Pharmaceutical compositions for the treatment of cancer comprising nanoparticles containing a poorly soluble camptothecin compound and combination therapies thereof

Abstract

The present invention relates to the use of nanoparticles containing a poorly soluble camptothecin compound for the treatment of cancer, and a combination therapy of administering the same in combination with an antitumor agent. The pharmaceutical composition comprising the nanoparticles of the present invention exhibits a synergistic cancer treatment effect when used in combination with an antitumor agent, and thus can be easily used as a cancer treatment drug and a combination therapy formulation.

Description

[Technical Field]

[0001] This patent application claims priority to Korean Patent Application No. 10-2020-0163156, filed with the Korean Intellectual Property Office on November 27, 2020, the entire contents of which are incorporated herein by reference.

[0002] This invention relates to a pharmaceutical composition for treating cancer comprising nanoparticles containing a poorly soluble camptothecin compound, and to combination therapy thereof. [Background Technology]

[0003] Cancer treatments include surgery, radiation therapy, and drug therapy (chemotherapy or anticancer agents). However, malignant tumors cannot be treated with surgery as the primary treatment method, and chemotherapy (alone or in combination) is the only option. Globally used cytotoxic anticancer agents for chemotherapy of malignant tumors include: camptothecin-type anticancer agents, including the DNA topoisomerase inhibitor irinotecan; antimetabolites such as gemcitabine; and microtubule inhibitors such as paclitaxel.

[0004] Irinotecan is known to be a semi-synthetic and water-soluble analogue of the natural alkaloid camptothecin, CPT-11. It inhibits DNA replication by suppressing topoisomerase unwinding of DNA, thereby inhibiting cell proliferation. Irinotecan is currently formulated as Camptosar. TM Irinotecan hydrochloride injection and other aqueous solutions are commercially available. Irinotecan is a prodrug-type anticancer agent that is converted into its active metabolite SN-38 in vivo by carboxyesterase 2 (CES2) to exert its effect. The active SN-38 has approximately 100 to 1000 times higher anticancer activity than irinotecan, but it is difficult to develop into a formulation due to its instability in vivo at pH and its extremely poor solubility. Furthermore, a drawback of irinotecan is its very low conversion rate to SN-38 in vivo (2-8%), resulting in significant patient-to-patient variability (more than 4 times), making it unpredictable and requiring precise dosage calculations (mg / m²) for each patient. 2The exact efficacy or side effects of irinotecan, a cytotoxic anticancer agent, are being investigated. The aforementioned active form SN-38 is a very poorly soluble substance and cannot be dissolved using common solubilization methods; therefore, various solubilization studies and developments are currently underway. Carboxylesterase 2 (CES2), converted from irinotecan to the active form SN-38, exhibits significant expression rate deviations in various tumor tissues. Carboxylesterase 2 is most highly expressed in the small and large intestines, the primary indication for irinotecan, but is almost entirely absent in pancreatic cancer, gallbladder cancer, breast cancer, lung cancer, kidney cancer, and prostate cancer. Therefore, if the active form SN-38 could be directly administered, it is hoped that it could exert anticancer effects in the aforementioned tumor tissues where carboxylesterase 2 is almost absent.

[0005] On the other hand, gemcitabine, as a useful anticancer agent, especially for pancreatic cancer, is currently used as a primary standard treatment for monotherapy. However, its tumor-suppressive efficacy is low, and its effect on prolonging the survival of cancer patients is also limited. Furthermore, the limitation of taxane anticancer agents such as paclitaxel or nab-paclitaxel (albumin-based nanoparticle paclitaxel) lies in the fact that, when used as monotherapy, different tumors exhibit varying sensitivities, thus the anticancer effect is not consistent across different patients. Therefore, to overcome the limitations of monotherapy with various cytotoxic anticancer agents in clinical practice, active research is underway to enhance efficacy and improve safety by improving drug delivery technologies or by combining them with anticancer agents with different mechanisms of action.

[0006] This specification cites numerous academic papers and patent documents throughout, and all citations are clearly indicated. The disclosures in the cited papers and patent documents are incorporated herein by reference in their entirety to more clearly illustrate the level of the technical field to which this invention pertains and the content of this invention. [Summary of the Invention]

[0007] [Technical Issues]

[0008] It is generally believed that chemopreventricular anticancer agents have low antitumor efficacy, not only because of their short residence time in the bloodstream, but also because of their low selective drug delivery efficiency to tumor tissues. Furthermore, due to the high mutation rate of tumors, the sensitivity of each tumor to drugs varies greatly. Therefore, the inventors conducted in-depth research to develop a combination therapy that not only effectively delivers highly active anticancer agents to tumor tissues, but also increases tumor sensitivity by combining antitumor agents with different mechanisms of action, thereby maximizing antitumor efficacy and improving safety. Ultimately, it was confirmed that the combination of antitumor agents with nanoparticle formulations containing both hydrophobic and hydrophilic camptothecin showed excellent antitumor effects, thus completing this invention.

[0009] Therefore, the object of the present invention is to provide a pharmaceutical composition for treating cancer, which is administered in combination with an antitumor agent.

[0010] Another object of the present invention is to provide a pharmaceutical combination formulation comprising, as an active ingredient: particles containing a hydrophobic camptothecin compound, a hydrophilic camptothecin compound, and an amphiphilic block copolymer composed of hydrophobic and hydrophilic blocks; and an antitumor agent, administered simultaneously, alone, or sequentially to treat cancer.

[0011] Solution to the problem

[0012] According to one aspect of the present invention, a pharmaceutical composition for treating cancer is provided, which is administered in combination with an antitumor agent.

[0013] The pharmaceutical composition of the present invention comprises, as an active ingredient, particles containing hydrophobic camptothecin compounds, hydrophilic camptothecin compounds, and amphiphilic block copolymers composed of hydrophobic and hydrophilic blocks.

[0014] In this specification, camptothecin is described as a topoisomerase inhibitor found in the bark and stems of the Camptotheca (Happy tree). It has shown excellent anticancer effects in preclinical stages, but its low solubility has prevented its use. Therefore, many researchers have developed camptothecin analogs to improve its solubility. Currently, three camptothecin derivatives—irinotecan, topotecan, and belotecan—are approved for use in cancer chemotherapy.

[0015] In this specification, the term "hydrophobicity" refers to a tendency in nonpolar substances to aggregate and be repelled by water molecules. When a hydrophobic substance is in a hydrophilic liquid, it will increase hydrophobic bonds as if it were afraid of water, thus causing the hydrophobic substance to aggregate together.

[0016] In this specification, the term "hydrophilicity" primarily refers to a tendency found in polar substances, indicating a strong affinity for and easy dissolution of polar solvents such as water. For example, the micelle surface of hydrophilic polymers or surfactants exhibits strong hydrophilicity.

[0017] In one embodiment of the present invention, the aforementioned hydrophobic camptothecin compound is selected from at least one of the group consisting of SN-38 (7-ethyl-10-hydroxycamptothecin), camptothecin, 10-hydroxycamptothecin, and pharmaceutically acceptable salts thereof, but is not limited thereto.

[0018] In another embodiment of the present invention, the aforementioned hydrophilic camptothecin compound is selected from at least one of irinotecan, topotecan, belootecan, exatecan, lurtotecan, sinotecan, rubitecan, 9-nitrocamptothecin, 9-aminocamptothecin, gimatecan, carrenitecin, silatecan, diflomotecan, elomotecan, their pharmaceutically acceptable salts or their glucuronide metabolites, and the glucuronide metabolites of the aforementioned hydrophobic camptothecin compounds, but is not limited thereto. The aforementioned hydrophilic camptothecin compounds generally include compounds classified as hydrophobic drugs, but the term "hydrophilic" should be understood as referring to the relative hydrophilicity of the components constituting the particles of the present invention compared to the aforementioned hydrophobic camptothecin compounds. That is, the aforementioned hydrophilic camptothecin compounds refer to camptothecin compounds that have relative hydrophilicity compared to the hydrophobic camptothecin compounds contained in the particles of the present invention.

[0019] According to specific examples of the present invention, the hydrophobic camptothecin compounds constituting the particles of the present invention may be camptothecin, SN-38, or mixtures thereof, and the hydrophilic camptothecin compounds may be irinotecan hydrochloride, topotecan hydrochloride, or glucuronide analogs of SN-38.

[0020] In this specification, the term "copolymer" refers to a polymer made from two or more different monomers. For example, when styrene and acrylonitrile react in the same reaction vessel, a copolymer containing both monomers is formed. A "block copolymer" refers to a copolymer with one monomer block linked to another. The case where a block of substance A is followed by a block of substance B is represented as -[-AB-]-. If the chain consists of only one chain of each monomer, it is called AB type; if there is a block of substance B in the middle and blocks of substance A at both ends, it is called ABA type; if there are three different blocks in the main chain, it is called ABC type. Block copolymers are mainly formed through ionic polymerization. Unlike other copolymers, block copolymers possess many of the physical properties of homogeneous polymers made from two monomers.

[0021] In one embodiment of the present invention, the amphiphilic block copolymer constituting the particles of the present invention is composed of AB or ABA blocks. Wherein, A is a hydrophilic polymer selected from, but not limited to, monomethoxy polyethylene glycol, dimethoxy polyethylene glycol, polyethylene glycol, polypropylene glycol, monomethoxy polyethylene glycol, polyethylene oxide, and polyacrylic acid.

[0022] Furthermore, the aforementioned B is a hydrophobic polymer, specifically polylactic acid, poly-L-lactide, poly-D-lactide, poly-D,L-lactide, poly(lactide-co-glycolide), polyglyconic acid, polyglycolide, polylactic acid-gluconic acid copolymer, polymandelic acid, polycaprolactone, polydioxan-2-one, polyglutamic acid, and polyaspartic acid. (acid), polyornithine, polyorthoesters, derivatives thereof, or at least one of these compounds, but not limited thereto, and it will be apparent to those skilled in the art that any compound capable of forming an amphiphilic block copolymer available in the art may be used without limitation.

[0023] In specific embodiments of the present invention, the aforementioned amphiphilic block copolymers are: methoxy polyethylene glycol-polylactic acid (mPEG-PDLLA, mPEG-Poly(D,L)Lactic Acid); polyethylene glycol-polycaprolactone (PEG-PCL, [poly(ethylene glycol)-b-poly(carprolactone)]); polyethylene glycol-polylactic acid (PEG-PLA, [poly(ethylene glycol)-b-poly(lactic acid)]); methoxy polyethylene glycol-polyglycolic acid (mPEG-PGA, [monomethoxy poly(ethylene glycol)-b-poly(glycolic acid)]); methoxy polyethylene glycol-polyglycolic acid-glycolic acid (mPEG-PLGA, [monomethoxy poly(ethylene glycol)-b-poly(lactide-co-glycolide)]); and polyethylene glycol-polyaspartic acid benzyl ester (PEG-PBLA, [poly(ethylene glycol)-b-poly(β-benzyl-L-aspartic acid)]). [poly(ethyleneglycol)-b-poly(glutamic acid)]); [poly(ethyleneglycol)-aspartic acid]; and / or [poly(ethyleneglycol)-b-poly(aspartic acid)]; and / or [poly(ethylene glycol)-PLA-PEG];

[0024] According to the most specific example of the present invention, the characteristics of the particles of the present invention and the preparation method thereof are described in detail in the applicant’s Korean Patent No. 10-2094543, and the matters disclosed in the above patent are incorporated herein by reference.

[0025] The particles containing SN-38 as a hydrophobic camptothecin compound, irinotecan hydrochloride as a hydrophilic camptothecin compound, and mPEG-PDLLA as an amphiphilic block copolymer, prepared according to the above-mentioned authorized patent or the following embodiments, are named SNB-101.

[0026] In another embodiment of the present invention, the weight ratio of the hydrophobic camptothecin compound to the hydrophilic camptothecin compound constituting the particles of the present invention is 1-10:1-10, 1-10:1-5, 1-10:1-3, 1-10:1, 1-5:1-10, 1-3:1-10, 1:1-10, specifically 1-5:1-5, 1-5:1-3, 1-5:1, 1-3:1-5, 1:1-5, more specifically 1-3:1-3, 1-3:1, 1:1-3, but not limited thereto.

[0027] In specific examples of the present invention, the weight ratio of the above-mentioned hydrophobic camptothecin compound to the above-mentioned hydrophilic camptothecin compound can be 1:1-10, 1:1-5, 1:1-3 or 1:1-2, most specifically, it can be 1:1.59, but is not limited thereto.

[0028] Furthermore, in one embodiment of the present invention, the weight ratio of the sum of (a) the hydrophobic camptothecin compound and the hydrophilic camptothecin compound to (b) the amphiphilic block copolymer is 1:0.1-200, 1:0.5-200, 1:1-200, 1:2-200, 1:5-200, 1:10-200, 1:50-200, 1:100-200, 1:150-200, 1:0.1-100, 1:0.5-100, 1:1-100, 1:2-100, 1:5-100, 1:10-100, 1:20-100, 1: 50-100, 1:0.1-50, 1:0.5-50, 1:1-50, 1:5-50, 1:10-50, 1:20-50, 1:0.1-20, 1:0.5-20, 1:1-20, 1:5-20, 1:10-20, 1:0.1-10, 1:0.5-10 or 1:1-10, but not limited to these.

[0029] As used in this article, the terms “to,” “-,” or “~” used to describe the interval between two numerical values ​​refer to the range between the values, including the values ​​recorded before and after them.

[0030] In one embodiment of the present invention, the particles of the present invention are prepared by the following method: First, hydrophobic camptothecin (e.g., SN-38) and hydrophilic camptothecin (e.g., irinotecan hydrochloride) are placed together in an organic solvent and stirred until completely dissolved. An amphiphilic block copolymer (e.g., mPEG-PDLLA) pre-dissolved in the organic solvent is added while stirring. The mixture is dried using a rotary evaporator or a vacuum dryer. An aqueous solvent (e.g., distilled water, phosphate buffered saline (PBS)) is added to the residue, and ultrasonic waves are applied to an ultrasonic cleaner to prepare the particles of the present invention.

[0031] In one embodiment of the present invention, the aforementioned organic solvent is a C1 to C5 alcohol (methanol, ethanol, propanol, butanol, n-butanol, isopropanol, 1-pentanol, 2-butoxyethanol, isobutanol, etc.), alkyl acetate, acetone, acetonitrile, chloroform, benzene, toluene, xylene, acetone, fluorocarbons, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1-pentene, 1-chlorobutane, 1-chloropentane, diisopropyl ether, 2-chloropropane, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, tetrahydrofuran (THF), or a mixture thereof, but is not limited thereto.

[0032] The pharmaceutical compositions of the present invention comprise antitumor agents.

[0033] The aforementioned antitumor agents include at least one selected from the group consisting of taxane anticancer agents, albumin-bound taxane anticancer agents, vascular endothelial growth factor (VEGF) inhibitors, and gemcitabine.

[0034] In one embodiment of the present invention, the aforementioned taxane anticancer agent may be at least one selected from the group consisting of paclitaxel, docetaxel, larotaxel, cabazitaxel, and their pharmaceutically acceptable salts, but is not limited thereto.

[0035] In one embodiment of the present invention, the aforementioned albumin-bound taxane anticancer agent may be at least one selected from the group consisting of albumin-bound paclitaxel (albumin nanoparticle paclitaxel or nab-paclitaxel) and albumin-bound docetaxel (albumin nanoparticle docetaxel or nab-docetaxel), but is not limited thereto.

[0036] In one embodiment of the present invention, the vascular endothelial growth factor (VEGF) inhibitor may be at least one selected from the group consisting of bevacizumab, ranibizumab, and aflibercept, but is not limited thereto.

[0037] In a specific example of the present invention, the hydrophobic camptothecin compound is SN-38, the hydrophilic camptothecin compound is irinotecan hydrochloride, and the amphiphilic block copolymer is methoxy polyethylene glycol-polylactic acid (mPEG-PDLLA, mPEG-Poly(D,L)Lactic Acid).

[0038] The inventors have named the particles prepared according to the above specific examples as SNB-101.

[0039] The stable nanoparticle formulation of SN-38 and irinotecan hydrochloride (SNB-101) is a formulation that solubilizes and formulates the extremely poorly soluble SN-38. It can directly deliver SN-38, which has excellent anticancer effects but cannot be utilized due to low solubility. Therefore, it can show anticancer effects against pancreatic cancer, ovarian cancer, lung cancer, breast cancer and other cancers that lack carboxylesterase. It can also be expected to have effects on new indications.

[0040] Based on SN-38, the particle content of the active ingredient in the pharmaceutical composition of the present invention, as a specific example of the present invention, is preferably 0.1 to 1 mg / ml, more preferably 0.2 to 1.0 mg / ml, relative to the particle composition. Furthermore, the irinotecan hydrochloride content is preferably 0.1 to 2 mg / ml, more preferably 0.2 to 1.6 mg / ml, relative to the particle composition described above.

[0041] In one embodiment of the present invention, the particles have a dual structure. The dual structure includes: an internal structure composed of the aforementioned hydrophilic camptothecin compound and a hydrophobic camptothecin compound; and an external structure composed of the aforementioned amphiphilic block copolymer.

[0042] In the preparation of the particles with the above-mentioned dual structure of the present invention, a hydrophilic camptothecin compound and a hydrophobic camptothecin compound are first mixed and dissolved in an organic solvent. An amphiphilic polymer (amphiphilic block copolymer) dissolved in the organic solvent is then added to the mixture while stirring and drying. The dried material is then emulsified in an aqueous solvent by ultrasonic treatment or other methods to form water-dispersible nanoparticles.

[0043] It is known that amphiphilic block copolymers, in which hydrophilic and hydrophobic blocks are combined in a specific ratio, self-assemble in aqueous solutions to form micelle-like aqueous dispersible particles. Therefore, it is presumed that the particles of the present invention contain the aforementioned hydrophilic and hydrophobic camptothecin compounds within the aqueous dispersible particles, and that the aforementioned amphiphilic block copolymer forms an outer shell. More specifically, in the structure of the aqueous dispersible particles of the present invention, it is presumed that the hydrophobic blocks in the aforementioned amphiphilic block copolymer face the relatively hydrophobic internal structure composed of camptothecin compounds, while the hydrophilic blocks face the external aqueous solvent, thereby forming a shell.

[0044] In one embodiment of the present invention, the above-described dual-structure water-dispersible particles of the present invention are characterized in that they spontaneously form particles when dispersed in an aqueous solution.

[0045] In another embodiment of the present invention, the average diameter of the particles is 2 to 200 nm. When the size of the particles prepared as described in the present invention is less than 200 nm, non-selective removal of the reticuloendothelial system (RES system) in vivo can be avoided. Therefore, it is preferable to prepare particles with a homogeneous particle size of less than 200 nm.

[0046] In one embodiment of the present invention, the above-mentioned water-dispersible particles of the present invention can be mixed with freeze-drying protectants such as trehalose and mannitol and then freeze-dried.

[0047] The average diameter (particle size) of the particles described above in this invention is approximately 200 nm or less. This allows for the utilization of the enhanced permeation and retention (EPR) effect in tumor tissue to maximize the delivery of the aforementioned drugs to tumor tissue. The enhanced permeation and retention effect disrupts the normal integrity of the vascular system (especially capillaries), causing leakage of nanoscale particle delivery carriers (nanoparticles, liposomes, micelles) from the capillaries and allowing the delivery carrier to deposit at the tumor site.

[0048] Therefore, due to this characteristic of the particles (SNB-101) of the present invention, SN-38 and irinotecan contained in SNB-101 can remain in the bloodstream for a prolonged period, thereby promoting drug accumulation in tumor tissue. Therefore, it is expected that by increasing the drug exposure to cancer cells within the tumor and reducing the drug exposure to normal tissues, safety can be improved, and the maximum tolerated dose (MTD) of the drug can be increased.

[0049] In one embodiment of the present invention, a pharmaceutical composition according to one aspect of the present invention is used to treat cancer. The cancers described herein are selected from, but are not limited to, the group consisting of, gastric cancer, ovarian cancer, uterine cancer, cervical cancer, small cell lung cancer, non-small cell lung cancer, pancreatic cancer, breast cancer, esophageal cancer, retinoblastoma, oral cancer, salivary gland cancer, laryngeal cancer, pharyngeal cancer, rectal cancer, colon cancer, colorectal cancer, kidney cancer, prostate cancer, melanoma, liver cancer, gallbladder and other biliary tract cancers, thyroid cancer, bladder cancer, brain cancer and central nervous system cancers, bone tumors, skin cancer, non-Hodgkin's lymphoma, and Hodgkin's lymphoma. The brain cancers described herein may be gliomas, meningiomas, schwannomas, pituitary adenomas, metastatic brain tumors, or skull base tumors. The gliomas described herein include astrocytomas, oligodendrogliomas, ependymomas, or mixtures thereof.

[0050] In a specific example of the present invention, the cancer that is the target disease for treatment of the above-described pharmaceutical composition is pancreatic cancer, gastric cancer, breast cancer, lung cancer, colorectal cancer, or a combination thereof.

[0051] When the particles or compositions comprising the present invention are prepared into pharmaceutical compositions, the pharmaceutical compositions of the present invention may contain pharmaceutically acceptable carriers. The aforementioned pharmaceutically acceptable carriers are those commonly used in formulations, including, but not limited to, lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylparaben, propylparaben, talc, magnesium stearate, and mineral oil. In addition to the above-mentioned components, the pharmaceutical compositions of the present invention may also include lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc. Suitable pharmaceutically acceptable carriers are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0052] In specific examples of the present invention, the pharmaceutical composition further includes sucrose, mannitol, sorbitol, glycerol, trehalose, polyethylene glycol excipients, and cyclodextrin excipients (α, β, γ-cyclodextrin, hydroxycyclodextrin to cyclodextrin derivatives, etc.). These excipients are added to the particles, which are the active ingredient of the pharmaceutical composition, to function as cryoprotectants or osmotic pressure regulators, and are formulated into dosage forms by methods such as freeze-drying and solvent evaporation.

[0053] The pharmaceutical composition of the present invention can be administered orally or via parenteral route. In the case of parenteral administration, it can be administered via intravenous administration, intra-arterial administration, intrarectal administration, subcutaneous administration, intradermal administration, intramuscular administration, intranasal administration, intramucosal administration, intradural administration, intraperitoneal administration, intraocular administration, etc. Specifically, it can be administered intravenously.

[0054] The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration route, patient's age, weight, sex, disease state, food, administration time, route of administration, excretion rate, and response sensitivity. Generally, a skilled physician can easily determine and prescribe the necessary effective dosage for treatment or prevention. According to a specific example of the present invention, the daily dosage of the pharmaceutical composition is 0.001-100 mg / kg.

[0055] In a specific embodiment of the present invention, the dosage and frequency of administration of SN-38 and irinotecan hydrochloride contained in SNB-101 of the present invention can be based on SN-38, with a daily dose of 1-5 mg / m². 2 It can be administered in one to several doses. For example, in the case of an injectable formulation, it can be administered intravenously in one to three doses to a human patient (based on a body weight of 60 kg and a body surface area of ​​1.67 m²). 2 The daily dosage is 1 to 100 mg, specifically 20 mg to 85 mg, and more specifically 30 mg to 85 mg. However, the dosage and frequency of administration are not limited to these.

[0056] The pharmaceutical compositions of the present invention can be formulated using pharmaceutically acceptable carriers and / or excipients through methods readily practiced by those skilled in the art, thereby preparing them in unit dose form or in multi-volume containers. In this case, the dosage form can be a solution, suspension, or emulsion in an oily or aqueous medium, or it can be a concentrate, powder, granules, tablet, or capsule, and may further include dispersants or stabilizers.

[0057] The pharmaceutical compositions of the present invention can be administered simultaneously with known compounds or pharmaceutical compositions that have cancer therapeutic effects.

[0058] In one embodiment of the present invention, the aforementioned known compound or pharmaceutical composition comprises at least one selected from the group consisting of taxane anticancer agents, albumin-bound taxane anticancer agents, vascular endothelial growth factor (VEGF) inhibitors, and gemcitabine.

[0059] According to one aspect of the present invention, the present invention provides a cancer treatment method comprising: administering (a) a first pharmaceutical composition comprising particles containing a hydrophobic camptothecin compound, a hydrophilic camptothecin compound, and an amphiphilic block copolymer composed of hydrophobic and hydrophilic blocks, and (b) a second pharmaceutical composition comprising an antitumor agent as an active ingredient, or in combination with an antitumor therapy, to a subject requiring administration.

[0060] In one embodiment of the present invention, the first pharmaceutical composition and the second pharmaceutical composition are administered simultaneously, individually, or sequentially.

[0061] In one embodiment of the present invention, the first pharmaceutical composition and the second pharmaceutical composition are administered as a combined formulation or a single formulation.

[0062] In one embodiment of the present invention, the above-mentioned antitumor therapy includes surgical therapy, radiotherapy, or a combination thereof.

[0063] In one embodiment of the present invention, the cancers mentioned above are selected from the group consisting of gastric cancer, ovarian cancer, uterine cancer, cervical cancer, small cell lung cancer, non-small cell lung cancer, pancreatic cancer, breast cancer, esophageal cancer, retinoblastoma, oral cancer, salivary gland cancer, laryngeal cancer, pharyngeal cancer, rectal cancer, colon cancer, colorectal cancer, kidney cancer, prostate cancer, melanoma, liver cancer, gallbladder and other biliary tract cancers, thyroid cancer, bladder cancer, brain cancer, central nervous system cancer, bone tumors, skin cancer, non-Hodgkin's lymphoma, and Hodgkin's lymphoma, but are not limited thereto. The aforementioned brain cancers can be gliomas, meningiomas, schwannomas, pituitary adenomas, metastatic brain tumors, or skull base tumors. The aforementioned gliomas include astrocytomas, oligodendrogliomas, ependymomas, or mixed types thereof.

[0064] In a specific embodiment of the present invention, the aforementioned cancer is pancreatic cancer, gastric cancer, breast cancer, lung cancer, colorectal cancer, or a combination thereof.

[0065] According to another aspect of the present invention, the present invention provides a pharmaceutical combination formulation comprising, as an active ingredient: particles containing a hydrophobic camptothecin compound, a hydrophilic camptothecin compound, and an amphiphilic block copolymer composed of hydrophobic and hydrophilic blocks; and an antitumor agent.

[0066] The various active ingredients of the above-mentioned drug combination preparation are administered simultaneously, individually, or sequentially to treat cancer.

[0067] According to another aspect of the present invention, the present invention provides a kit for treating cancer, wherein the first pharmaceutical composition comprises particles containing a hydrophobic camptothecin compound, a hydrophilic camptothecin compound, and an amphiphilic block copolymer composed of hydrophobic and hydrophilic blocks; and the second pharmaceutical composition comprises an antitumor agent.

[0068] The first and second pharmaceutical compositions of the above-mentioned kit for treating cancer are administered simultaneously, alone, or sequentially to treat cancer.

[0069] In one embodiment of the present invention, the first pharmaceutical composition and the second pharmaceutical composition are administered as a combined formulation or a single formulation.

[0070] As used in this specification, the terms "administration" or "giving" mean giving a therapeutically effective amount of the composition of the present invention directly to a subject (individual) suffering from cancer, so that the same amount is formed in the subject's body.

[0071] The "therapeutic effective amount" of the above-described composition refers to the amount of composition sufficient to provide a therapeutic or preventive effect to the subject to whom the composition is to be administered, and includes the meaning of "preventive effective amount". Furthermore, the term "subject" as used in this specification includes, but is not limited to, humans, mice, rats, guinea pigs, dogs, cats, horses, cattle, pigs, monkeys, chimpanzees, baboons, or rhesus monkeys. Specifically, the subject of this invention is a human.

[0072] The cancer treatment methods, pharmaceutical combinations, and kits for treating cancer described above in this invention contain the same active ingredients as the pharmaceutical compositions for treating cancer in one aspect of this invention, and use antitumor agents as combination formulations. Therefore, the content described in one aspect of this invention is equally applicable to repeated content.

[0073] [The effects of the invention]

[0074] This invention relates to the use of nanoparticles containing poorly soluble camptothecin compounds for the treatment of cancer, and to a combination therapy of administering them in combination with taxane antitumor agents. Pharmaceutical compositions containing the nanoparticles of this invention exhibit synergistic cancer therapeutic effects when used in combination with taxane antitumor agents, and therefore can be readily used as cancer treatment agents and combination therapy formulations. [Attached Image Description]

[0075] Figure 1 The changes in body weight of mice in a pancreatic cancer model after administration of AsPC-1 transplanted subcutaneously are shown.

[0076] Figure 2The tumor volume change demonstrated the tumor-suppressive effect of treating subcutaneously transplanted AsPC-1 pancreatic cancer model mice with the experimental substance.

[0077] Figure 3 The tumor-suppressive effect of treating subcutaneously transplanted AsPC-1 pancreatic cancer model mice with changes in tumor weight was demonstrated by the experimental substance.

[0078] Figure 4 Photograph of a tumor removed on day 22 after a mouse model of pancreatic cancer with subcutaneous transplantation of AsPC-1 was treated with the experimental substance.

[0079] Figure 5 The changes in body weight of mice after administration of Hs746T to a gastric cancer model mouse with subcutaneous transplantation are shown.

[0080] Figure 6 The tumor volume change demonstrated the tumor-suppressive effect of treating subcutaneously transplanted Hs746T gastric cancer model mice with the experimental substance.

[0081] Figure 7 The tumor-suppressive effect of treating subcutaneously transplanted Hs746T gastric cancer model mice with changes in tumor weight was demonstrated by the experimental substance.

[0082] Figure 8 Photograph of a tumor removed on day 22 after a mouse model of gastric cancer with subcutaneous transplantation of Hs746T was treated with the experimental substance.

[0083] Figure 9 The tumor volume change demonstrated the tumor-suppressive effect of treating mice with a breast cancer model subcutaneously transplanted with MDA-MB231 using the experimental material.

[0084] Figure 10 The changes in body weight of mice with a breast cancer model after subcutaneous transplantation of MDA-MB231 with the test substance are shown.

[0085] Figure 11 The tumor volume change demonstrates the tumor-suppressive effect of treating subcutaneously transplanted A549 lung cancer model mice with the experimental substance.

[0086] Figure 12 The changes in body weight of mice with a lung cancer model after subcutaneous transplantation of A549 are shown.

[0087] Figure 13 The changes in tumor volume in a colorectal cancer model mouse after subcutaneous transplantation of HT-29 with the test substance are shown.

[0088] Figure 14 The changes in tumor volume in a small cell lung cancer model mouse after subcutaneous transplantation of NCI-H69 with the test substance are shown.

[0089] Figure 15 The image shows a photograph of a small cell lung cancer model mouse after subcutaneous transplantation of NCI-H69 with the experimental substance.

[0090] Figure 16 The changes in tumor volume in a colorectal cancer model mouse after subcutaneous transplantation of HCT116 with the experimental material are shown.

[0091] Figure 17 The image shows a photograph of a colorectal cancer model mouse after treatment with the experimental substance following subcutaneous transplantation of HCT116.

[0092] Figure 18 The changes in tumor volume in a mouse model of non-small cell lung cancer after subcutaneous transplantation of A549 with the test substance are shown.

Detailed Implementation Methods

[0093] The present invention will now be described in more detail through embodiments. These embodiments are merely for the purpose of describing the invention more specifically, and it will be apparent to those skilled in the art that the scope of the invention is not limited to these embodiments in accordance with the spirit of the invention.

[0094]

Example

[0095] Throughout the instruction manual, unless otherwise stated, the percentage used to indicate the concentration of a particular substance is (weight / weight)% for solid / solid, (weight / volume)% for solid / liquid, and (volume / volume)% for liquid / liquid.

[0096] [Example 1: Preparation of the nanoparticles (SNB-101) of the present invention]

[0097] The inventors placed 20 mg of SN-38 (7-ethyl-10-hydroxycamptothecin), the hydrophobic camptothecin, and 30 mg of irinotecan hydrochloride (trihydrate), the hydrophilic camptothecin, into a round-bottom flask. 10 ml of acetonitrile was added, and the mixture was sonicated using a sonicator to completely dissolve both drugs. Then, 100 mg of methoxy polyethylene glycol-poly(D,L,lactic acid) [mPEG-Poly(D,L)Lactic Acid] (mPEG-PDLLA) was weighed into the round-bottom flask, and 10 ml of organic solvent (50:50 v / v mixture of ethanol and acetonitrile) was added. The mixture was stirred for 30 minutes until completely dissolved. The completely dissolved mPEG-PDLLA solution was then slowly added in several portions to the solution containing SN-38 and irinotecan hydrochloride, with stirring for 10–15 seconds after each addition. After mixing the above solution in a round-bottom flask, the mixture is dried in a rotary evaporator under reduced pressure to completely remove the organic solvent, yielding a thin film. 20 ml of sterile distilled water is then added, and ultrasonic waves are applied to the film in an ultrasonic cleaning agent to completely dissolve it.

[0098] The average particle size of the prepared dual-structure nanoparticles was determined by diluting the sample with distilled water to prepare SN-38 at a concentration of 1 mg / ml. The intensity weight-averaged diameter was then measured using a Zetasizer Nano System from Malvern (UK) via dynamic light scattering.

[0099] Then, 200 mg of trehalose and 300 mg of mannitol were added to 20 ml of sterile distilled water as freeze-drying protectants and stirred until completely dissolved.

[0100] Finally, all the freeze-drying protectant solutions were added to the nanoparticle solution of the mixed drug, stirring for 10–15 seconds each time. The final mixture was filtered through a 0.22 μm cellulose acetate membrane filter and filled into packaging vials. The filled vials were freeze-dried to obtain a dry powder or cake-like finished product (named SNB-101).

[0101] [Example 2: The effect of combined drug delivery of the nanoparticles (SNB-101) of the present invention on pancreatic cancer]

[0102] 【2-1. Experimental Materials】

[0103] The substances used in the experiment included: 5% glucose injection and irinotecan hydrochloride (Calmtop Injection). TM CJ Healthcare, South Korea; Nab-paclitaxel; Abraxane TM Abraxis BioScience, LLC, and a nanoparticle composition comprising SN-38 and irinotecan hydrochloride (named SNB-101). The aforementioned irinotecan hydrochloride (CalmtopInjection) TM The dose was 100 mg / 5 mL, purchased from CJ Healthcare. The dose of Abercrombie & Fitch was purchased from Abercrombie & Fitch Biosciences, LLC (USA), and was a lyophilized injectable at a dose of 100 mg / vial.

[0104] The nanoparticle composition (SNB-101) containing SN-38 and irinotecan hydrochloride used in this invention is an injectable preparation. It is a stable nanoparticle dosage form, and its preparation method is described in Korean Patent No. 10-2094543. Each vial of the above-mentioned SNB-101 intravenous injection contains 10 mg of SN-38 and 15.9 mg of irinotecan hydrochloride. It can be prepared by diluting with 9.64 mL of 5% glucose injection (United States Pharmacopeia Standard, USP), and the infusion time can be 90 minutes.

[0105] 【2-2. Experimental Methods】

[0106] (1) Culture AsPC-1 cells

[0107] AsPC-1 was used as the pancreatic cancer cell line for a pancreatic cancer xenograft model. AsPC-1 is known to be an invasive cell line resistant to gemcitabine. [The text abruptly shifts to a seemingly unrelated topic:] ...at 175 cm... 2AsPC-1 cells were cultured in flasks using Roswell Park Memorial Institute medium (RPMI-1640) containing 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin. FBS, RPMI-1640, and penicillin-streptomycin were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and used in the experiments. Cells were cultured at 37°C in 5% CO2 medium. Each cancer cell was refeeded 2-3 times per week, washed with phosphate-buffered saline (PBS; pH 7.4), and then separated from adherent cells using 0.05% trypsin-0.02% EDTA. After centrifuging the isolated cells (3 minutes, 1500 rpm), the culture medium was placed into the accumulated cancer cells and thoroughly mixed with a pipette until evenly dispersed. The cells were then used for subculture and experiments.

[0108] (2) Induction of a mouse model of pancreatic cancer xenograft

[0109] The animals used in this experiment were male BALB / c nu / nu mice (approximately 5 weeks old, with an average weight of 19g ± 20%), widely used in xenograft animal models, which underwent an acclimatization process of about 5-7 days after purchase. On the day of inoculation, the cultured cancer cells were washed with phosphate-buffered saline (PBS, pH 7.4), and then adherent cells were separated using 0.05% trypsin-0.02% EDTA. The separated cells were centrifuged (3 minutes, 1500 rpm), diluted with phosphate-buffered saline, and then injected at 5 × 10⁻⁶ cells per mouse. 6 One cell / 0.2 mL was subcutaneously injected into the right rib of the mouse to confirm that the tumor cell suspension did not flow out from the injection site.

[0110] [Group composition and dosage settings]

[0111] With 200±20mm 3 Mice were grouped based on average pancreatic cancer tumor volume. The composition and dosage of the control and experimental groups are as follows:

[0112] Table 1

[0113]

[0114] * Dosage of SNB-101 labeled with SN-38 / irinotecan hydrochloride

[0115] (3) Administration site and administration method

[0116] The test substance was administered intravenously (iv) to each individual based on their pre-administration body weight. The test substance was administered once a week (on days 0, 7, 14, 17, and 21) for a total of 5 weeks, and dissection was performed on day 22.

[0117] (4) Observation and inspection items

[0118] Clinical symptoms were observed once daily during the experiment, and tumor volume was measured three times weekly while the medication was administered. At the end of the experiment, animals were anesthetized with isoflurane and bled, and tumor tissue was removed from each individual. The removed tumors were weighed and photographed. Tumor volume was measured using vernier calipers (Mitutoyo Corp., Model No.: CD-15CPX, Japan), measuring the major and minor axes of the tumor, and the tumor volume was calculated according to Equation 1 below.

[0119] Formula 1

[0120] Tumor volume = (minor axis) 2 ×major axis × 0.5

[0121] (5) Statistical Analysis

[0122] The experimental results were statistically analyzed using a commercial statistical program (IBM SPSS statistics version 19.0) and the Mann-Whitney rank-sum test.

[0123] 【2-3. Experimental Results】

[0124] (1) Weight measurement results

[0125] Table 2 and Figure 1 The results of body weight measurement on day 22 (dissection day) after administration of the test substance are shown.

[0126] Table 2

[0127]

[0128] Compared with G1 (solvent control group), the weight loss rates in all test substance administration groups (G3-G8) were 10.0%, 10.0%, 10.0%, 15.4%, 12.4%, and 17.4%, respectively, none exceeding 20%, and were therefore judged as not having reached the critical minimum weight. Figure 1 ).

[0129] (2) Tumor volume measurement results

[0130] Table 3 below and Figure 2 The results of volume measurements of pancreatic cancer tumors are shown in the figure.

[0131] Table 3

[0132]

[0133] In the solvent control group (G1) and the nab-paclitaxel monotherapy group (G2; 5 mg / kg), the volume of pancreatic cancer tumors continued to increase until day 22 after grouping, and the tumor growth rate was higher than that in other groups.

[0134] Compared with the solvent control group, the tumor volume of the nab-paclitaxel + SNB-101 (high dose) combination group (G8) decreased significantly from day 7. From day 17 to day 22, the tumor volume of both the SNB-101 (low and high dose) monotherapy groups (G5, G6) and the nab-paclitaxel + SNB-101 (high dose) combination group (G8) decreased significantly compared with the solvent control group (p<0.05).

[0135] On day 22, the group receiving the combination of nab-paclitaxel and irinotecan hydrochloride (G4) showed a 32.4% reduction compared to the nab-paclitaxel monotherapy group (G2) and a 12.8% reduction compared to the irinotecan hydrochloride monotherapy group (G3), but no statistically significant difference was observed (p>0.05).

[0136] When comparing the SNB-101 (low and high dose) monotherapy groups (G5 vs. G6), the high-dose monotherapy group (G6) showed a significant reduction of 11.0% compared to the low-dose monotherapy group (G5), indicating that the tumor-suppressive effect of SNB-101 in pancreatic cancer is dose-dependent (SN-38 / irinotecan hydrochloride at 10 / 15.9 mg / kg vs. 20 / 31.8 mg / kg).

[0137] On the other hand, when comparing the nab-paclitaxel monotherapy group (G2) or the SNB-101 (low and high doses) monotherapy groups (G5, G6) with the nab-paclitaxel + SNB-101 (low and high doses) combination therapy groups (G7, G8), both combination therapy groups showed stronger tumor growth inhibition effects than the individual monotherapy groups (p<0.05, p<0.01). When comparing the nab-paclitaxel + SNB-101 (low and high doses) groups (G7, G8), the tumor growth inhibition effect also increased statistically significantly with increasing SNB-101 dose (p<0.05).

[0138] On the other hand, no tumor-suppressive effect was confirmed in the nab-paclitaxel (5 mg / kg) monotherapy group (G2), but a stronger tumor growth inhibition effect (52.6%) was observed in the nab-paclitaxel + SNB-101 high-dose group (G8) (SN-38 / irinotecan hydrochloride 20 / 31.9 mg / kg) than in the SNB-101 high-dose monotherapy group (G6) alone (G6). Furthermore, when comparing the nab-paclitaxel + SNB-101 high-dose group (G8) with the nab-paclitaxel + irinotecan hydrochloride combination group (G4), a statistically significant and superior tumor growth inhibition effect was also observed (29.5%) (p<0.05).

[0139] The above results indicate that when nab-paclitaxel and SNB-101 are administered in combination, they exhibit a synergistic effect in inhibiting tumor growth.

[0140] (3) Tumor weight measurement results

[0141] Table 4 below and Figure 3 The results of tumor weight measurement are shown in the figure.

[0142] Table 4

[0143] G1 0.623±0.117 G2 0.636±0.150 G3 0.489±0.101 G4 0.440±0.096 G5 0.474±0.102 G6 0.379±0.112 G7 0.431±0.097 G8 0.310±0.050

[0144] As shown in Table 4 above and Figure 3As shown, the mean tumor weight was 0.62±0.11 g in the untreated group (solvent control group, G1), 0.64±0.15 g in the nab-paclitaxel treatment group (G2), and 0.47±0.10 g and 0.38±0.11 g in the SNB-101 (low and high dose) monotherapy groups (G5 and G6), respectively. Furthermore, the mean tumor weights in the nab-paclitaxel + SNB-101 (low and high dose) combination therapy groups (G7 and G8) were 0.43±0.10 g and 0.31±0.05 g, respectively. The combination therapy groups showed tumor inhibition effects of more than 50% compared to the nab-paclitaxel monotherapy group (G2) and more than 10–22.5% compared to the SNB-101 (low and high dose) monotherapy groups (G5 and G6). The above results confirm that nab-paclitaxel and SNB-101 have the potential to be a highly effective combination therapy for pancreatic cancer.

[0145] The combination of nab-paclitaxel and irinotecan hydrochloride (G4) significantly reduced the incidence by 30.8% compared to the nab-paclitaxel monotherapy group (G2). When comparing the SNB-101 (low and high dose) monotherapy groups, the high dose group (G6) reduced the incidence by 20.0% compared to the low dose group (G5).

[0146] [Example 3: The effect of combined drug delivery of the nanoparticles (SNB-101) of the present invention on gastric cancer]

[0147] 【3-1. Experimental Materials】

[0148] The substances used in the experiment included: 5% glucose injection and irinotecan hydrochloride (Calmtop Injection). TM CJ Healthcare, South Korea), Docetaxel (Taxotere Injection) TM Sanofi-Aventise Korea, and a nanoparticle composition comprising SN-38 and irinotecan hydrochloride (named SNB-101). The aforementioned irinotecan hydrochloride (Calmtop Injection) TMThe dosage form (100 mg / 5 mL) of docetaxel was purchased from CJ Healthcare. The dosage form (20 mg / mL) of docetaxel was purchased from Sanofi-Aventis Korea. The nanoparticle composition (SNB-101) containing the above-mentioned SN-38 and irinotecan hydrochloride used in this invention is an injectable preparation. It is a stable nanoparticle dosage form, and its preparation method is described in Korean Patent No. 10-2094543. Each vial of the above-mentioned SNB-101 intravenous injection contains 10 mg of SN-38 and 15.9 mg of irinotecan hydrochloride. It can be prepared by diluting with 9.64 mL of 5% glucose injection (USP standard), and the infusion time can be 90 minutes.

[0149] 【3-2. Test Methods】

[0150] (1) Culture of gastric cancer cell line Hs746T

[0151] Hs746T was used as the gastric cancer cell line for a gastric cancer xenograft model. [The text abruptly shifts to a seemingly unrelated topic:] ...at 175-cm... 2 Hs746T cells were cultured in flasks using Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum and 1% penicillin / streptomycin. Fetal bovine serum, DMEM, and penicillin-streptomycin were purchased from the American Culture Collection (Manassas, Virginia) and used in the experiments. Cells were cultured at 37°C, 95% air, and 5% CO2. Each cultured cancer cell was fed 2–3 times per week, washed with phosphate-buffered saline (pH 7.4), and then separated from the adherent cells using 0.05% trypsin-0.02% EDTA. After centrifugation (3 minutes, 1500 rpm), the culture medium was added to the accumulated cancer cells, and the cells were thoroughly mixed with a pipette until evenly dispersed. The cells were then used for subculture and experiments.

[0152] On the day of inoculation, after washing the cultured cancer cells with phosphate-buffered saline (PFS), adherent cells were separated using 0.05% trypsin-0.02% EDTA. The separated cells were centrifuged (3 minutes, 1500 rpm), diluted with PFS, and 4 × 10⁴ cells were prepared for each cell. 6 1 cell / 0.2 mL.

[0153] (2) Induction of a mouse model of gastric cancer xenograft

[0154] The animals used in this experiment were male BALB / c nu / nu mice (approximately 5 weeks old, with an average weight of 19g ± 20%), widely used in xenograft animal models, which underwent a acclimatization process of about 5-7 days after purchase. On the day of inoculation, the cultured cancer cells were washed with phosphate-buffered saline (pH 7.4), and then adherent cells were separated using 0.05% trypsin-0.02% EDTA. The separated cells were centrifuged (3 minutes, 1500 rpm), diluted with phosphate-buffered saline, and then injected at 5 × 10⁻⁶ cells per mouse. 6 One cell / 0.2 mL was subcutaneously injected into the right rib of the mouse to confirm that the tumor cell suspension did not flow out from the injection site.

[0155] [Group composition and dosage settings]

[0156] With 80±20mm 3 Mice were grouped based on the average gastric cancer tumor volume. The composition and dosage of the control and experimental groups are as follows:

[0157] Table 5

[0158]

[0159] * Dosage of SNB-101 labeled with SN-38 / irinotecan hydrochloride

[0160] (3) Administration site and administration method

[0161] The test substance was administered via tail vein after the individual's pre-administration weight was calculated. The test substance was administered once a week (on days 0, 7, and 14) for a total of 3 times, and dissection was performed on day 15.

[0162] (4) Observation and inspection items

[0163] Clinical symptoms were observed once daily during the experiment, and tumor volume was measured three times weekly while the drugs were being administered. At the end of the experiment, animals were anesthetized with isoflurane and bled, and tumor tissue was removed from each individual. The removed tumors were weighed and photographed. Tumor volume was measured using vernier calipers (Mitutoyo Corporation, model: CD-15CPX, Japan) along the major and minor axes, and calculated according to Equation 1 below.

[0164] Formula 1

[0165] Tumor volume = (short axis) 2 ×major axis × 0.5

[0166] (5) Statistical Analysis

[0167] The experimental results were statistically analyzed using a commercial statistical program (IBM SPSS statistics version 19.0) and the Mann-Whitney rank-sum test.

[0168] 【3-3. Experimental Results】

[0169] (1) Weight measurement results

[0170] Table 6 and Figure 5 The results of body weight measurement on day 15 (dissection day) after administration of the test substance are shown.

[0171] Table 6

[0172] G1 21.2±1.2 21.6±1.0 21.7±1.1 21.7±1.1 22.7±1.3 22.2±1.1 G2 21.9±1.6 22.5±1.6 22.4±1.3 22.3±1.5 22.8±1.4 22.2±1.4 G3 21.1±1.3 21.7±1.1 21.7±1.1 21.8±1.1 22.8±1.1 21.9±1.1 G4 22.0±0.9 21.4±0.7 21.7±0.7 20.8±0.8 21.6±1.2 20.1±1.1 G5 21.5±1.2 21.8±1.0 21.9±0.8 21.8±1.3 23.1±1.5 21.5±1.3 G6 21.0±1.6 20.3±2.5 20.6±2.6 19.1±1.6 20.7±1.9 19.1±1.9

[0173] Unit: g; data are expressed as mean ± standard deviation (n = 9).

[0174] G1: Solvent control group (Vehicle control) (5% glucose injection)

[0175] G2: Positive control group 1 (5 mg / kg / day docetaxel)

[0176] G3: Positive control group 2 (60 mg / kg / day irinotecan hydrochloride)

[0177] G4: Positive control group 3 (5 mg / kg / day docetaxel + 60 mg / kg / day irinotecan hydrochloride)

[0178] G5: Test substance 1 (20 / 31.8 mg / kg / day (SN-38 / irinotecan hydrochloride) SNB-101)

[0179] G6: Test substance 2 (5 mg / kg / day docetaxel + 20 / 31.8 mg / kg / day (SN-38 / irinotecan hydrochloride) SNB-101)

[0180] No significant differences in body weight were observed in any group at the time of purchase and grouping. Body weight was measured on day 15 (dissection day) of test substance administration. Weight loss was observed in the G4 (docetaxel + irinotecan hydrochloride) and G6 (docetaxel + SNB-101) combination groups. On day 15, the weight of the combination groups was significantly lower than that of the excipient control groups (G1 (5% glucose injection), G2 (docetaxel), G3 (irinotecan hydrochloride), and G4 (SNB-101) single-drug groups). Compared to G1 (solvent control group), the weight loss rate in the test substance groups (G2-G6) did not exceed 20%, and was therefore considered not to have reached the critical minimum body weight. Figure 5 ).

[0181] (2) Tumor volume measurement results

[0182] Table 7 below and Figure 6 The figure shows the volume of the gastric cancer tumor (mm). 3 The measurement results.

[0183] Table 7

[0184]

[0185] Unit: g, data expressed as mean ± standard deviation (n=9)

[0186] *Compared with G1:*p<0.05,**p<0.01; # Compared with G1: # p<0.05, ## p<0.01; Compared with G3:

[0187] G1: Solvent control group (5% glucose injection)

[0188] G2: Positive control group 1 (5 mg / kg / day docetaxel)

[0189] G3: Positive control group 2 (60 mg / kg / day irinotecan hydrochloride)

[0190] G4: Positive control group 3 (5 mg / kg / day docetaxel + 60 mg / kg / day irinotecan hydrochloride)

[0191] G5: Test substance 1 (20 / 31.8 mg / kg / day (SN-38 / irinotecan hydrochloride) SNB-101)

[0192] G6: Test substance 2 (5 mg / kg / day docetaxel + 20 / 31.8 mg / kg / day (SN-38 / irinotecan hydrochloride) SNB-101)

[0193] In the measurement of gastric cancer tumor volume, the excipient control group G1 continued to increase until day 15 after grouping, and the tumor growth rate was higher than that of other groups.

[0194] Compared with the excipient control group G1, the tumor volume of G4 (docetaxel + irinotecan hydrochloride combination group), G5 (SNB-101 alone group) and G6 (docetaxel + SNB-101 combination group) decreased significantly from day 7. On day 15, the tumor volume of the positive control group (G2, G3, G4) and the test substance group (G5, G6) was significantly reduced.

[0195] Starting from day 11, G4 (docetaxel + irinotecan hydrochloride combination group) was significantly reduced compared to G2 (docetaxel alone group). Starting from day 9, G6 (docetaxel + SNB-101 combination group) was significantly reduced compared to G2 (docetaxel alone group) and G3 (irinotecan hydrochloride alone group).

[0196] (3) Tumor weight measurement results

[0197] Table 8 below and Figure 7 The results of tumor weight measurement are shown in the figure.

[0198] Table 8

[0199]

[0200] Data are expressed as mean ± standard deviation (n = 9);

[0201] *Compared to G1: ** p<0.01;

[0202] # Compared to G2: # p<0.05, ## p<0.01;

[0203] Compared to G3:

[0204] G1: Solvent control group (5% glucose injection)

[0205] G2: Positive control group 1 (5 mg / kg / day docetaxel)

[0206] G3: Positive control group 2 (60 mg / kg / day irinotecan hydrochloride)

[0207] G4: Positive control group 3 (5 mg / kg / day docetaxel + 60 mg / kg / day irinotecan hydrochloride)

[0208] G5: Test substance 1 (20 / 31.8 mg / kg / day (SN-38 / irinotecan hydrochloride) SNB-101)

[0209] G6: Test substance 2 (5 mg / kg / day docetaxel + 20 / 31.8 mg / kg / day (SN-38 / irinotecan hydrochloride) SNB-101)

[0210] As shown in Table 8 above Figure 7 As shown, the results of tumor resection and tumor weight comparison showed significant reductions in both positive control substances (G2, G3, G4) and the single-drug and combined-drug groups (G5 and G6) compared to the excipient control group G1. Compared to G2 (docetaxel alone), significant reductions were also observed in G4 (docetaxel + irinotecan hydrochloride combination), G5 (SNB-101 alone), and G6 (docetaxel + SNB-101 combination). Furthermore, compared to the irinotecan hydrochloride alone group, significant reductions were observed in both the SNB-101 alone and docetaxel + SNB-101 combination groups.

[0211] Therefore, under the conditions of this experiment, in the xenograft model using the gastric cancer cell line Hs746T, the following substances were determined to have inhibitory effects on tumor growth: positive control substance G2 (5 mg / kg / day docetaxel), G3 (60 mg / kg / day irinotecan hydrochloride alone), G4 (5 mg / kg / day docetaxel + 60 mg / kg / day irinotecan hydrochloride combination), test substance G5 (20 / 31.8 mg / kg / day (with SN-38 / irinotecan hydrochloride) SNB-101 alone), and G6 (5 mg / kg / day docetaxel + 20 / 31.8 mg / kg / day (with SN-38 / irinotecan hydrochloride) SNB-101 combination). Compared with the positive control substance alone groups (G2, G3), the combination groups (G4, G6) showed stronger tumor inhibition effects.

[0212] [Example 4: The effect of combined drug delivery of the nanoparticles (SNB-101) of the present invention on breast cancer and lung cancer]

[0213] 【4-1. Experimental Materials】

[0214] The substances used in the experiment included: 5% glucose injection and docetaxel (Taxotere Injection). TM The drug was obtained from Sanofi-Aventis Korea, and a nanoparticle composition containing SN-38 and irinotecan hydrochloride (named SNB-101). Docetaxel was available at 20 mg / ml from Sanofi-Aventis Korea.

[0215] The nanoparticle composition (SNB-101) containing SN-38 and irinotecan hydrochloride used in this invention is an injectable preparation. It is a stable nanoparticle dosage form, and its preparation method is described in Korean Patent No. 10-2094543. Each vial of the above-mentioned SNB-101 intravenous injection contains 10 mg of SN-38 and 15.9 mg of irinotecan hydrochloride. It can be prepared by diluting with 9.64 mL of 5% glucose injection (United States Pharmacopeia standard), and the infusion time can be 90 minutes.

[0216] 【4-2. Test Methods】

[0217] (1) Culture of breast cancer cell line (MDA-MB-231) and lung cancer cell line (A549).

[0218] MDA-MB-231 and A549 were used as breast cancer and lung cancer cell lines for xenograft models. Frozen cell lines MDA-MB-231 and A549 were thawed and cultured in RPMI 1640 medium-10% fetal bovine serum-1% penicillin-streptomycin and RPMI 1640 medium-5% fetal bovine serum-1% penicillin-streptomycin, respectively, at 37°C and 5% CO2. Thawed cells were cultured for at least one week, with subculture every 2–3 days to confirm viability (trypan blue), and to confirm the absence of bacteria, yeast (cell images), and mycoplasma infection (MycoAlert Mycoplasma detection kit). Cells with a viability of over 90% were used for transplantation.

[0219] (2) Induction of mouse models of breast and lung cancer xenograft

[0220] The animals used in this experiment were male BALB / c nu / nu mice (approximately 5 weeks old, with an average weight of 19g ± 20%), widely used in xenograft animal models, which underwent a acclimatization process of about 5-7 days after purchase. On the day of inoculation, the cultured cancer cells were washed with phosphate-buffered saline (PBS, pH 7.4), and then adherent cells were separated using 0.05% trypsin-0.02% EDTA. The separated cells were centrifuged (3 minutes, 1500 rpm), diluted with phosphate-buffered saline, and MDA-MB-231 and A549 cells were injected at 5 × 10⁶ cells per mouse. 6 One cell / 100μL was subcutaneously transplanted into the right shoulder of Balb / c-nu mice to confirm that the tumor cell suspension did not leak out from the injection site.

[0221] (3) Group composition, dosage, administration site and administration method

[0222] When the average tumor volume of all mice was 90–105 mm 3 Mice were grouped using a paired-matching method. The composition and dosage of the control and experimental groups are shown in Tables 9 and 10.

[0223] Table 9: MDA-MB-231 Breast Cancer Cell Line

[0224]

[0225] *The dosage of docetaxel was changed from 5 mg / kg (dose 1-3) to 15 mg / kg (dose 4-6) during the trial.

[0226] **The frequency of drug administration in all groups was changed from once a week for 3 weeks (Q1W×3 times) to once a week for 6 weeks (Q1W×6 times) during the trial.

[0227] The dosage of 20 mg / kg for SNB-101 is based on the content of SN-38.

[0228] Table 10: Lung Cancer A549 Cell Line

[0229]

[0230]

[0231] *The dosage of docetaxel was changed from 5 mg / kg (first dose) to 10 mg / kg (second-fifth doses) during the trial.

[0232] **The frequency of drug administration in all groups was changed from once a week for 3 weeks (Q1W×3 times) to once a week for 5 weeks (Q1W×5 times) during the trial.

[0233] The dosage of 20 mg / kg for SNB-101 is based on the content of SN-38.

[0234] From the start of the grouping process until the end of the trial, tumor volume and body weight were measured twice a week (with an interval of 3 or 4 days), and tumor growth curves were plotted based on the tumor volume from the start of drug administration to the end of the trial.

[0235] When severe necrosis and weight loss are detected, in accordance with the regulations of the International Association for Animal Research and Ethics (IACUC), euthanasia is performed in consultation with the commissioning company to end the experiment, the tumor is removed, and photographs are taken.

[0236] [4-3. Trial Results of the MDA-MB-231 Xenograft Model for Breast Cancer]

[0237] (1) Measure tumor volume and analyze results

[0238] Using the end date of the experiment (day 35 after drug administration) as the baseline, the tumor growth inhibition efficacy of G2 (docetaxel alone), G3 (SNB-101 alone), and G4 (SNB-101 + docetaxel combination) was confirmed by comparison with G1 (solvent control group).

[0239] The mean tumor volume in the G1 (solvent), G2 (docetaxel), G3 (SNB-101), and G4 (SNB-101 + docetaxel) treatment groups was 1662.48 mm. 3 1228.93mm 3 657.30mm 3 364.10mm 3 The combined administration (G4) was more effective in inhibiting tumor formation than the single administration (G2, G3). The results are as follows: Figure 9 As shown.

[0240] The tumor growth inhibition rate (TGI) relative to the solvent control group, based on the experimental end date, is calculated using the following formula.

[0241] Formula 2

[0242] Tumor growth inhibition rate (%) = {1 - (ΔT / ΔC)} × 100

[0243] The tumor growth inhibition rates of each group are shown in Table 11.

[0244] Table 11

[0245] G2 (docetaxel) 27.6 G3 (SNB-101) 64.0 G4 (SNB-101 + Docetaxel) 82.6

[0246] As shown in Table 11, the tumor growth inhibition rate was 27.6% in the docetaxel monotherapy group (G2) compared to the solvent control group, 64.0% in the SNB-101 monotherapy group (G3), and 82.6% in the SNB-101 + docetaxel combination therapy group (G4). Based on these results, it can be confirmed that the combined administration of SNB-101 and docetaxel of the present invention exhibits a synergistic effect on breast cancer cells.

[0247] (2) Weight measurement results

[0248] The weight measurement results for each group are as follows: Figure 10 As shown in the image.

[0249] At the start of administration (day 0), the body weights of the solvent control group, the docetaxel monotherapy group, the SNB-101 monotherapy group, and the SNB-101 + docetaxel combination group were 19.70 g, 19.63 g, 20.99 g, and 21.46 g, respectively. At the end of the trial (day 35), the body weights were 22.46 g, 20.25 g, 22.43 g, and 19.37 g, respectively. This confirms that the body weight loss rate did not exceed 20%, and the critical minimum body weight was not reached.

[0250] (3) Conclusion

[0251] In summary, in the MDA-MB-231 xenograft model of breast cancer cell line, the combination of SNB-101 and docetaxel (G4) showed superior tumor growth inhibition compared to the administration of docetaxel (G2) and SNB-101 alone (G3).

[0252] [4-4. Results of the A549 xenograft model trial for lung cancer]

[0253] (1) Tumor volume measurement and result analysis

[0254] Using the end date of the experiment (day 37 after drug administration) as the baseline, the tumor growth inhibition efficacy of G2 (docetaxel alone), G3 (SNB-101 alone), and G4 (SNB-101 + docetaxel combination) was confirmed by comparison with G1 (solvent control group).

[0255] The mean tumor volume in the G1 (solvent), G2 (docetaxel), G3 (SNB-101), and G4 (SNB-101 + docetaxel) treatment groups was 704.10 mm. 3 402.70mm 3557.60mm 3 181.08mm 3 The combination therapy (G4) was more effective in inhibiting tumor formation compared to the single therapy (G2, G3). The results are as follows: Figure 11 As shown.

[0256] The tumor growth inhibition rate relative to the solvent control group, based on the end date of the experiment, is obtained using the following formula.

[0257] Formula 2

[0258] Tumor growth inhibition rate (%) = {1 - (ΔT / ΔC)} × 100

[0259] The tumor growth inhibition rates of each group are shown in Table 12.

[0260] Table 12

[0261] G2 (docetaxel) 50.0 G3 (SNB-101) 24.3 G4 (SNB-101 + Docetaxel) 86.7

[0262] As shown in Table 12, the tumor growth inhibition rate was 50.0% in the docetaxel monotherapy group (G2) compared to the solvent control group, 24.3% in the SNB-101 monotherapy group (G3), and 82.6% in the SNB-101 + docetaxel combination group (G4).

[0263] Based on the above results, it can be confirmed that when the SNB-101 of the present invention and docetaxel are administered in combination, a synergistic effect is observed on the lung cancer cell line A549.

[0264] (2) Weight measurement results

[0265] The weight measurement results for each group are as follows: Figure 12 As shown in the image.

[0266] At the start of administration (day 0), the body weights of the solvent control group, the docetaxel monotherapy group, the SNB-101 monotherapy group, and the SNB-101 + docetaxel combination group were 17.86g, 17.93g, 18.04g, and 18.68g, respectively. At the end of the trial (day 37), the body weights were 20.19g, 19.10g, 19.79g, and 17.97g, respectively. That is, the confirmed weight loss rate did not exceed 20%, and the critical minimum body weight was not reached.

[0267] (3) Conclusion

[0268] In summary, in the A549 lung cancer cell line xenograft model, the combination of SNB-101 and docetaxel (G4) showed superior tumor growth inhibition compared to the administration of docetaxel (G2) and SNB-101 alone (G3).

[0269] [Example 5: Evaluation of the synergistic effect of combined administration of the present invention's nanoparticles (SNB-101) and bevacizumab (trade name: Avastin) on the treatment of colorectal cancer]

[0270] An experiment was conducted to evaluate the synergistic effect of combination therapy with bevacizumab, a monoclonal antibody and targeted anticancer agent, in the treatment of colorectal cancer.

[0271] 【5-1. Experimental Methods】

[0272] (1) Laboratory animals

[0273] A total of 150 male BALB / c nude mice (5 weeks old) were purchased and acclimatized for about one week. HT-29 colorectal cancer cells were transplanted into the subcutaneous part of the right buttock. Seven days after the tumor cells were transplanted, 50 mice were selected again for transplantation based on the tumor volume, with 10 mice in each group, for this experiment.

[0274] (2) Transplantation of tumor cells

[0275] HT-29 (KCLB No. 300038, Korea Cell Bank) human colorectal cancer cells were subcultured and maintained in an incubator at 37°C and 5% CO2 using RPMI 1640 medium supplemented with 10% fetal bovine serum and 100 IU / ml penicillin / streptomycin to prepare a concentration of 3.0 × 10⁻⁶ cells / mL. 6 One cell per 0.1 ml of tumor cell suspension was used to transplant 0.1 mL of HT-29 tumor cells into the subcutaneous area of ​​the right buttock of each mouse, forming a solid mass.

[0276] (3) Grouping

[0277] Eight days after transplantation of HT-29 colorectal cancer cells, mice were selected based on tumor volume and randomly divided into 5 groups (n=10 per group). The drug administration groups are shown in Table 13 below.

[0278] Table 13

[0279] G1 Solvent (5% glucose) 10 - vein G2 SNB-101 10 20 / 31.8* vein G3 bevacizumab 10 5 abdominal cavity G4 Irinotecan + bevacizumab 10 50+5 vein + abdominal cavity G5 SNB-101 + Bevacizumab 10 20 / 31.8*+5 vein + abdominal cavity

[0280] * Dosage of SN-38 / irinotecan hydrochloride

[0281] (4) Drug administration

[0282] The test substance SNB-101 and bevacizumab (5 mg / kg) were administered three times on the same day (days 1, 4, and 7), either alone or in combination. Dosage was calculated for each individual based on body weight at administration. SNB-101 was administered intravenously, and bevacizumab was administered intraperitoneally. Dissection was performed on day 28. As a control group, a 5% glucose solution (used as a vehicle) was administered at a dose of 10 mL / kg.

[0283] 【5-2. Experimental Results】

[0284] The test results are shown in Table 14 below as mean ± standard deviation.

[0285] Table 14

[0286]

[0287] As shown in Table 14 above, the combination therapy group (G5) was found to have a superior synergistic effect in the treatment of colorectal cancer compared with the SNB-101 monotherapy group (G2) and the bevacizumab monotherapy group (G3). Figure 13 The tumor growth curves corresponding to the data in Table 14 above are shown. The statistical significance of differences in tumor volume between groups at different time points was analyzed using the Holm-Sidak multiple comparison test and two-way ANOVA. * P<0.05; ** P<0.01; *** P < 0.001, ns: non-significant.

[0288] [Example 6: Evaluation of the synergistic effect of combined administration of the present invention's nanoparticles (SNB-101), nab-paclitaxel, and gemcitabine on the treatment of pancreatic cancer]

[0289] 【6-1. Experimental Materials】

[0290] BALB / c-nu mice (ORIENT BIO Gapyeong Center), male, 6 weeks old

[0291] Pancreatic cancer AsPC-1 cell line (American Culture Collection, Catalogue No. CRL-1682)

[0292] Investigational drugs: SNB-101 (SN-38 10mg, irinotecan hydrochloride·3H2O 15.9mg / vial), nab-paclitaxel (Celgene, South Korea, Abraxane, Lot No. 6200676A), gemcitabine (Lilly, South Korea, Lot No. 186053A)

[0293] Solvent: 50ml of 5% glucose injection (CJ Healthcare Company, Insurance Code 640001361)

[0294] RPMI 1640 medium (Gibco, A10491-01) supplemented with 10% fetal bovine serum (Gibco, Catalogue No. 16000)

[0295] 1% Penicillin-Streptomycin (Gibco, Catalog No. 15140122)

[0296] Trypsin-EDTA (Gibco, catalog number 25200-072)

[0297] Cold-Duborough Phosphate Buffered Solution (DPBS) (Hyclone, catalog number SH30258.02)

[0298] Trypan blue (Gibco, catalog number 15250061)

[0299] MycoAlert Mycoplasma Detection Kit (LONZA, Catalog No. LT-07-318)

[0300] 150mm cell culture dish, conical tubes, and pipettes.

[0301] Mouse ear tag, syringe, calipers

[0302] 【6-2. Test Methods】

[0303] Pancreatic cancer cell line AsPC-1, provided by the National Center for the Establishment of T2B Infrastructure for Digestive Diseases (NCEED), was cultured in RPMI 1640 medium-10% fetal bovine serum-1% penicillin-streptomycin at 37°C and 5% CO2. Thawed cells were cultured for at least one week, subcultured every 2-3 days to confirm viability and to ensure the absence of cell, yeast, and MycoAlert mycoplasma infection (MycoAlert Mycoplasma detection kit). Cells with a viability of over 90% were used for transplantation after trypsin-EDTA suspension. AsPC-1 cells were cultured at 5 × 10⁶ cells / day. 6 200 μl cells / cell were subcutaneously transplanted into the right thigh of BALB / c-nu mice. Tumor formation and growth were observed periodically, and tumor volume was calculated using the measured tumor length according to the following formula.

[0304] [Tumor volume = (a 2 b) / 2, where a is the shorter diameter and b is the longer diameter.

[0305] On the day of administration, the average tumor volume of all mice was 123.6 mm. 3 Mice were grouped using a paired-matching method. Following administration to the mice according to the dosages and methods described in Table 15, the mice were observed for two weeks.

[0306] Table 15

[0307]

[0308]

[0309] * Dosage of SN-38 / irinotecan hydrochloride

[0310] From the start of the grouping process until the end of the trial, tumor volume and body weight were measured twice a week (every 3 or 4 days apart), and the degree of inhibition of tumor growth from the start of drug administration to the end of the trial was evaluated.

[0311] 【6-3. Experimental Results】

[0312] Using Equation 2 above, the tumor growth inhibition rate (TGI) relative to the solvent control group was obtained as of the end date of the experiment.

[0313] The tumor growth inhibition rates of each group are shown in Table 16.

[0314] Table 16

[0315]

[0316] As shown in Table 16 above, the tumor growth inhibition rate was 72.5% in the SNB-101 monotherapy group (G2) compared to the solvent control group, and 56.3% in the nab-paclitaxel + gemcitabine group (G3). Conversely, the tumor growth inhibition rate was 83.4% in the SNB-101 + nab-paclitaxel + gemcitabine combination group (G4), confirming that the combination of SNB-101 + nab-paclitaxel + gemcitabine has a very excellent synergistic therapeutic effect in the treatment of pancreatic cancer.

[0317] Meanwhile, no side effects, weight loss, or toxicity were observed in blood analyses in any of the treatment groups.

[0318] [Example 7: Evaluation of the synergistic effect of combined use of the present invention's nanoparticle (SNB-101) drug delivery and radiation irradiation in the treatment of colorectal cancer and lung cancer]

[0319] 【7-1. Experimental Materials】

[0320] BALB / c-nu mice (JungA Biotechnology), male, 6 weeks old

[0321] Small cell lung cancer NCI-H69 cell line (USCSC, catalog number HTB-119)

[0322] Non-small cell lung cancer A549 cell line (American Culture Collection, catalog number CCL-185)

[0323] HCT116 cell line for colorectal cancer (USCSC, catalog number CCL-247)

[0324] Investigational drug: SNB-101 (SN Bioscience Co., Ltd., SN-38 10mg, irinotecan hydrochloride·3H2O 15.9mg / vial)

[0325] Solvent: 50ml of 5% glucose injection (CJ Healthcare Company, Insurance Code 640001361)

[0326] RPMI 1640 medium (Gibco, 22400-089), F12K medium (Gibco, 21127-022), and McCoy's 5A medium (Gibco, 16600-082) supplemented with 10% fetal bovine serum (Gibco, catalog number 16000)

[0327] 1% Antibiotic-Antifungal Agent (Gibco, Catalog No. 15240-062)

[0328] Trypsin-EDTA (Gibco, catalog number 25200-072)

[0329] Cold-Duborough phosphate buffer (Gibco, catalog number 14200-075)

[0330] Trypan blue (Gibco, catalog number 15250061)

[0331] MycoAlert Mycoplasma Detection Kit (LONZA, Catalog No. LT-07-318)

[0332] 150mm cell culture dish, conical tube, pipette

[0333] Mouse ear tags, syringes, calipers

[0334] 【7-2. Test Methods】

[0335] Frozen cell lines NCI-H69 (small cell lung cancer), HCT116 (colorectal cancer), and A549 (non-small cell lung cancer) from the Anti-Cancer T2B Infrastructure Construction Center were thawed and cultured at 37°C and 5% CO2 using RPMI 1640, F12K, and McCoy's 5A medium-10% fetal bovine serum-1% antibiotic-antifungal agent, respectively. Thawed cells were cultured for at least one week, with subculture every 2-3 days to confirm viability and to ensure the absence of cell, yeast, and MycoAlert mycoplasma infection (MycoAlert Mycoplasma detection kit). Trypan blue staining was used, and cells with a viability of over 90% were used for transplantation. Cells were suspended in trypsin-EDTA and then resuspended in cold phosphate buffer. NCI-H69 cells were cultured at 1×10⁻⁶ cells / mL. 7 One cell / 100 μl, and A549, HCT116 cells at 1×10 6 50 μl of cells were subcutaneously transplanted into the right thigh of BALB / c-nu mice. Tumor formation and growth were observed periodically, and the tumor volume was calculated using the measured tumor length according to the following formula.

[0336] [Tumor volume = (a 2 b) / 2, where a is the shorter diameter and b is the longer diameter.

[0337] On the day of administration, the average tumor volume of all mice was 100 ± 20 mm. 3Mice were grouped using a paired method. Following administration of the drugs to mice according to the dosages and administration methods described in Tables 17 and 15, mice were observed for 2-3 weeks.

[0338] The NCI-H69 and HCT116 models were administered the drug once a week for a total of one dose, while the A549 model was administered the drug twice a week for a total of five doses. After administration, tumor volume and body weight were measured in the HCT116 model until day 21, in the NCI-H69 model until day 38, and in the A549 model until day 45.

[0339] [Universal xenograft model for NCI-H69 (small cell lung cancer) and HCT116 (colorectal cancer)]

[0340] Table 17

[0341]

[0342] * Dosage of SN-38 / irinotecan hydrochloride

[0343] [A549 (Non-Small Cell Lung Cancer) Xenograft Model (General)]

[0344] Table 18

[0345]

[0346] * Dosage of SN-38 / irinotecan hydrochloride

[0347] ** In the A549 xenograft model, the dosage of SNB-101 was changed through mutual consultation during the experiment from 20 mg / kg (first time) → 30 mg / kg (second time, after the death of an individual) → 20 mg / kg (third-fifth times).

[0348] *** The dosing frequency in the SNB-101 group was changed from 9 days after the first dose to 4 doses of BIW for a total of 2 weeks (5 doses).

[0349] From the start of the grouping process until the end of the experiment, tumor volume and body weight were measured twice a week (every 3 or 4 days apart), and tumor growth curves were plotted based on tumor volume from the start of drug administration to the end of the experiment. When the tumor volume reached the limit stipulated by the International Association for Animal Ethics (IACUC), euthanasia was performed in consultation with the commissioning company, the experiment was terminated, and the tumor was removed for analysis.

[0350] 【7-3. Experimental Results】

[0351] Using Equation 2 above, the tumor growth inhibition rate relative to the solvent control group was obtained, with the end date of the experiment as the baseline.

[0352] In the case of NCI-H69, the tumor growth inhibition rates of each group are shown in Table 19.

[0353] Tumor volume change curve at Figure 14 As shown in the image.

[0354] Tumor resection photos in Figure 15 As shown in the image.

[0355] Table 19

[0356] G2 SNB-101 5 20 / 31.8mg / kg 50.9 G3 RT 5 5Gy 81.3 G4 SNB-101+RT 5 20 / 31.8Mg / kg + 5Gy 101.5

[0357] In the case of HCT116, the tumor growth inhibition rates of each group are shown in Table 20.

[0358] Tumor volume change curve at Figure 16 As shown in the image.

[0359] Tumor resection photos in Figure 17 As shown in the image.

[0360] Table 20

[0361] G2 SNB-101 5 20 / 31.8mg / kg 62.6 G3 RT 5 5Gy 41.4 G4 SNB-101+RT 5 20 / 31.8 mg / kg + 5 Gy 95.4

[0362] In the case of A549, the tumor growth inhibition rates of each group are shown in Table 21.

[0363] Tumor volume change curve at Figure 18 As shown in the image.

[0364] Table 21

[0365] G2 SNB-101 5 20 / 31.8mg / kg -11.2 G3 RT 5 5Gy 33.9 G4 SNB-101+RT 5 20 / 31.8Mg / kg + 5Gy 49.0

[0366] As shown in Tables 19-21, the tumor growth inhibition rate relative to the solvent control group was significantly higher in the combined treatment group (G4) compared to the SNB-101 monotherapy group (G2) and the radiation monotherapy group (G3). This confirms that the combined treatment of SNB-101 and radiation therapy has a very excellent synergistic effect in the treatment of colorectal cancer and lung cancer.

[0367] Meanwhile, no side effects, weight loss, or toxicity were observed in blood analyses in any of the treatment groups.

[0368] In summary, through the aforementioned Examples 2 to 7, it was confirmed that the combined drug administration or combined radiotherapy as summarized in Table 22 achieved a significant synergistic effect in the treatment of cancer.

[0369] Table 22

[0370]

[0371]

Claims

1. Use of a pharmaceutical combination in the preparation of a medicament for the treatment of pancreatic cancer, wherein the combination comprises the following components as an active ingredient: Particles containing SN-38 (7-ethyl-10-hydroxycamptothecin), irinotecan, and amphiphilic block copolymers composed of hydrophobic and hydrophilic blocks; and Antitumor agents, namely albumin-bound nanoparticle paclitaxel or a combination of albumin-bound nanoparticle paclitaxel and gemcitabine. The amphiphilic block copolymer is composed of AB blocks or ABA blocks. Wherein (a) A is a hydrophilic polymer, and is monomethoxy polyethylene glycol, dimethoxy polyethylene glycol, polyethylene glycol, polypropylene glycol, monomethoxy polyethylene glycol, polyethylene oxide, or polyacrylic acid; and (b) B is a hydrophobic polymer, which is poly-L-lactide, poly-D-lactide, poly-D,L-lactide, poly(lactide-co-glycolic acid), polyglycolic acid, polylactic acid-gluconic acid copolymer, polymandelic acid, polycaprolactone, polydioxane-2-one, polyglutamic acid, polyaspartic acid, polyornithine, polyorthoester or its derivatives.

2. Use of a combination of pharmaceutical preparations in the preparation of a medicament for the treatment of gastric cancer, breast cancer, or lung cancer, wherein said combination of preparations comprises the following components as an active ingredient: Particles containing SN-38 (7-ethyl-10-hydroxycamptothecin), irinotecan, and amphiphilic block copolymers composed of hydrophobic and hydrophilic blocks; and Docetaxel, The amphiphilic block copolymer is composed of AB blocks or ABA blocks. Wherein (a) A is a hydrophilic polymer, and is monomethoxy polyethylene glycol, dimethoxy polyethylene glycol, polyethylene glycol, polypropylene glycol, monomethoxy polyethylene glycol, polyethylene oxide, or polyacrylic acid; and (b) B is a hydrophobic polymer, which is poly-L-lactide, poly-D-lactide, poly-D,L-lactide, poly(lactide-co-glycolic acid), polyglycolic acid, polylactic acid-gluconic acid copolymer, polymandelic acid, polycaprolactone, polydioxane-2-one, polyglutamic acid, polyaspartic acid, polyornithine, polyorthoester or its derivatives.

3. Use of a combination drug formulation in the preparation of a medicament for the treatment of colorectal cancer, wherein the combination drug formulation comprises the following components as an active ingredient: Particles containing SN-38 (7-ethyl-10-hydroxycamptothecin), irinotecan, and amphiphilic block copolymers composed of hydrophobic and hydrophilic blocks; and Bevacizumab, The amphiphilic block copolymer is composed of AB blocks or ABA blocks. Wherein (a) A is a hydrophilic polymer, and is monomethoxy polyethylene glycol, dimethoxy polyethylene glycol, polyethylene glycol, polypropylene glycol, monomethoxy polyethylene glycol, polyethylene oxide, or polyacrylic acid; and (b) B is a hydrophobic polymer, which is poly-L-lactide, poly-D-lactide, poly-D,L-lactide, poly(lactide-co-glycolic acid), polyglycolic acid, polylactic acid-gluconic acid copolymer, polymandelic acid, polycaprolactone, polydioxane-2-one, polyglutamic acid, polyaspartic acid, polyornithine, polyorthoester or its derivatives.

4. The use according to any one of claims 1 to 3, wherein the average diameter of the particles is 2 to 200 nm.

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