Bottle-brush polymer for the management of joint pain

Abstract

Pain is a common subjective symptom in subjects with osteoarthritis (OA). Pain typically is treated with acetaminophen or non-steroidal anti-inflammatory drugs (NSAIDs), and two types of intraarticular treatments, corticosteroids and hyaluronic acid (HA) products, also are used for pain control. However, corticosteroid injections have been associated with further cartilage degeneration. and multiple clinical trials failed to demonstrate clinically meaningful efficacy of HA products. There is thus a need for novel approach for the treatment of joint paint. The present disclosure relates to a method for reducing or alleviating joint damage, injury, and / or pain in a subject in need thereof, comprising administering an effective amount of a composition containing a bottle-brush (BB) polymer and that is free of HA. The composition was shown to be associated with functional benefits on pain and the nociceptive process, improving the central nervous system's plasticity and the immune microenvironment in the joint.

Description

[0001] BOTTLE-BRUSH POLYMER FOR THE MANAGEMENT OF JOINT PAIN CROSS REFERENCE TO RELATED APPLICATIONS

[0002] The present application claims the benefit of U. S. provisional patent application serial No.

[0003] 63 / 733,030, filed on December 12, 2024, which is incorporated herein by reference in its entirety.

[0004] TECHNICAL FIELD

[0005] The present invention generally relates to the field of joint disease, and more particularly to the management of joint damages, alterations and associated pain.

[0006] BACKGROUND ART

[0007] Joint pain can be caused by a variety of reasons, including sometimes non-pathologic reasons, but the most common cause is arthritis. According to the Center for Disease Control and Prevention March 2017 Vital Signs announced data estimating that 54.4 million U. S. adults suffer from arthritis, equating to about 25% of the population. The most common types of arthritis are osteoarthritis (OA), rheumatoid arthritis (RA), and gout. The data by the Center for Disease Control and Prevention also indicate that there are approximately 27 million OA and 1.3 million RA patients in the U. S. alone. Joint injury is another major cause of joint pain.

[0008] Osteoarthritis is the most common disease of the joints, and one of the most widespread of all chronic diseases. In the US, OA is second only to heart disease as a cause of work disability in men over 50 years of age. Osteoarthritis was the 6th leading cause of years living with disability at a global level, accounting for 3% of the total global years of living with disability (Woolf 2003).

[0009] Approximately 80% of OA subjects need treatment in the knee. Within the knee joint of symptomatic individuals, the most common radiographic pattern for OA is loss of articular (hyaline) cartilage, deeper cartilaginous remodeling and underlying subchondral bone remodeling, often with osteophyte formation at the joint margins. Both the tibio-femoral and patello-femoral compartments are commonly affected by degenerative changes. Patello-femoral arthritis due to loss of cartilage of the patella and trochlear groove is common in subjects with knee OA, whether or not other joint compartments have OA (Davies 2002, Lankhorst 2017).

[0010] Pain is a common subjective symptom in subjects with OA. Pain typically is treated with acetaminophen or non-steroidal anti-inflammatory drugs (NSAIDs). In addition, two types of intraarticular treatments, corticosteroids and hyaluronic acid (HA) products, also are used for pain control.

[0011] Corticosteroid injections have been associated with further cartilage degeneration (McAlindon 2017), and are used less commonly than they used to be.

[0012] The HA products, or "viscosupplementation" products, are injected into the knee either in once-weekly divided doses (2 cc) for 3 weeks, or in a single larger dose (6 cc; Chevalier 2010). It is claimed for joint pain reduction for weeks to months (Cohen 1998), but multiple clinical trials failed to demonstrate their clinically meaningful efficacy, and thus such treatments are not recommended by the American Academy of Orthopedic Surgeons (AAOS). Indeed, injection with highly viscous supplements (that is, viscosupplementation) offers one approach to re-establishing the rheological and tribological properties of synovial fluid in OA. Badly, this approach has disappointing clinical outcome, either with HA injection alone or enhanced with additional materials (such as glucocorticoids and antioxidants), owing to limited intra-articular residence time and ineffective mechanisms of chondroprotection.

[0013] There is thus a need for novel approach for the treatment of patients with joint paint, such as patients suffering from OA.

[0014] The present description refers to a number of documents, the content of which is herein incorporated by reference in their entirety.

[0015] SUMMARY

[0016] In various aspects and embodiments, the present disclosure provides the following embodiments:

[0017] 1. A method for preventing or reducing joint damage and / or pain in a subject suffering from a joint disease or condition, the method comprising administering to the subject an effective amount of a composition comprising a bottle-brush (BB) polymer comprising a polymeric backbone made of (A) repeat units bearing a polymeric pendant chain and (B) repeat unit free of the polymeric pendant chain, wherein the composition is free of hyaluronic acid (HA) and is not co-administered with a composition comprising HA.

[0018] 2. The method according to embodiment 1, wherein the backbone of the BB polymer is acrylate or methacrylate based.

[0019] 3. The method according to embodiment 1 or 2, wherein the backbone of the BB polymer is poly(methacrylate) ora poly(alkyl methacrylate).

[0020] 4. The method according to any one of embodiments 1 to 3, wherein the backbone of the BB polymer is poly(methyl methacrylate).

[0021] 5. The method according to any one of embodiments 1 to 4, wherein the polymeric pendant chain is attached directly to the backbone of the BB polymer or attached via a linking group.

[0022] 6. The method according to embodiment 5, wherein the polymeric pendant chain is attached to the backbone of the BB polymer via the linking group.

[0023] 7. The method according to embodiment 5 or 6, wherein the linking group attaching the polymeric pendant chain to the backbone of the BB polymer is a carboxylic acid, an ester, an amine, an azide, or a thiol functional group, or an alkylene, alkenylene, or alkylene group that is interrupted or not with one or more ester, amine, or azide, or thiol functional group, preferably the linking group is *-CH2-CH2-O-C(=O)-#, wherein * is the point of attachment of the backbone and#is the point of attachment to the polymeric pendant chain.

[0024] 8. The method according to any one of embodiments 1 to 7, wherein the polymeric pendant chain is zwitterionic.

[0025] 9. The method according to any one of embodiments 1 to 8, wherein the polymeric pendant chains comprise repeat units (D) having optionally attached thereto a substituent, either directly or through a linking group.

[0026] 10. The method according to embodiment 9, wherein repeat units (D) are repeat units of:

[0027] • poly(acrylic acid), such as a poly(alkyl acrylic acid),

[0028] • a poly(acrylate), such as a poly(alkyl acrylate), for example poly(methyl acrylate), • poly(methacrylic acid), such as poly(alkyl methacrylic acid), for example poly(methyl methacrylic acid),

[0029] • a poly(methacrylate), such as a poly(alkyl methacrylate), for example poly(methyl methacrylate),

[0030] • poly(acrylamide), or

[0031] • a poly(methacrylamide), such as a poly(alkyl methacrylate), for example poly(methyl methacrylate)

[0032] preferably repeat units of a poly(methacrylate).

[0033] 11. The method according to embodiment 9 or 10, wherein repeat units (D) have attached thereto a substituent either directly or through the linking group.

[0034] 12. The method according to any one of embodiments 9 to 11, wherein the substituent is attached to repeat units (D) through the linking group.

[0035] 13. The method according to anyone of embodiments 9 to 12, wherein the linking group attaching the substituent to the repeat units (D) is a carboxylic acid, an ester, an amine, an azide, or a thiol functional group, or an alkylene, alkenylene, or alkylene group that is interrupted or not with one or more ester, amine, or azide, or thiol functional group, preferably the linking group is -CH2-CH2-.

[0036] 14. The method according to any one of embodiments 9 to 13, wherein the substituent attached to repeat units (D) is a phosphorylcholine group, a saccharide or disaccharide group including but not limited to glucose, sucrose, lactose and their derivatives, such as D-gluconolactone and lactobionolactone, or a biocompatible hydrophilic group such as hydroxy, oligo(ethylene oxide), carboxy, amino, sulfo, thiol, phosphate, or a derivative thereof. The method according to any one of embodiments 9 to 14, wherein the substituent attached to repeat units (D) is phosphorylcholine.

[0037] The method according to any one of embodiments 1 to 15, wherein the polymeric pendant chain is poly(2-methacryloyloxyethyl phosphorylcholine) of formula:

[0038]

[0039] The method according to any one of embodiments 1 to 16, wherein the BB polymer comprises repeat units of formula:

[0040]

[0041] wherein R is H, an alkyl, a halogen (e.g., Br, Cl), or a peptide (e.g., a peptide comprising one or more cysteine residues), and

[0042] wherein x represents the number of repeat unit and preferably varies between 1 and 1000, more preferably between 10 and 500, yet more preferably between 20 and 250, and most preferably between 25 and 100.

[0043] 18. The method according to any one of embodiments 1 to 17, wherein the BB polymer further comprises (C) repeat units bearing another pendant group. 19. The method according to embodiment 18, wherein said other pendant group in repeat units (C) is an alkylene, alkenylene, or alkynylene (preferably alkylene), each of which being optionally interrupted or terminated by one or more of OH, ammonium accompanied by a pharmaceutically acceptable counterion, thiol, amine, carboxyl, and / or azide, preferably OH or said ammonium.

[0044] 20. The method according to embodiment 19, wherein the alkylene comprises between 1 and 20 carbon atoms, preferably between 1 and 8 carbon atoms, and most preferably 2 carbon atoms and wherein the alkenylene and alkynylene comprise between 2 and 20 carbon atoms, preferably between 2 and 8 carbon atoms, and most preferably 2 carbon atoms.

[0045] 21. The method according to embodiment 19 or 20, wherein said other pendant groups in repeat units (C) is an alkylene, alkenylene, and alkynylene group (preferably alkylene), each of which being terminated by OH, ammonium accompanied by a pharmaceutically acceptable counterion, thiol, amine, carboxyl, and / or azide, preferably OH or said ammonium.

[0046] 22. The method according to any one of embodiments 19 to 21, wherein said ammonium group is -N+(CH3)2(alkyl), wherein the alkyl comprises between 1 and 20 carbon atoms, preferably between 1 and 8 carbon atoms, and most preferably 2 carbon atoms; preferably the ammonium group is preferably accompanied by a halogenide ion, preferably bromide (Br).

[0047] 23. The method according to any one of embodiments 19 to 22, wherein said other pendant groups in repeat units (C) is -CH2-CH2-OH and -CH2-CH2-N+(CH3)2(CH2CH3) Br-.

[0048] 24. The method according to any one of embodiments 19 to 23, wherein the BB polymer is a triblock copolymer comprising a first block, a second block and a third block, the second block being located between the first block and the third block, the first block and the third block comprising statistically arranged repeat units B and C, while the second block comprises statistically arranged repeat units A and B.

[0049] 25. The method according to any one of embodiments 1 to 24, wherein the backbone of the BB polymer further comprises a remnant of an initiator molecule.

[0050] 26. The method according to any one of embodiments 1 to 25, wherein the BB polymer further comprises one or more capping blocks.

[0051] 27. The method according to any one of embodiments 1 to 26, wherein the BB polymer comprises:

[0052] - repeat units (A) that are: o ^-o

[0053] , wherein x and R are as defined above,

[0054]

[0055] o

[0056] - optionally repeat units (C) that are

[0057]

[0058] OH

[0059] The method according to any one of embodiments 1 to 27, wherein the BB polymer comprises the following repeat units: (A), preferably

[0060]

[0061] and (B)

[0062] wherein x and R are as defined above;

[0063] preferably with a grafting ratio between about 40% and about 60%, more preferably between about 45% and about 55%, and most of about 45% or about 55%.

[0064] The method according to any one of embodiments 1 to 27, wherein the BB polymer comprises the following repeat units (A) preferably O^'-'O, wherein x and R are as defined above,

[0065]

[0066] preferably the BB polymer is of formula:

[0067]

[0068] preferably:

[0069]

[0070] preferably:

[0071] wherein x is as defined above and m and n represent repeat unit ratios (e.g., m = number of repeat units of the formula on the left

[0072] and, 0 < m < 1 and 0 ≤ n <1;

[0073] total number of repeat units in polymer

[0074] preferably having a grafting ratio between about 30% and about 100%; more preferably the grafting ratio is about 30%, about 35%, about 40%, about 45%, or about 50% or more and / or about 100%, about 90%, about 80%, about 70%, about 65%, about 60%, or about 55% or about 50% less.

[0075] 29. The method according to any one of embodiments 1 to 28, wherein the BB polymer is (PBiBEM54o-g-PMPC28)-stat-PHEMA6o-stat-PMMAeoo or (PBiBEM456-g-PMPC35)-stat- PHEMA3-stat-PMMA37o.

[0076] 30. The method of embodiment 24, wherein the triblock copolymer comprises repeat units:

[0077] (A) preferably

[0078] , wherein x and R are as defined above,

[0079] (

[0080]

[0081] B) preferably the BB polymer is of formula:

[0082]

[0083] wherein x and R are as defined above, and wherein a, b, c, d, e, and f are ≥ 1 and represent the number of repeat units,

[0084] preferably:

[0085] a varies between about 25 and about 500, preferably between about 50 and about 200, more preferably between 50 and 100,

[0086] b varies between about 25 and about 500, preferably between about 50 and about 200, more preferably between 100 and 150, c varies between about 100 and about 1000, preferably between about 200 and about 800, preferably between about 400 and about 500,

[0087] d varies between about 100 and about 450, preferably between about 200 and about 800, preferably between about 350 and about 500,

[0088] e varies between about 25 and about 500, preferably between about 50 and about 200, more preferably between 100 and 150, and

[0089] f varies about 25 and about 500, preferably between about 50 and about 200, more preferably between 50 and 100;

[0090] more preferably the BB polymer has a grafting ratio between about 25% and about 75%, and most preferably the BB polymer is of formula:

[0091]

[0092] 31. The method according to any one of embodiments 1 to 30, wherein the molecular weight of the backbone is about 10 kDa, about 20 kDa, about 30 kDa, about 40 kDa, about 50 kDa, about 60 kDa, about 70 kDa, about 80 kDa, about 100 kDa, about 200 kDa, about 300 kDa, about 400 kDa, about 500 kDa, about 750 kDa, or about 900 kDa or more and / or about 1000 kDa, about 750 kDa, about 500 kDa, about 400 kDa, about 300 kDa, about 200 kDa, or about 100 kDa or less.

[0093] 32. The method according to any one of embodiments 1 to 31, wherein the molecular weight of the backbone of the BB polymer is about 90 kDa or 100 kDa.

[0094] 33. The method according to any one of embodiments 1 to 32, wherein the molecular weight of the polymeric pendant chain is about 1 kDa, about 5 kDa, about 10 kDa, about 20 kDa, about 30 kDa, about 40 kDa, about 50 kDa, about 60 kDa, about 70 kDa, about 80 kDa, or about 90 kDa or more and / or about 100 kDa, about 90 kDa, about 80 kDa, about 70 kDa, about 60 kDa, about 50 kDa, about 40 kDa, about 30 kDa, about 20 kDa, or about 15 kDa or less. 34. The method according to any one of embodiments 1 to 33, wherein the molecular weight of the polymeric pendant chain is about 15 kDa or is between about 10 kDa to about 20 kDa.

[0095] 35. The method according to any one of embodiments 1 to 34, wherein a grafting ratio of the BB polymer is between about 30 and about 100 %.

[0096] 36. The method according to any one of embodiments 1 to 35, wherein a grafting ratio of the BB polymer is about 30%, about 35%, about 40%, about 45%, or about 50% or more and / or about 100%, about 90%, about 80%, about 70%, about 65%, about 60%, about 55%, or about 50% or less.

[0097] 37. The method according to any one of embodiments 1 to 36, wherein the grafting ratio of the BB polymer is between about 40 and about 60%, preferably between about 45% and about 55%, most preferably is about 45% or about 55%.

[0098] 38. The method according to any one of embodiments 1 to 37, wherein the concentration of the BB polymer in the composition is about 1, about 25, about 50, about 75, about 85, about 90, about 95, about 100, about 150, about 200, about 250, about 300, about 350, about 400, about 450, or about 500 pg / ml or more and / or about 40, 35, 30, 25, 20, 15, 10, about 5, about 1, about 0.5, about 0.25, or about 0.1 mg / mL or less.

[0099] 39. The method according to any one of embodiments 1 to 38, wherein the concentration of the BB polymer is about 20 mg / ml to about 40 mg / ml.

[0100] 40. The method according to any one of embodiments 1 to 39, wherein the composition is free of linear polymer.

[0101] 41. The method according to any one of embodiments 1 to 40, wherein the composition further comprises a linear polymer.

[0102] 42. The method according to embodiment 41, wherein the linear polymer is dextran, poly(vinylpyrrolidone), polyethylene glycol), hydroxypropyl cellulose, a polymethacrylate polymer or copolymer, a polyoxazoline polymer, or a polyacrylate polymer or copolymer, or a (preferably pharmaceutically acceptable) salt thereof.

[0103] 43. The method according to embodiment 42, wherein the linear polymer is poly(vinylpyrrolidone), or a (preferably pharmaceutically acceptable) salt thereof.

[0104] 44. The method according any one of embodiments 41 to 43, wherein the linear polymer is partially crosslinked.

[0105] 45. The method according to any one of embodiments 41 to 44, wherein the linear polymer has a molecular weight of about 5 kDa, about 10 kDa, about 25 kDa, about 50 kDa, about 100 kDa, about 250 kDa, or about 500 kDa or more and / or 10 MDa, about 8 MDa, or about 5 MDa or less.

[0106] 46. The method according to any one of embodiments 41 to 45, comprising the BB polymer and the linear polymer in a BB polymer: linear polymer weight ratio between about 1:1 and about 1:20, preferably between about 1:2.5 and about 1:15, more preferably between about 1:5 and about 1:10.

[0107] 47. The method according to any one of embodiments 1 to 46, wherein the composition further comprises a pharmaceutically acceptable carrier or excipient.

[0108] 48. The method according to any one of embodiments 1 to 46, wherein the pharmaceutically acceptable carrier or excipient comprises a saline solution, preferably isotonic, preferably buffered at a pH of about 7 to about 7.4.

[0109] 49. The method according to any one of embodiments 1 to 46, wherein the pharmaceutically acceptable carrier or excipient comprises phosphate-buffered saline (PBS).

[0110] 50. The method according to any one of embodiments 47 to 49, wherein the pharmaceutically acceptable carrier or excipient comprises one or more additives such as preservatives, stabilizers, antioxidants, surfactants, viscosity modifiers, and / or chelating agents.

[0111] 51. The method according to any one of embodiments 1 to 50, wherein the composition further comprises one or more additional therapeutic agents, or wherein the method further comprises administering one or more additional therapeutic agents.

[0112] 52. The method according to embodiment 51, wherein the one or more additional therapeutic agents comprise an analgesic, an anti-inflammatory agent, an antioxidant or a biologic. 53. The method according to embodiment 52, wherein the anti-inflammatory agent comprises a nonsteroidal anti-inflammatory drug (NSAID) ora glucocorticoid.

[0113] 54. The method according to any one of embodiments 1 to 53, wherein the joint disease or condition is joint injury or arthritis.

[0114] 55. The method according to embodiment 54, wherein the joint disease or condition is osteoarthritis (OA).

[0115] 56. The method according to any one of embodiments 1 to 55, wherein the joint damage and / or pain is knee joint damage and / or pain.

[0116] 57. The method according to any one of embodiments 1 to 56, wherein the composition is injected in the joint or in the vicinity of the joint.

[0117] 58. The method according to any one of embodiments 1 to 57, wherein the subject is a human. 59. The method according to any one of embodiments 1 to 58, wherein the subject is a companion animal, such as a dog or a cat, or a farm animal such as an ovine, caprine or bovine, or a sport animal such as a horse.

[0118] 60. A composition as defined in any one of embodiments 1 to 54, for use in preventing or reducing joint damage and / or pain in a subject suffering from a joint disease or condition.

[0119] 61. The composition for use according to embodiment 60, wherein the joint disease or condition is joint injury or arthritis.

[0120] 62. The composition for use according to embodiment 61, wherein the joint disease or condition is osteoarthritis.

[0121] 63. The composition for use according to any one of embodiments 60 to 62, wherein the joint damage and / or pain is knee joint damage and / or pain.

[0122] 64. The composition for use according to any one of embodiments 60 to 63, wherein the composition is for injection in the joint or in the vicinity of the joint.

[0123] 65. The composition for use according to any one of embodiments 60 to 64, wherein the subject is a human.

[0124] 66. The composition for use according to any one of embodiments 60 to 64, wherein the subject is a companion animal, such as a dog or a cat, or a farm animal such as an ovine, caprine or bovine, or a sport animal such as a horse.

[0125] 67. Use of a composition as defined in any one of embodiments 1 to 53, for preventing or reducing joint damage and / or pain in a subject suffering from a joint disease or condition.

[0126] 68. Use of a composition as defined in any one of embodiments 1 to 53, for the manufacture of a medicament for preventing or reducing joint damage and / or pain in a subject suffering from a joint disease or condition.

[0127] 69. The use according to embodiment 67 or 68, wherein the joint disease or condition is joint injury or arthritis.

[0128] 70. The use according to any one of embodiments 67 to 69, wherein the joint disease or condition is osteoarthritis.

[0129] 71. The use according to any one of embodiments 67 to 70, wherein the joint damage and / or pain is knee joint damage and / or pain.

[0130] 72. The use according to any one of embodiments 67 to 71, wherein the composition is for injection in the joint or in the vicinity of the joint.

[0131] 73. The use according to any one of embodiments 67 to 72, wherein the subject is a human. 74. The use according to any one of embodiments 67 to 72, wherein the subject is a companion animal, such as a dog or a cat, or a farm animal such as an ovine, caprine or bovine, or a sport animal such as a horse.

[0132] Other objects, advantages and features of the present disclosure will become more apparent upon reading of the following non-restrictive description of specific embodiments thereof, given by way of example only with reference to the accompanying drawings.

[0133] BRIEF DESCRIPTION OF DRAWINGS

[0134] In the appended drawings:

[0135] FIG. 1 depicts the Total histological OsteoArthritis Research Society International (OARSI) score at day 42 and day 84 in the different treatment groups.

[0136] FIG. 2 depicts the Cartilage degeneration score at day 42 and day 84 in the different treatment groups.

[0137] FIG. 3 depicts the Osteophyte score at day 42 and day 84 in the different treatment groups.

[0138] FIG. 4 depicts the Calcified cartilage and subchondral bone damage score at day 42 and day 84 in the different treatment groups.

[0139] FIG. 5 depicts the Synovitis score at day 42 and day 84 in the different treatment groups.

[0140] FIG. 6 depicts the Loss of proteoglycans score at day 42 and day 84 in the different treatment groups.

[0141] FIGs. 7A and 7B show the percentage of total altered surface and severity score at day 42 and day 84 in the different treatment groups.

[0142] FIG. 8 shows pictures of a stifle of a naive rat (upper pictures) and MI-RAT placebo rat (lower pictures) at day 84.

[0143] FIG. 9 shows the spontaneous expression of pain overtime as assessed by the distribution of static weight-bearing (SWB) on the hind limbs in the different treatment groups. An incapacitance Meter® (IITC Life Science) was used to measure the SWB on the hind limbs, readings are taken over a period of 3 seconds and expressed in grams (Gervais., et al. 2019, Keita-Alassane., et al. 2022). A total of three readings was taken for each limb of the rat, at each assessment time, for this spontaneous expression of pain. Statistical analysis: mixed model on the SWB asymmetry index. * Naive different of all 4 MI-RAT groups at this timepoint; ** Naive different of MI-RAT HA, MI-RAT BB-HA and MI-RAT Placebo at this timepoint. Different letters indicate significant intra- or inter-group differences.

[0144] FIG. 10 shows the evoked expression of pain over time as assessed by the paw withdrawal threshold (PWT) following tactile mechanical stimulation on both plantar paws in the different treatment groups. An electronic von Frey® esthesiometer (IITC Life Science) was used to measure the PWT following tactile mechanical stimulation on both plantar paws. For each animal, 3 measurements of each hind paw were carried out with an interval of 60 s between stimuli. Actions such as vocalization, agitation, jumping, and avoidance were considered indicative of a withdrawal reaction. The peak force in grams, to this evoked response, was recorded (Gervais., et al. 2019, Keita-Alassane., et al. 2022). Statistical analysis: mixed model on the PWT asymmetry index. * Naive different of all 4 MI-RAT groups at this timepoint. Different letters indicate significant intra- or inter-group differences.

[0145] FIG. 11 shows the pain central sensitization over time in the different treatment groups. Response to mechanical temporal summation (RMTS) (TopCat Metrology Ltd.) was used for measuring the number of repetitive mechanical stimulations generating avoidance or aversive behavior, with intensity of 4N, stimulation frequency of 0.4 Hz, period between each stimulation of 2.5 s and duration of each stimulation of 1.5 s (cut-off number of stimulations n=30) (Guillot 2014 and Monteiro 2016). Statistical analysis: mixed model on count number of stimulation. * Naive different of all 4 MI-RAT groups at this timepoint.

[0146] FIGs. 12A-B show the pain endogenous inhibition over time in the different treatment groups. Endogenous pain control was evaluated with electronic von Frey® esthesiometer (I ITC Life Science) to measure the paw withdrawal threshold (PWT) with conditioning stimulus, ear clip for 1 minute, following tactile mechanical stimulation on both hind paws (PWT post-CPM). For each animal, 3 measurements of each hind paw were carried out with an interval of 60 s between stimuli. Actions such as vocalization, agitation, jumping, and avoidance were considered indicative of the withdrawal reaction. The peak force in grams was recorded. Statistical analysis: mixed model on the functionality percentage between PWT pre- and post-CPM for each hind paw, right hind paw (RHP) and left hind paw (LHP). RHP (FIG. 12A) is in direct link with the primary source of pain (OA induction in the right stifle), while LHP sensitivity (FIG. 12B) is tested for determining a possible central nervous system participation. FIG. 12A: * Naive different of MI-RAT Placebo and MI-RAT HA at this timepoint. ** Naive different of MI-RAT Placebo at this timepoint. Different letters indicate significant intra-group differences; FIG. 12B: * MI-RAT Placebo different of MI-RAT BB & BB-HA at this timepoint. Different letters indicate significant intra-group differences.

[0147] FIGs. 13A-B show the pain facilitation overtime in the different treatment groups. Central sensitization was evaluated with electronic von Frey® esthesiometer (IITC Life Science) to measure the paw withdrawal threshold (PWT) after RMTS, following tactile mechanical stimulation on both plantar paws (PWT post-RMTS). For each animal, 3 measurements of each hind paw were carried out with an interval of 60 s between stimuli. Actions such as vocalization, agitation, jumping, and avoidance were considered indicative of the withdrawal reaction. The peak force in grams was recorded. Statistical analysis: mixed model on the functionality percentage between PWT pre- and post-RMTS for each hind paw, right hind paw (RHP) and left hind paw (LHP). RHP (FIG. 13A) is in direct link with the primary source of pain (OA induction in the right stifle) while LHP sensitivity (FIG. 13B) is tested for determining a possible central nervous system participation. FIG. 13A: * MI-RAT Placebo different of MI-RAT HA and MI-RAT BB-HA at this timepoint; ** Naive different of MI-RAT Placebo and MI-RAT HA at this timepoint. Different letters indicate significant intra-group differences for MI-RAT HA and BB-HA. FIG 13B: * Naive different of MI-RAT HA at this timepoint; ** MI-RAT HA different of MI-RAT BB and BB-HA and at this timepoint. Different letters indicate significant intra-group differences for MI-RAT HA and BB-HA.

[0148] FIGs. 14A-B show the results of a principal component analysis (PCA) of the neuropeptidome at day 42 (FIG. 14A) and day 84 (FIG. 14B) in the different treatment groups.

[0149] FIG. 15 shows the architecture of a bottle-brush polymer according to the present disclosure.

[0150] DETAILED DISCLOSURE

[0151] The use of the terms "a" and "an" and "the" and similar referents in the context of describing the technology (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0152] The terms "comprising", "having", "including", and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to") unless otherwise noted.

[0153] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context.

[0154] The use of any and all examples, or exemplary language (“e.g.”, "such as") provided herein, is intended merely to better illustrate embodiments of the claimed technology and does not pose a limitation on the scope unless otherwise claimed.

[0155] No language in the specification should be construed as indicating any non-claimed element as essential to the practice of embodiments of the claimed technology.

[0156] Herein, the term "about" has its ordinary meaning. The term “about” is used to indicate that a value includes an inherent variation of error for the device or the method being employed to determine the value, or encompass values close to the recited values, for example within 10% of the recited values (or range of values).

[0157] Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All subsets of values within the ranges are also incorporated into the specification as if they were individually recited herein.

[0158] Where features or aspects of the disclosure are described in terms of Markush groups or list of alternatives, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member, or subgroup of members, of the Markush group or list of alternatives. Unless specifically defined otherwise, all technical and scientific terms used herein shall be taken to have the same meaning as commonly understood by one of ordinary skill in the art (e.g., in stem cell biology, cell culture, molecular genetics, immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0159] Unless otherwise indicated, the recombinant protein, cell culture, and immunological techniques utilized in the present disclosure are standard procedures, well known to those skilled in the art. Such techniques are described and explained throughout the literature in sources such as, J. Perbal, A Practical Guide to Molecular Cloning, John Wiley and Sons (1984), J. Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbour Laboratory Press (1989), T. A. Brown (editor), Essential Molecular Biology: A Practical Approach, Volumes 1 and 2, IRL Press (1991), D. M. Glover and B. D. Hames (editors), DNA Cloning: A Practical Approach, Volumes 1-4, IRL Press (1995 and 1996), and F. M. Ausubel et al. (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-lnterscience (1988, including all updates until present), Ed Harlow and David Lane (editors) Antibodies: A Laboratory Manual, Cold Spring Harbour Laboratory, (1988), and J. E. Coligan et al. (editors) Current Protocols in Immunology, John Wiley & Sons (including all updates until present).

[0160] As used herein, the term “subject” can be a vertebrate, such as a mammal, a fish, a bird, a reptile, or an amphibian. Thus, the subject can be a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig or rodent. The term does not denote a particular age or sex. Thus, adult and newborn subjects, as well as fetuses, whether male or female, are intended to be covered. In one aspect, the subject is a mammal. A patient refers to a subject afflicted with an ailment, disease, or disorder. The term “patient” includes human and veterinary subjects.

[0161] Tribosupplementation is the delivery other than an exogenous HA-based lubricant to a synovial joint, with the intention of improving cartilage lubrication as a treatment for OA. The present inventors are proposing that their biomimetic boundary lubricant, a bottle-brush (BB) polymer, to bind and lubricate articular cartilage. In the studies described herein, the present inventors have demonstrated in a rat model of OA that the administration of a BB polymer solution protects the stifle articular structures, which is translated in functional benefits on both direct pain (either spontaneous or evoked) and on nociceptive process of facilitation / inhibitory endogenous control of nociception. The BB polymer solution was superior to solutions comprising HA or a HA-BB mixture in several aspects.

[0162] The BB polymer solution according to the present disclosure is associated with functional benefits on both direct pain (either spontaneous or evoked) expression and on nociceptive process of facilitation / inhibitory endogenous control, associated to no solicitation of either facilitation or inhibition process, which could be translated on long-term to a better preservation of the central nervous system plasticity. The BB polymer solution according to the present disclosure may be associated with an improvement of the immune microenvironment in the joint. Inside the joint, macrophages are polarised into M1 pro-inflammatory phenotype and M2 anti-inflammatory phenotype under stimulation of different factors, which release and regulate inflammatory response and cartilage growth. Accumulating studies have tried to alleviate OA by regulating macrophage homeostasis, and based on some preliminary results obtained by the present inventors, the intraarticular injection of BB polymer solution according to the present disclosure leads to such effect of reduced synovial macrophage polarisation, either on the conventionally recognized binary states (M1 / M2) or other functional phenotypes.

[0163] Thus, in an aspect, the present disclosure relates a method for reducing or alleviating joint damage, injury and / or pain in a subject in need thereof, the method comprising administering to the subject an effective amount of a composition comprising a BB polymer comprising a polymeric backbone with polymeric pendant chains, wherein the composition is free of HA and is not coadministered with a composition comprising HA. The present disclosure also relates to the use of a composition comprising a BB polymer comprising a polymeric backbone with polymeric pendant chains, wherein the composition is free of HA and is not for use with a composition comprising HA. The present disclosure also relates to the use of a composition comprising a BB polymer comprising a polymeric backbone with polymeric pendant chains for reducing or alleviating joint damage, injury and / or pain in a subject, wherein the composition is free of HA and is not for use with a composition comprising HA. The present disclosure also relates to the use of a composition comprising a BB polymer comprising a polymeric backbone with polymeric pendant chains for the manufacture of a medicament for reducing or alleviating joint damage, injury and / or pain in a subject, wherein the composition is free of HA and is not for use with a composition comprising HA. The present disclosure also relates to a composition comprising a BB polymer comprising a polymeric backbone with polymeric pendant chains for use in reducing or alleviating joint damage, injury and / or pain in a subject, wherein the composition is free of HA and is not for use with a composition comprising HA.

[0164] The term “bottle-brush (BB) polymer” as used herein refers to a polymer comprising a linear polymeric backbone with multiple polymeric pendant chains attached to the backbone. FIG. 15 shows the typical architecture of a BB polymer. The aforementioned backbone and polymeric pendant chains form the “bottle-brush” block in this polymer. Optional “capping blocks” located at either or both ends of the linear backbone of the “bottle-brush” block are also shown in FIG. 15.

[0165] The BB polymer can be characterized by its grafting ratio. The grafting ratio represents the percentage of repeat units of the backbone of the BB polymer that bear a polymeric pendant chain. Thus, the BB polymer comprises (A) repeat units bearing the polymeric pendant chain and (B) repeat units without the polymeric pendant chain. In other words, the BB polymer is in fact a copolymer. This copolymer can further comprise (C) repeat units bearing other pendant groups. For the BB polymer comprised in the present composition, the grafting ratio typically ranges between about 25 and about 100 %. In preferred embodiments of the disclosure, the grafting ratio is about 30%, about 35%, about 40%, about 45%, or about 50% or more and / or about 100%, about 90%, about 80%, about 70%, about 65%, about 60%, about 55%, or about 50% or less. In most preferred embodiments, the grafting ratio is between about 30 and about 60%, preferably between about 30% and about 55%, preferably between about 32% and about 55%, more preferably is about 35%, 45% or 55%.

[0166] The BB polymer can also be characterized by the molecular weight of its backbone and the molecular weight of its polymeric pendant chains.

[0167] In embodiments, the molecular weight of the backbone is about 10 kDa, about 20 kDa, about 30 kDa, about 40 kDa, about 50 kDa, about 60 kDa, about 70 kDa, about 80 kDa, about 100 kDa, about 200 kDa, about 300 kDa, about 400 kDa, about 500 kDa, about 750 kDa, or about 900 kDa or more and / or about 1000 kDa, about 750 kDa, about 500 kDa, about 400 kDa, about 300 kDa, about 200 kDa, or about 100 kDa or less. In preferred embodiments, the molecular weight of the backbone is about 90 kDa (e.g., 88700 Da). In preferred embodiments, the molecular weight of the backbone is about 100 kDa.

[0168] In embodiments, the molecular weight of the polymeric pendant chain is about 1 kDa, about 5 kDa, about 10 kDa, about 20 kDa, about 30 kDa, about 40 kDa, about 50 kDa, about 60 kDa, about 70 kDa, about 80 kDa, or about 90 kDa or more and / or about 100 kDa, about 90 kDa, about 80 kDa, about 70 kDa, about 60 kDa, about 50 kDa, about 40 kDa, about 30 kDa, about 20 kDa, or about 15 kDa or less. In preferred embodiments, the molecular weight of the polymeric pendant chain is between about 10 kDa to about 20 kDa. In preferred embodiments, the molecular weight of the polymeric pendant chain is about 15 kDa (e.g., 13275 Da).

[0169] The BB polymer is also characterized by the nature of the repeat units in its backbone (i.e. repeat units A, B, and C) and the nature of the polymeric pendant chains.

[0170] The nature of the repeat units of the backbone of the BB polymer is chosen so that it produces polymer (preferably a pharmaceutically acceptable polymer) and allows for post-grafting of the polymeric pendant chains. The grafting of the polymeric pendant chains may be achieved using the “grafted from” or “grafting to” methods. In the “grafted from” approach, the polymeric pendant chains are grown from a macromolecular chain bearing initiator functional groups. In the “grafting to” approach, the pendant chains are polymerized separately and grafted to the main chain afterwards.

[0171] In embodiments, the backbone of the BB polymer may be

[0172] • acrylate based e.g.:

[0173] o poly(acrylic acid), or

[0174] o a poly(acrylate), such as poly(alkyl acrylate), more preferably poly(methyl acrylate), or • methacrylate based e.g.:

[0175] o poly(methacrylic acid), or

[0176] o a poly(methacrylate), such as poly(alkyl methacrylate, more preferably poly(methyl methacrylate).

[0177] In preferred embodiments, the backbone of the BB polymer is poly(methacrylic acid) or a poly(alkyl methacrylate), such as poly(methyl methacrylate). Most preferably, the backbone of the BB polymer is poly(methyl methacrylate).

[0178] As noted above, the BB polymer comprises (A) repeat units bearing the polymeric pendant chain and (B) repeat units without the polymeric pendant chain and can further comprise (C) repeat units bearing other pendant groups. Thus, a BB polymer with a poly(methyl methacrylate) backbone will comprise methyl methacrylate repeat units (A) bearing the polymeric pendant chain, methyl methacrylate repeat units (B) that are not bearing any pendant groups (i.e., they are bare), and optionally methyl methacrylate repeat units (C) being other pendant groups.

[0179] As noted above, the pendant chains of the bottle-brush polymer are polymeric, thus they comprise a polymer. The polymer in the pendant chains may be attached directly to the backbone of the BB polymer or attached via a linking group. Preferably, a linking group is used. Suitable linking groups (for attaching the polymeric pendant chain to the backbone of the BB polymer) include carboxylic acid, ester, amine, azide, and thiol functional groups as well as alkylene, alkenylene, and alkynylene groups, the alkylene, alkenylene, and alkynylene groups being interrupted or not with one or more ester, amine, azide, and / or thiol functional groups. Herein “interrupted” means that a functional group (ester, amine, azide, or thiol) is located at either end or in between two carbon atoms of the alkylene, alkenylene, and alkynylene groups. A preferred linking group is *-CH2-CH2-O-C(=O)-#, wherein * is the point of attachment of the backbone and#is the point of attachment to the polymeric pendant chain.

[0180] In embodiments, the polymeric pendant chains is zwitterionic.

[0181] In embodiments, the polymeric pendant chain comprises repeat units (D) optionally having attached thereto a substituent, directly or through a linking group.

[0182] In embodiments, the repeat units (D) are repeat units of:

[0183] • poly(acrylic acid), such as a poly(alkyl acrylic acid),

[0184] • a poly(acrylate), such as a poly(alkyl acrylate), for example poly(methyl acrylate), • poly(methacrylic acid), such as poly(alkyl methacrylic acid), for example poly(methyl methacrylic acid),

[0185] • a poly(methacrylate), such as a poly(alkyl methacrylate), for example poly(methyl methacrylate),

[0186] • poly(acrylamide), or • a poly(methacrylamide), such as a poly(alkyl methacrylate), for example poly(methyl methacrylate).

[0187] In preferred embodiments, repeat units (D) are repeat units of poly(methacrylate).

[0188] Examples of substituents attached to repeat units (D) include phosphorylcholine ( o'

[0189] O— P i— o +

[0190] ii N: T

[0191]

[0192] o I

[0193] , the open link attached to the oxygen atom on the left indicating the bond attaching the phosphorylcholine to the rest of the molecule), saccharide and disaccharide groups, including but not limited to glucose, sucrose, lactose and their derivatives such as D- gluconolactone and lactobionolactone, as well biocompatible hydrophilic groups such as hydroxy, oligo(ethylene oxide), carboxy, amino, sulfo, thiol, phosphate, and derivatives thereof. Suitable linking groups for attaching the substituent to repeat units (D) include carboxylic acid, ester, amine, azide, and thiol functional groups as well as alkylene, alkenylene, and alkynylene groups, the alkylene, alkenylene, and alkynylene groups being interrupted or not with or more ester, amine, azide, and / or thiol functional groups. A preferred linking group is an alkylene group, preferably -CH2-CH2-.

[0194] In preferred embodiments, repeat units (D) have attached thereto said substituent, preferably through said linking group. In preferred embodiments, the substituent is phosphorylcholine. In preferred embodiments, the linking group is -CH2-CH2-.

[0195] In preferred embodiments, the polymer in the pendant chains is poly(2-methacryloyloxyethyl

[0196] phosphorylcholine of formula: and in embodiments:

[0197]

[0198] , wherein x represents the number of repeat unit (D) and, in embodiments, varies between 1 and 1000. In embodiments, x is preferably varying between 10 and 500, and more preferably between 20 and 250, and most preferably between 25 and 100.

[0199] In most preferred embodiments, the repeat unit (B) comprising the pendant chains is:

[0200]

[0201] is H, an alkyl, a halogen (e.g., Br, Cl), ora peptide (e.g., a peptide comprising one or more cysteine residues),

[0202]

[0203] , wherein x is as defined above.

[0204] As noted above, the BB polymer comprises: (A) repeat units bearing the polymeric pendant chain, (B) repeat units without the polymeric pendant chain, and optionally (C) repeat units bearing other pendant groups. In embodiments, repeat units (C) bearing other pendant groups are present. In alternative embodiments, they are absent. In embodiments, the BB polymer is a triblock copolymer (e.g. the backbone of the BB polymer forms is a triblock copolymer), comprising a first block, a second block and a third block, the second block being located between the first block and the third block, the first block and the third block comprising statistically arranged repeat units B and C, while the second block comprises statistically arranged repeat units A and B. Herein, “statistically arranged” means that the repeat units are arranged in a sequence dictated by the reaction kinetics of the monomers. This term is commonly used interchangeably with “random” in the literature.

[0205] Non-limiting examples of said other pendant groups in repeat units (C) include: alkylene, alkenylene, and alkynylene groups (preferably alkylene), each of which being optionally interrupted or terminated by one or more of OH, ammonium accompanied by a pharmaceutically acceptable counterion, thiol, amine, carboxyl, and / or azide, preferably OH or said ammonium.

[0206] In preferred embodiments, the alkylene comprises between 1 and 20 carbon atoms, preferably between 1 and 8 carbon atoms, and most preferably 2 carbon atoms. In embodiments, the alkenylene and alkynylene comprise between 2 and 20 carbon atoms, preferably between 2 and 8 carbon atoms, and most preferably 2 carbon atoms.

[0207] In preferred embodiments, said other pendant groups in repeat units (C) is alkylene, alkenylene, and alkynylene groups (preferably alkylene), each of which being terminated by OH, ammonium accompanied by a pharmaceutically acceptable counterion, thiol, amine, carboxyl, and / or azide, preferably OH or said ammonium.

[0208] A preferred ammonium group is -N+(CH3)2(alkyl), wherein the alkyl comprises between 1 and 20 carbon atoms, preferably between 1 and 8 carbon atoms, and most preferably 2 carbon atoms. The ammonium group is preferably accompanied by a halogenide ion, preferably bromide (Br. Preferred said other pendant groups in repeat units (C) include: -CH2-CH2-OH and -CH2-CH2-N+(CH3)2(CH2CH3) Br-.

[0209] In any of the above embodiments, the BB polymer may further comprise one or more aforementioned “capping blocks”. A capping block is a functional group, a substituent, or a polymer or peptide attached at either or both ends of the backbone of the BB polymer. The nature of capping blocks will be chosen according to the properties to be imparted to the bottle-brush polymer. For example, capping blocks might be included to improve adhesion of the bottle-brush polymer to biological surfaces or biopolymers. Suitable capping blocks include halogen atoms (preferably Cl); alkyl, alkene or alkyne groups, the alkyl, alkene, and alkyne groups optionally being substituted or interrupted by one or more thiol, amine, carboxyl, and / or azide functional groups; peptides; as well as polymer chains bearing said functional groups or peptides.

[0210] It is well-known to skilled person that, when an initiator is used for polymerization, a remnant of such initiator (i.e. what is left of the initiator after reacting) typically becomes incorporated in the backbone of the polymer being polymerized. Remnants of monofunctional initiators are typically incorporated the end(s) of polymer backbone. Remnants of bifunctional initiators are typically incorporated between any two repeat units in the polymer backbone. Thus, the backbone of the BB polymer may comprise a remnant of an initiator molecule. Any remnant of any initiator molecule can be included. Non-limiting examples of such remnant include: -C(=O)-O-CH2-CH2-O-C(=O)-, which is a remnant of a bifunctional initiator and thus comprised between any two repeat units in the polymer backbone.

[0211] In preferred embodiments, the BB polymer comprises:

[0212] - repeat units (A) that are: , wherein x and R are as defined above,

[0213]

[0214] - repeat units (B) that are, and

[0215] - optionally repeat units (C) that are

[0216]

[0217] OH

[0218] In embodiments, the BB polymer comprises the following two repeat units: (A) preferably

[0219]

[0220] , and (B) I, wherein x and R are as defined above. In embodiments, the copolymer has a grafting ratio, between about 40% and about 60% (in other words, it comprises about 40% to about 60% of repeat unit (A), preferably it has a grafting ratio between about 45% and about 55%, and more preferably a grafting ratio of about 45% or about 55%. Preferably, this BB polymer is a statistical copolymer.

[0221] In preferred embodiments, the BB polymer comprises (A) preferably

[0222]

[0223] , wherein x and R are as defined above,

[0224] In preferred embodiments, the BB polymer is (PBiBEM-g-PMPC)-stat-PHEMA-stat-PMMA:

[0225]

[0226] preferably:

[0227]

[0228] preferably:

[0229] wherein x is as defined above and m and n represent repeat unit ratios (e.g., m = number of repeat units of the formula on the left. > - tota -l — num nb -er o;f - repeat uni —ts i —n po;l -ymer and, 0 < m < 1 and 0 ≤ n <1. Preferably, these BB

[0230]

[0231] polymers are statistical copolymers. In embodiments, these BB polymers have a grafting ratio between about 30% and about 100% (i.e., m varies from about 0.3 to about 1). In preferred embodiments, the grafting ratio is about 30%, about 35%, about 40%, about 45%, or about 50% or more and / or about 100%, about 90%, about 80%, about 70%, about 65%, about 60%, or about 55% or about 50% less. In more preferred embodiments, the grafting ratio varies from about 40% to about 60% ( / .e., m varies from about 0.4 to about 0.6), preferably about 45% to about 55% ( / .e., m varies from about 0.45 to about 0.55), more preferably about 40% to about 50% ( / .e., m varies from about 0.4 to about 0.5), and yet more preferably is about 45% or about 55% (m is about 0.45 or about 0.55). In embodiments, these BB polymers comprise a minor proportion of PHEMA, i.e., n is about 0.15, about 0.10, about 0.08, about 0.05, about 0.04, about 0.03, about 0.02, about 0.01, about 0.0075, about 0.005, or about 0.004 or less, preferably n is about 0.05 or less; or is about 0.05 or about 0.004. In preferred embodiments, the BB polymer is of formula (PBiBEM54o-g-PMPC28)-stat-PHEMA6o-stat-PMMA6oo or (PBiBEM456-g-PMPC35)-stat-PHEMA3-stat-PMMA37o, wherein the numbers represent the number of repeat units in the polymer i.e., in these polymers m=540 / 1200=0.45 and n=60 / 1200=0.05 and m=456 / 829=0.55 and n=3 / 829=0.0036, respectively).

[0232] In alternative embodiments, the BB polymer is said triblock copolymer, comprising a first block, a second block and a third block, the second block being located between the first block and the third block, the first block and the third block comprising statistically arranged repeat units B and C, while the second block comprises statistically arranged repeat units A and B. Preferably, said triblock copolymer comprises the following repeat units: (A)

[0233] O'^^O

[0234] O^ / 0

[0235] \x

[0236] o ’

[0237] O x / R

[0238]

[0239] °, preferably wherein x and R are as defined above, (B)

[0240]

[0241] In preferred embodiments, the BB polymer is of formula:

[0242]

[0243] , preferably of formula:

[0244]

[0245] , wherein x is as defined above, and wherein a, b, c, d, e, and f are ≥ 1 and represent the number of repeat units, preferably:

[0246] • a varies between about 25 and about 500, preferably between about 50 and about 200, more preferably between 50 and 100,

[0247] • b varies between about 25 and about 500, preferably between about 50 and about 200, more preferably between 100 and 150,

[0248] • c varies between about 100 and about 1000, preferably between about 200 and about 800, preferably between about 400 and about 500,

[0249] • d varies between about 100 and about 450, preferably between about 200 and about 800, preferably between about 350 and about 500,

[0250] • e varies between about 25 and about 500, preferably between about 50 and about 200, more preferably between 100 and 150, and

[0251] • f varies about 25 and about 500, preferably between about 50 and about 200, more preferably between 50 and 100;

[0252] more preferably, this BB polymer has a grafting ratio between about 25% and about 75%,

[0253] and most preferably of formula:

[0254]

[0255] , (which can be prepared as reported in Banquy et al., J. Am. Chem. Soc. 2014, 136, 17, 6199-6202).

[0256] In embodiments, the BB polymer, and particularly the pendant chains, does not comprise HA.

[0257] In embodiments, the concentration of BB polymer is about 1, about 25, about 50, about 75, about 85, about 90, about 95, about 100, about 150, about 200, about 250, about 300, about 350, about 400, about 450, or about 500 pg / ml or more and / or about 30, 25, 20, 15, 10, about 5, about 1, about 0.5, about 0.25, or about 0.1 mg / mL or less.

[0258] In an embodiment, the composition is free of any other polymer, for example any linear polymer. In another embodiment, the composition further comprises a linear polymer.

[0259] The linear polymer is a polymer that, contrary to the BB polymer, has a linear structure. It is thus free from polymeric pendant chains.

[0260] In embodiments, the linear polymer has a molecular weight of about 5 kDa, about 10 kDa, about 25 kDa, about 50 kDa, about 100 kDa, about 250 kDa, or about 500 kDa or more and / or 20 Mda, about 8 Mda, or about 5 Mda or less. In embodiments, the linear polymer has a molecular weight of about 10 kDa, about 20 Mda, about 500 kDa, or about 50 kDa.

[0261] In embodiments, the linear polymer is:

[0262] - O-CH2

[0263] < °

[0264] OHX - a-1,6

[0265] +

[0266] a-1,3

[0267] • dextran:

[0268]

[0269] poly(vinylpyrrolidone):

[0270]

[0271] polyethylene glycol):

[0272]

[0273] • hydroxypropyl cellulose,

[0274] • a polymethacrylate polymer or copolymer,

[0275] • a polyoxazoline polymer, or

[0276] • a polyacrylate polymer or copolymer,

[0277] wherein m and n represent the number of repeat units, or a (preferably pharmaceutically acceptable) salt thereof. In preferred embodiments, the number of repeat units is such that the linear polymer has the above-mentioned molecular weights. In embodiments, the linear polymer is partially crosslinked to achieve these molecular weights.

[0278] In other preferred embodiments, the linear polymer is poly(vinylpyrrolidone).

[0279] In embodiments, the concentration of the linear polymer in the composition is about 0.001 mg / mL, 0.005 mg / mL, 0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, or about 5 mg / mL or more. In preferred embodiments, the concentration of the linear polymer in the composition is about 0.01 mg I mL or more, and more preferably about 0.1 mg / mL or more, yet more preferably about 1 mg / mL or more, and most preferably about 0.9 mg / mL or about 2 mg / mL.

[0280] In embodiments, the composition comprises the BB polymer and the linear polymer in a BB polymer: linear polymer weight ratio between about 1:1 and about 1:20, preferably between about 1:2.5 and about 1: 15, more preferably between about 1:5 and about 1:10.

[0281] In embodiments, the solvent is saline ( / .e., an aqueous solution of mainly NaCI), preferably isotonic, preferably buffered at a pH of about 7 to about 7.4 using for example a phosphate buffer. In embodiments, the solvent is phosphate-buffered saline (PBS).

[0282] The composition may further comprise one or more pharmaceutically acceptable additives or excipients, such as for example inorganic or organic buffers (e.g., sodium or potassium salts of phosphate, carbonate, acetate, or citrate) and pH adjustment agents (e.g., hydrochloric acid, sodium or potassium hydroxide, salts of citrate or acetate, amino acids and their salts) antioxidants (e.g., ascorbic acid, alpha-tocopherol), surfactants (e.g., polysorbate 20, polysorbate 80, polyoxyethylene9- 10 nonyl phenol, sodium desoxycholate), solution and / or cryo / lyo stabilizers (e.g., sucrose, lactose, mannitol, trehalose), osmotic adjustment agents (e.g., salts or sugars), antibacterial agents (e.g., benzoic acid, phenol, gentamicin), antifoaming agents (e.g., polydimethylsilozone), preservatives (e.g., thimerosal, 2-phenoxy ethanol, EDTA), polymeric stabilizers and viscosity-adjustment agents (e.g., polyvinylpyrrolidone, poloxamer 488, carboxymethylcellulose) and co-solvents (e.g., glycerol, polyethylene glycol, ethanol). A composition formulated for injectable use may be in the form of a sterile saline solution for injection together with a preservative. Herein, “pharmaceutically acceptable” means generally accepted for use in pharmaceutical products. Of note, as is well known to the skilled person, whether a product is pharmaceutically acceptable depends on the end use of the composition. For example, components that may be acceptable in a composition intended to be applied topically may not be acceptable when the fluid is intended to be administered by injection.

[0283] The composition may further comprise one or more additional therapeutic agents, or the composition may be co-administered with one or more additional therapeutic agents. Such therapeutic agents may be chosen according to the end use of the composition. In an embodiment, the one or more additional therapeutic agents comprise anti-inflammatory agents and / or analgesics.

[0284] In one embodiment, the subject has arthritis such as osteoarthritis (OA), rheumatoid arthritis, acute gouty arthritis, psoriatic arthritis, reactive arthritis, arthritis due to Ehlers-Danlos Syndrome, haemochromatosis, hepatitis, Lyme disease, Sjogren's disease, Hashimoto's thyroiditis, celiac disease, non-celiac gluten sensitivity, inflammatory bowel disease, Henoch-Schbnlein purpura, Hyperimmunoglobulinemia D with recurrent fever, sarcoidosis, Whipple's disease, TNF receptor associated periodic syndrome, Granulomatosis with polyangiitis, familial Mediterranean fever, or systemic lupus erythematosus.

[0285] Kellgren and Lawrence classification system has been vastly used for classification of severity of osteoarthritis. Below is the original description: grade 0 (none): definite absence of x-ray changes of osteoarthritis;

[0286] grade 1 (doubtful): doubtful narrowing of joint space and possible osteophytic lipping; grade 2 (minimal): definite osteophytes and possible narrowing of joint space on posteroanterior weight-bearing radiograph;

[0287] grade 3 (moderate): moderate multiple osteophytes, definite narrowing of joint space and some sclerosis and possible deformity of bone ends;

[0288] grade 4 (severe): large osteophytes, marked narrowing of joint space, severe sclerosis and definite deformity of bone ends.

[0289] In an embodiment, the subject has grade 1 OA. In another embodiment, the subject has grade 2 OA. In another embodiment, the subject has grade 3 OA. In another embodiment, the subject has grade 4 OA.

[0290] The composition described herein can be used in combination with any of a variety of additional chemical entities, including but not limited to, analgesics such as local anesthetics (e.g., bupivacaine, ropivacaine, or lidocaine), and / or anti-inflammatory agents such as any NSAID or glucocorticoid (e.g., triamcinolone, methylprednisolone), or antioxidants (e.g., mannitol, sorbitol), or biologies such as stem cells, platelet-rich plasma or derived protein solutions. In some embodiments, the composition described herein and additional chemical entities are formulated into a single therapeutic composition, and the composition and the additional chemical entities are administered simultaneously. Alternatively, the composition described herein and the additional chemical entities are separate from each other, e.g., each is formulated into a separate therapeutic composition, and the composition and the additional chemical entities are administered simultaneously, or at different times during a treatment regimen by the same route or different routes, as a single dose or multiple doses.

[0291] EXAMPLES

[0292] The present disclosure is illustrated in further details by the following non-limiting examples.

[0293] Example 1: Safety study in healthy rats (Phase 1)

[0294] Preparation of the formulations

[0295] The bottle-brush polymer used in the present studies has the following structure:

[0296]

[0297] It was prepared according to the method reported in Banquy et al., J. Am. Chem. Soc. 2014, 136, 17, 6199-6202).

[0298] For the Phase 1, safety study in healthy Sprague-Dawley rats, three doses (1X, 3X and 5X) were tested for each of the three products: Bottle-brush polymer (BB), hyaluronic acid (HA) and BB-HA association. For the BB group, the injection volume was 30.0 pL for the 1X (0.2 mg) with a solution concentration of 6.7 mg / mL and for the 3X (0.6 mg) with a solution concentration of 20.0 mg / mL. For the 5X (1.0 mg) group, the injection volume was 40.0 pL with a solution concentration of 25.0 mg / mL. For the HA and BB-HA groups, the injection volumes were 14.0 pL for the 1X dose, 40.0 pL for the 3X and 65.0 pL for the 5X with a solution concentration of 15.0 mg / mL. All injections were performed with previously sterilized Hamilton™ syringes with a total volume of 250.0 pL and dead volume of 50.0 pL considered during injection. Each syringe was mounted with a 25G x 1 inch needle.

[0299] The Phase 2 focused on the efficacy of the three products in osteoarthritic Sprague-Dawley rats (validated MI-RAT model) in comparison with a placebo (0.9% saline solution). The three products were tested at a dose of 2X (0.4 mg) and the injection volume was 26.6 pL per rat (solution concentration of 15.0 mg / mL). Each injection was contained in pre-filled 1.0 mL syringe with a 25G x 5 / 8 inch needle.

[0300] The Phase 3, which studied the safety of BB in healthy Beagle dogs, was carried out in comparison with a naive dog (0.9% saline solution). Two doses were tested, 2X (16.0 mg) and 4X (32.0 mg), and injected through a 3.0 mL pre-filled syringe containing 1.0 mL of product and mounted on a 20G x 1 inch needle.

[0301] For each phase, there was no other addition of excipient or solute in the injectate, and all syringes were kept in the refrigerator (4°C) then brought back to room temperature (21 °C) 20 minutes before injection.

[0302] Table 1: Experimental design of a prospective, randomized, dose-controlled, blinded study Sample size (N =

[0303] Treatment (BSL Haemato- Veterinary 45; ovariectomized

[0304] Group intraarticular biochemical clinical Sprague-Dawley

[0305] stifle injection) evaluation examination rats; 200-300g)

[0306] HA 1X n = 5 0.2 mg

[0307] HA 3X n = 5 0.6 mg

[0308] HA 5X n = 5 1.0 mg

[0309] BB 1X n = 5 0.2 mg

[0310] BB 3X n = 5 0.6 mg

[0311] BB 5X n = 5 1.0 mg BSL and

[0312] D60 BB-HA 0.2 mg BB + 0.2 D59

[0313] n = 5

[0314] 1X mg HA

[0315] BB-HA 0.6 mg BB + 0.6

[0316] n = 5

[0317] 3X mg HA

[0318] BB-HA 1.0 mg BB + 1.0

[0319] n = 5

[0320]

[0321] 5X mg HA

[0322] HA: Hyaluronic acid; BB: Bottle-brush polymer; D: Day; BSL: Baseline

[0323] Outcome measurement

[0324] • General behavior (daily) and body weight (weekly);

[0325] • Veterinary clinical examination (SOAP);

[0326] • Standard haemato-biochemical evaluation;

[0327] • Macro- and histo-morphometric post-mortem analysis of stifles (with OARSI scoring scale Gewin., et al. 2010) and vital organs (liver, spleen and kidney), by 2 independent and blinded observers: Ph. D. student and pathologist.

[0328] Results

[0329] • No side effect observed;

[0330] • Histological analysis of the stifle: OARSI score of zero;

[0331] • Pathological analysis of organs: no lesion and / or repair, with absent foreign elements.

[0332] Conclusion The results of Phase 1 allowed to state positively about the safety of tested products (BB, HA and BB-HA) and / or the different doses (1X, 3X and 5X) injected into the stifle joint space of healthy Sprague-Dawley rats.

[0333] Example 2: Safety study in healthy dogs (Phase 3)

[0334] Table 2: Experimental design of a prospective, randomized, dose-controlled, blinded study Sample size (N

[0335] Veterinary clinical and = 9; non- Treatment (BSL Haemato- orthopedic Group ovariectomized intraarticular biochemical

[0336] examination Beagle dogs; stifle injection) evaluation

[0337] (by 2 veterinarians) 6-9kg)

[0338] BB 2X n = 4 16 mg BSL; D20*; D41* BSL; D6*; D20*; D41* BB 4X n = 4 32 mg BSL; D20; D41 BSL; D6; D20; D41 Saline 0.9% BSL; D20; D41 BSL; D6; D20; D41 NAIVE n = 1

[0339]

[0340] solution

[0341] BB: Bottle-brush polymer; D: Day; BSL: Baseline; *Pilot dog (n = 1; BB 2X)

[0342] Outcome measurement

[0343] • General behavior (daily) and body weight (weekly);

[0344] • Veterinary clinical examination (SOAP);

[0345] • Orthopaedic examination of both stifles (daily for 1 week after injection, then weekly) by Ph. D. student and / or veterinarians;

[0346] • Standard haemato-biochemical evaluation;

[0347] Results

[0348] • No side effect observed;

[0349] • Haemato-biochemical parameters: Glycaemia slight and punctual decrease in n=5 dogs, Blood Urea Nitrogen slight increase and Creatinine slight decrease in quasi- all dogs, associated with hematocrit and total proteins in high range normal values. These “abnormalities” reflected changes due to dehydration (summer) with short water and food fasting before possible sedation. All other outcomes were in normal ranges;

[0350] Conclusion

[0351] The results of Phase 3 allowed to state positively about the BB safety and / or the different doses (2X and 4X) injected into the stifle joint space of healthy Beagle dogs.

[0352] Example 3: Efficacy study in osteoarthritic (OA) rats (Phase 2) Table 3: Experimental design of a prospective, randomized, blinded, comparative (to negative control) study

[0353] Sample size (N

[0354] Treatment

[0355] = 60;

[0356] (D11 Euthanasia ovariectomized Surgey Phenotypic

[0357] Group intraarticular and Sprague- (BSL) evaluations

[0358] injection) Sampling Dawley rats;

[0359] 200-300g)

[0360] NAIVE n = 12 - - MI-RAT CCLT-DMM Saline 0.9%

[0361] n = 12 n = 12 / gr.: BSL;

[0362] Placebo (DO) solution

[0363] D42 MI-RAT CCLT-DMM D21; D41;

[0364] n = 12 0.2 mg BB (n = 6 / gr.) BB 2X (DO)

[0365] n = 6 / gr.: D61; and MI-RAT CCLT-DMM

[0366] n = 12 0.2 mg HA D84 HA 2X (DO) D83

[0367] (n = 6 / gr.) MI-RAT

[0368] CCLT-DMM 0.2 mg BB +

[0369] BB-HA n = 12

[0370] (DO) 0.2 mg HA

[0371] 2X

[0372]

[0373] MI-RAT: Montreal Induction o Rat Arthritis Testing model; HA: Hyaluronic acid; BB: Bottlebrush polymer; CCLT-DMM: Transection of the cranial cruciate ligament with destabilization of the medial meniscus; D: Day; BSL: Baseline

[0374] Outcome measurement

[0375] • General behavior (daily) and body weight (weekly);

[0376] • Macro- and histo-morphometric post-mortem analysis of stifles (with OARSI scoring) by 2 independent and blinded observers: Ph. D. student and pathologist.

[0377] • Phenotypic assessment of or pain: Static weight-bearing (SWB) of both hindlimbs, Punctate tactile paw withdrawal threshold (PWT) of both hind-paws;

[0378] • Dynamic evaluation for pain system: Response to mechanical temporal summation (RMTS) at the tail level; Conditioned pain modulation (CPM) with PWT, i.e. pain inhibition with PWT post-CPM measured after post-conditioning stimulus (ear clip) placement, and pain facilitation (or sensitization) with PWT post-RMTS.

[0379] Results

[0380] No side effect observed.

[0381] A) Histo-morphometric analysis of OA-afflicted stifles

[0382] Frontal section (depth = 2000pm) of stifle (n=4 stifles / group (n=5) / time (n=2), except n=1 for MI-RAT Placebo on D84) had been colored with hematoxylin and eosin (H& E) and safranin-O staining. Different sections of stifle were evaluated by 2 evaluators separately, Ph. D. student and pathologist, via OARSI scoring scale (Gewin., et al. 2010) and OA damage safranin-0 (Glasson., et al. 2010). Each criterion was evaluated by mixed model.

[0383] 1 - Total histological OARSI score.

[0384] The results depicted in FIG. 1 show:

[0385] • Significant increase in total OARSI score for MI-RAT groups at each time point relative to the Naive group;

[0386] • Significant increase by 51% in total OARSI score over time (from D42 to D84) for MI-RAT Placebo (P= 0.016);

[0387] • At D42: Significant difference of MI-RAT BB with MI-RAT Placebo (b vs. c; P = 0.017), or with MI-RAT BB-HA (b vs. c; P= 0.047);

[0388] • At D84: Significant difference between MI-RAT Placebo and the 3 treatment groups (b vs. c; P< 0.005).

[0389] 2 - Cartilage degeneration score.

[0390] The results depicted in FIG. 2 show:

[0391] • Significant increase in cartilage degeneration for MI-RAT groups at each time point relative to the Naive group;

[0392] • At D42: Significant difference of MI-RAT BB with MI-RAT Placebo (b vs. c; P = 0.004), or with MI-RAT BB-HA (b vs. c; P= 0.018);

[0393] • At D84: Significant difference of MI-RAT Placebo with MI-RAT BB (b vs. c; P = 0.039), or with MI-RAT BB-HA (b vs. c; P= 0.005).

[0394] 3 - Osteophyte score.

[0395] The results depicted in FIG. 3 show:

[0396] • At D42: No detection of osteophyte, regardless of the group;

[0397] • At D84: Significant increase for MI-RAT Placebo and MI-RAT BB-HA (P = 0.003 for both groups between D42 and D84 values); Significant difference of both latter groups with MI-RAT BB and MI-RAT HA with an amplitude of +86% in the score (a vs. b; P< 0.012).

[0398] 4 - Calcified cartilage and subchondral bone damage score.

[0399] The results depicted in FIG. 4 show:

[0400] • At D42: Significant increase in calcified cartilage and subchondral bone damage score for MI-RAT Placebo relative to Naive, MI-RAT BB and MI-RAT HA (P< 0.034); At D84: Significant increase in MI-RAT Placebo relative to the Naive group (P = 0.036).

[0401] 5 - Synovitis score.

[0402] The results depicted in FIG. 5 show:

[0403] • At D42: Significant increase in synovitis score for MI-RAT BB-HA relative to Naive (P= 0.031);

[0404] • At D84: Significant increase in synovitis score (+63%) of MI-RAT Placebo relative to D42 (P = 0.044). The synovitis score of the Naive, MI-RAT BB and MI-RAT HA groups was significantly lower than the synovitis score of MI-RAT Placebo (P < 0.045), when the MI-RAT BB-HA was not lower than the latter, and was higher in comparison to the Naive group (a vs. b,c; P= 0.031).

[0405] 6 - Loss of proteoglycans score.

[0406] The results depicted in FIG. 6 show:

[0407] • At D42 and D84: Significant increase in the loss of proteoglycans (safranin-0 staining) between Naive and all MI-RAT groups (a vs. b; P< 0.05), except MI-RAT BB at D84;

[0408] • Increase overtime for MI-RAT Placebo by +46% between D42 and D84 (P= 0.009);

[0409] • At D84: Significant difference between MI-RAT Placebo and all treatment groups:

[0410] MI-RAT BB (a vs. b; P= 0.006); MI-RAT HA and MI-RAT BB-HA (b vs. c; P< 0.036); and between Naive and MI-RAT Placebo, HA and BB-HA groups (a vs. b / c; P < 0.006).

[0411] B) Descriptive macroscopic analysis of OA-afflicted stifles

[0412] Dissection and Evans’s blue staining of stifle (n=2 stifles / group (n=5) / time (n=2)) were photographed for 4 compartments (medial and lateral femoral condyles, as well as medial and lateral tibial plateaus) (Otis., et al. 2017). Quantification, by 2 blinded observers, on Imaged of macroscopic cartilage lesions: descriptive data of the percentage of surface alteration and lesion severity index via a Likert scale from 0 to 4 (0 = normal; 4 = maximum severity) (Guingamp., et al. 1997). Due to the low sample size, the analysis was only about descriptive data.

[0413] The results depicted in FIGs. 7A-B and 8 show:

[0414] • At D42: MI-RAT groups have an altered surface area and a severity index 4x higher than the Naive group. No apparent difference between the four MI-RAT Placebo and treated groups; • At D84: Difference between MI-RAT groups, with a reduction in the surface alteration and severity index for the MI-RAT BB group in first, (-44.0% reduction in surface alteration vs. MI-RAT Placebo), then MI-RAT BB-HA (-27.1% reduction in surface alteration vs. MI-RAT Placebo) and MI-RAT HA (-21.8% reduction vs. MI- RAT Placebo). Finally, the severity index reached a peak of 10 (max = 16 for the four compartments) in MI-RAT Placebo at D84, and decreased by -45.0% in MI-RAT BB, and -35.0% in MI-RAT BB-HA, where there was no decrease in MI-RAT HA (index of 9.8).

[0415] Conclusions on histology data

[0416] On histology, the three intra-articular treatments applied 11 days after OA induction in the right stifle were beneficial to reduce the degree of structural damages. The difference was obvious at D84, indicating a delay is necessary for the osteoarthritic lesions to be well established in the MI-RAT model (confirming the face validity of the MI-RAT model as representative of real osteoarthritic lesions). Moreover, clearly the MI-RAT BB group was the most constant in structural benefits, and the only group to present significant differences, relative to MI-RAT Placebo, as soon as D42. This was particularly true for Total histological score, Cartilage degeneration and Calcified cartilage and subchondral bone damages. For Osteophyte, Synovitis, and Loss of proteoglycans, it was necessary to wait up to D84 for the MI-RAT BB to show significant difference relative to MI-RAT Placebo. Interestingly, for these three outcomes, the MI-RAT BB group was similar to the Naive group, when MI-RAT HA was for Osteophyte and Synovitis, and MI-RAT BB-HA was always worse (higher) than Naive. The preservative effect of loss of proteoglycans induced only by MI-RAT BB supports a major site of action for BB at the hyaline cartilage level (replacing loss of natural proteoglycans such as chondroitin-6-sulfate, keratan sulfate, or lubricin - also called proteoglycan 4, PRG4, or superficial zone protein -). The two other treatments, i.e. HA and BB-HA, were more erratic in structural benefits. The structural benefits recorded on histology for the MI-RAT BB group were translated in the macroscopic morphometric assessment, where only the MI-RAT BB presented a -45% reduction in both surface alteration and severity index.

[0417] C) Pain phenotype and pain facilitation / modulation

[0418] At baseline (BSL, n = 60 rats), the measurements were highly reproducible and homogenous, and no disequilibrium was observed between left and right hind limbs either for SWB (spontaneous pain expression on weight-bearing forces) or PWT (evoked pain withdrawal threshold) (Table 4).

[0419] Table 4 SWB Baseline

[0420] % asymmetry R on L

[0421] LHP (%BW) RHP (%BW)

[0422] Mean 38.7 38.9 0.42%

[0423] SD 2.7 2.8

[0424] PWT Baseline % asymmetry R on L

[0425] LHP (%BW) RHP (%BW)

[0426] Mean 20.3 19.7 -3.01%

[0427] SD 4.1 4.4

[0428] The cut-off number of RMTS stimulations was set at 30 (after validation), and a huge majority of rats reached this value at BSL, when no pain was present (Table 5).

[0429] Table 5

[0430] RMTS 27.3 (4.5) Mean (SD) Baseline 29.5 Median

[0431] Finally, the PWT post-RMTS must be decreased, compared to PWT, as a marker of nociceptive

[0432] facilitation induction, whereas the PWT post-CPM must be increased, compared to PWT, as a marker of nociceptive inhibitory endogenous control.

[0433] This was the case as the median decrease in PWT post-RMTS was -22.5% in Left Hind Paw, and -23.2% for Right Hind Paw, when the median increase in PWT post-CPM was +29.0% in Left Hind Paw and +30.0% in Right Hind Paw. Higher is the change, more active is the facilitation / inhibition process, and as expected, only about 80% of rats were responders to either RMTS-facilitation or CPM-inhibition, at BSL.

[0434] a) Spontaneous expression of pain (SWB)

[0435] The results depicted in FIG. 9 show:

[0436] • Type III fixed effects: Group (P < 0.001), Time (P < 0.001) and Group*Time interaction (P< 0.001);

[0437] • Post-OA induction: All MI-RAT groups had a decrease at D22 in SWB asymmetry index compared to BSL

[0438] o more significant for MI-RAT Placebo group (-75%; P = 0.003),

[0439] o less significant for MI-RAT BB at first (-35%; P = 0.028), o then MI-RAT BB-HA (-45%; P = 0.001) and MI-RAT HA (-55%; P = 0.002);

[0440] • MI-RAT BB: Significant difference with MI-RAT BB-HA (at D41, P = 0.042; at D61, P = 0.012 and at D83, P = 0.034); and MI-RAT HA (at D83, P = 0.034). Only MI- RAT BB was not different with Naive, unlike the 3 other groups, at D61 (P = 0.022) and D83 (P = 0.029). In MI-RAT BB, SWB D22 and D41 were different from D61 and D83 (P< 0.024).

[0441] b) Evoked expression of pain (PWT)

[0442] The results depicted in FIG. 10 show:

[0443] • Type III fixed effects: Group (P < 0.001), Time (P < 0.001) and Group*Time interaction (P< 0.001). Only the Time effect remains when testing 2 by 2 groups;

[0444] • Post-OA induction: All MI-RAT groups had a decrease at D22 in PWT asymmetry index compared to BSL

[0445] o more significant for MI-RAT Placebo group (-60%; P= 0.001), o less significant for MI-RAT BB at first (-35%; P = 0.007),

[0446] o then MI-RAT BB-HA (-40%; P = 0.001) and MI-RAT HA (-55%; P = 0.024) • At D22: Significant difference in PWT decrease between MI-RAT Placebo and MI- RAT BB (P = 0.009) and BB-HA (P = 0.040) groups;

[0447] • At D41 and D61: No significant difference;

[0448] • At D83: Significant difference between MI-RAT BB-HA and MI-RAT BB (P= 0.011), or Naive (P = 0.002). For MI-RAT BB, at D83, PWT was different from the other previous timepoints (P< 0.007), except BSL.

[0449] c) Pain central sensitization (RMTS)

[0450] The results depicted in FIG. 11 show:

[0451] • Type III fixed effects: Group (P < 0.001), Time (P < 0.001) and Group*Time interaction (P< 0.003). Only the Time effect remains when testing 2 by 2 groups;

[0452] • Post-OA induction: All MI-RAT groups had a similar decrease in the number of stimulations, by almost -50% compared to the Naive group (P < 0.020) and their BSL value (P< 0.013);

[0453] • No significant difference between the four Placebo and treated groups at each time and no Placebo or treated group returned to its BSL value.

[0454] Pain

[0455]

[0456] inhibition

[0457]

[0458] The results depicted in FIG. 12A (right hind paw, on which stifle OA was induced) show:

[0459] • Type III fixed effects: Group (P< 0.009) and Time (P= 0.065); • At D22: Highly functional CPM for MI-RAT Placebo (+107%; P = 0.001) and MI-RAT HA (+55%; P = 0.019), which passed out overtime;

[0460] • MI-RAT BB and BB-HA groups seemed to not solicit their inhibitory endogenous control of pain.

[0461] The results depicted in FIG. 12B (left hind paw) show:

[0462] • Type III fixed effects: no significance;

[0463] • At D22: Very functional CPM for MI-RAT Placebo (+57%; P = 0.046) which passed out over time and significant difference between MI-RAT Placebo and MI-RAT BB & BB- HA at D22 (P < 0.026), and close to significance between MI-RAT Placebo (whom CPM functionality declined) and MI-RAT BB (P = 0.068) and MI-RAT HA (P = 0.064) at D83.

[0464]

[0465] Pain facilitation

[0466] The results depicted in FIG. 13A (right hind paw, source of primary pain) show:

[0467] • Type III fixed effects: Time (P = 0.001);

[0468] • Decrease in PWT post-RMTS of RHP for all groups at each time (from 0% to -50%);

[0469] a clear tendency to decrease with-time (meaning more central facilitation or sensitization) is observed on the graph for all MI-RAT groups but MI-RAT BB, with the exception of one peak for MI-RAT Placebo at D22, and fluctuant variations in different groups;

[0470] • At D22: Significant difference between MI-RAT Placebo and MI-RAT HA (P= 0.017) and MI-RAT BB-HA (P = 0.037), the two latter being more sensitized;

[0471] • At D83: Significant difference between Naive and MI-RAT Placebo (P= 0.049) and MIRAT HA (P= 0.009), the two latter being more sensitized;

[0472] • MI-RAT BB values followed the same trend as Naive group;

[0473] • MI-RAT HA: Most negatively impacted with a drop by -40% over time (BSL vs. D22;

[0474] P = 0.027 and BSL vs. D83; P < 0.001);

[0475] • MI-RAT BB-HA: Significant difference between BSL and D22 (P= 0.030) and D83 (P = 0.033)

[0476] The results depicted in FIG. 13B (left hind paw, source of secondary pain) show:

[0477] • Type III fixed effects: Time (P = 0.011);

[0478] • Decrease in PWT post-RMTS of LHP for all groups at each time (from -20% to - 45%) and particularly for MI-RAT BB-HA at D22, and MI-RAT HA from D22 to D83, i.e. a decrease by -15% compared to BSL values;

[0479] • MI-RAT BB group seems to have stable values over time (-25%), similar to BSL values; • MI-RAT HA: Negatively impacted with a drop by -40% over time (BSL vs. D83; a vs. b; P = 0.021) and significant difference at D83 with MI-RAT BB and BB-HA (P < 0.039);

[0480] • MI-RAT BB-HA: Significant difference between D22 and BSL (b vs. a; P = 0.010) and D83 (b vs. c; P = 0.025).

[0481] Conclusions on pain data

[0482] On the phenotypic assessment of pain and dynamic evaluation of pain processes, the result outweighed expectations.

[0483] • For SWB (spontaneous pain behavior) and PWT (evoked pain behavior), clearly MI-RAT BB showed lower (subacute, D22) deterioration than MI-RAT Placebo, and better recovery than other treatments applied to MI-RAT (Placebo, HA, and BB-HA).

[0484] • RMTS: The OA induction generated a central sensitization that was not sensitive to distinguish between Placebo and ‘treated’ groups. The term ‘treatment’ is relative as all tested compounds are lubricating inert solutions (with an expected disease-OA- modifying drug (DMOAD) effect) and not supposed to interfere with the pathologic pain process. As soon as D22, RMTS reached a nadir and was not really modified subsequently in any MI-RAT group.

[0485] • PWT post-CPM: The MI-RAT Placebo and HA groups solicited their inhibitory endogenous control of pain, most importantly at D22 (for Placebo) on both sides, RHP and LHP, indicative of a central nervous system intervention. After the peak in pain inhibition solicitation observed at D22, the needs in pain inhibition decreased, and it is possible the MI-RAT Placebo to show some fatigue in the endogenous inhibition of pain at D83 (LHP mostly). At the same time, the MI-RAT BB and BB-HA groups seemed to not solicit their inhibitory endogenous control of pain.

[0486] • PWT post-RMTS: Clearly, the MI-RAT Placebo and both HA groups were more sensitized, with BB-HA intermediate, and BB responding similar to the Naive group. The fact that the MI-RAT Placebo was not responsive at LHP side, but presented a peak of inhibiting facilitation at D22 for RHP could be explained by the higher solicitation of the inhibitory endogenous control (see above PW post-CPM) at the same timepoint, which would counteract the pain facilitation process centrally.

[0487] Overall, the different phenotypic evaluations of pain as well as the histological and macroscopic analyses of stifle associated an attractive structure-function benefit induced by the BB solution. The latter was superior in most aspects to HA, when BB-HA group showed some intermediate response, indicative that HA interferes with the benefits induced by BB. The BB compound appears to protect the stifle articular structures: its boundary lubrication ability could be associated with some matrix-binding capacity leading to a clear DMOAD effect. The latter was translated in functional benefits on both direct pain (either spontaneous or evoked) expression and on nociceptive process of facilitation / inhibitory endogenous control, associated to no solicitation of either facilitation or inhibition process which could be translated on long-term to a better preservation of the central nervous system plasticity. Finally, the BB solution looks to be safe with intra-articular single administration both in rats and in dogs.

[0488] Example 4: Effects of the treatment with HA, BB and BB-HA on the neuropeptidome The effects of the treatment with HA, BB and BB-HA on the neuropeptidome (peptides present in spinal cord) was assessed using the method described in Lu, G et al. Global spinal cord peptidome profiling in response to osteoarthritis in rats. Molecular Omics. 21, 645-656, October 2025. The method was applied on spinal cord samples from OA rats collected at day 42 (D42) and day 84 (D84) during the above-described experiments. FIGs. 14A-B present the Principal Component Analysis (PCA) of the whole spinal neuropeptidome measured in rats associated with the three groups of treatment, i.e. Hyaluronic acid (HA) as positive control; “Bottle-Brush” (BB) polymer; and their combination (BBHA).

[0489] The measurement of spinal neuropeptidome shows that:

[0490] • At D42 (FIG. 14A): BB has a striking different expression of neuropeptidome from the two other groups, but it is important to note that both HA and BB presented, at this timepoint (so 31 days after the respective treatment intra-articular injection at D11), a more restrained distribution of neuropeptidome than BBHA.

[0491] • At D84 (FIG. 14B): there was more overlap among the three clusters, but the most important result is that BB remained with a restrained distribution of neuropeptidome than both HA and BBHA.

[0492] This supports the following interpretation:

[0493] 1) HA, as reference control of viscosupplement, was able to induce some effect on the spinal cord release of neuropeptides, as observed at D42, but this effect was no longer observed at D84.

[0494] 2) BB presented a striking effect on spinal neuropeptidome at both D42 and D84, supporting the beneficial effects described on the pain phenotype (Example 3). 3) BBHA did not correspond to a combination of BB and HA. Indeed, the generated mixture acted differently of both constitutive compounds, at both D42 and D84, and induced a large reaction in the neuropeptidome, supporting the reaction of the central nervous system to fight the induced OA pain.

[0495] Overall, the spinal neuropeptidome data provides evidence that BB injected intra-articularly was associated to protective effect on the central nervous system, supporting the view that it would result in a better preservation of the central nervous system plasticity over the long-term in OA affected patients.

[0496] Although the present invention has been described hereinabove by way of specific embodiments thereof, it can be modified, without departing from the spirit and nature of the subject invention as defined in the appended claims. In the claims, the word "comprising" is used as an open-ended term, substantially equivalent to the phrase "including, but not limited to". The singular forms "a", "an" and "the" include corresponding plural references unless the context clearly dictates otherwise.

Claims

WHAT IS CLAIMED IS:

1. A method for preventing or reducing joint damage and / or pain in a subject suffering from a joint disease or condition, the method comprising administering to the subject an effective amount of a composition comprising a bottle-brush (BB) polymer comprising a polymeric backbone made of (A) repeat units bearing a polymeric pendant chain and (B) repeat unit free of the polymeric pendant chain, wherein the composition is free of hyaluronic acid (HA) and is not co-administered with a composition comprising HA.

2. The method according to claim 1, wherein the backbone of the BB polymer is acrylate or methacrylate based.

3. The method according to claim 1 or 2, wherein the backbone of the BB polymer is poly(methacrylate) or a poly(alkyl methacrylate).

4. The method according to any one of claims 1 to 3, wherein the backbone of the BB polymer is poly(methyl methacrylate).

5. The method according to any one of claims 1 to 4, wherein the polymeric pendant chain is attached directly to the backbone of the BB polymer or attached via a linking group.

6. The method according to claim 5, wherein the polymeric pendant chain is attached to the backbone of the BB polymer via the linking group.

7. The method according to claim 5 or 6, wherein the linking group attaching the polymeric pendant chain to the backbone of the BB polymer is a carboxylic acid, an ester, an amine, an azide, or a thiol functional group, or an alkylene, alkenylene, or alkylene group that is interrupted or not with one or more ester, amine, or azide, or thiol functional group, preferably the linking group is *-CH2-CH2-O-C(=O)-#, wherein * is the point of attachment of the backbone and#is the point of attachment to the polymeric pendant chain.

8. The method according to any one of claims 1 to 7, wherein the polymeric pendant chain is zwitterionic.

9. The method according to any one of claims 1 to 8, wherein the polymeric pendant chains comprise repeat units (D) having optionally attached thereto a substituent, either directly or through a linking group.

10. The method according to claim 9, wherein repeat units (D) are repeat units of:• poly(acrylic acid), such as a poly(alkyl acrylic acid),• a poly(acrylate), such as a poly(alkyl acrylate), for example poly(methyl acrylate), • poly(methacrylic acid), such as poly(alkyl methacrylic acid), for example poly(methyl methacrylic acid),• a poly(methacrylate), such as a poly(alkyl methacrylate), for example poly(methyl methacrylate),• poly(acrylamide), or• a poly(methacrylamide), such as a poly(alkyl methacrylate), for example poly(methyl methacrylate)preferably repeat units of a poly(methacrylate).

11. The method according to claim 9 or 10, wherein repeat units (D) have attached thereto a substituent either directly or through the linking group.

12. The method according to any one of claims 9 to 11, wherein the substituent is attached to repeat units (D) through the linking group.

13. The method according to any one of claims 9 to 12, wherein the linking group attaching the substituent to the repeat units (D) is a carboxylic acid, an ester, an amine, an azide, or a thiol functional group, or an alkylene, alkenylene, or alkylene group that is interrupted or not with one or more ester, amine, or azide, or thiol functional group, preferably the linking group is - CH2-CH2-.

14. The method according to any one of claims 9 to 13, wherein the substituent attached to repeat units (D) is a phosphorylcholine group, a saccharide or disaccharide group including but not limited to glucose, sucrose, lactose and their derivatives, such as D-gluconolactone and lactobionolactone, or a biocompatible hydrophilic group such as hydroxy, oligo(ethylene oxide), carboxy, amino, sulfo, thiol, phosphate, or a derivative thereof.

15. The method according to any one of claims 9 to 14, wherein the substituent attached to repeat units (D) is phosphorylcholine.

16. The method according to any one of claims 1 to 15, wherein the polymeric pendant chain is poly(2-methacryloyloxyethyl phosphorylcholine) of formula:

17. The method according to any one of claims 1 to 16, wherein the BB polymer comprises repeat units of formula:wherein R is H, an alkyl, a halogen (e.g., Br, Cl), or a peptide (e.g., a peptide comprising one or more cysteine residues), andwherein x represents the number of repeat unit and preferably varies between 1 and 1000, more preferably between 10 and 500, yet more preferably between 20 and 250, and most preferably between 25 and 100.

18. The method according to any one of claims 1 to 17, wherein the BB polymer further comprises (C) repeat units bearing another pendant group.

19. The method according to claim 18, wherein said other pendant group in repeat units (C) is an alkylene, alkenylene, or alkynylene (preferably alkylene), each of which being optionally interrupted or terminated by one or more of OH, ammonium accompanied by a pharmaceutically acceptable counterion, thiol, amine, carboxyl, and / or azide, preferably OH or said ammonium.

20. The method according to claim 19, wherein the alkylene comprises between 1 and 20 carbon atoms, preferably between 1 and 8 carbon atoms, and most preferably 2 carbon atoms and wherein the alkenylene and alkynylene comprise between 2 and 20 carbon atoms, preferably between 2 and 8 carbon atoms, and most preferably 2 carbon atoms.

21. The method according to claim 19 or 20, wherein said other pendant groups in repeat units (C) is an alkylene, alkenylene, and alkynylene group (preferably alkylene), each of which being terminated by OH, ammonium accompanied by a pharmaceutically acceptable counterion, thiol, amine, carboxyl, and / or azide, preferably OH or said ammonium.

22. The method according to any one of claims 19 to 21, wherein said ammonium group is -N+(CH3)2(alkyl), wherein the alkyl comprises between 1 and 20 carbon atoms, preferably between 1 and 8 carbon atoms, and most preferably 2 carbon atoms; preferably the ammonium group is preferably accompanied by a halogenide ion, preferably bromide (Br).

23. The method according to any one of claims 19 to 22, wherein said other pendant groups in repeat units (C) is -CH2-CH2-OH and -CH2-CH2-N+(CH3)2(CH2CH3) Br.

24. The method according to any one of claims 19 to 23, wherein the BB polymer is a triblock copolymer comprising a first block, a second block and a third block, the second block being located between the first block and the third block, the first block and the third block comprising statistically arranged repeat units B and C, while the second block comprises statistically arranged repeat units A and B.

25. The method according to any one of claims 1 to 24, wherein the backbone of the BB polymer further comprises a remnant of an initiator molecule.

26. The method according to any one of claims 1 to 25, wherein the BB polymer further comprises one or more capping blocks.

27. The method according to any one of claims 1 to 26, wherein the BB polymer comprises:- repeat units (A) that are:, wherein x and R are as defined above,CHooptionally repeat units (C) that areOH28. The method according to any one of claims 1 to 27, wherein the BB polymer comprises the following repeat units:(A), preferablyand (B)wherein x and R are as defined above;preferably with a grafting ratio between about 40% and about 60%, more preferably between about 45% and about 55%, and most of about 45% or about 55%.

29. The method according to any one of claims 1 to 27, wherein the BB polymer comprises the following repeat units:(A) preferably O^'-'O, wherein x and R are as defined above,preferably the BB polymer is of formula:preferably:preferably:wherein x is as defined above and m and n represent repeat unit ratios (e.g., m = number of repeat units of the formula on the leftand, 0 < m < 1 and 0 ≤ n <1;total number of repeat units in polymerpreferably having a grafting ratio between about 30% and about 100%; more preferably the grafting ratio is about 30%, about 35%, about 40%, about 45%, or about 50% or more and / orabout 100%, about 90%, about 80%, about 70%, about 65%, about 60%, or about 55% or about 50% less.

30. The method according to any one of claims 1 to 28, wherein the BB polymer is (PBiBEM540-g-PMPC28)-stat-PHEMA60-stat-PMMA600or (PBiBEM456-g-PMPC35)-stat-PHEMA3-stat-PMMA370.

31. The method of claim 24, wherein the triblock copolymer comprises repeat units:(A) preferably, wherein x and R are as defined above,(B) preferably the BB polymer is of formula:wherein x and R are as defined above, and wherein a, b, c, d, e, and f are ≥ 1 and represent the number of repeat units,preferably:a varies between about 25 and about 500, preferably between about 50 and about 200, more preferably between 50 and 100,b varies between about 25 and about 500, preferably between about 50 and about 200, more preferably between 100 and 150,c varies between about 100 and about 1000, preferably between about 200 and about 800, preferably between about 400 and about 500,d varies between about 100 and about 450, preferably between about 200 and about 800, preferably between about 350 and about 500,e varies between about 25 and about 500, preferably between about 50 and about 200, more preferably between 100 and 150, andf varies about 25 and about 500, preferably between about 50 and about 200, more preferably between 50 and 100;more preferably the BB polymer has a grafting ratio between about 25% and about 75%, and most preferably the BB polymer is of formula:b Br'N obBr32. The method according to any one of claims 1 to 30, wherein the molecular weight of the backbone is about 10 kDa, about 20 kDa, about 30 kDa, about 40 kDa, about 50 kDa, about 60 kDa, about 70 kDa, about 80 kDa, about 100 kDa, about 200 kDa, about 300 kDa, about 400 kDa, about 500 kDa, about 750 kDa, or about 900 kDa or more and / or about 1000 kDa, about 750 kDa, about 500 kDa, about 400 kDa, about 300 kDa, about 200 kDa, or about 100 kDa or less.

33. The method according to any one of claims 1 to 31, wherein the molecular weight of the backbone of the BB polymer is about 90 kDa or 100 kDa.

34. The method according to any one of claims 1 to 32, wherein the molecular weight of the polymeric pendant chain is about 1 kDa, about 5 kDa, about 10 kDa, about 20 kDa, about 30 kDa, about 40 kDa, about 50 kDa, about 60 kDa, about 70 kDa, about 80 kDa, or about 90 kDa or more and / or about 100 kDa, about 90 kDa, about 80 kDa, about 70 kDa, about 60 kDa, about 50 kDa, about 40 kDa, about 30 kDa, about 20 kDa, or about 15 kDa or less.

35. The method according to any one of claims 1 to 33, wherein the molecular weight of the polymeric pendant chain is about 15 kDa or is between about 10 kDa to about 20 kDa.

36. The method according to any one of claims 1 to 34, wherein a grafting ratio of the BB polymer is between about 30 and about 100 %.

37. The method according to any one of claims 1 to 35, wherein a grafting ratio of the BB polymer is about 30%, about 35%, about 40%, about 45%, or about 50% or more and / or about 100%, about 90%, about 80%, about 70%, about 65%, about 60%, about 55%, or about 50% or less.

38. The method according to any one of claims 1 to 36, wherein the grafting ratio of the BB polymer is between about 40 and about 60%, preferably between about 45% and about 55%, most preferably is about 45% or about 55%.

39. The method according to any one of claims 1 to 37, wherein the concentration of the BB polymer in the composition is about 1, about 25, about 50, about 75, about 85, about 90, about 95, about 100, about 150, about 200, about 250, about 300, about 350, about 400, about 450, or about 500 pg / ml or more and / or about 40, 35, 30, 25, 20, 15, 10, about 5, about 1, about 0.5, about 0.25, or about 0.1 mg / mL or less.

40. The method according to any one of claims 1 to 38, wherein the concentration of the BB polymer is about 20 mg / ml to about 40 mg / ml.

41. The method according to any one of claims 1 to 39, wherein the composition is free of linear polymer.

42. The method according to any one of claims 1 to 40, wherein the composition further comprises a linear polymer.

43. The method according to claim 41, wherein the linear polymer is dextran, poly(vinylpyrrolidone), polyethylene glycol), hydroxypropyl cellulose, a polymethacrylate polymer or copolymer, a polyoxazoline polymer, or a polyacrylate polymer or copolymer, or a (preferably pharmaceutically acceptable) salt thereof.

44. The method according to claim 42, wherein the linear polymer is poly(vinylpyrrolidone), or a (preferably pharmaceutically acceptable) salt thereof.

45. The method according any one of claims 41 to 43, wherein the linear polymer is partially crosslinked.

46. The method according to any one of claims 41 to 44, wherein the linear polymer has a molecular weight of about 5 kDa, about 10 kDa, about 25 kDa, about 50 kDa, about 100 kDa, about 250 kDa, or about 500 kDa or more and / or 10 MDa, about 8 MDa, or about 5 MDa or less.

47. The method according to any one of claims 41 to 45, comprising the BB polymer and the linear polymer in a BB polymer: linear polymer weight ratio between about 1:1 and about 1:20,preferably between about 1:2.5 and about 1:15, more preferably between about 1:5 and about 1:10.

48. The method according to any one of claims 1 to 46, wherein the composition further comprises a pharmaceutically acceptable carrier or excipient.

49. The method according to any one of claims 1 to 46, wherein the pharmaceutically acceptable carrier or excipient comprises a saline solution, preferably isotonic, preferably buffered at a pH of about 7 to about 7.4.

50. The method according to any one of claims 1 to 46, wherein the pharmaceutically acceptable carrier or excipient comprises phosphate-buffered saline (PBS).

51. The method according to any one of claims 47 to 49, wherein the pharmaceutically acceptable carrier or excipient comprises one or more additives such as preservatives, stabilizers, antioxidants, surfactants, viscosity modifiers, and / or chelating agents.

52. The method according to any one of claims 1 to 50, wherein the composition further comprises one or more additional therapeutic agents, or wherein the method further comprises administering one or more additional therapeutic agents.

53. The method according to claim 51, wherein the one or more additional therapeutic agents comprise an analgesic, an anti-inflammatory agent, an antioxidant or a biologic.

54. The method according to claim 52, wherein the anti-inflammatory agent comprises a nonsteroidal anti-inflammatory drug (NSAID) ora glucocorticoid.

55. The method according to any one of claims 1 to 53, wherein the joint disease or condition is joint injury or arthritis.

56. The method according to claim 54, wherein the joint disease or condition is osteoarthritis (OA).

57. The method according to any one of claims 1 to 55, wherein the joint damage and / or pain is knee joint damage and / or pain.

58. The method according to any one of claims 1 to 56, wherein the composition is injected in the joint or in the vicinity of the joint.

59. The method according to any one of claims 1 to 57, wherein the subject is a human.

60. The method according to any one of claims 1 to 58, wherein the subject is a companion animal, such as a dog or a cat, or a farm animal such as an ovine, caprine or bovine, or a sport animal such as a horse.

61. A composition as defined in any one of claims 1 to 54, for use in preventing or reducing joint damage and / or pain in a subject suffering from a joint disease or condition.

62. The composition for use according to claim 60, wherein the joint disease or condition is joint injury or arthritis.

63. The composition for use according to claim 61, wherein the joint disease or condition is osteoarthritis.

64. The composition for use according to any one of claims 60 to 62, wherein the joint damage and / or pain is knee joint damage and / or pain.

65. The composition for use according to any one of claims 60 to 63, wherein the composition is for injection in the joint or in the vicinity of the joint.

66. The composition for use according to any one of claims 60 to 64, wherein the subject is a human.

67. The composition for use according to any one of claims 60 to 64, wherein the subject is a companion animal, such as a dog or a cat, or a farm animal such as an ovine, caprine or bovine, or a sport animal such as a horse.

68. Use of a composition as defined in any one of claims 1 to 53, for preventing or reducing joint damage and / or pain in a subject suffering from a joint disease or condition.

69. Use of a composition as defined in any one of claims 1 to 53, for the manufacture of a medicament for preventing or reducing joint damage and / or pain in a subject suffering from a joint disease or condition.

70. The use according to claim 67 or 68, wherein the joint disease or condition is joint injury or arthritis.

71. The use according to any one of claims 67 to 69, wherein the joint disease or condition is osteoarthritis.

72. The use according to any one of claims 67 to 70, wherein the joint damage and / or pain is knee joint damage and / or pain.

73. The use according to any one of claims 67 to 71, wherein the composition is for injection in the joint or in the vicinity of the joint.

74. The use according to any one of claims 67 to 72, wherein the subject is a human.

75. The use according to any one of claims 67 to 72, wherein the subject is a companion animal, such as a dog or a cat, or a farm animal such as an ovine, caprine or bovine, or a sport animal such as a horse.

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