Nano-drug delivery platform loaded with ultra-small anticancer drug having multiple mechanisms of action, pharmaceutical composition containing same, and preparation method therefor

Abstract

The present invention relates to a nano-drug delivery platform loaded with an ultra-small anticancer drug having multiple mechanisms of action, a pharmaceutical composition containing same, and a preparation method therefor. The present invention can provide a drug carrier-based ultra-small anticancer agent, which enables treatment even at dosages lower than that of complete anticancer drugs used in a conventional clinical trial, and thus has no effect on normal tissue and achieves high patient compliance, and can reduce side effects during or after treatment.

Description

Nano drug delivery platform loaded with ultrafine anticancer drugs having multiple mechanisms of action, pharmaceutical composition comprising the same, and method for manufacturing the same

[0001] The present invention relates to a nano drug delivery platform loaded with an ultrafine anticancer drug having multiple mechanisms of action, a pharmaceutical composition comprising the same, and a method for producing the same.

[0002] Cancer is primarily treated through one or a combination of surgical resection, radiation therapy, and chemotherapy. Surgical resection involves direct removal of the tumor tissue through wide excision. Therefore, while effective, its use is limited. In cases of leukemia, diffuse tumors, or those that are difficult to treat with surgery, chemotherapy is used to treat the tumor. Systemic side effects (arthralgia, muscle pain, loss of appetite, fever, nausea, vomiting, diarrhea, indigestion, hair loss, and decreased white blood cells (neutrophils)) that occur during treatment, including the most common side effects of chemotherapy, motion sickness and vomiting, have the greatest impact on patients undergoing cancer treatment. They can negatively impact their quality of life and drastically alter treatment compliance. Furthermore, chemotherapy typically has dose-limiting toxicities (DLTs), which limit the dose administered due to solubilizing agents or their side effects. Even with limited doses, severe side effects may require hospitalization or painkillers to treat the pain.

[0003] Existing anticancer drugs are often administered in liquid form, typically formulated with surfactants and ethanol as solubilizers, to enhance their efficacy. For convenience, these drugs are often administered in pill form, typically in the form of small molecules simply mixed with various excipients. These cytotoxic finished anticancer drugs, however, inevitably act systemically, indiscriminately affecting both normal and cancer cells, resulting in numerous side effects. Therefore, research is actively underway to develop new anticancer drugs with improved functionality and lower toxicity, utilizing novel mechanisms that minimize the impact on normal cells and enhance their action on cancer cells.

[0004] The well-known 'Taxol' ® Paclitaxel (anticancer drug) is a chemical anticancer drug that inhibits cancer cell proliferation by interfering with the separation of microtubules, the machinery of cell division and self-replication, and has played a crucial role in the treatment of cancer patients. Paclitaxel, a natural substance extracted from the bark of the yew tree, has garnered significant attention and is considered one of the most important anticancer drugs used over the past decade.

[0005] Taxane drugs, including docetaxel and paclitaxel, are used as first-line therapy for metastatic cancers, including breast cancer, ovarian cancer, and prostate cancer. Recently, their effectiveness has been confirmed in combination with targeted or immunotherapy drugs, and their scope of use is expanding.

[0006] Paclitaxel, the most widely used taxane-based anticancer drug worldwide, has been clinically studied as a preoperative neoadjuvant chemotherapy, postoperative adjuvant therapy, and second-line therapy, and is currently being used in clinical trials in various combination studies with immunotherapy drugs, so its range of applications is very wide.

[0007] Most cancer patients previously treated with taxane-based anticancer drugs have reportedly developed resistance to these chemotherapy regimens, leaving several challenges to overcome. Paclitaxel, a first-line anticancer drug used in the treatment of ovarian, breast, and lung cancers, is "insoluble in water, so it tends to precipitate when mixed with body fluids." Furthermore, it is considered an anticancer drug with low bioavailability even after absorption, making it difficult to commercialize orally. Many pharmaceutical companies worldwide have failed to develop paclitaxel. However, domestic pharmaceutical companies are actively participating in the development and commercialization of oral anticancer drugs, driven by their numerous advantages, demand, and economic value, alongside technological advancements. Successful development of platform-based drug delivery technologies is expected to increase cancer cell sensitivity to these drugs, contributing to the development of novel targeted drugs with high therapeutic efficacy.

[0008] Therefore, the present invention provides a platform source technology based on a nano-drug delivery system that can increase therapeutic efficacy by improving the delivery ability of anticancer drugs to cancer tissues, which are target sites, and reduce side effects by minimizing the accumulation of anticancer drugs in normal tissues.

[0009] [Prior Art Literature]

[0010] [Patent Document]

[0011] Korean Patent No. 10-0497258

[0012] Korean Patent No. 10-1612255

[0013] Korean Patent No. 10-1544498

[0014] Korean Patent No. 10-1670249

[0015] Korean Patent No. 10-1768681

[0016] The present invention aims to provide an anticancer drug based on a drug delivery system having an ultrafine particle size that can treat at a lower dose compared to finished anticancer drugs used in existing clinical trials, has no effect on normal tissues, thus improving patient compliance and reducing side effects during or after treatment.

[0017] In addition, the present invention aims to provide an anticancer agent based on a drug delivery system having an ultrafine particle size that is highly useful in clinical practice, enabling effective progress of anticancer treatment by controlling the number of drug administrations according to the course of treatment by minimizing or overcoming the incidence of multidrug resistance due to repeated administration.

[0018] In addition, the present invention aims to provide an anticancer drug based on a drug delivery system having an ultrafine particle size that is excellent in convenience of administration, as anticancer treatment is effectively carried out not only through intravascular administration but also through oral administration, thereby saving time by not having to go to the hospital except on the date of the desired prescription, improving the quality of life of the patient, and even if side effects occur, they disappear quickly when the administration is stopped.

[0019] The present invention comprises the steps of: preparing a first mixture by mixing polysorbate, soybean oil, and an anticancer drug;

[0020] A step of mixing the first mixture and the first polymer and heating to 50 to 70°C to prepare a second mixture;

[0021] A step of cooling the second mixture to produce a first solid;

[0022] A step of preparing a third mixture by mixing the first solid with a first solvent; and

[0023] A step of manufacturing a nano-platform loaded with an anticancer drug by freeze-drying the third mixture,

[0024] A method for manufacturing a nanoplatform loaded with an anticancer drug is provided.

[0025]

[0026] In addition, the present invention is a nanocomposite having a micelle structure,

[0027] The inner core of the above micelle structure contains polysorbate and soybean oil in which an anticancer drug is dissolved,

[0028] The outer shell of the above micelle structure comprises a first polymer,

[0029] Provides a nano platform loaded with anticancer drugs.

[0030] In the present invention, a nano-platform loaded with an anticancer drug can be provided by a simple method based on the thermal phase transition phenomenon of a polymer without using any anhydrous ethanol or organic solvent.

[0031] The present invention provides a nano-platform for drug delivery that is loaded with a taxane-type anticancer drug and has an ultrafine particle size in an aqueous solution.

[0032] In addition, the present invention can provide a nano-platform for drug delivery with improved particle stability in an aqueous solution compared to a similar finished anticancer drug to which a drug delivery technology currently being used in clinical practice is applied, and the nano-platform can have an excellent effect of delivering an anticancer drug into cells even in cancer cells that have drug resistance to the anticancer drug.

[0033] Furthermore, nanoplatforms loaded with anticancer drugs administered intravenously can be naturally excreted through the bladder and urethra, preventing accumulation in unwanted organs or tissues, thereby reducing toxicity. Furthermore, nanoplatforms loaded with anticancer drugs can be absorbed into the body through oral as well as intravenous administration.

[0034] Figure 1 shows a schematic diagram of the manufacturing of a nano-platform loaded with an anticancer drug using a thermal phase transition phenomenon.

[0035] Figure 2 is a graph showing the particle size distribution in an aqueous solution of (a) a nanoplatform loaded with paclitaxel and (b) a nanoplatform loaded with docetaxel.

[0036] Figure 3 shows prototype photographs of (a) a nanoplatform loaded with paclitaxel and (b) a nanoplatform loaded with docetaxel.

[0037] Figure 4 is a graph comparing the release patterns of anticancer drugs released from a nanoplatform loaded with paclitaxel, Taxol Inj. (Samyang Biopharm), a liquid-type finished anticancer drug, and Genexol PM Inj. (Samyang Biopharm), a freeze-dried finished anticancer drug.

[0038] Figure 5 is a graph comparing the release patterns of anticancer drugs released from a nanoplatform loaded with docetaxel, a liquid-type finished anticancer drug, Taxotere Inj. (Sanofi Aventis), and a freeze-dried finished anticancer drug, Nanoxel M Inj. (Samyang Biopharm).

[0039] Figure 6 is a photograph showing the change in particle size over time at 25°C after dispersing each of the nanoplatform loaded with paclitaxel and Genexol PM Inj. in 0.9% NaCl, which is water for injection.

[0040] Figure 7 is a photograph showing the change in particles over time at 37°C after dispersing each of the nanoplatform loaded with paclitaxel and Genexol PM Inj. in 0.9% NaCl, which is water for injection.

[0041] Figure 8 is a comparative image using a fluorescence microscope showing the results of evaluating the intracellular uptake behavior of the nanoplatform loaded with docetaxel and a fluorescent material using (a) MCF-7 SEN and (b) MCF-7 ADR. In addition, it is a graph quantifying the amount of drug uptake over time using (c) MCF-7 SEN and (d) MCF-7 ADR.

[0042] Figure 9 is an image of a nanoplatform loaded with docetaxel and a fluorescent material administered to the tail vein of a normal nude mouse, and the behavior of the drug being excreted through the bladder over time was observed using a near-infrared fluorescence device.

[0043] Figure 10 is a graph quantifying the amount of fluorescence absorbed and present in the blood over time after oral administration of (a) a small-molecule fluorescent substance, indocyanine green (ICG), and (b) a nanoplatform loaded with docetaxel and a fluorescent substance to mice. Furthermore, Figure 10(c) is an image observing the degree of substance accumulation in major organs extracted 7 hours after substance administration.

[0044] Figure 11 is an image of a nanoplatform loaded with an anticancer drug and a fluorescent material applied to 3D cell cultured Caco-2 cells, and then observed using a fluorescence microscope in comparison with a tight junction-specific fluorescent material (FITC-phalloidin tight junction marker).

[0045] Figure 12 is a graph showing the pharmacokinetics over time after administering nanoplatforms loaded with paclitaxel, Taxol Inj and Genexol PM Inj., into the tail vein of mice.

[0046] Figure 13 is a graph showing the amount of paclitaxel absorbed in major organs extracted 24 hours after administering a nanoplatform loaded with paclitaxel intravenously and orally, respectively.

[0047] Figure 14 is a graph showing the results of evaluating single-dose toxicity after administering paclitaxel-loaded nanoplatforms, Taxol Inj and Genexol PM Inj., to the tail vein of mice at different drug doses.

[0048] Hereinafter, the present invention will be described in detail.

[0049] However, the present invention is susceptible to various modifications and may take various forms. The specific examples and descriptions described below are intended only to aid understanding of the present invention and are not intended to limit the present invention to any specific disclosed form. The scope of the present invention should be understood to include all modifications, equivalents, and alternatives falling within the spirit and technical scope of the present invention.

[0050] The terms "first" and "second" used herein do not denote an order, but rather are used to distinguish between the two components, and thus do not limit the two components. In particular, the terms "first mixture," "second mixture," and "third mixture" each refer to a mixture that includes the mixing elements presented before the appearance of each term, and the terms "first," "second," and "third" are used to distinguish the three mixtures.

[0051]

[0052] The present invention comprises a step of preparing a first mixture by mixing polysorbate, soybean oil, and an anticancer drug (hereinafter, the first mixture preparation step);

[0053] A step of mixing the first mixture and the first polymer and heating to 40 to 70°C to prepare a second mixture (hereinafter, the second mixture preparation step);

[0054] A step of cooling the second mixture to produce a first solid (hereinafter, the first solid production step);

[0055] A step of preparing a third mixture by mixing the first solid with a first solvent (hereinafter, the third mixture preparation step); and

[0056] The present invention relates to a method for manufacturing a nanoplatform loaded with an anticancer drug, comprising a step of manufacturing a nanoplatform loaded with an anticancer drug by freeze-drying the third mixture (hereinafter, the step of manufacturing a nanoplatform loaded with an anticancer drug).

[0057] The present invention improves the delivery capacity of anticancer drugs to target cancer tissues, thereby enhancing therapeutic efficacy and minimizing the accumulation of anticancer drugs in normal tissues, thereby reducing side effects. Specifically, the present invention utilizes paclitaxel or docetaxel, which are cytotoxic anticancer drugs of the taxane series, as an anticancer drug. The anticancer drug is loaded into a nanoplatform using a manufacturing method utilizing the polymer thermal phase transition phenomenon. When rehydrated, a nanoplatform is manufactured in which ultrafine, stably and uniformly dispersed particles are formed without precipitation of the water-insoluble anticancer drug. The anticancer drug-loaded nanoplatform according to the present invention enables treatment at lower dosages compared to finished anticancer drugs used in existing clinical trials. It does not affect normal tissues, thereby reducing side effects during or after treatment. In addition, it minimizes or overcomes the incidence of multidrug resistance due to repeated administration, allowing for control of the frequency of drug administration depending on the course of treatment. This enables high clinical utility and, furthermore, allows for the application of various drugs.

[0058] In the present invention, the "nanoplatform" is a nanotechnology-based drug delivery system that uses a formulation to deliver drugs to a desired body site or release them at an appropriate time. In other words, the goal is to maximize therapeutic efficacy and effectiveness by selectively delivering drugs to the target site and optimizing effective blood concentrations for a prolonged period of time, depending on the disease, while minimizing adverse drug reactions.

[0059] In the present invention, the "anticancer drug-loaded nanoplatform" is a micelle-structured nanocomposite capable of loading an anticancer drug into the inner core of the micelle structure. Here, the anticancer drug is a raw anticancer drug, and the nanoplatform loaded with the anticancer drug can be used as a finished product (complete anticancer drug).

[0060] In the present invention, the “first mixture preparation step” is a step of forming a first mixture by mixing polysorbate, soybean oil, and an anticancer drug.

[0061] In one specific example, polysorbate is a component that has both hydrophilic and hydrophobic properties and is utilized as a surfactant in pharmaceuticals or cosmetics. The polysorbate also has the characteristic of being a solubilizing agent that dissolves water-insoluble active ingredients.

[0062] In one specific embodiment, the polysorbate may be selected from polysorbate 20 or polysorbate 80.

[0063] In one specific example, soybean oil has the property of being completely water-insoluble and thus does not mix with water. During the emulsification process, it has the characteristic of being loaded into particles through hydrophobic bonding with insoluble active ingredients. In the present invention, soybean oil can be used to load anticancer drugs into nanoplatforms.

[0064] The content of the above soybean oil may be 0.5 to 15 parts by weight, 0.5 to 5 parts by weight, or 1 to 1.5 parts by weight relative to 100 parts by weight of polysorbate.

[0065] In one specific example, the anticancer drug may be a taxane-based anticancer drug, which is a water-insoluble or poorly soluble cytotoxic anticancer drug. Specifically, the taxane-based drug may include one or more selected from the group consisting of paclitaxel, docetaxel, sorafenib, and doxorubicin.

[0066] The term "anti-cancer" may mean inhibiting the progression of cancer or alleviating cancer in an animal, preferably a mammal, more preferably a human, having cancer.

[0067] The content of the above anticancer drug may be 1 to 5 parts by weight or 3 to 4 parts by weight relative to 100 parts by weight of polysorbate. Within this content range, the anticancer drug can be stably dispersed in the mixture and easily loaded into the nanoplatform. The above mixing ratio is an example of a mixing ratio that can achieve the aforementioned effects intended by the present invention.

[0068] In one specific embodiment, the first mixture may further comprise a fluorescent material in addition to the aforementioned components. In the present invention, the combination of an anticancer drug and a fluorescent material may enable imaging of the nanoplatform.

[0069] The above fluorescent material can be an insoluble fluorescent material. The above fluorescent material can penetrate / absorb into tissues and can be applied clinically.

[0070] The content of the above fluorescent material can be appropriately adjusted depending on the intended use.

[0071] In one specific example, the first mixture can be prepared as a solution by mixing the aforementioned components. The first mixture has the characteristics of being transparent and uniform without any foreign matter.

[0072] In the present invention, the “second mixture preparation step” is a step of preparing a second mixture by mixing the first mixture and the first polymer and then heating the mixture to 50 to 70°C.

[0073] In the present invention, the first polymer may be a poloxamer, and the poloxamer may include at least one selected from the group consisting of poloxamer 188 and poloxamer 407.

[0074] In one specific example, poloxamer is a triblock copolymer in which hydrophilic groups and hydrophobic groups are combined. The poloxamer may have a structure of Poly(oxyethylene)-poly(oxypropylene)-poly(oxyethylene) [POE-POP-POE]. Poloxamer is solid at room temperature, soluble in water and ethanol, and poloxamer solution is liquid at low temperatures, but its viscosity increases as the temperature increases, and when the concentration is high, it exhibits thixotropy of a sol-gel depending on the temperature. In addition, the poloxamer does not damage the mucosal cell membrane.

[0075] The above poloxamer can be used, such as Coliphor P188 and P407, which are commercially available. Coliphor P188 and P407 are applicable to clinical practice. For example, Coliphor P188 (POE) a -(POP) b -(POE) c It refers to a poloxamer having a molecular weight of approximately 8350, which is a compound in which b is 30 and the sum of a and c is approximately 75 in the structure.

[0076] In general, the introduction of a hydrophilic group is essential to provide functionality that allows the substance and active ingredient to be uniformly and stably dispersed without aggregation for a long period of time in an aqueous solution, and additionally, to provide important functionality that increases body circulation when administered into the blood vessels to deliver the active ingredient to the desired disease site.

[0077] In the present invention, the first polymer may additionally include at least one selected from the group consisting of vitamin E TPGS and solutol HS 15 together with poloxamer. The compound may function as a stabilizer.

[0078] Solutol is an amphoteric polymer with both hydrophilic and hydrophobic properties, used as a nonionic dissolving agent or emulsifier. It is composed of 15 moles of ethylene oxide (the main component of polyethylene oxide (PEG)) and 1 mole of 12-hydroxystearic acid, and its main chain contains a lipophilic stearic structure, allowing it to exhibit excellent mixing properties with hydrophobic drugs. The solutol can be used as Solutol HS-15.

[0079] Additionally, vitamin E TPGS is a vitamin E derivative in which a polyethylene glycol subunit is attached to the ring hydroxyl of the vitamin E molecule by a succinic acid diester. Vitamin E TPGS stands for D-α-tocopherol polyethylene glycol 1000 succinic acid (MW=530). TPGS is a nonionic surfactant.

[0080] In one specific example, the content of the first polymer may be 20 to 70 parts by weight or 50 to 70 parts by weight relative to the total weight of the first mixture.

[0081] In one specific example, the heating temperature may be 50 to 70°C or 50 to 60°C. If the heating temperature is lower than 50°C, there is a problem in that the first polymer does not melt because it is lower than the melting point of the first polymer, and thus cannot be mixed with the first mixture. If the heating temperature exceeds 70°C, there is a problem in that the first polymer and the anticancer drug decompose.

[0082] In one specific example, the second mixture is a solution in which the first polymer is added to a mixture of the aforementioned components, the first polymer, soybean oil, and an anticancer drug, and this may also have transparent and uniform characteristics.

[0083] In the present invention, the “first solid preparation step” is a step of preparing a first solid by cooling the second mixture.

[0084] In one specific example, the cooling temperature may be 15 to 30°C or 15 to 25°C. The second mixture is prepared by reacting the first polymer, specifically, the poloxamer, at a temperature higher than the melting point thereof, and then cooled to solidify the mixture. During this process, the first polymer encapsulates the anticancer drug, thereby loading the anticancer drug into the interior of the particle. If the cooling temperature exceeds 30°C, the second mixture is cooled slowly, which reduces the crystallization of the particles, and thus there is a problem in that the uniformity of the particle size is reduced. In the present invention, by setting the cooling temperature to 15 to 30°C, a nano-sized nanoplatform solid can be easily prepared without using a medium for rapid cooling.

[0085] In the present invention, the first solid can be manufactured through the above steps.

[0086] In the present invention, the “third mixture preparation step” is a step of preparing a third mixture by mixing a first solvent into the first solid material.

[0087] In one specific example, the first solvent may be tertiary distilled water. The first solid may be dissolved in the first solvent, and the active ingredient or other substances included therein may be uniformly and stably dispersed without causing agglomeration or foreign matter sensation.

[0088] The present invention is characterized by not using organic solvents. If organic solvents are used, there is a risk that the form of the formulation (nanoplatform) loaded with the anticancer drug will collapse, rendering it unusable. Furthermore, even if collapse does not occur, a step to remove the organic solvent is essential, and there is a risk of toxicity in the body due to residual solvents.

[0089] In one specific example, using tertiary distilled water as a solvent eliminates the possibility of solvent-induced toxicity during the manufacturing process, resulting in high biocompatibility. Furthermore, this method offers a distinct advantage over conventional organic solvent-based methods (self-assembly emulsification), which are widely used to manufacture liposomes or polymer-based nanoplatforms.

[0090] In one specific embodiment, a third mixture may be formed through the above steps. The third mixture may have uniform characteristics.

[0091] The present invention may further include a step of removing impurities from the third mixture after performing the third mixture preparation step.

[0092] In one specific embodiment, the method for removing impurities is not particularly limited, and any method applicable in the technical field to which the present invention pertains may be applied. For example, impurities may be removed using a 0.2 to 1.2 μm filter, a 0.5 to 1.1 μm filter, a 0.6 to 1.0 μm filter, a 0.7 to 0.9 μm filter, or a 0.8 μm filter.

[0093] In the present invention, the “step of manufacturing a nanoplatform loaded with an anticancer drug” is a step of manufacturing a nanoplatform loaded with an anticancer drug by freeze-drying the third mixture from which the impurities have been removed.

[0094] In one specific example, freeze-drying is a method of removing moisture by freezing an object and then lowering the surrounding air pressure to control the sublimation of solid water into gas. While the freeze-drying method is not specifically described, it is self-evident that any freeze-drying method applicable to the technical field to which the present invention pertains can be applied.

[0095] In one specific example, through the above steps, a nanoplatform loaded with a solid-form anticancer drug can be manufactured.

[0096] The above "nano" is expressed as "ultra-fine" and may refer to a size of 1 to 1,000 nm. The nanoplatform may be a delivery vehicle or vehicle capable of loading the aforementioned anticancer drug.

[0097]

[0098] In addition, the present invention relates to a nanoplatform loaded with an anticancer drug manufactured by the above-described manufacturing method.

[0099] The nanoplatform loaded with an anticancer drug according to the present invention is a nanocomposite with a micelle structure,

[0100] The inner core of the above micelle structure contains polysorbate and soybean oil in which an anticancer drug is dissolved,

[0101] The outer shell of the above micelle structure may include a first polymer.

[0102] In one specific example, the nanoplatform loaded with an anticancer drug may have an average diameter of 1 to 500 nm, 1 to 100 nm, 1 to 90 nm, 1 to 80 nm, 1 to 70 nm, 1 to 60 nm, 1 to 50 nm, 1 to 40 nm, 1 to 30 nm, 1 to 20 nm, 3 to 15 nm, 5 to 12 nm, 7 to 10 nm, or 8 to 9 nm. If the average diameter of the nanoplatform loaded with an anticancer drug is too large, it is unsuitable for use as a drug delivery vehicle. That is, as described above, the present invention provides a nanoplatform that can be applied for drug delivery by controlling the average diameter of the nanoplatform loaded with an anticancer drug to be ultrafine, and has improved particle stability in an aqueous solution, has drug resistance to an anticancer drug, and has an excellent delivery effect of an anticancer drug into cancer cells.

[0103] The nanoplatform loaded with the anticancer drug according to the present invention can be naturally excreted through the bladder and urethra, preventing accumulation in unwanted organs or tissues, thereby reducing toxicity. Furthermore, the nanoplatform loaded with the anticancer drug offers the advantage of being able to be administered orally as well as intravenously.

[0104] In one specific example, the cancer that is the target of treatment with the anticancer drug is not particularly limited and may include bladder cancer, breast cancer, stomach cancer, lung cancer, ovarian cancer, thyroid cancer, cervical cancer, central nervous system cancer, glioblastoma, liver cancer, skin cancer, pancreatic cancer, stomach cancer, colon cancer, rectal cancer, esophageal cancer, kidney cancer, lung cancer, epithelial cancer, blood cancer, prostate cancer, soft tissue sarcoma, multiple sclerosis, etc.

[0105]

[0106] In addition, the present invention relates to a pharmaceutical preparation comprising a nano-platform loaded with the aforementioned anticancer drug.

[0107] In one embodiment, the formulation may be administered intravenously or orally.

[0108] In one specific embodiment, the pharmaceutical formulation may include a pharmaceutically acceptable excipient, such as a pharmaceutically acceptable carrier for the nanoplatform. Generally, the therapeutically effective amount may vary depending on the anticancer drug included in the nanoplatform, the disease state being treated, the severity of the disease being treated, the age and relative health of the subject, the route and form of administration, and the judgment of the attending physician or veterinarian.

[0109] In the present invention, the "effective amount" of a nanoplatform or anticancer drug is an amount effective in bringing about a desired physiological change in a subject to which the nanoplatform is administered. A "therapeutically effective amount" refers to an amount of an anticancer drug or nanoplatform that elicits a biological or medical response, such as alleviation of symptoms of a disease or disorder being treated, or treatment or prevention of a disease or disorder, in a subject to which the nanoplatform of the present invention is administered.

[0110] In one embodiment, for oral administration, the nanoplatform of the present invention can be formulated into a wide variety of formulations. Pharmaceutically acceptable carriers can be solid or liquid. Solid-form formulations can include granules, tablets, pills, lozenges, capsules, caseins, and suppositories. Solid carriers can also include one or more substances that can also function as diluents, flavoring agents, lubricants, binders, preservatives, tablet disintegrants, or encapsulating materials, for example. In tablets, the nanoplatform of the present invention is generally mixed with a carrier having the necessary binding capacity in an appropriate ratio and compressed into a desired shape and size. Suitable carriers can include magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethyl cellulose, low-melting wax, cocoa butter, and the like.

[0111] The nanoplatform of the present invention can also be administered parenterally, for example, by injection or infusion, for example, by intravenous, intraarterial, intraosseous, intramuscular, intracerebral, intracerebroventricular, intrasynovial, intrasternal, intrathecal, intralesional, intracranial, intratumoral, intradermal and subcutaneous injection or infusion.

[0112] Accordingly, the pharmaceutical composition of the present invention may be a sterile injectable or infusible preparation, for example, in the form of a sterile aqueous or oily suspension of the nanoplatform. This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents (e.g., Tween 80) and suspending agents. The sterile injectable or infusible preparation may also be a sterile injectable or infusible suspension in a non-toxic, parenterally acceptable diluent. Other examples of acceptable vehicles that may be used in the composition of the present invention include, but are not limited to, mannitol, water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils may be used as a suspending medium.

[0113] Specifically, the nanoplatform loaded with the anticancer drug of the present invention can be applied to the treatment of tumors by uniformly dispersing it in a solution suitable for the purpose (injection water, buffer solution, distilled water, etc.) and then administering it intravenously.

[0114] As examples of the injection solution, a 0.5% to 1.3% NaCl solution, a 0.6% to 1.2% NaCl solution, a 0.7% to 1.1% NaCl solution, a 0.8% to 1.0% NaCl solution, or a 0.9% NaCl solution can be used.

[0115]

[0116] Hereinafter, the present invention will be described in detail through examples. The following examples are intended to illustrate the present invention and are not intended to limit its scope. These examples are provided to ensure a complete disclosure of the present invention and to fully inform those skilled in the art of the invention of its scope. The present invention is defined solely by the scope of the claims.

[0117]

[0118] Example

[0119] Example 1. Fabrication of a nanoplatform loaded with an anticancer drug using the thermal phase transition phenomenon of a polymer.

[0120] Soybean oil and 0.6 g of Tween 80 (wherein, soybean oil is included in an amount of 0.5 to 5 parts by weight based on 100 parts by weight of Tween 80) were added to the reactor, and the mixture was sufficiently stirred at room temperature to obtain a transparent and uniform state (preparation of a first mixture). Next, 0.4 g of Kolliphor P188 was added, and the mixture was heated to 50 to 55°C and mixed by sufficient stirring to obtain a transparent, uniform, and slightly viscous molten liquid (preparation of a second mixture). The second mixture was cooled to obtain a first solid.

[0121] The above first solid was completely dispersed in triple-distilled water (18.2 mΩ or more) (preparation of a third mixture), sterilized by 0.2 μm filtration, and then the filtered solution was freeze-dried to obtain a stable solid-form nanoplatform having an average particle diameter of 10 nm when hydrated.

[0122]

[0123] Figure 1 shows a schematic diagram of the manufacturing of a nanoplatform loaded with an anticancer drug.

[0124] In the present invention, depending on the type of anticancer drug loaded on the nano platform, for example, when paclitaxel is used as the anticancer drug, it can be expressed as a nano platform loaded with paclitaxel.

[0125]

[0126] Example 2. Preparation of a nanoplatform loaded with paclitaxel

[0127] A paclitaxel-loaded nanoplatform was manufactured by the method of Example 1, except that 20 mg of paclitaxel was additionally added during the preparation of the first mixture.

[0128]

[0129] Example 3. Preparation of a nanoplatform loaded with docetaxel

[0130] A nanoplatform loaded with docetaxel was manufactured by the method of Example 1, except that 20 mg of docetaxel was additionally added during the preparation of the first mixture.

[0131]

[0132] Experimental Example 1. Physical Characteristics of a Nanoplatform Loaded with Anticancer Drugs

[0133] The physical properties of the nanoplatform loaded with paclitaxel manufactured in Example 2 and the nanoplatform loaded with docetaxel manufactured in Example 3 were evaluated.

[0134]

[0135] Figure 2 is a graph showing the particle size distribution of (a) a nanoplatform loaded with paclitaxel and (b) a nanoplatform loaded with docetaxel in an aqueous solution. The particle size distribution was measured using a nano particle size analyzer.

[0136] As shown in the above Figure 2, it can be confirmed that the nanoplatform loaded with paclitaxel and docetaxel remains stably dispersed in an aqueous solution without aggregation of the drugs or particles over time.

[0137]

[0138] In addition, Fig. 3 shows prototype photographs of (a) a nano platform loaded with paclitaxel and (b) a nano platform loaded with docetaxel.

[0139] Hereinafter, the nano platform loaded with paclitaxel can be expressed as ON:Taxol Inj. or ON:Taxol Inj., and the nano platform loaded with docetaxel can be expressed as ON:D Taxol Inj. or ON:D Taxol Inj.

[0140]

[0141] Table 1 below shows the results of measuring the loading rates of paclitaxel and docetaxel in ON:Taxol Inj. and ON:D Taxol Inj. The drug loading was analyzed by HPLC (high-performance liquid chromatography).

[0142]

[0143] Classification Anticancer drug Drug loading rate (%) ON: Taxol Inj. Paclitaxel 96.22 ± 1.64 ON: D Taxol Inj. Docetaxel trihydrate 96.57 ± 0.75 Docetaxel anhydrous 97.46 ± 1.07

[0144] As shown in Table 1 above, it can be confirmed that the loading amount of paclitaxel and paclitaxel is 96% or more.

[0145]

[0146] Figure 4 shows a graph comparing the release patterns of anticancer drugs released from a nanoplatform loaded with paclitaxel (prototype name: ON:Taxol Inj.), a liquid-type finished anticancer drug Taxol Inj. (Samyang Biopharm), and a freeze-dried finished anticancer drug Genexol PM Inj. (Samyang Biopharm). Taxol Inj. is a liquid-type finished anticancer drug based on a dissolving agent (surfactant + absolute ethanol) used clinically, and Genexol PM Inj. is a freeze-dried finished anticancer drug using DDS technology.

[0147] As shown in the above Figure 4, it can be confirmed that the anticancer drug loaded on the nano platform according to the present invention can be continuously released in an aqueous solution.

[0148]

[0149] Figure 5 shows a graph comparing the release patterns of anticancer drugs released from a nanoplatform loaded with docetaxel (prototype name: ON:D Taxol Inj.), a liquid-type finished anticancer drug product, Taxotere Inj. (Sanofi Aventis), and a freeze-dried-type finished anticancer drug product, Nanoxel M Inj. (Samyang Biopharm). Taxotere Inj. is a liquid-type finished anticancer drug product based on a dissolving agent (surfactant + absolute ethanol) used clinically, and Nanoxel M Inj. is a freeze-dried-type finished anticancer drug product using DDS technology.

[0150] As shown in the above Figure 5, it can be confirmed that the anticancer drug loaded on the nanoplatform according to the present invention can be continuously released in an aqueous solution, and it can be confirmed that it has a similar shape to the nanoplatform loaded with paclitaxel.

[0151]

[0152] Experimental Example 2. Evaluation of particle stability in an injectable solution of a nanoplatform loaded with an anticancer drug.

[0153] The nanoplatform loaded with the anticancer drug according to the present invention must be uniformly and stably dispersed in the body and must maintain the particle size without significant change for a certain period of time in order to reach the desired site, that is, cancer tissue.

[0154] In order to administer the nanoplatform into the body, the solid form of the finished drug product, i.e. the nanoplatform, is dispersed in the injection solution, so the stability of the nanoplatform in the injection solution is very important.

[0155] Therefore, the paclitaxel-loaded nanoplatform (prototype name: ON:Taxol Inj) manufactured in Example 2 and Genexol PM Inj. (Genexol PM), a freeze-dried finished anticancer drug using DDS technology, were dispersed in 0.9% NaCl intravenous injection solution, and changes at room temperature (25°C) and body temperature (37°C) were confirmed.

[0156]

[0157] The results are shown in Figures 6 and 7.

[0158] Figure 6 is a photograph showing the change in particle size over time at 25°C after dispersing each of the nanoplatform loaded with paclitaxel and Genexol PM Inj. in 0.9% NaCl, which is water for injection.

[0159] In addition, Fig. 7 is a photograph showing the change in particles over time at 37°C after dispersing each of the nano platform loaded with paclitaxel and Genexol PM Inj. in 0.9% NaCl, which is water for injection.

[0160] As shown in Figures 6 and 7, the nanoplatform loaded with paclitaxel according to the present invention was confirmed to be stably dispersed without particle aggregation at both room temperature (25°C) and body temperature (37°C). However, Genexol PM Inj., a freeze-dried finished anticancer drug using DDS technology currently in clinical use, was confirmed to exhibit particle aggregation and cloudiness over time.

[0161]

[0162] Example 4. Preparation of a nanoplatform loaded with docetaxel and a fluorescent material.

[0163] A nanoplatform loaded with docetaxel was manufactured by the method of Example 1, except that 20 mg of docetaxel and 1 mg of fluorescent material were additionally added during the preparation of the first mixture.

[0164] The above fluorescent materials can be used for imaging in cell and animal models.

[0165]

[0166] Experimental Example 3. Evaluation of the intracellular uptake behavior of a nanoplatform loaded with an anticancer drug using cancer cells with multi-drug resistance (MDR) at the cellular level.

[0167] In general, finished anticancer drugs used in clinical practice work well in the early stages of treatment, but their therapeutic efficacy decreases due to drug resistance. Therefore, overcoming drug resistance is very important for effective treatment.

[0168] In this experimental example, the nanoplatform loaded with docetaxel and a fluorescent material manufactured in Example 4 was evaluated to determine whether it could overcome drug resistance at the cellular level. Specifically, the nanoplatform was treated with normal cancer cells (MCF-7 SEN) and drug-resistant cancer cells (MCF-7 ADR), and changes were observed over time.

[0169]

[0170] The results are shown in Fig. 8.

[0171] Figure 8 is a comparative image using a fluorescence microscope showing the results of evaluating the intracellular uptake behavior of a nanoplatform loaded with docetaxel and a fluorescent material using (a) MCF-7 SEN and (b) MCF-7 ADR. Additionally, a graph quantifying the drug uptake over time using (c) MCF-7 SEN and (d) MCF-7 ADR is shown. Taxotere Inj. was used as a comparison group.

[0172] As shown in the above Figure 8, when the nano-platform loaded with docetaxel and a fluorescent material according to the present invention is treated on cells, it can be confirmed that it is well delivered to the cytoplasm regardless of the presence or absence of the characteristics of the cells having drug resistance.

[0173]

[0174] Experimental Example 4. Evaluation of the In Vitro Release Behavior of a Nanoplatform Loaded with an Anticancer Drug Administered in the Body

[0175] To reduce the side effects of finished pharmaceutical products administered into the body, it is important that the pharmaceutical products do not accumulate in unwanted normal organs or tissues.

[0176] In this experimental example, the nanoplatform loaded with docetaxel and fluorescent material manufactured in Example 4 was intravenously administered to nude mice and observed for a certain period of time using real-time imaging equipment for small animals (IVIS Spectrum In Vivo Imaging System, Caliper Life Sciences).

[0177]

[0178] The results are shown in Fig. 9.

[0179] Figure 9 is an image of a nanoplatform loaded with docetaxel and a fluorescent material administered to the tail vein of a normal nude mouse, and the behavior of the drug being excreted through the bladder over time was observed using a near-infrared fluorescence device.

[0180] As shown in Fig. 9, it can be confirmed that the nanoplatform loaded with the administered anticancer drug and fluorescent material spontaneously excretes through the bladder into the urethra.

[0181]

[0182] Experimental Example 5. Evaluation of the Absorption Potential of a Nanoplatform Loaded with an Anticancer Drug Through Oral Administration

[0183] Existing taxane-type finished anticancer drugs have minimal or very low oral absorption, making the more efficient intravenous route of administration preferable. The nanoplatform loaded with the anticancer drug of the present invention aims to be applicable to patients through oral administration, taking into account the characteristics of the nanoplatform.

[0184] In this experimental example, the nanoplatform loaded with docetaxel and a fluorescent material prepared in Example 4 was orally administered to rat animals. After 2 and 7 hours, the animals were sacrificed and major organs were removed. The degree of absorption / accumulation of the nanoplatform was then confirmed using real-time fluorescence analysis equipment (IVIS Spectrum In Vivo Imaging System, Caliper Life Sciences).

[0185]

[0186] The results are shown in Figure 10.

[0187] Figure 10 is a graph quantifying the amount of fluorescence absorbed and present in the blood over time after oral administration of (a) a small-molecule fluorescent substance, indocyanine green (ICG), and (b) a nanoplatform loaded with docetaxel and a fluorescent substance to mice. Furthermore, Figure 10(c) is an image observing the degree of substance accumulation in major organs extracted 7 hours after substance administration.

[0188] As shown in the above Figure 10, compared to the control group (con.) that was administered nothing and the group administered only a fluorescent substance (free ICG), it can be confirmed that the group administered with the nanoplatform loaded with docetaxel and a fluorescent substance according to the present invention (nano anticancer formulation) accumulates in major organs at a higher content. This confirms that the nanoplatform according to the present invention is capable of absorption through oral administration.

[0189]

[0190] Experimental Example 6. Evaluation of the Oral Absorption Mechanism of a Nanoplatform Loaded with an Anticancer Drug

[0191] The oral absorption mechanism of a nanoplatform loaded with an anticancer drug (docetaxel) and a fluorescent material was evaluated at the 3D cell level.

[0192] After 3D culture of 5 x 10^4 / well of small intestinal mucosal cells (Caco-2 cells) in trans-wells (BD Biosciences) of 24-well plate size, the absorption mechanism was confirmed by treating the surface with a nano-platform loaded with anticancer drugs and fluorescent materials dispersed in PBS (phosphate buffered saline, pH 7.4).

[0193]

[0194] The results are shown in Figure 11.

[0195] Figure 11 is an image of a nanoplatform loaded with an anticancer drug and a fluorescent material applied to 3D cell cultured Caco-2 cells, and then observed using a fluorescence microscope in comparison with a tight junction-specific dye (FITC-phalloidin tight junction marker) (con.).

[0196] As shown in Figure 11, in the untreated control group (con.), only green actin-stained images were observed. In contrast, in the group treated with the nanoplatform loaded with anticancer drugs and fluorescent substances (Drug / dye-loaded Nanoplatform), strong red fluorescence in a mesh-like shape was observed.

[0197] When the image stained using the FITC-phalloidin marker, which can stain the action of tight junctions, and the image stained by the nano-platform loaded with the anticancer drug and fluorescent substance are overlapped, it can be confirmed that the nano-platform loaded with the anticancer drug and fluorescent substance according to the present invention is capable of oral absorption through tight junctions.

[0198]

[0199] Experimental Example 7. Pharmacokinetic Evaluation through Intravenous Administration of a Nanoplatform Loaded with an Anticancer Drug

[0200] The paclitaxel-loaded nanoplatform (prototype name: ON:Taxol Inj.) manufactured in Example 3 and Taxol Inj. and Genexol PM Inj. were each administered through the tail vein of a rat animal at 1 mg / kg based on anticancer drugs, and the amount of drug in the blood over time was analyzed by LC / MS / MS.

[0201]

[0202] The results are shown in Fig. 12 and Table 2.

[0203] Figure 12 is a graph showing the pharmacokinetics over time after administration of a nanoplatform loaded with paclitaxel, Taxol Inj., a liquid-type finished anticancer drug, and Genexol PM Inj., a freeze-dried finished anticancer drug, into the tail vein of a mouse.

[0204]

[0205] Dose (mg / kg)T 1 / 2 (hr)AUC last (hr*ng / mL)CI_obs (mL / min / kg)MRT inf _obs (hr)Vss_obs (L / kg)Taxol Inj.1 mg / kg0.9 ± 0.639.2 ± 4.6334.2 ± 55.70.9 ± 0.517.9 ± 6.4Genexol PM Inj.0.6 ± 0.414.0 ± 5.5957.4 ± 423.80.8 ± 0.635.6 ± 13.5ON:Taxol Inj.2.7 ± 0.3136.6 ± 19.5111.6 ± 14.43.0 ± 0.219.9 ± 3.3

[0206] As shown in the above Figure 12, Taxol Inj. showed an intermediate blood drug level among the groups through the AUClast result, and Genexol PM Inj. was lower than Taxol Inj. In this regard, it can be confirmed that ON:Taxol Inj. according to the present invention has a significantly higher AUClast result.

[0207] The reason Genexol PM Inj. has the lowest AUClast is believed to be due to the difference in stability of particles dispersed in aqueous solution at body temperature, and temporarily increasing the AUClast of Taxol Inj. causes many problems, including systemic toxicity and anticancer resistance.

[0208]

[0209] Experimental Example 8. Bioavailability Evaluation of a Nanoplatform Loaded with an Anticancer Drug Through Oral Administration

[0210] Through the aforementioned experimental examples, we confirmed that the nanoplatform loaded with anticancer drugs can be absorbed orally. To develop it into a practical pharmaceutical, high bioavailability through oral administration is essential, thereby increasing its potential for bioapplication.

[0211] In this experimental example, the paclitaxel-loaded nanoplatform (prototype name: ON:Taxol Inj.) manufactured in Example 3 was administered intravenously (1 mg PTX / kg) and orally (10 mg PTX / kg) to rat animals, and pharmacokinetic parameter values ​​were obtained. In addition, the animals were sacrificed 24 hours later, and the amount of drug absorbed into major organs was analyzed by LC / MS / MS.

[0212] As a control group, Taxol Inj. and Genexol PM Inj. were administered intravenously (1 mg PTX / kg), respectively, and pharmacokinetic parameter values ​​were obtained.

[0213] In addition, among the control groups, Genexol PM Inj. had a low AUClast result, so the pharmacokinetic parameter values ​​of Taxol Inj. were used, and the oral bioavailability of the nanoplatform loaded with paclitaxel was calculated using Equation 1 below.

[0214]

[0215] <Formula 1>

[0216] F(%) = (PO, AUC value / PO, administered dose) / (IV, AUC value / IV, administered dose)*100

[0217]

[0218] The results are shown in Fig. 13 and Table 3.

[0219]

[0220] Distinction AUC last(hr*ng / mL) Oral bioavailability of Taxol injection compared to Taxol injection F(%)IVPOMaleFemaleMaleFemaleMaleFemaleTaxol injection (Samyang Biopharm) 39.2 ± 4.6 36.3 ± 11.6--25.74 26.83Genexol PM injection (Samyang Biopharm) 14.0 ± 5.5 9.6 ± 4.7--On:Taxol injection (proprietary source technology) 136.6 ± 19.5 131.4 ± 12.9 100.9 ± 25.4 97.4 ± 17.5

[0221] As shown in Figure 13 and Table 3, the drug distribution in major organs through intravenous and oral administration was confirmed, and it was confirmed that the paclitaxel component in the small intestine, which is effective for oral absorption, was more than three times higher than in the intravenous administration group.

[0222] Furthermore, the nanoplatform loaded with paclitaxel demonstrated a high oral bioavailability of approximately 26% on average. This is a remarkably high oral absorption rate, compared to the reported oral absorption rate of less than 1-2% for conventional paclitaxel.

[0223]

[0224] Experimental Example 9. Single-Dose Toxicity Evaluation of a Nanoplatform Loaded with Anticancer Drugs

[0225] The single-dose toxicity of the paclitaxel-loaded nanoplatform was evaluated according to the administered dose. Taxol Inj. and Genexol PM Inj. were used as controls.

[0226]

[0227] The results are shown in Figure 14.

[0228] As shown in Fig. 14, Taxol Inj., a liquid-type finished anticancer drug, showed the highest body toxicity, with approximately 80% of experimental animals dying in the high-dose group of 30 mg PTX / kg, regardless of male or female.

[0229] Genexol PM Inj. has a 20% mortality rate even at 50 mg PTX / kg, and a 60% mortality rate at approximately 2.5 times the dose, confirming that it has lower toxicity.

[0230] In comparison, it was confirmed that the nanoplatform loaded with paclitaxel according to the present invention had the lowest toxicity, and no deaths were confirmed even in the group administered 120 mg PTX / kg, which was 2.5 times higher than that of Genexol PM Inj.

Claims

1. A step of preparing a first mixture by mixing polysorbate, soybean oil, and an anticancer drug; A step of mixing the first mixture and the first polymer and heating them to 50 to 70°C to prepare a second mixture; A step of cooling the second mixture to produce a first solid; A step of preparing a third mixture by mixing the first solid with the first solvent; and A step of producing a nano platform loaded with an anticancer drug by freeze-drying the third mixture, Method for manufacturing a nanoplatform loaded with an anticancer drug.

2. In paragraph 1, A method for producing a nanoplatform loaded with an anticancer drug, wherein soybean oil is included in an amount of 0.5 to 15 parts by weight relative to 100 parts by weight of polysorbate.

3. In paragraph 1, A method for producing a nanoplatform loaded with an anticancer drug, wherein the anticancer drug comprises at least one selected from the group consisting of paclitaxel, docetaxel, sorafenib, and doxorubicin.

4. In paragraph 1, A method for producing a nanoplatform loaded with an anticancer drug, wherein the anticancer drug is included in an amount of 1 to 5 parts by weight relative to 100 parts by weight of polysorbate and soybean oil.

5. In paragraph 1, A method for producing a nanoplatform loaded with an anticancer drug, wherein the first polymer comprises at least one selected from the group consisting of poloxamer 188 and poloxamer 407.

6. In paragraph 5, A method for producing a nanoplatform loaded with an anticancer drug, wherein the first polymer further comprises at least one selected from the group consisting of vitamin E TPGS and solutol HS 15.

7. In paragraph 1, A method for manufacturing a nanoplatform loaded with an anticancer drug, wherein the content of the first polymer is 20 to 70 parts by weight relative to the total weight of the first mixture.

8. In paragraph 1, A method for manufacturing a nanoplatform loaded with an anticancer drug, wherein the first solvent is distilled water.

9. In paragraph 1, A method for manufacturing a nanoplatform loaded with an anticancer drug, further comprising a step of removing impurities from a third mixture.

10. As a nanocomposite with a micelle structure, The inner core of the above micelle structure contains polysorbate and soybean oil in which an anticancer drug is dissolved, The outer shell of the above micelle structure comprises a first polymer, Nanoplatform loaded with anticancer drugs.

11. In Article 10, A nanoplatform loaded with an anticancer drug, the size of the nanoplatform loaded with an anticancer drug being 1 to 500 nm.

12. In paragraph 10, A nanoplatform loaded with an anticancer drug, which is administered intravenously or orally.

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