Compositions and methods for treating solid tumors

Abstract

The present invention provides methods for treating solid tumors, reducing local tumor recurrence and metastatic spread of tumors, by administering pharmaceutical compositions comprising a particulate biodegradable substrate coated with a polymer / lipid matrix that provides local sustained release of a taxane chemotherapeutic agent directly to the tumor or into the tumor resection cavity. The present invention further provides methods for treating chemotherapy-resistant tumors.

Description

[Technical Field]

[0001] This application claims priority under U.S. Provisional Application No. 63 / 128218 filed December 21, 2020, U.S. Provisional Application No. 63 / 231662 filed August 10, 2021, and U.S. Provisional Application No. 63 / 243147 filed September 12, 2021. The entire contents of those priority applications are incorporated herein by reference.

[0002] The present invention generally relates to sustained-release compositions of chemotherapeutic agents, and their uses for the local treatment of solid tumors, and for the prevention of cancer recurrence and metastasis after resection. [Background technology]

[0003] Systemic therapy often fails because it is difficult to maintain a therapeutic level for a sufficient amount of time for the drug to effectively kill malignant tumors in and around the tumor. While this problem can be addressed by increasing the dose, there is disagreement about the trade-offs between efficacy, dose-escalation toxicity, and associated costs.

[0004] Systemic chemotherapy has inherent drawbacks, and therefore, the development of local drug delivery platforms is becoming increasingly active as a solution to enhance efficacy and reduce side effects.

[0005] Local drug delivery offers several advantages over systemic drug administration (such as oral or intravenous administration), making these drugs promising cancer treatments. Drug-eluting depots, for example, deliver high concentrations of the drug locally at the disease site while simultaneously reducing systemic peaks through sustained drug release. Furthermore, sustained local drug delivery systems provide a steady supply of the drug, improving disease outcomes and patient adherence to medication. Moreover, local drug delivery can reduce or even prevent systemic side effects often associated with systemic drug administration. These advantages make depots particularly promising in cancer therapy aimed at preventing tumor recurrence and metastasis, especially at the surgical margins where residual cancer cells remain after surgical resection; in such cases, depots can effectively target residual cancer cells at the surgical incision site through sustained drug delivery, with minimal or no detectable systemic side effects. Active efforts are being made to explore various local delivery technologies, including polymer biodegradable sustained-release systems. These systems typically take the form of microparticles or nanoparticles, and implantable films or patches, which have drawbacks such as burst and decaying release profiles. Gliadel®, one clinically approved therapy, utilizes a polyanhydride carrier (polyfeprozan), which allows for sustained release of carmustine into the extracellular fluid of the brain, thus eliminating the need for the drug to cross the blood-brain barrier. Disadvantages of biodegradable polyester and polyanhydride-based depot technologies include the relatively short drug release time available in many systems, as well as potential toxicity due to dose dumping (burst effect) and non-steady-state drug release. Gliadel®, for example, releases the majority of its drug within 5–10 days and exhibits a burst release in the first 12 hours (Brudn et al., Biomaterials, 178(2018)373–382). The initial burst release results in excessive local or systemic drug concentrations, and this burst effect further limits the total amount of drug that can be loaded onto the depot. Another significant drawback is that the released drug has poor penetration into brain tissue.Drug penetration using Gliadel® extends the distance from the resected tumor by a maximum of only 5 mm, and only for a short period of 1-2 days post-surgery (Dan Bunis et al., Efficacy of nanoparticle-encapsulated BCNU delivery in apCPP:SA scaffold for treatment of Glioblastoma Multiforme, 2012). U.S. Patent No. 9,956,172 discloses a multilayer drug delivery implant or wafer positioned near a living tissue to deliver drugs to it, particularly to deliver chemotherapeutic agents to the brain after brain tumor resection. The implant disclosed in U.S. Patent No. 9,956,172 includes a drug-containing layer, which includes the drug, lipids and hydrophilic polymers or pore-forming agents, and a hydrophobic coating agent including hydrophobic substances.

[0006] Glioblastoma multiforme (GBM) is one of the most common types of brain tumors and is invasive; it accounts for 50–60% of all brain tumors in humans and has a low median survival rate. GBM is generally characterized by high mortality, invasiveness, hypergrowth, and a poor prognosis. The current standard of treatment for patients with brain tumors is surgical resection of the tumor followed by chemotherapy (typically oral temozolomide) and radiation therapy, the two therapies of which are administered about one month after surgery. This delay allows for the initiation of the wound healing process. However, the difficulty of surgical resection and the severe adverse effects of radiation and chemotherapy hinder these approaches. In addition, a drawback of the aforementioned delay is that cancer cells continue to proliferate during this period.

[0007] Docetaxel is a cell division inhibitory taxane considered one of the most effective drugs for brain tumors, typically administered systemically by intravenous infusion. However, its high molecular weight and lipophilicity limit its anti-brain tumor activity; this is mainly due to limited transport across the blood-brain barrier and poor permeability across the blood-brain tumor barrier. Docetaxel is known to cause severe adverse events, including infection, neutropenia, hypersensitivity, thrombocytopenia, and neuropathy.

[0008] International patent application WO2010 / 007623 by one of the inventors of the present invention and other inventors discloses a drug delivery composition comprising a biodegradable polymer and a lipid-based matrix, which controls the release of an active ingredient; the contents of this reference are incorporated herein by reference. These drug delivery compositions make it possible to encapsulate one or more diverse biologically active molecules and release them at a pre-programmed rate for a period ranging from several days to several months.

[0009] In general, there is a need to develop safe and robust local anticancer therapies using taxanes, particularly docetaxel with reduced systemic toxicity; such local anticancer therapies are those that can increase the payload concentration at the tumor site, enhance permeability to target tumor cells, promote the eradication of tumor cells, reduce the probability of the tumor acquiring resistance at that point, and overcome drug resistance mechanisms. [Overview of the Initiative]

[0010] The present invention provides sustained-release anticancer compositions and methods for utilizing such compositions for local treatment of cancer, prevention of cancer recurrence, and inhibition of tumor metastasis.

[0011] In a first aspect of the present invention, a method for treating a solid tumor is provided, comprising administering a pharmaceutical composition comprising a microparticle biodegradable substrate coated with a polymer / lipid matrix containing taxen to a subject having a solid tumor. Application of the pharmaceutical composition to the tumor site induces local controlled release of the taxen drug at and around the tumor site over a predetermined period, preferably up to 10 weeks, thereby improving the therapeutic effect of the drug. According to some embodiments, the pharmaceutical composition is administered to the tumor resection site after tumor resection to kill residual cancer cells in the tumor resection cavity or near the resected tissue, thereby preventing local recurrence of cancer. According to some embodiments, the solid tumor is at least one of brain tumors, colon cancer, prostate cancer, lung cancer, pancreatic cancer, breast cancer, esophageal cancer, gastric cancer, head and neck cancer, and soft tissue sarcoma. According to certain embodiments, the solid tumor is a brain tumor selected from glioblastoma or glioblastoma multiforme, high-grade endogenous brain tumors, and brain metastases of other tumors. According to a particular embodiment, the brain tumor is glioblastoma multiforme.

[0012] In a second aspect of the present invention, a local sustained-release composition is provided, the local sustained-release composition comprising a polymer / lipid matrix, wherein a particulate biodegradable substrate is coated or embedded therein with a matrix containing encapsulated taxene, the composition stabilizes the taxene and slows the conversion of the taxane to a 7-epimer impurity during storage and further during its sustained-release period.

[0013] This invention is partly based on experimental results showing the following: In a solid tumor syngeneic mouse model of docetaxel-resistant colon cancer, a single intraoperative application of a docetaxel-containing sustained-release composition according to several embodiments of this invention after partial tumor resection resulted in a tumor-free survival rate of 75% at the end of the study (39 days post-surgery). In comparison, only 25% of the treatment group, which received 5 cycles of systemic docetaxel treatment, remained tumor-free, and there were no survivors in the untreated group. Furthermore, 25% of the mice treated with the composition experienced tumor recurrence at the end of the study, compared to 75% in the extensive systemic treatment group and 100% in the untreated group. Furthermore, the docetaxel sustained-release composition treatment group showed a 30-day delay in tumor recurrence after tumor resection, compared to only 9 days in both the systemic treatment group and the untreated control group; these were determined by the mortality rate for the first tumor in each group.

[0014] Furthermore, docetaxel sustained-release compositions according to specific embodiments of the present invention induced potent inhibition of tumor growth and recurrence in a partially resected human glioblastoma subcutaneous mouse model. A single topical application of the composition induced 98% inhibition of tumor growth compared to an untreated control (p<0.001) (41 days post-surgery); and 66% inhibition of tumor growth compared to multiple injections in the systemic chemotherapy group (p=0.0165). The 41-day survival rate for the docetaxel sustained-release composition was significantly higher than that of systemically treated mice or untreated mice (survival rates were 60%, 20%, and 10%, respectively).

[0015] Furthermore, in a rat model, the survival rate at 23 days after the start of treatment was 40% in the docetaxel composition applied near unresectable glioblastoma brain tumors, compared to 0% in the standard systemic treatment group (temozolomide 33.5 mg / kg, 5 days of treatment), the placebo group (docetaxel-free composition), and the untreated control group.

[0016] According to some embodiments of the present invention, a method for treating a solid tumor comprises administering to a subject having a solid tumor a pharmaceutical composition comprising: (a) Particulate biodegradable substrate; (b) Biodegradable polymers; (c) at least one phospholipid having a hydrocarbon chain of at least 12 carbon atoms; and (d) Takisen. In some embodiments, the pharmaceutical composition further comprises a sterol. In various embodiments, the taxene is selected from the group consisting of docetaxel, paclitaxel, derivatives of paclitaxel, and cabazitaxel. In a particular embodiment, the taxene is docetaxel. In some embodiments, the solid tumor is at least one of brain tumors, prostate cancer, lung cancer, pancreatic cancer, breast cancer, esophageal cancer, gastric cancer, head and neck cancer, and soft tissue sarcoma. In a particular embodiment, the solid tumor is a brain tumor selected from glioblastoma or glioblastoma multiforme and high-grade endogenous brain tumors. In a particular embodiment, the brain tumor is glioblastoma multiforme. In some embodiments, the tumor is a chemotherapy-resistant tumor. In some embodiments, the tumor is a taxane-resistant tumor.

[0017] According to some embodiments of the present invention, the present invention provides a method for reducing tumor cell regrowth at a solid tumor excision site, wherein the method comprises administering a pharmaceutical composition comprising the following to the solid tumor excision site: (a) Particulate biodegradable substrate; (b) Biodegradable polymers; (c) at least one phospholipid having a hydrocarbon chain of at least 12 carbon atoms; and (d) Takisen. In some embodiments, the pharmaceutical composition further comprises a sterol. In various embodiments, the taxene is selected from the group consisting of docetaxel, paclitaxel, derivatives of paclitaxel, and cabazitaxel. In a particular embodiment, the taxene is docetaxel. In some embodiments, the solid tumor is at least one of brain tumors, prostate cancer, lung cancer, pancreatic cancer, breast cancer, esophageal cancer, gastric cancer, head and neck cancer, and soft tissue sarcoma. In a particular embodiment, the solid tumor is a brain tumor selected from glioblastoma or glioblastoma multiforme and high-grade endogenous brain tumors. In a particular embodiment, the brain tumor is glioblastoma multiforme. In some embodiments, the tumor is a chemotherapy-resistant tumor. In some embodiments, the tumor is a taxane-resistant tumor.

[0018] According to several embodiments, the present invention provides a method for inhibiting tumor metastasis, wherein the method is applied to subjects having malignant solid tumors: (a) Particulate biodegradable substrate; (b) Biodegradable polymers; (c) at least one phospholipid having a hydrocarbon chain of at least 12 carbon atoms; and (d) Takisen, The method includes administering a pharmaceutical composition containing [a specific substance], thereby inhibiting tumor metastasis. In some embodiments, the pharmaceutical composition further comprises sterols. In various embodiments, the taxen is selected from the group consisting of docetaxel, paclitaxel, derivatives of paclitaxel, and cabazitaxel. In certain embodiments, the taxen is docetaxel. In some embodiments, the pharmaceutical composition is administered to the site of malignant tumor resection immediately after at least a portion of the malignant tumor has been surgically removed. In some embodiments, the solid tumor is at least one of brain tumors, colon cancer, prostate cancer, lung cancer, pancreatic cancer, breast cancer, esophageal cancer, gastric cancer, head and neck cancer, and soft tissue sarcoma. In certain embodiments, the solid tumor is a brain tumor selected from glioblastoma or glioblastoma multiforme, high-grade endogenous brain tumors, and intracranial metastases originating from other tumors. In certain embodiments, the brain tumor is glioblastoma multiforme. In some embodiments, the tumor is a taxane-resistant tumor.

[0019] Methods for treating solid tumors according to several embodiments of the present invention provide adjuvant cancer therapy. The pharmaceutical compositions described herein are intended to be administered locally to the tumor resection cavity immediately after tumor resection surgery for the purpose of increasing the survival rate of cancer patients. The pharmaceutical compositions of the present invention provide sustained, controlled local exposure to a taxane agent during tumor resection, enabling absorption and distribution of the taxane agent in the local environment of the resected tumor site, supplying taxane at therapeutic levels over a long period, thereby killing any remaining tumor cells at or near the tumor resection site, and reducing local tumor recurrence and metastatic spread of the tumor. The taxane is released from the pharmaceutical composition, but this release begins immediately after application to the tumor resection site and follows zero-order or near-zero-order kinetics. The taxane is released continuously for 2 to 10 weeks without an initial burst (less than 10% of the taxane encapsulated in the composition is released within the first 24 hours; typically, less than 8%, 7%, 6%, or 5% (w / w) of the taxane is released within the first 24 hours), thereby avoiding the potential for dose dumping (burst effect) toxicity.

[0020] The taxen drug is released locally for a period ranging from 2 to 10 weeks; 2 to 8 weeks; or 2 to 6 weeks; or 2 to 5 weeks; or 2 to 4 weeks, which is typically the time lag between surgical removal of the tumor and the initiation of adjuvant radiotherapy, chemotherapy, and / or biological treatment; all of these treatments are typically initiated only after the healing process of the surgical wound has begun. The disadvantage of delaying adjuvant treatment after tumor removal surgery is that cancer cells continue to proliferate and spread during this period. The methods and pharmaceutical compositions of the present invention overcome this disadvantage.

[0021] According to several embodiments, the present invention provides a neoadjuvant method for treating solid tumors, the method being: (a) Particulate biodegradable substrate; (b) Biodegradable polymers; (c) at least one phospholipid having a hydrocarbon chain of at least 12 carbon atoms; and (d) Takisen, The method includes injecting a pharmaceutical composition containing the above into a tumor. In some embodiments, the pharmaceutical composition further comprises sterols. In various embodiments, the taxen is selected from the group consisting of docetaxel, paclitaxel, paclitaxel derivatives, and cabazitaxel. In certain embodiments, the taxen is docetaxel. In some embodiments, the solid tumor is at least one of brain tumors, prostate cancer, lung cancer, pancreatic cancer, breast cancer, esophageal cancer, gastric cancer, head and neck cancer, and soft tissue sarcoma. The objective of neoadjuvant therapy is to reduce tumor size before surgical resection or radiotherapy, thereby simplifying the surgical procedure and reducing the risk of cancer cell spread during the surgical procedure. In some embodiments, the pharmaceutical composition may be injected directly into the tumor in dry powder form using an instrument suitable for dry powder injection. Alternatively, the pharmaceutical composition may be injected as a liquid suspension. In some embodiments, the tumor is a chemotherapy-resistant tumor. In some embodiments, the tumor is a taxane-resistant tumor.

[0022] In some embodiments, the particulate biodegradable substrate used in the pharmaceutical compositions and methods of the present invention consists of particles, which are typically spherical or spherical in shape. In some embodiments, the particles do not necessarily have to be spherical and / or steroidal, but are preferably spherical and / or spherical in shape, and their average diameter (as measured by laser diffraction) may be at least about 30 μm, at least about 40 μm, at least about 50 μm, at least about 60 μm, at least about 70 μm, at least about 80 μm, at least about 90 μm, at least about 100 μm, 30 μm to 120 μm, 30 μm to 100 μm, 50 μm to 100 μm, about 200 μm or less, about 180 μm or less, about 150 μm or less, about 140 μm or less, about 130 μm or less, about 120 μm or less, about 110 μm or less, or about 100 μm or less. Each possibility represents a separate embodiment of the present invention. In some embodiments, the particulate substrates used in the compositions and methods described herein are biocompatible, bioabsorbable, hydrophilic materials with low solubility in water, requiring a period of 4 weeks or more, 5 weeks or more, 6 weeks or more, 7 weeks or more, 8 weeks or more, 9 weeks or more, preferably 10 weeks or more, and further having a solid form at ambient temperature and being moldable. Any material having these properties can be used without limitation. In certain embodiments, the particulate substrate is composed of tricalcium phosphate (TCP), preferably β-TCP. In other embodiments, the particulate substrate is composed of polyvinyl alcohol (PVA), preferably PVA having at least 88% hydrolysis. In some embodiments, the particulate biodegradable substrate is neither calcium sulfate nor related hydrates such as calcium dihydrate or calcium sulfate hemihydrate. Although not limited by theory or mechanism of action, it is suggested that the polymer / lipid matrix coating the surface of biodegradable substrate particles protects the substrate particles from degradation by dissolution. The gradual elution of the substrate particles begins only when their surface is exposed to bodily fluids after degradation of the polymer / lipid matrix.The particle size is sufficiently large so that it does not move from the administration site until at least the majority, preferably all, of the drug has been released. The size of the biodegradable substrate must be large enough so that the pharmaceutical composition disclosed herein does not move from the application site. This is especially important when toxic drugs such as chemotherapeutic agents are released. Therefore, it is important that the overall shape of the particles does not change significantly during the drug release period. According to some embodiments, the pharmaceutical composition used decreases in total weight by about 10-15% during the taxane release period. The taxane-containing sustained-release composition is designed to be fixed to the tissue to prevent accidental migration to other compartments and organs over time. According to some embodiments, the particulate biodegradable substrate accounts for about 80-93% (w / w) of the total weight of the pharmaceutical composition.

[0023] The biodegradable polymer in the pharmaceutical composition according to embodiments of the present invention is a polyester. According to some embodiments, the polyester is selected from the group consisting of polylactic acid (PLA), polyglycolic acid (PGA), polylactic acid / glycolic acid copolymer (PLGA), and polycaprolactone and any combination or copolymer thereof. According to certain embodiments, the polyester is PLGA. According to some embodiments, the polyester component accounts for 0.5 to 5% (w / w) of the total weight of the pharmaceutical composition.

[0024] In some embodiments, the phospholipid comprises a fatty acid chain of at least 12 carbon atoms. In some embodiments, the fatty acid chain of the phospholipid comprises 18 or fewer carbon atoms. In some embodiments, the fatty acid chain of the phospholipid is fully saturated. In some embodiments, at least one of the phospholipid fatty acid chains is unsaturated (e.g., containing at least one double bond). In some embodiments, both of the phospholipid fatty acid chains are unsaturated. In some embodiments, a phospholipid having a hydrocarbon chain of at least 12 carbon atoms has a phase transition temperature of less than 60°C, less than 55°C, less than 50°C, less than 45°C, less than 42°C, less than 40°C, less than 38°C, less than 35°C, less than 32°C, less than 30°C, less than 28°C, or less than 25°C. In some embodiments, the phospholipid comprises a phospholipid selected from the group consisting of phosphatidylcholine, phosphatidylcholine mixtures, phosphatidylethanolamine, and combinations thereof. In some embodiments, the second lipid comprises phosphatidylcholine or a mixture of phosphatidylcholine. In some embodiments, the phosphatidylcholine is selected from the group consisting of DMPC, DPPC, DSPC, DOPC and any combination thereof. In some embodiments, the phosphatidylcholine is selected from the group consisting of DMPC, DPPC, DSPC and any combination thereof. In some embodiments, the phosphatidylcholine is selected from the group consisting of DMPC, DPPC and any combination thereof. In some embodiments, the phosphatidylcholine is selected from the group consisting of DMPC, DSPC and any combination thereof. In certain embodiments, the phosphatidylcholine is DMPC. In some embodiments, the phospholipid component accounts for 2 to 15% (w / w) of the total weight of the pharmaceutical composition.

[0025] In some embodiments, the pharmaceutical composition further comprises sterols. In some embodiments, the sterols are plant sterols. In some embodiments, the sterols are animal sterols. In certain embodiments, the sterols are cholesterol. In some embodiments, the sterols constitute 0-4% (w / w) of the total weight of the pharmaceutical composition. In some preferred embodiments, the sterols are cholesterol and constitute up to 50% (w / w) of the total lipid content of the pharmaceutical composition. Total lipid content refers to the total weight of all lipids contained in the pharmaceutical composition (e.g., sterols, phospholipids, and additional lipid additives contained in the pharmaceutical composition). In some embodiments, the sterols and polymers are linked non-covalently.

[0026] In some embodiments, the taxane is incorporated into a polymer / lipid-based matrix. In some embodiments, the taxane accounts for 0.2% to 2.6% (w / w) of the total weight of the pharmaceutical composition used in the method described herein. Alternatively, the taxane accounts for 0.5% to 1.5% (w / w) of the total weight of the pharmaceutical composition. In certain embodiments, the taxane accounts for 0.7% to 1.3% (w / w) or 0.7% to 1.0% (w / w) of the total weight of the pharmaceutical composition. In various embodiments, the taxane is selected from the group consisting of docetaxel, paclitaxel, paclitaxel derivatives, and cabazitaxel. In certain embodiments, the taxane is docetaxel. In some embodiments of the method of the present invention, the pharmaceutical composition is administered to the surface of a solid tumor or to the surface of a solid tumor resection cavity after surgical removal of the tumor. According to some embodiments of the method of the present invention, the pharmaceutical composition is applied to the surface of a solid tumor or the inner surface of an excision cavity, with a surface area of ​​1 cm². 2 The dose is administered in an amount ranging from 20 mg to 260 mg per unit area. According to another embodiment, the composition is 50 mg / cm³. 2 ~160 mg / cm³ 2 50 mg / cm³ 2 ~160 mg / cm³ 2 50 mg / cm³2 ~150 mg / cm 2 ; 50 mg / cm 2 ~120 mg / cm 2 ; 50 mg / cm 2 ~100 mg / cm 2 ; 50 mg / cm 2 ~100 mg / cm 2 ; 75 mg / cm 2 ~160 mg / cm 2 ; 75 mg / cm 2 ~120 mg / cm 2 ; 75 mg / cm 2 ~100 mg / cm 2 is applied in an amount within the range of.

[0027] According to some embodiments, the pharmaceutical composition is in powder form. According to some embodiments, the powder is spread on the tumor surface, or sprayed onto the tumor surface, or applied to the inner surface of the resection cavity. The powder may additionally or alternatively be intratumorally injected using a suitable powder injector. According to certain embodiments of the present invention, the pharmaceutical composition is formulated as a paste before being applied to the tumor site or the inner surface of the tumor in the resection cavity. According to some embodiments, the paste is spread on the tumor surface or applied to the inner surface of the resection cavity, for example, with a spatula. According to another embodiment, the pharmaceutical composition may be formulated as an injectable suspension.

[0028] Some embodiments of the solid tumor treatment method according to the present invention involve administering to a subject having a solid tumor a pharmaceutical composition comprising: (a) tricalcium phosphate particles; (b) a polyester; (c) phosphatidylcholine having a hydrocarbon chain of at least 12 carbons; and (d) a taxane, where the composition is intended to be locally administered to the solid tumor surface or the inner surface of the solid tumor resection cavity. In some embodiments, the composition further comprises cholesterol. In some embodiments, the taxane is selected from the group consisting of docetaxel, paclitaxel, derivatives of paclitaxel, and cabazitaxel. In certain embodiments, the taxane is docetaxel. In some embodiments, the polyester is PLGA (poly(lactic acid / glycolic acid copolymer)). In some embodiments, the hydrocarbon chain of phosphatidylcholine is saturated. In some embodiments, the phosphatidylcholine is 1,2-dimiristoyl-sn-glycero-3-phosphocholine (DMPC). In some embodiments, the docetaxel accounts for 0.2% to 2.6% (w / w) of the total weight of the pharmaceutical composition. Alternatively, the docetaxel accounts for 0.2% to 2.6% (w / w) of the total weight of the pharmaceutical composition. It accounts for 0.5% to 1.5% (w / w). According to a particular embodiment, the docetaxel accounts for 0.7% to 1.3% (w / w) or 0.7% to 1.0% (w / w) of the total weight of the pharmaceutical composition. According to some embodiments, the tricalcium phosphate (TCP) is selected from the group consisting of α-tricalcium phosphate, β-tricalcium phosphate and combinations thereof. According to a particular embodiment, the TCP is β-tricalcium phosphate. According to some embodiments, the pharmaceutical composition is distributed over a surface area of ​​1 cm². 2 Apply to the surface of a solid tumor or resection cavity in an amount ranging from 20 mg to 500 mg per unit area. According to another embodiment, 1 cm 2 The composition is applied in amounts ranging from 50mg to 400mg, 50mg to 350mg, 50mg to 300mg, 50mg to 275mg, 50mg to 250mg, 50mg to 225mg, 50mg to 200mg, 50mg to 180mg, 50mg to 170mg; 50mg to 160mg; 50mg to 150mg; 50mg to 120mg; 50mg to 100mg; 50mg to 100mg; 75mg to 160mg; 75mg to 120mg; and 75mg to 100mg per unit. According to some embodiments, the solid tumor is a brain tumor. According to some embodiments, the brain tumor is glioblastoma multiforme. According to some embodiments, the tumor is a taxane-resistant tumor.

[0029] In a particular embodiment, the present invention provides a method for treating a solid tumor, the method comprising topically administering a pharmaceutical composition comprising the following to the surface of the solid tumor or the surface of the solid tumor resection cavity: (a) 80-93% (w / w) tricalcium phosphate particles; (b) 1% to 4.0% (w / w) polyester; (c) 0.0-2.0% (w / w) cholesterol; (d) 4.0–15.0% (w / w) of phosphatidylcholine having at least 12 carbon hydrocarbon chains; (e) 0.2-2.6% (w / w) docetaxel. In some embodiments, the docetaxel accounts for 0.5% to 1.5% (w / w) of the total weight of the pharmaceutical composition. In certain embodiments, the docetaxel accounts for 0.7% to 1.3% (w / w) or 0.7% to 1.0% (w / w) of the total weight of the pharmaceutical composition. In some embodiments, the polyester is PLGA (poly(lactic acid / glycolic acid copolymer)). In some embodiments, the phosphatidylcholine hydrocarbon chain is saturated. In some embodiments, the phosphatidylcholine is 1,2-dimiristoyl-sn-glycero-3-phosphocholine (DMPC). In some embodiments, the tricalcium phosphate (TCP) is selected from the group consisting of α-tricalcium phosphate, β-tricalcium phosphate and combinations thereof. In certain embodiments, the TCP is β-tricalcium phosphate. In some embodiments, the pharmaceutical composition is distributed over a surface area of ​​1 cm². 2 Apply to the surface of a solid tumor or resection cavity in an amount ranging from 20 mg to 500 mg per unit area. According to another embodiment, 1 cm 2The composition is applied in amounts ranging from 50mg to 400mg, 50mg to 350mg, 50mg to 300mg, 50mg to 275mg, 50mg to 250mg, 50mg to 225mg, 50mg to 200mg, 50mg to 180mg, 50mg to 170mg; 50mg to 160mg; 50mg to 150mg; 50mg to 120mg; 50mg to 100mg; 50mg to 100mg; 75mg to 160mg; 75mg to 120mg; and 75mg to 100mg per unit. According to some embodiments, the solid tumor is a brain tumor. According to some embodiments, the brain tumor is glioblastoma multiforme. According to some embodiments, the tumor is a docetaxel-resistant tumor.

[0030] The pH of the pharmaceutical compositions disclosed herein is essentially determined by the additives in the pharmaceutical compositions. According to some embodiments, the pH of the pharmaceutical composition is 7.0 to 9.0, preferably 7.5 to 8.5, as measured with a pH electrode InLab® Solids Go-ISM. According to some embodiments, the pharmaceutical composition further comprises a pH adjuster. pH adjusters such as buffers or acids can be added to the pharmaceutical composition to maintain its pH at 3.5 to 7; 3.5 to 6.5; 4 to 6; 4 to 5.5; 4 to 5 or 4 to 4.5. Each possibility represents a distinct embodiment of the present invention. According to some embodiments, the taxene is stabilized by maintaining the pH of the pharmaceutical composition below 7, preferably below 6, more preferably between 4 and 5, thereby slowing the conversion of the taxene to its 7-epimer impurity during storage. According to certain embodiments, the taxene is docetaxel, and the pH of the pharmaceutical composition is between 4 and 5.5. Suitable acids to be included in the pharmaceutical composition include organic acids (such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and mixtures thereof) and inorganic acids (such as hydrochloric acid, phosphoric acid, nitric acid, and sulfuric acid, or combinations thereof). Acetic acid is a preferred pH adjuster. The amount of pH adjuster in a pharmaceutical composition according to several embodiments is 0.1-5% (w / w); 0.1-4% (w / w); 0.1-3% (w / w); 0.1-2% (w / w); 0.2-2% (w / w); 0.3-2% (w / w); 0.5-2% (w / w); 0.5-1.8% (w / w); 0.5-1.7% (w / w); 0.5-1.6% (w / w); 0.5-1.5% (w / w); 0.5-1.4% (w / w); 0.5-1.3% (w / w); 0.5-1.2% (w / w); 0.5-1.1% (w / w); or 0.5-1.0% (w / w) of the total weight of the pharmaceutical composition. Each possibility represents a distinct embodiment of the present invention.

[0031] A major challenge in chemotherapy is to deliver chemotherapy agents from the surface of a resected tumor into deep cancerous tissue. While it is possible to improve the efficacy of chemotherapy through active or passive targeted therapy with targeted drugs, or by enhancing penetration and retention (EPR), there is also the problem of the penetration of nanopharmaceuticals into the tumor stroma (Xiaoqian et al., Biomacromolecules 2019, 20:2637-48). To date, efficient penetration of active substances into tumor tissue has been unsuccessful in most treatment methods. This problem is even more serious in the treatment of brain tumors. Glioblastoma multiforme is a diffuse brain tumor characterized by high-level invasion into the brain parenchyma. This process is enhanced by interactions between local cells (small glial cells) and infiltrating immune cells (macrophages and Treg cells (regulatory T cells)), which produce cytokines and matrix-degrading enzymes that are important for tumor growth and expansion in the brain. Therefore, it is difficult, if not impossible, to completely remove (resect) GBM tumors by neurosurgery without imposing a significant risk of nerve damage on the patient. Consequently, despite continuous advances in neurosurgical techniques, the invasive behavior of GBM hinders complete tumor resection and is undoubtedly a major cause of poor clinical outcomes in patients. This invention provides three key factors that improve the penetration of the drug from the resection surface into the tissue: namely, (1) a high local concentration immediately adjacent to the surface of the tumor resection cavity; (2) prolonged exposure to the high concentration; and (3) physical protection of the released chemotherapeutic agent. Prolonged local high concentrations allow for higher concentrations of the released drug's activity, thereby not only extending drug exposure but also helping it penetrate deeper into the tissue, enabling the eradication of tumor cells that have infiltrated from the surface to deeper layers. According to several embodiments, the methods and compositions disclosed herein allow taxane penetration to extend to a distance of at least 0.5 cm from the surface of the resected tumor (e.g., the outer boundary of the residual tumor margin) as measured by quantitative autoradiography.In some embodiments, drug penetration extends to a distance of at least 0.6 cm, 0.7 cm, 0.8 cm, 0.9 cm, 1.0 cm, 1.2 cm, 1.3 cm, 1.4 cm, 1.5 cm, 1.6 cm, 1.7 cm, 1.8 cm, 1.9 cm, 2.0 cm, 2.1 cm, 2.2 cm, 2.3 cm, 2.4 cm, 2.5 cm, 2.6 cm, 2.7 cm, 2.8 cm, 2.9 cm, and 3.0 cm from the surface of the resected tumor. In some embodiments, drug penetration extends to a distance of 2.5 cm or more from the surface of the resected tumor, or to a distance of 2.4 cm, 2.3 cm, 2.2 cm, 2.1 cm, 2.0 cm, 1.9 cm, 1.8 cm, 1.7 cm, 1.6 cm, and 1.5 cm or more.

[0032] Taxanes are relatively large and highly hydrophobic, and this property limits their tissue penetration, with very few drugs reaching depths greater than 100 μm in tissues (Alastair H., Clin Cancer Res, 2007;13(9):2804-10). This is at least in part because free taxanes become highly (>98%) bound to circulating proteins, which limits their ability to penetrate tissues. The pharmaceutical compositions disclosed herein not only protect taxanes within the matrix during storage but also protect them upon release. When placed in an aqueous environment, the polymer / lipid matrix is ​​gently degraded, at which point the taxanes are released from the disclosed pharmaceutical compositions. It has been shown that the release of at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, and at least 90% of taxanes from the compositions disclosed herein is related to the lipid colloidal structure formed at the periphery of the outer layer of the lipid / polymer-based matrix upon exposure to an aqueous environment (e.g., body fluids). These lipid colloidal particles protect the drug from binding to circulating proteins but do not adversely affect drug uptake by tumor cells. While not limited by theory or mechanism of action, it is suggested that these lipid colloidal particles improve the penetration and infiltration of taxanes into tissues.

[0033] Further embodiments and the full scope of the applicability of the present invention will become apparent from the detailed description provided herein. However, since various modifications and changes within the spirit and scope of the present invention will be apparent to those skilled in the art from this detailed description, it should be understood that the detailed description and specific examples, while illustrating preferred embodiments of the present invention, are provided only as specific examples. [Brief explanation of the drawing]

[0034] [Figure 1] The following shows the docetaxel accumulation and release profiles from pharmaceutical compositions containing different phospholipids, some containing cholesterol and some not, according to multiple embodiments of the present invention. [Figure 2] The amount of docetaxel 7-epimer in a docetaxel sustained-release composition containing different cholesterol-containing or cholesterol-free phospholipids according to multiple embodiments of the present invention is shown. [Figure 3] The amounts of docetaxel 7-epimer in docetaxel sustained-release compositions containing different amounts of DMPC according to multiple embodiments of the present invention are shown. [Figure 4A] The effect of adding Tween-80 to a docetaxel sustained-release composition containing DMPC(4A) and DPPC(4B), according to a specific embodiment of the present invention, on the accumulation-release profile of docetaxel is demonstrated. [Figure 4B] The effect of adding Tween-80 to a docetaxel sustained-release composition containing DMPC(4A) and DPPC(4B), according to a specific embodiment of the present invention, on the accumulation-release profile of docetaxel is demonstrated. [Figure 5] The amounts of docetaxel 7-epimer in docetaxel sustained-release compositions containing various amounts of cholesterol, according to a specific embodiment of the present invention, are shown. [Figure 6] The profiles of paclitaxel accumulation and release from paclitaxel sustained-release compositions containing different phospholipids, according to a specific embodiment of the present invention, are shown. [Figure 7] This shows the accumulation and release of docetaxel from a docetaxel sustained-release composition containing either PLGA or PEG as a polymer component. [Figure 8] The mean tumor volume of CT26 colon cancer in BALB / c mice topically treated with various docetaxel sustained-release compositions according to specific embodiments of the present invention is shown. [Figure 9] The mean tumor volume of CT26 colon cancer in BALB / c mice treated topically with a docetaxel sustained-release composition according to a specific embodiment of the present invention, compared to systemic docetaxel treatment. [Figure 10] This study demonstrates the dose-response to topical treatment with a docetaxel sustained-release composition containing 0.87% (w / w) docetaxel, reflected in the mean tumor volume of U87 glioblastoma multiforme (GBM) tumors in nude mice. Repeated systemic treatment with gemcitabine was used as a positive control. [Modes for carrying out the invention]

[0035] As described above, the present invention provides methods and sustained-release anticancer compositions for local treatment of cancer, prevention of cancer recurrence and inhibition of tumor metastasis.

[0036] In one aspect of the present invention, a method for treating a solid tumor is provided, comprising administering an effective amount of a pharmaceutical composition containing a microparticle biodegradable substrate coated with a polymer / lipid-based matrix containing taxen to a subject having a solid tumor, wherein the pharmaceutical composition is administered directly to the tumor wall of the resected tumor cavity after surgical removal of the tumor. Alternatively, the pharmaceutical composition may be injected directly into the tumor (e.g., an unresected tumor or residual tumor after resection). The method of the present invention is further useful for reducing tumor cell regrowth at the solid tumor resection site after tumor resection surgery. According to certain embodiments, the method of the present invention is useful for treating brain tumors (e.g., glioblastoma multiforme). According to some embodiments, the sustained-release taxen composition is intended to be applied as a single dose during tumor resection surgery or before surgical wound suturing.

[0037] As used herein, “solid tumor” (or “solid carcinoma”) refers to an abnormal mass of tissue that typically does not contain cysts or fluid areas. Solid tumors can be malignant or benign. Malignant solid tumors can invade surrounding tissues and metastasize to new parts of the body. The term “solid tumor” does not include leukemia (cancer that affects the blood). The three main types of solid tumors are sarcomas, carcinomas, and lymphomas. A “sarcoma” is a cancer that arises from connective or supporting tissue, such as bone or muscle. A “carcinoma” is a cancer that arises from glandular and epithelial cells that form the walls of body tissues. A “lymphoma” is a cancer of lymphatic organs, such as lymph nodes, spleen, and thymus. Examples of solid tumors include sarcomas and carcinomas (glioblastoma multiforme, head and neck cancer, colon cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, lung cancer, small cell lung cancer, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endosarcoma, lymphangiosarcoma, lymphangiosarcoma, synovial tumor, mesothelioma, pancreatic cancer, esophageal cancer, gastric cancer, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary carcinoma) Examples include, but are not limited to, adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchial carcinoma, renal cell carcinoma, hepatocellular carcinoma, cholangiocarcinoma, choriocarcinoma, seminomas, embryonal carcinoma, Wilms' tumor, cervical carcinoma, testicular carcinoma, bladder carcinoma, epithelial carcinoma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pineal glandoma, hemangioblastoma, acoustic neuroma, oligodendroneuria, cutaneous T-cell lymphoma (CTCL), melanoma, neuroblastoma, and retinoblastoma.

[0038] According to several embodiments, the methods of the present invention are useful for treating brain tumors and for reducing the regrowth of brain tumor cells at the tumor resection site after surgical removal of brain tumors. Typical examples of brain tumors that can be treated using the compositions and methods described herein include: Gliomas (such as undifferentiated astrocytoma, glioblastoma multiforme, pilocytic astrocytoma, oligodendroglioma, ependymoma, myxopapillary ependymoma, subependymoma, choroid plexus papilloma, etc.); neurotumors (e.g., neuroblastoma, gangliblastoma, ganglioma, and medulloblastoma); pineal gland tumors (e.g., pineoblastoma and pineocytoma); meningeal tumors (e.g., meningioma, intracranial hemangiopercytoma, meningiosarcoma); tumors of nerve sheath cells (e.g., schwannoma (neurolemmoma and neurofibroma)); lymphomas (e.g., Hodgkin lymphoma and non-Hodgkin lymphoma (including numerous subtypes, both primary and secondary)); malformative tumors Tumors (e.g., craniopharyngiomas, epidermoid cysts, dermoid cysts, and colloid cysts); and metastatic brain tumors (which can originate from virtually any tumor, but most commonly from tumors of the lungs, breast, melanoma, kidneys, and gastrointestinal tract).

[0039] In this specification, the terms “treatment” or “to treat” refer to an approach to obtain a beneficial or desired outcome, which includes, but is not limited to, therapeutic benefits and / or preventive benefits. Therapeutic benefits mean at least one of the following: (a) reducing tumor size; (b) inhibiting or reducing tumor growth; (c) reducing or limiting the occurrence and / or spread of metastases; (d) increasing survival or progression-free survival; and (e) delaying the time from tumor removal surgery to tumor recurrence.

[0040] In some embodiments, treating a solid tumor includes inhibiting tumor metastasis. "Inhibiting tumor cell metastasis" can include any degree of inhibition compared to no treatment.

[0041] The term “tumor resection” or “tumor excision” refers to a surgical procedure whose purpose is to remove the entire tumor or as much of it as possible. Some tumors can be easily removed, while others may be located in hard-to-reach places. Typically, a surgeon removes a tumor, but in doing so, removes some of the surrounding normal, healthy tissue (i.e., the “resection margin”) to increase the success rate of the surgery. Those skilled in the art will understand that surgical removal or excision of the entire tumor is not always successful. As used herein, “tumor excision” refers to a state in which at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the tumor volume has been removed by surgery.

[0042] In this specification, the term "tumor resection cavity" refers to the postoperative defect following surgical resection of a tumor. Since complete removal of the tumor is not always possible through surgery, it is understood that the tumor resection cavity may contain residual tumor mass.

[0043] In this specification, the terms “effective dose” or “therapeutic effective dose” refer to the amount of the pharmaceutical composition described herein that is sufficiently effective for the intended use (including, but not limited to, cancer treatment as defined above). In some embodiments, the “effective dose” does not exceed the maximum tolerable dose of taxen used (defined as the maximum amount of free drug that does not cause unacceptable side effects when administered systemically). In some preferred embodiments, the “effective dose” in the method of the present invention is lower than the maximum tolerable dose of taxen. As will be understood by those skilled in the art, the maximum tolerable dose is based on the tolerable systemic toxicity of the drug. However, because systemic exposure to the drug administered locally is significantly lower than exposure to the drug administered systemically, the tolerable dose, as defined for local delivery, may be significantly higher than the maximum tolerable dose in systemic treatment. This is particularly true when the drug is released locally without causing a burst effect. According to some embodiments of the present invention, when the taxane in the pharmaceutical composition is docetaxel, the total amount of docetaxel administered to an adult weighing 60 kg in treatment by the method of the present invention shall not exceed 600 mg, or exceed 500 mg, 450 mg, 400 mg, 350 mg, 300 mg, 290 mg, 280 mg, 270 mg, 260 mg, 250 mg, 240 mg, 230 mg, 220 mg, 210 mg, 200 mg, 190 mg, 180 mg, 170 mg, 160 mg, 155 mg, 150 mg, 145 mg, 140 mg, 135 mg, 130 mg, 125 mg, 120 mg, 115 mg, 110 mg, or 100 mg. Each possibility represents a separate embodiment of the present invention.According to a specific embodiment, the total dose of docetaxel administered in treatment by the method of the present invention is 20-600 mg, or 20-550 mg; 20-500 mg, 20-450 mg, 20-400 mg, 20-350 mg, 20-300 mg, 20-280 mg, 20-260 mg, 20-240 mg, 20-220 mg, 20-200 mg, 20-190 mg, 20-180 mg, 20-170 mg, 20-160 mg, 20-150 mg, 20-140 mg, 20-130 mg, 20-120 mg, 20-110 mg, 20-100 mg, 50-600 mg, or 50-550 mg. These ranges are 50-500mg, 50-450mg, 50-400mg, 50-350mg, 50-300mg, 50-280mg, 50-260mg, 50-240mg, 50-220mg, 50-200mg, 50-190mg, 50-180mg, 50-175mg, 50-170mg, 50-165mg, 50-160mg, 60-160mg, 65-160mg, 70-160mg, 75-160mg, 80-160mg, 85-160mg, 90-160mg, 95-160mg, 100-160mg, 80-150mg, 80-140mg, 80-130mg, and 80-120mg. Each possibility represents a distinct embodiment of the present invention.

[0044] According to some embodiments of the present invention, when the taxane in the pharmaceutical composition is paclitaxel, the total amount of paclitaxel administered to an adult weighing 60 kg in treatment by the method of the present invention shall not exceed 800 mg, or 750 mg, 700 mg, 650 mg, 600 mg, 550 mg, 500 mg, 450 mg, 420 mg, 400 mg, 380 mg, 360 mg, 3 The amounts must not exceed 40 mg, 320 mg, 300 mg, 280 mg, 260 mg, 250 mg, 240 mg, 230 mg, 220 mg, 210 mg, 200 mg, 190 mg, 180 mg, 175 mg, 170 mg, 165 mg, 160 mg, 155 mg, 150 mg, 145 mg, 140 mg, 135 mg, 130 mg, 125 mg, 120 mg, 115 mg, 110 mg, or 100 mg. Each possibility represents a distinct embodiment of the present invention. According to a specific embodiment, the total dose of paclitaxel administered in treatment by the method of the present invention is 60-800 mg, or 60-750 mg, 60-700 mg, 60-650 mg, 60-600 mg, 60-550 mg, 60-500 mg, 60-450 mg, 60-400 mg, 60-350 mg, 60-320 mg, 60-300 mg, 60-295 mg, 60-290 mg, 60-285 mg, 60-280 mg, 60-275 mg, 60-270 mg, 60-265 mg, 60-260 mg, 60-250 mg, 60-240 mg, 60-230 mg, These ranges are 60-220mg, 60-210mg, 60-200mg, 60-190mg, 60-185mg, 60-180mg, 60-175mg, 60-170mg, 60-165mg, 60-160mg, 60-155mg, 60-150mg, 80-300mg, 90-300mg, 100-300mg, 110-300mg, 120-300mg, 130-300mg, 140-300mg, 150-300mg, 160-300mg, 170-300mg, 180-300mg, 190-300mg, 200-300mg, 200-290mg, and 200-280mg. Each possibility represents a distinct embodiment of the present invention.

[0045] According to some embodiments of the present invention, when the taxane in the pharmaceutical composition is cabazitaxel, the total amount of cabazitaxel administered in treatment by the method of the present invention shall not exceed 60 mg, or exceed 80 mg, 75 mg, 70 mg, 65 mg, 60 mg, 55 mg, 50 mg, 45 mg, 42 mg, 40 mg, 38 mg, 37 mg, 36 mg, 35 mg, 34 mg, 33 mg, 32 mg, 31 mg, 30 mg, 29 mg, 28 mg, 27 mg, 26 mg, 25 mg, 24 mg, 23 mg, 22 mg, 21 mg, or 20 mg. Each possibility represents a separate embodiment of the present invention. According to a particular embodiment, the total dose of cabazitaxel administered in treatment by the method of the present invention is 10-80 mg, or 10-75 mg, 10-70 mg, 10-65 mg, 10-60 mg, 10-55 mg, 10-50 mg, 10-45 mg, 10-42 mg, 10-40 mg, 10-38 mg, 10-35 mg, 20-50 mg, 20-45 mg, 20-42 mg, 20-40 mg, 20-38 mg, 20-35 mg, 25-50 mg, 25-45 mg, 25-40 mg, 30-50 mg, 30-45 mg, or 30-40 mg. Each possibility represents a distinct embodiment of the present invention. The term "controlled release" refers to the control of the rate and / or amount of the taxane delivered by the pharmaceutical composition of the present invention. The term "sustained release" means that a pharmaceutically active substance is released over a long period of time.

[0046] The pharmaceutical compositions disclosed herein consist of a particulate biodegradable substrate coated or embedded with a matrix composition comprising: (a) a biodegradable polymer; (b) a lipid component comprising at least one phospholipid having a fatty acid moiety of at least 12 carbon atoms; and (c) a taxane chemotherapeutic agent. In some embodiments, the matrix may further comprise sterols. The matrix composition provides sustained release of pharmaceutically active substances at tumor sites or tumor resection sites in the body of a subject requiring it.

[0047] In certain embodiments, the polymer and the lipids or lipids form a substantially water-free, structurally organized lipid-saturated matrix composition. In some embodiments, the matrix composition has a highly ordered multilayer structure in which the polymer and lipids are structured in the form of multiple alternating layers. In some embodiments, the matrix contains at least about 50% by weight of total lipids.

[0048] According to several embodiments, the pharmaceutical composition of the present invention comprises about 80-93% (w / w) of particulate biodegradable substrate and 7-20% (w / w) of matrix composition based on the total weight of the pharmaceutical composition. According to another embodiment, the particulate biodegradable substrate comprises about 80-92% (w / w), 80-91% (w / w), 80-90% (w / w), 80-89% (w / w), 80-88% (w / w), 80-87% (w / w), 80-86% (w / w), 80-85% (w / w), and 8% of the total weight of the pharmaceutical composition. 1~93%(w / w), 82~93%(w / w), 83~93%(w / w), 84~93%(w / w), 85~93%(w / w), 85~92%(w / w), 85~91%(w / w), 85~90%(w / w), 85~89%(w / w), 85~88%(w / w), 86~89%(w / w).

[0049] In some embodiments, the matrix composition contains at least 10% by weight of the biodegradable polymer. In some embodiments, the matrix composition contains about 10-30% by weight of the polymer. In some embodiments, the matrix composition contains about 15-25% by weight of the polymer. In some embodiments, the matrix composition contains about 20% by weight of the polymer. In some embodiments, the biocompatible polymer accounts for at least 10% (w / w), at least 11% (w / w), at least 12% (w / w), at least 13% (w / w), at least 14% (w / w), at least 15% (w / w), at least 16% (w / w), at least 17% (w / w), at least 18% (w / w), at least 19% (w / w), at least 20% (w / w), at least 21% (w / w), at least 22% (w / w), at least 23% (w / w), at least 24% (w / w), at least 25% (w / w), at least 26% (w / w), at least 27% (w / w), at least 28% (w / w), at least 29% (w / w), and at least 30% (w / w) of the weight of the matrix composition.

[0050] In certain embodiments of the present invention, the polymer is a biodegradable polyester. In some embodiments, the polyester is selected from the group consisting of PLA (polylactic acid). "PLA" refers to poly(L-lactide), poly(D-lactide), and poly(DL-lactide). In another embodiment, the polymer is PGA (polyglycolic acid). In another embodiment, the polymer is PLGA (poly(lactic acid-glycolic acid)). The PLA contained in the PLGA may be any PLA known in the art, for example, an optical isomer or a racemic mixture. In another embodiment, the lactic acid / glycolic acid ratio of the PLGA in the methods and compositions of the present invention is 50:50. In another embodiment, the ratio is 60:40. In another embodiment, the ratio is 75:25. In another embodiment, the ratio is 85:15. In another embodiment, the ratio is 90:10. In another embodiment, the ratio is 95:5. In another embodiment, the ratio is a different ratio suitable for a sustained-release or sustained-release profile in vivo. The PLGA may be either a random copolymer or a block copolymer. Each possibility represents a distinct embodiment of the present invention. Note that the polymer may be of any size or length (i.e., any molecular weight).

[0051] In another embodiment, if the biodegradable polyester includes a hydrogen bond acceptor moiety, the polyester may be selected from the group consisting of polycaprolactone, polyhydroxyalkanoate, polypropylene fumarate, polyorthoester, polyanhydride, and polyalkylcyanoacrylate. In yet another embodiment, the biodegradable polyester is a block copolymer comprising a combination of any two monomers selected from the group consisting of PLA, PGA, PLGA, polycaprolactone, polyhydroxyalkanoate, polypropylene fumarate, polyorthoester, polyanhydride, and polyalkylcyanoacrylate. In yet another embodiment, the biodegradable polyester is a random copolymer comprising a combination of any two monomers listed above. Each possibility represents a distinct embodiment of the present invention.

[0052] The term "biodegradable" refers to a substance that is broken down over time in the human body by hydrolysis, enzymatic action, and / or other similar mechanisms. "Biodegradable" further includes the breakdown or reduction of a substance into non-toxic components within the body after or during the release of a therapeutic agent.

[0053] According to some embodiments, the matrix composition contains a lipid component comprising at least about 30% (w / w of the total weight of the matrix composition) of a lipid component comprising at least one phospholipid having at least 12 carbon fatty acid moieties. According to some embodiments, the matrix composition contains at least about 40% (w / w) of a lipid component comprising at least one phospholipid having at least 12 carbon, preferably 12 to 18 carbon fatty acid moieties, preferably where the hydrocarbon chain is fully saturated. According to some embodiments, the matrix composition contains about 40 to 75% (w / w) of a lipid component comprising at least one phospholipid having at least 12 carbon fatty acid moieties. According to some embodiments, the matrix composition contains about 50 to 70% (w / w) of a lipid component comprising at least one phospholipid having at least 12 carbon fatty acid moieties. According to a particular typical embodiment, the matrix composition contains about 60% (w / w) of a lipid component comprising at least one phospholipid having at least 12 carbon fatty acid moieties. In some embodiments, a lipid component comprising at least one phospholipid having a fatty acid moiety of at least 12 carbon atoms accounts for at least 40% (w / w), at least 45% (w / w), at least 50% (w / w), at least 55% (w / w), at least 60% (w / w), at least 65% (w / w), or at least 70% (w / w) of the total weight of the matrix composition. In some embodiments, a lipid component comprising at least one phospholipid having a fatty acid moiety of at least 12 carbon atoms accounts for 75% (w / w) or less, 70% (w / w) or less, or 65% (w / w) or less of the total weight of the matrix composition. According to some embodiments, the lipid component comprises at least one phospholipid molecule having a fatty acid moiety of at least 14 carbon atoms. According to some embodiments, a second lipid component comprises at least one phosphatidylcholine molecule having a fatty acid moiety of at least 14 carbon atoms. According to some preferred embodiments, the phosphatidylcholine molecule of the composition comprises DMPC. According to several embodiments, the phosphatidylcholine molecule of the composition comprises DPPC.In some embodiments, the phosphatidylcholine molecule of the composition comprises DSPC. In some embodiments, the matrix composition comprises DOPC. In some embodiments, the matrix composition comprises a mixture of DMPC and a second phospholipid having a fatty acid moiety of at least 14 carbon atoms. In some embodiments, the matrix composition comprises a mixture of DMPC and DPPC. Typically, the ratio of DMPC to DPPC in the matrix formulation is about 10:1 to 1:10. In some embodiments, the matrix composition comprises about 50 to 70% (w / w) of DMPC or a DMPC / DPPC mixture.

[0054] In some embodiments, the sustained-release matrix composition may further contain sterols. In some embodiments, the sterols constitute up to 40% (w / w) of the total weight of the matrix composition. In some embodiments, if the sterols are present, they are non-covalently bonded to the biodegradable polymer. In some embodiments, the sterols constitute up to about 30% (w / w) of the total weight of the matrix composition. In some embodiments, the sterols constitute about 5-40% (w / w), about 5-30% (w / w), about 5-20% (w / w), about 5-15% (w / w), about 7-13% (w / w), and about 9-11% (w / w) of the total weight of the matrix composition. In a particular typical embodiment, the matrix composition contains about 10% sterols (w / w of the total weight of the matrix composition). In some embodiments, the sterols constitute at least 5% (w / w), at least 6% (w / w), at least 7% (w / w), at least 8% (w / w), at least 9% (w / w), at least 10% (w / w), at least 11% (w / w), at least 12% (w / w), at least 13% (w / w), at least 14% (w / w), at least 15% (w / w), at least 16% (w / w), at least 17% (w / w), at least 18% (w / w), or at least 19% (w / w) of the matrix. In some embodiments, sterols constitute 20%(w / w) or less, 19%(w / w) or less, 18%(w / w) or less, 17%(w / w) or less, 16%(w / w) or less, 15%(w / w) or less, 14%(w / w) or less, 13%(w / w) or less, 12%(w / w) or less, 11%(w / w) or less, 10%(w / w) or less, 9%(w / w) or less, 8%(w / w) or less, 7%(w / w) or less, 6%(w / w) or less, or 5%(w / w) or less of the matrix composition. Each possibility represents a distinct embodiment of the present invention. According to a particular preferred embodiment, the sterols are cholesterol.

[0055] In some embodiments, the lipid:polymer weight ratio in the pharmaceutical composition of the present invention is in the range of 1:1 to 9:1. In another embodiment, the ratio is in the range of 2:1 to 9:1. In yet another embodiment, the ratio is in the range of 3:1 to 9:1. In yet another embodiment, the ratio is in the range of 4:1 to 9:1. In yet another embodiment, the ratio is in the range of 5:1 to 9:1. In yet another embodiment, the ratio is in the range of 6:1 to 9:1. In yet another embodiment, the ratio is in the range of 7:1 to 9:1. In yet another embodiment, the ratio is in the range of 8:1 to 9:1. In yet another embodiment, the ratio is in the range of 1.5:1 to 9:1. Each possibility represents a distinct embodiment of the present invention.

[0056] It should be noted that the sustained release period using the compositions of the present invention can be programmed taking into account the biochemical and / or biophysical properties of the polymers and lipids. Specifically, the degradation rate of the polymers and the fluidity of the lipids must be taken into consideration. For example, PLGA(85:15) polymer degrades more slowly than PLGA(50:50) polymer. At body temperature, phosphatidylcholine(12:0) is more fluid (less rigid and more amorphous) than phosphatidylcholine(18:0). Therefore, for example, the release rate of a drug incorporated into a matrix composition containing PLGA(85:15) and phosphatidylcholine(18:0) is slower than that of a drug incorporated into a matrix composed of PLGA(50:50) and phosphatidylcholine(14:0). Another aspect that determines the release rate is the physical properties of the encapsulated or impregnated drug. In addition, the drug release rate can be further controlled by adding other lipids to the matrix formulation (some of which will be discussed later).

[0057] In various embodiments, the taxane chemotherapeutic agent encapsulated in the matrix composition coating the microparticle substrate may be any suitable taxane, including, but not limited to, paclitaxel, docetaxel, cabazitaxel, taxadiene, baccatin II, taxuskinin A, brevifoliol, taxispin D, combinations thereof, or pharmaceutically acceptable salts thereof. In various embodiments, the taxane is docetaxel. In various embodiments, the taxane is paclitaxel. In some embodiments, the taxane accounts for about 3-20% (w / w) of the total weight of the matrix composition. According to several embodiments, the taxane is present in amounts of approximately 3-19% (w / w), 3-18% (w / w), 3-17% (w / w), 3-16% (w / w), 3-15% (w / w), 3-14% (w / w), 3-13% (w / w), 3-12% (w / w), 3-11% (w / w), 3-10% (w / w), 3-9% (w / w), 3-8% (w / w), 4-15% (w / w), 4-14% (w / w), 4-13% (w / w), 4-12% (w / w), and 4- 11%(w / w), 4~10%(w / w), 4~9%(w / w), 4~8%(w / w), 5~15%(w / w), 5~14%(w / w), 5~13%(w / w), 5~12%(w / w), 5~11%(w / w), 5~10%(w / w), 5~ 9%(w / w), 5~8%(w / w), 6~15%(w / w), 6~14%(w / w), 6~13%(w / w), 6~12%(w / w), 6~11%(w / w), 6~10%(w / w), 6~9%(w / w), 6~8%(w / w). According to a particular embodiment, the taxane accounts for approximately 0.2% to 2.6% (w / w) of the total weight of the pharmaceutical composition.Alternatively, approximately 0.3-2.5%, 0.3-2.4%, 0.3-2.3%, 0.3-2.2%, 0.3-2.1%, 0.3-2.0%, 0.3-1.9%, 0.3-1.8%, 0.3-1.7%, 0.3-1.6%, 0.3-1.5%, 0.3-1.4%, 0.3-1.3%, 0.3-1.2%, 0.3-1.1%, 0.3-1.0% of the total weight of the pharmaceutical composition. 0.3~0.0%, 0.5~2.5%, 0.5~2.4%, 0.5~2.3%, 0.5~2.2%, 0.5~2.1%, 0.5~2.0%, 0.5~1.9%, 0.5~1.8%, 0.5~1.7%, 0.5~1.6%, 0.5~1.5%, 0.5~1.4%, 0.5~1.3%, 0.5~1.2%, 0.5~1.1%, 0.5~1.0%, 0.6~2.5%, 0.6~2.4%, 0.6~2.3%, 0.6~2.2%, 0.6~2.1%, 0.6~2.0%, 0.6~1.9%, 0.6~1.8%, 0.6~1.7%, 0.6~1.6%, 0.6~1.5%, 0.6~1.4%, 0.6~1.3%, 0.6~1.2%, 0.6~1.1%, 0.6~1.0%, 0.6~0.9%, 0.7~2.5%, 0.7~2.4%, The percentages are 0.7-2.3%, 0.7-2.2%, 0.7-2.1%, 0.7-2.0%, 0.7-1.9%, 0.7-1.8%, 0.7-1.7%, 0.7-1.6%, 0.7-1.5%, 0.7-1.4%, 0.7-1.3%, 0.7-1.2%, 0.7-1.1%, 0.7-1.0%, 0.7-0.9%, 0.8-1.0%, and 0.8-0.9% (w / w). Each possibility represents a distinct embodiment of the present invention. According to some embodiments, the taxane is paclitaxel. According to some embodiments, the taxane is docetaxel.

[0058] In some embodiments, the particulate biodegradable substrate used in the pharmaceutical compositions and methods of the present invention typically consists of spherical or steroidal particles. In some embodiments, the particles do not necessarily have to be spherical and / or steroidal, but are preferably spherical and / or spherical, and their average diameter (as measured by laser diffraction using, for example, Malvern's Mastersizer 3000 instrument) may be at least about 30 μm, at least about 40 μm, at least about 50 μm, at least about 60 μm, at least about 70 μm, at least about 80 μm, at least about 90 μm, at least about 100 μm, 30 μm to 120 μm, 30 μm to 100 μm, 50 μm to 100 μm, about 150 μm or less, about 140 μm or less, about 130 μm or less, about 120 μm or less, about 110 μm or less, or about 100 μm or less. Each possibility represents a separate embodiment of the present invention. According to several embodiments, the particulate substrate used in the compositions and methods of the present invention is a bioabsorbable hydrophilic material that is biocompatible (i.e., low toxicity, exhibits only low levels of foreign body reactions in living organisms, and may have good affinity to body tissues), bioabsorbable (i.e., biodegradable), and hydrophilic, but has low solubility in water; due to this low solubility, the period required for complete disappearance or dissolution in the body is 4 weeks or more, 6 weeks or more, 8 weeks or more, and preferably 10 weeks or more, and the material has a solid form at ambient temperature and is moldable. Any material having these properties can be used non-limitingly. According to several embodiments, the biodegradable substrate is selected from the group consisting of hydroxyapatite, calcium carbonate hydroxyapatite, α-tricalcium phosphate (α-TCP), β-tricalcium phosphate (β-TCP), amorphous calcium phosphate, tetracalcium phosphate, anhydrous dicalcium phosphate, anhydrous monocalcium phosphate, octacalcium phosphate, disodium monohydrogen phosphate, and other bioceramics and combinations thereof based on phosphates. According to some embodiments, the particulate substrate is composed of tricalcium phosphate (TCP), preferably β-TCP.In other embodiments, the particulate substrate consists of polyvinyl alcohol (PVA), preferably PVA having a degree of hydrolysis of at least 88%. In some embodiments, the biodegradable substrate is a porous substrate having porosities in the ranges of 40-80%, 45-80%, 50-80%, 55-80%, 60-80%, 65-80%, and 65-75%. Each possibility represents a distinct embodiment of the present invention.

[0059] In this specification, the term "average diameter size" means that, as measured by laser diffraction, at least about 50% of the substrate particles have a size smaller than the measured average diameter size. For example, particles with an average particle size of 100 μm mean that at least about 50% of the particles have a diameter of less than 100 μm.

[0060] In certain embodiments, the pharmaceutical composition is substantially water-free. In one embodiment, “substantially water-free” means a pharmaceutical composition containing less than 2% water by weight, based on the total weight of the pharmaceutical composition. In another embodiment, the above term refers to a matrix composition containing less than 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, and 0.5% water by weight, based on the total weight of the pharmaceutical composition. In yet another embodiment, the above term means that there is no amount of water that would affect the water resistance properties of the matrix composition. In yet another embodiment, the above term refers to a pharmaceutical composition prepared without the use of any aqueous solvent. In yet another embodiment, lipid saturation is possible by preparing the pharmaceutical composition using a substantially water-free process as described herein. Lipid saturation imparts in vivo bulk degradation resistance to the matrix composition; therefore, the matrix composition exhibits the ability to mediate sustained release on a scale of several days to several weeks (up to about 10 weeks). The total amount of water in the composition may be determined by methods known in the art, such as the Karl Fischer method and water loss by drying.

[0061] Technical basis of pharmaceutical composition used in the method of the present invention According to several embodiments, a sustained-release matrix composition coating a microparticle biodegradable substrate has a highly ordered multilayer structure in which the polymer forms one layer and the phospholipids form a second type of layer, and these two types of layers are configured in the form of multiple alternating or quasi-alternating layers. According to several embodiments, the matrix composition includes a continuous structure without internal gaps and / or free volume. The coating matrix composition is lipid-saturated, meaning that the spaces between polymer layers or polymer backbones are filled with lipid molecules combined with taxanes to such an extent that the matrix can no longer significantly incorporate any further lipid portions.

[0062] The coating matrix compositions disclosed herein are lipid-saturated. “Lipid-saturated” as used herein means that the polymer of the matrix composition is saturated with lipid components (e.g., phospholipids, and optionally sterols) in combination with taxanes present in the matrix, and any other lipids that may be present. The matrix composition contains and is saturated with lipids. In another embodiment, “lipid-saturated” means that the internal gaps (free volumes) within the lipid matrix are filled, with the outer edge of the polymer backbone as the boundary. These gaps are filled with phosphatidylcholine, and optionally with cholesterol, and optionally in combination with other types of lipids, and with taxanes present in the matrix, but to such an extent that the matrix can no longer significantly incorporate additional lipid portions. The lipid-saturated matrices of the present invention offer the further advantage of not requiring synthetic emulsifiers such as polyvinyl alcohol or surfactants; therefore, the matrix compositions of the present invention are typically substantially polyvinyl alcohol-free.

[0063] In some embodiments, when exposed to and maintained in an aqueous medium, the matrix composition is capable of releasing at least 40% of the taxane in a zero-order rate process. In some embodiments, when maintained in an aqueous medium, at least 50%, at least 55%, and at least 60% of the taxane are released from the matrix composition in a zero-order rate process. Although not limited by any particular theory or mechanism of action, it is suggested that the ordered structure or substructure of the matrix composition of the present invention is the primary cause of the zero-order release rate at which the drug (single or multiple drugs) is released from the hydrated matrix formulation. Thus, this zero-order release rate may be due to the slow and continuous "exfoliation" of the hydrated surface layer of the highly ordered layers of lipids and polymers, in which the taxane as a component of the surface layer detached from the matrix is ​​also released. Since this process slowly repeats itself, it is inferred that the taxane is released at a steady rate over several days or weeks until the matrix is ​​completely decomposed. Even without theoretical basis, it is conceivable that the polymer forms a first type of layer, and the phospholipid forms a second type of layer, and that these layers are arranged alternately, i.e., (polymer)-(phospholipid)-(polymer)-(phospholipid); the term “semi-alternating” as used herein refers to a state in which two or more types of layers are alternately present, e.g., (polymer)-(phospholipid)-(phospholipid)-(polymer)-(phospholipid)-(polymer).

[0064] In some embodiments, the matrix composition has multiple mixed layers of polymers and phospholipids as described above, but not in the form of microspheres, micelles, reverse micelles, and liposomes. In some embodiments, the matrix composition does not contain micelles, reverse micelles, or liposomes.

[0065] In some embodiments, the matrix of the present invention is water-resistant. Therefore, even if water is present, it cannot easily diffuse into the inner layers of the matrix, and even if the taxane agent encapsulated between the inner layers were to diffuse, it could not easily diffuse out of the matrix. More specifically, the composition is largely not exposed to water, or the amount of permeating water is small and insufficient to cause the breakdown or decomposition of the matrix bulk (the majority being, for example, the portion of the composition surrounded by the outer surface exposed to the surrounding environment). Without needing to rely on theory or mechanism of action, the water resistance of the matrix composition, along with its inherent multilayer structure, imparts sustained release properties to the matrix (for example, the ability to release at least 40%, preferably at least 50%, 60%, or at least 70% of the taxane chemotherapeutic agent from the composition at zero-order kinetics over periods ranging from several days to several weeks and even several months, provided the composition is maintained in an aqueous environment at physiological temperature).

[0066] The efficacy of a drug is usually determined by its local concentration. Local concentration, on the other hand, is determined by the ratio between the rate of accumulation of the drug released from the formulation and its physical diffusion into surrounding tissues, as well as its elimination by neutralization and / or degradation. An optimal drug delivery system needs to release the drug according to biological needs, with the objective of creating an effective concentration in the vicinity of or immediately adjacent to the target for a sufficient period of time necessary to obtain the desired biological effect. This can be achieved by releasing the drug at the target site at a rate that yields an effective concentration higher than the minimum effective concentration, and for the desired period of time necessary for an effective therapeutic effect. Surprisingly, pharmaceutical compositions according to several embodiments of the present invention have been found to be able to treat solid tumors even when the total dose of the drug (e.g., docetaxel) administered (encapsulated in the pharmaceutical composition) was less than 30% of the maximum tolerated dose of the drug based on the drug's prescription information, and to prevent local recurrence after tumor resection surgery. Furthermore, similar results were obtained even when the tumor was taxane-resistant.

[0067] One of the advantages of the compositions and methods of the present invention is the ability to control local exposure to the taxane by controlling the rate of taxane delivery to the local site. This delivery rate is determined by (1) the taxane release profile; (2) the release rate; and (3) the release duration. These parameters are closely related; the release rate is highly dependent on the particular formulation, while the duration is a function of two factors: the release rate and the drug reservoir size. The pharmaceutical compositions of the present invention, comprising a specific combination of lipids and polymers containing a taxane, preferably docetaxel, not only determine the release rate profile of the taxane but also enable control over the release rate during the course of a sustained zero-order kinetic phase. Without needing to rely on theory or mechanism of action, it is suggested that the most effective and safe release profile for chemotherapeutic agents is a continuous zero-order kinetic release without an initial burst, over a sufficient duration, for example, up to 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40 days, up to 6 weeks, up to 7 weeks, up to 8 weeks, up to 9 weeks, up to 10 weeks, preferably about 14 to 35 days.

[0068] "Zero-order release rate" or "zero-order release kinetics" refers to the steady, linear, continuous, sustained, and controllable release rate of taxane from the pharmaceutical composition, i.e., the plot of taxane release rate against time is a straight line. According to several embodiments, at least 40%, preferably at least 50%, and more preferably at least 60% of the taxane is released from the composition at rates of about 1-7%, 1-6%, 1-5%, 1-4%, 1-3%, 2-7%, 2-6%, 2-5%, 2-4%, and 2-3% ([weight percentage of taxane released per day] / [total weight of taxane initially encapsulated in the composition]) by zero-order kinetics. Each possibility represents a distinct embodiment of the present invention.

[0069] According to some embodiments, when maintained in an aqueous medium at physiological temperature, 1-10% of the taxane is released from the composition by the end of day 1; 10-50% of the taxane is released from the composition by the end of week 1; 20-100% of the taxane is released from the composition by the end of the first two weeks; and 30-100% of the taxane is released from the composition by the end of the first three weeks. In some embodiments, when maintained in an aqueous medium at physiological temperature, at least 10-50% of the taxane is released by the end of week 1; at least 20-80% of the taxane is released by the end of week 2; and at least 30% of the taxane is released by the end of week 3. At least 40% of the taxane is released by the end of week 3. At least 50% of the taxane is released by the end of week 3. By the end of the third week, at least 60% of the taxane is released. According to the currently preferred embodiment, the taxane is docetaxel.

[0070] The pharmaceutical composition used in the method of the present invention delivers taxanes locally to the tumor site or tumor resection site by predictable, long-term sustained release. Therefore, it is possible to maintain a low or zero systemic level of taxane while maintaining a local level of taxane at the tumor site. This sustained local release of taxanes allows a safe dose of topical taxanes to treat tumors and prevent recurrence very effectively at a lower dose than a single dose typically administered intravenously. For example, on the surface of a tumor resection cavity approximately 5 cm in diameter (approximately 25 cm). 2 The amount of docetaxel in 10 grams of the pharmaceutical composition used in the method of the present invention (where docetaxel accounts for about 0.7-1% of the total weight of the composition), which is suitable for application to the estimated lumen surface, is about 50% of the recommended amount of docetaxel that is usually administered by a single intravenous dose once every three weeks.

[0071] Furthermore, the pharmaceutical composition acts as a reservoir protecting the encapsulated taxane. In contrast to conventional polymer-based delivery systems, this property allows for protection of the susceptible drug reservoir not only from biological degradation factors such as enzymes, but also from in vivo soluble substances and chemical degradation due to hydration. This property is particularly important when sustained efficacy is required.

[0072] treatment The present invention addresses a medical need that is currently a concern in the medical community, as there is no effective solution for treating solid tumors and preventing recurrence after tumor resection surgery. The present invention provides local tumor treatment and tumor recurrence prevention, which can be directly applied to the cavity of the tumor resection site during or immediately after tumor resection surgery, or directly administered as neoadjuvant therapy by intratumor injection. The present invention is suitable for cancer treatment, cancer recurrence prevention, and cancer metastasis prevention in a variety of solid tumors.

[0073] In some embodiments, the present invention provides a method for treating a brain tumor, the method comprising administering a therapeutically effective amount of a pharmaceutical composition to the surface of a solid brain tumor or to the surface of the resection cavity after resection of a solid brain tumor: (a) Particulate biodegradable substrate; (b) Biodegradable polymers; (c) at least one phospholipid having a hydrocarbon chain of at least 12 carbon atoms; and (d) Takisen. In some embodiments, the brain tumor is glioblastoma multiforme. In some embodiments, the pharmaceutical composition further comprises sterols. In various embodiments, the taxene is selected from the group consisting of docetaxel, paclitaxel, derivatives of paclitaxel, and cabazitaxel. In certain embodiments, the taxene is docetaxel. In some embodiments, the biodegradable polymer is polyester. In some embodiments, the biodegradable polymer is PLGA. In some embodiments, the phospholipid is phosphatidylcholine having a hydrocarbon chain of 12 to 18 carbon atoms. In certain embodiments, the phospholipid component comprises DMPC. According to several embodiments, a pharmaceutical composition used in a method for treating brain tumors comprises: (a) 80-93% (w / w) tricalcium phosphate; (b) 1-4.0% (w / w) PLGA; (c) 0.0-2.0% (w / w) cholesterol; (d) 4.0-15.0% (w / w) DMPC; (e) 0.2-2.6% (w / w) docetaxel. According to several embodiments, the docetaxel accounts for 0.5-1.5% (w / w) of the total weight of the pharmaceutical composition. According to certain embodiments, the docetaxel accounts for 0.7-1.3% (w / w) or 0.7-1.0% (w / w) of the total weight of the pharmaceutical composition. According to several embodiments, the tricalcium phosphate (TCP) is selected from the group consisting of α-tricalcium phosphate, β-tricalcium phosphate, and combinations thereof. In certain embodiments, the TCP is β-tricalcium phosphate. In some embodiments, the pH of the pharmaceutical composition is approximately 7.5 to 8.5. In some embodiments, the pharmaceutical composition for treating brain tumors further comprises a pH adjuster. In some embodiments, the pH of the pharmaceutical composition is approximately 4 to 6. In some embodiments, a pH of 4 to 6 stabilizes the taxane (e.g., docetaxel) and reduces its conversion to the 7-epimer. In certain embodiments, a method for treating brain tumors includes topical administration of the pharmaceutical composition disclosed above to the surface of a solid brain tumor or to the surface of the resection cavity after resection of a solid brain tumor.In some embodiments, brain tumor resection as used herein refers to a state in which at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the tumor volume has been surgically removed. If surgical removal of the brain tumor is not possible, and is inoperable, or if the patient with the tumor is unable to undergo surgery due to their medical condition, the pharmaceutical composition may be injected directly into the tumor. In certain embodiments, the pharmaceutical formulation comprises: (a) 85-92% (w / w) tricalcium phosphate; (b) 2.0-3.0% (w / w) PLGA; (c) 0.0-2.0% (w / w) cholesterol; (d) 4.0-10.0% (w / w) DMPC; and (e) 0.5-1.5% (w / w) docetaxel. In some exemplary embodiments, the pharmaceutical composition comprises: (a) 86-89% (w / w) tricalcium phosphate; (b) 2.4-2.8% (w / w) PLGA; (c) 0.8-1.5% (w / w) cholesterol; (d) 7.0-9.0% (w / w) DMPC; and (e) 0.6-1.3% (w / w) docetaxel. In some embodiments, the tricalcium phosphate is β-tricalcium phosphate. The brain tumor treatments disclosed above reduce, minimize, or effectively avoid the delay between surgical tumor removal and the initiation of currently performed adjuvant therapies such as radiotherapy and systemic chemotherapy (typically about four weeks after surgery and only after the surgical wound healing process has begun). In some embodiments, the brain tumor treatments of the present invention further inhibit the formation of tumor metastases.

[0074] In some embodiments, the methods disclosed above are suitable for the treatment of primary brain tumors. Primary brain tumors can arise from different types of brain cells, or from the membranes (meninges), nerves, or glands surrounding the brain. The most common type of primary tumor in the brain is the glioma, which arises from the glial tissue of the brain. In some embodiments, the glioma is an astrocytoma. In some embodiments, the astrocytoma is selected from the group consisting of grade I (pilocytic cell) astrocytoma, grade II (fibrous) astrocytoma, grade III (undifferentiated) astrocytoma, and grade IV glioblastoma multiforme (GBM). In other embodiments, the glioma is an oligodendrocyte. In yet another embodiment, the glioma is an ependymoma. In some embodiments, the brain tumor is a secondary or metastatic brain tumor. Secondary or metastatic brain tumors arise from cancer cells that have migrated from a tumor that originated in another part of the body. The most common brain metastases originate from lung cancer cells, breast cancer cells, melanoma cells, colorectal cancer cells, and kidney cancer cells.

[0075] In some embodiments, the present invention provides a method for treating colon cancer, the method comprising administering a therapeutically effective amount of a pharmaceutical composition to the surface of a solid colon cancer tumor or to the surface of the resection cavity after resection of a solid cancer tumor: (a) Particulate biodegradable substrate; (b) Biodegradable polymers; (c) at least one phospholipid having a hydrocarbon chain of at least 12 carbon atoms; and (d) Takisen. In some embodiments, the pharmaceutical composition further comprises sterols. In various embodiments, the taxene is selected from the group consisting of docetaxel, paclitaxel, derivatives of paclitaxel, and cabazitaxel. In certain embodiments, the taxene is docetaxel. In some embodiments, the biodegradable polymer is polyester. In some embodiments, the biodegradable polymer is PLGA. In some embodiments, the phospholipid is phosphatidylcholine having a hydrocarbon chain of 12 to 18 carbon atoms. In certain embodiments, the phospholipid component comprises DMPC. In some embodiments, a pharmaceutical composition used in a method for treating colon cancer comprises: (a) 80-93% (w / w) tricalcium phosphate; (b) 1-4.0% (w / w) PLGA; (c) 0.0-2.0% (w / w) cholesterol; (d) 4.0-15.0% (w / w) DMPC; (e) 0.2-2.6% (w / w) docetaxel. In some embodiments, the docetaxel accounts for 0.5-1.5% (w / w) of the total weight of the pharmaceutical composition. In certain embodiments, the docetaxel accounts for 0.7-1.3% (w / w) or 0.7-1.0% (w / w) of the total weight of the pharmaceutical composition. In some embodiments, the tricalcium phosphate (TCP) is selected from the group consisting of α-tricalcium phosphate, β-tricalcium phosphate, and combinations thereof. In certain embodiments, the TCP is β-tricalcium phosphate. In some embodiments, the pH of the pharmaceutical composition is approximately 7.5 to 8.5. In some embodiments, the pharmaceutical composition for treating colon cancer further comprises a pH adjuster. In some embodiments, the pH of the pharmaceutical composition is approximately 4 to 6. In some embodiments, a pH of 4 to 6 stabilizes the taxane (e.g., docetaxel) and reduces its conversion to the 7-epimer. In certain embodiments, a method for treating a colon cancer tumor comprises topical administration of the pharmaceutical composition disclosed above to the surface of a solid colon tumor or to the surface of the resection cavity after resection of a colon cancer tumor.In some embodiments, colon cancer tumor resection as used herein refers to a state in which at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% of the tumor volume has been surgically removed. If surgical removal of the tumor is not possible, and is unresectable, or if the patient with the tumor is unable to undergo surgery due to their medical condition, the pharmaceutical composition may be directly injected into the colon tumor. In certain embodiments, the pharmaceutical formulation comprises: (a) 85-92% (w / w) tricalcium phosphate; (b) 2.0-3.0% (w / w) PLGA; (c) 0.0-2.0% (w / w) cholesterol; (d) 4.0-10.0% (w / w) DMPC; and (e) 0.5-1.5% (w / w) docetaxel. In some exemplary embodiments, the pharmaceutical composition comprises: (a) 86-89% (w / w) tricalcium phosphate; (b) 2.4-2.8% (w / w) PLGA; (c) 0.8-1.5% (w / w) cholesterol; (d) 7.0-9.0% (w / w) DMPC; and (e) 0.6-1.3% (w / w) docetaxel. In some embodiments, the tricalcium phosphate is β-tricalcium phosphate. In some embodiments, the colon cancer treatment method of the present invention further inhibits the formation of tumor metastases. In another embodiment, the treatment method disclosed above for treating colon cancer is also suitable for prostate cancer, lung cancer, pancreatic cancer, breast cancer, esophageal cancer, gastric cancer, head and neck cancer, and soft tissue sarcoma.

[0076] According to several embodiments, the present invention provides a method for inhibiting tumor metastasis, wherein the method is applied to subjects having malignant solid tumors: (a) Particulate biodegradable substrate; (b) Biodegradable polymers; (c) at least one phospholipid having a hydrocarbon chain of at least 12 carbon atoms; and (d) Takisen, The method includes administering a pharmaceutical composition containing [a specific substance], thereby inhibiting tumor metastasis. In some embodiments, the pharmaceutical composition further comprises sterols. In various embodiments, the taxene is selected from the group consisting of docetaxel, paclitaxel, derivatives of paclitaxel, and cabazitaxel. In a particular embodiment, the taxene is docetaxel.

[0077] The method of the present invention is even more useful for treating tumor cells that are resistant to conventional chemotherapy. Chemotherapy-resistant tumor cells may be due to: (a) overexpression of drug efflux pumps such as P-glycoprotein; (b) acquired mutations at the drug binding site of tubulin; (c) differential expression of tubulin isotypes; (d) alteration of apoptosis mechanisms; (e) activation of growth factor pathways; or (f) other biochemical changes (Deepak Sampath et al., Clin Cancer Res 2006;12(11):3459-69). The contribution of each of these mechanisms to clinical resistance is unclear, but a correlation has been observed in P-glycoprotein expression levels in some tumor types. Surprisingly, it has been found that the pharmaceutical compositions disclosed herein can effectively kill chemotherapy-resistant tumor cells. In particular, it has been shown that docetaxel sustained-release pharmaceutical compositions such as those disclosed above efficiently kill docetaxel-resistant cancer cells. Without needing to base it on theory or mechanism of action, the combination of local concentration and sustained release suggests that high-concentration sustained exposure to the drug effectively overcomes resistance mechanisms mediated by the efflux (MDR) pump. A non-exclusive list of chemotherapy-resistant tumor cells includes: HCT-8 colorectal cancer cells (IC). 50 Docetaxel-3070nM, IC 50 Paclitaxel (3290nM), GXF-209 gastric cancer cells, UISO BCA-1 breast cancer cells, P02 pancreatic cells, 3LL Lewis lung cancer cells, KB-8-5 (IC) 50 Docetaxel -8.8nM, IC 50 Paclitaxel (70.2 nM), KB-P-15 (IC 50 Docetaxel -17.6nM, IC 50 Paclitaxel (117nM), KB-D-15 (IC 50 Docetaxel -68.2nM, IC50 Paclitaxel (565.5nM), KB-V-1 (IC 50 Docetaxel - 467.5 nM, IC 50 Paclitaxel (3202nM) and KB-PTX / 099 (IC 50 Docetaxel -8.8nM, IC 50 Paclitaxel (74.1 nM) Epidermal cells, DLD-1 (IC) 50 Docetaxel -16.2 nM, IC 50 Paclitaxel (32.8 nM) and HCT-15 (IC) 50 Docetaxel - 54.1 nM, IC 50 Paclitaxel (434.6 nM) Colorectal cancer cells and A549.EpoB40 non-squamous cell lung cancer (IC) 50 Docetaxel -28.5nM, IC 50 Paclitaxel (127.5 nM). According to several embodiments, the method of the present invention may also be suitable for other chemotherapy-resistant tumors in which resistance is due to overexpression of drug efflux pumps.

[0078] Drug efficacy is typically determined by its local concentration in the interstitial fluid surrounding tumor cells. Local concentration, on the other hand, is determined by the ratio of the rate of accumulation of the drug released from the pharmaceutical composition to its elimination (e.g., by physical diffusion into surrounding tissue). While not limited by theory or mechanism of action, the ability of a bioavailable taxen drug to form a sufficiently long-lasting, local high concentration within the tumor or on the internal surface of the resected site after surgical tumor removal is suggested to be a major factor in the ability of the pharmaceutical compositions disclosed herein to effectively kill tumor cells, and even to effectively kill tumor cells resistant to the drug intended for use (i.e., treating docetaxel-resistant tumors with a pharmaceutical composition containing docetaxel). One way to obtain better control over the local effects of taxens (e.g., docetaxel) is: (1) The release profile of taxene released from the pharmaceutical composition; (2) Its release rate; and (3) The duration of its release, One example is controlling it. These parameters are closely related; the release rate is highly dependent on the specific formulation (i.e., the ratio of polymers, lipids, and taxanes), while the duration is a function of two factors: the release rate and the drug reservoir size (which can be achieved, for example, by changing the ratio of tricalcium phosphate particles to organic component amounts). It is well known in the art that increased drug efflux from intracellular compartments via energy-dependent efflux pumps is a natural mechanism in cells. This mechanism is also the cause of chemotherapy resistance. One way to overcome resistant cells is to overwhelm the efflux pumps with high concentrations of the drug over a long period. Therefore, it is suggested that taxanes can kill taxane-resistant tumor cells as long as the concentration of bioavailable taxanes at the tumor site is sufficient and the duration of exposure of tumor cells to the taxanes is appropriate.

[0079] According to some embodiments, the pharmaceutical composition of the present invention is in powder form. According to some embodiments, the powder is substantially water-free. According to other embodiments, the powder is a dry powder. According to some embodiments, the particle size of the powder is determined by the particle size of the biodegradable mineral substrate. The polymer / lipid matrix coating the biodegradable substrate is partially contained within the internal space of the porous biodegradable substrate. According to several embodiments, the average diameter of the polymer-lipid (as measured by laser diffraction) may be at least about 30 μm, at least about 40 μm, at least about 50 μm, at least about 60 μm, at least about 70 μm, at least about 80 μm, at least about 90 μm, at least about 100 μm, 30 μm to 120 μm, 30 μm to 100 μm, 50 μm to 100 μm, about 150 μm or less, about 140 μm or less, about 130 μm or less, about 120 μm or less, about 110 μm or less, or about 100 μm or less. Each possibility represents a separate embodiment of the present invention. According to several embodiments, the powder is spread on the tumor surface, or scattered on the tumor surface, or applied to the inner surface of the resection cavity. According to several embodiments, the powder is applied to a surface area of ​​1 cm² 2 Spread or spray the composition in an amount ranging from 20 mg to 500 mg per square centimeter on the surface of a solid tumor or resection cavity. According to another embodiment, the composition is spread over 1 cm². 2 The dosage should be within the range of 50mg-400mg, 50mg-350mg, 50mg-300mg, 50mg-275mg, 50mg-250mg, 50mg-225mg, 50mg-200mg, 50mg-180mg, 50mg-170mg; 50mg-160mg; 50mg-150mg; 50mg-120mg; 50mg-100mg; 50mg-100mg; 75mg-160mg; 75mg-120mg; or 75mg-100mg per serving.

[0080] According to certain embodiments of the present invention, the pharmaceutical composition is formulated as a paste before application to the tumor site or the tumor wall of the resected tumor cavity after tumor resection. According to some embodiments, the paste is spread on the tumor surface or applied to the internal surface of the resection cavity. Typically, a paste-like structure is obtained by hydrating the particulate pharmaceutical composition with an aqueous solution, such as saline solution (0.9% physiological saline), before application. According to some embodiments, hydration is carried out at a timing of 2 hours or less before application of the obtained paste to the tumor site, preferably up to 1 hour before application of the obtained paste to the tumor site, and more preferably at a timing of 30 minutes or less before application to the tumor site. According to some embodiments, a paste-like structure is obtained when the amounts of aqueous solution (e.g., saline solution) mixed with the pharmaceutical composition are 0.1:1 to 1:1 (w / w); preferably 0.3:1 to 0.6:1 (w / w). According to some embodiments, the aqueous solution added to the dried pharmaceutical composition powder for paste formation as described above does not change the total volume of the hydrated pharmaceutical composition powder, and therefore the total volume remains almost unchanged. According to some embodiments, the paste is spread on the tumor surface or resection cavity surface to form a thin, uniform layer of up to 5 mm thick; or up to 4 mm thick; or up to 3 mm thick; preferably 1 to 3 mm thick.

[0081] In another embodiment, the pharmaceutical compositions disclosed herein may be administered intratumorally as neoadjuvant therapy, typically by injection, typically before surgery. In some embodiments, the pharmaceutical composition may be injected directly into the tumor as a dry powder using an instrument suitable for injecting dry powder (a non-limiting example is disclosed in U.S. Patent No. 8,579,855, but any other medical instrument known in the art suitable for powder delivery may be used). Alternatively, the pharmaceutical composition may be injected as a liquid suspension. The liquid suspension may be injected using standard clinically used syringes, needles, tube insertion systems, and cannulas. The liquid suspension may preferably be prepared such that a minimum amount of continuous liquid phase is added to a pharmaceutical composition powder suitable for preparing an injection suspension. According to several embodiments, an injectable suspension can be obtained when the amounts of continuous liquid phases (e.g., aqueous phases) mixed with the pharmaceutical composition powder are 0.1:1 to 2:1 (w / w), preferably 0.3:1 to 1:1 (w / w), and more preferably 0.3:1 to 0.6:1 (w / w). The volume of the pharmaceutical suspension to be injected does not exceed 50% of the solid tumor volume, and may preferably be less than 45%, 40%, 35%, 30%, 25%, 20%, or 15% of the tumor volume. Each possibility represents a separate embodiment of the present invention. The volume of the suspension may preferably be divided into one or more injections, and may preferably be injected into different parts of the tumor so that the dose spreads over the entire tumor or substantially over the entire tumor volume. Due to the inherent properties of the biodegradable particulate substrate contained in the pharmaceutical composition of the present invention, the composition is radiopaque and observable by standard clinical X-ray fluoroscopy; therefore, the location of the pharmaceutical composition disclosed herein can be monitored during injection and throughout the course of treatment by, for example, ultrasound imaging; magnetic resonance imaging; X-ray transmission imaging; computed tomography; positron emission tomography or isotopic imaging including gamma camera / SPECT; or magnetic or radio wave positioning systems.

[0082] In some embodiments, the injectable suspension may comprise water (e.g., saline solution) and optionally one or more additives selected from the group consisting of buffers, isotonic agents, viscosity modifiers, lubricants, osmotic regulators, and surfactants. For example, the suspension may comprise the pharmaceutical composition particles, water, and a lubricant. In some embodiments, the suspension comprises, or essentially comprises, water, pharmaceutical composition particles suspended in saline solution, and a surfactant. Non-limiting examples of surfactants that may be used include polysorbates (such as polysorbate 20, polysorbate 21, polysorbate 40, polysorbate 60, polysorbate 61, polysorbate 65, polysorbate 80, polysorbate 81, polysorbate 85, and polysorbate 120), lauryl sulfates, acetylated monoglycerides, diacetylated monoglycerides, and poloxamers. The suspension may comprise one or more isotonic agents. Examples of suitable isotonic agents include, but are not limited to, one or more inorganic salts, electrolytes, sodium chloride, potassium chloride, sodium phosphate, potassium phosphate, sodium, potassium sulfate, sodium bicarbonate and potassium bicarbonate, and alkaline earth metal salts (such as alkaline earth metal inorganic salts, e.g., calcium salts and magnesium salts), mannitol, dextrose, glycerin, propylene glycol, and mixtures thereof. The suspension may contain one or more analgesics. Suitable analgesics include cellulose derivatives such as sodium carboxymethylcellulose, hydroxyethylcellulose, hydroxypropyl methylcellulose, and methylcellulose; gelatin, glycerin, polyethylene glycol 300, polyethylene glycol 400, and propylene glycol. The suspension may contain viscosity modifiers that increase or decrease the viscosity of the suspension. Suitable viscosity modifiers include methylcellulose, hydroxypropyl methylcellulose, mannitol, and polyvinylpyrrolidone. The suspension may contain one or more lubricants. Suitable lubricants include natural and synthetic phospholipids (e.g., DMPC) or hyaluronic acid.

[0083] Examples Example 1: Docetaxel sustained-release formulation containing different phospholipids Preparations containing cholesterol or not, and containing different phosphatidylcholines, were prepared. The ratios between the tested preparation components were as follows: TCP:(DMPC, DPPC, DSPC or DOPC):PLGA:DTX was 1000:90:30:10; and TCP:(DMPC, DPPC, DSPC or DOPC):PLGA:CH:DTX was 1000:90:30:15:10.

[0084] These formulations were prepared according to the following exemplary protocol: (a) PLGA (100 mg), CH (50 mg required), docetaxel (33.3 mg), and phosphatidylcholine (300 mg) were added to eight 5 ml volumetric flasks, and then an EA:EtOH mixture was added to dissolve the solids. (b) The mixture was heated to 40°C to 45°C whenever necessary to aid in the dissolution of phospholipids. (c) 1.5 g of β-TCP particles (50-100 μm) were added to each of eight 30 mm Petri dishes, and 2.25 mL of the eight organic solutions prepared in step (a) was added on top of the TCP. (d) The Petri dish was placed on a dry heating block heated to 45°C and left uncovered for about 45 minutes. Then, to allow the solvent to completely evaporate, it was covered and left under vacuum (at room temperature) overnight. (e) All eight formulations were transferred to 20 ml scintillation vials and stored at 4°C while protected from light.

[0085] Docetaxel release 250 mg of each test formulation was placed in a 20 ml vial, and 5 ml of PBS was slowly added to it. The sample was then placed in a 37°C incubator. The PBS medium was collected and analyzed once daily. Next, 5 ml of fresh PBS was added to this vial. The released drug concentration was quantified using HPLC. Release analysis was completed after 13 days. The remaining formulation was dried overnight under vacuum at room temperature. The amount of docetaxel and its 7-epitrium impurities in the formulation residue were quantified.

[0086] As is evident from Figure 1, docetaxel is released more quickly and efficiently from compositions containing DMPC compared to docetaxel release from similar compositions containing phospholipids with longer hydrocarbon chains and higher phase transition temperatures (e.g., DPPC and DSPC). Compositions containing phospholipids with saturated hydrocarbon chains longer than 14 carbon atoms did not reach the potential for complete release within 6 weeks; this 6-week period is typically a limited time window between tumor resection and subsequent adjuvant therapy (including radiation or systemic chemotherapy typically performed as prophylactic treatment after tumor resection). Furthermore, cholesterol-containing compositions were found to better protect the docetaxel reservoir from 7-epimer conversion of docetaxel compared to similar compositions without cholesterol (Figure 2).

[0087] Example 2: Docetaxel sustained-release formulations containing different amounts of DMPC material PLGA (Corbion, Purac 7502); Docetaxel (DTX) (TAPI); DMPC (lipids); TCP (Cam bioceramics, 50-100 μm)

[0088] The ratio of the formulation components, TCP:DMPC:PLGA:DTX, was 1000:(0, 30, 60, 90, 135):30:10, respectively. Based on the total weight of the formulation, DMPC amounted to 0%, 2.8%, 5.5%, 8%, and 11.5% (w / w). The formulation was prepared, and docetaxel release from the formulation was carried out as described in Example 1 above.

[0089] As is clear from Figure 3, the relative content of 7-epi was highest in the DMPC-free formulation, while it was found to be significantly lower in the DMPC-containing formulation.

[0090] Example 3: Docetaxel sustained-release formulation containing a surfactant A formulation containing the surfactant Tween80 was prepared, and a release profile of the formulation was created as described in Example 1 above.

[0091] Formulations containing either DMPC or DPPC as a lipid component, and further containing Tween-80, were prepared. The ratio of the formulation components, TCP:DMPC:PLGA:DTX:Tween-80, was 1000:90:30:10:(0, 15, 45), respectively (Figure 4A). Formulations containing DPPC as a lipid component were also prepared, and the ratio of the formulation components, TCP:DMPC:PLGA:DTX:Tween-80, was 1000:90:30:10:(0, 15, 45, 90), respectively (Figure 4B).

[0092] Figures 4A and 4B show that while the addition of Tween-80 to the sustained-release composition increased the release rate, it affected the overall release profile, which in the presence of Tween-80 was characterized by undesirable burst releases that could cause significant local and systemic toxicity.

[0093] Example 4: Docetaxel sustained-release formulations containing varying amounts of cholesterol Preparations containing different amounts of cholesterol (CH) were prepared.

[0094] The component ratios of the test preparation were as follows: the ratio of TCP:DMPC:PLGA:DTX:CH was 1000:90:30:10:(0, 15, 30); based on the total weight of the preparation, the cholesterol (w / w) content of the preparation was 0%, 1.3%, and 2.6%.

[0095] It was revealed that the conversion of docetaxel to the 7-epimer was reduced in cholesterol-containing preparations (Figure 1). Furthermore, it was found that the addition of cholesterol was effective in protecting docetaxel during storage (see Table 2).

[0096] Figure 5 shows that the higher the cholesterol concentration, the lower the percentage of docetaxel 7-epimer in the formulation. However, because the solubility of cholesterol in the preparation mixture is limited, it is preferable to use cholesterol at a concentration of less than 2.6% w / w of the total weight of the formulation.

[0097] Table 1 lists further formulations containing various TCP / DMPC / PLGA / cholesterol / DTX, comparing formulations with and without cholesterol.

[0098] [Table 1]

[0099] Table 2 summarizes the results of stability assays performed on formulations I to IV listed in Table 1, showing that the presence of cholesterol reduces, and even completely stops, the formation of the 7-epimer of docetaxel in these formulations.

[0100] [Table 2]

[0101] According to embodiments of the present invention, the presence of cholesterol in a sustained-release docetaxel composition chemically stabilizes the docetaxel, resulting in a composition with a 7-epidocetaxel content of 0.5% after storage for 9 weeks (e.g., at room temperature). Particularly preferably, the 7-epidocetaxel content is less than 0.4% (e.g., about 0.35%, about 0.3%, about 0.25%, about 0.20% or less) after 9 weeks of storage at room temperature.

[0102] The term "chemically stable" means that the chemical structure docetaxel is stable when the pharmaceutical composition of the present invention is stored under conventional conditions. Preferably, the content (%) of 7-epidocetaxel is less than 1%, preferably less than 0.5%, after storage at 2-8°C for at least 24 months.

[0103] Example 5: Sustained-release paclitaxel formulation A sustained-release paclitaxel (PTX) composition was prepared as in Example 1 above. The ratio of the test formulation components was as follows: TCP:(DMPC, DPPC, DSPC, or DOPC):PLGA:CH:PTX, which was 1000:90:30:15:10. The release of paclitaxel from the composition was tracked in the manner described in Example 1 above; its zero-order release profile is shown in Figure 6.

[0104] Example 6: Sustained-release docetaxel formulation (PEG) containing polyethylene glycol A formulation containing PEG4000 as a polymer was prepared as described in Example 1. The ratio of formulation components, TCP:DMPC:PEG:cholesterol:docetaxel, was 1000:90:30:15:10.

[0105] Docetaxel release from formulations containing PEG4000 was tracked using dissolution analysis (USP1 dissolution instrument - Sotax AT7 smart, basket, 50 RPM) and compared with docetaxel release from similar formulations containing PLGA as a polymer.

[0106] One g of the formulation was dissolved in PBS (phosphate-buffered saline) containing 0.5% SDS (500 ml of medium in each container). Sampling was performed at 1 hour, 2 hours, 4 hours, 6 hours, and 24 hours.

[0107] As can be seen in Figure 7, the presence of PEG4000 resulted in a burst release of encapsulated docetaxel, with over 90% of the drug being released within 5 hours. In contrast, docetaxel release from formulations containing PLGA showed a significantly prolonged sustained zero-order kinetics, with 90% of the drug being released within 20 hours.

[0108] Example 7: Evaluation of the in vivo antitumor effect of pharmaceutical compositions containing different amounts of docetaxel (DTX) according to several embodiments of the present invention against recurrence in a CT26 cell line syngeneic tumor mouse model. This study was conducted to evaluate the antitumor effect of a sustained-release formulation according to an exemplary embodiment of the present invention against CT26 colon cancer cell line tumors in BALB / c mice (7-8 weeks old and weighing 16-20+ / - grams at the start of the study) using different docetaxel doses.

[0109] Test product Formulation V - PLEX-DTX (TCP:DMPC:PLGA:DTX(w / w)=1000:90:30:30) containing 2.6% docetaxel Formulation VI - PLEX-DTX (TCP:DMPC:PLGA:DTX(w / w)=1000:90:30:15) containing 1.3% docetaxel Formulation I - PLEX-DTX (TCP:DMPC:PLGA:DTX(w / w)=1000:90:30:10) containing 0.88% docetaxel Formulation VII - PLEX-DTX (TCP:DMPC:PLGA:DTX(w / w)=1000:90:30:3) containing 0.27% docetaxel Control: Physiological saline

[0110] Disease induction Docetaxel-resistant cell lines (IC 50 Transplantation of CT-26 subcutaneous tumors (260 nM). IC of docetaxel-resistant cell lines for comparison. 50The values ​​are in the range of several nM. Examples include NSCLC:A549 cells (1.9 nM), CRC:HCT-116 cells (5.4 nM), and epidermal KB-3-1 cells (1.1 nM) [Preclinical Pharmacologic Evaluation of MST-997, an Orally Active Taxane with Superior In vitro and In vivo Efficacy in Paclitaxel- and Docetaxel-Resistant Tumor Models (Clin Cancer Res 2006, 12:3459-69)].

[0111] 500,000 CT-26 cells were subcutaneously injected into the upper right buttock of mice. After 11 days, the tumor had grown to the desired volume (approximately 400 mm²). 3 Since the animals reached a certain stage, they were divided into five groups, and the mice were anesthetized before tumor excision. Groups 1-4 received subcutaneous administration of the test preparation (200 mg) to the tumor bed, with each group receiving a preparation containing different concentrations of docetaxel (2.6%, 1.3%, 0.88%, or 0.27% w / w (Table 3)); group 5 received local administration of saline. The skin incisions were then sutured using sterile sutures. After surgery, the animals were returned to cages to recover and observed. Tumor size, clinical signs, and body weight were monitored for 43 days.

[0112] [Table 3]

[0113] result At the end of the study (day 43), differences in the number of tumor-free animals were observed among the DTX treatment groups. In the maximum docetaxel dose (5.2 mg / mouse), 4 / 8 animals were tumor-free; in group 2 (2.6 mg / mouse), 5 / 9 animals were tumor-free; in group 3 (1.73 mg / mouse), 7 / 9 animals were tumor-free; and in group 4 (0.52 mg / mouse), 3 / 8 animals were tumor-free. There were no tumor-free animals in group 5. Regarding mean tumor volume, the saline treatment group (group 5; 2091 mm) had the highest average tumor volume. 3 Compared to the DTX treatment group (Group 1, Group 2, Group 3, and Group 4, each with 548 mm 3 , 814mm 3 , 218mm 3 and 872mm 3 The size was significantly smaller in Figure 8 (p<0.05). A large within-group standard deviation reflects significant variability in tumor size within the group.

[0114] The survival rates for groups 1, 2, 3, and 4 were 63% (5 / 8), 56% (5 / 9), 90% (8 / 9), and 50% (4 / 8), respectively, while group 5 (untreated) had a survival rate of 0% (0 / 8). In group 1 (2.6% docetaxel), two animals were euthanized due to severe weight loss (day 19); and the tumor volume was 1500 mm³. 3 Because it exceeded [a certain value], one animal was slaughtered (day 43). In group 2 (1.3% docetaxel), the tumor volume was 1500 mm 3 Three animals were euthanized because the tumor volume exceeded the limit (days 22, 31, and 36); and the death of one animal was confirmed (day 36). In group 3 (0.88% docetaxel), the tumor volume was 1500 mm². 3 Because it exceeded the limit, only one animal was euthanized early (day 15). In group 4 (0.27% docetaxel), the tumor volume was 1500 mm 3 Because it exceeded the limit, four animals were euthanized early (on days 10, 12, and 17). In group 5 (untreated), the tumor volume was 1500 mm³. 3Because the threshold was exceeded, all animals were euthanized on day 24. In the saline control group, all animals were euthanized on day 24, but in the docetaxel formulation treatment group according to several embodiments of the present invention, most animals survived until the end of the experiment (day 43).

[0115] body weight To mitigate the influence of tumor weight on the animal's total body weight, a calibration curve plot of actual tumor weight against tumor volume was created based on the resected tumor. This plot allowed for the estimation of tumor weight based on volume, enabling animal weight measurement during follow-up after the trial by subtracting this tumor weight from the actual body weight of the tumor-bearing animal. During this trial, animal body weight was measured three times a week. This body weight was normalized to the animal body weight on the day of tumor resection and the day the procedure began.

[0116] In animals in groups 1 and 2 (2.6% docetaxel and 1.3% docetaxel, respectively), weight loss occurred, with maximum losses of 20% and 9% on day 17. Weight gain was observed in both groups 1 and 2 on day 17; by the end of the study, these animals were 115–116% of their original weight. In animals in group 3 (0.88% docetaxel), there was mild weight loss (approximately 2%) up to two weeks after administration, but weight gain was observed from day 17 onward, reaching 113% of their original weight by the end of the study. In animals in group 4 (0.27% docetaxel) and the untreated group (group 5), weight gain began on day 3 after surgery.

[0117] Consideration The antitumor effects of treatment with various docetaxel formulations (each containing different concentrations of docetaxel) according to several exemplary embodiments of the present invention were shown compared to a saline-treated group. Compared to the saline-treated group, animal survival increased with all formulations. However, symptoms related to docetaxel toxicity were more frequent with docetaxel formulations containing the maximum concentration of docetaxel (1.3% docetaxel (Formulation VI) and 2.6% docetaxel (Formulation V)).

[0118] Interestingly, lower concentrations of docetaxel (0.88% (Formulation I); 1.76 mg / mouse) resulted in minimal weight loss and were concluded to be safer. At this dose, it was also more effective than the lowest docetaxel concentration (0.27% (Formulation VII); 0.54 mg / mouse) in reducing tumor recurrence in mice.

[0119] Example 8: Evaluation of the antitumor effect of the formulation according to the embodiment of the present invention in a syngeneic tumor mouse model. In this experiment, the efficacy of topical treatment with sustained-release formulations according to several embodiments of the present invention was compared with systemic docetaxel treatment. To this end, colon cancer tumors were created subcutaneously in female BALB / c mice (7-8 weeks old at the start of the study, with a body weight of ±16-20 grams) and a desired volume (400-600 mm) was introduced. 3 After reaching a certain volume, the tumor was resected, removing approximately 90% of its volume, and then the test drug was administered. Tumor recurrence rates were tracked and compared with an untreated control group.

[0120] Test design 500,000 CT-26 cells were subcutaneously injected into the upper right buttock of the animals. Approximately 7 days later, the tumor reached the desired volume (400 mm). 3 After reaching a certain age, the animals were divided into five groups, and the mice were anesthetized before tumor resection. In Group 1, Formulation VI containing 1.3% docetaxel (2.6 mg / mouse) was administered to the tumor bed at a dose of 200 mg. In Group 2, Formulation I containing 0.88% docetaxel (1.72 mg / mouse) was administered to the tumor bed at a dose of 200 mg. In Groups 3 and 4, treatment was performed by repeated intravenous injections of docetaxel solution. In Group 3, after an intravenous injection of 20 mg / kg, 10 mg / kg was administered once every four days for five doses. In Group 4, after an intravenous injection of 30 mg / kg, 15 mg / kg was administered once every four days for five doses. Group 5 was a saline treatment control group, and approximately 100 μL of saline was administered locally to the tumor bed. The skin incision was then sutured with sterile sutures. After surgery, the animals were returned to cages to recover and observed. Tumor size, clinical signs, and body weight were monitored over a 39-day period. The complete study design is shown in Table 4.

[0121] [Table 4]

[0122] Experimental method Test results At the end of the study (day 39), in group 1, 5 out of 8 animals were tumor-free. In group 2, 6 out of 8 animals were tumor-free. In group 3 (intravenous docetaxel), 2 out of 8 animals were tumor-free. In group 4 (intravenous docetaxel), 3 out of 8 animals were tumor-free. In group 5 (saline treatment), all animals had tumors.

[0123] After 39 days, the average tumor volume was 563 mm in treatment groups 1-4 (groups 1, 2, 3, and 4, respectively). 3 , 375mm 3 , 955mm 3 and 485mm 3 In Figure 9), the saline control group (1500 mm) 3 It was significantly smaller than (p<0.05). The large within-group standard deviation reflects the large variability in tumor size within the group.

[0124] The survival rates in the groups treated with the sustained-release formulation according to the embodiments of the present invention were 63% (5 / 8) and 75% (6 / 8) for groups 1 and 2, respectively. The survival rates in the docetaxel IV treatment groups were 50% (4 / 8) and 63% (5 / 8) for groups 3 and 4, respectively. In group 5 (saline control), the survival rate was at most 12.5% ​​(1 / 8). In group 1 (formulation VI, 1.3% docetaxel), the tumor volume was 1500 mm³. 3 Because it exceeded this limit, three animals were euthanized prematurely (on days 18, 30, and 37). In group 2 (formulation I, 0.88% docetaxel), the tumor volume was 1500 mm². 3 Because it exceeded this limit, two animals were euthanized prematurely (on days 30 and 34). In group 3 (intravenous docetaxel 10 mg / kg), the tumor volume was 1500 mm². 3Four animals were euthanized prematurely because they exceeded the limit (three on day 10 and one on day 25). In group 4 (intravenous docetaxel 15 mg / kg), one animal was euthanized prematurely (on day 20) due to severe weight loss and poor physical condition, and the tumor volume was 1500 mm³. 3 Two animals were euthanized prematurely because the threshold was exceeded (on days 10 and 34). In the saline control group, the tumor volume was 1500 mm³. 3 Because the number exceeded this limit, eight animals were slaughtered (four on day 10, and one each on days 16, 20, 23, and 37).

[0125] In this study, animal body weight was measured three times a week using the method described in Example 5 above. Body weight loss occurred in animals in groups 1, 2, 3, and 4, with maximum weight loss of 12% (day 16), 8% (day 16), 8% (day 16), and 17% (day 20), respectively. No body weight loss was observed in animals in group 5 (saline control) due to early tumor development that increases mouse body weight. Overall, body weight began to increase on days 18, 20, and 23 (groups 1, 2, 3, and 4, respectively) in the groups treated with the sustained-release formulations disclosed herein and the intravenous docetaxel treatment groups.

[0126] conclusion Both formulations I and VI showed high efficacy in reducing tumor recurrence and increasing overall survival when applied topically. Both formulations demonstrated comparable efficacy. Systemic docetaxel treatment at 15 mg / kg (2.6 mg / total dose per mouse) was less effective in reducing tumor-free survival compared to topical treatment; this demonstrates the superiority of topical treatment. In addition, systemic treatment induced severe systemic toxicity, which was reflected in weight loss in the animals. Despite similar exposure to total docetaxel doses in both groups (approximately 1.7 mg), weight loss was less pronounced in group 2 (formulation I, 0.88% docetaxel).

[0127] Example 9: Evaluation of the antitumor effect of a sustained-release formulation according to an exemplary embodiment of the present invention on an in vivo mouse xenograft tumor model of the U87 GBM cell line. This study was conducted to evaluate the efficacy of different amounts of sustained-release compositions according to several exemplary embodiments of the present invention in terms of their antitumor effects against U87 human GBM cell line tumor xenografts in nude mice.

[0128] Test design Three million U87 cells were subcutaneously injected (SC) into the upper right buttock of mice. Approximately nine days later, the tumor volume was approximately 400 mm². 3 Once the mice reached a certain stage, they were divided into six groups (n=10 / group), anesthetized, and had their tumors excised. Tumor bed size was measured and recorded. Groups 1, 2, and 3 received topical administration of 20, 50, or 100 mg of 0.87% docetaxel formulation II to the tumor bed, respectively. Group 4 received topical administration of 100 mg of formulation II vehicle (additives only, without DTX) to the tumor bed. Group 5 was a saline control, and approximately 100 μL of saline was topically administered to the tumor bed. Group 6 was a positive control, and was treated with gemcitabine (300 mg / kg administered intraperitoneally every 7 days for 4 doses). The skin incision was then sutured with sterile sutures. After surgery, the animals were returned to cages for recovery and observation. Tumor size, clinical signs, and body weight were monitored for 43 days.

[0129] Test results After tumor resection, the area of ​​the tumor bed was measured. The average area of ​​the tumor bed was 134 ± 17 mm². The application of formulation II was calculated based on the tumor bed area of ​​1 cm². 2 The dosage was normalized to the amount per unit. Details of this normalization rate and docetaxel dosage are shown in Table 5.

[0130] [Table 5]

[0131] At the end of the study (day 43), differences were observed in the number of tumor-free animals among the Formulation II treatment groups. In group 1 (100 mg of Formulation II), 2 / 10 animals were tumor-free; in group 2 (50 mg of Formulation II), 1 / 10 animals were tumor-free; and in group 3 (20 mg of Formulation II), 4 / 10 animals were tumor-free. In group 4 (100 mg of Formulation II vehicle) and group 5 (saline control), all animals had tumors. In group 6 (gemcitabine), 2 / 10 animals were tumor-free. At 43 days (Figure 10), the mean tumor volume was 69 mm in all Formulation II treatment groups and gemcitabine treatment groups (groups 1, 2, 3, and 6, respectively). 3 , 456mm 3 , 403mm 3 and 780mm 3 In the group treated with formulation II by vehicle and the group treated with saline solution (in groups 4 and 5, respectively, 1898 mm 3 and 2059mm 3 It was significantly smaller than (p<0.001).

[0132] The survival rates for groups 1, 2, and 3 (administered 100, 50, or 20 mg of Formulation II, respectively) were 60% (6 / 10), 30% (3 / 10), and 50% (5 / 10), respectively. In group 4 (100 mg of Formulation II vehicle), only 10% (1 / 10) survival was recorded. In group 5 (saline control), no surviving animals were recorded on day 31. In group 6 (gemcitabine), the survival rate was 20% (2 / 10). In group 1 (100 mg of Formulation II), four animal deaths were confirmed (one each on days 20 and 33, and two on day 34). In group 2 (50 mg of Formulation II), six animal deaths were confirmed (one each on days 9, 18, 23, 25, 33, and 39). On day 23, the tumor volume was 1500 mm³. 3One animal was euthanized prematurely because it exceeded the limit. In group 3 (20 mg formulation II), deaths were confirmed in 5 animals (one each on days 9, 18, 23, 25, 33, and 39). All of these animals showed a weight loss of approximately 20% the day before death was confirmed, so the cause of death is likely systemic toxicity. In group 4 (100 mg formulation II vehicle), the tumor volume was 1500 mm 3 Nine animals were euthanized prematurely because the tumor volume exceeded the limit (two on day 9, three on day 13, three on day 18, and one on day 25). In group 5 (saline control), two animals died (one on day 13 and one on day 23). The cause of death was unknown. Tumor volume was 1500 mm³. 3 Because the threshold was exceeded, eight animals were euthanized early (3 on day 9, 2 on day 13, 1 on day 17, 1 on day 27, and 1 on day 30). In group 6 (gemcitabine), the death of four animals was confirmed (1 each on days 30 and 41, and 2 on day 34). Tumor volume was 1500 mm 3 Four animals were euthanized because the threshold was exceeded (one on each of days 23, 27, 30, and 33). The cause of death for most of the animals in the treatment groups (groups 1, 2, 3, and 6) was thought to be systemic toxicity (all of these animals showed a weight loss of approximately 20% the day before death was confirmed).

[0133] Animals in groups 1 and 2 receiving Formulation II (100 mg or 50 mg, respectively) experienced weight loss, with maximum mean weight loss of 9% (day 34) and 2% (day 13), respectively. No weight loss was observed in group 3 (20 mg of Formulation II). Animals in group 4 (Vehicle with Formulation V) showed a maximum mean weight loss of 2% (day 6). Animals in group 5 (saline control) showed a maximum mean weight loss of 5% (day 23). Animals in group 6 (gemcitabine) showed a maximum mean weight loss of 13% (day 34). From the point of maximum weight loss, animal weight began to increase in all groups. On day 43, the body weight of animals in the Formulation II treatment groups was 99%, 100.5%, and 105% of the baseline body weight of groups 1, 2, and 3, respectively. In the saline control group (Group 4), the formulation II vehicle group (Group 5), and the gemcitabine group (Group 6), the number of surviving animals was so small that a statistically significant mean could not be calculated.

[0134] conclusion Different amounts of Formulation II (mg / cm³) 2 The antitumor effect of treatment with (reflected in) was shown compared to the saline control and the Formulation II vehicle treatment group. Animal survival increased compared to the saline control group at all Formulation II treatment levels. Formulation II 20 mg or 50 mg (15 or 37 mg / cm³) 2 In the group treated with ), the average tumor volume was 1898 mm² compared to the saline control group. 3 From each, 403mm 3 and 456mm 3 It decreased to 100 mg formulation II / animal (75 mg / cm³). 2 Treatment with ) showed the greatest effect on human GBM tumor recurrence after surgical resection (maximum number of surviving animals and minimum mean total tumor volume (69 mm)). 3 (This will be reflected in...)

[0135] Example 10: Evaluation of the antitumor effect of a sustained-release formulation according to an embodiment of the present invention against syngeneic 9L GBM cell line tumors in the brain of Fischer rats. This study was conducted to evaluate the antitumor effects of different amounts of sustained-release formulations according to several exemplary embodiments of the present invention on animal survival after syngeneic brain tumor induction in Fischer rats.

[0136] Test design This study used 75 animals. As shown in Table 6, the animals were divided into 9 groups. Group 1 was the untreated control. Groups 2 and 3 were the positive controls and were treated with forced oral administration of low-dose (33.5 mg / kg) and high-dose (50 mg / kg) temozolomide (the SOC chemotherapy treatment in GBM patients), respectively. In Group 4 (n=10), the resected site was treated with a vehicle containing Formulation II, using the same dose as the high-dose Formulation II group. In Groups 5-8 (n=10 / group), the resected site was treated with Formulation II at doses of 5 mg, 10 mg, 25 mg, or 50 mg / defect site. At the start of the study, all animals were incised to expose the skull bone, and then a 5 mm diameter hole (defect) was drilled into the skull bone. The dura mater was cut to expose the brain. In each animal, a stereotactic device was used to locate 9L cells (10) at a depth of approximately 1 mm in the brain. 5 Cells (2 μL / animal) were injected. After cell injection, the incision was sutured. The animals were returned to their cages for recovery. Five days after cell injection, treatment (temozolomide or Formulation II) was initiated. For Formulation II / Formulation II vehicle treatment, on day 5, the brain defects in groups 4-8 were reopened, and the test substance was administered over the injection site inside the defect. The animals were returned to their cages for recovery. Survival, clinical signs, body weight, and cognitive behavioral assessments were continuously monitored throughout the course of this study.

[0137] [Table 6]

[0138] Test results All animals died within 5 weeks after treatment. In Group 1, the mean survival was 15.8 ± 1.9 days. In Group 2 (temozolomide 33.5 mg / kg), the mean survival was 18.8 ± 2.7 days. In Group 3 (temozolomide 50 mg / kg), the mean survival was 21.8 ± 3.3 days. In Group 4 (vehicle of Formulation II), the mean survival was 17.9 ± 2.2 days. In Group 5 (Formulation II 50 mg / animal), the mean survival was 22.8 ± 5.8 days. In Group 6 (Formulation II 25 mg / animal), the mean survival was 20.9 ± 6.5 days. In Group 7 (Formulation II 10 mg / animal), the mean survival was 20.4 ± 4.9 days. In Group 8 (Formulation II 5 mg / animal), the mean survival was 20.4 ± 3.2 days.

[0139] Conclusion Improved animal survival was observed at all test doses following intracranial administration of Formulation II 5 days after intracerebral injection of tumor cells. The antitumor effect was enhanced with the amount of Formulation II administered. Overall, the maximum effect was obtained with 50 mg of Formulation II (0.87% docetaxel w / w) per site corresponding to 2.2 mg / cm of Formulation II (2.2 mg / cm of docetaxel) (defect diameter 5 mm, defect area 0.196 cm). 2 of Formulation II (2.2 mg / cm of docetaxel) 2 corresponding site (defect diameter 5 mm, defect area 0.196 cm) 2 )

[0140] Example 11: Evaluation of the Pharmacokinetic (PK) Profile of a Locally Administered Docetaxel (DTX) Sustained Release Composition According to Exemplary Embodiments of the Present Invention in Rats In this study, the PK profiles of docetaxel sustained release compositions according to multiple embodiments of the present invention administered to rats were compared. The systemic PK profile of docetaxel released from the locally administered formulation was compared with the PK profile of intravenously administered docetaxel.

[0141] Animals 30 female Sprague-Dawley rats with a body weight of + / - 200 grams

[0142] Experimental Design This study included three test groups (n=10). After anesthetizing the animals, a 1 cm incision was made in the upper right buttock and the skin was lifted to create a subcutaneous pocket. Creating this pocket slightly damaged the underlying muscle, mimicking the state of subcutaneous tumor graft excision in a rat model. Each animal underwent the procedures detailed in Table 7. In groups 1 and 2, formulations VI and I were administered onto the damaged muscle in the subcutaneous pocket, respectively. The skin was then sutured. In the intravenous treatment group (group 3), the administration procedure was performed once immediately after wound suturing. Blood samples were collected at predetermined time points after administration. Each treatment group was divided into two subgroups (n=5 / subgroup), and samples were collected from each subgroup at different time points. Blood samples were collected 0.5 hours, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, and 24 hours after administration, as well as 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 14 days, 21 days, and 30 days after administration. Clinical signs and animal body weight were monitored throughout the study. The released docetaxel concentration in plasma samples was evaluated by liquid chromatography-tandem mass spectrometry (LC-MS / MS) (lower limit of quantification [LLOQ] = 3 ng / mL). The PK profile of docetaxel was determined using these results.

[0143] [Table 7]

[0144] Test results PK analysis of plasma samples showed that overall exposure to formulations VI and I was longer than that of a single intravenous administration (formulation VI, formulation I, and intravenous administration were T last These were 168, 120, and 72 hours, respectively; Table 6). Docetaxel exposure time correlated with the dose of docetaxel in the sustained-release formulations (VI and I). Higher docetaxel doses (1.3%) resulted in longer plasma exposure than lower docetaxel doses (0.88%). AUC, C max and t 1 / 2 The same trend was observed in the intravenous formulation C maxwas 10-fold higher than the maximum exposure of formulation VI (881 and 80.4 ng / ml in groups 3 and 1, respectively; Table 4). Since the total dosages of docetaxel for formulation I and intravenous injection were comparable (1.76 and 2 mg / animal, respectively), the AUC values of the two groups were also similarly comparable (2351 and 2276 hour*mg / ml, respectively; Table 8). This observation supports the similar trend of weight change in these two groups.

[0145]

Table 8

[0146] Conclusion Comparison of the systemic PK profiles of docetaxel from sustained-release formulations (formulations I and VI) and intravenous docetaxel administration in rats showed differences in total exposure time and peak exposure. For the total exposure duration, the sustained-release formulations (both DTX concentrations) were longer than a single intravenous injection. The peak plasma level was higher after intravenous docetaxel administration. These differences are due to the gently sustained release of docetaxel from the sustained-release formulations. In formulations I and VI, the exposure period was extended by the sustained-release docetaxel, but the peak plasma level also decreased, limiting the potential exposure to cytotoxic concentrations. T last increased with the dosage of docetaxel in the sustained-release formulation. A similar relationship was also observed for AUC, C max and t 1 / 2 In this study, a sustained-release formulation according to an exemplary embodiment of the present invention demonstrated releasing docetaxel over a long period while maintaining a systemic exposure (AUC) comparable to that of intravenous treatment while significantly reducing C max

[0147] Example 12: Evaluation of local safety after intracranial (IC) administration of a sustained-release formulation according to an exemplary embodiment of the present invention in SD rats ​This study was conducted in Sprague-Dawley rats to evaluate local and systemic safety after intravenous administration of different amounts of sustained-release formulations according to exemplary embodiments of the present invention.

[0148] Test design The animals were divided into 7 groups (n=20 / group). At the start of the study, the bones of the animal's cranial vault were exposed, and a 5 mm diameter hole (defect) was drilled into the cranial vault bone to expose the brain. Groups 1-3 received Formulation II (50 mg, 25 mg, and 10 mg; corresponding to 0.435, 0.218, and 0.087 mg of docetaxel, respectively; plus an additional 255 mg / cm³). 2 , 127 mg / cm³ 2 and 51 mg / cm³ 2 This corresponds to Formulation II (where the pore (defect) size is 0.5 cm in diameter, the surface area is 0.196 cm²). 2 Based on the following, the calculated dose was administered to the brains of the animals. In groups 4-6, the vehicle of formulation II (docetaxel-free) (50 mg, 25 mg, and 10 mg) was administered to the brains of the animals. Group 7 was used as a sham control. After administration of the test substance, the defect was sealed with bone wax and the incision was sutured. The animals were returned to their cages to recover. Clinical signs, body weight, and cognitive behavior (motor activity, tremor, head tilt, and hair rotation) were continuously assessed throughout the course of the study. At each predetermined time point (1, 4, 8, or 16 weeks), five animals from each group were sacrificed, and necropsy was performed to collect the administration site and vital organs for blinded histopathological evaluation.

[0149] During the course of this study, only one animal died on day 89 (from the 25 mg treatment group). One animal was euthanized early on day 90 due to severe weight loss (from the 50 mg treatment group). Both animals showed mild to moderate behavioral changes several days before early euthanasia or death. Necropsy and histological evaluation were not performed on the animal that died because a long period (approximately 24 hours) had passed since death before the time of death confirmation. Necropsy and histological evaluation of the animal that was euthanized early showed no correlation between the administration of formulation II and the animal's condition, leading to the conclusion that weight loss was not related to the test substance.

[0150] With the exception of one animal that experienced severe weight loss (as described above), all other animals in all groups gained weight during the course of this study.

[0151] Histopathological analysis of the skulls and brains of animals sacrificed one week after administration of Formulation II showed that inflammation (1.4–2.4) and necrosis (1.2–3.2) with similar mean grades were present in all animals across all groups in the skull and cortex. No difference in mean scores was observed between different doses of Formulation II and the vehicle for Formulation II (docetaxel-free) in the treatment groups.

[0152] Four weeks after administration, mean necrosis and inflammation scores for the skull and cortex decreased in all groups of Siamese and Formulation II vehicles compared to the Week 1 score. In animals treated with 50 mg and 25 mg Formulation II, mean necrosis and inflammation scores generally increased at Week 4 compared to the end of Week 1. Scores in the 10 mg Formulation II group remained constant between Week 1 and Week 4.

[0153] At the end of week 8, the mean scores for cranial and cortical necrosis decreased in severity in the groups receiving 25 mg and 50 mg of formulation II compared to the end of week 4. Cortical inflammation scores were mild to moderate. In all other groups, inflammation and necrosis scores were zero to minimal.

[0154] At the end of week 16, all formulation II treatment groups had the lowest mean scores for necrosis and inflammation, with the exception of the 25 mg treatment group, which had minimal to mild cranial necrosis scores. The sham group and the formulation II vehicle treatment group had zero necrosis scores and the lowest inflammation scores.

[0155] conclusion Administration of Formulation II did not induce any visible systemic adverse effects. The total dose of docetaxel administered in Formulation II (i.e., up to 50 mg of Formulation II, equivalent to 1–2 mg / kg of docetaxel) was lower than the reported maximum tolerable dose (MTD) and non-lethal dose (NLD) of Taxotere (10 mg / kg intravenous injection); NDA020449) and docetaxel (NDA205924).

[0156] Local release of cytotoxic drugs resulted in local adverse effects, but these effects disappeared over time. In this study, rats received a maximum total dose of 50 mg / 19.6 mm. 2 This supports the safety of administering Formulation II up to this point.

[0157] Example 13: Evaluation of the antitumor and antimetastatic effects of a sustained-release formulation according to an exemplary embodiment of the present invention against an LLC1 cell line in vivo mouse syngeneic tumor model. The objective of this study was to evaluate the antitumor and antimetastatic effects of different doses of formulation II against tumors of a mouse syngeneic Lewis lung cancer (LLC1) cell line in C57BL mice. The selected cell line (LLC1) is known to spontaneously metastasize from primary tumors to the lungs.

[0158] For the above purpose, subcutaneous colon cancer tumors were created in female BALB / c mice (7-8 weeks old at the start of the experiment, with a body weight of ±16-20 grams) to achieve the desired volume (400-600 mm). 3 After reaching a certain volume, the tumor was resected, removing approximately 90% of its volume, and then the test drug was administered. Tumor recurrence rates were tracked and compared with an untreated control group.

[0159] Male C57BL mice weighing 18-21 grams at 7-8 weeks of age were used in this study. LLC1 tumor cells were injected subcutaneously into the back of the mice. The tumors were approximately 400 mm. 3 After reaching a certain volume, excision was performed (at least 90% of the tumor volume was removed; average area 0.7 cm²). 2 The animals were divided into 6 groups (n=10). Details of the study design are listed in Table 9. Groups 1-4 received different amounts of formulation II directly into the tumor bed. The untreated group (group 5) was used as the negative control, and the systemically treated group (group 6) was used as the positive control. Five animals were designated as the Siamese group and did not receive tumor cell injections but underwent surgical treatment (group 7). After the procedure, the surgical site was sutured and the animals were returned to their cages for recovery. 1500mm 3 Animals with tumors exceeding a certain number were euthanized. After completion, the number of lung metastases was counted in each animal.

[0160] [Table 9]

[0161] Test results In Group 1, only one animal was euthanized early on day 21. The tumor did not reach the maximum volume required for early euthanasia, but the animals were euthanized to check whether metastasis had occurred in the tumor-bearing animals in this group. In Group 2, the death of one animal was confirmed on day 14. Three animals were euthanized early: one on day 18 and two on day 21. The tumor did not reach the maximum volume required for early euthanasia, but one animal was euthanized on day 21 to check whether metastasis had occurred in the tumor-bearing animals in this group. The second animal was euthanized due to the size of its tumor. In Group 3, the deaths of four animals were confirmed (on days 11, 18, and two on day 23). On day 21, four animals were euthanized early due to the size of their tumors. In Group 4, the deaths of six animals were confirmed (on days 14, 16, and three on day 21, and on day 23). Two animals were euthanized on day 23 because the tumor exceeded the maximum volume threshold for early euthanasia. In group 5 (untreated), the death of one animal was confirmed on day 16. Due to tumor size, three animals were euthanized on day 14, two on day 16, and one on day 23 (a total of six animals). In group 6, the death of one animal was confirmed on day 25. One animal was euthanized on day 23 because the tumor exceeded the maximum volume threshold for early euthanasia.

[0162] Small changes in mean body weight (%) were recorded across all groups. These changes were generally minimal (approximately 3%) and were mostly observed in Group 5 (untreated) and Group 6 (Taxel); mean body weight at the end was 6.5% and 4% lighter, respectively, than body weight at t=0.

[0163] In Group 1, 6 out of 10 animals had an average tumor volume of 150 mm². 3 The animals had tumors. In group 2, 8 out of 10 animals had an average tumor volume of 1363 mm². 3 They had tumors. In group 3, 9 out of 10 had an average tumor volume of 2097 mm². 3 They had tumors. In group 4, 6 out of 10 had an average tumor volume of 1559 mm². 3The patients had tumors. In group 5 (untreated), 7 out of 10 had an average tumor volume of 2463 mm². 3 They had tumors. In group 6, 4 / 10 had an average tumor volume of 490 mm². 3 He had a tumor.

[0164] The number of metastases was counted after euthanasia / death. In some cases, metastasis could not be evaluated due to the condition of the lungs. The number of lung metastases could not be evaluated due to severe decomposition. Small metastases (0.1-0.5 mm) and large metastases (>0.5 mm) were counted separately. If there were many metastases (>100), it was defined as uncountable (TNTC).

[0165] In Group 1, 5 out of 10 animals did not have metastases. Three animals had small (0.1-0.5 mm) metastases (2, 6, and 7 metastases), and in two other animals, the lungs were too decayed to count. The mean lung weight was 198 ± 55 mg. In Group 2, 4 out of 10 animals did not have metastases. 5 animals had metastases. Two animals had small (3 and 5 metastases), one animal had both small (0.1-0.5 mm) and large (>0.5 mm) metastases (11 and 6, respectively), and in two animals, the number of metastases was too high (>100) to count (TNTC). In one animal, the lungs were too decayed to count. The mean lung weight was 252 ± 87 mg. In Group 3, 3 out of 10 animals did not have metastases. Three animals had small metastases (5, 5, and 4 metastases), three animals had both small and large metastases (6, 10, and 22 small; 1, 4, and 4 large, respectively), and one animal had TNTC metastases. The mean lung weight was 323 ± 115 mg. In group 4, 2 out of 10 animals did not have metastases. Five animals had metastases. Three animals had small metastases (4, 7, and 9 metastases), and two animals had TNTC metastases. In three animals, the lungs were too decayed to be countable. The mean lung weight was 587 ± 481 mg. In group 5 (untreated), metastases were observed in all animals. Eight animals had small metastases (variing from 2 to 20), one animal had both small and large metastases (5 and 3, respectively), and one animal had TNTC metastases. The mean lung weight was 330 ± 64 mg. In group 6, 5 out of 10 animals did not have metastases. Four animals had metastases. Two animals had small metastases (3 and 4 metastases), one animal had both small and large metastases (7 and 2, respectively), and one animal had TNTC metastases. In one animal, the lung was too decayed to be countable. The mean lung weight was 226 ± 114 mg.

[0166] conclusion In this study, therapeutic efficacy was evaluated based on tumor volume and the number of metastases in the lung after surgical resection of the primary tumor. The results of this study revealed that administration of a 100 mg dose of Formulation II was effective in preventing tumor recurrence and inhibiting tumor cell migration after surgical resection, and reduced the number of animals with metastases and the total number of metastases in the lung. These results indicate that local treatment of the tumor bed with pharmaceutical compositions according to embodiments of the present invention has advantages in preventing both tumor recurrence and metastasis.

[0167] Example 14: Evaluation of the penetration of taxanes released from pharmaceutical compositions according to several embodiments of the present invention into rat brains. A sustained-release taxane composition (e.g., Formulation II) according to a specific embodiment of the present invention is administered through a 5 mm pore in the right hemisphere of a rat's brain. At different time points, the animals treated with the sustained-release taxane composition are sacrificed, and their brains are removed and analyzed for the presence of taxane. Specifically, the removed brains are cut horizontally and vertically and multiple 2 mm pores are extracted starting from the injection site of Formulation II. 2 Create cubes. Determine the amount of docetaxel in each of the cubic sections using a validated bioanalytical method for docetaxel in rat brain tissue. Determine the percentage of brain tissue exposed to docetaxel, the diameter of the area exposed to the drug, and the average drug concentration within this area.

[0168] methodology While performing steady saline perfusion at the dura mater level, a treffin bur (coronal saw bit) was used to drill a deep 5mm hole (19.6mm) through the central part of the cranial corona on the right hemisphere. 2 A hole is drilled. Great care is taken to avoid damaging the dural membrane. An elevator blade is inserted into the edge of the resection and moved around the circumference of the hole until the bone fragment of the skull that was drilled is lifted and detached. Next, the dura mater is incised to expose the brain. Then, the paste-like formulation II is applied to the surface of the brain.

[0169] Any method disclosed and claimed herein can be constructed and implemented without excessive experimentation by taking into account this disclosure. While the compositions and methods of the present invention are described in preferred embodiments, it will be apparent to those skilled in the art that modifications can be made to the methods, steps, and order of steps described herein without departing from the concept, spirit, and scope of the invention. More specifically, it will be apparent that certain chemically and physiologically relevant agents can be used as substitutes for the agents described herein while achieving identical or similar results. Any such similar substitutes, which will be apparent to those skilled in the art, are all considered to fall within the spirit, scope, and concept of the invention as defined by the supplementary claims.

Claims

1. A pharmaceutical composition for use in the treatment of solid brain tumors, It contains a biodegradable substrate molded into the shape of fine particles, and the biodegradable substrate is (a) Biodegradable polymers; (b) at least one phospholipid having a hydrocarbon chain of at least 12 carbon atoms; and (c) Taxane It is coated or embedded in a matrix composition containing, A pharmaceutical composition wherein the biodegradable substrate consists of tricalcium phosphate.

2. The pharmaceutical composition according to claim 1, wherein the solid brain tumor is selected from primary brain tumors and metastatic brain tumors.

3. The pharmaceutical composition according to claim 2, wherein the brain tumor is a chemotherapy-resistant tumor.

4. The pharmaceutical composition according to claim 2, wherein the brain tumor is a docetaxel-resistant tumor.

5. The pharmaceutical composition according to claim 2, wherein the primary brain tumor is glioblastoma multiforme.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the pharmaceutical composition is administered to the inner surface of the tumor resection cavity.

7. The pharmaceutical composition has a surface area of ​​1 cm². ２ The pharmaceutical composition according to claim 6, applied to the inner surface of a tumor resection cavity in a dose ranging from 20 mg to 260 mg per unit.

8. The pharmaceutical composition according to any one of claims 1 to 5, wherein the taxane is selected from the group consisting of docetaxel, paclitaxel, and cabazitaxel.

9. The pharmaceutical composition according to claim 8, wherein the taxane is docetaxel.

10. The pharmaceutical composition according to any one of claims 1 to 9, wherein the polymer is polyester.

11. The pharmaceutical composition according to claim 10, wherein the polyester is selected from polylactic acid (PLA), polyglycolic acid (PGA), and poly(lactic acid-glycolic acid) (PLGA).

12. The pharmaceutical composition according to any one of claims 1 to 11, wherein the phospholipid is phosphatidylcholine selected from DMPC, DPPC, DSPC, and DOPC.

13. The pharmaceutical composition according to claim 12, wherein the phosphatidylcholine is DMPC.

14. The pharmaceutical composition according to any one of claims 1 to 13, wherein the biodegradable substrate of the fine particles consists of particles having an average particle size of less than 200 μm.

15. The pharmaceutical composition according to any one of claims 1 to 14, wherein the biodegradable substrate of the fine particles accounts for 80 to 93% (w / w) of the total weight of the pharmaceutical composition.

16. The pharmaceutical composition according to any one of claims 1 to 15, wherein the polymer accounts for 0.5 to 5% (w / w) of the total weight of the pharmaceutical composition.

17. The pharmaceutical composition according to any one of claims 1 to 16, wherein at least one phospholipid having a hydrocarbon chain of at least 12 carbon atoms accounts for 4.0 to 15% (w / w) of the total weight of the pharmaceutical composition.

18. The pharmaceutical composition according to any one of claims 1 to 17, wherein the taxane accounts for a maximum of 2.6% (w / w) of the total weight of the pharmaceutical composition.

19. The pharmaceutical composition according to claim 18, wherein the taxane accounts for 0.5 to 1.5% (w / w) of the total weight of the pharmaceutical composition.

20. The pharmaceutical composition according to claim 19, wherein the taxane accounts for 0.6 to 1.3% (w / w) of the total weight of the pharmaceutical composition.

21. The pharmaceutical composition according to any one of claims 1 to 20, further comprising cholesterol.

22. The pharmaceutical composition according to claim 21, wherein the cholesterol accounts for a maximum of 2% (w / w) of the total weight of the pharmaceutical composition.

23. A pharmaceutical composition according to any one of claims 1 to 22, further comprising a pH adjusting agent.

24. The pharmaceutical composition according to claim 23, wherein the pH of the pharmaceutical composition is 4.0 to 6.

0.

25. The pharmaceutical composition according to any one of claims 1 to 24, wherein the taxane penetrates to a distance of at least 0.5 cm away from the surface of the excised tumor.

26. The pharmaceutical composition according to any one of claims 1 to 25, wherein the pharmaceutical composition is administered as a powder.

27. The pharmaceutical composition according to any one of claims 1 to 25, wherein the pharmaceutical composition is formulated as a paste or an injection suspension before application.

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