixture into a 1 ml chilled Luer-lok syringe fitted with a 26 7 / 8G (0.5 mm x 22 mm) needle. 200 μl of the cell suspension in ECM was injected subcutaneously in the rear flank. The animals were left undisturbed for 7 days before being observed for tumor growth.

[0050] Animals were randomized into treatment groups and then treated with A) vehicle control (PBS) intravenously (IV) twice weekly (BIW), B) 50 mg / kg 5-FU intravenously once weekly (QW), C) 200 mg / kg CA102N intravenously twice weekly (BIW), or D) 200 mg / kg CA102N intravenously twice weekly (BIW) + 50 mg / kg 5-FU intravenously once weekly (QW) (n=6 per group).

[0051] Animals were monitored weekly for palpable tumors or changes in appearance or behavior. Once tumors were palpable, they were measured using calipers. Tumor volume was calculated using the formula: 1 / 2(4π / 3)(L / 2)(W / 2)H, where L is the length of the tumor, W is the width of the tumor, and H is the height of the tumor. Tumor growth inhibition (TGI) was expressed as Mean%Inhibition and was calculated using the formula: [1-(T-T0) / (C-C0)]. * 100% (where C is the tumor volume in vehicle on a particular dosing day, C0 is the tumor volume in vehicle on the first day of dosing, T is the tumor volume in the treatment group on a particular dosing day, and T0 is the tumor volume in the treatment group on the first day of dosing). Data are expressed as mean ± SEM.

[0052] The results summarized in Figure 2 show that the combined administration of 200 mg / kg CA102N and 50 mg / kg 5-FU synergistically enhanced the therapeutic efficacy compared with the combined administration of CA102N or 5-FU alone (*: p<0.05 vs vehicle control; #: p<0.05 vs 5-FU). Specifically, the combined administration of CA102N and 5-FU achieved a TGI rate of approximately 80%, compared with the TGI rate (approximately 70%) of the combined administration of 5-FU alone, suggesting the synergistic therapeutic effect of the combined administration of CA102N and 5-FU.

[0053] Example 3

[0054] Combined administration of CA102N and 5-FU synergistically reduces tumor volume in CR5038 PDX (patient-derived xenograft) mice

[0055] Thirty-two female NOD / SCID mice (6-8 weeks old) were purchased from Jackson Laboratories. Animals were housed in individual HEPA-ventilated cages (Innocage® IVC, Innovive USA) for stabilization. They were kept under 12-h fluorescent lighting cycles under controlled temperature (20-23°C) and humidity (30-70%). Rodent chow and autoclaved acidified water (pH 2.5-3) were provided ad libitum.

[0056] The CR5038 PDX model is associated with metastatic colon adenocarcinoma. Tumor cryovials (CR5038) were thawed and prepared for inoculation into mice. Cells were washed in PBS, counted, and resuspended in cold PBS at a concentration of 96,000 viable cells / 0.1 ml. The cell suspension was mixed with an equal volume of Cultrex ECM and kept on ice during transport to the vivarium. Cells for injection were prepared by injecting the ECM-cell mixture into a 1 ml chilled Luer-lock syringe fitted with a 26 7 / 8G (0.5 mm x 22 mm) needle. The filled syringe was kept on ice to prevent solidification of the ECM. Animals were prepared for injection under standard approved isoflurane anesthesia and shaved prior to injection. Mice were immobilized individually and the injection site was disinfected with an alcohol swab. 0.2 ml of cell suspension in ECM was injected subcutaneously into the posterior flank. The animals were left undisturbed for 7 days and then observed for tumor growth. Animals were monitored weekly for palpable tumors and changes in appearance or behavior. Once tumors were palpable, they were measured using calipers. Tumor volume was calculated using the formula (longest diameter x shortest diameter) 2 ) / 2.

[0057] Animals were randomized into treatment groups and then treated with A) vehicle control (PBS) intravenously twice weekly (BIW), B) 5-FU at 50 mg / kg intravenously once weekly (QW), C) CA102N at 200 mg / kg intravenously twice weekly (BIW), or D) CA102N at 200 mg / kg intravenously twice weekly (BIW) + 5-FU at 50 mg / kg intravenously once weekly (QW) (n=8 per group).

[0058] The results summarized in FIG. 3 show that administration of CA102N alone did not suppress tumor growth compared to the vehicle control group. Surprisingly, administration of CA102N in combination with 5-FU significantly reduced tumor growth compared to all other groups ( * :p<0.05).

[0059] Example 4

[0060] Combined administration of CA102N and 5-FU synergistically reduces tumor volume in HT-29 tumor-bearing mice

[0061] Immunodeficient, Nu / Nu, female athymic nu / nu (nude) mice (5-6 weeks old, LASCO) were purchased from LASCO. Animals were housed in individual HEPA ventilated cages (Innocage® IVC, Innovive USA) for stabilization. They were exposed to fluorescent lighting on a 12-h cycle under controlled temperature (20-23°C) and humidity (30-70%). Rodent chow and water were provided ad libitum.

[0062] HT-29 human colon adenocarcinoma cells (2 × 10 7 ) was implanted subcutaneously into the right flank of each mouse. Once tumors were detectable, palpable, and measurable, primary tumor volumes were measured and recorded every 3 or 4 days. Tumor volume was calculated as 1 / 2(4π / 3)(L / 2)(W / 2)H, where L is the length of the tumor, W is the width of the tumor, and H is the height of the tumor, and tumors were considered to have an average volume of 80-100 mm. 3 When tumor volumes reached approximately 1500 mm, animals were randomly assigned to treatment and control groups and treated with a single intravenous dose. 3 Subjects were withdrawn from the study when body weight loss reached or exceeded 20% (IACUC protocol restrictions).

[0063] Animals were randomized into treatment groups and then administered A) vehicle control (PBS) intravenously three times per week (TIW); B) 5-FU at 50 mg / kg intravenously once per week (QW); C) CA102N at 200 mg / kg intravenously three times per week (TIW); D) CA102N at 200 mg / kg intravenously once every two weeks (Q2W); E) CA102N at 400 mg / kg intravenously once every two weeks (Q2W); F) CA102N at 400 mg / kg intravenously once per week (QW); G) Animals (n=6 per group) were treated with CA102N at 200 mg / kg intravenously once every 2 weeks (Q2W) plus 5-FU at 50 mg / kg intravenously once a week (QW), H) CA102N at 400 mg / kg intravenously once every 2 weeks (Q2W) plus 5-FU at 50 mg / kg intravenously once a week (QW), or I) CA102N at 400 mg / kg intravenously once a week (QW) plus 5-FU at 50 mg / kg intravenously once a week (QW).

[0064] The results summarized in FIG. 4 show that administration of 5-FU (50 mg / kg, QW) or CA102N (200 mg / kg or 400 mg / kg, Q2W, or 400 mg / kg, QW) alone did not inhibit tumor growth compared to the vehicle control group. Surprisingly, administration of CA102N (200 mg / kg or 400 mg / kg, Q2W, or 400 mg / kg, QW) in combination with 5-FU (50 mg / kg, QW) significantly reduced tumor growth compared to the control or each single agent group ( * :p<0.05).

[0065] Moreover, the TGI (tumor growth inhibition rate) in the 5-FU treatment group decreased continuously from D21 to D28 (from about 60% to about 45%). Similarly, the TGI in the CA102N (200 mg / kg Q2W) treatment group was about 60% at D21 and about 50% at D28, the TGI in the CA102N (400 mg / kg Q2W) treatment group was about 65% at D21 and about 50% at D28, and the TGI in the CA102N (400 mg / kg QW) treatment group was about 65% at D21 and about 50% at D28. In other words, in the single treatment group, the TGI on day 28 was only 50%. In contrast, the TGI on day 28 in the combined administration treatment group is relatively high. Specifically, the TGI in the CA102N (200 mg / kg Q2W) + 5-FU treatment group was approximately 75% on D21 and approximately 65% ​​on D28, whereas the TGI in the CA102N (400 mg / kg Q2W) + 5-FU treatment group was approximately 70% on D21 and approximately 70% on D28.

[0066] Example 5

[0067] Combined administration of CA102N and 5-FU synergistically reduces tumor volume in MDA-MB-231 tumor-bearing mice

[0068] Immunodeficient, Nu / Nu, female athymic nu / nu (nude) mice (5-6 weeks old, LASCO) were purchased from LASCO. Animals were housed in individual HEPA ventilated cages (Innocage® IVC, Innovive USA) for stabilization. They were exposed to fluorescent lighting on a 12-h cycle under controlled temperature (20-23°C) and humidity (30-70%). Rodent chow and water were provided ad libitum.

[0069] MDA-MB-231 human breast cancer cells (5 × 10 6 ) was implanted subcutaneously into the right flank of each mouse. Once tumors were detectable, palpable, and measurable, primary tumor volumes were measured and recorded every 3 or 4 days. Tumor volumes were calculated as 1 / 2×L×W. 2 (In the formula, L is length and W is width.) The tumor was 80-100 mm on average.3 When tumor volumes reached approximately 1500 mm, animals were randomly assigned to treatment and control groups and treated with a single intravenous injection. 3 Subjects were withdrawn from the study when body weight loss reached or exceeded 20% (IACUC protocol restrictions).

[0070] After randomization of animals into treatment groups, animals (n=6 per group) were treated with A) vehicle control (PBS) intravenously 3 times per week (TIW), B) 5-FU at 30 mg / kg intravenously 3 times per week (TIW), C) CA102N at 200 mg / kg intravenously 3 times per week (TIW), D) CA102N at 400 mg / kg intravenously 3 times per week (TIW), E) CA102N + 30 mg / kg 5-FU at 200 mg / kg intravenously 3 times per week (TIW) + 5-FU at 30 mg / kg intravenously 3 times per week (TIW), or F) CA102N at 400 mg / kg intravenously 3 times per week (TIW) + 5-FU at 30 mg / kg intravenously 3 times per week (TIW).

[0071] The results summarized in Figure 5 show that 5-FU (30 mg / kg, TIW) or CA102N (200 mg / kg or 400 mg / kg, TIW) alone inhibited tumor growth compared with the vehicle control group. However, the combined administration of CA102N (400 mg / kg, TIW) + 5-FU (30 mg / kg, TIW) significantly reduced tumor growth compared with the control group or the 5-FU single agent treatment group (*: p<0.05).

[0072] Example 6

[0073] Combined administration of CA102N and 5-FU synergistically suppresses cell proliferation of BxPC-3 cells

[0074] In this example, BxPC-3 cells (human pancreatic cancer cells) were maintained in RPMI1640 medium supplemented with 1% PSN antibiotic mixture, 1% L-glutamine, 1% sodium pyruvate, and 10% FBS. All culture reagents were purchased from Invitrogen. Culture was performed at 5% CO 2Cultures were maintained at 37°C in a humidified atmosphere containing 3 × 10 3 BxPC-3 cells were seeded in 96-well plates at a density of cells / 100 μL / well. 0.1 mL of various concentrations of CA102N (equivalent to 0–600 μM nimesulide) with or without 5-FU (0–20 μM) or 5-FU (5–20 μM) alone was added to each well, and cells were cultured at 37°C for 48 h.

[0075] Cell viability was measured using MTT reduction by mitochondrial dehydrogenase. Briefly, the supernatant was removed and 100 μL of MTT solution (0.5 mg / mL in PBS) was added. After further incubation at 0.37° C. for 4 h, the resulting formazan crystals were dissolved in DMSO (100 μL) and the absorbance was measured by scanning at 570 nm using a multi-well ELISA reader (SpectraMax® M2e Multimode Plate Reader, Molecular Devices, USA). All tests were performed in triplicate and data were expressed as mean ± SD. Relative cell viability (%) was expressed as a percentage of untreated control cells.

[0076] The data summarized in Figure 6 show that administration of 150, 300, or 600 μM CA102N + 5, 10, or 20 μM 5-FU (except 300 μM CA102N + 5 μM 5-FU) resulted in a statistically significant decrease in cell viability of BxPC-3 cells compared to administration of 150, 300, or 600 μM CA102N alone (p<0.05). The effect of combined administration of CA102N and 5-FU was dose-dependent.

[0077] Example 7

[0078] Combined administration of CA102N and 5-FU synergistically suppresses cell proliferation of PANC-1 cells

[0079] In this example, PANC-1 cells (pancreatic cancer) were maintained in DMEM medium supplemented with 1% PSN antibiotic mixture, 1% L-glutamine, 1% sodium pyruvate, 0.8% DMSO, and 10% FBS. All culture reagents were purchased from Invitrogen. Culture was performed at 5% CO 2 Cultures were maintained at 37°C in a humidified atmosphere containing 5 × 10 cells / well. 3 PANC-1 cells were seeded in 96-well plates at a density of cells / 100 μL. 0.1 mL of various concentrations of CA102N (equivalent to 0–600 μM nimesulide) with or without 5-FU (0–20 μM) or 5-FU (5–20 μM) alone was added to each well, and cells were cultured at 37°C for 72 h.

[0080] Cell viability was measured using MTT reduction by mitochondrial dehydrogenase. Briefly, the supernatant was removed and 100 μL of MTT solution (0.5 mg / mL in PBS) was added. After further incubation at 37 °C for 4 h, the resulting formazan crystals were dissolved in DMSO (100 μL) and the absorbance was measured by scanning at 570 nm using a multi-well ELISA reader (Varioskan lux thermo Plate Reader, USA). All tests were performed in triplicate and data were expressed as mean ± SD. Relative cell viability (%) was expressed as a percentage of untreated control cells.

[0081] The data summarized in Figure 7 show that administration of 300 or 600 μM CA102N + 5, 10, or 20 μM 5-FU statistically significantly reduced cell viability of PANC-1 cells compared to administration of 300 or 600 μM CA102N alone (p<0.05). Furthermore, administration of low dose (150 μM) CA102N + high dose (20 μM) 5-FU also statistically significantly reduced cell viability of PANC-1 cells compared to administration of low dose CA102N alone (p<0.05). Meanwhile, co-administration of low dose CA102N with low or medium dose (5 or 10 μM) 5-FU also reduced cell viability of PANC-1 cells to some extent compared to administration of low dose CA102N alone. The effect of co-administration of CA102N and 5-FU is dose-dependent.

[0082] Example 8

[0083] Combined administration of CA102N and 5-FU synergistically suppresses cell proliferation of A549 cells

[0084] In this example, A549 cells (lung cancer) were maintained in F12K medium supplemented with 1% PSN antibiotic mixture, 1% L-glutamine, 1% sodium pyruvate, 0.8% DMSO, and 10% FBS. All culture reagents were purchased from Invitrogen. Culture was performed at 5% CO 2 Cultures were maintained at 37°C in a humidified atmosphere containing 5 × 10 cells / well. 3 A549 cells were seeded in 96-well plates at a density of cells / 100 μL. 0.1 mL of various concentrations of CA102N (equivalent to 0–600 μM nimesulide) with or without 5-FU (0–20 μM) or 5-FU (5–20 μM) alone was added to each well, and the cells were cultured at 37°C for 72 h.

[0085] Cell viability was measured using MTT reduction by mitochondrial dehydrogenase. Briefly, the supernatant was removed and 100 μL of MTT solution (0.5 mg / mL in PBS) was added. After further incubation at 37 °C for 4 h, the resulting formazan crystals were dissolved in DMSO (100 μL) and the absorbance was measured by scanning at 570 nm using a multi-well ELISA reader (Varioskan lux thermo Plate Reader, USA). All tests were performed in triplicate and data were expressed as mean ± SD. Relative cell viability (%) was expressed as a percentage of untreated control cells.

[0086] The data summarized in Figure 8 show that administration of low dose (150 μM) CA102N + 5, 10, or 20 μM 5-FU reduced cell viability of A549 cells compared to administration of low dose CA102N alone (p<0.05). Furthermore, administration of medium or high dose (300 or 600 μM) CA102N + high dose (20 μM) 5-FU also statistically significantly reduced cell viability of A549 cells compared to administration of low dose CA102N alone (p<0.05). On the other hand, co-administration of medium or high dose CA102N with low or medium dose (5 or 10 μM) 5-FU only moderately reduced cell viability of A549 cells compared to administration of medium or high dose CA102N alone. The effect of co-administration of CA102N and 5-FU is dose-dependent.

[0087] Example 9

[0088] Combined administration of CA102N and 5-FU synergistically suppresses cell proliferation of MM.1S cells

[0089] In this example, MM.1S cells (multiple myeloma) were maintained in RPMI medium supplemented with 1% PSN antibiotic mixture, 1% L-glutamine, 1% sodium pyruvate, 0.8% DMSO, and 10% FBS. All culture reagents were purchased from Invitrogen. Culture was performed at 5% CO 2Cultures were maintained at 37°C in a humidified atmosphere containing 5 × 10 cells / well. 3 MM.1S cells were seeded in 96-well plates at a density of 10 cells / 90 μL. 90 μL of various concentrations of CA102N (equivalent to 0–600 μM nimesulide) with or without 5-FU (0–20 μM) or 5-FU (5–20 μM) alone was added to each well, and the cells were cultured at 37°C for 72 h.

[0090] Cell viability was then measured using the alamar Blue assay. Briefly, after 72 h of incubation, 20 μL of alamar Blue was added to each well and incubated at 37 °C in the dark for 4 h. The plates were then shaken for 1 min, fluorescence was detected at a fluorescence excitation wavelength of 560 nm, and fluorescence emission was read at 590 nm by scanning with a multi-well ELISA reader. All tests were performed in triplicate, and data were expressed as mean ± SD. Relative cell viability (%) was expressed as a percentage of untreated control cells.

[0091] The data summarized in Figure 9 show that administration of 150, 300, or 600 μM CA102N + 5, 10, or 20 μM 5-FU (except 600 μM CA102N + 5 μM 5-FU) resulted in a statistically significant reduction in cell viability of MM.S1 cells compared to administration of 150, 300, or 600 μM CA102N alone (p<0.05). The effect of combined administration of CA102N and 5-FU is dose-dependent.

[0092] It will be understood that the above description of the embodiments is given by way of example only, and that various modifications may be made by those skilled in the art. The above specification, examples and data provide a complete description of the structure and use of the exemplary embodiments of the invention. Although various embodiments of the invention have been described with a degree of particularity or with reference to one or more individual embodiments, those skilled in the art may make numerous modifications to the disclosed embodiments without departing from the spirit or scope of the invention.

Claims

1. 1. A method for treating a locally advanced or metastatic solid or liquid tumor, comprising: administering to a patient in need of treatment an effective amount of a hyaluronan-nimesulide conjugate and an effective amount of a fluoropyrimidine; A method comprising:

2. 2. The method of claim 1, wherein the solid tumor is a pancreatic tumor, a colorectal tumor, a liver tumor, a melanoma, a lung tumor, a breast tumor, an ovarian tumor, a head or neck tumor, a gastric tumor, a prostate tumor, an esophageal tumor, a cervical or vaginal tumor, or a brain tumor.

3. The method of claim 2, wherein the liquid tumor is multiple myeloma, leukemia, or lymphoma.

4. 2. The method of claim 1, wherein the hyaluronan-nimesulide conjugate is administered intravenously to the patient.

5. 5. The method of claim 4, wherein the hyaluronan-nimesulide conjugate is administered at an equivalent amount of 0.1 to 3.2 mg / kg / dose of nimesulide.

6. 5. The method of claim 4, wherein the hyaluronan-nimesulide conjugate is administered at 5-150 mg / kg / dose of hyaluronan-nimesulide conjugate.

7. 5. The method of claim 4, wherein the hyaluronan-nimesulide conjugate is administered monthly, twice monthly, three times monthly, once every other week, once weekly (QW), twice weekly (BIW), three times weekly (TIW), four times weekly, five times weekly, six times weekly, once every two days (QOD), once daily (QD), twice daily (BID), or three times daily (TID) over the treatment period.

8. 8. The method of claim 7, wherein the treatment period ranges from about 1 day to about 1 week, from about 2 weeks to about 4 weeks, from about 1 month to about 2 months, from about 2 months to about 4 months, or from about 4 months to about 6 months.

9. 5. The method of claim 4, wherein the hyaluronan-nimesulide conjugate is administered twice during a 28 day treatment period.

10. 2. The method of claim 1, wherein the fluoropyrimidine is 5-fluorouracil, 5-fluorocytosine, 5-fluorothymidine, capecitabine, and trifluorothymidine, carmofur, doxifluridine, emitefur, tegafur, or floxuridine.

11. The method of claim 10, wherein the fluoropyrimidine is 5-fluorouracil.

12. 12. The method of claim 11, wherein the 5-fluorouracil is administered at 1 to 30 mg / kg / dose.

13. 12. The method of claim 11, wherein the 5-fluorouracil is administered intravenously to the patient once a week.

14. Use of a hyaluronan-nimesulide conjugate in the manufacture of a medicament in combination with a fluoropyrimidine for the treatment of locally advanced or metastatic solid or liquid tumors.

15. 15. The use according to claim 14, wherein the solid tumor is a pancreatic tumor, a colorectal tumor, a liver tumor, a melanoma, a lung tumor, a breast tumor, an ovarian tumor, a head or neck tumor, a gastric tumor, a prostate tumor, an esophageal tumor, a cervical or vaginal tumor, or a brain tumor.

16. The use according to claim 15, wherein the liquid tumor is multiple myeloma, leukemia, or lymphoma.

17. The use of claim 14, wherein the hyaluronan-nimesulide conjugate is administered intravenously to a patient.

18. The use of claim 17, wherein the hyaluronan-nimesulide conjugate is administered in an amount equivalent to 0.1-3.2 mg / kg / dose of nimesulide.

19. The use of claim 17, wherein the hyaluronan-nimesulide conjugate is administered at 5-150 mg / kg / dose of hyaluronan-nimesulide conjugate.

20. 18. The use of claim 17, wherein the hyaluronan-nimesulide conjugate is administered monthly, twice monthly, thrice monthly, once every other week, once weekly (QW), twice weekly (BIW), three times weekly (TIW), four times weekly, five times weekly, six times weekly, once every two days (QOD), once daily (QD), twice daily (BID), or three times daily (TID) over the treatment period.

21. 21. The use of claim 20, wherein the treatment period ranges from about 1 day to about 1 week, from about 2 weeks to about 4 weeks, from about 1 month to about 2 months, from about 2 months to about 4 months, or from about 4 months to about 6 months.

22. 18. The use of claim 17, wherein the hyaluronan-nimesulide conjugate is administered twice during a treatment period of 28 days.

23. The use according to claim 14, wherein the fluoropyrimidine is 5-fluorouracil, 5-fluorocytosine, 5-fluorothymidine, capecitabine, and trifluorothymidine, carmofur, doxifluridine, emitefur, tegafur, or floxuridine.

24. 24. The use according to claim 23, wherein the fluoropyrimidine is 5-fluorouracil.

25. 25. The use of claim 24, wherein the 5-fluorouracil is administered at 1 to 30 mg / kg / dose.

26. 25. The use of claim 24, wherein the 5-fluorouracil is administered intravenously to the patient once a week.

27. A hyaluronan-nimesulide conjugate for use in combination with a fluoropyrimidine in the treatment of locally advanced or metastatic solid or liquid tumors.

28. The hyaluronan-nimesulide conjugate for use according to claim 27, wherein the solid tumor is a pancreatic tumor, a colorectal tumor, a liver tumor, a melanoma, a lung tumor, a breast tumor, an ovarian tumor, a head or neck tumor, a gastric tumor, a prostate tumor, an esophageal tumor, a cervical or vaginal tumor, or a brain tumor.

29. The hyaluronan-nimesulide conjugate for use according to claim 28, wherein the liquid tumor is multiple myeloma, leukemia, or lymphoma.

30. The hyaluronan-nimesulide conjugate for use according to claim 27, wherein the hyaluronan-nimesulide conjugate is administered intravenously to a patient.

31. The hyaluronan-nimesulide conjugate for use according to claim 30, wherein the hyaluronan-nimesulide conjugate is administered in an amount equivalent to 0.1 to 3.2 mg / kg / dose of nimesulide.

32. The hyaluronan-nimesulide conjugate for use according to claim 30, wherein the hyaluronan-nimesulide conjugate is administered at 5 to 150 mg / kg / dose of the hyaluronan-nimesulide conjugate.

33. 31. The hyaluronan-nimesulide conjugate for use according to claim 30, wherein the hyaluronan-nimesulide conjugate is administered monthly, twice monthly, three times monthly, once every other week, once weekly (QW), twice weekly (BIW), three times weekly (TIW), four times weekly, five times weekly, six times weekly, once every two days (QOD), once daily (QD), twice daily (BID), or three times daily (TID) over the course of a treatment period.

34. The hyaluronan-nimesulide conjugate for use according to claim 33, wherein the treatment period ranges from about 1 day to about 1 week, from about 2 weeks to about 4 weeks, from about 1 month to about 2 months, from about 2 months to about 4 months, or from about 4 months to about 6 months.

35. 31. The hyaluronan-nimesulide conjugate for use according to claim 30, wherein the hyaluronan-nimesulide conjugate is administered twice within a 28 day treatment period.

36. The hyaluronan-nimesulide conjugate for use according to claim 27, wherein the fluoropyrimidine is 5-fluorouracil, 5-fluorocytosine, 5-fluorothymidine, capecitabine, and trifluorothymidine, carmofur, doxifluridine, emitefur, tegafur, or floxuridine.

37. The hyaluronan-nimesulide conjugate for use according to claim 36, wherein said fluoropyrimidine is 5-fluorouracil.

38. The hyaluronan-nimesulide conjugate for use according to claim 37, wherein said 5-fluorouracil is administered at 1 to 30 mg / kg / dose.

39. The hyaluronan-nimesulide conjugate for use according to claim 37, wherein said 5-fluorouracil is administered intravenously to a patient once a week.

40. A pharmaceutical composition for use in combination with a fluoropyrimidine in the treatment of locally advanced or metastatic solid or liquid tumors, comprising an effective amount of a hyaluronan-nimesulide conjugate and a pharma- ceutical acceptable excipient.

41. 41. The pharmaceutical composition for use according to claim 40, wherein the solid tumor is a pancreatic tumor, a colorectal tumor, a liver tumor, a melanoma, a lung tumor, a breast tumor, an ovarian tumor, a head or neck tumor, a gastric tumor, a prostate tumor, an esophageal tumor, a cervical or vaginal tumor, or a brain tumor.

42. 42. The pharmaceutical composition for use according to claim 41, wherein the liquid tumor is multiple myeloma, leukemia, or lymphoma.

43. The pharmaceutical composition for use according to claim 40, wherein the hyaluronan-nimesulide conjugate is administered intravenously to the patient.

44. 44. The pharmaceutical composition for use according to claim 43, wherein said hyaluronan-nimesulide conjugate is administered in an amount equivalent to 0.1-3.2 mg / kg / dose of nimesulide.

45. 44. The pharmaceutical composition for use according to claim 43, wherein the hyaluronan-nimesulide conjugate is administered at 5-150 mg / kg / dose of hyaluronan-nimesulide conjugate.

46. 44. The pharmaceutical composition for use according to claim 43, wherein the hyaluronan-nimesulide conjugate is administered monthly, twice monthly, thrice monthly, once every other week, once weekly (QW), twice weekly (BIW), three times weekly (TIW), four times weekly, five times weekly, six times weekly, once every two days (QOD), once daily (QD), twice daily (BID), or three times daily (TID) over the course of the treatment.

47. 47. The pharmaceutical composition for use according to claim 46, wherein the treatment period ranges from about 1 day to about 1 week, from about 2 weeks to about 4 weeks, from about 1 month to about 2 months, from about 2 months to about 4 months, or from about 4 months to about 6 months.

48. 44. The pharmaceutical composition for use according to claim 43, wherein the hyaluronan-nimesulide conjugate is administered twice within a treatment period of 28 days.

49. 41. The pharmaceutical composition for use according to claim 40, wherein the fluoropyrimidine is 5-fluorouracil, 5-fluorocytosine, 5-fluorothymidine, capecitabine, and trifluorothymidine, carmofur, doxifluridine, emitefur, tegafur, or floxuridine.

50. 50. The pharmaceutical composition for use according to claim 49, wherein said fluoropyrimidine is 5-fluorouracil.

51. 51. The pharmaceutical composition for use according to claim 50, wherein said 5-fluorouracil is administered at 1 to 30 mg / kg / dose.

52. 51. The pharmaceutical composition for use according to claim 50, wherein the 5-fluorouracil is administered intravenously to the patient once a week.

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