Liposome composition of intra-articular corticosteroids for pain control

Abstract

A method for treating joint pain in an individual chronically afflicted with osteoarthritis, the method comprising administering to the osteoarthritis individual repeat doses of a liposomal composition comprising an effective amount of an intra-articular corticosteroid or a pharmaceutically acceptable salt thereof, in an intra-articular administration manner. A robust and long-lasting intra-articular corticosteroid treatment efficacy response is achieved for more than one year, without any signs or symptoms of adrenal insufficiency in the individual.

Description

[0001] Cross-reference to related applications

[0002] This case asserts priority to U.S. Provisional Application No. 63 / 547,217, filed November 3, 2023, and U.S. Provisional Application No. 63 / 633,336, filed April 12, 2024, the full text of which is incorporated herein by reference. Technical Field

[0003] This disclosure relates to a method for treating chronic pain or inflammation using a lipid-based delivery system to deliver a therapeutic agent to a patient with osteoarthritis. This disclosure also relates to a dosing regimen for administering a liposome composition to an osteoarthritis patient requiring long-term treatment for joint pain. Background Technology

[0004] Osteoarthritis (OA) is a common degenerative joint disease of the articular cartilage, associated with hypertrophic bone changes. Treatment of OA mainly involves a combination of nonpharmacological therapies including exercise and lifestyle modifications, pharmacological therapies (such as nonsteroidal anti-inflammatory drugs and intra-articular corticosteroids), glucosamine or chondroitin sulfate supplementation, and joint replacement surgery.

[0005] Intra-articular (IA) corticosteroid injections are an existing treatment recommendation for individuals with osteoarthritis (OA) according to the 2019 Osteoarthritis Research Society International (OARSI) guidelines (Bannuru RR et al., Osteoarthritis and Cartilage, 2019; 27 (11): 1578-1589). Although intra-articular corticosteroid injections are known to relieve osteoarthritis-related pain, reduce inflammation, and improve mobility, their average duration of benefit is usually short, lasting only 2 to 4 weeks (Jüni P. et al. Cochrane Database Syst Rev. 2015;(10) Art. No.: CD005328). Given that osteoarthritis is a chronic and usually progressive disease that leads to joint immobility and causes long-term pain, patients almost always require long-term treatment with multiple doses to maintain their quality of life.

[0006] Repeated injections can be performed in the same joint, but data on the safety and duration of effectiveness of this procedure are limited. Reports indicate that repeated injections of corticosteroids (especially long-term injections) may increase the risk of local and systemic side effects, including pericapsular calcification, tendon rupture, skin atrophy / discoloration, steroid arthritis, joint infection, facial flushing, allergic reactions, and hyperglycemia. For example, chronic use of corticosteroids may lead to negative feedback suppression of the hypothalamic-pituitary-adrenal (HPA) axis, which may further lead to secondary adrenal insufficiency. Therefore, it is generally recommended to use intra-articular corticosteroids (IA) no more than once every 3 months; however, this may result in intermittent therapeutic effects between IA injections and worsen arthritis symptoms.

[0007] Although sustained-release, lipid-based formulations of dexamethasone sodium phosphate (DSP) have been used for intra-arterial corticosteroid (IA) injections in the treatment of pain caused by knee osteoarthritis (international publication number WO2020 / 056399 A1), there remains an unmet need to establish robustness of efficacy response to IA corticosteroid injections in long-term treatment while minimizing or reducing side effects, given the potential risks associated with repeated dosing. Summary of the Invention

[0008] According to some embodiments, this disclosure provides a method for treating joint pain in a human individual with chronic osteoarthritis, comprising administering, via intra-articular injection, repeated doses of a liposomal composition to the human individual, each dose comprising an effective amount of an intra-articular corticosteroid (IACS) or a pharmaceutically acceptable salt thereof, and a lipid mixture, wherein the repeated doses of the liposomal composition do not cause signs and / or symptoms of adrenal insufficiency in the human individual.

[0009] In some embodiments, a dosing regimen for the treatment of chronic joint pain is provided, comprising repeated doses of a liposomal composition administered intra-articularly to the joint of a patient with osteoarthritis, each dose comprising an effective amount of an intra-articular mixture of corticosteroids and lipids, wherein the dosing regimen does not induce signs and / or symptoms of adrenal insufficiency in the patient with osteoarthritis.

[0010] In some embodiments, a method for treating joint pain in a patient with long-term osteoarthritis is provided, comprising administering repeated doses of intra-articular corticosteroids to the patient via intra-articular administration, wherein at least one dose of the intra-articular corticosteroid is prepared in liposomal form, or in a liposomal composition containing an intra-articular corticosteroid or a pharmaceutically acceptable mixture of a salt and lipid thereof, and the effective amount of the intra-articular corticosteroid in the liposomal composition is from about 6 mg to about 18 mg.

[0011] In some implementations, the repeated dose can be administered at dosing intervals that are selected from the following groups: 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, and 36 weeks.

[0012] In some embodiments, the repeated intra-articular administration of intra-articular corticosteroids is released during periods selected from the group consisting of: 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, and 80 weeks.

[0013] In some embodiments, the osteoarthritis is graded as Kellgren-Lawrence grade 2 or 3. In some embodiments, the severity of osteoarthritis is assessed based on the presence of multiple osteophytes observed within the joints of an individual, using radiography or MRI of the joints.

[0014] In some embodiments, the human individual is female. In some embodiments, the human individual is between 50 and 65 years of age, or 65 years of age or older. In other embodiments, the human individual has a body mass index (BMI) greater than 30, approximately 30, or not less than 30.

[0015] In some embodiments, the human individual suffers from unilateral or bilateral osteoarthritis. In some embodiments, the joint pain is knee pain.

[0016] In some implementations, the average daily pain (ADP) score of the human individual is between 5 and 9.

[0017] In some embodiments, the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is released at a rate that does not adversely inhibit the hypothalamic-pituitary-adrenal (HPA) axis. In other embodiments, the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is released at a rate that does not adversely inhibit cortisol levels in a human individual. In other embodiments, the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is released at a rate that does not induce adrenal insufficiency in a human individual.

[0018] In some embodiments, the liposome composition comprises: (a) a lipid mixture containing one or more phospholipids; and (b) an intra-articular corticosteroid or a pharmaceutically acceptable salt thereof. An exemplary intra-articular corticosteroid according to this disclosure is dexamethasone sodium phosphate (DSP). An exemplary lipid mixture comprises dioleoylphosphatidylcholine (DOPC) and dioleoylphosphatidylglycerol (DOPG).

[0019] In some embodiments, the liposome composition contains about 6 mg to about 18 mg, or about 12 mg of intra-articular corticosteroids. In other embodiments, the liposome composition contains about 6 mg to about 18 mg, or about 12 mg of intra-articular corticosteroids per milliliter.

[0020] In some embodiments, the individual has knee osteoarthritis assessed as grade 2 or 3 on the Kellgren-Lawrence scale. The effective amount of IACS or a pharmaceutically acceptable salt thereof in the liposome composition is about 12 mg per milliliter. The liposome composition comprises: (a) IACS or a pharmaceutically acceptable salt thereof; and (b) a mixture of DOPC, DOPG, and cholesterol, wherein the molar percentages of DOPC, DOPG, and cholesterol, based on the total molars of the mixture, are 56.25 to 72.5:7.5 to 18.75:10 to 33.

[0021] Based on changes in pain relative to baseline, the sustained-release liposome composition according to some embodiments of this disclosure was used to treat human individuals with osteoarthritis by single or multiple injections, demonstrating its safety and robustness of efficacy response in long-term treatment with IACS (especially Coopman Group B and Group C steroids in the liposome composition according to this disclosure).

[0022] Other objects, advantages and novel features of the present invention will become clearer from the following detailed description taken in conjunction with the accompanying drawings. Attached Figure Description

[0023] Figure 1 is a schematic diagram illustrating the experimental design of the third phase of the study in this disclosure.

[0024] Figure 2 shows the number of patients who completed injection phase 1 and injection phase 2.

[0025] Figure 3 illustrates the change in the mean of the least squares (LS) of the WOMAC pain score from baseline in the TLC599 treatment group and the placebo group during the first injection period.

[0026] Figure 4 illustrates the mean change in WOMAC functional scores relative to baseline for all treatment groups during the first injection period.

[0027] Figure 5 is a chart illustrating the total cumulative weekly emergency medication consumption for all treatment groups from week 1 to week 24.

[0028] Figure 6 illustrates the mean change in weekly ADP values ​​relative to baseline for all treatment groups during the first injection period (from week 0 to week 24). The abbreviation "NRS" stands for Numerical Rating Scale.

[0029] Figure 7 is a graph illustrating the changes in ADP in all treatment groups from baseline to week 52. The first injection was administered at baseline (week 0), and the second injection was administered at week 24. Detailed Implementation

[0030] Unless otherwise specified, the following terms shall be understood to have the following meanings as used above and throughout this disclosure.

[0031] Unless the context clearly indicates otherwise, the singular forms “a” and “the” used herein include the plural form.

[0032] Unless otherwise stated, all numerical values ​​in this document are to be understood as being modified by “about”. Wherein, when referring to measurable values ​​such as amount, duration, etc., the word “about” means a variable that includes ±10%, preferably ±5%, more preferably ±1%, and even more preferably ±0.1% of the specified value; these variables are appropriate for obtaining the required amount of the liposomal drug.

[0033] As used herein, the terms "treating," "treated," and "treatment" include prevention (e.g., preventative medication), mitigation, prevention, or reversal of progressive structural tissue damage that leads to joint pain. The terms "treatment" and "treatments" may also refer to compositions or agents. Throughout this disclosure, treatment refers to methods that reduce, alleviate, suppress, or delay the symptoms or signs of one or more osteoarthritis conditions, or improve joint pain detected by known techniques, or reduce the use of pain control agents. These methods include, but are not limited to, clinical examination, imaging, or analysis of serum or joint aspirates (e.g., rheumatoid factor, erythrocyte sedimentation rate).

[0034] Pain and its symptoms can be assessed using methods known in the art. These methods include, but are not limited to, the 6-point descriptive pain rating scale, the 11-point NPRS, the visual analog scale, average daily pain (ADP), the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), the Wisconsin Brief Pain Questionnaire, the Brief Pain Inventory, the McGill Pain Questionnaire and the short-form, the McGill Pain Questionnaire, and other rating methods including the Patient Overall Assessment (PGA) of pain management methods. Individuals can use self-reporting to determine the degree of pain, for example, using a rating scale from (0) no pain to (10) maximum pain. For example, the ADP measures daily pain intensity using a numeric rating scale (NRS) from 0 to 10. Alternatively, the WOMAC index is a self-administered questionnaire comprising three subscales (each subscale scored from 0 to 4): pain (5 items), stiffness (2 items), and physical function (17 items). Functional magnetic resonance imaging (fMRI) may be used on individuals, if necessary, to identify pain reduction following administration of the liposomal composition disclosed herein. For example, the disclosed method is considered a treatment if an individual experiences a reduction in joint pain of approximately or at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% compared to a pre-treatment individual or a control individual. Treatment includes a single joint injection or multiple joint injections at desired intervals.

[0035] The term "joint pain" refers to a joint disorder or condition involving inflammation and / or pain in one or more joints. As used herein, "joint pain" encompasses all types and subtypes of arthritis with known or unknown etiologies and causes, including but not limited to: rheumatoid arthritis, osteoarthritis, infectious arthritis, psoriatic arthritis, gouty arthritis, and lupus-related arthritis, or localized tissue pain caused by bursitis, tenosynovitis, epicondylitis, synovitis, and / or other conditions.

[0036] The pharmaceutically acceptable salts of intra-articular corticosteroids (IACS) disclosed herein are salts formed by acidic IACS and a base, i.e., base addition salts, such as alkali metal salts and alkaline earth metal salts (e.g., sodium, lithium, potassium, calcium, and magnesium salts), and ammonium salts (e.g., ammonium salts, trimethylammonium salts, diethylammonium salts, and tri-(hydroxymethyl)methylammonium salts). Similarly, acid addition salts, such as mineral acids, organic carboxylic acids, and organic sulfonic acids (e.g., hydrochloric acid, methanesulfonic acid, and maleic acid), can also be provided to basic IACS.

[0037] The term "long-term" treatment or administration regimen as used in this article generally refers to a treatment or administration regimen that continues for at least 52 weeks when a therapeutic agent is administered to a patient to provide its therapeutic effect. This includes administering a therapeutic agent to a patient in a single dose, multiple doses, or repeated doses over a period of at least 52 weeks (or for a duration of at least 52 weeks). In many cases, the duration of long-term treatment or dosing regimens can be extended to 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80 weeks or longer, depending on the patient's need for improvement, cure or recovery from their original disease state.

[0038] The term "HPA axis" refers to the hypothalamic-pituitary-adrenal axis, a crucial endocrine system regulating the body's stress response and cortisol release. Cortisol is the primary adrenal glucocorticoid and plays a central role in regulating glucose metabolism and the body's stress response. Cortisol produced by the adrenal cortex generates negative feedback inhibition between the hypothalamus and pituitary gland. This effect reduces the secretion of corticotropin-releasing hormone (CRH) and vasopressin, and directly reduces the breakdown of proopiomelanocortin (POMC) to produce adrenocorticotropic hormone (ACTH) and β-endorphins. The use of exogenous glucocorticoids is known to suppress the HPA axis. Therefore, chronic use of glucocorticoids for potent anti-inflammatory and pharmacological effects may exacerbate secondary adrenal insufficiency due to long-term exposure (Younes A. (2017). Translational Pediatrics, 17; 6 (4), 269-273).

[0039] The term "adrenal insufficiency" is clinically classified into primary, secondary, and tertiary causes. Primary adrenal insufficiency occurs due to lesions affecting the adrenal glands themselves. Secondary adrenal insufficiency is caused by a decrease in the level of adrenocorticotropic hormone (ACTH) released by the pituitary gland, and tertiary adrenal insufficiency is caused by a decrease in the level of corticotropin-releasing hormone (CRH) released by the hypothalamus. It is known that glucocorticoid therapy may prolong the suppression of ACTH production, causing suppression of the HPA axis through negative feedback, and further leading to secondary adrenal insufficiency. Some clinical manifestations of secondary adrenal insufficiency include pale skin without obvious anemia, amenorrhea, decreased libido and sexual function, sparse axillary and pubic hair, small testes, secondary hypothyroidism, prepubertal growth retardation, delayed puberty, headache, visual symptoms, and diabetic diabetes insipidus (Oelkers W. (1996). The New England journal of medicine, 335 (16), 1206-1212). The diagnosis of adrenal insufficiency is based on clinical features and confirmed by biochemical tests (e.g., insulin resistance test (ITT), metyrapone stimulation test, or ACTH stimulation test). Characteristics of adrenal insufficiency include fever, hypoglycemia, chronic fatigue, anorexia, diarrhea, weight loss, muscle weakness, abdominal pain, nausea, vomiting, low blood pressure that worsens upon standing and causes dizziness or fainting, irritability and depression, craving for salty foods, irregular or absent menstruation, loss of interest in sex, or other underlying clinical signs and symptoms.

[0040] Lipid mixtures and liposome compositions containing said mixtures

[0041] In some embodiments, this disclosure provides a sustained-release liposome composition comprising a lipid mixture and an effective amount of an intra-articular endothelial steroid (IACS) or a pharmaceutically acceptable salt thereof, wherein the lipid mixture comprises one or more phospholipids, and the sustained-release liposome composition contains phospholipids in an amount of about 20 micromoles (μmol) to about 150 μmol per milliliter (mL).

[0042] In some embodiments, the liposome composition described herein and administered in single or repeated doses can maintain IACS release for 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 19.5 months, 20 months, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, and 3 weeks. 2 weeks, 33 weeks, 34 weeks, 35 weeks, 36 weeks, 37 weeks, 38 weeks, 39 weeks, 40 weeks, 41 weeks, 42 weeks, 43 weeks, 44 weeks, 45 weeks, 46 weeks, 47 weeks, 48 ​​weeks, 49 weeks, 50 weeks, 51 weeks, 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, 80 weeks, half a year, one year, or 1.5 years.

[0043] In some embodiments, the liposome compositions disclosed herein exhibit increased efficacy compared to liposome compositions with a phospholipid content exceeding 150 μmol per 1 mL. In another embodiment, the liposome compositions disclosed herein maintain the therapeutic effect of IACS and reduce IACS-related side effects.

[0044] In some embodiments, the total amount of phospholipids is from about 50 μmol to about 140 μmol per 1 mL of liposome composition. In another embodiment, the total amount of phospholipids is from about 45 μmol to about 135 μmol per 1 mL of liposome composition. In another embodiment, the total amount of phospholipids is from about 70 μmol to about 150 μmol per 1 mL of liposome composition. In another embodiment, the total amount of phospholipids is from about 90 μmol to about 150 μmol per 1 mL of liposome composition. In another embodiment, the total amount of phospholipids is from about 50 μmol to about 120 μmol per 1 mL of liposome composition. In another embodiment, the total amount of phospholipids is from about 60 μmol to about 110 μmol per 1 mL of liposome composition.

[0045] In some embodiments, the liposome composition further comprises at least one pharmaceutically acceptable excipient, diluent, solvent, carrier, active ingredient medium, preservative, cryoprotectant, or combination thereof.

[0046] In some embodiments, the liposome composition of this disclosure is prepared by: mixing one or more phospholipids with one or more buffers, in the presence or absence of cholesterol, to form liposomes; freeze-drying the liposomes with one or more building agents to form a cake-like lipid mixture; and rehydrating the lipid mixture cake with a solution containing IACS to form an aqueous suspension.

[0047] In some embodiments, the liposome compositions of this disclosure are prepared by: mixing one or more phospholipids in a solvent, with or without cholesterol; subsequently removing the solvent to form a lipid mixture in powder or film form; and rehydrating the lipid mixture powder or film with a solution containing IACS to form an aqueous suspension. In another embodiment, the liposome compositions of this disclosure are prepared by: mixing one or more phospholipids in a solvent, with or without cholesterol; and then injecting the dissolved lipid solution into an aqueous solution to form liposomes. The size of the liposomes is reduced by filtration through track-etched polycarbonate membranes. The solvent is removed by diafiltration of the buffer solution using a semi-automatic tangential-flow filtration (TFF) system. The dialyzed liposome solution is then freeze-dried into a powder, and the lipid mixture powder or film is rehydrated with a solution containing IACS to form an aqueous suspension.

[0048] In some embodiments, the liposome compositions of this disclosure comprise about 10% to about 50% of lipid-associated IACS, or about 50% to about 90% of non-associated IACS. The term "non-associated form" refers to IACS molecules that can be separated from the phospholipid / cholesterol portion of the liposome composition by gel filtration and are intended to provide immediate release. In other embodiments, the weight ratio of the phospholipid and cholesterol combination to IACS is about 5 to 80:1. In another embodiment, the weight ratio of the phospholipid and cholesterol combination to IACS is about 5 to 40:1. For example, the weight ratio of the phospholipid and cholesterol combination to IACS may be about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80:1.

[0049] In some embodiments, the IACS concentration in the liposome composition of this disclosure is at least or about 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM or 35 mM; or about 10 mM to about 40 mM, about 15 mM to about 40 mM, about 20 mM to about 40 mM, about 15 mM to about 35 mM, about 15 mM to about 30 mM, about 15 mM to about 25 mM, or about 20 mM to about 25 mM.

[0050] In some embodiments, the total amount of liposome composition administered each time is from about 0.5 mL to about 1.5 mL, preferably about 1.0 mL.

[0051] The lipid mixtures in the liposome compositions provided herein refer to phospholipids or mixtures of phospholipids. These lipid mixtures may be in the form of films, cakes, granules, or powders prior to addition to the liposome composition, but are not limited thereto.

[0052] In some embodiments, with or without cholesterol, the phospholipids or mixtures of phospholipids are pre-formed into liposomes before further processing into lipid mixtures.

[0053] In some implementations, with or without cholesterol, the phospholipids or mixtures of phospholipids are not pre-formed into liposomes before further processing into a lipid mixture.

[0054] Liposomes can be nanoscale and comprise a lipid bilayer encapsulating an internal aqueous agent-carrying component. Non-limiting examples of liposomes include small monolayer vesicles (SUVs), large monolayer vesicles (LUVs), multivesicular liposomes (MVLs), and multilayer vesicles (MLVs).

[0055] Lipid mixtures can be prepared from a variety of lipids capable of forming or incorporating into a monolayer or bilayer structure. The lipids used in this disclosure include one or more phospholipids, including but not limited to phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidic acid (PA), phosphatidylinositol (PI), or combinations thereof. In some embodiments, the lipid mixture comprises egg phosphatidylcholine (EPC), egg phosphatidylglycerol (EPG), egg phosphatidylethanolamine (EPE), egg phosphatidylserine (EPS), egg phosphatidic acid (EPA), egg phosphatidylinositol (EPI), soy phosphatidylcholine (SPC), soy phosphatidylglycerol (SPG), soy phosphatidylethanolamine (SPE), soy phosphatidylserine (SPS), soy phosphatidicacid (SPA), and soy phosphatidylinositol (SPC). Phosphatidylinositol (SPI) or combinations thereof. In other embodiments, the lipid mixture comprises dipalmitoylphosphatidylcholine (DPPC), 1,2-dioleoyl-sn-glycerol-3-phosphatidylcholine (1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylglycero (DPPG), dioleoylphosphatidylglycerol (DOPG), dimyristoylphosphatidylglycerol (DMPG), hexadecylphosphocholine (HEPC), hydrogenated soybean phosphatidylcholine (HPC) Phosphatidylcholine (HSPC), distearoylphosphatidylcholine (DSPC), distearoylphosphatidylglycerol (DSPG), dioleoylphosphatidylethanolamine (DOPE), palmitoylstearoylphosphatidylcholine (PSPC), palmitoylstearoylphosphatidylglycerol (PSPG), monooleoylphosphatidylethanolamine (MOPE), 1-palmitoyl-2-oleoyl-sn-glycerol-3- Phosphatidylcholine (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine, POPC), polyethylene glycol distearoylphosphatidylethanolamine (PEG-DSPE), dipalmitoylphosphatidylserine (DPPS), 1,2-dioleoyl-sn-glycero-3-phosphatidylserine (1,2-dioleoyl-sn-glycero-3-phosphatidylserine (DOPS), dimyristoylphosphatidylserine (DMPS), distearoylphosphatidylserine (DSPS), dipalmitoylphosphatidicacid (DPPA), 1,2-dioleoyl-sn-glycero-3-phosphatidic acid (DOPA), dimyristoylphosphatidic acid (DMPA), distearoylphosphatidic acid (DSPA), dipalmitoylphosphatidylinositol (DPPI), 1,2-dioleoyl-sn-glycero-3- Phosphatidylinositol (1,2-dioleoyl-sn-glycero-3-phosphatidylinositol, DOPI), dimyristoylphosphatidylinositol (DMPI), distearoylphosphatidylinositol (DSPI), or combinations thereof.

[0056] In some embodiments, the lipid mixture comprises a first phospholipid and a second phospholipid. In some embodiments, the first phospholipid is selected from the group consisting of: EPC, EPE, SPC, SPE, DPPC, DOPC, DMPC, HEPC, HSPC, DSPC, DOPE, PSPC, MOPE, POPC, and mixtures thereof; and the second phospholipid is selected from the group consisting of: PG, PS, PA, PI, and mixtures thereof. In some embodiments, the first phospholipid is EPC, EPE, SPC, SPE, DPPC, DOPC, DMPC, HEPC, HSPC, DSPC, DOPE, PSPC, MOPE, POPC, or a mixture thereof; and the second phospholipid is selected from the group consisting of: EPG, EPS, EPA, EPI, SPG, SPE, SPS, SPA, SPI, DPPG, DOPG, DMPG, DSPG, PSPG, DPPS, DOPS, DMPS, DSPS, DPPA, DOPA, DMPA, DSPA, DPPI, DOPI, DMPI, DSPI, hydrophilic polymers having long chains of highly hydrated flexible neutral polymers attached to phospholipid molecules, and mixtures thereof.Examples of hydrophilic polymers include, but are not limited to, polyethylene glycol (PEG), methoxy PEG (mPEG), ganglioside GM1, polysialic acid, polylactic acid (also known as polylactic ester), polyglycolic acid (also known as polyglycolide), polylactic-polyglycolic acid, polyvinyl alcohol, polyvinylpyrrolidone, polymethoxazoline, polyethyloxazoline, polyhydroxyethyloxazoline, polyhydroxypropyloxazoline, polyaspartamide, polyhydroxypropyl methacrylamide, polymethacrylamide, polydimethylacrylamide, polyvinylmethyl ether, and polyhydroxyethyl acrylate, with molecular weights from about 2,000 to about 5,000 Daltons. acrylates, derived celluloses (such as hydroxymethyl cellulose or hydroxyethyl cellulose), and synthetic polymers.

[0057] In some embodiments, the lipid mixture further comprises sterols. There are no particular limitations on the sterols used in this disclosure, and examples include cholesterol, phytosterols (sitosterol, stigmasterol, phycosterol, spinach sterol, brassicasterol, etc.), ergosterol, cholesterolanone, cholesterolenone, cloprostenol, cholesterolyl-2'-hydroxyethyl ether, and cholesterolyl-4'-hydroxybutyl ether. The sterol component in the lipid mixture (if present) can be any sterol conventionally used in the preparation of liposomes, lipid vesicles, or lipid particles. In other embodiments, the lipid mixture comprises about 10 mol% to about 33 mol% of cholesterol, about 15 mol% to about 30 mol% of cholesterol, about 18 mol% to about 28 mol% of cholesterol, or about 20 mol% to about 25 mol% of cholesterol.

[0058] In some embodiments, the lipid mixture comprises a first phospholipid, a second phospholipid, and a sterol in a molar percentage of 29.5% to 87%: 3% to 37.5%: 10% to 33%.

[0059] In some embodiments, the first phospholipid is DOPC, POPC, SPC, or EPC, and the second phospholipid is PEG-DSPE or DOPG.

[0060] In some implementations, the first phospholipid is DOPC and the second phospholipid is DOPG.

[0061] In some implementations, the lipid mixture does not contain fatty acids or cationic lipids (i.e., lipids with a net positive charge under physiological pH conditions).

[0062] The liposome compositions prepared in this disclosure can be prepared using conventional techniques for vesicle preparation. These techniques include ether injection (Deamer et al., Acad. Sci. (1978) 308: 250), surfactant method (Brunner et al., Biochim. Biophys. Acta. (1976) 455: 322), freeze-thaw method (Pick et al., Arch. Biochim. Biophys. (1981) 212: 186), reverse evaporation method (Szoka et al., Biochim. Biophys. Acta. (1980) 601: 559-571), ultrasonic treatment method (Huang et al., Biochemistry (1969) 8:344), ethanol injection method (Kremer et al., Biochemistry (1977) 16: 3932), and extrusion method (Hope et al., Biochim. Biophys. Acta. (1985) 812: The methods described above include 55-65), the French press method (Barenholz et al., FEBS Lett. (1979) 99: 210), and the method described in Szoka, F., Jr., et al., Ann. Rev. Biophys. Bioeng. 9: 467 (1980). All of the above processing techniques are fundamental to vesicle formation, and these processes are incorporated herein by reference. After sterilization, the preformed liposomes are aseptically placed in a container and then freeze-dried to form a powder or cake. In embodiments where the lipid mixture comprises preformed liposomes, the liposomes are prepared by solvent injection and then freeze-dried with or without fillers and / or buffers to obtain the lipid mixture. In some embodiments, the lipid mixture comprises one or more volume-expanding agents. In some embodiments, the lipid mixture further comprises one or more buffers.

[0063] Volume-expanding agents include, but are not limited to: polyols or sugar alcohols, such as mannitol, glycerol, sorbitol, dextrose, sucrose and / or trehalose; and amino acids such as histidine or glycine. Mannitol is a preferred volume-expanding agent.

[0064] Buffers include, but are not limited to, sodium phosphate monobasic dihydrate and anhydrous disodium hydrogen phosphate.

[0065] In some embodiments where the lipid mixture comprises lipids that are not pre-formed into liposomes, the lipid mixture may be prepared by dissolving it in a suitable organic solvent (including but not limited to ethanol, methanol, tert-butanol, diethyl ether, and chloroform) and drying it by heating, vacuum evaporation, nitrogen evaporation, freeze drying, or other conventional solvent removal methods.

[0066] Specific embodiments supporting the preparation of lipid mixtures of this disclosure will be described below.

[0067] Intra-articular corticosteroids

[0068] Endothelial steroids (IACS) are the current treatment recommendations for the non-surgical management of osteoarthritis of the knee, hip and multiple joints proposed by the International Osteoarthritis Research Society (OARSI) guidelines (Bannuru RR et al. Osteoarthritis and Cartilage (2019) 27: 1578-1589).

[0069] IACS that can be used in this disclosure include any naturally occurring steroid hormone, synthetic steroid and its derivatives. Examples of IACS, their derivatives or pharmaceutically acceptable salts include, but are not limited to, corticosteroids according to Coopman classification groups B and C (S. Coopman et al., “Identification of cross-reaction patterns in allergic contact dermatitis from topical corticosteroids” Br JDermatol. 1989 Jul; 121 (1):27-34).

[0070] Pharmaceutically acceptable salts according to IACS include non-toxic salts formed from non-toxic inorganic or organic bases. For example, non-toxic salts can be formed from: inorganic bases, such as hydroxides of alkali metals or alkaline earth metals (e.g., potassium, sodium, lithium, calcium, or magnesium); and organic bases, such as amines.

[0071] Pharmaceutically acceptable salts according to IACS also include non-toxic salts formed from non-toxic inorganic or organic acids. Examples of organic and inorganic acids include hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, succinic acid, citric acid, lactic acid, maleic acid, fumaric acid, palmitic acid, cholic acid, prazinic acid, mucoic acid, D-glutamic acid, glutaric acid, glycolic acid, phthalic acid, tartaric acid, lauric acid, stearic acid, salicylic acid, sorbic acid, and benzoic acid.

[0072] In some embodiments, IACS includes, but is not limited to, hydrocortisone acetate, methylprednisolone acetate, dexamethasone sodium acetate, dexamethasone sodium phosphate, betamethasone acetate, prednisolone, triamcinolone acetonide, and triamcinolone hexacetonide, which can be administered in the following doses: about 0.1 mg to about 300 mg, about 0.1 mg to about 100 mg, about 0.1 mg to about 20 mg, about 0.1 mg to about 18 mg, about 1 mg to about 300 mg, about 1 mg to about 100 mg, about 1 mg to about 20 mg, about 1 mg to about 18 mg, about 4 mg to about 300 mg, about 4 mg to about 10 ...100 mg, about 4 mg to about 100 mg, about 4 mg to about 100 mg, about 4 mg to about 100 mg, about 4 mg to about 100 mg, about 4 mg to about 100 mg, about 4 mg to about 1 mg to about 20 mg, about 4 mg to about 18 mg, about 6 mg to about 18 mg, about 6 mg to about 16 mg, about 8 mg to about 16 mg, about 6 mg to about 12 mg.

[0073] In one embodiment, the IACS is dexamethasone sodium phosphate (DSP). DSP is in solution form, and the cake-shaped lipid mixture is rehydrated as the above-mentioned solution containing IACS to obtain the liposome composition of the present disclosure, such that the IACS concentration of the liposome composition is from about 2 mg / mL to about 100 mg / mL, from about 4 mg / mL to about 80 mg / mL, from about 5 mg / mL to about 60 mg / mL, from about 6 mg / mL to about 40 mg / mL, from about 8 mg / mL to about 20 mg / mL, or from about 10 mg / mL to about 16 mg / mL.

[0074] The effective dose of IACS in humans in this disclosure may be higher than the recommended or standard doses known in the art; see, for example, Wernecke, C. et al. Orthop J Sports Med., 3 (5), 2325967115581163 (DOI:10.1177 / 2325967115581163), which is incorporated herein by reference. For instance, while the recommended and tolerable dose of hexamethylenetetramine cortisol as an IACS is 20 mg, the dose of IACS in the compositions and methods of the present invention may be at least 20 mg or higher.

[0075] The dosage of IACS administered also depends on the severity of the condition being treated, the specific prescription, and other clinical factors such as the recipient's weight and general health condition, as well as the severity of side effects.

[0076] In some embodiments, the liposome composition may further comprise a targeting molecule, including but not limited to TNF-α and B cell surface antigens (e.g., CD20). Other antigens may also be used, such as CD19, HER-3, GD2, Gp75, CS1 protein, mesothelin, cMyc, CD22, CD4, CD44, CD45, CD28, CD3, CD123, CD138, CD52, CD56, CD74, CD30, Gp75, CD38, CD33, GD2, VEGF, or TGF. The targeting molecule may be in the form of a lipid-conjugate of an antibody or peptide, which acts as a targeting portion capable of specifically binding to target cells carrying the targeting molecule, thereby delivering IACS to the desired microenvironment to achieve the desired disease-modifying therapy to improve disease progression.

[0077] Administration of liposome compositions

[0078] The liposomal composition can be administered intra-articularly in a single-dose or multiple-dose regimen during the appropriate course of treatment for the condition. The liposomal composition can be conveniently administered at an appropriate frequency, such as once a week, once every two weeks, once every six weeks, once a month, once every two months, at least once every three months, at least once every six months, at least once every nine months, or until the symptoms and signs of the condition (i.e., joint pain) are relieved, improved, or cured.

[0079] In some embodiments, the liposome composition can be administered at multiple doses, selected from the group consisting of: 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, and 36 weeks.

[0080] In one set of embodiments, the liposome composition can be administered according to a multi-dosing regimen, which includes, but is not limited to, at least two joint injections at dosing intervals selected from the following groups: 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, and 36 weeks.

[0081] In some embodiments, the multiple-dose regimen includes administering at least two joint injections at dosing intervals of at least 12 weeks (e.g., 13 weeks to 6 months). In another set of embodiments, provided that no adverse reaction occurs in the individual after exposure to the liposome composition, the above multiple-dose regimen may further include one, two, three, four or more additional joint injections at appropriate dosing intervals after the two joint injections.

[0082] In some embodiments, the multiple-dose dosing regimen includes administering at least two joint injections at dosing intervals of at least 24 weeks (e.g., 25 to 36 weeks or longer). In another set of embodiments, provided that no adverse reaction occurs in the individual after exposure to the liposome composition, the above multiple-dose regimen may further include one, two, three, four or more additional joint injections at suitable dosing intervals after the two joint injections.

[0083] In some embodiments, the dosage of the liposome composition for each joint injection is about 0.5 mL to about 1.5 mL, about 0.6 mL to about 1.2 mL, about 0.8 mL to about 1.2 mL, or about 1.0 mL.

[0084] In one exemplary embodiment, the liposome composition is administered in a single therapeutic dose, wherein the effective amount of intra-articular corticosteroid is about 6 mg to about 18 mg, about 10 mg to about 18 mg, about 12 mg to about 18 mg, about 10 mg to about 15 mg, about 11 mg to about 13 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, or about 15 mg. The lipid mixture comprises a first phospholipid, a second phospholipid, and a sterol in a molar ratio of 29.5% to 87%: 3% to 37.5%: 10% to 33%; and the total amount of phospholipid is about 50 μmol to about 140 μmol, about 45 μmol to about 135 μmol, about 50 μmol to about 120 μmol, or about 60 μmol to about 110 μmol.

[0085] In some embodiments, the liposome composition is administered in multiple doses, wherein the effective amount of intra-articular corticosteroid in each dose is about 6 mg to about 18 mg, about 10 mg to about 18 mg, about 12 mg to about 18 mg, about 10 mg to about 15 mg, about 11 mg to about 13 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, or about 15 mg. The lipid mixture comprises a first phospholipid, a second phospholipid, and a sterol in a molar ratio of 29.5% to 87%: 3% to 37.5%: 10% to 33%; and the total amount of phospholipid is about 50 μmol to about 140 μmol, about 45 μmol to about 135 μmol, about 50 μmol to about 120 μmol, or about 60 μmol to about 110 μmol.

[0086] In some embodiments, during multi-dose treatment, the liposomal composition is administered as a dose independent of the non-liposomal composition, wherein the effective amount of intra-articular corticosteroid in the non-liposomal composition is from about 0.1 mg to about 300 mg.

[0087] Methods for treating joint pain

[0088] One aspect of this disclosure is a method for treating individual joint pain, comprising administering an effective amount of the liposome composition described herein to an individual in need, thereby reducing IACS-induced adverse reactions compared to individuals administered an immediate-release or standard IACS formulation; and / or improving the therapeutic efficacy and release rate of the liposome composition for IACS compared to the efficacy and release rate of liposome compositions with a phospholipid content exceeding 150 μmol per milliliter of liposome composition.

[0089] In some implementations, an individual suffers from arthritis such as osteoarthritis, rheumatoid arthritis, acute gouty arthritis, psoriatic arthritis, reactive arthritis, or arthritis caused by the following conditions: Ehlers-Danlos Syndrome, hemochromatosis, hepatitis, Lyme disease, Sjogren's disease, Hashimoto's thyroiditis, celiac disease, non-celiac gluten sensitivity, inflammatory bowel disease, allergic purpura, hyperimmune globulin D with recurrent fever, sarcoidosis, Whipple's disease, tumor necrosis factor receptor-associated periodic fever syndrome, granulomatous polyangiitis, familial Mediterranean fever, or systemic lupus erythematosus.

[0090] Planar X-ray or MRI of the knee joint can be used to assess patients with knee osteoarthritis. MRI is the best diagnostic tool for assessing bone and soft tissue changes in knee osteoarthritis. The Kellgren-Lawrence classification (KL classification) of osteoarthritis can be assessed based on knee radiographs.

[0091] The Kellgren and Lawrence grading system has been widely used for classifying the severity of osteoarthritis. The following is the original description: Grade 0 (None): No obvious X-ray changes in osteoarthritis; Grade 1 (Suspicious): Suspicious narrowing of the joint space, possibly with osteophyte-like lip deformity; Grade 2 (mild): Obvious osteophytes are visible on posteroanterior weight-bearing radiographs, and the joint space may be narrowed; Grade 3 (Moderate): Moderate multiple osteophytes, significant narrowing of joint spaces, partial sclerosis of bone ends, possibly accompanied by deformities; Grade 4 (Severe): Numerous osteophytes, significantly narrowed joint spaces, severe sclerosis and obvious deformation of bone ends.

[0092] Patients classified as Grade 2 based on the lowest severity of their diagnosis are considered to have osteoarthritis. This classification was proposed by Kellgren and Lawrence in 1957 and was subsequently adopted by the World Health Organization (WHO) in 1961 as the radiological definition of osteoarthritis (OA) for epidemiological research purposes.

[0093] Therapeutic efficacy refers to the ability of IACS to induce a beneficial clinical response in the disease. Efficacy also includes relief of clinical symptoms, such as joint pain, tenderness, transient morning stiffness, and crepitus that cause unstable and impaired physical function. In some embodiments, the efficacy of IACS can be assessed using indicators such as the WOMAC osteoarthritis index, ADP score, and VAS score. In some embodiments, the IACS in the liposome composition described herein is released sustainably and stably without inducing adverse reactions, including but not limited to articular cartilage damage or destruction, such as chondrocyte apoptosis, proteoglycan loss, articular cartilage cysts, articular cartilage degradation, joint injury, suppression of serum cortisol levels, suppression of the hypothalamic-pituitary-adrenal axis, and adrenal insufficiency. Compared to individuals who received IACS in a non-liposome formulation (e.g., without lipid mixtures) as described in this protocol, the incidence of adverse reactions in the individuals described in this protocol may be reduced by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0094] The liposome compositions provided herein can be used in combination with a variety of additional chemical substances, including but not limited to: analgesics (e.g., bupivacaine, ropivacaine, or lidocaine) or hyaluronic acid preparations (e.g., sine hydrochloride). In some embodiments, the liposome compositions of this application can be prepared as a single therapeutic composition with other chemical substances, and administered simultaneously to both the liposome compositions and the additional chemical substances. Alternatively, the liposome compositions of this application and the additional chemical substances can be independent of each other, for example, prepared as separate therapeutic compositions; and the liposome compositions of this application and the additional chemical substances can be administered simultaneously, or administered separately at different times during the treatment regimen, via the same or different routes, in a single dose or multiple doses.

[0095] This disclosure will be further illustrated with reference to the following specific, non-limiting embodiments.

[0096] Example

[0097] The following examples illustrate the preparation and characteristics of specific embodiments of this disclosure.

[0098] Example 1: Preparation of lipid mixtures

[0099] Lipids (including DOPC, DOPG, and cholesterol) were mixed in a molar percentage ratio of 56.25 to 72.5: 7.5 to 18.75: 10 to 33 (e.g., 67.5: 7.5: 25) and dissolved in 99.9% ethanol at approximately 40°C in a flask to form a lipid solution. The lipids were dissolved using a benchtop ultrasonic cleaner.

[0100] The dissolved lipid solution was added to a 1.0 mM sodium phosphate aqueous solution at a flow rate of 100 mL / min using a peristaltic pump to form a preliposome suspension. The preliposome suspension was then extruded through a 0.2 μm polycarbonate membrane 6 to 10 times to obtain a liposome mixture. The average vesicle size of the liposomes was 120 nm to 140 nm (measured using a Malvern ZetaSizer Nano ZS-90 particle size analyzer, Malvern Instruments Ltd, Worcestershire, UK).

[0101] The liposome mixture was dialyzed and concentrated using a tangential flow filtration system (Millipore Corporation, Billerica, MA, USA) equipped with a Millipore Pellicon 2 Mini Ultrafiltration Module Biomax-100C (0.1 m² filtration area), followed by sterilization using a 0.2 μm sterile filter.

[0102] The lipid concentration of the filtered liposome mixture was quantified using a phosphorous assay. The mixture was then prepared with 2% mannitol and sterilized again using a 0.2 μm sterile filter. Finally, the sterilized liposome mixture was freeze-dried to obtain a cake-like lipid mixture.

[0103] Example 2: Preparation of liposome composition

[0104] The liposome composition according to this disclosure is prepared by mixing the lipid mixture described in Example 1 with an aqueous solution of dexamethasone sodium phosphate (DSP; C22H28FNa2O8P; molecular weight: 516.41 g / L). The DSP aqueous solution contains 13.2 mg / mL of DSP and 4 mg / mL of sodium citrate as the DSP composition for subsequent use. Thus, each milliliter of the liposome composition formed (hereinafter also referred to as "liposome DSP") contains approximately 12.0 mg / mL of DSP and approximately 90 μmol to 100 μmol of phospholipids.

[0105] Example 3: A randomized, double-blinded controlled study on the efficacy and safety of liposomal dexamethasone sodium phosphate (DSP) in patients with knee osteoarthritis.

[0106] Liposome DSP was prepared using the methods described above (e.g., Examples 1 and 2). In a phase III, randomized, double-blind, placebo-controlled, and activity-controlled clinical trial, liposome DSP (hereinafter referred to as "TLC599") was intended for local injection to provide sustained pain relief in knee osteoarthritis (OA). This study aimed to confirm the efficacy of TLC599 in providing pain relief to OA patients over 24 weeks as demonstrated in previous studies, and to investigate the benefits of repeated TLC599 injections within one year.

[0107] The current study aimed to evaluate the efficacy and safety of single-dose or repeated-dose TLC599 in patients with Knee Osteoarthritis (KA) grades 2-3 on the KL Scale. A total of 504 patients (506 enrolled, 2 withdrawn) were randomly assigned in a 2:1:1 ratio to three groups. On day 1 (baseline or week 0), patients received either 12 mg TLC599 (TLC599 treatment group), 4 mg DSP (DSP treatment group), or a placebo (saline) injection. At week 24, 203 eligible patients in the TLC599 treatment group who received their first TLC599 injection received a second TLC599 injection. 89 eligible patients in the DSP treatment group who received their first DSP injection received TLC599 injections. 94 eligible patients in the placebo group who received their first placebo injection received a second placebo injection. The study design is as follows: Figure 1 As shown in Figure 2, this provides an overview of the number of patients who completed the study in both injection phase 1 and injection phase 2. Efficacy and safety were assessed during the first injection phase (defined as a period of up to 24 weeks after the first injection) and the second injection phase (defined as a period of up to 28 weeks after the second injection, or up to 52 weeks after baseline).

[0108] For efficacy parameters, Western University and McMaster University Osteoarthritis Index (WOMAC) pain scores, WOMAC functional scores, and daily average pain (ADP) of the knee were collected and evaluated using statistical methods such as repeated measures mixed model (MMRM) and analysis of covariance (ANCOVA). Simultaneously, safety analysis was based on all safety information collected throughout the study period, including adverse events (AEs), clinical laboratory data, vital signs, electrocardiograms (ECG), and knee radiographs.

[0109] The demographic and baseline characteristics of the patients are summarized in Table 1.

[0110] Table 1. Demographic and Baseline Characteristics

[0111] WOMAC = Western University and McMaster University Osteoarthritis Index.

[0112] ADP = Average daily pain.

[0113] For the efficacy endpoints (ANCOVA) used to assess changes in WOMAC pain at week 12 from baseline, the TLC599 treatment group was statistically significantly superior to the placebo group, with a least square mean difference of -0.171 and a p-value of 0.0372 (Table 2). Furthermore, as shown in Figure 3, the TLC599 treatment group was also numerically superior to the placebo group at all time points up to week 24.

[0114] Table 2. Changes in WOMAC pain scores from baseline to week 12

[0115] Table 3 shows the change in WOMAC function scores from baseline for TLC599 versus placebo during the first injection period. Statistically significant treatment differences were observed up to week 12, and TLC599 showed numerically better performance at every follow-up visit up to week 24 (weeks 16, 20, and 24). The change in WOMAC function scores from baseline for all treatment groups during the first injection period is shown in Figure 4.

[0116] Table 3. Changes in WOMAC functional scores from baseline for TLC599 relative to placebo during the first injection - ANCOVA

[0117] The use of rescue medication (e.g., acetaminophen, opioids, or NSAIDs permitted by the study protocol) is another efficacy assessment of potential clinical benefit. In this study, 500 mg acetaminophen tablets were provided as rescue medication and dispensed upon required screening. Patients recorded their acetaminophen rescue medication usage in a diary daily for the preceding 24 hours between screening and study endpoint / early termination. Rescue medication was reviewed at each study visit. The permitted daily dose of acetaminophen was a maximum of 3 grams per day. Acetaminophen was not permitted to be used within 24 hours prior to each study visit. As summarized in Table 4, the cumulative rescue medication consumption analysis by ANCOVA up to week 12 and week 24 showed a statistically significant difference between the TLC599 treatment group and the placebo group, with the TLC599 treatment group consuming less rescue medication overall. This result supports the significant clinical improvement in mean pain relief in the TLC599 treatment group. The cumulative weekly total emergency medications consumed by each treatment group during the first injection period are shown in Figure 5.

[0118] Table 4. Total emergency acetaminophen consumption analysis of TLC599 versus placebo at weeks 12 and 24 – ANCOVA

[0119] For other knee pain-related assessment indicators, including the change in the weekly mean of daily pain (ADP) from baseline at week 12, and the AUC of the change in WOMAC pain from baseline over the time interval from baseline to week 12, the TLC599 treatment group showed statistically significant improvement in OA knee pain compared to the placebo group. Specifically, at all time points during the first injection, the TLC599 treatment group was numerically and statistically superior (p<0.05) to the placebo group, as shown in Figure 6. As summarized in Table 5, statistically significant treatment differences were observed in the change in the weekly mean of ADP from the first injection between TLC599 and placebo at weeks 1, 12, and 24.

[0120] Table 5. Analysis of the weekly mean change from the first injection in TLC599 versus placebo at weeks 1, 12, and 24 – MMRM

[0121] Furthermore, the efficacy of repeated doses of TLC599 in patients with knee osteoarthritis was further evaluated. Eligible patients in the TLC599 treatment group and placebo group received a second blinded injection of the same treatment at week 24, while patients in the DSP treatment group received a blinded injection of TLC599 concurrently (see Figures 1 and 2). For eligible patients receiving a second injection (N=386), the changes in weekly mean ADP scores from baseline during the first and second injection periods are graphically shown in Figure 7. The TLC599 treatment group demonstrated numerically better therapeutic efficacy from week 1 to week 52 compared to the placebo group in patients receiving a second injection. Similar trends were observed in patients receiving a second TLC599 injection during the second injection period as in patients receiving TLC599 after DSP.

[0122] Regarding safety assessment, clinical laboratory tests, vital signs, ECG, physiological tests, and knee radiography results were all normal and did not raise any safety concerns. No significantly abnormal glucose or HbA1c levels were observed throughout the study.

[0123] Chronic use of corticosteroids is known to suppress the HPA axis through negative feedback, leading to adrenal insufficiency. Normal cortisol levels are typically highest in the morning (10 to 20 μg / dL) and lowest around midnight. Therefore, the safety assessment for potential adrenal insufficiency was performed by measuring morning cortisol levels in each group at predetermined time points / visits. ACTH stimulation was performed on any patient with morning serum cortisol less than 10 μg / dL who also exhibited underlying clinical signs and symptoms suggestive of adrenal insufficiency during the study period (e.g., fever, hypoglycemia, chronic fatigue, anorexia, diarrhea, weight loss, muscle weakness, abdominal pain, nausea, vomiting, hypotension that worsens upon standing, irritability and depression, craving for salty foods, irregular or absent menstruation, loss of interest in sexual activity) to determine whether they had adrenal insufficiency. Specifically, the ACTH stimulation test uses a pre-drug serum cortisol sample taken before 9:00 AM as the baseline level, and a post-stimulation cortisol sample is taken approximately 1 hour after the injection of rehydrated 0.25 mg of synthetic adrenocorticotropic hormone (cosyntropin) to assess post-stimulation serum cortisol levels. Patients who do not pass the ACTH stimulation test (post-stimulation cortisol levels ≤18 μg / dL (or ≤497 nmol / L)) are referred to an endocrinologist for further evaluation and treatment.

[0124] According to the study protocol, patients' cortisol levels were measured at week 20 to qualify for a second injection. Four patients (three in the TLC599 group and one in the placebo group) had morning serum cortisol levels of "0" at week 20 due to sample quality issues. All patients received a second injection after confirming their retested cortisol levels were above 10 μg / dL. At week 20, nine patients in all three treatment groups had morning serum cortisol concentrations below 5 μg / dL. The incidence of patients with morning serum cortisol concentrations below 5 μg / dL was similar across the three groups: four out of 252 patients (1.6%) in the TLC599 group; three out of 126 patients (2.4%) in the DSP group; and two out of 126 patients (1.6%) in the placebo group. The incidence of morning serum cortisol concentrations below 5 μg / dL in the TLC599 group was numerically lower than that in the DSP group and comparable to that in the placebo group, suggesting that this represents a background incidence.

[0125] Furthermore, at week 20, 113 patients showed morning serum cortisol levels of 5 to 10 μg / dL. The frequency of these cases in the TLC599 group (52 out of 252 patients; 20.6%) was lower than in the DSP group (32 out of 126 patients; 25.4%) and the placebo group (29 out of 126 patients; 23.0%). Notably, the incidence of morning serum cortisol levels of 5 to 10 μg / dL in the TLC599 group (20.6%) was numerically lower than in the DSP group (25.4%) and the placebo group (23.0%), indicating that the incidence was comparable across the three treatment groups. Importantly, no patients with cortisol levels below 5 μg / dL or between 5 and 10 μg / dL reported any signs and / or symptoms of adrenal insufficiency at week 20.

[0126] For patients who received at least one injection (Table 6) and for patients who received two injections (Table 7), the proportion of patients with low cortisol (morning serum cortisol <5 μg / dL) was presented in the treatment group. The results in Tables 6 and 7 show that the incidence of low cortisol levels in the placebo group ranged from 1.0% to 7.7% during week 1 to week 52 (end of study). This represents a physiologic sporadic variation within the normal range for low cortisol levels. In other words, there is an inherent background incidence of low cortisol levels and it does not indicate any safety concerns.

[0127] Regarding changes in cortisol levels during the first injection period, as shown in Table 6, the TLC599 group (23.6%) showed a greater shift from baseline to low cortisol levels in week 1 compared to the DSP (4 mg) group (0%) or the placebo group (1.7%). However, in week 2, the shifts to low cortisol levels were similar in the TLC599 group (4.7%) and the DSP group (3.5%), while the incidence in the placebo group was 1.7%. In week 4, the shifts to low cortisol levels were similar in the TLC599 group (3.4%), the DSP group (3.5%), and the placebo group (4.5%).

[0128] A similar pattern was observed in injection phase 2, as shown in Table 7. One week after the TLC599 injection at week 24, 28.6% of patients receiving TLC599 for the first time and 31.6% of patients receiving DSP (4 mg) for the first time had low cortisol levels at week 25, compared to 1.2% of patients receiving a second injection of placebo. At week 26 (two weeks after the second injection), 5.2% of patients in the TLC599 group and 6.6% of patients in the DSP (4 mg) group had low cortisol levels, compared to 1.3% of patients in the placebo group. At week 28, the incidence was similar in the placebo and DSP (4 mg) groups, but lower in the TLC599 group (1.7%).

[0129] Overall, no clinically significant serum cortisol levels exceeding the normal range were observed throughout the study. The results indicate no cumulative effect on the HPA axis following repeated TLC599 injections, and no patients with low cortisol levels reported any treatment-emergent adverse events (TEAEs) related to adrenal insufficiency throughout the study period. Specifically, no events were recorded using the following preferred terms: adrenal insufficiency, adrenal androgen deficiency, adrenal suppression, acute adrenocortical insufficiency, glucocorticoid deficiency, Cushing's syndrome, Cushing's syndrome, hyperadrenalism, or hyperadrenocorticism.

[0130] Table 6. Incidence of low cortisol levels at different time periods – patients who received at least one injection

[0131] a One patient was not included in this count because a morning serum cortisol sample was taken on day 151 (outside the range of this table).

[0132] Table 7. Incidence of low cortisol levels at different time periods - patients receiving a second injection

[0133] In summary, liposomal DSP (TLC599) demonstrated a greater benefit than placebo in reducing ADP and WOMAC pain in patients with knee OA after a single injection lasting 24 weeks or longer. A second injection at 24 weeks extended this benefit to week 52. Furthermore, the safety profile reaffirmed a low incidence of adverse events (AEs), no treatment-related SAEs, and no symptomatic adrenal insufficiency during the two injection periods. These results suggest that TLC599 can provide prolonged efficacy in knee OA without adverse side effects, and that multiple administrations of TLC599 establish a good risk-benefit balance. This disclosure demonstrates the safety and benefit of repeated injections of liposomal DSP in knee OA, and provides an alternative treatment that may benefit OA patients in managing knee OA pain over extended periods, such as 52 weeks or longer.

Claims

1. A method for treating joint pain in a human individual with chronic osteoarthritis, comprising administering repeated doses of a liposomal composition to the human individual via intra-articular delivery, each dose comprising an effective amount of an intra-articular corticosteroid or a pharmaceutically acceptable salt thereof, and a lipid mixture, wherein the repeated doses of the liposomal composition do not induce signs and / or symptoms of adrenal insufficiency in the human individual.

2. The method of claim 1, wherein repeated doses of the liposome composition are administered at dosing intervals selected from the group consisting of: 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, and 36 weeks.

3. The method of claim 1, wherein the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is released during a period selected from the group consisting of: 52 weeks, 53 weeks, 54 weeks, 55 weeks, 56 weeks, 57 weeks, 58 weeks, 59 weeks, 60 weeks, 61 weeks, 62 weeks, 63 weeks, 64 weeks, 65 weeks, 66 weeks, 67 weeks, 68 weeks, 69 weeks, 70 weeks, 71 weeks, 72 weeks, 73 weeks, 74 weeks, 75 weeks, 76 weeks, 77 weeks, 78 weeks, 79 weeks, and 80 weeks.

4. The method of claim 1, wherein the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is released at a rate that does not adversely inhibit the hypothalamic-pituitary-adrenal (HPA) axis.

5. The method of claim 1, wherein the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is released at a rate that does not adversely inhibit the cortisol levels of the human individual.

6. The method of claim 1, wherein the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is selected from the group consisting of: dexamethasone sodium phosphate, dexamethasone, betamethasone, betamethasone sodium phosphate, betamethasone acetate, betamethasone dipropionate, betamethasone valerate, mometasone furoate, triamcinolone, hexamethasone diacetate, triamcinolone diacetate, methylprednisolone sodium succinate, methylprednisolone acetate, hydrocortisone butyrate, hydrocortisone acetate, aclomethasone dipropionate, halcinonide, fluocinolone, fluocinolone acetonide, and combinations thereof.

7. The method of claim 1, wherein the intra-articular corticosteroid is dexamethasone sodium phosphate (DSP) and has an effective amount of about 6 mg to 18 mg.

8. The method of claim 7, wherein the effective amount of DSP is about 12 mg.

9. The method of claim 1, wherein the lipid mixture comprises dioleoylphosphatidylcholine (DOPC) and dioleoylphosphatidylglycerol (DOPG).

10. The method of claim 9, wherein the lipid mixture further comprises about 10 to 33 molar percentages of cholesterol based on the total amount of the lipid mixture.

11. The method of claim 1, wherein the osteoarthritis is knee osteoarthritis.

12. A dosing regimen for the treatment of long-term joint pain, comprising: repeated doses of a liposomal composition administered intra-articularly to the joints of a patient with osteoarthritis, each dose comprising an effective amount of an intra-articular corticosteroid and a lipid mixture, wherein the dosing regimen does not induce signs and / or symptoms of adrenal insufficiency in the patient with osteoarthritis.

13. The dosing regimen of claim 12, wherein repeated doses of the liposome composition are administered at dosing intervals selected from the group consisting of: 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, 24 weeks, 25 weeks, 26 weeks, 27 weeks, 28 weeks, 29 weeks, 30 weeks, 31 weeks, 32 weeks, 33 weeks, 34 weeks, 35 weeks, and 36 weeks.

14. The dosing regimen of claim 12, wherein the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is released during a period selected from the group consisting of: weeks 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, and 80.

15. The dosing regimen of claim 12, wherein the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is released at a rate that does not adversely inhibit the hypothalamic-pituitary-adrenal (HPA) axis.

16. The dosing regimen of claim 12, wherein the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is released at a rate that does not adversely inhibit the cortisol levels of the human individual.

17. The dosing regimen of claim 12, wherein the intra-articular corticosteroid or a pharmaceutically acceptable salt thereof is selected from the group consisting of: dexamethasone sodium phosphate, dexamethasone, betamethasone, betamethasone sodium phosphate, betamethasone acetate, betamethasone dipropionate, betamethasone valerate, mometasone furoate, triamcinolone, hexamethasone diacetate, triamcinolone diacetate, methylprednisolone sodium succinate, methylprednisolone acetate, hydrocortisone butyrate, hydrocortisone acetate, aclomethasone dipropionate, halcinonide, fluocinolone, fluocinolone acetonide, and combinations thereof.

18. The dosing regimen of claim 12, wherein the intra-articular corticosteroid is dexamethasone sodium phosphate (DSP) and has an effective amount of about 6 mg to 18 mg.

19. The dosing regimen of claim 18, wherein the effective amount of DSP is about 12 mg.

20. The dosing regimen of claim 12, wherein the lipid mixture comprises dioleoylphosphatidylcholine (DOPC) and dioleoylphosphatidylglycerol (DOPG).

21. The dosing regimen of claim 20, wherein the lipid mixture further comprises about 10 to 33 molar percentages of cholesterol, based on the total amount of the lipid mixture.

22. The dosing regimen of claim 12, wherein the osteoarthritis patient is a patient with knee osteoarthritis.

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