CYCLODEXTRIN COMPLEXES OF SPECIALIZED PRORESOLVING MEDIATORS

Abstract

The present invention provides complexes of specific pro-degradation mediators (SPMs), or salts, esters, or amides thereof, and cyclodextrin (CD), where the SPMs are hydroxylated polyunsaturated fatty acids derived from arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, or docosapentaenoic acid, 20 or 22 carbon atoms in length and with 4, 5, or 6 conjugated double bonds arranged in a diene, triene, or tetraene system, or combinations thereof; and related compositions and methods.

Description

[Technical field]

[0001] This invention relates to inclusion complexes of cyclodextrins and special pro-resolution mediators (herein referred to as "SPMs"), including resolvins, protectins, lipoxins, and maresins, and their use in therapy. [Background technology]

[0002]

[0002] Human inflammatory responses have two phases: initiation and resolution. At the cellular level, initiation is a protective response characterized by the production and release of proinflammatory mediators that activate the innate immune response, resulting in the influx of polymorphonuclear cells (PMNs) from the cellular compartment that kills and clears foreign invaders and damaged cells. This initiation phase is an active process driven by metabolites of arachidonic acid, including prostaglandins, which are chemoattractants for eosinophils, neutrophils, and monocytes, and leukotrienes, which induce leukocyte adhesion, chemotaxis, and aggregation.

[0003]

[0003] During peak inflammation, as the inflammatory stimuli are eliminated, the resolution phase begins, characterized by a reduction in proinflammatory signals and a switch from the production and release of proinflammatory mediators to the production and release of special proresolution mediators (SPMs), including resolvins, protectins, lipoxins, and maresins. SPMs are hydroxylated polyunsaturated fatty acids derived from arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, or docosapentaenoic acid, 20 or 22 carbon atoms in length and with 4, 5, or 6 double bonds arranged in diene, triene, and / or tetraene systems. As endogenous pro-resolving counterparts to prostaglandins and leukotrienes, SPMs can halt PMN infiltration and stimulate the recruitment and activation of monocytes and pro-resolving macrophages to drive the degradative phase by clearing tissues of cellular debris in a non-inflammatory manner, restoring tissue homeostasis (Serhan CN et al. Cold Spring Harb Perspect Biol 2015;7:a016311).

[0004]

[0004] Unresolved inflammation is an important but underappreciated driver of many chronic diseases and disorders. Therefore, the ability to resolve excessive inflammation is of paramount importance 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]

[0005] In vitro studies of SPMs show potent effects on inflammation and other markers at nanomolar concentrations. In vivo studies of SPMs in a wide range of inflammatory disease and tumor models show effects on disease progression and pathology. These data provide a scientific rationale for the therapeutic administration of SPMs 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, Gilliganmm et al., J Exp Med. 2018;215(1):115-140; Gilliganmm, Gartung A, Sulciner ML, Proc Natl Acad Sci US A. 2019;116(13):6292-6297).

[0006]

[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, in therapeutically effective amounts to target tissues. The present invention addresses this need by providing SPMs stabilized in complexes with cyclodextrins. Summary of the Invention

[0007]

[0007] The present invention relates to complexes of SPM and cyclodextrin (herein referred to as "SPM complexes") for use in the treatment of inflammatory diseases and cancer. The SPM complexes and methods described herein are based on the unexpected finding of improved chemical and physical stability of the SPM moiety when complexed with cyclodextrin and exposed to air at elevated temperature and humidity, thus increasing the ability to manufacture, formulate, package, evaluate, distribute and utilize pharmaceutical compositions of SPM to prevent and treat inflammatory diseases and cancer. More specifically, the present invention relates to SPM complexes and their use in pharmaceutical compositions, where the stability of the SPM active moiety is significantly improved.

[0008]

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

[0009]

[0009] In an embodiment, the SPM includes 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, MaR 1 , and MaR 2 , PD1 n-3 DPA Or MaR 1 n-3 DPAor a salt, ester, or amide thereof; optionally, 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.

[0010] In an embodiment, the cyclodextrin is 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;Heptakis(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, the cyclodextrin is α-cyclodextrin (α-CD), β-cyclodextrin (β-CD), γ-cyclodextrin (γ-CD), 2-hydroxypropyl-β-cyclodextrin (HP-β-CD), or sulfobutylated β-cyclodextrin (SBE-β-CD); optionally, the cyclodextrin is γ-cyclodextrin (γ-CD) or 2-hydroxypropyl-β-cyclodextrin (HP-β-CD).

[0011]

[0011] In embodiments, the SPM is present in a weight percentage relative to the total weight of the complex (including only the weight of the SPM and cyclodextrin molecules) in the range of 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 from 1:1 to 1:3, or from 1:1.5 to 1:2.5, or 1:2.

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

[0014] In an embodiment, the SPM has formula I:

[0015] [ka]

[0016] In the form of a salt described below. [In the formula, M is magnesium (Mg 2+ ), Calcium (Ca 2+ ), and zinc (Zn 2+ ) is a divalent metal selected from A 1 and A 2 are each the same SPM molecule; R 1 and R 2 Each independently represents a C1 to C6 alkyl group having at least one basic functional group. 10 is alkyl; X 1 and X 2 are each independently H or CO-Z, and Z is a peptide containing 1 to 5 amino acids. In an embodiment of Formula I, M is magnesium (Mg 2+ ) or calcium (Ca 2+ ) is selected.

[0017] In an embodiment of formula I, R1 and R 2 are each independently -(CH2)3-Y 1 , and (CH2)4-Y 2 and Y 1 and Y 2 are each selected from a positively charged primary amine, a positively charged secondary amine, a positively charged tertiary amine, and a positively charged guanidine.

[0018] In the embodiment of formula I, X 1 and X 2 are each H. In an embodiment of formula I, R 1 and R 2 are -(CH2)4-Y 2 and Y 2 -NH3 + It is.

[0019] In an embodiment of Formula I, M is magnesium (Mg 2+ ) and R 1 and R 2 are -(CH2)4-Y 2 and Y 2 -NH3 + and X 1 and X 2 are H and A 1 and A 2 is RvE1, and the SPM of the complex is also called RvE1 magnesium dilysinate, or "RvE1 MgLys."

[0020] In an embodiment of Formula I, M is magnesium (Mg 2+ ) and R 1 and R 2 are -(CH2)4-Y 2 and Y 2 -NH3 + and X 1 and X 2 are H and A 1 and A 2 is RvD1, and the SPM of the complex is also called RvD1 magnesium dilysinate, or "RvD1 MgLys."

[0021] In an embodiment of Formula I, M is magnesium (Mg 2+ ) and R 1 and R 2 are -(CH2)4-Y 2 and Y 2 -NH3 + and X 1 and X 2 are H and A 1 and A 2 is RvD2, and the SPM of the complex is also called RvD2 magnesium dilysinate, or "RvD2 MgLys."

[0022] In an embodiment of Formula I, M is magnesium (Mg 2+ ) and R 1 and R 2 are -(CH2)4-Y 2 and Y 2 -NH3 + and X 1 and X 2 are H and A 1 and A 2 is LxA4, and the SPM of the complex is also called LxA4 magnesium dilysinate, or "LxA4 MgLys."

[0023] In an embodiment of a complex including an SPM of formula I, the cyclodextrin component of the complex is γ-cyclodextrin (γ-CD) or 2-hydroxypropyl-β-cyclodextrin (HP-β-CD).

[0024]

[0024] The invention also provides a method for preparing the complexes described herein. In an embodiment, the method includes combining an amount of SPM or a salt thereof with an amount of cyclodextrin to form a mixture; dissolving the mixture in water; and isolating the complex by a process including filtering to obtain a filtrate, followed by one or more of freeze-drying, crystallization, co-evaporation, or spray-drying the filtrate. In an embodiment, the dissolving step is performed under nitrogen, for example, by stirring under nitrogen for 1-3 hours. Also provided is a complex prepared by the method described above.

[0025]

[0025] In an embodiment, the SPM component of the complex described herein has a chemical purity of at least 88% as determined by high pressure liquid chromatography (HPLC) under conditions of 40°C and 75% relative humidity while exposed to air for at least 8 weeks, or 12 weeks or 16 weeks.

[0026] 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.

[0027] In embodiments, the number of degradation peaks of the SPM components of complexes present at 1.0% or more is less than 8, less than 6, or less than 4, or no more than 2, as determined by HPLC. In embodiments, the number of degradation peaks of the SPM components of complexes present at 0.2% or more is less than 16, optionally less than 14, or less than 12, or less than 10, or no more than 8, as determined by HPLC.

[0027]

[0028] The present invention also provides a complex prepared by the methods described herein.

[0029] The present invention also provides compositions comprising the conjugates described herein and one or more pharma- ceutically acceptable carriers and / or excipients.

[0028]

[0030] In aspects, the compositions are formulated as oral, pulmonary, rectal, parenteral, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, inhalation, buccal, sublingual, intrapleural, intrathecal, or intranasal dosage forms; optionally, the compositions are formulated as oral or parenteral dosage forms.

[0029]

[0031] In an aspect, the compositions described herein are for use in a method of treating an inflammatory disease or disorder of the gastrointestinal tract selected from inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, proctitis, pouchitis, fossa 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 disorder is ulcerative colitis, Crohn's disease, or pouchitis.

[0030]

[0032] In aspects, the compositions described herein are for use in a method of treating cancer, optionally wherein the cancer is a solid tumor cancer, e.g., 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.

[0031]

[0033] In aspects, the compositions are administered orally or parenterally, optionally, parenteral administration is subcutaneous, intraperitoneal, intramuscular, or intravenous; optionally, the pharmaceutical compositions are administered sublingually or by inhalation. [Brief description of the drawings]

[0032] [Figure 1]

[0034] Structure and atom number assignment of RvE1-MgLys. [Diagram 2]

[0035] Structures and atom number assignments for α-cyclodextrin (α-CD, A), β-cyclodextrin (β-CD, B), and γ-cyclodextrin (γ-CD, C). [Diagram 3]

[0036] Structures and atom number assignments of 2-hydroxypropyl-β-cyclodextrin (HP-β-CD, A) and sulfobutyl ether β-cyclodextrin (SBE-β-CD, B). [Figure 4]

[0037] 1H-NMR spectrum of RvE1-MgLys with complete assignments without complexation to cyclodextrin. [Diagram 5]

[0038] 2D ROESY spectrum of RvE1-MgLys without complexation to cyclodextrin. [Figure 6]

[0039] 1H-NMR spectrum of RvE1-MgLys complexed with α-CD with complete assignments. [Figure 7]

[0040] Stacked partial 1H-NMR spectra of the unsaturated region of RvE1-MgLys alone (A, first from the bottom) and the unsaturated region complexed with α-CD, β-CD, γ-CD, HP-β-CD, and SBE-β-CD (B–F, respectively). [Figure 8]

[0041] Stacked partial 1H-NMR spectra of the saturated region of RvE1-MgLys alone and the saturated region complexed with α-CD, β-CD, γ-CD, HP-β-CD, and SBE-β-CD, A to F, respectively. [Figure 9]

[0042] 2D ROESY spectrum of RvE1-MgLys complexed with α-CD with complete assignment. [Figure 10]

[0043] Partial 2D ROESY spectrum of RvE1-MgLys complexed with α-CD. [Figure 11]

[0044] 1H-NMR spectrum of RvE1-MgLys complexed with β-CD, including integrations and complete assignments. [Figure 12]

[0045] 2D ROESY spectrum of RvE1-MgLys complexed with β-CD with complete assignments. [Figure 13]

[0046] Zoomed-in partial 2D ROESY spectrum of RvE1-MgLys complexed with β-CD. [Figure 14]

[0047] 1H-NMR spectrum of RvE1-MgLys complexed with γ-CD, including integration and complete assignments. [Figure 15]

[0048] 2D ROESY spectrum of RvE1-MgLys complexed with γ-CD with complete assignments. [Figure 16]

[0049] Zoomed-in partial 2D ROESY spectrum of RvE1-MgLys complexed with γ-CD. [Figure 17]

[0050] 1H-NMR spectrum of RvE1-MgLys complexed with HP-β-CD, including integrations and complete assignments. [Figure 18]

[0051] 2D ROESY spectrum of RvE1-MgLys complexed with HP-β-CD with complete assignments. [Figure 19]

[0052] Zoomed-in partial 2D ROESY spectrum of RvE1-MgLys complexed with HP-β-CD. [Figure 20]

[0053] 1H-NMR spectrum of RvE1-MgLys complexed with SBE-β-CD, including integrations and complete assignments. [Figure 21]

[0054] 2D ROESY spectrum of RvE1-MgLys complexed with SBE-β-CD with complete assignments. [Figure 22]

[0055] Zoomed in partial 2D ROESY spectrum of RvE1-MgLys complexed with SBE-β-CD. [Figure 23]

[0056] Schematic diagrams showing the molecular orientation of RvE1 in the cavities of α-CD, β-CD, γ-CD, HP-β-CD, and SBE-β-CD, A–E, respectively. [Figure 24]

[0057] Photographs of samples of RvE1 MgLys alone and RvE1 MgLys complexed with γ-CD exposed to air under ambient conditions at baseline, 8 h, 24 h, and 168 h. [Diagram 25]

[0058] Photographs of direct blends of RvE1 MgLys complexed with γ-CD with standard pharmaceutical excipients without further granulation before tableting (A) and after compression into core tablets (B). [Figure 26]

[0059] Schematic diagram of the complex of RvE1 magnesium dilysinate (RvE1 MgLys) and γ-cyclodextrin (γ-CD). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0033]

[0060] It is an object of the present invention to provide SPM conjugates, their preparation and their use in pharmaceutical compositions, in which the SPM active moiety is stabilized against chemical degradation.

[0061] Despite their promising pharmacology, SPMs are chemically unstable in part due to the tendency of their conjugated double bond systems to isomerize to thermodynamically more favorable states, a tendency that is initially exhibited 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, docosahexaenoic, or docosapentaenoic acids, 20 or 22 carbon atoms long, with 5 or 6 double bonds arranged in diene, triene, and / or tetraene systems. For example, the 6Z,8E,10E triene system of resolvin E1 (RvE1) is thermodynamically unstable and consequently isomerizes to the lower energy all-trans structure. This isomerization results in the conversion of the 6,7-cis double bond to the 6,7-trans isomer, with significantly reduced biological activity, showing 70% less activity in various assays (Lombardo Thesis, Curtin University, January 2015, pp. 33-34).

[0034]

[0062] Structural modifications that address the inherent vulnerability of the conjugated double bond structure of SPMs, specifically the tendency of diene, triene, and tetraene systems to isomerize to less potent trans configurations, have been proposed to stabilize SPMs. Lombardo proposed structural modifications of RvE1 to stabilize the molecule, specifically through the formation of a benzene analogue that blocks isomerization of the 6,7-cis double bond. The use of cyclopropane and benzene congeners of Resolvin E2 (RvE2) has been proposed 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) through the insertion of a benzo-fused ring system that counteracts 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 Chem Lett. 2008;18(4):1382-138). Similarly, the use of a benzene-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 SPMs against isomerization, they may also result in reduced potency compared to the native unmodified SPM molecule. For example, Lombardo reports that benzo-resolvin E1 analogs have four-fold lower binding affinity to one of the known RvE1 receptors compared to the native RvE1 molecule (Lombardo, 2015, p. 113). Similarly, the benzo-lipoxin A4 compounds described by Petasis are less biologically active and have potencies in the range of 20%-50% lower compared to control lipoxin A4 analogs (Table 2, Petasis, 2008). In addition to potential loss of biological activity, structural SPM analogs may present additional safety and / or efficacy issues due to off-target effects compared to native unmodified SPM molecules.

[0035]

[0063] An alternative strategy is described in WO2017 / 210604, which shows that the formation of amino acid chelate salts of minerals of natural SPMs, such as magnesium dilysinate resolvin E1 (RvE1-MgLys), increased their solid-state chemical stability under standard conditions, i.e., 20-24°C, 40% relative humidity, and exposure to air. However, further testing by the inventors revealed that such compounds exhibited undesirable levels of chemical stability over time under accelerated conditions, i.e., 40°C, 75% relative humidity, and exposure to air. The present invention addresses the need to further stabilize SPMs against chemical instability when exposed to air under conditions of high temperature and humidity, which reduces the risk of loss of potency and / or appearance of unknown impurities along with potential safety issues during manufacture, formulation, packaging, storage, and distribution.

[0036]

[0064] The present invention is based in part on the unexpected discovery that complexation 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 due to exposure to light, air, and humidity. Similar stabilization against isomerization is expected for other SPMs with similar chemical structures, in particular conjugated double bond systems consisting of diene, triene, and / or tetraene systems, including the SPMs listed in Table 1 below.

[0037]

[0065] Thus, the present invention provides complexes of SPMs or their salts, or esters, or amides of SPM molecules, and cyclodextrins, where the SPMs contain diene, triene, and / or tetraene systems, as well as methods for making the complexes and for their use in therapy. These complexes are advantageously stabilized against chemical degradation, especially when exposed to air under conditions of high temperature and humidity (i.e., 40° C. and 75% relative humidity) for periods of up to 16 or 24 weeks, resulting in significant degradation of the SPM molecules and a concomitant decrease in chemical purity with an increase in degradation products that may both prevent the use of SPMs as pharmaceuticals. For example, after 16 weeks of exposure to air under the above-mentioned conditions of high temperature and humidity, the RvE1 moiety of RvE1-MgLys alone was significantly degraded from 94.0% to 80.7%, while the RvE1 moiety of five different cyclodextrins complexed with RvE1-MgLys was degraded on average from 95.4% to 91.7%. Thus, compared to RvE1-MgLys alone, cyclodextrin reduced RvE1 degradation by an average of 70%, and γ-cyclodextrin reduced RvE1 degradation by 91%. As used herein, the term "chemical purity" refers to the amount given as an area percentage of the active moiety of a particular compound in a sample of the compound. Unless otherwise stated, the percentages stated throughout this specification for chemical purity are area percentages calculated as the ratio of (a) the area of ​​the peak corresponding to the SPM active moiety, isomer, epimer, other impurity, or degradation product to (b) the sum of the peak areas of the detectable components of the sample measured by high pressure liquid chromatography (HPLC).

[0038]

[0066] Cyclodextrins are known to affect the chemical and photostability of various small molecules, including eicosanoids, but a comprehensive review by Popielec and Loftsson in 2017 also identified several molecules that are 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, testing of various cyclodextrins complexed with a prostaglandin E1 analog in solution (phosphate buffer, pH 8.0 at 60°C) showed that six of eight cyclodextrins tested promoted the degradation of the analog compared to the control (see Table 1, Uekama K et al., Pharm Res. 2001;18(11):1578-1585). Furthermore, molecules stabilized with 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 promote and slow down various types of reactions depending on the nature of the complexes formed". In particular, the degree of encapsulation and orientation of guest molecules in cyclodextrins plays an important role in determining the stabilizing effect of cyclodextrins, especially with respect to the reactive sites of the molecules (Popielec, 2017). Hence, "the use of cyclodextrins to improve the stability of a given drug is unpredictable and should be thoroughly tested in carefully designed formulations" (Rincon-Lopez, 2021).

[0039] Complex

[0067] The present invention relates to a complex of an SPM molecule, or a salt, or an ester, or an amide thereof, and a cyclodextrin.

[0040] Cyclodextrin component of the complex

[0068] The term "cyclodextrin" refers to a cyclic oligosaccharide consisting of at least six glucopyranose units joined by α-1,4 glycosidic bonds, where the oligosaccharide ring forms a toroid or cone-like structure. In the context of the present invention, cyclodextrins with six glucose subunits are referred to as α-cyclodextrins, cyclodextrins with seven glucose subunits are referred to as β-cyclodextrins, and cyclodextrins with eight glucose subunits are referred to as γ-cyclodextrins. Collectively, these cyclodextrins are known as natural cyclodextrins. In addition, the term "cyclodextrin" includes, but is 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, heptakis(2,3,6-tri-O-methyl)-β-cyclodextrin; carboxymethyl-β-cyclodextrin; carboxyethyl-β-cyclodextrin; hydroxyethyl- It also refers to randomly substituted derivatives of natural cyclodextrins, including, but not limited to, β-cyclodextrin; maltosyl-β-cyclodextrin; 3,6-(N,N,N-trimethylammonium)propyl-β-cyclodextrin; acetyl-β-cyclodextrin; sulfobutylether β-cyclodextrin; 2,6-di-O-methyl-γ-cyclodextrin; 2-hydroxypropyl-γ-cyclodextrin; and sulfobutylether γ-cyclodextrin.

[0041]

[0069] The term "complex" refers to an inclusion compound in which a "guest" molecule is partially or completely contained within the void spaces or cavities of a "host" molecule or within the lattice of the 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").

[0042] SPM components of the complex

[0070] The present invention refers to special pro-resolution mediators (SPMs), or their salts, or esters, or their amides, and cyclodextrin complexes, where the SPMs contain dienes, trienes, and / or tetraenes. SPMs include resolvins, protectins, lipoxins, and maresins, as well as their aspirin-induced counterparts (e.g., aspirin-induced lipoxins and protectins). These molecules are described, for example, in US5,441,951 and US8,119,691 (lipoxins and aspirin-induced lipoxins), US6,670,396 (aspirin-induced lipid mediators), US2006-0293288 (resolvins), US7,378,444, and US7,595,341 (analogs of lipid mediators derived from omega-3 fatty acids). The esters of SPMs can be isopropyl, methyl, ethyl, or glycerol esters. (Obrosov A et al., J Neurol Neurophysiol.2017;8(6)).

[0043]

[0071] In embodiments, the SPM component of the conjugates described herein is selected from the molecules described in Table 1. In embodiments, the SPM component of the conjugates described herein is a free acid or a salt, or an ester, or an amide thereof of an SPM described in Table 1. In embodiments, the SPM component of the conjugates described herein is selected from the molecules described in Table 1. In embodiments, the SPM component of the conjugates described herein is selected from the molecules described in Table 1. 1 , and MaR 2 , PD1 n-3 DPA Or MaR 1 n-3 DPA or a salt, ester, or amide thereof.

[0044] [Table 1-1]

[0045] [Table 1-2]

[0046] salt

[0072] The SPM component of the SPM-cyclodextrin complexes described herein can be a salt of an SPM molecule. In embodiments, the salt of an SPM molecule can be a pharma- ceutically acceptable base addition salt, including salts with basic organic or inorganic moieties; or a salt according to Formula I, or a dipeptide and polypeptide salt described in WO2017 / 210604 or WO2019 / 108605.

[0047] Base addition salts

[0073] In an embodiment, SPM salts, which consist of pharma- ceutically acceptable base addition salts, are prepared from the addition of inorganic or organic bases 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. These include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines, including naturally occurring substituted amines, cyclic amines, and basic 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, methylglucamine, 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.

[0048]

[0074] In an embodiment, the SPM salt contains two molecules of the same SPM as described in formula I, ionically bonded to two basic functional groups provided in a scaffold as described in formula I, the scaffold being a divalent metal-amino acid chelate or a divalent metal-peptide chelate. In general, the carboxylic acid moiety of the molecule forming the SPM molecule or SPM component is deprotonated to form an ionic bond with a basic functional group (or functional groups) of the scaffold portion of the compound. These scaffolds and their SPM salts are described in WO2017 / 210604 and WO2019 / 108605. SPMs that can form the SPM components of the salt of formula I according to the present invention are shown in Table 1. It is understood that the neutral compounds described in the table can be loaded (i.e., deprotonated) when solvated at the appropriate pH to form the anionic components described in the compound of formula I. The compound represented by formula I contains two SPM molecules (which may be referred to herein as the "SPM components" of the compound), and a scaffold portion to which the SPM components are ionically bonded, respectively. The term "bis" refers to two (bis)SPM molecules in a salt compound. In embodiments, the two SPM molecules are the same and include 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, MaR 1 , and MaR 2 , PD1 n-3 DPA , or MaR 1 n-3 DPA is selected from.

[0049]

[0075] Abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts. For example, when a substituent such as -NH3 is shown without a charge, the formal charge, i.e., NH3 + It is understood that the

[0050]

[0076] The term “alkyl,” by itself or as part of another substituent, means, unless otherwise stated, a straight-chain (i.e., unbranched) or branched-chain carbon chain (or carbons), or combinations thereof, which may be fully saturated, monounsaturated, or polyunsaturated, and which may include monovalent, divalent, and polyvalent radicals having the specified number of carbon atoms (i.e., C1-C6). 10 means 1 to 10 carbons).

[0051]

[0077] 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 embodiments, a basic functional group is -NH + , -NHC(NH2 + )NH2, -NHR 6 R 7 , -NR 6 R 7 R 8 R 6 , R 7 , and R 8 are each independently hydrogen, -CN, -COOH, -CONH, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl; R 6 and R 7 The substituents can be optionally linked to form an unsubstituted heterocycloalkyl or an unsubstituted heteroaryl. In an embodiment, the basic functional group is a hydrogen bond acceptor. In an embodiment, the basic functional group is a positively charged amine.

[0052]

[0078] It is understood that due to resonance, charge may be distributed throughout the molecule. The chemical structures and formulas set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts, and therefore those of ordinary skill in the art will recognize the equivalence of moieties having resonance structures.

[0053]

[0079] In embodiments, "amino acid side chain" or "side chain" as used herein is used according to its ordinary meaning and refers to a functional substituent contained in naturally occurring amino acids. 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, e.g., hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. In embodiments, the side chain of an amino acid is ionized (e.g., carries a formal charge).

[0054]

[0080] In an embodiment, the side chain is H,

[0055] [ka]

[0056] is selected from the group consisting of:

[0081] In an embodiment, the side chain is H. In an embodiment, the side chain is

[0057] [ka]

[0058] In an embodiment, the side chain is:

[0059] [ka]

[0060] In an embodiment, the side chain is:

[0061] [ka]

[0062] In an embodiment, the side chain is:

[0063] [ka]

[0064] In an embodiment, the side chain is:

[0065] [ka]

[0066] In an embodiment, the side chain is:

[0067] [ka]

[0068] In an embodiment, the side chain is:

[0069] [ka]

[0070] In an embodiment, the side chain is:

[0071] [ka]

[0072] In an embodiment, the side chain is:

[0073] [ka]

[0074] In an embodiment, the side chain is:

[0075] [ka]

[0076] In an embodiment, the side chain is:

[0077] [ka]

[0078] In an embodiment, the side chain is:

[0079] [ka]

[0080] It is.

[0082] In embodiments, the side chains can be optionally attached to adjacent nitrogens to form an unsubstituted heterocycloalkyl (eg, pyrrolidinyl).

[0081]

[0083] In an embodiment, the side chain is

[0082] [ka]

[0083] In an embodiment, the side chain is:

[0084] [ka]

[0085] In an embodiment, the side chain is:

[0086] [ka]

[0087] In an embodiment, the side chain is:

[0088] [ka]

[0089] In an embodiment, the side chain is:

[0090] [ka]

[0091] In an embodiment, the side chain is:

[0092] [ka]

[0093] In an embodiment, the side chain is:

[0094] [ka]

[0095] In an embodiment, the side chain is:

[0096] [ka]

[0097] It is. The side chain of glycine is H. The side chain of arginine is

[0098] [ka]

[0099] The side chain of histidine is

[0100] [ka]

[0101] The side chain of lysine is

[0102] [ka]

[0103] The side chain of aspartic acid is

[0104] [ka]

[0105] The side chain of glutamic acid is

[0106] [ka]

[0107] The side chain of serine is

[0108] [ka]

[0109] The side chain of threonine is

[0110] [ka]

[0111] The side chain of asparagine is

[0112] [ka]

[0113] The side chain of glutamine is

[0114] [ka]

[0115] The side chain of cysteine ​​is

[0116] [ka]

[0117] The side chain of proline is

[0118] [ka]

[0119] The side chain of alanine is

[0120] [ka]

[0121] The side chain of valine is

[0122] [ka]

[0123] The side chain of isoleucine is

[0124] [ka]

[0125] The side chain of leucine is

[0126] [ka]

[0127] The side chain of methionine is

[0128] [ka]

[0129] The side chain of phenylalanine is:

[0130] [ka]

[0131] The side chain of tyrosine is

[0132] [ka]

[0133] The side chain of tryptophan is

[0134] [ka]

[0135] It is. The term "unnatural 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 bound to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium, allylalanine, 2-aminoisobutryric acid. Unnatural amino acids are non-proteinogenic amino acids that are either naturally occurring or chemically synthesized. Such analogs have modified R groups (e.g., norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Non-limiting examples include exo-cis-3-aminobicyclo[2.2.1]hept-5-ene-2-carboxylic acid hydrochloride, cis-2-aminocycloheptane carboxylic acid hydrochloride, cis-6-amino-3-cyclohexene-1-carboxylic acid hydrochloride, cis-2-amino-2-methylcyclohexane carboxylic acid hydrochloride, cis-2-amino-2-methylcyclopentane carboxylic 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-piperazino)-2-(3,4-Dimethoxyphenyl)acetic acid purum, 2-(4-Boc-piperazino)-2-(2-fluorophenyl)acetic acid purum, 2-(4-Boc-piperazino)-2-(3-fluorophenyl)acetic acid purum, 2-(4-Boc-piperazino)-2-(4-fluorophenyl)acetic acid purum, 2-(4-Boc-piperazino)-2-(4-methoxyphenyl)acetic acid purum, 2-(4-Boc-piperazino)-2-phenylacetic acid purum, 2-(4-Boc-piperazino)-2-(3-pyridyl)acetic acid purum, 2-(4-Boc-piperazino)-2-[4-(trifluoromethyl)phenyl]acetic acid purum, Boc-β-(2-quinolyl)-Ala-OH, N-Boc-1,2,3,6-tetrahydro-2-pyridine carbamide Boc-β-(4-thiazolyl)-Ala-OH, Boc-β-(2-thienyl)-D-Ala-OH, Fmoc-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.

[0136] Compounds of Formula I In an aspect, the present disclosure provides a compound of formula I:

[0137] [ka]

[0138] The present invention provides a compound of the formula: [In the formula, M is a divalent metal; A 1 and A 2 are each the same SPM anion; R 1 and R 2 Each independently represents a C1 to C6 alkyl group having at least one basic functional group. 10 is alkyl; X 1 and X 2 are each independently H or CO-Z, and Z is a peptide comprising 1 to 5 amino acids or a pharma- ceutically acceptable salt thereof. The compound of formula I includes two amino acid moieties and two SPM molecules coordinated around a divalent metal cation as an amino acid component. In an embodiment, the divalent metal cation is Mg 2+ , Ca 2+ , Mn 2+ , Fe 2+ , Cu 2+ , Co 2+ , Ni 2+ , Mo 2+ , or Zn 2+ In an embodiment, the divalent metal cation is Mg 2+ In an embodiment, the divalent metal cation is Ca 2+ In an embodiment, the divalent metal cation is Zn 2+ It is.

[0139] In an embodiment, the amino acid component comprises lysine or arginine, or consists of lysine or arginine. In an embodiment, the amino acid component comprises lysine or arginine. In an embodiment, R 1 and R 2 The basic functional group is selected from primary amines, secondary amines, tertiary amines, and guanidines. In an embodiment, the basic functional group is -NH3, -NHC(NH2 + )NH2, -NHR 6 R 7 , or -NR 6 R 7 R 8 R 6 , R 7 , R 8are each independently hydrogen, -CN, -COOH, -CONH, unsubstituted alkyl, unsubstituted heteroalkyl, unsubstituted cycloalkyl, unsubstituted heterocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl; 6 and R 7 The substituents can be optionally joined to form an unsubstituted heterocycloalkyl or an 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.

[0140] In an embodiment, R 1 and R 2 are each a side chain of an amino acid residue having a basic functional group. 1 and R 2 are the same, and the amino acid residue is lysine or arginine.

[0141] In an embodiment, R 1 and R 2 is -(CH2)3-Y 1 , and -(CH2)4-Y 2 are independently selected from Y 1 and Y 2 are each a basic functional group, which may be the same or different. 1 is -CH2CH2NH3. In an embodiment, R 2 is -CH2CH2NH3. In an embodiment, R 1 is -CH2CH2CH2CH2NH3. In an embodiment, R 2 is -CH2CH2CH2CH2NH3.

[0142] In an embodiment, R 1 and R 2 Both are -(CH2)4-Y 2 and Y 2 -NH3 + It is. In the embodiment of the present invention, R 1 and R 2 Both are -(CH2)3-Y1 and Y 1 -NHC(NH2 + )NH2.

[0143] In an embodiment, R 1 -(CH2)3-Y 1 and Y 1 -NHC(NH2 + )NH2, and Y 2 -(CH2)4-Y 2 and Y 2 -NH3 + In an embodiment, R 1 -(CH2)4-Y 2 and Y 2 -NH3 + and R 2 -(CH2)3-Y 1 and Y 1 is NHC(NH2 + )NH2.

[0144] In one embodiment, X 1 and X 2 are the same and are hydrogen (H). 1 is hydrogen. 2 is hydrogen. In one embodiment of the compound of formula I, A 1 and A 2 RvE1, RvE2, RvE3, RvE4, AT-RvE1, AT-RvE2, AT-RvE3, RvD1, RvD2, RvD3, RvD4, RvD5, RvD6, AT-RvD1, AT-Rv D2, AT-RvD3, AT-RvD4, AT-RvD5, AT-RvD6, RvT1, RvT2, RvT3, RvT4, LxA4, LxB4, AT-LxA4, AT-LxB4, PD1, PDXX 1 , AT-PD1, MaR 1 , and MaR 2 , PD1 n-3 DPA , or MaR 1 n-3 DPA M is selected from the group consisting of Mg 2+ , Ca 2+ , or Zn 2+ and R1 and R 2 are both -(CH2)4-Y 2 and Y 2 is NH3 + ;X 1 and X 2 is H. R 1 , R 2 , and Y 2 This selection may be referred to herein as a metal "dilysinate," e.g., "magnesium dilysinate" or "Mg-dilysinate." In this embodiment, the peptide component consists of a lysine dipeptide.

[0145] In one embodiment of the compound of formula I, A 1 and A 2 are the same and are selected from the E series of resolvins. In an embodiment, the E series of resolvins are selected from RvE1, RvE2, RvE3, RvE4, AT-RvE1, AT-RvE2, and AT-RvE3, and M is Mg 2+ , Ca 2+ , or Zn 2+ and R 1 and R 2 are both -(CH2)4-Y 2 and Y 2 is NH3 + ;X 1 and X 2 is H. R 1 , R 2 , and Y 2 This selection may be referred to herein as a metal "dilysinate," e.g., "magnesium dilysinate" or "Mg-dilysinate."

[0146] In one embodiment of the compound of formula I, A 1 and A 2 are the same and are selected from the D-series resolvins. In embodiments, 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, and M is Mg 2+ , Ca2+ , or Zn 2+ and R 1 and R 2 are both -(CH2)4-Y 2 and Y 2 is NH3 + ;X 1 and X 2 is H. R 1 , R 2 , and Y 2 This selection may be referred to herein as a metal "dilysinate," e.g., "magnesium dilysinate" or "Mg-dilysinate."

[0147] In one embodiment of the compound of formula I, A 1 and A 2 are the same and are selected from LxA4 or AT-LxA4; M is Mg 2+ , Ca 2+ , or Zn 2+ and R 1 and R 2 are both -(CH2)4-Y 2 and Y 2 is NH3 + ;X 1 and X 2 is H. R 1 , R 2 , and Y 2 This selection may be referred to herein as a metal "dilysinate," e.g., "magnesium dilysinate" or "Mg-dilysinate."

[0148] physical properties The complexes described herein and compositions comprising the same have advantageous chemical and physical properties compared to free SPM and its salts, esters, or amides. For example, in embodiments, the SPM component of the complexes described herein can be stabilized against chemical degradation as a solid or in solution in deionized water compared to the corresponding free SPM or SPM salt, ester, or amide under accelerated conditions of 40° C., 75% relative humidity, and exposure to air. In embodiments, the complexes are stable against chemical degradation, including oxidative degradation. In embodiments, the complexes are stabilized against degradation induced by exposure to air, oxygen, and humidity, as evidenced by a relative loss of change in physical properties, such as fluidity, and / or chemical properties, as measured, for example, by spectroscopic techniques such as nuclear magnetic resonance (NMR) or high pressure liquid chromatography (HPLC). In embodiments, the increased stability is evidenced by a relative loss of chemical degradation at 8 weeks, or 16 weeks, or 24 weeks, compared to SPM in the form of a sodium salt or mineral amino acid chelate salt. In embodiments, the SPM component of the complexes described herein is stabilized against chemical degradation as evidenced by a significant reduction in degradation products at 8 weeks, or 16 weeks, or 24 weeks, compared to SPM in its salt or ester form.

[0149] In an embodiment, the complexes are physically solid, free-flowing materials 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, the complexes maintain their form as solid, free-flowing materials for up to 168 hours without visible change when exposed to air at temperatures between 18-22°C and 50%-75% relative humidity, and change from a brown / orange powder to a gel-like or oil-lie material within 24 hours, as compared to the corresponding free SPM or SPM salt, ester, or amide. In an embodiment, the complexes can be blended with standard pharmaceutical excipients such as fillers, glidants, disintegrants, and lubricants with flow properties suitable for gravity feeding in a rotary press. For example, in an embodiment, when the complex is blended with a standard mixture of pharmaceutical excipients in a ratio of approximately 1 to 4.3 (w / w) without further dry or wet milling (i.e., granulation), the resulting blend has a particle size distribution with flow properties suitable for compression by a gravity-fed rotary press. In an embodiment, core tablets with tablet hardness between 18 kilopounds (8.1 kg) and 30 kilopounds (13.6 kg) produced using the aforementioned blend of 1 to 4.3 and using a gravity-fed rotary press have friability and disintegration characteristics consistent with specifications believed to be suitable for immediate release oral dosage forms or enteric coating as delayed release oral dosage forms. In addition, the complexes and compositions of the present invention can be easily combined.

[0150] Pharmaceutical Compositions The present disclosure provides a pharmaceutical composition comprising an SPM, such as a resolvin, protectin, lipoxin, or maresin, or a salt, or an ester, or an amide thereof, complexed with a cyclodextrin. In an embodiment, the salt of the SPM can be a simple salt, such as a sodium, potassium, calcium, zinc, or magnesium salt, any other pharma- ceutically acceptable base addition salt, or a salt according to Formula I.

[0151] The pharmaceutical compositions described herein may be formulated for any suitable route of administration, e.g., oral, pulmonary, rectal, parenteral, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, inhalational, buccal, sublingual, intrapleural, intrathecal, intranasal, vaginal, etc. Dosage forms for topical or transdermal administration of the compounds of the present invention include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants.

[0152] In an embodiment, the pharmaceutical composition is formulated as a parenteral dosage form, e.g., 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 the subcutaneous route.

[0153]

[0104] In an embodiment, the pharmaceutical compositions are in the form of sterile aqueous solutions or dispersions suitable for administration by either direct injection or addition of sterile infusion fluids for intravenous infusion, and include a solvent or dispersion medium containing water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), or suitable mixtures thereof.

[0154]

[0105] According to any of these embodiments, the dosage form may be in the form of a clear aqueous solution, which may optionally be frozen or lyophilized, preferably a sterile lyophilized solid, contained in a container such as a prefilled syringe, vial or ampoule. In some embodiments, the container contains the lyophilized composition and is suitable for reconstitution with a specified amount of sterile water, aqueous buffer or other commonly used parenteral diluent, such as 5% dextrose solution, saline solution, etc., for administration by parenteral routes, e.g., intravenous, intraarterial, or intramuscular, subcutaneous.

[0155]

[0106] In an embodiment, the pharmaceutical composition is formulated as an oral (or peroral) dosage form. In an embodiment, the oral dosage form is in the form of, for example, a tablet, capsule, powder, solution, suspension, or emulsion.

[0156] In an embodiment, the pharmaceutical composition is formulated as a sublingual dosage form. In an embodiment, the sublingual dosage form is in the form of a tablet, film or spray. In an embodiment, the pharmaceutical composition is formulated for administration via inhalation through the nose or mouth. In an embodiment, the inhalable dosage form is a liquid formulation, such as an aqueous solution formulation, adapted for pulmonary delivery via a nebulizer, including jet, vibrating mesh, and static mesh or orifice nebulizers. In an embodiment, the inhalable dosage form is a dry powder for inhalation (DPI). In an embodiment, the inhalable dosage form is a propellant-based aerosol formulation suitable for administration using a metered dose inhaler (MDI).

[0157] In an embodiment, the pharmaceutical composition comprising the SPM complex is in the form of a unit dose of the SPM in the complex. In an embodiment, the unit dosage form is a sterile, lyophilized composition in a suitable container, such as an ampoule or vial. The term "freeze-dried" is synonymous with the term "lyophilized" in this context. In an embodiment, the unit dose contains 0.5 μg to 100 mg of the SPM active moiety. In an embodiment, the unit dose contains 1, 25, 50, 100, 250, or 500 μg of the SPM active moiety. In an embodiment, the unit dose contains 1, 2, 3, or 5 mg, or multiples thereof between 1 and 100 mg, of the SPM active moiety.

[0158]

[0110] The compositions described herein may be formulated using one or more suitable excipients or carriers. Suitable excipients or carriers are those suitable for human or animal use. The term "excipient" refers to an additive that serves some purpose in the composition other than the carrier, for example, as a stabilizer, solubilizer, bulking agent, compression aid, flow improver, lubricant, disintegrant, or suspending agent. In many cases, a carrier serves a dual purpose as a simple carrier or diluent and excipient. Thus, examples of pharma- ceutical acceptable excipients may include carriers. Non-limiting examples of excipients for use in the compositions of the present invention include sterile liquids, water, buffered saline, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), oils, surfactants, 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.

[0159]

[0111] A suitable excipient or carrier is typically a pharma- ceutically acceptable carrier or excipient for use in humans or animals. The term "pharmaceutically acceptable" refers to approval by a federal or state regulatory agency, or approval by a regulatory agency listed in the United States Pharmacopoeia, or other commonly recognized pharmacopoeias, such as the European Pharmacopoeia, for use in animals, more specifically humans. In the context of the pharmaceutical compositions of the present invention, "carrier" refers to, for example, a solvent, diluent, or vehicle in which the conjugate of the present invention is formulated for delivery. Examples of pharma- ceutically acceptable carriers for use in 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 (e.g., glucose, lactose, sucrose, or dextran) for solid dosage forms.

[0160]

[0112] The present disclosure also provides packaging and kits comprising pharmaceutical compositions for use in the methods of the present invention. The kits may comprise one or more containers selected from the group consisting of bottles, vials, ampoules, blister packs, infusion pumps, and syringes. The kits may further comprise one or more instructions for use, one or more syringes, one or more applicators, or a sterile solution suitable for reconstituting the pharmaceutical compositions of the present invention.

[0161] In an embodiment, the pharmaceutical composition is a solid dosage form comprising the SPM complex blended with one or more excipients or carriers. The blend of SPM complex and excipients can be filled directly into capsules or compressed into tablets, with or without prior granulation of the blend. In an embodiment, the SPM complex and excipients are present in a weight to 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.

[0162] Method for preparing the complex

[0114] The present invention provides a method for complexation of a cyclodextrin with an SPM, or a salt thereof, comprising dissolving the SPM, or a salt thereof, and the cyclodextrin in an aqueous solution, which may 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. The resulting complex can be purified by removal of 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 drying techniques, such as co-evaporation under reduced pressure, 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., J Pharm 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 July;35(7):796-807; Promzeleva M. et al., ACS Biomater. Sci. Eng. 2018;4:491-501).

[0163] Medicinal Use

[0115] The human inflammatory response has two phases: initiation and resolution. At the cellular level, initiation is a protective response characterized by the production and release of proinflammatory mediators that activate the innate immune response, resulting in the influx of polymorphonuclear cells (PMNs) from the cellular compartment that kills and clears foreign invaders and damaged cells. This initiation phase is an active process driven by metabolites of arachidonic acid, including prostaglandins, which are chemoattractants for eosinophils, neutrophils, and monocytes, and leukotrienes, which induce leukocyte adhesion, chemotaxis, and aggregation.

[0164] During peak inflammation, as inflammatory stimuli are reduced, the resolution phase begins, characterized by a reduction in proinflammatory signals and a switch from the production and release of proinflammatory mediators to the production and release of SPMs, including resolvins, protectins, lipoxins, and maresins. SPMs can activate and coordinate the resolution phase by halting PMN infiltration and stimulating the recruitment and activation of monocytes and pro-resolution macrophages to clear tissues of cellular debris in a non-inflammatory manner, restoring tissue homeostasis (Serhan et al., Cold Spring Harb Perspect Biol 2015;7:a016311).

[0165] Unresolved inflammation is widely recognized as a unifying aspect of many chronic diseases and disorders. Therefore, the ability to resolve excessive inflammation is of paramount importance 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).

[0166]

[0118] Researchers have demonstrated the efficacy of SPM in preventing and / or treating disease in many preclinical models, including those related to Alzheimer's disease, burn wounds, chronic pancreatitis, diabetic wounds, dermatitis, pulmonary inflammation, peripheral nerve injury, obesity, allergic airway response, amyotrophic lateral sclerosis, acute lung injury, fibrosis, bacterial infection, peritonitis, dry eye, tissue regeneration, pain, adipose tissue inflammation, localized active periodontitis, colitis, temporomandibular joint inflammation, arthritis, postoperative pain, postoperative cognitive decline, endotoxic shock, HSV-keratitis, allograft rejection, cardiac ischemia, bacterial pneumonia, cigarette smoke-induced pulmonary inflammation, vascular inflammation, fibromyalgia, and vagotomy (Serhan CN et al., Cold Spring Harb Perspect Biol 2015;7:a016311).

[0167]

[0119] Lim et al. describe the analgesic effects of SPMs in multiple inflammatory pain models and characterize resolvins and related substances as therapeutic candidates to prevent the worsening of inflammation and pathological pain (Lim JY et al., BioMed Research International, 2015, pp. 1-14). Lim also notes that the "strong potency" and "negligible adverse effects" of these molecules make them attractive candidates for clinical use.

[0168]

[0120] U.S. Patent Nos. 8,008,282 and 6,627,658 describe lipoxin analogs and their use as inhibitors of angiogenesis. 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 to treat cell proliferation disorders. US Pat. No. 8,119,691 describes lipoxins and aspirin-triggered lipoxins and their analogs in the treatment of asthma and inflammatory airway diseases.

[0169]

[0121] US 2006-0293288 describes the use of resolvins to treat 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, colonic polyps, familial polyposis syndrome, Gardner's syndrome, Helicobacter pylori, irritable bowel syndrome, nonspecific diarrhea, 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 disclose resolvins and their anti-inflammatory, vascular proliferation, Its use in methods for treating cardiovascular, thrombophlebotic, vascular, ophthalmic, dermatological, neurodegenerative, pulmonary, endocrine, reproductive, rheumatic and gastrointestinal diseases, as well as methods for treating cardiovascular diseases, for example inflammation, including Crohn's disease, ulcerative colitis, distal proctitis, rheumatic spondylitis, arthritis, rheumatoid arthritis, osteoarthritis, gouty arthritis, psoriasis, skin diseases, and the like, is described. dermatitis, eczematous dermatitis, atopic or seborrheic dermatitis, allergic or irritant contact dermatitis, eczema craquelee, photoallergic dermatitis, phototoxic dermatitis, phytophotodermatitis, radiation dermatitis, stasis dermatitis, arterial inflammation, coronary infarction injury, restenosis, uveitis, iritis, conjunctivitis, adult respiratory distress syndrome, bronchitis, cystic fibrosis, contractile irritant conditions, asthma, idiopathic bronchial asthma, contraction of arterial smooth muscle, coronary artery spasm, myocardial infarction, ischemia-induced myocardial injury, cerebral spasm, stroke, inflammatory bowel disorder, 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.

[0170]

[0122] US2012-0245229 describes resolvins and methods of treating neuropathic pain, including pain associated with diabetic neuropathy or HIV infection, post-operative pain, inflammatory pain, pain associated with cancer, and pain associated with fibromyalgia by administering resolvins.

[0171] No. 10,233,167 discloses maresin and its derivatives, which are useful in treating inflammation associated with neurodegeneration, memory loss, wrinkles, psoriasis, pityriasis capitis, dermatitis, arterial inflammation, arthritis, psoriasis, urticaria, vasculitis, asthma, eye inflammation, lung inflammation, pulmonary fibrosis, seborrheic dermatitis, pustular dermatitis, AIDS, allergic responses, Alzheimer's disease, inflammatory diseases, atherosclerosis, bone diseases, breast cancer, cancer, cardiovascular disease, colon cancer, birth defects, degenerative neurological diseases, dementia, skin disorders, diabetes, endocrinology, Methods for treating or preventing urinary disorders, eye disorders, gastrointestinal disorders, genitourinary disorders, hearing disorders, hematological disorders, hepatobiliary disorders, hypertension, immunological disorders, infectious diseases, leukemia / lymphoma, lung cancer, metabolic disorders, neonatology, neurological disorders, neuromuscular disorders, obesity / eating disorders, orthopedic 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 are described.

[0172]

[0124] US2020-0010398 and US2020-0048177 disclose novel monohydroxy, dihydroxy and trihydroxy docosapentaenoic acid derivatives and their applications in inflammation, arthritis, psoriasis, urticara, vasculitis, asthma, eye inflammation, lung inflammation, pulmonary fibrosis, seborrheic dermatitis, pustular dermatosis, cardiovascular disease, neutrophil, leukocyte and / or cytokine recruitment, addiction, AIDS, alcohol-related disorders, allergies, Alzheimer's disease, arthritis, asthma, atherosclerosis, bone disease, breast cancer, cancer, cardiovascular disease, colon cancer, birth defects, decision analysis , degenerative neurological disorders, dementia, dermatology, diabetes, endocrine disorders, eye disorders, fetal and maternal medicine, gastrointestinal disorders, gene therapy, genetic diagnosis, genetics, genitourinary disorders, geriatrics, growth and development, hearing, hematological 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, wounds, oral infections, periodontal disease, brain injury, trauma, and neuronal inflammation are described.

[0173]

[0125] 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, regulating blood glucose, inhibiting lipid-induced inflammation and other related inflammatory conditions, such as insulin resistance, metabolic syndrome, type 2 diabetes, hypertension and cardiovascular disease.

[0174] In an aspect, the invention provides a method of treating a disease or disorder modifiable by treatment with an SPM, including cancer, gastrointestinal diseases and disorders, inflammatory disorders, infectious diseases and disorders, metabolic diseases and disorders, neurological disorders, and pulmonary diseases and disorders. In an aspect, the method comprises administering to a subject in need of such treatment a pharmaceutical composition containing a complex of an SPM and a cyclodextrin as described herein, the complex comprising an amount of the SPM effective to treat the disease or disorder. With respect to the methods described herein, the term "treating" refers to regression, stabilization, or reduced progression of the disease or disorder being treated, or improvement or stabilization of one or more clinical symptoms associated with the disease or disorder being treated.

[0175] cancer

[0127] The present disclosure provides a method for treating cancer in a subject in need of such treatment, comprising the step of administering to the subject a pharmaceutical composition containing a complex described herein, wherein the complex contains an amount of SPM effective to treat the cancer.

[0176]

[0128] According to the methods described herein, administration can be alone as a monotherapy or in combination with one or more additional therapeutic agents. In aspects, the combination therapy is an adjuvant therapy, for example, as an adjuvant to chemotherapy, targeted therapy, immunotherapy, hormonal therapy, gene therapy, or microbiome therapy. In aspects, the combination therapy is an immune checkpoint inhibitor therapy. In aspects, the combination therapy is an adjuvant or neoadjuvant therapy to surgery or radiation therapy. In aspects, the composition can be administered as a first-line, second-line, third-line, or symptomatic treatment, alone or in combination with other therapies, for example, surgery, radiation therapy, chemotherapy, targeted therapy, immunotherapy, hormonal therapy, gene therapy, or microbiome therapy. The term "therapy(s)" refers to any method, protocol, and / or agent that can be used in the treatment or management of cancer.

[0177]

[0129] With respect to the methods described herein, the term "treating" can refer to the improvement or stabilization of one or more clinical symptoms associated with the cancer being treated. In an embodiment, treating results in the elimination of the clinical symptoms of the cancer being treated, but elimination is not necessary. In an embodiment, the severity of the symptoms is reduced. With respect to cancer, such symptoms can include clinical markers of severity or progression, including the size of a solid tumor, the amount of growth factors or other biomarkers secreted by a solid tumor, tumor vascularization, tumor metastasis, or number of metastases. Thus, treating cancer with the methods described herein can result in a reduction in tumor size or volume (also referred to as tumor regression); a reduction in tumor number; or a reduction in the number of metastatic lesions in other tissues or organs distant from the primary tumor site. Such symptoms can also include cancer cachexia or pain. Thus, treating cancer with the methods described herein can result in a reduction in cancer cachexia or pain, and the reduction in cancer cachexia or pain is due to administration of a complex as described herein. In embodiments, "treating" cancer by the methods described herein results in an increase in overall survival and / or progression-free survival in the treated subject. In the context of this disclosure, the term "treating" is not meant to encompass, unless expressly stated, any side effects or adverse events related to another therapy, such as radiation therapy or chemotherapy, that are not related to the cancer being treated, but instead, for example, any improvement or stabilization of clinical symptoms associated with the treatment modality. In addition, the term "treating" is not intended to encompass pre-diagnosis prevention of cancer in the treated subject or administration of an SPM complex.

[0178] In an aspect, the present invention relates to a method for treating adverse events or side effects of chemotherapy, targeted therapy, immunotherapy, or radiation therapy, such as gastrointestinal toxicity, e.g., oral or gastrointestinal mucositis, colitis, enteritis, gastritis, nausea, vomiting, and diarrhea; cardiovascular toxicity, e.g., cardiomyopathy, myocardial ischemia, pericarditis, myocarditis, valvular heart disease, arrhythmias, congestive heart failure, hypertension, coronary, cerebral, and peripheral vascular events, or congestive heart failure; dermatological toxicity, e.g., pruritus or severe rash; hepatotoxicity; pulmonary toxicity, e.g., pulmonary hypertension and pneumonia; endocrine disorders, e.g., hypothyroidism, hyperthyroidism, and hypophysitis; neurotoxicity, e.g., Guillain-Barré syndrome, severe In one embodiment, a method is provided for the amelioration of myasthenia gravis, reversible posterior leukoencephalopathy syndrome, aseptic meningitis, enteric neuropathy, transverse myelitis, pancerebellitis, autoimmune encephalitis, and cerebral and peripheral neuropathies, or other side effects such as pancreatitis, acute kidney injury, and cytokine release syndrome, comprising administering a conjugate as described herein to a subject undergoing or likely to undergo chemotherapy, targeted therapy, immunotherapy, or radiation therapy, wherein the resolvin is effective to alleviate or prevent adverse events or side effects of chemotherapy, targeted therapy, immunotherapy, or radiation therapy.

[0179]

[0131] According to the methods described herein, a therapeutically effective amount of an SPM conjugate is administered to a subject in need of treatment. A therapeutically effective amount is an amount or dose of an SPM active moiety sufficient to treat cancer or to achieve a desired therapeutic outcome, e.g., improvement or stabilization of one or more biomarkers or one or more clinical symptoms of disease progression.

[0180] Gastrointestinal Diseases and Disorders

[0132] In an aspect, the present disclosure provides a method for treating a gastrointestinal disease or disorder in a subject in need thereof by administering to the subject a pharmaceutical composition containing a conjugate described herein, wherein the conjugate contains an amount of SPM sufficient to treat the gastrointestinal disease or disorder.

[0181] In an embodiment, 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 an embodiment, the gastrointestinal disease or disorder is selected from ulcerative colitis, and Crohn's disease.

[0182] In an embodiment, the gastrointestinal disease or disorder is selected from intestinal obstruction, chronic pancreatitis, colitis, colon cancer, congenital gastrointestinal dysplasia, gastroschisis, high-output fistula, parenteral nutrition-associated liver disease, postoperative ileus, postoperative intestinal inflammation, short bowel syndrome, and sporadic polyposis. In an embodiment, the gastrointestinal disease or disorder is selected from eosinophilic esophagitis, Behcet's disease, irritable bowel syndrome, celiac disease, intestinal mucositis, NSAID enteropathy, intestinal infection, diverticulosis, diverticulitis, gastritis, pancreatitis, viral gastroenteritis, and Whipple's disease.

[0183] In an embodiment, the gastrointestinal disease or disorder is post-operative inflammation of the intestine, post-operative ileus, or a combination thereof. In an embodiment, the gastrointestinal inflammatory disease or disorder is post-operative ileus (POI). In a preferred embodiment, the gastrointestinal disease or disorder is inflammatory bowel disease (referred to herein as "IBD"), which is the 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 often affects the lower gastrointestinal tract, particularly the ileum and / or colon. Ulcerative colitis affects the colon and rectum. The terms "large intestine" and "colon" are used interchangeably in this disclosure.

[0184] Standard 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-aminosalicylates (5-ASA) can be used alone or in combination with corticosteroids to induce remission. Generally, corticosteroid treatment is administered for short-term control of symptoms, e.g., to treat acute flare-ups and induce remission of disease symptoms. Long-term or maintenance therapy generally involves the use of nonsteroidal agents, including 5-ASA, and immune modulators, such as azathioprine.

[0185] Standard 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 using adalimumab, golimumab, or infliximab, alone or in combination with corticosteroids, thiopurines, or 5-ASA, 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 involves the continuation of anti-TNF therapy with or without thiopurines, or other therapies as mentioned above.

[0186] In an aspect, the disclosure provides for the treatment of IBD by administering to a subject a pharmaceutical composition containing a conjugate as described herein, either as a monotherapy or in combination with a second IBD therapeutic. In an aspect, the second IBD therapeutic is selected from a corticosteroid, 5-ASA, azathioprine, an anti-TNF therapy, an integrin receptor antagonist, a sphingosine-1-phosphate receptor modulator, or a Janus kinase inhibitor. In an aspect, the disclosure provides bisRvE1 magnesium dilysinate for use in an IBD maintenance therapy, optionally in combination with 5-ASA, azathioprine, an anti-TNF therapy, an integrin receptor antagonist, a sphingosine-1-phosphate receptor modulator, or a Janus kinase inhibitor, for use in a method of treating IBD. In an embodiment, the present disclosure provides bisRvE1 magnesium dilysinate for use in a method for treating IBD in combination with 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 bisRvE1 magnesium dilysinate for use in a method for treating IBD in combination with corticosteroids. According to the above embodiment of the combination therapy of bisRvE1 magnesium dilysinate with corticosteroids, the corticosteroid can be selected from the group consisting of prednisone, prednisolone, budesonide, hydrocortisone, and beclomethasone dipropionate (the dosage form is an oral dosage form), or from hydrocortisone, prednisolone, and budesonide (the dosage form is adapted for rectal delivery, for example, in the form of an enema or suppository). According to the above-mentioned aspects of bisRvE1 magnesium dilysinate combination therapy with anti-TNF therapy, integrin receptor antagonist, sphingosine-1-phosphate receptor modulator, or Janus kinase inhibitor, the second IBD therapeutic agent can be selected from the group consisting of adalimumab, golimumab, infliximab, vedolizumab, ozanimod, etrasimod, tofacitinib, or filgotinib.

[0187] Inflammatory disorders

[0140] The compounds described herein may be particularly useful in treating diseases and disorders that have a significant inflammatory component due to the ability of SPMs to mediate the resolution of inflammation and the ability of the compounds described herein to deliver therapeutically effective amounts of SPMs to tissues of a subject in need of treatment for inflammation.

[0188]

[0141] In an aspect, the present disclosure provides a method for treating an inflammatory disease or disorder in a subject with a pharmaceutical composition containing a complex described herein, wherein the complex contains an amount of SPM sufficient to treat the inflammatory disease or disorder.

[0189] In an embodiment, the inflammatory disease or disorder is selected from the group consisting of acne, inflammation of adipose tissue, 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, localized active 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, Sjogren's syndrome, steroid-induced rosacea, stomatitis, systemic inflammatory response syndrome, temporomandibular joint inflammation, and vascular inflammation.

[0190] Infections and disorders caused by infectious agents

[0143] In an aspect, the present disclosure provides a method for treating a disease or disorder caused by an infectious agent, such as a bacterium, fungus, or virus, in a subject in need thereof by administering to the subject a pharmaceutical composition containing a complex described herein, wherein the complex contains an amount of SPM sufficient to treat the infection or disorder.

[0191] In an embodiment, the disease or disorder is a bacterial infection. In an embodiment, the bacterial infection is bacterial pneumonia. In an embodiment, the bacterial infection is an E. coli infection. In an embodiment, the bacterial infection is a Mycobacterium tuberculosis infection.

[0192]

[0145] In an embodiment, the disease or disorder is a yeast infection. In an embodiment, the yeast infection is a Candida yeast infection.

[0146] In an embodiment, the disease or disorder is sepsis. In an embodiment, the sepsis is burn wound sepsis.

[0193] Metabolic Diseases and Disorders

[0147] In an aspect, the present disclosure provides a method for treating a metabolic disease or disorder in a human subject in need thereof by administering to the subject a pharmaceutical composition containing a conjugate described herein, wherein the conjugate comprises an amount of SPM sufficient to treat the metabolic disease or disorder.

[0194]

[0148] In an embodiment, the metabolic disease or disorder is diabetes, including type 2 diabetes or prediabetes, abnormal glucose metabolism manifesting as insulin resistance, hypertriglyceridemia, i.e., hypertriglyceridemia, mixed dyslipidemia, hypercholesterolemia, abnormal lipid metabolism manifesting as fatty liver, and combined abnormal glucose and lipid metabolism manifesting as obesity; or a dyslipidemia disorder selected from hypertriglyceridemia, hypercholesterolemia, and mixed dyslipidemia.

[0195]

[0149] 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.

[0196]

[0150] In accordance with the methods described herein, administration can be 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 embodiments, the antihyperlipidemic agent is selected from statins, cholesterol absorption inhibitors, CETP inhibitors, and pharma- ceutically acceptable salts and prodrugs of any of the foregoing.

[0197] Neurological disorders

[0151] In an aspect, the present disclosure provides a method for 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 contains an amount of SPM sufficient to treat the neurological disorder.

[0198] In embodiments, neurological diseases and disorders that can 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 disorder is selected from 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 injuries, or trauma. With respect to the methods described herein, the term "treating" can refer to the improvement or stabilization of one or more clinical symptoms associated with the disease or disorder being treated.

[0199] In an embodiment, the amount is effective to treat one or more symptoms of a neurological disorder. In an embodiment, the neurological disorder is a psychiatric disorder. In an embodiment, the psychiatric disorder is selected from attention deficit hyperactivity disorder (ADHD) and depression. In an embodiment, the neurological disease or disorder is post-operative cognitive dysfunction (POCD) or post-operative delirium.

[0200]

[0155] The present disclosure also provides a method for treating or managing pain. In an aspect, 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.

[0201]

[0156] In an aspect, 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.

[0202]

[0157] In aspects, the present disclosure also provides methods for treating or managing acute post-surgical pain and chronic back pain. Pulmonary Diseases and Disorders

[0158] In an aspect, the present disclosure provides a method for treating a pulmonary disorder in a human subject in need thereof by administering to the subject a pharmaceutical composition containing a conjugate described herein, wherein the conjugate comprises an amount of SPM sufficient to treat the pulmonary disease or disorder.

[0203]

[0159] In an embodiment, pulmonary and vascular diseases and disorders that can be treated include, but are not limited to, asthma, pulmonary inflammation, bronchopulmonary dysplasia (also known as chronic lung disease of infancy), cystic fibrosis, allergic airway response, acute lung injury, acute respiratory distress syndrome, lung injury, idiopathic pulmonary fibrosis, bacterial pneumonia, tobacco smoke-induced pulmonary inflammation, and vascular inflammation. EXAMPLES

[0204]

[0160] Despite their promising pharmacology in treating inflammatory diseases and cancer, SPMs are chemically unstable, in part due to the tendency of their cis double bonds to isomerize to the thermodynamically more favorable trans configuration, and have been found to have a lower potency than native SPMs. Therefore, the object of the present invention is to provide SPM complexes that allow their use in pharmaceutical compositions, in which the stability of the SPM active moiety is significantly improved.

[0205]

[0161] To improve the solid-state stability of resolvin E1 (RvE1), a resolvin that represents an SPM containing conjugated double bonds that are vulnerable to isomerization, we evaluated several approaches to protect RvE1 from exposure to oxygen, light, and moisture without using analogous modifications to the chemical structure of the SPM molecule. These approaches include: (i) suspension of RvE1-MgLys in various lipid excipients including pesceol, maisine, Labrasol ALF, Span 80, poloxamer Kollisov P124 and Tween 80; (ii) spray coating of RvE1-MgLys mixed with hydroxypropylcellulose in a 1:4 ratio (w / w) in isopropyl alcohol using the Wurster fluid bed process onto microcrystalline cellulose beads containing a seal coat of hydroxypropylcellulose; (iii) addition of antioxidants (0.5 w / w%) including α-tocopherol, ascorbyl palmitate, propyl gallate, BHT, and BHA in methanol to RvE1-MgLys and lyophilization to form a dry powder; and (iv) use of cyclodextrins including α-CD, β-CD, γ-CD, HP-β-CD and SBE-β-CD to form a complex with RvE1-MgLys. Importantly, only the complexation of RvE1-MgLys with cyclodextrin provided adequate stabilization of the RvE1 active portion. This finding is surprising because it indicates that the stabilizing effect of cyclodextrin can be positive or negative, depending on the specific interaction of the reactive portion of the guest molecule inside the cyclodextrin cavity, regardless of 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, a person skilled in the art would not be able to predict a priori that they would be stabilized by one or more cyclodextrins under the specific conditions tested as described below.

[0206] Experimentally, RvE1-MgLys was complexed with various cyclodextrins in solution, followed by lyophilization to form powders, and the resulting compounds were characterized by NMR spectroscopy studies to confirm their novelty throughout the complexation, the strength of the intramolecular interactions, and the unique orientation of RvE1 inside the CD cavity. The usefulness of these new compounds was established by their improved chemical stability over RvE1-MgLys alone over 16 weeks under accelerated conditions.

[0207] Preparation of complexes Example 1: General method for preparation of conjugates

[0163] The main techniques available for cyclodextrin complexation with water-soluble drugs include dissolving the guest and cyclodextrin in water or an organic solvent / water mixture, then isolating the complex by removal of the solvent via crystallization, coevaporation under reduced pressure, spray drying, or lyophilization. Based on the high solubility of RvE1-MgLys and the small quantities prepared in this experiment, the lyophilization approach was chosen for the initial preparation of the complex. However, the other isolation techniques mentioned above may be suitable for generating SPM complexes.

[0208] The following steps were used to prepare the SPM complexes for NMR and stability testing. First, a calculated amount of RvE1-MgLys was mixed with a calculated amount of various cyclodextrins in a 1:2 molar ratio (see Table 2), dissolved in 1400 μL of DO, stirred for 1 h, and filtered (0.22 μm, MF-Millipore). For NMR testing, the filtrate was transferred to a standard 5 mm glass NMR test tube for testing. For stability testing, the filtrate was lyophilized (VirTis BenchTop Pro freeze dryer) and transferred to an open tube.

[0209] [Table 2]

[0210] NMR interaction studies confirming complexation with cyclodextrin

[0165] 1H-NMR and 2D ROESY NMR experiments were performed on a 600 MHz Varian DDR NMR spectrometer equipped with a 5 mm inverse detection gradient (IDPFG) probe at 298 K. Standard pulse sequences and processing routines available in VNMRJ4.1 were used. 1H chemical shifts (δ) were referenced to the residual HOD peak (δ = 4.7900 ppm) present in DO.

[0211]

[0166] 1H spectra were recorded from 16 scans with a relaxation time of 2 seconds applied. 2D Rotating Frame Overhauser Enhancement Spectroscopy (ROESY) spectra were recorded from 8 scans / increment using a relaxation time of 2 seconds and 200 increments for the native cyclodextrin complex and 512 increments for the randomly substituted cyclodextrin complex, in each case the spin lock time was set to 300 milliseconds. Further analysis was performed using Heteronuclear Single Quantum Coherence (HSQC) NMR to determine the proton-carbon single bond correlations. A summary of the atomic numbering of RvE1-MgLys and α-CD, β-CD, γ-CD, HP-β-CD and SBE-β-CD cyclodextrins used for NMR spectroscopy studies is shown in Figures 1-3.

[0212] Example 2: NMR characterization of RvE1-MgLys alone and intramolecular interactions

[0167] RvE1-MgLys including complete assignment 1 The H-NMR spectrum is shown in Figure 4. The signals of RvE1 are indicated by Roman numerals, Greek letters are assigned to the signals of magnesium lysinate, and possible impurities for RvE1-MgLys are indicated by asterisks.

[0213]

[0168] As shown in FIG. 1From the H-NMR spectrum, three well-defined regions of RvE1 were identified: (1) the unsaturated region between δ1H=6.5-5 ppm; (2) the chiral center (containing a secondary hydroxyl group between δ1H=4.7-4 ppm); and (3) the aliphatic region between δ1H=2.5-0.7 ppm. The assignment of RvE1-MgLys was made using further 2D ROESY and 13C NMR experiments.

[0214]

[0169] To map the intramolecular interactions and any intermolecular interactions between the components of RvE1-MgLys, 2D ROESY analysis was performed (Figure 5). Example 3: NMR characterization and molecular interaction studies of RvE1-MgLys complexed with α-cyclodextrin As shown in Figure 6, the NMR signals of the α-CD complex are found in a well-defined region of the spectrum. The anomeric protons appear between δ1H=5.0-5.2 ppm, while the other protons of the glucopyranose units resonate above δ=3.5-4.0 ppm (frequently referred to as the core region).

[0215] Generally, complex formation occurs when a guest molecule and a host molecule 1 It induces a shift in the H-NMR spectrum so that previously non-overlapping signals between the guest and host become overlapping (called a multiplet), and the overlapping signals can be shifted partially or completely apart.

[0216]

[0172] The signal shift associated with the complex formation of RvE1-MgLys and α-CD was similar to that of the unsaturated region of RvE1-MgLys alone. 1 The H-NMR spectrum (Figure 7a) shows the unsaturated region of RvE1-MgLys in the α-CD complex. 1This was shown by comparing with the H-NMR spectrum (Figure 7b). Some signals in the unsaturated region of RvE1-MgLys that were previously separate became overlapping multiplets (e.g., 8 / 16 and 11 / 17), while some originally overlapping signals were shifted apart (7 / 15). Importantly, minimal shifts were observed in the saturated region of RvE1-MgLys alone compared to RvE1-MgLys in α-CD complex (Figure 8), indicating minimal interactions in this region of the RvE1 molecule.

[0217] 2D ROESY spectrum analysis was performed to provide detailed information on the plausible host-guest interactions of RvE1-MgLys with α-CD (FIG. 9). Since the signal of α-CD appears in a well-defined region of the spectrum, the likely regions of intermolecular interactions can be determined as shown by the dashed rectangle, whereas the specific interactions between the guest and the host are highlighted by the solid rectangle. Using 2D ROESY spectrum analysis, intermolecular correlations were shown between the protons of α-CD present inside the α-CD cavity (3' and 5') and on the main rim of the cavity (6') and the protons of RvE1-MgLys bound to the unsaturated carbons (6-12) and its adjacent carbon (5). Furthermore, no cross peaks were detected between the amino acid chelate (MgLys) component of RvE1-MgLys and α-CD.

[0218]

[0174] Figure 10 shows a close-up view of the intermolecular interactions, allowing the determination of the exact mode of interaction. These spectra show that the protons of RvE1-MgLys that interact with α-CD are 5, 7, 8, 9, 10 and 11. Based on these data, a reasonable orientation of RvE1 inside the α-CD cavity is shown in Figure 23a.

[0219] Example 4: NMR characterization and molecular interaction studies of RvE1-MgLys complexed with β-cyclodextrin

[0175] Figure 11 shows that the NMR signals of the β-CD complex are seen in a well-defined region of the spectrum similar to the α-CD complex.

[0220] As shown in Figure 7, the 8 / 16 and 6 / 14 multiplet signals were fully and partially resolved, respectively, while the 7 / 15 resolution was slightly reduced compared to RvE1-MgLys alone, indicative of complex formation. The smallest signal shift was observed at the saturation portion of RvE1-Mg-Lys in the β-CD complex (Figure 8).

[0221] 2D ROESY spectrum analysis was also performed to provide detailed information of the plausible host-guest interactions of RvE1-MgLys with β-CD (Figure 12), including the likely regions of intermolecular interactions shown by dashed rectangles and the specific interactions highlighted by solid rectangles. 2D ROESY analysis shows that some of the signals of RvE1-MgLys in the core region have spatial proximity of the interior protons of β-CD (3' and 5'), indicating the formation of a host-guest complex. Further analysis of the close-up of the intermolecular interactions of the 2D ROESY spectrum (Figure 13) was completed to determine the exact site and mode of inclusion, which showed a clear spatial proximity between the middle of RvE1 (protons 8-15) and the inside of the β-CD cavity (protons 3' and 5'). Based on these data, the plausible orientation of RvE1 inside the β-CD cavity is shown in Figure 23b.

[0222] Example 5: NMR characterization and molecular interaction studies of RvE1-MgLys complexed with γ-cyclodextrin

[0178] Figure 14 shows that the NMR signals of the γ-CD complex are seen in a well-defined region of the spectrum similar to the complexes of α-CD and β-CD.

[0223] As shown in Figure 7, the originally overlapping signals of 8 / 16 and 6 / 14 were partially and completely separated, respectively, indicating the complex formation between the unsaturated region of RvE1-MgLys and γ-CD. Similar to the complexes of α-CD and β-CD, the smallest signal shift was observed at the saturated portion of RvE1-Mg-Lys in the γ-CD complex (Figure 8).

[0224] 2D ROESY spectrum analysis was performed on the γ-CD complex to provide detailed information of the relevant host-guest interactions (FIG. 15). This analysis detected cross peaks between the signals of RvE1-MgLys and the core region of γ-CD in the designated region of intermolecular correlation shown by the dashed rectangle. The largest cross peak, which shows the correlation between the internal protons (3' and 5') of γ-CD and the signals of the unsaturated region of RvE1-MgLys, is highlighted by the solid rectangle. Further analysis of the close-up of the intermolecular interactions of the 2D ROESY spectrum (FIG. 16) was completed to determine the exact site and mode of encapsulation. The largest cross peaks in γ-CD can be observed between the 9, 10, and 15 protons of RvE1-MgLys (located at the center of the molecule) and the 3', 5', and 6' protons of γ-CD. An additional signal is between the same proton of γ-CD and the 12 and 20 protons of RvE1-MgLys. Based on these data, a reasonable orientation of RvE1 inside the γ-CD cavity is shown in Figure 23c.

[0225] Example 6: NMR characterization and molecular interaction studies of RvE1-MgLys complexed with 2-hydroxypropyl-β-cyclodextrin Based on FIG. 17, the NMR signals of the HP-β-CD complex can be grouped into three distinct regions. The anomeric protons appear between δ1H=5.0-5.2 ppm. Since HP-β-CD is a randomly substituted derivative of native CD, there are two anomeric signals in HP-β-CD, one representing the unsubstituted form and the other (usually with a higher chemical shift) representing the substituted form. The core of HP-β-CD is found between δ1H=3.5-4.2 ppm and contains the side chain (α' and β') signals. The side chain methyl (γ') protons establish a third group of signals for HP-β-CD above δ=1.0-1.2 ppm.

[0226] As shown in Figure 7, the slight signal change of RvE1-MgLys in the HP-β-CD complex compared to its spectrum as RvE1-MgLys alone indicates a chemical shift induced by the presence of HP-β-CD. The pattern is similar to that observed in the case of the β-CD system, but the most significant difference is the partial separation of the 7 / 15 signals and the partial overlap of the 11 / 17 signals.

[0227] As shown in the 2D ROESY spectrum analysis of the HP-β-CD complex in Figure 18, in the designated region of intermolecular correlations shown by the dashed rectangle, some cross peaks can be detected between the signal of RvE1-MgLys and the core region of HP-β-CD. Further analysis of the zoomed-in view of the intermolecular interactions in the 2D ROESY spectrum shows that the largest cross peaks are those of protons 9, 10, 12 and 18, with additional interactions detected at protons 6, 11 and 13 (Figure 19). Based on these data, it is clear that the observed intermolecular interactions are located inside HP-β-CD at the 3', 5' and 6' protons with a reasonable orientation shown in Figure 23d, indicating complex formation inside the cavity.

[0228] Example 7: NMR characterization and molecular interaction studies of RvE1-MgLys complexed with sulfobutyl ether β-cyclodextrin Based on FIG. 20, the SBE-β-CD complex 1 The H-NMR signals can also be grouped into three distinct regions. The anomeric protons appear between δ1H=5.0-5.2 ppm. As with HP-β-CD, SBE-β-CD has two anomeric signals, one representing the unsubstituted form and the other (usually with a higher chemical shift) representing the substituted form. The SBE-β-CD core is found between δ1H=3.5-4.1 ppm. The α' protons of the side chains overlap with the core region, and the remaining signals of the side chains contain two distinct regions, one between δ1H=2.9-3.0 ppm representing the δ' protons and the other between δ1H=1.7-1.9 ppm containing the middle protons, β' and γ' of the side chains.

[0229]

[0185] Intermolecular interactions are shown between the signal of RvE1-MgLys and the core region of SBE-β-CD, as shown by the dashed and solid rectangles in Figure 21. Further analysis of the close-up of the intermolecular interactions in the 2D ROESY spectrum (Figure 22) shows cross peaks in the region of the 3', 5', 6' and α' protons inside the SBE-β-CD cavity, with the appropriate orientation shown in Figure 23e.

[0230] Conjugate stability, pharmacokinetics and formulation studies Example 8: Stabilizing effect of cyclodextrins probed on RvE1-MgLys The stability of RvE1-MgLys alone ("RvE1 control") and RvE1-MgLys complexed with cyclodextrin ("RvE1 CD complex") was evaluated. Samples of the test material were placed in open tubes and kept exposed to air at 40°C and 75% relative humidity for 16 weeks. Quantitative demonstration of stability was determined using HPLC analytical method. Briefly, HPLC analysis was performed on a Halo Guard column (Halo C18 4.6 x 150 mm, 2.7 μm) mounted on an Agilent HPLC system equipped with a UV detector (272 nm detection). The mobile phase consisted of a gradient between solution A, water, and solution B, methanol, both of which contain 100 mM ammonium formate at pH 4.0. The gradient program was 95-0% for solution A and 5-100% for solution B. The flow rate was 0.9 mL / min over 50 min. Loss of stability was indicated by a decrease in area % purity (area %) of a single major peak of RvE1 (retention time 31.3 min) and the appearance of new HPLC peaks or an increase in area % of pre-existing impurities. All HPLC measurements of RvE1 purity and degradant levels were calculated based on area percent for detectable components of the molecule, including isomers and epimers of RvE1, impurities, and degradants induced by the test conditions.

[0231] At the first time point (baseline), RvE1 eluted as a single major peak with area % purity of 94% in the RvE1 control sample and 95.3%-95.5% in the RvE1 CD complex samples (Table 3). After 16 weeks of exposure to the above-mentioned test conditions, the RvE1 portion of the control group resulted in a significant decrease as indicated by the major peak decreasing to 80.7%. In contrast, the RvE1 portion of the various RvE1 CD complexes degraded to a much smaller extent under the same conditions.

[0232]

[0188] As shown in Table 3, the stabilizing effect of complexation with cyclodextrin can be expressed as the change in RvE1 purity, otherwise referred to as degradation rate, from baseline to week 16. The degradation rate of the RvE1 control at week 16 was 14.1% compared to an average degradation rate of 3.9% for the RvE1 CD complex. Thus, the degradation rate of RvE1 in the RvE1 CD complex was substantially reduced for each cyclodextrin tested.

[0233] [Table 3]

[0234] Importantly, the superior stability of the RvE1 CD complex was also associated with the formation of much smaller degradation peaks, greater than 0.2% or 1.0% at 8 and 16 weeks relative to the RvE1 control under the same conditions (Table 4). Notably, the RvE1 Mg-Lys γ-cyclodextrin complex showed the least number of degradation peaks greater than 0.2%. Overall, the improved stability of the RvE1 CD complex reduces the risk of loss of potency and / or emergence of unknown impurities, along with potential safety issues during manufacturing, formulation, packaging, storage, and distribution.

[0235] [Table 4]

[0236] Example 9: Long-term stabilizing effect of cyclodextrin on RvE1-MgLys and sodium RvE1

[0190] Chemical stability studies of RvE1-Mg-Lys alone and sodium RvE1 ("Na-RvE1") alone, as well as RvE1-Mg-Ly and sodium RvE1 complexed with cyclodextrin, were evaluated over a 24 week period. Samples of the test materials as solids were placed in open test tubes and maintained exposed to air at 40°C and 75% relative humidity. Quantitative demonstration of stability was determined using HPLC analytical methods as described above.

[0237] As shown in Table 5, at the first time point (baseline), RvE1 eluted as a single major peak with area % purity of 94.8% in the sample of RvE1-MgLys alone ("RvE1 MgLys control"), 96.2% in the sample of sodium RvE1 alone ("Na-RvE1 control"), and 95.9%-96.8% in the samples of cyclodextrin complexes formed from each RvE1 salt ("RvE1 CD complexes"). After 24 weeks of exposure to the test conditions described above, the RvE1 portion of the RvE1-MgLys control group resulted in a significant decrease as indicated by the major peak decreasing to 79.1% (consistent with the results shown in Table 3), and the RvE1 portion of the Na-RvE1 control group was completely degraded to 0%. In contrast, the RvE1 portion of the RvE1 CD complex was degraded to a much smaller extent under the same conditions. Importantly, the results shown in Table 5 indicate that the superior stability afforded by complexation with cyclodextrin can be achieved with different initial forms of the RvE1 moiety, in this example including either the sodium or magnesium lysinate salt of RvE1.

[0238]

[0192] At 24 weeks, the degradation rates of the RvE1-MgLys control and the Na-RvE1 control were 16.6% and 100%, respectively, compared to 0% for the RvE1-MgLys γ-cyclodextrin complex, 3.4% for the RvE1-MgLys HP-β-CD complex, and 4.3% for the Na-RvE1 HP-β-CD complex, consistent with the findings shown in Table 3.

[0239] [Table 5]

[0240] Example 10: Stabilizing effect of cyclodextrin on RvE1 salt in solution The chemical stability of RvE1-MgLys or Na-RvE1 alone and complexed with cyclodextrin in solution was evaluated. Samples of test material dissolved in deionized water at a concentration of 2 mg / ml RvE1 were placed in open test tubes and kept exposed to air at 40° C. and 75% relative humidity for 16 weeks. Quantitative demonstration of stability was determined using HPLC analysis as described above.

[0241] As shown in Table 6, at the first time point (baseline), RvE1 eluted as a single major peak in all samples with area % purity ranging from 94.8% to 96.2% in the samples of RvE1-MgLys alone ("RvE1-MgLys control") and Na-RvE1 alone ("Na-RvE1 control"), and 95.9% to 96.8% in the samples of RvE1 cyclodextrin complex ("RvE1 CD complex"). After 16 weeks of exposure to the test conditions described above, the RvE1 portion of the two control groups resulted in a significant decrease as indicated by the major peak, which decreased to 51.5% in the RvE1-MgLys control group and 18.6% in the Na-RvE1 control group. In contrast, the RvE1 portion of the RvE1 CD complex was degraded to a much smaller extent under the same conditions.

[0242]

[0195] At the same time, the excellent stability of the RvE1 CD complex in solution was unexpected, since there are examples of cyclodextrin compounds that have excellent stability as solids but poor stability in solutions, or better stability as solutions but poor stability as solids (Hamada Y et al., Chem Pharm Bull (Tokyo), 1975; 23(6): 1205-1211; Uekama, 2006; Popielec, 2017). Furthermore, the results of both RvE1 Mg-Lys and Na-RvE1 complexed with cyclodextrins show that enhanced stability can be achieved by different forms of the RvE1 moiety, in this case two RvE1 salts with very different stability profiles under accelerated conditions when not complexed with cyclodextrin.

[0243] [Table 6]

[0244] Example 11: Solid-state physical stability of RvE1 MgLysγ-cyclodextrin complex

[0196] The solid-state physical stability of RvE1 MgLys alone ("RvE1 control") and the RvE1-MgLys γ-cyclodextrin complex ("RvE1 complex") was evaluated (Figures 24A-D). Samples of the test materials as solids were placed in open test tubes under ambient conditions of 18-22°C temperature and 52%-72% relative humidity for 168 hours. Solid-state physical stability was visually assessed based on whether the material retained its form as a solid powder or transitioned to a liquid form.

[0245] At the first time point (baseline) (FIG. 24A), the RvE1 control and RvE1 complex samples were in powder form. At 8 hours (FIG. 24B), the RvE1-control and RvE1 complex samples maintained their powder form, while the RvE1 control sample began to aggregate and turn darker in color. At 24 and 168 hours (FIGS. 24C-D), the RvE1 control sample became a gel-like material, while the RvE1 complex sample maintained its form as a solid, off-white powder.

[0246] Example 12: Oral pharmacokinetics of RvE1 control versus RvE1 CD complex To establish the comparative oral pharmacokinetics ("PK") of RvE1 MgLys alone ("RvE1 control") and RvE1 MgLys complexed with γ-cyclodextrin ("RvE1 complex"), studies were conducted to investigate RvE1 exposure in plasma following a single dose of 4 mg / kg RvE1, RvE1 control or RvE1 complex administered via oral gavage at various time points for up to 8 hours. Quantitative measurements of RvE1 in plasma were determined by HPLC-MS / MS performed on a Halo Guard column (Halo C18 4.6×150 mm, 2.7 μm) attached to an Agilent HPLC system equipped with a UV detector (272 nm detection).

[0247] Sixty-five male C57B16 mice were randomized into three experimental groups at 8 weeks of age: baseline control ("baseline", n=5), RvE1 control (n=30), and RvE1 conjugate (n=30). Prior to administration of test substances, mice in the baseline group were sacrificed and blood was collected to establish endogenous levels of RvE1 in plasma. The remaining 60 mice received a single dose of RvE1 control or RvE1 conjugate. Five mice from each group were sacrificed at each of the following time points: 15 min, 30 min, 1 h, 2 h, 4 h, and 8 h after administration, allowing the determination of RvE1 concentration versus time curves. At the time of sacrifice, blood was collected into EDTA-treated tubes and immediately frozen.

[0248]

[0200] As shown in the PK curves and PK matrix summary in Table 7, the plasma PK of the RvE1 control group and the RvE1 conjugate group were comparable in terms of area under the cover (AUC, ng / ml*time), Cmax (ng / ml), and Tmax (min). Together, these data indicate that oral administration of the RvE1 conjugate can deliver comparable levels of RvE1 to the systemic circulation compared to the RvE1 control, thus supporting the use of cyclodextrin for pharmaceutical development of RvE1.

[0249] [Table 7]

[0250] Example 13: Formulation feasibility study of RvE1 MgLys γ-cyclodextrin complex in direct blends with pharmaceutical excipients

[0201] To determine the feasibility of preparing oral tablets using salts of RvE1 complexed with cyclodextrin, studies were conducted to examine the flow and compression properties of direct blends of RvE1 MgLys γ-cyclodextrin complex ("RvE1 complex") and standard pharmaceutical excipients shown in Table 8 ("Direct Blends") without granulation to improve flow.

[0251] [Table 8]

[0252]

[0202] In the flow portion of the testing, the direct blends without further granulation were first evaluated to determine particle size distribution ("PSD") using screen meshes ranging from #40 (coarse) to #325 (fine), corresponding to mesh diameters ranging from 400 to 44μ. As shown in Table 9, approximately 75% of the particles in the direct blends were 100 or greater in diameter (coarser) and approximately 25% were less than 100μ (finer). Once the particle size distribution of the direct blends was established, flow experiments were performed using a 16mm diameter orifice. In this portion of the testing, the direct blends without further granulation demonstrated adequate flow through a 16mm orifice, sufficient to allow gravity feeding through compression in a rotary press or direct filling into capsules.

[0253] [Table 9]

[0254]

[0203] In the compression portion of the study, the direct blend was compressed in a rotary press with an oval forming die (0.5457 cm x 2852 cm) at a compression force of approximately 100 bar to produce tablets with a thickness of approximately 5.8 mm (see Figures 25A-B). Tablets with a target total weight of 500 mg per tablet were prepared in two groups, low and high hardness (18 kilopounds (8.1 kg) and 30 kilopounds (13.6 kg), respectively). As shown in Table 10, both groups of tablets had minimal weight deviation and exhibited hardness, friability and disintegration time characteristics consistent with the target specifications.

[0255] [Table 10]

[0256] Equivalent

[0204] 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 following claims.

[0257]

[0205] All references described in this specification are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication or patent or patent application was specifically and individually intended to be incorporated by reference in its entirety for all purposes.

[0258]

[0206] The present invention should not be limited in scope by the specific embodiments described herein. Indeed, various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

Claims

1. A complex of Resolvin E1 (RvE1), or a salt, ester, or amide thereof, and γ-cyclodextrin (γ-CD), wherein the molar ratio of RvE1 to γ-CD in the complex is 1:

1.

2. The complex of claim 1 , wherein RvE1 is in the form of a salt.

3. 3. The complex of claim 2, wherein RvE1 is in the form of a salt of sodium, potassium, calcium, zinc, magnesium, or magnesium dilysinate.

4. The complex of claim 3, wherein RvE1 is RvE1 sodium salt.

5. RvE1 is a compound of formula I: 【Chemistry 1】 [In the formula, M is magnesium (Mg ２＋ ), calcium (Ca ２＋ ), and zinc (Zn ２＋ a divalent metal selected from A １ and A ２ are RvE1, respectively; R １ and R ２ each independently represents a C containing at least one basic functional group; １ ~C １０ is alkyl; X １ and X ２ are each independently H or CO-Z, and Z is a peptide containing 1 to 5 amino acids.

4. The complex of claim 3, in the form of a salt described below.

6. M is magnesium (Mg ２＋ ) or calcium (Ca ２＋ 6. The complex of claim 5, wherein the complex is selected from the group consisting of:

7. R １ and R ２ are each independently -(CH ２ ) ３ -Y １ and -(CH ２ ) ４ -Y ２ and Y １ and Y ２ is each selected from a positively charged primary amine, a positively charged secondary amine, a positively charged tertiary amine, and a positively charged guanidine.

8. X １ and X ２ 6. The complex of claim 5, wherein each is H.

9. R １ and R ２ are each -(CH ２ ) ４ -Y ２ and Y ２ Ga-NH ３ ＋ The complex of claim 5, wherein

10. M is magnesium (Mg ２＋ ) and R １ and R ２ are each -(CH ２ ) ４ -Y ２ and Y ２ Ga-NH ３ ＋ and X １ and X ２ are H, and A １ and A ２ is RvE1 and the resolvin of the complex is referred to as RvE1 magnesium dilysinate or "RvE1 MgLys".

11. 2. The complex of claim 1, wherein RvE1 is a sodium, potassium, calcium, zinc, or magnesium salt of RvE1.

12. A complex of RvE1 MgLys and gamma-cyclodextrin (gamma-CD), wherein the molar ratio of RvE1 to gamma-CD in the complex is 1:

1.

13. A complex of a sodium salt of RvE1 and gamma-cyclodextrin (gamma-CD), wherein the molar ratio of RvE1 to gamma-CD in the complex is 1:

1.

14. A pharmaceutical composition comprising the conjugate of any one of claims 1 and 11 to 13, and one or more pharmaceutically acceptable carriers and / or excipients.

15. 15. The pharmaceutical composition of claim 14 for treating an inflammatory disease or disorder of the gastrointestinal tract in a subject.

16. 16. The pharmaceutical composition of claim 15, wherein the inflammatory disease or disorder of the gastrointestinal tract is inflammatory bowel disease, ulcerative colitis, Crohn's disease, proctitis, pouchitis, eosinophilic colitis, lymphocytic colitis, collagenous colitis, diversion colitis, chemical colitis, ischemic colitis, infectious colitis, pseudomembranous colitis, indeterminate colitis, primary sclerosing cholangitis, gastritis, pancreatitis, viral gastroenteritis, NSAID enteropathy, irritable bowel syndrome, or postoperative bowel inflammation.

17. 15. The pharmaceutical composition of claim 14 for use in a method for treating cancer in a subject.

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