Cyclodextrin complexes for specifying

Abstract

The present invention provides complexes of a specialized proregression mediator (SPM), or a salt, ester or amide thereof, with cyclodextrin (CD) wherein the SPM is a hydroxylated polyunsaturated fatty acid derived from arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid or docosapentaenoic acid, having a length of 20 or 22 carbon atoms, having a length of 20 or 22 carbon atoms, having a length of 20 or 22 carbon atoms, having a length of 20 or 22 carbon atoms, and having a length of 20 or 22 carbon atoms; 4, 5 or 6 conjugated double bonds are disposed in a diene, triene or tetraene system or a combination thereof; and related compositions and methods.

Description

Technical Field

[0001] The present invention relates to inclusion complexes of cyclodextrins and specialized pro-resolving mediators (referred to herein as "SPMs") including resolvins, protectins, lipoxins and maresins, and their use in therapy. Background Art

[0002] There are two phases of inflammatory response in humans: initiation and resolution. At the cellular level, initiation is a protective response characterized by the production and release of proinflammatory mediators to activate the innate immune response, leading to an influx of polymorphonuclear cells (PMNs) from the blood to kill and clear foreign invaders and damaged cells. This initiation phase is an active process driven by arachidonic acid metabolites including prostaglandins, which are chemoattractants for eosinophils, neutrophils, and monocytes, and leukotrienes, which cause adhesion, chemotaxis, and aggregation of leukocytes.

[0003] During the peak of inflammation, with the elimination of inflammatory stimuli, the resolution phase begins, which is marked by the reduction of proinflammatory signals and the generation and release of specialized pro-resolution mediators (SPMs), which include resolvins, protectins, lipoxins and macrophage mediators. SPM is a hydroxylated polyunsaturated fatty acid derived from arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid or docosapentaenoic acid, with a length of 20 or 22 carbon atoms, and 4, 5 or 6 double bonds are arranged in diene, triene and / or tetraene systems. As an endogenous pro-resolution counterpart of prostaglandins and leukotrienes, SPM drives the resolution phase by preventing PMN infiltration and stimulating the recruitment and activation of monocytes and pro-resolution macrophages, thereby clearing cell debris in tissues in a non-inflammatory manner, restoring tissues to homeostasis (Serhan CN et al., Cold Spring Harb Perspect Biol 2015; 7: a016311).

[0004] Unresolved inflammation is an important but underestimated driver of many chronic diseases and conditions. Therefore, the ability to resolve excessive inflammation is critical to human health (Serhan CN, Nature 2014 510:92-101; Coussens et al., Nature 2002; 420, 860–867; Grivennikov et al., Cell. 2010; 140(6):883-99; Todoric et al., Cancer Prev Res. 2016; 9(12):895-905; Fishbein et al., Pharmacol Ther. 2021; 218:107670).

[0005] In vitro studies of SPM have demonstrated effective effects on inflammation and other markers at nanomolar concentrations. In vivo studies of SPM in a wide range of inflammatory disease and tumor models have demonstrated effects on disease progression and symptoms. These data provide a scientific basis for the therapeutic administration of SPM to treat chronic inflammatory diseases and cancer. (Fullerton JN, Gilroy DW., Nat Rev Drug Discov. 2016; 15(8): 551-567; Dalli J, Serhan CN., Br J Pharmacol. 2019; 176(8): 1024-1037; Sulciner ML, Serhan CN, Gilligan MM et al., J Exp Med. 2018; 215(1): 115-14; Gilligan MM, Gartung A, Sulciner ML, Proc Natl Acad Sci US A. 2019; 116(13): 6292-6297).

[0006] To translate these unique and promising in vitro and in vivo findings into clinical benefit, there is a need to develop compositions capable of delivering SPMs, including their analogs and derivatives, to target tissues in therapeutically effective amounts. The present invention addresses this need by providing stable SPMs complexed with cyclodextrins. Summary of the invention

[0007] The present invention relates to complexes of SPM and cyclodextrin (referred to herein as "SPM complexes") for treating inflammatory diseases and cancer. The SPM complexes and methods described herein are based on the unexpected discovery that the chemical and physical stability of the SPM portion is improved when complexed with cyclodextrin and exposed to air at elevated temperatures and humidity, thereby improving the ability to prepare, formulate, package, stock, distribute and utilize SPM pharmaceutical compositions to prevent and treat inflammatory diseases and cancer. More specifically, the present invention relates to SPM complexes and their use in pharmaceutical compositions, wherein the stability of the SPM active portion is significantly improved.

[0008] The present invention provides a complex of a specialized pro-resolving mediator (SPM) or a salt, ester or amide thereof with a cyclodextrin (CD), wherein the SPM is a hydroxylated polyunsaturated fatty acid derived from arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid or docosapentaenoic acid, having a length of 20 or 22 carbon atoms and having 4, 5 or 6 conjugated double bonds placed in a diene, triene or tetraene system or a combination thereof.

[0009] In embodiments, the SPM is selected from the group consisting of RvE1, RvE2, RvE3, RvE4, AT-RvE1, AT-RvE2, AT-RvE3, RvD1, RvD2, RvD3, RvD4, RvD5, RvD6, AT-RvD1, AT-RvD2, AT-RvD3, AT-RvD4, AT-RvD5, AT-RvD6, RvT1, RvT2, RvT3, RvT4, LXA 4. The group consisting of LXB4, AT-LXA4, AT-LXB4, PD1, PDX, AT-PD1, MaR1 and MaR2, PD1n-3DPA or MaR1n-3DPA or their salts, esters or amides; optionally, wherein the SPM is RvE1, AT-RvE1, RvD1, AT-RvD1, RvD2, AT-RvD1, AT-RvD2, LxA4 or AT-LxA4 or their salts, esters or amides.

[0010] In an embodiment, the cyclodextrin is selected from α-cyclodextrin (α-CD), β-cyclodextrin (β-CD), γ-cyclodextrin (γ-CD), 2-hydroxypropyl-β-cyclodextrin (HP-β-CD), sulfobutyl ether β-cyclodextrin (SBE-β-CD); 2-hydroxypropyl-α-cyclodextrin; randomly methylated β-cyclodextrin; 2-O-methyl-β-cyclodextrin; 2,6-di-O-methyl-β-cyclodextrin; heptathiol (2,3,6-tri-O-methyl)-β-cyclodextrin; carboxymethyl-β-cyclodextrin; carboxyethyl-β-cyclodextrin; hydroxyethyl-β-cyclodextrin; maltosyl-β-cyclodextrin; 3,6-(N,N,N-trimethylammonium)propyl-β-cyclodextrin; acetyl-β-cyclodextrin; 2,6-di-O-methyl-γ-cyclodextrin; 2-hydroxypropyl-γ-cyclodextrin; or sulfobutyl ether γ-cyclodextrin; optionally, wherein the cyclodextrin is α-cyclodextrin (α-CD), β-cyclodextrin (β-CD), γ-cyclodextrin (γ-CD), 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutyl ether β-cyclodextrin (SBE-β-CD); optionally, wherein the cyclodextrin is γ-cyclodextrin (γ-CD) or 2-hydroxypropyl-β-cyclodextrin (HP-β-CD).

[0011] In embodiments, the SPM is present in an amount of weight percentage relative to the total weight of the complex (including only the weight of the SPM and cyclodextrin molecules) ranging from 1.5% to 35%, 2% to 30%, 3% to 25% or 4% to 20%.

[0012] In embodiments, the SPM and cyclodextrin are present in a molar ratio ranging from 2:1 to 1:4, or optionally 1:1 to 1:3, or 1:1.5 to 1:2.5, or 1:2.

[0013] In embodiments, the SPM is in the form of a salt. In embodiments, the SPM is in the form of a sodium salt, a potassium salt, a calcium salt, a zinc salt or a magnesium salt. In embodiments, the SPM is a magnesium salt of dilysine. In embodiments, the SPM is a sodium salt. In embodiments, the SPM is a sodium salt of RvE1.

[0014] In an embodiment, the SPM is in the form of a salt described by Formula I:

[0015]

[0016] in

[0017] M is a divalent metal selected from magnesium (Mg2+), calcium (Ca2+) and zinc (Zn2+);

[0018] A1 and A2 are each the same SPM molecule;

[0019] R1 and R2 are each independently a C1-C10 alkyl group containing at least one basic functional group; and X1 and X2 are each independently H or CO-Z, and Z is a peptide containing 1 to 5 amino acids.

[0020] In an embodiment of Formula I, M is selected from magnesium (Mg2+) or calcium (Ca2+).

[0021] In an embodiment of Formula I, R1 and R2 are each independently -(CH2)3-Y1 and -(CH2)4-Y2, and Y1 and Y2 are each selected from a positively charged primary amine, a positively charged secondary amine, a positively charged tertiary amine and a positively charged guanidine.

[0022] In an embodiment of Formula I, X1 and X2 are each H.

[0023] In an embodiment of Formula I, R1 and R2 are each -(CH2)4-Y2, and Y2 is -NH3+.

[0024] In an embodiment of Formula I, M is magnesium (Mg2+), R1 and R2 are each -(CH2)4-Y2, Y2 is -NH3+, X1 and X2 are each H, A1 and A2 are RvE1, and the SPM of the complex is referred to as RvE1 magnesium dilysate, or "RvE1 MgLys".

[0025] In an embodiment of Formula I, M is magnesium (Mg2+), R1 and R2 are each -(CH2)4-Y2, Y2 is -NH3+, X1 and X2 are each H, A1 and A2 are RvD1, and the SPM of the complex is referred to as RvD1 magnesium dilysate, or "RvD1 MgLys".

[0026] In an embodiment of Formula I, M is magnesium (Mg2+), R1 and R2 are each -(CH2)4-Y2, Y2 is -NH3+, X1 and X2 are each H, A1 and A2 are RvD2, and the SPM of this complex is referred to as RvD2 magnesium dilysate, or "RvD2 MgLys".

[0027] In an embodiment of Formula I, M is magnesium (Mg2+), R1 and R2 are each -(CH2)4-Y2, Y2 is -NH3+, X1 and X2 are each H, A1 and A2 are LxA4, and the SPM of this complex is referred to as LxA4 magnesium dilysate, or "LxA4 MgLys".

[0028] In embodiments of the complex comprising the SPM of Formula I, the cyclodextrin component of the complex is γ-cyclodextrin (γ-CD) or 2-hydroxypropyl-β-cyclodextrin (HP-β-CD).

[0029] The present invention also provides a method for preparing the complex described herein. In an embodiment, the method comprises combining a certain amount of SPM or a salt thereof with a certain amount of cyclodextrin to form a mixture; dissolving the mixture in water; and isolating the complex by a method comprising one or more filtration to obtain a filtrate, followed by freeze drying of the filtrate, crystallization, co-evaporation or spray drying. In an embodiment, the dissolving step is carried out under nitrogen, for example by stirring under nitrogen for 1-3 hours. A complex prepared by the aforementioned method is also provided.

[0030] In embodiments, the SPM component of the composites described herein has a chemical purity of at least 88% as determined by high pressure liquid chromatography (HPLC) exposed to air at 40°C and 75% relative humidity for at least 8 weeks, or 12 weeks, or 16 weeks.

[0031] In embodiments, the SPM component of the complexes described herein has a chemical purity of at least 90%, or at least 92%, or at least 94%, as determined by HPLC.

[0032] In embodiments, the number of degradation peaks of the SPM component of the composite present at greater than or equal to 1.0% as determined by HPLC is less than 8, less than 6, or less than 4, or no more than 2. In embodiments, the number of degradation peaks of the SPM component of the composite present at greater than or equal to 0.2% as determined by HPLC is less than 16, optionally less than 14 or less than 12 or less than 10, or no more than 8.

[0033] The present invention also provides a complex prepared by the method described herein.

[0034] The present invention also provides a composition comprising the complex described herein, and one or more pharmaceutically acceptable carriers and / or excipients.

[0035] In an embodiment, the composition is formulated as an oral, pulmonary, rectal, parenteral, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, inhalation, buccal, sublingual, intrapleural, intrathecal, or intranasal dosage form; optionally, wherein the composition is formulated as an oral or parenteral dosage form.

[0036] In an embodiment, a composition as described herein is used in a method of treating a gastrointestinal inflammatory disease or disorder selected from inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, proctitis, pouchitis, Crohn's disease of the pouch, eosinophilic colitis, lymphocytic colitis, collagenous colitis, diversion colitis, chemical colitis, ischemic colitis, eosinophilic esophagitis, Behcet's disease, irritable bowel syndrome, Celiac disease, intestinal mucositis, diverticulitis, and short bowel syndrome, optionally wherein the inflammatory disease or disorder is ulcerative colitis, Crohn's disease, or pouchitis.

[0037] In embodiments, the compositions described herein are used in a method of treating cancer, optionally wherein the cancer is a solid tumor cancer, such as brain cancer, breast cancer, bladder cancer, colorectal cancer, esophageal cancer, gastric cancer, head and neck cancer, liver cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, or sarcoma, optionally wherein the cancer is colorectal cancer, gastric cancer, liver cancer, lung cancer, ovarian cancer, or pancreatic cancer.

[0038] In an embodiment, the composition is administered orally or parenterally, optionally wherein the parenteral administration is subcutaneous, intraperitoneal, intramuscular or intravenous; optionally wherein the pharmaceutical composition is administered sublingually or by inhalation. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 :Structure and atom number assignment of RvE1-MgLys.

[0040] Figure 2A -C: Structures and atomic number assignments of α-cyclodextrin (α-CD, A), β-cyclodextrin (β-CD, B), and γ-cyclodextrin (γ-CD, C).

[0041] Figure 3A -B: Structures and atomic number assignments of 2-hydroxypropyl-β-cyclodextrin (HP-β-CD, A) and sulfobutyl ether β-cyclodextrin (SBE-β-CD, B).

[0042] Figure 4 : 1H NMR spectrum with full signal assignment of RvE1-MgLys not complexed with cyclodextrin.

[0043] Figure 5 : 2D ROESY spectrum of RvE1-MgLys not complexed with cyclodextrin.

[0044] Figure 6 : 1H NMR spectrum of RvE1-MgLys complexed with α-CD with full signal assignment.

[0045] Fig. 7A -F: Stacked partial 1H NMR spectra of the unsaturated region of RvE1-MgLys alone (A, first from the bottom) and RvE1-MgLys in complex with α-CD, β-CD, γ-CD, HP-β-CD, and SBE-β-CD (BF, respectively).

[0046] Fig. 8A -F: Stacked partial 1H NMR spectra of the saturation region of RvE1-MgLys alone and RvE1-MgLys in complex with α-CD, β-CD, γ-CD, HP-β-CD and SBE-β-CD, AF from the bottom, respectively.

[0047] Fig. 9 : 2D ROESY spectrum of RvE1-MgLys complexed with α-CD with full signal assignment.

[0048] Fig.10 : Partial 2D ROESY spectrum of RvE1-MgLys complexed with α-CD.

[0049] Fig.11 : 1H NMR spectrum of RvE1-MgLys complexed with β-CD with integration and full signal assignment.

[0050] Fig.12 : 2D ROESY spectrum of RvE1-MgLys complexed with β-CD with full signal assignment.

[0051] Fig.13 : Magnified portion 2D ROESY spectrum of RvE1-MgLys complexed with β-CD.

[0052] Fig.14 : 1H NMR spectrum of RvE1-MgLys complexed with γ-CD with integration and full signal assignment.

[0053] Fig.15 : 2D ROESY spectrum of RvE1-MgLys complexed with γ-CD with full signal assignment.

[0054] Fig.16 : Magnified portion 2D ROESY spectrum of RvE1-MgLys complexed with γ-CD.

[0055] Fig.17: 1H NMR spectrum of RvE1-MgLys complexed with HP-β-CD with integration and full signal assignment.

[0056] Fig.18 : 2D ROESY spectrum of RvE1-MgLys complexed with HP-β-CD with full signal assignment.

[0057] Fig.19 : Magnified 2D ROESY spectrum of RvE1-MgLys complexed with HP-β-CD.

[0058] Fig. 20 : 1H NMR spectrum of RvE1-MgLys complexed with SBE-β-CD with integration and full signal assignment.

[0059] Fig.21 : 2D ROESY spectrum of RvE1-MgLys complexed with SBE-β-CD with full signal assignment.

[0060] Fig. 22 : Magnified portion 2D ROESY spectrum of RvE1-MgLys complexed with SBE-β-CD.

[0061] Fig.23A -E: Schematic diagram illustrating the molecular orientation of RvE1 in the cavities of α-CD, β-CD, γ-CD, HP-β-CD, and SBE-β-CD, AE, respectively.

[0062] Fig.24A -D: Photographs of RvE1MgLys alone and RvE1MgLys complexed with γ-CD samples exposed to air under ambient conditions at baseline, 8 hours, 24 hours, and 168 hours.

[0063] Fig.25A - B: Photographs of the direct mixture of RvE1 MgLys complexed with γ-CD and standard pharmaceutical excipients without additional granulation before tableting (A) and after compression into core tablets (B).

[0064] Fig.26 : Schematic representation of the complex of RvE1 magnesium dilysinate (RvE1 MgLys) and γ-cyclodextrin (γ-CD). DETAILED DESCRIPTION

[0065] An object of the present invention is to provide SPM complexes, their preparation and their use in pharmaceutical compositions, wherein the SPM active part is stable to chemical degradation.

[0066] Despite their promising pharmacological effects, SPMs are chemically unstable, in part because their conjugated double bond systems tend to isomerize into thermodynamically more favorable states, a tendency first discovered in aqueous environments such as phosphate buffer solutions and tissue culture media (Maddipati et al., Prostaglandins Other Lipid Mediat. 2011; 94(1-2): 59-72). SPMs are hydroxylated polyunsaturated fatty acids derived from eicosapentaenoic acid, docosahexaenoic acid, or docosapentaenoic acid, 20 or 22 carbon atoms in length, with 5 or 6 double bonds in the diene, triene, and / or tetraene systems. For example, the 6Z, 8E, 10E triene system of resolvin E1 (RvE1) is thermodynamically unstable, resulting in its isomerization into a lower energy all-trans structure. This isomerization results in conversion of the 6,7-cis double bond to the 6,7-trans isomer which has significantly reduced biological activity, showing a 70% reduction in activity in various assays (Lombardo Thesis, Curtin University, January 2015, pp. 33-34).

[0067] In view of the inherent fragility of the conjugated double bond structure of SPM, especially the tendency of diene, triene and tetraene systems to isomerize to the weaker trans configuration, structural modifications have been proposed to stabilize SPM. Lombardo proposed structural modifications of RvE1 to stabilize the molecule, especially by forming benzene analogs to block the isomerization of the 6,7-cis double bond. It has been proposed to use cyclopropane homologues and benzene homologues of resolvin E2 (RvE2) to stabilize its double bond structure (Fukuda H et al., Org Lett. 2016; 18(24): 6224-6227; Murakami Y et al., ACS Med Chem Lett. 2020; 11(4): 479-484). A similar strategy was proposed for lipoxin A4 (LxA4), by inserting a benzo-fused ring system to counteract the tendency of the 11Z double bond in the 7E, 9E, 11Z, 13E tetraene system of LxA4 to isomerize to the all-trans configuration (Petasis et al., Bioorg Med ChemLett. 2008; 18(4): 1382-138). Similarly, the use of a benzo-fused ring was proposed to stabilize resolvin D1 (RvD1) against isomerization of its tetraene system (Orr et al., Am J Physiol Lung Cell Mol Physiol. 2015; 308(9): L904-L911). Although these structural modifications can stabilize SPM against isomerization, they can also result in reduced potency compared to the native, unmodified SPM molecule. For example, Lombardo reported that benzo-lipoxin E1 analogs had fourfold lower binding affinity for a known RvE1 receptor compared to the native RvE1 molecule (Lombardo 2015, p. 113). Similarly, the benzo-lipoxin A4 compounds described by Petasis were less bioactive and 20% to 50% less potent than control lipoxin A4 analogs (Table 2, Petasis 2008). In addition to potential loss of bioactivity, structural SPM analogs may present additional safety and / or efficacy issues due to off-target effects compared to the native, unmodified SPM molecule.

[0068] An alternative strategy is described in WO2017210604, which demonstrates that the formation of mineral amino acid chelate salts of natural SPMs, such as magnesium resolvin E1 dilysinate (RvE1-MgLys), increases their solid-state chemical stability under standard conditions, i.e., 20-24°C, 40% relative humidity, when exposed to air. However, further studies by the inventors have shown that these compounds exhibit undesirable levels of chemical instability over time when exposed to air under accelerated conditions of 40°C, 75% relative humidity. The present invention addresses the need to further stabilize SPMs against chemical instability when exposed to air under elevated temperature and humidity conditions, which reduces the risk of loss of potency and / or the appearance of unknown impurities with potential safety issues during preparation, formulation, packaging, storage and distribution.

[0069] The present invention is based in part on the unexpected discovery that complexing with cyclodextrins stabilizes the fragile triene structure of RvE1. Without being bound by any theory, it is believed that the unique orientation of RvE1 within various cyclodextrins protects the triene system against degradation caused by exposure to light, air and humidity. For other SPMs with similar chemical structures, similar anti-isomerization stability is expected, especially conjugated double bond systems composed of dienes, trienes and / or tetraene systems, which include the SPMs described in Table 1 below.

[0070] Thus, the present invention provides complexes of SPM or its salts or esters or amides of SPM molecules with cyclodextrins, wherein the SPM comprises a diene, triene and / or tetraene system, as well as methods for preparing the complexes and methods of use thereof in therapy. These complexes are advantageously stabilized to resist chemical degradation, particularly when exposed to air under conditions of elevated temperature and humidity (i.e., 40° C. and 75% relative humidity) for a period of up to 16 or 24 weeks, which would otherwise result in significant degradation of the SPM molecule and an increase in degradation products and a concomitant decrease in chemical purity, which together would hinder the use of SPM as a drug. For example, after 16 weeks of exposure to air under the above-mentioned elevated temperature and humidity conditions, the RvE1 portion of RvE1-MgLys alone was significantly degraded from 94.0% to 80.7%, while the RvE1 portion of five different cyclodextrins complexed with RvE1-MgLys was degraded from 95.4% to 91.7% on average. Thus, cyclodextrins reduced RvE1 degradation by an average of 70% and γ-cyclodextrin reduced RvE1 degradation by 91% compared to RvE1-MgLys alone. As used herein, the term "chemical purity" refers to the amount of the active portion of a particular compound in a sample of a compound, given as an area percentage. Unless otherwise indicated, the percentages stated throughout the specification with respect to chemical purity are area percentages, calculated as the ratio of (a) the peak area corresponding to the SPM active portion, isomers, diastereomers, other impurities or degradation products to (b) the sum of the peak areas of the detectable components of the sample as measured by high pressure liquid chromatography (HPLC).

[0071] Although cyclodextrins are known to affect the chemical and photostability of various small molecules, including eicosanoids, a comprehensive review by Popielec and Loftsson in 2017 also identified several molecules that were destabilized by cyclodextrins, highlighting the fact that the effect of a particular cyclodextrin on the stability of any particular molecule is unpredictable (Popielec A, Loftsson T., Int J Pharm. 2017; 531(2): 532-542). For example, a study of various cyclodextrins complexed with prostaglandin E1 analogs in solution (phosphate buffer, pH 8.0, 60°C) showed that six of the eight tested cyclodextrins accelerated the degradation of the analogs compared to controls (see Table 1, Uekama K et al., Pharm Res. 2001; 18(11): 1578-1585). Furthermore, molecules stabilized by cyclodextrins in aqueous solutions can be destabilized by the same cyclodextrins in solid dosage forms (Hamada Y et al., Chem Pharm Bull (Tokyo). 1975; 23(6): 1205-1211; Uekama 2006; Popielec 2017). According to Uekama 2006, "cyclodextrins are known to accelerate and decelerate various reactions depending on the nature of the complex formed". In particular, the degree of inclusion and orientation of the guest molecule in the cyclodextrin, especially relative to the reactive site of the molecule, plays a key role in determining the stabilizing effect of the cyclodextrin (Popielec 2017). Therefore, "the use of cyclodextrins to improve the stability of a given drug is unpredictable and must be thoroughly studied and carefully formulated" (Rincón-López 2021).

[0072] Compound

[0073] The present invention relates to a complex of SPM molecules or their salts or esters or amides and cyclodextrin.

[0074] Cyclodextrin component of the complex

[0075] The term "cyclodextrin" refers to a cyclic oligosaccharide composed of at least six pyranose glucose units connected by α-1,4 glycosidic bonds, wherein the oligosaccharide ring forms a ring or cone-shaped structure. In the context of the present invention, cyclodextrins with six glucose subunits are called α-cyclodextrins, cyclodextrins with seven glucose subunits are called β-cyclodextrins, and cyclodextrins with eight glucose subunits are called γ-cyclodextrins. These cyclodextrins are collectively referred to as natural cyclodextrins. In addition, the term "cyclodextrin" also refers to randomly substituted derivatives of natural cyclodextrins, including but not limited to 2-hydroxypropyl-α-cyclodextrin; randomly methylated β-cyclodextrin; 2-O-methyl-β-cyclodextrin; 2,6-di-O-methyl-β-cyclodextrin; 2-hydroxypropyl-β-cyclodextrin, heptath(2,3,6-tri-O-methyl)-β-cyclodextrin; carboxymethyl-β-cyclodextrin; carboxyethyl-β-cyclodextrin; hydroxyethyl-β-cyclodextrin; maltosyl-β-cyclodextrin; 3,6-(N,N,N-trimethylammonium)propyl-β-cyclodextrin; acetyl-β-cyclodextrin; sulfobutyl ether β-cyclodextrin; 2,6-di-O-methyl-γ-cyclodextrin; 2-hydroxypropyl-γ-cyclodextrin; and sulfobutyl ether γ-cyclodextrin.

[0076] The term "complex" refers to an inclusion compound in which a "guest" molecule is partially or completely contained in the void space or cavity of a "host" molecule, or in the crystal lattice of a host molecule. In the context of the present invention, a "complex" is an inclusion compound formed by an SPM "guest" molecule and a cyclodextrin "host" molecule (also referred to herein as an "SPM complex").

[0077] SPM components of the complex

[0078] The present invention relates to a complex of a specialized pro-resolving mediator (SPM) or a salt thereof or an ester or amide thereof with a cyclodextrin, wherein the SPM comprises a diene, a triene and / or a tetraene system. SPMs include resolvins, protectins, lipoxins and macrophage mediators and their aspirin-triggered counterparts (e.g., aspirin-triggered lipoxins and protectins). These molecules are described in, for example, US 5,441,951 and US 8,119,691 (lipoxins and aspirin-triggered lipoxins), US 6,670,396 (aspirin-triggered lipid mediators), US2006-0293288 (resolvins), US 7,378,444 and US 7,595,341 (lipid mediator analogs derived from ω-3 fatty acids). The esters of SPM can be isopropyl esters, methyl esters, ethyl esters or glycerol esters (Obrosov A et al., J Neurol Neurophysiol. 2017; 8(6)).

[0079] In embodiments, the SPM component of the complex described herein is selected from the molecules listed in Table 1. In embodiments, the SPM component of the complex described herein is a free acid of a SPM described in Table 1 or a salt, or ester, or amide thereof. In an embodiment, the SPM component of the complex described herein is selected from RvE1, RvE2, RvE3, RvE4, AT-RvE1, AT-RvE2, AT-RvE3, RvD1, RvD2, RvD3, RvD4, RvD5, RvD6, AT-RvD1, AT-RvD2, AT-RvD3, AT-RvD4, AT-RvD5, AT-RvD6, RvT1, RvT2, RvT3, RvT4, LXA4, LXB4, AT-LXA4, AT-LXB4, PD1, PDX, AT-PD1, MaR1 and MaR2, PD1n-3DPA or MaR1n-3DPA or a salt, ester or amide thereof.

[0080] Table 1: SPM components of the composite

[0081]

[0082]

[0083]

[0084]

[0085] *DPA n-3 refers to SPM derived from docosapentaenoic acid, in which the terminal carbon-carbon double bond is located at the ω position.

[0086] Salt

[0087] The SPM component of the SPM-cyclodextrin complex described herein can be a salt of the SPM molecule. In an embodiment, the salt of the SPM molecule can be a pharmaceutically acceptable base addition salt, including a salt formed with a basic organic or inorganic part; or a salt described by formula I, or a dipeptide and polypeptide salt described in WO2017210604 or WO2019108605.

[0088] Base addition salt

[0089] In an embodiment, an SPM salt consisting of a pharmaceutically acceptable base addition salt is prepared by adding an inorganic base or an organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Preferred organic salts are derived from organic bases. They include, but are not limited to, the following salts: primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and alkaline ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, meglumine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, and the like. Particularly preferred organic bases are lysine, isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.

[0090] In an embodiment, the SPM salt is described by formula I and comprises two molecules of the same SPM, which are ionically bonded to two basic functional groups provided by the scaffold described in formula I, wherein the scaffold is a divalent metal-amino acid chelate or a divalent metal-peptide chelate. Typically, the carboxylic acid portion of the SPM molecule or the molecule forming the SPM component is deprotonated to form an ionic bond with the basic functional group (or multiple functional groups) of the compound scaffold portion. These scaffolds and their SPM salts are described in WO2017210604 and WO2019108605. According to the present invention, the SPM of the SPM component that can form the salt of formula I is given in Table 1. It should be understood that if solvated at an appropriate pH, the neutral compound described in the table can be charged (i.e., deprotonated) and form the anionic component described in the compound of formula I. The compounds represented by formula I each contain two SPM molecules, which can be referred to as the "SPM component" of the compound herein, and the scaffold portion to which the SPM component is ionically bonded. The term "bis" refers to two (two) SPM molecules in a salt compound. In an embodiment, the two SPM molecules are identical and are selected from RvE1, RvE2, RvE3, RvE4, AT-RvE1, AT-RvE2, AT-RvE3, RvD1, RvD2, RvD3, RvD4, RvD5, RvD6, AT-RvD1, AT-RvD2, AT-RvD3, AT-RvD4, AT-RvD5, AT-RvD6, RvT1, RvT2, RvT3, RvT4, LXA4, LXB4, AT-LXA4, AT-LXB4, PD1, PDX, AT-PD1, MaR1 and MaR2, PD1n-3DPA or MaR1n-3DPA.

[0091] Abbreviations used herein have their conventional meanings in the chemical and biological arts. The chemical structures and formulae described herein are constructed according to standard rules of chemical valence known in the chemical arts. For example, where a substituent such as -NH3 is shown to be uncharged, it is understood to carry a formal charge, i.e., NH3+.

[0092] Unless otherwise indicated, the term "alkyl" by itself or as part of another substituent refers to a straight (i.e., unbranched) or branched carbon chain (or carbons), or combinations thereof, which may be fully saturated, monounsaturated, or polyunsaturated, and may include monovalent, divalent, and multivalent groups, having the specified number of carbon atoms (i.e., C1-C10 refers to one to ten carbons).

[0093] The term "basic functional group" refers to a positively charged or protonated primary amine, a positively charged secondary amine, a positively charged tertiary amine, or a positively charged guanidine. In an embodiment, a basic functional group refers to -NH3+, -NHC(NH2+)NH2, -NHR6R7, -NR6R7R8, wherein R6, R7, and R8 are each independently hydrogen, -CN, -COOH, -CONH2, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl; R6 and R7 substituents bonded to the same nitrogen atom may be optionally connected to form an unsubstituted heterocycloalkyl or unsubstituted heteroaryl. In an embodiment, a basic functional group is a hydrogen bond acceptor. In an embodiment, a basic functional group is a positively charged amine.

[0094] It is understood that due to resonance, the charge may be distributed throughout the molecule.The chemical structures and formulae described herein are constructed according to standard rules of chemical valency known in the chemical arts, so one skilled in the art will recognize the equivalence of moieties having resonating structures.

[0095] In an embodiment, "amino acid side chain" or "side chain" as used herein is used according to its ordinary meaning to refer to a functional substituent contained on a naturally occurring amino acid. Naturally occurring amino acids are those encoded by the genetic code (e.g., alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine), as well as those amino acids that are later modified, such as hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. In an embodiment, the side chain of an amino acid is ionized (e.g., it has a formal charge).

[0096] In an embodiment, the side chain is selected from H,

[0097]

[0098] The group composed of.

[0099] In an embodiment, the side chain is H. In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is

[0100] In embodiments, a side chain may be optionally attached to the adjacent nitrogen to form an unsubstituted heterocycloalkyl (eg, pyrrolidinyl).

[0101] In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is In an embodiment, the side chain is

[0102] The side chain of glycine is H. The side chain of arginine is The side chain of histidine is The side chain of lysine is The side chain of aspartic acid is The side chain of glutamic acid is The side chain of serine is The side chain of threonine is The side chain of asparagine is The side chain of glutamine is The side chain of cysteine ​​is The side chain of proline is The side chain of alanine is The side chain of valine is The side chain of isoleucine is The side chain of leucine is The side chain of methionine is The side chain of phenylalanine is The side chain of tyrosine is The side chain of tryptophan is

[0103]

[0104] The term "non-natural amino acid side chain" refers to a functional substituent of a compound having the same basic chemical structure as a naturally occurring amino acid, i.e., an alpha carbon bonded to a hydrogen, carboxyl, amino, and R group, such as homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium, allylamine, 2-aminoisobutyric acid. Non-natural amino acids are naturally occurring or chemically synthesized non-protein amino acids. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as naturally occurring amino acids. Non-limiting examples include exo-cis-3-aminobicyclo[2.2.1] hept-5-ene-2-carboxylic acid hydrochloride, cis-2-aminocycloheptylcarboxylic acid hydrochloride, cis-6-amino-3-cyclohexene-1-carboxylic acid hydrochloride, cis-2-amino-2-methylcyclohexanecarboxylic acid hydrochloride, cis-2-amino-2-methylcyclopentanecarboxylic acid hydrochloride, 2-(Boc-aminomethyl)benzoic acid, 2-(Boc-amino)octanedioic acid, Boc-4,5-dehydro-Leu-OH (dicyclohexylammonium), Boc-4-(Fmoc-amino)-L-phenylalanine, Boc-β-Homopyr-OH, Boc-(2-indanyl)-Gly-OH, 4-Boc-3-morpholineacetic acid, 4-Boc-3-morpholineacetic acid, Boc-pentafluoro-D-phenylalanine , Boc-pentafluoro-L-phenylalanine, Boc-Phe(2-Br)-OH, Boc-Phe(4-Br)-OH, Boc-D-Phe(4-Br)-OH, Boc-D-Phe(3-Cl)-OH, Boc-Phe(4-NH2)-OH, Boc-Phe(3-NO2)-OH, Boc-Phe(3,5-F2)-OH, 2-(4-Boc-piperazinyl)-2-(3,4-dimethoxyphenyl)acetic acid purum, 2-(4-Boc-piperazinyl)-2-(2-fluorophenyl)acetic acid purum, 2-(4-Boc-piperazinyl)-2-(3-fluorophenyl)acetic acid purum, 2-(4- 2-(4-Boc-piperazinyl)-2-(4-fluorophenyl)acetic acid purum, 2-(4-Boc-piperazinyl)-2-(4-methoxyphenyl)acetic acid purum, 2-(4-Boc-piperazinyl)-2-phenylacetic acid purum, 2-(4-Boc-piperazinyl)-2-(3-pyridyl)acetic acid purum, 2-(4-Boc-piperazinyl)-2-[4-(trifluoromethyl)phenyl]acetic acid purum, Boc-β-(2-quinolyl)-Ala-OH, N-Boc-1,2,3,6-tetrahydro-2-pyridinecarboxylic acid, Boc-β-(4-thiazolyl)-Ala-OH, Boc-β-(2-thienyl)-D-Ala-OH, Fmo c-N-(4-Boc-aminobutyl)-Gly-OH, Fmoc-N-(2-Boc-aminoethyl)-Gly-OH, Fmoc-N-(2,4-dimethoxybenzyl)-Gly-OH, Fmoc-(2-indanyl)-Gly-OH, Fmoc-pentafluoro-L-phenylalanine, Fmoc-Pen(Trt)-OH, Fmoc-Phe(2-Br)-OH, Fmoc-Phe(4-Br)-OH, Fmoc-Phe(3,5-F2)-OH, Fmoc-β-(4-thiazolyl)-Ala-OH, Fmoc-β-(2-thienyl)-Ala-OH, 4-(hydroxymethyl)-D-phenylalanine. .

[0105] Compounds of formula I

[0106] In an embodiment, the present disclosure provides a compound of Formula I:

[0107]

[0108] in

[0109] M is a divalent metal;

[0110] A1 and A2 are each the same SPM anion;

[0111] R1 and R2 are each independently a C1-C10 alkyl group containing at least one basic functional group;

[0112] X1 and X2 are each independently H or CO-Z, and Z is a peptide comprising 1 to 5 amino acids or a pharmaceutically acceptable salt thereof.

[0113] The compound of formula I has two amino acid moieties and two SPM molecules coordinated around a divalent metal cation as an amino acid component. In embodiments, the divalent metal cation is Mg2+, Ca2+, Mn2+, Fe2+, Cu2+, Co2+, Ni2+, Mo2+ or Zn2+. In embodiments, the divalent metal cation is Mg2+. In embodiments, the divalent metal cation is Ca2+. In embodiments, the divalent metal cation is Zn2+.

[0114] In an embodiment, the amino acid component includes lysine or arginine or consists of lysine or arginine. In an embodiment, the amino acid component includes lysine or arginine. In an embodiment, the basic functional group of R1 and R2 is selected from primary amines, secondary amines, tertiary amines and guanidine. In an embodiment, the basic functional group refers to -NH3, -NHC(NH2+)NH2, -NHR6R7 or -NR6R7R8, wherein R6, R7, R8 are each independently hydrogen, -CN, -COOH, -CONH2, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl or unsubstituted heteroaryl; R6 and R7 substituents bonded to the same nitrogen atom may be optionally connected to form unsubstituted heterocycloalkyl or unsubstituted heteroaryl. In an embodiment, the basic functional group is a hydrogen bond acceptor. In an embodiment, the basic functional group is a hydrogen bond donor. In an embodiment, the basic functional group is a positively charged amine.

[0115] In an embodiment, R1 and R2 are each a side chain of an amino acid residue having a basic functional group. In an embodiment, R1 and R2 are identical, and the amino acid residue is lysine or arginine.

[0116] In an embodiment, R1 and R2 are independently selected from -(CH2)3-Y1 and -(CH2)4-Y2, wherein Y1 and Y2 are each a basic functional group which may be the same or different. In an embodiment, R1 is -CH2CH2NH3. In an embodiment, R2 is -CH2CH2NH3. In an embodiment, R1 is -CH2CH2CH2CH2NH3. In an embodiment, R2 is -CH2CH2CH2CH2NH3.

[0117] In an embodiment, R1 and R2 are both -(CH2)4-Y2, and Y2 is -NH3+.

[0118] In an embodiment, R1 and R2 are both -(CH2)3-Y1, and Y1 is -NHC(NH2+)NH2.

[0119] In an embodiment, R1 is -(CH2)3-Y1, Y1 is -NHC(NH2+)NH2, Y2 is -(CH2)4-Y2, and Y2 is -NH3+. In an embodiment, R1 is -(CH2)4-Y2, Y2 is -NH3+, R2 is -(CH2)3-Y1, and Y1 is NHC(NH2+)NH2.

[0120] In an embodiment, X1 and X2 are the same and are hydrogen (H). In an embodiment, X1 is hydrogen. In an embodiment, X2 is hydrogen.

[0121] In one embodiment of the compound of Formula I, A1 and A2 are selected from RvE1, RvE2, RvE3, RvE4, AT-RvE1, AT-RvE2, AT-RvE3, RvD1, RvD2, RvD3, RvD4, RvD5, RvD6, AT-RvD1, AT-RvD2, AT-RvD3, AT-RvD4, AT-RvD5, AT-RvD6, RvT1, RvT2, RvT3, RvT4, LxA4, LxB4, AT-LxA4, AT-LxB4, PD1, PDX, AT-PD1, MaR1 and MaR2, PD1n-3DPA or MaR1n-3DPA; M is Mg2+, Ca2+ or Zn2+, R1 and R2 are both -(CH2)4-Y2 and Y2 is NH3+; and X1 and X2 are H. This selection of R1, R2 and Y2 may be referred to herein as a metal "dilysine salt", such as "dilysine magnesium" or "dilysine Mg". In this embodiment, the peptide component consists of a lysine dipeptide.

[0122] In one embodiment of the compound of Formula I, A1 and A2 are the same and are selected from the E series resolvins. In an embodiment, the E series resolvins are selected from RvE1, RvE2, RvE3, RvE4, AT-RvE1, AT-RvE2 and AT-RvE3, M is Mg2+, Ca2+ or Zn2+, R1 and R2 are both -(CH2)4-Y2 and Y2 is NH3+; and X1 and X2 are H. This selection of R1, R2 and Y2 may be referred to herein as a metal "dilysinate", such as "dilysinate magnesium" or "dilysinate Mg".

[0123] In one embodiment of the compound of Formula I, A1 and A2 are the same and are selected from the D series resolvins. In an embodiment, the D series resolvins are selected from RvD1, RvD2, RvD3, RvD4, RvD5, RvD6, AT-RvD1, AT-RvD2, AT-RvD3, AT-RvD4, AT-RvD5, AT-RvD6, M is Mg2+, Ca2+ or Zn2+, R1 and R2 are both -(CH2)4-Y2 and Y2 is NH3+; and X1 and X2 are H. This selection of R1, R2 and Y2 may be referred to herein as a metal "dialysinate", such as "dialysinate magnesium" or "dialysinate Mg".

[0124] In one embodiment of the compound of Formula I, A1 and A2 are the same and are selected from LxA4 or AT-LxA4, M is Mg2+, Ca2+ or Zn2+, R1 and R2 are both -(CH2)4-Y2 and Y2 is NH3+; and X1 and X2 are H. This selection of R1, R2 and Y2 may be referred to herein as a metal "dilysinate", such as "dilysinate magnesium" or "dilysinate Mg".

[0125] Physical properties

[0126] Compared to free SPM and its salt, ester or amide, the complexes described herein and compositions comprising the same have advantageous chemical and physical properties. For example, in an embodiment, the SPM component of the complex described herein can be stabilized as a solid or as a solution in deionized water to resist chemical degradation compared to the corresponding free SPM or SPM salt, ester or amide under accelerated conditions of 40° C., 75% relative humidity and when exposed to air. In an embodiment, the complex is stable to chemical degradation, including oxidative degradation. In an embodiment, the complex is stabilized to resist degradation caused by exposure to air, oxygen and humidity, as evidenced by the relative lack of changes in physical properties such as flow characteristics and / or chemical properties measured, for example, by spectroscopic techniques such as nuclear magnetic resonance (NMR) or high pressure liquid chromatography (HPLC). In an embodiment, the relative lack of chemical degradation at 8 or 16 or 24 weeks demonstrates increased stability compared to SPM in the form of a sodium salt or a mineral amino acid chelate. In embodiments, the SPM component of the complexes described herein is stable to chemical degradation as evidenced by significantly reduced degradation products at 8 or 16 or 24 weeks compared to SPM in its salt or ester form.

[0127] In an embodiment, the complex is a solid, free-flowing material in a physical sense, suitable for formulation into solid dosage forms such as powders, tablets, capsules, films or caplets, and parenteral dosage forms such as solutions, suspensions, emulsions and dry powders for reconstitution. For example, in an embodiment, when exposed to air at a temperature of 18 to 22° C. and a relative humidity of 50% to 75%, the complex maintains its form as a solid, free-flowing material for up to 168 hours without visible changes, compared to the corresponding free SPM or SPM salt, ester or amide (which changes from a brown / orange powder to a gel-like or oily substance within 24 hours). In an embodiment, the complex can be mixed with standard pharmaceutical excipients such as fillers, smoothing agents, disintegrants and lubricants, and its flow properties are suitable for gravity feeding in a rotary press. For example, in embodiments, when the composite is mixed with a standard mixture of pharmaceutical excipients in a ratio of about 1 to 4.3 (weight / weight) without further dry or wet grinding (i.e., granulation), the particle size distribution of the resulting mixture has flow characteristics suitable for compression by a gravity-fed rotary press. In embodiments, using the above 1:4.3 mixture, core tablets with a tablet hardness between 18 kilograms force (kilopond) and 30 kilograms force produced using a gravity-fed rotary press have friability and disintegration that meet the specifications considered suitable for an immediate release oral dosage form or suitable for an enteric coating as a sustained release oral dosage form. In addition, the composites and compositions of the present invention can be easily combined.

[0128] Pharmaceutical composition

[0129] The present disclosure provides a pharmaceutical composition comprising a SPM (such as a resolvin, a protectin, a lipoxin or a macrophage mediator) complexed with a cyclodextrin, or a salt or ester thereof, or an amide thereof. In an embodiment, the salt of the SPM can be a simple salt, such as a sodium salt, a potassium salt, a calcium salt, a zinc salt or a magnesium salt, any other pharmaceutically acceptable base addition salt or a salt described by Formula I.

[0130] The pharmaceutical compositions described herein can be formulated for any suitable route of administration, such as oral, pulmonary, rectal, parenteral, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, inhalation, buccal, sublingual, intrapleural, intrathecal, intranasal, vaginal, etc. Dosage forms for topical or transdermal administration of the compounds of the invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants.

[0131] In an embodiment, the pharmaceutical composition is formulated into a parenteral dosage form, such as a sterile aqueous solution or dispersion suitable for parenteral administration. In an embodiment, the parenteral dosage form is selected from an intravenous dosage form, an intraarterial dosage form, or an intramuscular dosage form. In an embodiment, the dosage form is suitable for administration by a subcutaneous route.

[0132] In an embodiment, the pharmaceutical composition is in the form of a sterile aqueous solution or dispersion suitable for administration by direct injection or by addition to a sterile infusion solution for intravenous infusion, and comprises a solvent or dispersion medium containing water, ethanol, a polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), or a suitable mixture thereof.

[0133] According to any of these embodiments, the dosage form can be in the form of a clear aqueous solution, which can be optionally frozen, or in the form of a lyophilized solid, preferably a sterile lyophilized solid, for example contained in a container, such as a prefilled syringe, vial or ampoule. In some embodiments, the container contains a lyophilized composition and is suitable for reconstitution with a specified amount of sterile water, aqueous buffer or other commonly used parenteral diluent (e.g., 5% glucose solution, physiological saline solution, etc.) for administration by a parenteral route, such as intravenous, intraarterial or intramuscular, subcutaneous administration.

[0134] In an embodiment, the pharmaceutical composition is formulated as an oral or peroral dosage form. In an embodiment, the oral formulation is in the form of, for example, a tablet, capsule, powder, solution, suspension or emulsion.

[0135] In an embodiment, the pharmaceutical composition is formulated as a sublingual dosage form. In an embodiment, the sublingual formulation is in the form of a tablet, a film, or a spray.

[0136] In embodiments, the pharmaceutical composition is formulated for administration by nasal or oral inhalation. In embodiments, the inhalable dosage form is a liquid preparation, such as an aqueous solution preparation suitable for pulmonary delivery by a nebulizer, the nebulizer comprising a jet, a vibrating mesh screen and a static mesh screen or a hole nebulizer. In embodiments, the inhalable dosage form is a dry powder for inhalation (DPI). In embodiments, the inhalable dosage form is a propellant-based aerosol preparation suitable for administration using a metered dose inhaler (MDI).

[0137] In an embodiment, the pharmaceutical composition comprising the SPM complex is a unit dosage form of the SPM in the complex. In an embodiment, the unit dosage form is a sterile freeze-dried composition in a suitable container such as an ampoule or a vial. The term "freeze-dried" is synonymous with the term "freeze-dried" herein. In an embodiment, the unit dose comprises 0.5 micrograms (ug) to 100 milligrams (mg) of the SPM active portion. In an embodiment, the unit dose comprises 1, 25, 50, 100, 250 or 500 micrograms of the SPM active portion. In an embodiment, the unit dose comprises 1, 2, 3 or 5 milligrams of the SPM active portion or a multiple thereof, between 1 and 100 milligrams.

[0138] The compositions described herein can be formulated using one or more suitable excipients or carriers. Suitable excipients or carriers are suitable for human or animal use. The term "excipient" refers to an additive that plays a certain role in the composition except for the carrier, for example, as a stabilizer, solubilizer, filler, compression aid, flow modifier, lubricant, disintegrant or suspending agent. Typically, the carrier has a dual purpose, i.e., as a simple carrier or diluent and excipient. Therefore, examples of pharmaceutically acceptable excipients may include carriers. Non-limiting examples of excipients for the compositions of the present invention include sterile liquids, water, buffered saline, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), oils, detergents, suspending agents, carbohydrates (e.g., glucose, lactose, sucrose or dextran), antioxidants (e.g., ascorbic acid or glutathione), chelating agents, low molecular weight peptides and suitable mixtures thereof.

[0139] Suitable excipients or carriers are typically pharmaceutically acceptable carriers or excipients for humans or animals. The term "pharmaceutically acceptable" means approved by a regulatory agency of a federal or state government, or listed in the U.S. Pharmacopoeia or other recognized pharmacopoeias such as the European Pharmacopoeia, for use in animals, more specifically in humans. In the context of the pharmaceutical composition of the present invention, "carrier" refers to, for example, a solvent, diluent or vehicle, with which the complex of the present invention is formulated for delivery. Examples of pharmaceutically acceptable carriers for the compositions of the present invention include, but are not limited to, sterile aqueous and non-aqueous liquids, water, buffered saline, ethanol, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.) and oils for liquid dosage forms; or carbohydrates (such as glucose, lactose, sucrose or dextran) for solid dosage forms.

[0140] The present disclosure also provides packaging and kits containing the pharmaceutical compositions for use in the methods of the present invention. The kit may include one or more containers selected from the group consisting of bottles, vials, ampoules, blister packs, infusion pumps and syringes. The kit may further include one or more instructions for use, one or more syringes, one or more applicators or a sterile solution suitable for reconstitution of the pharmaceutical compositions of the present invention.

[0141] In an embodiment, the pharmaceutical composition is a solid dosage form comprising a SPM complex mixed with one or more excipients or carriers. The mixture of the SPM complex and the excipient can be directly filled into a capsule or compressed into a tablet, and the mixture can be pre-granulated or not. In an embodiment, the SPM complex and the excipient are present in a weight ratio ranging from 10:1 to 1:5, or optionally 5:1 to 1:3, or 3:1 to 1:2, or 2:1 to 1:1.5.

[0142] Method for preparing the composite

[0143] The present invention provides a method for complexing cyclodextrin with SPM or its salt, comprising dissolving SPM or its salt and cyclodextrin in an aqueous solution, which can be, for example, water or an organic solvent / water mixture. According to the method described herein, the pH of the aqueous solution is preferably between 5 and 9. By removing the aqueous solvent, for example by one or more techniques such as crystallization (Loftsson T et al., J Pharm Pharmacol. 2016;68(5):544-555), or by using drying techniques such as reduced pressure co-evaporation, spray drying or freeze drying (Desai NS et al., J. Incl. Phenom. Macrocycl. Chem. 2011;70:217-225; Maestrelli F et al., Eur J Pharm Biopharm. 2011 Aug;78(3):385-93; Jablan J et al., JPharm Biomed Anal. 2012 Dec;71:35-44; Bragagni M. et al., J. Incl. Phenom. Macrocycl. Chem. 2010;68:437-445; Jug M et al., Drug Dev Ind Pharm. 2009 Jul;35(7):796-807; Promzeleva et al., Drug Dev Ind Pharm. 2009 Jul;35(7):810-821; Promzeleva et al., Drug Dev Ind Pharm. 2011 Jul;35(7):810-821; Promzeleva et al., Drug Dev Ind Pharm. 2009 ... M. et al., ACS Biomater. Sci. Eng. 2018; 4:491–501), and the resulting complex was isolated.

[0144] Drug Use

[0145] There are two phases of inflammatory response in humans: initiation and resolution. At the cellular level, initiation is a protective response characterized by the production and release of proinflammatory mediators to activate the innate immune response, leading to an influx of polymorphonuclear cells (PMNs) from the blood to kill and clear foreign invaders and damaged cells. This initiation phase is an active process driven by arachidonic acid metabolites including prostaglandins, which are chemoattractants for eosinophils, neutrophils, and monocytes, and leukotrienes, which cause adhesion, chemotaxis, and aggregation of leukocytes.

[0146] During the peak of inflammation, as the inflammatory stimulus decreases, the resolution phase begins, which is marked by a decrease in proinflammatory signals and a shift from the production and release of proinflammatory mediators to the production and release of SPMs, including resolvins, protectins, lipoxins, and macrophage mediators. SPMs activate and coordinate the resolution phase by preventing PMN infiltration and stimulating the recruitment and activation of monocytes and pro-resolving macrophages, thereby clearing cellular debris from the tissue in a non-inflammatory manner and restoring the tissue to homeostasis (Serhan et al., Cold Spring Harb Perspect Biol 2015; 7: a016311).

[0147] Unresolved inflammation is widely recognized as a unifying aspect of many chronic diseases and disorders. Therefore, the ability to resolve excessive inflammation is critical to human health (Serhan CN, Nature 2014 510:92-101; Coussens et al., Nature 2002; 420, 860–867; Grivennikov et al., Cell. 2010; 140(6):883-99; Todoric et al., Cancer Prev Res. 2016; 9(12):895-905; Fishbein et al., Pharmacol Ther. 2021; 218:107670).

[0148] Researchers have demonstrated the efficacy of SPMs in the prevention and / or treatment of diseases in many preclinical models, including those related to Alzheimer's disease, burns, chronic pancreatitis, diabetic wounds, dermatitis, lung inflammation, peripheral nerve injury, obesity, allergic airway reactions, amyotrophic lateral sclerosis, acute lung injury, fibrosis, bacterial infection, peritonitis, dry eye, tissue regeneration, pain, adipose tissue inflammation, localized aggressive periodontitis, colitis, temporomandibular joint inflammation, arthritis, postoperative pain, postoperative cognitive decline, endotoxin shock, HSV-keratitis, allograft rejection, cardiac ischemia, bacterial pneumonia, cigarette smoke-induced lung inflammation, vascular inflammation, fibromyalgia, and vagotomy (Serhan CN et al., Cold Spring Harb Perspect Biol 2015;7:a016311).

[0149] Lim et al. described the analgesic efficacy of SPM in a large number of inflammatory pain models and characterized resolvins and related substances as therapeutic candidates for preventing inflammatory exacerbations and pathological pain (Lim JY et al., BioMed Research International 2015, pp. 1-14). Lim also pointed out that the "strong efficacy" and "negligible side effects" of these molecules make them attractive candidates for clinical application.

[0150] U.S. Patent Nos. 8,008,282 and 6,627,658 describe lipoxin analogs and their use as angiogenesis inhibitors. U.S. Patent Nos. 5,441,951, 5,648,512, 6,048,897, 6,316,648, 6,569,075, 6,887,901, 7,288,569 and 7,294,728, 7,741,369 and 7,741,369 describe lipoxin compounds and their use in treating cell proliferative disorders. U.S. Patent No. 8,119,691 describes the use of lipoxins and aspirin-triggered lipoxins and their analogs in treating asthma and inflammatory airway diseases.

[0151] US20060293288 describes the use of resolvins for the treatment of gastrointestinal inflammation and diseases such as ulcerative colitis, Crohn's disease, infectious enteritis, antibiotic-associated diarrhea, Clostridium difficile colitis, microscopic or lymphocytic colitis, collagenous colitis, colon polyps, familial polyposis, familial polyposis syndrome, Gardner's Syndrome, Helicobacter pylori, irritable bowel syndrome, nonspecific diarrheal diseases and intestinal cancer. U.S. Patent Nos. 6,670,396, 7,053,230, 7,378,444, 7,585,856, 7,595,341, 7,709,669, 7,737,178, 8,349,896, 8,461,201, 8,569,542 and 8,853,437 describe resolvins and their use in the treatment of inflammation, angiogenesis, cardiovascular disease, thrombosed varicose veins, blood vessel disease, and vascular disease. The invention also provides for use in methods of treating inflammatory bowel diseases, such as Crohn's disease, ulcerative colitis, distal proctitis, rheumatoid spondylitis, arthritis, rheumatoid arthritis, osteoarthritis, gouty arthritis, psoriasis, dermatitis, eczematous dermatitis, atopic or seborrheic dermatitis, allergic or irritant contact dermatitis, fissure eczema, and dermatitis. craquelee), photoallergic dermatitis, phototoxic dermatitis, phytophotosensitive dermatitis, radiation dermatitis, stasis dermatitis, arterial inflammation, coronary artery infarction injury, restenosis, uveitis, iritis, conjunctivitis, adult respiratory distress syndrome, bronchitis, cystic fibrosis, spasmogenic conditions, asthma, idiopathic bronchial asthma, arterial smooth muscle constriction, coronary artery spasm, myocardial infarction, ischemia-induced myocardial damage, cerebral spasm, stroke, inflammatory bowel disease, spastic colon, mucous colitis, allergic conditions, eczema, allergic bowel disease, celiac disease, allergic eye conditions, hay fever, allergic rhinitis, allergic conjunctivitis, conditions involving platelet aggregation, coronary artery thrombosis, phlebitis or venous thrombosis, and methods of treating cardiovascular disease.

[0152] US20120245229 describes resolvins and methods for treating neuropathic pain, including pain associated with diabetic neuropathy or HIV infection, and methods for treating postoperative pain, inflammatory pain, cancer-related pain, and pain associated with fibromyalgia by administering resolvins.

[0153] U.S. Patent No. 10,233,167 describes macrophage mediators and methods for treating or preventing inflammation associated with: neurodegeneration, memory loss, wrinkles, psoriasis, dandruff, dermatitis, arterial inflammation, arthritis, psoriasis, urticaria, vasculitis, asthma, eye inflammation, lung inflammation, pulmonary fibrosis, seborrheic dermatitis, pustular skin diseases, AIDS, allergic reactions, Alzheimer's disease, inflammatory diseases, atherosclerosis, bone diseases, breast cancer, cancer, cardiovascular disease, colon cancer, congenital defects, degenerative neurological disorders, dementia , dermatological disorders, diabetes, endocrine disorders, eye disorders, gastrointestinal disorders, genitourinary disorders, hearing disorders, blood disorders, hepatobiliary disorders, hypertension, immune disorders, infectious diseases, leukemia / lymphoma, lung cancer, metabolic disorders, neonatology, neurological disorders, neuromuscular disorders, obesity / eating disorders, plastic surgery disorders, parasitic diseases, perinatal disorders, prostate cancer, psychiatric disorders, pulmonary disorders, renal disorders, reproductive disorders, rheumatic diseases, stroke, surgical transplant disorders, vascular disorders, oral infections, periodontal disease, brain injury, trauma, and neuronal inflammation.

[0154] US20200010398 and US20200048177 describe novel monohydroxy, dihydroxy and trihydroxy derivatives derived from docosapentaenoic acid, and methods of treating or preventing inflammation associated with: arterial inflammation, arthritis, psoriasis, urticaria, vasculitis, asthma, ocular inflammation, lung inflammation, pulmonary fibrosis, seborrheic dermatitis, pustular dermatosis, cardiovascular disease, recruitment of neutrophils, leukocytes and / or cytokines, addiction, AIDS, alcohol-related disorders, allergies, Alzheimer's disease, arthritis, asthma, atherosclerosis, bone disease, breast cancer, cancer, cardiovascular disease, colon cancer, congenital defects, decision analysis, retirement Neurological disorders, Dementia, Dermatological disorders, Diabetes, Endocrine disorders, Eye disorders, Fetal and maternal medicine, Gastrointestinal disorders, Gene therapy, Genetic diagnostics, Genetics, Genitourinary disorders, Geriatrics, Growth and development, Hearing, Blood disorders, Hepatobiliary disorders, Hypertension, Infectious diseases, Leukemia, Lymphoma, Lung cancer, Metabolic disorders, Neonatology, Neurological disorders, Neuromuscular disorders, Obesity, Orthopedics, Parasitic diseases, Perinatal disorders, Pregnancy, Prostate cancer, Psychiatric disorders, Pulmonary disorders, Renal disorders, Reproduction, Rheumatic diseases, Stroke, Surgery, Transplantation, Vaccines, Vascular medicine, Trauma, Oral infections, Periodontal disease, Brain injury, Trauma and neuronal inflammation.

[0155] U.S. Patent Nos. 8,273,792, 9,364,454 and 9,782,379 describe protectins, including protectin D1 and protectin DX, and methods for treating or preventing airway inflammation and regulating blood glucose and inhibiting lipid-induced inflammation and other related inflammatory conditions such as insulin resistance, metabolic syndrome, type 2 diabetes, hypertension and cardiovascular disease.

[0156] In embodiments, the invention provides methods of treating diseases or conditions suitable for treatment with SPMs, including cancer, gastrointestinal diseases and conditions, inflammatory conditions, infectious diseases and conditions, metabolic diseases and conditions, neurological conditions, and pulmonary diseases and conditions. In embodiments, the method comprises administering to a subject in need of such therapy a pharmaceutical composition comprising a complex of SPMs and cyclodextrins as described herein, wherein the complex comprises an amount of SPMs effective to treat the disease or condition. In the context of the methods described herein, the term "treating" refers to regression, stabilization, or slowing of progression of the disease or condition being treated, or improvement or stabilization of one or more clinical symptoms associated with the disease or condition being treated.

[0157] cancer

[0158] The present disclosure provides methods for treating cancer in a subject in need of such treatment, the methods comprising administering to the subject a pharmaceutical composition comprising a complex described herein, wherein the complex comprises an amount of SPM effective to treat the cancer.

[0159] According to the methods described herein, administration can be used alone as a monotherapy, or in combination with one or more additional therapeutic agents. In embodiments, the combination therapy is adjuvant therapy, for example, as an adjuvant for chemotherapy, targeted therapy, immunotherapy, hormone therapy, gene therapy or microbial therapy. In embodiments, the combination therapy is an immune checkpoint inhibitor therapy. In embodiments, the combination therapy is an adjuvant or neoadjuvant therapy for surgery or radiotherapy. In embodiments, the composition can be used as a first-line, second-line, third-line or palliative therapy, alone or in combination with other therapies such as surgery, radiotherapy, chemotherapy, targeted therapy, immunotherapy, hormone therapy, gene therapy or microbial therapy. The term "therapy" refers to any method, regimen and / or agent that can be used to treat or control cancer.

[0160] In the context of the methods described herein, the term "treatment" may refer to the improvement or stabilization of one or more clinical symptoms associated with the cancer being treated. In embodiments, treatment results in the elimination of clinical symptoms of the cancer being treated, however, elimination is not required. In embodiments, the severity of the symptom is reduced. In the case of cancer, these symptoms may include clinical markers of severity or progression, including the size of solid tumors, the amount of growth factors or other biomarkers secreted by solid tumors, tumor angiogenesis, tumor metastasis or the number of metastases. Therefore, treating cancer according to the methods described herein may result in a decrease in tumor size or volume, also referred to as tumor regression; a decrease in the number of tumors; or a decrease in the number of metastatic lesions in other tissues or organs away from the primary tumor site. These symptoms may also include cancer cachexia or pain. Therefore, treating cancer according to the methods described herein may result in a reduction in cancer cachexia or pain, wherein the reduction in cachexia or pain is due to the administration of the complex described herein. In embodiments, "treating" cancer according to the methods described herein results in an increase in overall survival and / or an increase in progression-free survival in the subject being treated. In the context of the present disclosure, unless explicitly stated, the term "treatment" is not meant to include improvement or stabilization of clinical symptoms unrelated to the cancer being treated, but rather, for example, clinical symptoms related to the treatment modality, such as side effects or adverse events associated with another therapy such as radiation or chemotherapy. In addition, the term "treatment" does not include prevention, or administration of the SPM complex prior to the diagnosis of cancer in the subject being treated.

[0161] In embodiments, the invention provides methods for ameliorating adverse events or side effects of chemotherapy, targeted therapy, immunotherapy or radiotherapy, for example, gastrointestinal toxicity such as oral or gastrointestinal mucositis, colitis, enteritis, gastritis, nausea, vomiting and diarrhea, cardiovascular toxicity such as cardiomyopathy, myocardial ischemia, pericarditis, myocarditis, valvular heart disease, arrhythmia, congestive heart failure, hypertension, coronary, cerebrovascular and peripheral vascular events or congestive heart failure, skin toxicity such as pruritus or severe rash, hepatotoxicity, pulmonary toxicity such as pulmonary hypertension and pneumonia, endocrinopathy such as hypothyroidism, hyperthyroidism and hypophysitis, neurotoxicity such as Gram-negative Guillain-Barré syndrome, myasthenia gravis, posterior reversible encephalopathy syndrome, aseptic meningitis, enteric neuropathy, transverse myelitis, panencephalitis, autoimmune encephalitis and cranial nerve and peripheral neuropathy, or other side effects such as pancreatitis, acute kidney injury and cytokine release syndrome, the method comprising administering a complex described herein to a subject who is undergoing or will undergo chemotherapy, targeted therapy, immunotherapy or radiotherapy, wherein the resolvin is effective in reducing or preventing adverse events or side effects of chemotherapy, targeted therapy, immunotherapy or radiotherapy.

[0162] According to the methods described herein, a therapeutically effective amount of the SPM complex is administered to a subject in need of treatment. A therapeutically effective amount is an amount or dosage of the SPM active portion sufficient to treat cancer or to achieve a desired therapeutic outcome (e.g., improvement or stabilization of one or more biomarkers of disease progression or one or more clinical symptoms).

[0163] Gastrointestinal diseases and disorders

[0164] In an embodiment, the present disclosure provides a method of treating a gastrointestinal disease or disorder in a subject in need thereof by administering to the subject a pharmaceutical composition containing a complex described herein, wherein the complex comprises an amount of SPM effective to treat the gastrointestinal disease or disorder.

[0165] In embodiments, the gastrointestinal disease or disorder is selected from ulcerative colitis, Crohn's disease, proctitis, pouchitis, eosinophilic colitis, lymphocytic colitis, collagenous colitis, diversion colitis, chemical colitis, ischemic colitis, infectious colitis, pseudomembranous colitis, and indeterminate colitis. In embodiments, the gastrointestinal disease or disorder is selected from ulcerative colitis and Crohn's disease.

[0166] In embodiments, the gastrointestinal disease or disorder is selected from intestinal obstruction, chronic pancreatitis, colitis, colon cancer, congenital gastrointestinal anomalies, gastroschisis, high-output fistula, parenteral nutrition-related liver disease, postoperative ileus, postoperative enteritis, short bowel syndrome, and sporadic polyposis. In embodiments, the gastrointestinal disease or disorder is selected from eosinophilic esophagitis, Behcet's disease, irritable bowel syndrome, celiac disease, intestinal mucositis, NSAID enteropathy, intestinal infection, diverticular disease, diverticulitis, gastritis, pancreatitis, viral gastroenteritis, and Whipple's disease.

[0167] In embodiments, the gastrointestinal disease or disorder is postoperative intestinal inflammation, postoperative ileus, or a combination thereof. In embodiments, the gastrointestinal inflammatory disease or disorder is postoperative ileus (POI).

[0168] In preferred embodiments, the gastrointestinal disease or disorder is inflammatory bowel disease (referred to herein as "IBD"), a term given to two conditions, Crohn's disease and ulcerative colitis, both of which are characterized by chronic inflammation of the gastrointestinal tract. Crohn's disease can affect any part of the gastrointestinal tract, including the mouth and anus, but most commonly affects the lower gastrointestinal tract, especially the ileum and / or colon. Ulcerative colitis affects the colon and rectum. The terms "large intestine" and "colon" are used interchangeably in this disclosure.

[0169] Standard of care treatment for mild to moderate IBD aims to achieve and maintain remission by controlling inflammation in target tissues of the gastrointestinal tract. In adults, 5-aminosalicylate (5-ASA) can be used alone or in combination with corticosteroids to induce remission. Typically, corticosteroid therapy is administered for short-term symptom control, e.g., to treat acute flares and induce remission of disease symptoms. Long-term or maintenance therapy often includes the use of nonsteroidal medications, including 5-ASA and immunomodulators, such as azathioprine.

[0170] Standard of care treatment for moderate to severe IBD also aims to achieve and maintain remission by controlling inflammation in target tissues of the gastrointestinal tract. In adults, anti-TNF therapy with adalimumab, golimumab, or infliximab alone or in combination with corticosteroids, thiopurines, or 5-ASAs can be used to induce remission. For patients with moderate to severe IBD who have failed anti-TNF therapy, integrin receptor antagonists such as vedolizumab, sphingosine-1-phosphate receptor modulators such as ozanimod, or Janus kinase inhibitors such as tofacitinib can be used. Long-term or maintenance therapy generally consists of continuation of anti-TNF therapy, with or without thiopurines, or continuation of other therapies listed above.

[0171] In an embodiment, the present disclosure provides for treating IBD by administering to a subject a pharmaceutical composition comprising a complex as described herein, the pharmaceutical composition as a monotherapy or a combination therapy with a second IBD therapeutic agent. In an embodiment, the second IBD therapeutic agent is selected from corticosteroids, 5-ASA, azathioprine, anti-TNF therapy, integrin receptor antagonists, sphingosine-1-phosphate receptor modulators or Janus kinase inhibitors. In an embodiment, the present disclosure provides for treating IBD in a method for treating IBD with a combination of IBD maintenance therapy, optional 5-ASA, azathioprine, anti-TNF therapy, integrin receptor antagonists, sphingosine-1-phosphate receptor modulators or Janus kinase inhibitors. In an embodiment, the present disclosure provides for treating IBD in a method for treating IBD with a combination of any one or more of 5-ASA, azathioprine, corticosteroids, anti-TNF therapy, integrin receptor antagonists, sphingosine-1-phosphate receptor modulators or Janus kinase inhibitors. In an embodiment, the present disclosure provides a dual RvE1 magnesium dilysine for use in a method for treating IBD in combination with a corticosteroid. According to the aforementioned embodiment of the combined therapy of dual RvE1 magnesium dilysine and corticosteroids, corticosteroids can be selected from the group consisting of prednisone, prednisolone, budesonide, hydrocortisone and beclomethasone dipropionate, wherein the dosage form is an oral dosage form, or selected from hydrocortisone, prednisolone and budesonide, wherein the dosage form is suitable for rectal delivery, for example, in the form of an enema or suppository. According to the aforementioned embodiment of the combined therapy of dual RvE1 magnesium dilysine and anti-TNF therapy, integrin receptor antagonists, sphingosine-1-phosphate receptor modulators or Janus kinase inhibitors, the second IBD therapeutic agent can be selected from the group consisting of adalimumab, golimumab, infliximab, vedolizumab, ozanimod, etrasimod, tofacitinib or filgotinib.

[0172] Inflammatory diseases

[0173] Due to the ability of SPMs to mediate resolution of inflammation, and the ability of the compounds described herein to deliver therapeutically effective amounts of SPMs to tissues of subjects in need of treatment for inflammation, the compounds described herein are particularly useful for treating diseases and disorders with a significant inflammatory component.

[0174] In an embodiment, the disclosure provides a method of treating an inflammatory disease or condition in a subject, a pharmaceutical composition comprising a complex as described herein, wherein the complex comprises an amount of SPM effective to treat the inflammatory disease or condition.

[0175] In an embodiment, the inflammatory disease or condition is selected from the group consisting of acne, adipose tissue inflammation, allograft rejection, arthritis, bacterial infection, burn wounds, cheilitis, chronic pancreatitis, corneal wounds, dermatitis, diabetic wounds, dry eye syndrome, eczema, endometriosis, endotoxin shock, glossitis, cardiac ischemia, HSV-keratitis, ischemia-reperfusion injury, local aggressive periodontitis, Lyme arthritis, macular edema, oral mucositis, osteoarthritis, periodontitis, peritonitis, postoperative pain, postoperative cognitive decline, pruritus, psoriasis, pyoderma gangrenosum or hidradenitis suppurativa, retinopathy, rheumatoid arthritis, scleroderma, Sjögren's syndrome, steroid-induced lupus erythematosus, stomatitis, systemic inflammatory response syndrome, temporomandibular joint inflammation and vascular inflammation.

[0176] Infectious diseases and conditions caused by infectious agents

[0177] In an embodiment, the present disclosure provides a method of treating a disease or condition caused by an infectious agent (such as bacteria, fungi or viruses) in a subject in need thereof by administering to the subject a pharmaceutical composition containing a complex described herein, wherein the complex comprises an amount of SPM effective to treat the infectious disease or condition.

[0178] In embodiments, the disease or condition is a bacterial infection. In embodiments, the bacterial infection is bacterial pneumonia. In embodiments, the bacterial infection is an Escherichia coli infection. In embodiments, the bacterial infection is a Mycobacterium tuberculosis infection.

[0179] In embodiments, the disease or disorder is a yeast infection. In embodiments, the yeast infection is a Candida infection.

[0180] In embodiments, the disease or condition is sepsis. In embodiments, the sepsis is burn sepsis.

[0181] Metabolic diseases and conditions

[0182] In an embodiment, the disclosure provides a method of treating a metabolic disease or disorder in a human subject in need thereof by administering to the subject a pharmaceutical composition containing a complex described herein, wherein the complex comprises an amount of SPM effective to treat the metabolic disease or disorder.

[0183] In embodiments, the metabolic disease or condition is abnormal glucose metabolism manifested as diabetes, including type 2 diabetes or pre-diabetes, insulin resistance, abnormal lipid metabolism manifested as hypertriglyceridemia, i.e., elevated triglycerides, mixed dyslipidemia, hypercholesterolemia, fatty liver, and combined abnormalities of glucose and lipid metabolism manifested as obesity; or a dyslipidemia condition selected from hypertriglyceridemia, hypercholesterolemia, and mixed dyslipidemia.

[0184] In an embodiment, the metabolic disease or disorder is insulin resistance, mixed dyslipidemia, nonalcoholic steatohepatitis (NASH), type 2 diabetes, primary biliary syndrome, and primary sclerosing cholangitis.

[0185] According to the methods described herein, administration can be used alone as a monotherapy, or in combination with one or more additional therapeutic agents, including but not limited to antihyperlipidemic agents or antidiabetic agents. Antihyperlipidemic agents that can be used include HMG CoA enzyme inhibitors (e.g., statins), cholesterol absorption inhibitors, and cholesterol esterase transfer protein (CETP) inhibitors. In an embodiment, the antihyperlipidemic agent is selected from statins, cholesterol absorption inhibitors, CETP inhibitors, and pharmaceutically acceptable salts and prodrugs of any of the foregoing drugs.

[0186] Neurological disorders

[0187] In an embodiment, the present disclosure provides a method of treating a neurological disorder in a subject in need thereof by administering to the subject a pharmaceutical composition containing a complex described herein, wherein the complex comprises an amount of SPM effective to treat the neurological disorder.

[0188] In embodiments, neurological diseases and conditions that may be treated include, but are not limited to, Alzheimer's disease, peripheral nerve injury, amyotrophic lateral sclerosis, pain, and fibromyalgia. In embodiments, the neurological disease or condition is selected from the group consisting of postoperative delirium, acute postoperative pain, fibromyalgia, endometriosis, lower genital tract pain, vulvodynia, chronic low back pain, treatment or management of pain associated with osteoarthritis, diabetic peripheral neuropathy, and musculoskeletal injury or trauma. In the context of the methods described herein, the term "treating" may refer to the improvement or stabilization of one or more clinical symptoms associated with the disease or condition being treated.

[0189] In embodiments, the amount is effective to treat one or more symptoms of the neurological disorder.

[0190] In embodiments, the neurological disorder is a psychiatric disorder. In embodiments, the psychiatric disorder is selected from attention deficit hyperactivity disorder (ADHD) and depression. In embodiments, the neurological disease or disorder is postoperative cognitive dysfunction (POCD) or postoperative delirium.

[0191] The present disclosure also provides methods for treating or controlling pain. In an embodiment, the pain is nociceptive pain, and the method comprises administering to a subject in need of treatment for nociceptive pain a pharmaceutical composition comprising an effective amount of a compound described herein or a mixture thereof.

[0192] In embodiments, the present disclosure provides methods for treating or managing pain associated with inflammation, fibromyalgia, endometriosis, vulvodynia, osteoarthritis, diabetic peripheral neuropathy, and musculoskeletal injury or trauma.

[0193] In embodiments, the present disclosure also provides methods for treating or managing acute postoperative pain and chronic low back pain.

[0194] Lung diseases and conditions

[0195] In an embodiment, the present disclosure provides a method of treating a pulmonary disorder in a human subject in need thereof by administering to the subject a pharmaceutical composition containing a complex described herein, wherein the complex comprises an amount of SPM effective to treat the pulmonary disease or disorder.

[0196] In embodiments, pulmonary and vascular diseases and conditions that may be treated include, but are not limited to, asthma, lung inflammation, bronchopulmonary dysplasia (also known as infantile chronic lung disease), cystic fibrosis, allergic airway reactions, acute lung injury, acute respiratory distress syndrome, lung injury, idiopathic pulmonary fibrosis, bacterial pneumonia, cigarette smoke-induced lung inflammation, and vascular inflammation.

[0197] Example

[0198] Although SPMs have promising pharmacology for the treatment of inflammatory diseases and cancer, they are chemically unstable, in part because their cis double bonds tend to isomerize to a thermodynamically more favorable trans configuration, which has been found to be less potent than native SPM. It is therefore an object of the present invention to provide SPM complexes in which the stability of the SPM active moiety is significantly improved to enable their use in pharmaceutical compositions.

[0199] To improve the solid-state stability of resolvin E1 (RvE1), a representative resolvin of SPM with conjugated double bonds susceptible to isomerization, the present inventors evaluated various methods to protect RvE1 from exposure to oxygen, light and moisture without using analog modifications to the chemical structure of the SPM molecule. These methods include: (i) suspension of RVE1-MgLys in various lipid excipients, including peceol, maisine, labrasol ALF, Span 80, poloxamer Kollisov P124 and Tween 80; (ii) spray coating of RvE1-MgLys with hydroxypropyl cellulose in isopropanol at a ratio of 1:4 (w / w) onto microcrystalline cellulose beads with a hydroxypropyl cellulose seal coat using a Wurster fluidized bed process; (iii) adding antioxidants (0.5% w / w) including α-tocopherol, ascorbyl palmitate, propyl gallate, BHT and BHA to a methanol solution of RvE1-MgLys and freeze-drying to form a dry powder; and (iv) using cyclodextrins, including α-CD, β-CD, γ-CD, HP-β-CD and SBE-β-CD, to complex with RvE1-MgLys. Importantly, only the complex of RvE1-MgLys with cyclodextrin provided adequate stabilization of the active portion of RvE1. This finding is surprising, as it has been shown that the stabilizing effect of cyclodextrins can be positive or negative, depending on the specific interactions of the reactive portion of the guest molecule within the cyclodextrin cavity, and whether the complex is in the solid state or in solution. Given the unique conjugated double bond system of SPMs and their known vulnerability to isomerization, one skilled in the art would not a priori predict that they would be stabilized by one or more cyclodextrins under the specific test conditions described below.

[0200] In the experiments, RvE1-MgLys was complexed with various cyclodextrins in solution and then freeze-dried to form powders. The resulting compounds were characterized by NMR spectroscopy studies to confirm their novelty through complex formation, the strength of intramolecular interactions and the unique orientation of RvE1 within the CD cavity. The potency of these new compounds was determined by their improved chemical stability over RvE1-MgLys alone under accelerated conditions over a period of 16 weeks.

[0201] Preparation of complexes

[0202] Example 1: General method for preparing composites

[0203] The main techniques for complexing cyclodextrins with water-soluble drugs involve dissolving the guest and cyclodextrin in water or an organic solvent / water mixture and then isolating the complex by removing the solvent by crystallization, reduced pressure coevaporation, spray drying, or freeze drying. Based on the high solubility of RvE1-MgLys and the small amount prepared in this experiment, the freeze drying method was selected for the initial preparation of the complex. However, the other separation techniques mentioned above may be suitable for producing SPM complexes.

[0204] The following steps were used to prepare SPM complexes for NMR and stability studies. First, the calculated amount of RvE1-MgLys was mixed with the calculated amount of various cyclodextrins (see Table 2) in a 1:2 molar ratio and dissolved in 1400uL of D2O, stirred for 1 hour and filtered (0.22μm, MF-Millipore). For NMR studies, the filtrate was transferred to a standard 5mm glass NMR tube for testing. For stability studies, the filtrate was freeze-dried (VirTis BenchTop Pro freeze dryer) and transferred to an open test tube.

[0205] Table 2: Sample preparation of RvE1-MgLys cyclodextrin complex

[0206]

[0207] NMR interaction studies confirmed the complexation with cyclodextrin

[0208] 1H NMR and 2D ROESY NMR experiments were performed on a 600 MHz VarianDDR NMR spectrometer equipped with a 5 mm inverse detection gradient (IDPFG) probe at 298 K. Standard pulse sequences and processing procedures provided in VNMRJ 4.1 were used. 1H chemical shifts (δ) were referenced to the residual HOD peak present in D2O (δ = 4.7900 ppm).

[0209] 1H spectra were recorded from 16 scans with a 2 s relaxation delay applied. 2D Rotating Frame Overhauser Enhancement Spectroscopy (ROESY) was recorded from 8 scans / increment using a 2 s relaxation delay, 200 increments for the native cyclodextrin complex and 512 increments for the randomly substituted cyclodextrin complexes, the spin lock time was set to 300 ms in each case. Additional analysis was performed using heteronuclear single quantum coherence (HSQC) NMR to determine proton-carbon single bond correlations. The atomic numbering scheme for RvE1-MgLys and α-CD, β-CD, γ-CD, HP-β-CD and SBE-β-CD cyclodextrins used for NMR spectroscopy studies is as follows Figure 1 -3.

[0210] Example 2: NMR characterization and intramolecular interactions of RvE1-MgLys alone

[0211] Figure 4 The 1H-NMR spectrum of RvE1-MgLys with full signal assignment is shown. Signals of RvE1 are indicated by Roman numerals, signals of magnesium lysine are indicated by Greek letters, and possible impurities related to RvE1-MgLys are indicated by asterisks.

[0212] from Figure 4 In the 1H-NMR spectrum shown, three clear regions of RvE1 were identified: (1) an unsaturated region between δ1H=6.5 and 5 ppm; (2) a chiral center with secondary hydroxyl groups between δ1H=4.7 and 4 ppm; and (3) an aliphatic region between δ1H=2.5 and 0.7 ppm. The assignment of RvE1-MgLys was accomplished using further 2D ROESY and 13C NMR experiments.

[0213] To map the intramolecular interactions and any intermolecular interactions between the RvE1-MgLys components, 2DROESY analysis was performed ( Figure 5 ).

[0214] Example 3: NMR characterization and intermolecular interaction study of RvE1-MgLys complexed with α-cyclodextrin

[0215] like Figure 6 As shown, the NMR signals of the α-CD complex appear in a well-defined region of the spectrum. The anomeric proton appears between δ1H=5.0-5.2 ppm, while the other protons of the pyranose unit resonate between δ1H=3.5-4.0 ppm (commonly referred to as the core region).

[0216] Typically, complex formation causes shifts in the 1H-NMR spectra of the guest and host molecules, so that previously non-overlapping signals between guest and host may become overlapping (called multiplets), and overlapping signals may be partially or completely separated.

[0217] By comparing the 1H-NMR spectrum of the unsaturated region of RvE1-MgLys alone ( FIG. 7 a ) and the 1H-NMR spectrum of the unsaturated region of RvE1-MgLys in the α-CD complex ( FIG. 7 b ), the signal shifts associated with the complex formation of RvE1-MgLys and α-CD were shown. It was observed that multiple previously separated signals in the unsaturated region of RvE1-MgLys became overlapping multiplets (e.g., 8 / 16 and 11 / 17), while some initially overlapping signals shifted apart (7 / 15). Importantly, the shifts observed in the saturated region of RvE1-MgLys alone were minimal ( FIG. 8 ), compared to RvE1-MgLys in the α-CD complex, indicating that the interaction in this region of the RvE1 molecule is minimal.

[0218] 2D ROESY spectroscopy analysis was performed to provide detailed information on the possible host-guest interactions between RvE1-MgLys and α-CD ( Fig. 9 ). When the signal of α-CD appears in a well-defined spectral range, the possible area of ​​intermolecular interaction can be determined, as shown by the dashed rectangle, while the specific interaction between the guest and the host is emphasized by the solid rectangle. Using 2DROESY spectral analysis, intermolecular correlations between α-CD protons within the α-CD cavity (3' and 5') and at the main edge of the cavity (6') and RvE1-MgLys protons attached to unsaturated carbons (6-12) and one unsaturated carbon adjacent to it (5) were demonstrated. In addition, no cross peaks were detected between the amino acid chelate (MgLys) component of RvE1-MgLys and α-CD.

[0219] Fig.10 A zoomed-in portion of the intermolecular interactions is shown, which enables the exact mode of interaction to be determined. These spectra indicate that the protons of RvE1-MgLys that interact with α-CD are 5, 7, 8, 9, 10 and 11. Based on these data, Figure 23a provides a possible orientation of RvE1 within the α-CD cavity.

[0220] Example 4: NMR Characterization and Intermolecular Interaction Study of RvE1-MgLys Complexed with β-cyclodextrin

[0221] Fig.11 It was shown that the NMR signals of the β-CD complex appeared in a clear spectral region similar to that of the α-CD complex.

[0222] As shown in Figure 7, the signals of multiplexes 8 / 16 and 6 / 14 were completely and partially separated, respectively, while the separation of 7 / 15 was reduced compared to RvE1-MgLys alone, indicating the formation of a complex. Minimal signal shift was observed for the saturated portion of RvE1-Mg-Lys in the β-CD complex (Figure 8).

[0223] 2D ROESY spectroscopy analysis was also performed to provide detailed information on the possible host-guest interactions between RvE1-MgLys and β-CD ( Fig.12 ), the possible areas of intermolecular interaction are shown in the dashed rectangles, and the specific interactions are highlighted with solid rectangles. 2D ROESY analysis showed that some signals of RvE1-MgLys in the core region were spatially close to the internal protons (3' and 5') of β-CD, indicating the formation of a host-guest complex. The 2D ROESY spectrum ( Fig.13 ) to determine the exact site and pattern of inclusion, which showed a clear spatial proximity between the middle part of RvE1 (protons 8-15) and the inner side of the β-CD cavity (protons 3' and 5'). Based on these data, Figure 23b provides a possible orientation of RvE1 within the β-CD cavity.

[0224] Example 5: NMR Characterization and Intermolecular Interaction Study of RvE1-MgLys Complexed with γ-cyclodextrin

[0225] Fig.14 It was shown that the NMR signals of the γ-CD complex appeared in a clear spectral region similar to those of the α-CD complex and the β-CD complex.

[0226] As shown in Figure 7, the original overlapping signals of 8 / 16 and 6 / 14 were partially and completely separated, respectively, indicating the formation of a complex between RvE1-MgLys and the unsaturated region of γ-CD. As with the α-CD and β-CD complexes, the smallest signal shift was observed in the saturated part of RvE1-Mg-Lys in the γ-CD complex (Figure 8).

[0227] 2D ROESY spectroscopy of the γ-CD complex was performed to provide detailed information on possible host-guest interactions ( Fig.15 ). The analysis detected cross peaks between the RvE1-MgLys signal and the γ-CD core region in the designated region of intermolecular correlation shown by the dashed rectangle. The strongest cross peaks indicating correlation between the internal protons of γ-CD (3' and 5') and the unsaturated region signal of RvE1-MgLys are highlighted with solid rectangles. The 2D ROESY spectrum ( Fig.16) to determine the exact site and pattern of inclusion. The strongest cross peaks in γ-CD can be observed between the 9, 10 and 15 protons of RvE1-MgLys (located in the center of the molecule) and the 3', 5' and 6' protons of γ-CD. Additional signals are between the same protons of γ-CD and the 12 and 20 protons of RvE1-MgLys. Based on these data, Figure 23c provides a possible orientation of RvE1 within the γ-CD cavity.

[0228] Example 6: NMR Characterization of RvE1-MgLys Complexed with 2-Hydroxypropyl-β-cyclodextrin and Intermolecular Interactions effect Research

[0229] according to Fig.17 , the NMR signals of the HP-β-CD complex can be grouped into three independent regions. The anomeric protons appear between δ1H=5.0-5.2ppm. Because HP-β-CD is a randomly substituted derivative of natural CD, there are two types of anomeric signals in HP-β-CD: one represents the unsubstituted type, and the other (usually the one with a higher chemical shift) represents the substituted type. The HP-β-CD core was found to be between δ1H=3.5-4.2ppm, and the signals of the side chains (α' and β') were included. The methyl (γ') protons of the side chains established the third group of signals of HP-β-CD near δ1H=1.0-1.2ppm.

[0230] As shown in Figure 7, the slight signal change of RvE1-MgLys in the HP-β-CD complex compared with the spectrum of RvE1-MgLys alone indicates that the presence of HP-β-CD induces a chemical shift. The pattern is similar to that observed in the case of the β-CD system, with the largest difference being the partial separation of the 7 / 15 signal and the partial overlap of the 11 / 17 signal.

[0231] like Fig.18 As shown in the 2D ROESY spectrum analysis of the HP-β-CD complex in Figure 2, some cross peaks can be detected between the RvE1-MgLys signal and the HP-β-CD core region in the designated region of the intermolecular correlations indicated by the dashed rectangle. Further analysis of the zoomed-in portion of the 2D ROESY spectrum for intermolecular interactions shows that the strongest cross peaks are those of protons 9, 10, 12, and 18, with additional interactions detected for protons 6, 11, and 13 ( Fig.19 ). Based on these data, it is clear that the observed intermolecular interactions are localized at the 3', 5', and 6' protons of HP-β-CD in the interior, indicating the formation of a complex within the cavity, with a possible orientation shown in Figure 23d.

[0232] Example 7: NMR Characterization of RvE1-MgLys Complexed with Sulfobutyl Ether β-Cyclodextrin and Intermolecular Interactions Function research Research

[0233] according to Fig. 20 , the 1H NMR signals of the SBE-β-CD complex can also be grouped into three separate regions. The anomeric proton appears between δ1H=5.0-5.2ppm. Like HP-β-CD, SBE-β-CD has two types of anomeric signals: one represents the unsubstituted type, and the other (usually the one with a higher chemical shift) represents the substituted type. The SBE-β-CD core was found to be between δ1H=3.5-4.1ppm. The α' protons of the side chains overlap with the core region, and the remaining signals of the side chains include two separate regions, one between δ1H=2.9-3.0ppm, representing the δ' protons, and the other between δ1H=1.7-1.9ppm, including the intermediate protons β' and γ' of the side chains.

[0234] like Fig.21 As shown by the dashed and solid rectangles in the figure, the molecular interactions are shown between the RvE1-MgLys signal and the SBE-β-CD core region. Fig. 22 ) showed that there were cross peaks in the 3′, 5′, 6′, and α′ proton regions within the SBE-β-CD cavity, and their possible orientations are shown in Figure 23e.

[0235] Stability, pharmacokinetics and formulation studies of the complex

[0236] Example 8: Stabilization of RvE1-MgLys by the Cyclodextrins Investigated

[0237] The stability of RvE1-MgLys alone ("RVE1 control") and RvE1-MgLys complexed with cyclodextrin ("RVE1 CD complex") was evaluated. Samples of the test article were placed in open test tubes and maintained at 40°C and 75% relative humidity, exposed to air for 16 weeks. Quantitative demonstration of stability was determined using an HPLC analytical method. Briefly, HPLC analysis was performed on a HaloGuard column (Halo C18 4.6x150 mm, 2.7μm), which was mounted on an Agilent HPLC system equipped with a UV detector (272nm detection). The mobile phase consisted of a gradient between solution A (water) and solution B (methanol), both containing 100mM ammonium formate, pH 4.0. The gradient program for solution A was 95-0%, and the gradient program for solution B was 5-100%. The flow rate was 0.9mL / min over 50 minutes. Lack of stability was indicated by a decrease in the area percent purity (area %) of the single major peak of RvE1 (retention time 31.3 minutes) and the appearance of new HPLC peaks or an increase in the area % of pre-existing impurities. All HPLC measurements of RvE1 purity and degradant levels were calculated based on the area percent relative to the detectable components in the molecule, which included isomers and diastereomers of RvE1, impurities, and degradants induced by the test conditions.

[0238] At the initial time point (baseline), RvE1 eluted as a single major peak with an area % purity of 94% in the RvE1 control sample and 95.3% to 95.5% in the RvE1 CD complex sample (Table 3). After 16 weeks of exposure to the above test conditions, the RvE1 portion in the control group had been significantly degraded, as indicated by a decrease in the major peak to 80.7%. In contrast, under the same conditions, the RvE1 portion in the various RvE1 CD complexes was degraded to a lower extent.

[0239] As shown in Table 3, the stabilizing effect of complexing with cyclodextrin can be represented by the change in RvE1 purity from baseline to week 16, also known as the degradation rate. At week 16, the degradation rate of the RvE1 control was 14.1%, while the average degradation rate of the RvE1 CD complex was 3.9%. Therefore, for each cyclodextrin tested, the degradation rate of RvE1 in the RvE1CD complex was significantly reduced.

[0240] Table 3: RvE1 at Weeks 8 and 16 under accelerated conditions compared to RvE1 control RvE1 purity of CD complexes

[0241]

[0242]

[0243] Importantly, the superior stability of the RvE1 CD complex was also associated with the formation of much fewer degradation peaks greater than or equal to 0.2% or 1.0% at weeks 8 and 16 compared to the RvE1 control under the same conditions (Table 4). Notably, the RVE1 Mg-Lysγ-cyclodextrin complex showed the least number of degradation peaks greater than 0.2%. Overall, the enhanced stability of the RvE1 CD complex reduces the risk of potency loss and / or the appearance of unknown impurities with potential safety issues during the manufacturing, formulation, packaging, storage and distribution processes.

[0244] Table 4: Number of impurities ≥ 0.2% and 1.0% at Week 8 and Week 16 under accelerated conditions

[0245]

[0246] Example 9: Long-term stabilization of RvE1-MgLys and RvE1 sodium by cyclodextrin

[0247] The chemical stability of RvE1-Mg-Lys alone and RvE1 sodium alone ("Na-RvE1") and RvE1-Mg-Lys and RvE1 sodium complexed with cyclodextrins was evaluated over 24 weeks. Samples of the solid form of the test article were placed in open test tubes and maintained at 40°C and 75% relative humidity, exposed to air. Quantitative evidence of stability was determined using an HPLC analytical method as described above.

[0248] As shown in Table 5, at the initial time point (baseline), RvE1 eluted as a single major peak with an area % purity of 94.8% in the RvE1-MgLys sample alone ("RvE1 MgLys control"), 96.2% in the RvE1 sodium sample alone ("Na-RVE 1 control"), and 95.9% to 96.8% in the cyclodextrin complex samples formed by each RVE1 salt ("RVE1 CD complex"). After 24 weeks of exposure to the above test conditions, the RvE1 portion of the RvE1-MgLys control group had been significantly degraded, as indicated by a decrease to 79.1% of the major peak (consistent with the results shown in Table 3), and the RvE1 portion of the Na-RvE1 control group had been completely degraded to 0%. In contrast, under the same conditions, the RvE1 portion of the RvE1 CD complex was degraded to a lower extent. Importantly, the results shown in Table 5 demonstrate that the superior stability achieved by complexing with cyclodextrin can be achieved with different initial forms of the RvE1 moiety, in this example with RvE1 in the form of the sodium or magnesium lysine salt.

[0249] At week 24, the degradation rates of the RvE1-MgLys control and Na-RvE1 control were 16.6% and 100%, respectively, whereas the degradation rates of the RvE1-MgLysγ-cyclodextrin complex were 0%, the RvE1-MgLys HP-β-CD complex were 3.4%, and the Na-RVE1 HP-β-CD complex were 4.3%, which were consistent with the results shown in Table 3.

[0250] Table 5: -SPM at Week 24 under Accelerated Conditions RvE1 purity of CD complexes

[0251]

[0252] Example 10: Stabilization of RvE1 Salt in Solution by Cyclodextrin

[0253] The chemical stability of RvE1-MgLys or Na-RvE1 alone and in complex with cyclodextrin in solution was evaluated. Samples of the test article at a concentration of 2 mg / ml RVE1 dissolved in deionized water were placed in open test tubes and maintained at 40°C and 75% relative humidity, exposed to air for more than 16 weeks. Quantitative evidence of stability was determined using an HPLC analytical method as described above.

[0254] As shown in Table 6, at the initial time point (baseline), RvE1 eluted as a single major peak in all samples, with an area % purity of 94.8% to 96.2% in the RvE1-MgLys alone ("RvE1-MgLys control") and Na-RvE1 alone ("Na-RVE1 control") samples, and an area % purity of 95.9% to 96.8% in the RvE1 cyclodextrin complex ("RvE1CD complex") samples. After 16 weeks of exposure to the above test conditions, the RvE1 portion in both control groups had been significantly degraded, as indicated by a decrease in the major peak in the RvE1-MgLys control group to 51.5% and a decrease in the major peak in the Na-RvE1 control group to 18.6%. In contrast, under the same conditions, the RvE1 portion in the RvE1 CD complex was degraded to a lower extent.

[0255] Overall, the excellent stability of the RvE1 CD complex in solution is unexpected, as there are examples of cyclodextrin compounds that have excellent stability as a solid but poor stability in solution, or high stability as a solution but poor stability as a solid (Hamada Y et al., ChemPharm Bull (Tokyo). 1975; 23(6): 1205-1211; Uekama 2006; Popielec 2017). In addition, the results of RvE1 Mg-Lys and Na-RvE1 complexed with cyclodextrins indicate that enhanced stability can be achieved with different forms of the RvE1 moiety, and in this example, when the two RvE1 salts are not complexed with cyclodextrins, they have very different stability characteristics under accelerated conditions.

[0256] Table 6: RvE1 purity of SPM complexes in solution under accelerated conditions

[0257]

[0258]

[0259] Example 11: Solid-state physical stability of RvE1 MgLys γ-cyclodextrin complex

[0260] The solid state physical stability of RvE1 MgLys alone ("RVE1 control") and the RvE1-MgLys γ-cyclodextrin complex ("RVE1 complex") was evaluated. Fig.24A -D). Samples of the test article in solid form are placed in an open test tube for more than 168 hours under ambient conditions of a temperature of 18-22°C and a relative humidity between 52% and 72%. The solid state physical stability is assessed visually based on whether the substance retains its solid powder form or transforms into a liquid form.

[0261] At the initial time point (baseline) ( Fig.24A ), RvE1 control and RvE1 complex samples were in powder form. Fig. 24B ), while the RvE1 control sample began to aggregate and turn darker in color, the RvE1 control and RvE1 composite samples remained in powder form. Fig.24C -D), the RvE1 control sample has turned into a gel-like substance, while the sample of the RvE1 complex retains its form as a solid, off-white powder.

[0262] Example 12: Oral Pharmacokinetics of RvE1 Control and RvE1 CD Complex

[0263] To establish the comparative oral pharmacokinetics ("PK") of RvE1 MgLys alone ("RvE1 control") and RvE1 MgLys complexed with γ-cyclodextrin ("RVE1 complex"), a study was conducted to investigate the exposure of RVE1 in plasma after administration of a single dose of RVE1 control or RVE1 complex (4 mg / kg RVE1) by oral gavage at various time points for up to 8 hours. Quantitative measurement of RvE1 in plasma was determined by HPLC-MS / MS performed on a Halo Guard column (Halo C18 4.6×150 mm, 2.7 μm) mounted on an Agilent HPLC system equipped with a UV detector (272 nm detection).

[0264] Sixty-five (65) male C57Bl6 mice were randomized at 8 weeks of age into three experimental groups: baseline control ("Baseline", n = 5), RvE1 control (n = 30), and RvE1 complex (n = 30). Prior to administration of the test article, mice in the baseline group were sacrificed and blood was drawn to determine endogenous levels of RvE1 in plasma. The remaining 60 mice received a single dose of RvE1 control or RvE1 complex. Five (5) mice in each group were sacrificed at each of the following time points to determine the RvE1 concentration versus time curve: 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, and 8 hours after dosing. At the time of sacrifice, blood was drawn into EDTA-treated tubes and immediately frozen.

[0265] As shown in the PK profiles and PK indicators summarized in Table 7, the plasma PK of the RvE1 control group and the RvE1 complex group were comparable in terms of coverage area (AUC, ng / ml*hr), Cmax (ng / ml) and Tmax (min). In summary, these data indicate that oral administration of RvE1 complexes can deliver comparable levels of RvE1 to the systemic circulation compared to the RvE1 control, thus supporting the utility of cyclodextrins for RvE1 drug development.

[0266] Table 7: Oral pharmacokinetics of RvE1 control compared to RvE1 complex

[0267]

[0268] Example 13: RvE1MgLysγ-cyclodextrin complex in direct admixture with pharmaceutical excipients system Feasibility study

[0269] To determine the feasibility of using RvE1 salt complexed with cyclodextrin to prepare oral tablets, a study was conducted to test the flow and compression properties of a direct mixture ("direct mixture") consisting of RvE1 MgLysγ-cyclodextrin complex ("RvE1 complex") and standard pharmaceutical excipients shown in Table 8, without granulation to improve flow properties.

[0270] Table 8: Direct mixture of RvE1 complexes with standard pharmaceutical excipients

[0271]

[0272] In the flow portion of the study, a direct blend without granulation was first evaluated to determine the particle size distribution ("PSD") using screens ranging from #40 (coarse) to #325 (fine), corresponding to mesh diameters ranging from 400 to 44 microns. As shown in Table 9, approximately 75% of the direct blend particles were greater than or equal to 100 microns (coarser) and approximately 25% were less than 100 microns in diameter (finer). Once the particle size distribution of the direct blend was determined, flow experiments were conducted using an orifice with a diameter of 16 mm. In this portion of the study, the direct blend without additional granulation exhibited sufficient flow to pass through a 16 mm orifice, which was sufficient to achieve gravity feeding during compression on a rotary press or direct filling into capsules.

[0273] Table 9: Particle size distribution of direct blends

[0274]

[0275]

[0276] In the compression part of the study, the direct blend was compressed on a rotary press with oval tooling (0.5457 cm x 2852 cm) at a pressure of approximately 100 bar to produce tablets approximately 5.8 mm thick (see Fig.25A -B). Tablets with a target total weight of 500 mg per tablet were prepared into two groups: a low hardness group and a high hardness group (18 and 30 kgf, respectively). As shown in Table 10, the tablets in both groups had minimal weight variation and showed hardness, friability, and disintegration time properties consistent with the target specifications.

[0277] Table 10: Compression study of direct mixtures of GCD complexes

[0278] parameter Low (18kp) High (30kp) Individual weight (mg) 503-507 498-501 Thickness(mm) 7.3 6.3 Hardness(kp) 17-20 27-30 Friability (%) 0.1 0.0 Disintegration 1min 12sec 5min 23sec

[0279] Equivalent

[0280] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the appended claims.

[0281] All references cited herein are incorporated by reference in their entirety and for all purposes to the same extent as if each individual publication or patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety for each purpose.

[0282] The present invention is not limited by the scope of the specific embodiments described herein. In fact, various modifications of the present invention, in addition to those described herein, will become apparent to those skilled in the art based on the foregoing description and the accompanying drawings. These modifications are intended to fall within the scope of the appended claims.

Claims

1. A complex of a specialized resolution promoting mediator (SPM) or a salt, ester or amide thereof with a cyclodextrin (CD), wherein the SPM is a hydroxylated polyunsaturated fatty acid derived from arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid or docosapentaenoic acid, having a length of 20 or 22 carbon atoms and having 4, 5 or 6 conjugated double bonds placed in a diene, triene or tetraene system or a combination thereof.

2. The complex of claim 1, wherein the SPM is selected from the group consisting of RvE1, RvE2, RvE3, RvE4, AT-RvE1, AT-RvE2, AT-RvE3, RvD1, RvD2, RvD3, RvD4, RvD5, RvD6, AT-RvD1, AT-RvD2, AT-RvD3, AT-RvD4, AT-RvD5, AT-RvD6, RvT1, RvT2, RvT3, RvT4, LXA4, LXB4, AT-LXA4, AT-LXB4, PD1, PDX, AT-PD1, MaR2, PD1 n-3DPA or MaR1 n-3DPA or a group consisting of a salt, ester or amide thereof; optionally, wherein the SPM is RvE1, AT-RvE1, RvD1, AT-RvD1, RvD2, AT-RvD1, AT-RvD2, LxA4 or AT-LxA4 or a salt, ester or amide thereof.

3. The complex according to claim 1, wherein the cyclodextrin is selected from α-cyclodextrin (α-CD), β-cyclodextrin (β-CD), γ-cyclodextrin (γ-CD), 2-hydroxypropyl-β-cyclodextrin (HP-β-CD), sulfobutyl ether β-cyclodextrin (SBE-β-CD); 2-hydroxypropyl-α-cyclodextrin; randomly methylated β-cyclodextrin; 2-O-methyl-β-cyclodextrin; 2,6-di-O-methyl-β-cyclodextrin; heptathiol (2,3,6-tri-O-methyl)-β-cyclodextrin; carboxymethyl-β-cyclodextrin; carboxyethyl-β-cyclodextrin; hydroxyethyl-β-cyclodextrin; maltosyl-β-cyclodextrin -cyclodextrin; 3,6-(N,N,N-trimethylammonium)propyl-β-cyclodextrin; acetyl-β-cyclodextrin; 2,6-di-O-methyl-γ-cyclodextrin; 2-hydroxypropyl-γ-cyclodextrin; or sulfobutyl ether γ-cyclodextrin; optionally, wherein the cyclodextrin is α-cyclodextrin (α-CD), β-cyclodextrin (β-CD), γ-cyclodextrin (γ-CD), 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) or sulfobutyl ether β-cyclodextrin (SBE-β-CD); optionally, wherein the cyclodextrin is γ-cyclodextrin (γ-CD) or 2-hydroxypropyl-β-cyclodextrin (HP-β-CD).

4. The complex according to any one of claims 1 to 3, wherein the SPM is present in an amount of weight percentage relative to the total weight of the complex (including only the weight of the SPM and cyclodextrin molecules) ranging from 1.5% to 35%, 2% to 30%, 3% to 25% or 4% to 20%.

5. The complex according to any one of claims 1 to 3, wherein the SPM and cyclodextrin are present in a molar ratio ranging from 2:1 to 1:4, or optionally 1:1 to 1:3, or 1:1.5 to 1:2.5, or 1:

2.

6. The complex according to any one of claims 1 to 5, wherein the SPM is in the form of a salt.

7. The complex according to claim 6, wherein the SPM is in the form of a sodium salt, a potassium salt, a calcium salt, a zinc salt or a magnesium salt, optionally wherein the SPM is in the form of a magnesium salt of dilysine. The complex according to claim 7 , wherein the SPM is RvE1 sodium salt.

9. The complex according to claim 6, wherein the SPM is in the form of a salt described by formula I: in M is selected from magnesium (Mg 2+ ), calcium (Ca 2+ ) and zinc (Zn 2+ ) of divalent metals; A 1 and A 2 Each is the same SPM molecule; R 1 and R 2 Each independently is a C1-C 10 Alkyl; and X 1 and X 2 Each is independently H or CO-Z and Z is a peptide comprising 1 to 5 amino acids.

10. The complex according to claim 9, wherein M is selected from magnesium (Mg 2+ ) or calcium (Ca 2+ ).

11. The complex according to claim 9 or 10, wherein R 1 and R 2 Each independently is -(CH2)3–Y 1 and -(CH2)4–Y 2 , and Y 1 and Y 2 Each is selected from the group consisting of a positively charged primary amine, a positively charged secondary amine, a positively charged tertiary amine, and a positively charged guanidine.

12. The complex according to any one of claims 9 to 11, wherein X 1 and X 2 Each is H.

13. The complex according to any one of claims 9 to 12, wherein R 1 and R 2 Each is -(CH2)4–Y 2 And Y 2 Yes - NH3 + .

14. The complex according to claim 9, wherein M is magnesium (Mg 2+ ), R 1 and R 2 Each is -(CH2)4–Y 2 And Y 2 Yes - NH3 + , X 1 and X 2 Each is H, and A 1 and A 2 is RvE1, and the SPM of the complex is referred to as RvE1 magnesium dilysinate, or "RvE1MgLys".

15. The complex according to claim 9, wherein M is magnesium (Mg 2+ ), R 1 and R 2 Each is -(CH2)4–Y 2 And Y 2 Yes - NH3 + , X 1 and X 2 Each is H, and A 1 and A 2 is RvD1, and the SPM of the complex is referred to as RvD1 magnesium dilysinate, or "RvD1MgLys".

16. The complex according to claim 9, wherein M is magnesium (Mg 2+ ), R 1 and R 2 Each is -(CH2)4–Y 2 And Y 2 Yes - NH3 + , X 1 and X 2 Each is H, and A 1 and A 2 is RvD2, and the SPM of the complex is referred to as RvD2 magnesium dilysinate, or "RvD2MgLys".

17. The complex according to claim 9, wherein M is magnesium (Mg 2+ ), R 1 and R 2 Each is -(CH2)4–Y 2 And Y 2 Yes - NH3 + , X 1 and X 2 Each is H, and A 1 and A 2 is LxA4, and the SPM of the complex is referred to as LxA4 magnesium dilysinate, or "LxA4MgLys".

18. The complex according to any one of claims 9 to 17, wherein the cyclodextrin component of the complex is γ-cyclodextrin (γ-CD) or 2-hydroxypropyl-β-cyclodextrin (HP-β-CD).

19. The complex of claim 18, wherein the SPM component is a sodium salt, a potassium salt, a calcium salt, a zinc salt or a magnesium salt of RvE1.

20. A complex of RvE1 MgLys and γ-cyclodextrin (γ-CD).

21. A complex of RvE1 MgLys and 2-hydroxypropyl-β-cyclodextrin (HP-β-CD).

22. A complex of sodium salt of RvE1 and γ-cyclodextrin (γ-CD).

23. A complex of sodium salt of RvE1 and 2-hydroxypropyl-β-cyclodextrin (HP-β-CD).

24. A pharmaceutical composition comprising the complex according to any one of claims 18 to 23, and one or more pharmaceutically acceptable carriers and / or excipients.

25. The composition of claim 24, wherein the composition is formulated as an oral, pulmonary, rectal, parenteral, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, inhalation, buccal, sublingual, intrapleural, intrathecal, or intranasal dosage form; optionally, wherein the composition is formulated as an oral or parenteral dosage form.

26. A composition according to claim 24 or 25 for use in a method of treating a gastrointestinal inflammatory disease or condition selected from inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, proctitis, pouchitis, pouch Crohn's disease, eosinophilic colitis, lymphocytic colitis, collagenous colitis, diversion colitis, chemical colitis, ischemic colitis, eosinophilic esophagitis, Behcet's disease, irritable bowel syndrome, celiac disease, intestinal mucositis, diverticulitis and short bowel syndrome, optionally wherein the inflammatory disease or condition is ulcerative colitis, Crohn's disease or pouchitis.

27. A composition according to claim 24 or 25, for use in a method of treating cancer, optionally wherein the cancer is a solid tumor cancer, such as brain cancer, breast cancer, bladder cancer, colorectal cancer, esophageal cancer, gastric cancer, head and neck cancer, liver cancer, lung cancer, melanoma, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma or sarcoma, optionally wherein the cancer is colorectal cancer, gastric cancer, liver cancer, lung cancer, melanoma, ovarian cancer or pancreatic cancer.

28. The composition of claim 24 or 25, wherein the pharmaceutical composition is administered orally or parenterally, optionally wherein the parenteral administration is subcutaneous, intraperitoneal, intramuscular or intravenous; optionally wherein the pharmaceutical composition is administered sublingually or by inhalation.

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