Carotenoid compositions and uses thereof

Abstract

The name of the invention is carotenoid compositions and uses thereof. Provided herein are pharmaceutical compositions comprising a carotenoid comprising a liposome encapsulating a carotenoid, including an ionizable carotenoid, such as trans-crocetin. The provided compositions have uses for the treatment of diseases, disorders and conditions associated with, but not limited to, infection, endotoxemia, inflammation, sepsis, ischemia, hypoxia, shock, stroke, lung injury, wound healing, traumatic injury, reperfusion injury, cardiovascular diseases, kidney diseases, liver diseases, inflammatory diseases, metabolic diseases, pulmonary disorders, and the like. Blood-related conditions and hyperproliferative diseases such as cancer. Methods of making, delivering, and using the pharmaceutical compositions are also provided.

Description

[0001] This application is a divisional application of the Chinese national phase patent application with application number 201980038129.2 after the PCT application with international application number PCT / US2019 / 030625, international application date May 3, 2019, and invention name “Carotenoid compositions and uses thereof” entered the Chinese national phase on December 7, 2020.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Application No. 62 / 666,699, filed May 3, 2018, and U.S. Provisional Application No. 62 / 809,123, filed February 22, 2019, each of which is incorporated herein in its entirety. Background Art

[0004] Under normal circumstances, bacteria and associated toxins (endotoxinemia) should not normally be found in the blood of healthy people. However, it is increasingly recognized that bacteria do enter the bloodstream under certain circumstances from sources such as the intestines (leaky gut syndrome) or the gingival interface (primarily in patients with poor gingival health). The consequence of this process, known as bacterial translocation, is a state of chronic bacteremia (bacteria) in the bloodstream. Bacterial translocation can lead to chronic bacteremia, which excretes endotoxins, such as bacterial lipopolysaccharides (LPS), causing a chronic low-grade chronic endotoxemia and the resulting chronic inflammatory state. This chronic low-grade inflammation associated with bacterial translocation has been implicated in the pathogenesis of many diseases.

[0005] More severe forms of endotoxemia are associated with sepsis, a life-threatening medical condition caused by a dysregulated host inflammatory response to infection. Sepsis is a global healthcare problem that affects an estimated 30 million people worldwide each year. The mortality rate from sepsis is approximately 40% in adults. Sepsis occurs when the body's attempt to fight infection causes the immune system to destroy tissues and organs. This uncontrolled response, which is normally intended to protect the body, causes widespread inflammation, vascular leakage, and abnormal blood clotting, leading to organ damage. In severe cases, blood pressure drops, multiple organ failure ensues, and the patient can quickly die from septic shock.

[0006] Management of sepsis is a complex and unmet clinical challenge that requires early recognition and management of infection, hemodynamic issues, and other organ dysfunction. The underlying infection in sepsis is treated with antimicrobial agents, most commonly broad-spectrum antibacterials, antivirals, and antifungals. Current treatment guidelines for the management of hemodynamic issues associated with sepsis and septic shock recommend the use of vasopressors and norepinephrine as first-line therapy.

[0007] Despite the measures discussed above, sepsis remains a major killer and new treatments for sepsis and its related medical conditions are still greatly needed. One observation is that most treatments for sepsis tend to address the infection and treatment of individual failing organ systems rather than the key underlying pathophysiological drivers of sepsis. An alternative approach would be to address the underlying mechanisms of sepsis in addition to treating the concomitant infection.

[0008] Sepsis, as well as many other medical conditions, are associated with oxygen deprivation (hypoxia). Globally, the leading cause of death is related to hypoxia in some way. Examples include, but are not limited to, coronary artery disease, stroke, chronic and acute respiratory diseases. In addition, hypoxia is a common feature of many cancers and leads to resistance to radiation, chemotherapy, and potentially immunotherapy. Preclinically, reversal of hypoxia in cancer is associated with improved response to treatment. This suggests that strategies to reverse hypoxia in the clinic may lead to improved cancer outcomes.

[0009] Carotenoids are a class of natural fat-soluble pigments found primarily in plants, where they act as accessory pigments and protect tissues through their ability to quench singlet oxygen and free radical species. Carotenoids are known to have antioxidant properties and thus provide many beneficial health effects, including reducing the potential risk of cardiovascular disease, cancer, and slowing and / or reversing the degenerative effects of aging on various human physiological activities. However, carotenoids are generally very lipophilic compounds, and the clinical use of many carotenoids is limited by their instability and low bioavailability.

[0010] Crocetin is a carotenoid with antioxidant properties, which is slightly soluble in water. Chemically speaking, crocetin is a 20-carbon apocarotenoid molecule, which contains seven double bonds and a carboxylic acid group at each end. It is reported that trans-crocetin (free acid) and its salt trans-sodium crocetin are used in free form (e.g., unencapsulated) pharmaceutical preparations, and are expected to treat the symptoms caused by hypoxia, ischemia and other medical conditions. However, they all do not show clinical therapeutic efficacy. This is partly due to the fact that the preparations of trans-crocetin and its sodium salt (trans-sodium crocetin) are so far limited by their instability, low bioavailability and short half-life.

[0011] In view of the health benefits conferred by carotenoids and the low bioavailability and instability outlined above, it is desirable to provide pharmaceutical compositions comprising carotenoids with improved bioavailability and stability. The compositions and methods provided address the shortcomings of the above-mentioned carotenoids. These compositions and methods will further help overcome the limitations of current treatment approaches for disease states associated with endotoxemia and hypoxia and other unmet medical needs. The compositions have applications as single agents and in combination with other therapies. Summary of the invention

[0012] The present disclosure provides a pharmaceutical composition comprising carotenoids, comprising liposomes encapsulating carotenoids, such as trans-crocetin, trans-norbixin and salts thereof. The provided composition has uses for treating diseases and disorders and conditions associated with, but not limited to, infection, inflammation, sepsis, ischemia, hypoxia, shock, stroke, injury, cardiovascular disease, kidney disease, liver disease, inflammatory disease, metabolic disease, lung disease, neurodegenerative disease, immune system disease, and hyperproliferative diseases such as cancer. Also provided are methods for preparing, delivering and using the pharmaceutical composition, and kits containing the composition.

[0013] The disclosed pharmaceutical composition provides enhanced delivery of carotenoids including ionizable polyene carotenoids (such as trans-crocetin) with poor pharmacokinetics and biodistribution. The present disclosure also provides liposome compositions showing high encapsulation efficiency (>98%), high drug-to-lipid ratio and / or enhanced drug retention. The provided pharmaceutical composition has uses for treating diseases and disorders and conditions associated with, but not limited to, infection, inflammation, sepsis, ischemia, hypoxia, anemia, trauma, injury, stroke, shock, diabetes, wound healing, injury (e.g., reperfusion injury, nerve injury, kidney injury, liver injury and lung injury) and hyperproliferative diseases (such as cancer), and conditions associated with the treatment of these diseases and conditions (e.g., anemia, neutropenia and immunosuppression). Methods for preparing, delivering and using the composition are also provided.

[0014] In some embodiments, the present disclosure provides:

[0015] [1] A pharmaceutical composition comprising an ionizable carotenoid salt having the formula: polyene carotenoid-Q, wherein:

[0016] The polyene carotenoids include

[0017] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds,

[0018] (b) a methyl or lower alkyl (C2-C3) substituent, and

[0019] (c) 1, 2, 3 or more ionizable groups; and

[0020] Q is a multivalent counter ion;

[0021] [2] A pharmaceutical composition comprising an ionizable carotenoid salt having the following formula:

[0022] Q-R1-polyene carotenoid-R2-Q, wherein

[0023] The polyene carotenoids include

[0024] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds,

[0025] (b) a methyl or lower alkyl (C2-C3) substituent, and

[0026] (c) 1, 2, 3 or more ionizable groups;

[0027] R1 and R2 are ionizable groups, such as the same ionizable group or different ionizable groups; and

[0028] Q is a multivalent counter ion;

[0029] [3] The pharmaceutical composition of [1] or [2], wherein the polyene carotenoid contains 1, 2, 3 or more anionic ionizable groups;

[0030] [4] The pharmaceutical composition according to [3], wherein the polyene carotenoid comprises at least one anionic ionizable group selected from the group consisting of a carboxyl group, a sulfonate group, a sulfate group, a phosphonate group, a phosphate group and a hydroxamate group;

[0031] [5] The pharmaceutical composition of [1] or [2], wherein the polyene carotenoid comprises 1, 2, 3 or more cationic ionizable groups (e.g., primary, secondary or tertiary amine groups, quaternary ammonium groups, choline groups, guanidine groups or imidazole groups);

[0032] [6] A pharmaceutical composition comprising an ionizable carotenoid salt having the formula: Q-trans-crocetin-Q, wherein

[0033] Q is a multivalent cationic counterion;

[0034] [7] A pharmaceutical composition comprising an ionizable carotenoid salt having the formula: Q-norbixin-Q, wherein:

[0035] Q is a multivalent cationic counterion;

[0036] [8] The pharmaceutical composition according to any one of [1] to [7], wherein the polyvalent counterion (Q) is a polyvalent cation (e.g., a divalent cation, such as a divalent metal cation or a divalent organic cation, or a trivalent cation, such as Fe 3+ );

[0037] [9] The pharmaceutical composition as described in [8], wherein the polyvalent cation is at least one selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2 + 、Co 2+ and Fe 2+ divalent cations; divalent organic cations, such as protonated diamines; or trivalent cations, such as Fe 3+ ;

[0038]

[10] The pharmaceutical composition according to [6], comprising magnesium trans-crocetin (MTC) or calcium trans-crocetin (CTC);

[0039]

[11] The pharmaceutical composition according to [7], comprising magnesium trans-norbixinate (MTN) or calcium trans-norbixinate (CTN);

[0040]

[12] A delivery vehicle comprising the pharmaceutical composition according to any one of [1] to

[11] ;

[0041]

[13] The delivery vehicle according to

[12] , which is a liposome;

[0042]

[14] A pharmaceutical composition comprising liposomes encapsulating an ionizable carotenoid salt having the following formula:

[0043] Polyene carotenoids-Q, wherein

[0044] The polyene carotenoids include

[0045] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds,

[0046] (b) a methyl or lower alkyl (C2-C3) substituent, and

[0047] (c) 1, 2, 3 or more ionizable groups; and

[0048] Q is (i) a multivalent counterion or (ii) a monovalent counterion;

[0049]

[15] A pharmaceutical composition comprising liposomes encapsulating an ionizable carotenoid salt having the formula:

[0050] Q-R1-polyene carotenoid-R2-Q, wherein

[0051] The polyene carotenoids include

[0052] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds,

[0053] (b) a methyl or lower alkyl (C2-C3) substituent, and

[0054] (c) 1, 2, 3 or more ionizable groups;

[0055] R1 and R2 are ionizable groups, such as the same ionizable group or different ionizable groups; and

[0056] Q is (i) a multivalent counterion or (ii) a monovalent counterion;

[0057]

[16] The pharmaceutical composition according to

[14] or

[15] , wherein the polyene carotenoid contains an anionic ionizable group;

[0058]

[17] The pharmaceutical composition of

[16] , wherein the polyene carotenoid comprises at least one anionic ionizable group selected from the group consisting of a carboxyl group, a sulfonate group, a sulfate group, a phosphonate group, a phosphate group and a hydroxamate group;

[0059]

[18] The pharmaceutical composition according to

[14] or

[15] , wherein the polyene carotenoid contains a cationic ionizable group (e.g., a primary, secondary or tertiary amine group, a quaternary ammonium group, a choline group, a guanidine group or an imidazole group),

[0060]

[19] A pharmaceutical composition comprising liposomes encapsulating an ionizable carotenoid salt having the formula:

[0061] Q-trans-crocetin-Q, wherein

[0062] Q is (i) a multivalent cationic counterion or (ii) a monovalent cation;

[0063]

[20] A pharmaceutical composition comprising liposomes encapsulating an ionizable carotenoid salt having the formula: Q-trans-norbixin-Q, wherein

[0064] Q is (i) a multivalent cationic counterion or (ii) a monovalent cation;

[0065]

[21] A pharmaceutical composition comprising liposomes encapsulating an ionizable carotenoid salt having the formula:

[0066] Q-R3-polyene carotenoid-R4-Q, wherein

[0067] The polyene carotenoid comprises 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds, optionally substituted with 1 to n methyl or lower C1-C3 alkyl substituents, wherein n=1 to 4; and

[0068] R3 and / or R4 are monocyclic and / or polar groups (e.g., the same or different monocyclic and / or polar groups); and

[0069] Q is (i) a multivalent counterion or (ii) a monovalent counterion;

[0070]

[22] A pharmaceutical composition comprising liposomes encapsulating a carotenoid having two linked polar groups, the polar groups may be the same or different polar groups, and the carotenoid has the following formula:

[0071] QA-polyene carotenoid-AQ, wherein

[0072] The polyene carotenoid comprises 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds, optionally substituted with 1 to n methyl or lower C1-C3 alkyl substituents, wherein n=1 to 4; and QA taken together and AQ taken together are monocyclic functional groups (e.g., the same or different monocyclic functional groups), the monocyclic functional groups are optionally selected from the monocyclic functional groups present in astaxanthin, lutein, lutein and zeaxanthin;

[0073]

[0074]

[23] The pharmaceutical composition according to any one of

[14] to

[21] , or wherein Q is a polyvalent counterion (e.g., a polyvalent cation, such as a divalent metal cation or a divalent organic cation);

[0075]

[24] The pharmaceutical composition as described in

[23] , wherein Q is at least one selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ divalent cations; divalent organic cations, such as protonated diamines; or trivalent cations, such as Fe 3+ ;

[0076]

[25] The pharmaceutical composition according to any one of

[14] to

[21] , wherein Q is a monovalent counterion (e.g., a monovalent metal cation or a monovalent organic cation);

[0077]

[26] The pharmaceutical composition as described in

[25] , wherein Q is at least one selected from NH4 + 、Na + , Li + and K + or an organic cation, such as a protonated amine;

[0078]

[27] The pharmaceutical composition according to

[19] , comprising magnesium trans-crocetin (MTC) or calcium trans-crocetin (CTC);

[0079]

[28] The pharmaceutical composition according to

[20] , comprising magnesium trans-norbixin (MTN) or calcium trans-norbixin (CTN);

[0080]

[29] The pharmaceutical composition according to any one of

[13] to

[28] , wherein the ionizable carotenoid / lipid ratio is 1 to 1000 g / mol, about 10 to 150 g / mol, about 20 to 100 g / mol, or any range therebetween;

[0081]

[30] The pharmaceutical composition according to any one of

[13] to

[29] , wherein the liposomes contain at least 0.1% to 97% by weight (w / w), or any range therebetween, of ionizable carotenoid;

[0082]

[31] The pharmaceutical composition according to any one of

[13] to

[30] , wherein the liposome has a diameter of 20 nm to 500 nm, 20 nm to 200 nm, or 80 nm to 120 nm, or any range therebetween;

[0083]

[32] The pharmaceutical composition according to any one of

[13] to

[31] , wherein the liposome is formed from a liposome component;

[0084]

[33] The pharmaceutical composition according to

[32] , wherein the liposome component comprises at least one of anionic lipids, cationic lipids and neutral lipids;

[0085]

[34] The pharmaceutical composition according to

[32] or

[33] , wherein the liposome component comprises at least one selected from the group consisting of: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol-PEG, and cholesterol-maleimide;

[0086]

[35] The pharmaceutical composition according to any one of

[13] to

[34] , wherein the liposomes contain oxidized phospholipids, such as OxPAPC;

[0087]

[36] The pharmaceutical composition according to

[35] , wherein the OxPAPC is an oxidized phospholipid containing a fragmented oxygenated sn-2 residue, an oxidized phospholipid containing a full-length oxygenated sn-2 residue, and / or an oxidized phospholipid containing a five-carbon sn-2 residue with an ω-aldehyde or ω-carboxyl group;

[0088]

[37] The pharmaceutical composition as described in

[35] , wherein the liposome comprises OxPAPC selected from the group consisting of HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC, 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxy-cyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxy-isoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine (PECPC), n-glyceryl-3-phosphocholine (PGPC); 1-palmitoyl-2-(9'oxonanoyl)-sn-glyceryl-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glyceryl-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glyceryl-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glyceryl-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glyceryl-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glyceryl-3-phosphocholine; or, the OxPAPC is a phospholipid containing epoxyisoprostaglandin;

[0089]

[38] The pharmaceutical composition according to

[37] , wherein the liposome contains PGPC;

[0090]

[39] The pharmaceutical composition of any one of

[13] -

[38] , wherein the liposomes comprise 0% to 100%, 0.1% to 30%, 1% to 25%, 5% to 20% or 7% to 15% OxPAPC (e.g., about 10% OxPAPC) or any range of OxPAPC therebetween;

[0091]

[40] The pharmaceutical composition according to any one of

[13] to

[39] , wherein the liposome comprises HSPE, cholesterol, PEG-DSPE-2000 and OxPAPC in a molar ratio of 2-5:1-4:0.01-0.3:0.05-1.5;

[0092]

[41] The pharmaceutical composition according to any one of

[13] to

[40] , wherein the liposome is pegylated;

[0093]

[42] The pharmaceutical composition according to any one of

[13] to

[41] , wherein one or more liposome components further comprises a steric stabilizer;

[0094]

[43] The pharmaceutical composition according to

[42] , wherein the steric stabilizer is at least one selected from the group consisting of: polyethylene glycol (PEG); poly-L-lysine (PLL); monosialoganglioside (GM1); poly(vinyl pyrrolidone) (PVP); poly(acrylamide) (PAA); poly(2-methyl-2-oxazoline); poly(2-ethyl-2-oxazoline); phosphatidyl polyglycerol; poly[N-(2-hydroxypropyl)methacrylamide]; amphiphilic poly-N-vinyl pyrrolidone; L-amino acid based polymers; oligoglycerols, copolymers containing polyethylene glycol and polypropylene oxide, poloxamer 188 and polyvinyl alcohol;

[0095]

[44] The pharmaceutical composition of

[43] , wherein the steric stabilizer is PEG, and the PEG has a number average molecular weight (Mn) of 200 to 5000 Daltons;

[0096]

[45] The pharmaceutical composition according to any one of

[13] to

[44] , wherein the liposome is anionic or neutral;

[0097]

[46] The pharmaceutical composition according to any one of

[13] to

[44] , wherein the liposome has a zeta potential of -150 to 150 mV, or -50 to 50 mV, or any range therebetween;

[0098]

[47] The pharmaceutical composition according to any one of

[13] to

[45] , wherein the liposome has a zeta potential less than or equal to zero (e.g., -150 to 0, or -50 to 0 mV, or any range therebetween);

[0099]

[48] ​​The pharmaceutical composition according to any one of

[13] to

[47] , wherein the liposome has a zeta potential greater than 0 (e.g., 0.2 to 150 mV, or 1 to 50 mV, or any range therebetween);

[0100]

[49] The pharmaceutical composition according to any one of

[13] to

[45] , or

[48] , wherein the liposome is cationic;

[0101]

[50] The pharmaceutical composition according to any one of [1] to

[49] , further comprising a pharmaceutically acceptable carrier;

[0102]

[51] The pharmaceutical composition of any one of [1] to

[50] , comprising a tonicity agent such as dextrose, mannitol, glycerol, potassium chloride or sodium chloride, optionally at a concentration greater than 0.1%, or at a concentration of 0.3% to 2.5%, or any range therebetween;

[0103]

[52] The pharmaceutical composition of

[51] , comprising trehalose or dextrose;

[0104]

[53] The pharmaceutical composition of

[52] , comprising 1% to 50% trehalose;

[0105]

[54] The pharmaceutical composition of

[51] , comprising dextrose, optionally 1% to 50% dextrose;

[0106]

[55] The pharmaceutical composition according to any one of [1] to

[54] , comprising 5% dextrose in a HEPES buffer solution;

[0107]

[56] The pharmaceutical composition of any one of [1] to

[55] , comprising a buffer such as HEPES buffered saline (HBS) or the like at a concentration of 1 to 200 mM and a pH of 2 to 8 or any range therebetween;

[0108]

[57] The pharmaceutical composition according to any one of [1] to

[56] , wherein the pharmaceutically acceptable carrier comprises a polyvalent metal acetate, such as magnesium acetate or calcium acetate, at a total concentration of 0.1 mM to 2000 mM, or 50 mM to 500 mM, or any range therebetween;

[0109]

[58] The pharmaceutical composition according to any one of [1] to

[57] , which has a pH value of 5-8, or a pH value of 6-7, or a pH value in any range therebetween;

[0110]

[59] The pharmaceutical composition of any one of

[13] -

[58] , wherein the liposomes contain less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, less than 10,000, or less than 5,000 ionizable carotenoid molecules;

[0111]

[60] The pharmaceutical composition according to any one of

[13] -

[59] , wherein the liposomes contain 10 to 100,000, 100 to 10,000, or 500 to 5,000, or any range therebetween, of ionizable carotenoid molecules;

[0112]

[61] The pharmaceutical composition according to any one of

[13] to

[60] , wherein the liposome further comprises a targeting moiety, and wherein the targeting moiety has a specific affinity for a surface antigen on a target cell of interest;

[0113]

[62] The pharmaceutical composition of

[61] , wherein the targeting moiety is attached to one or both of PEG and the exterior of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the exterior of the liposome via a covalent bond;

[0114]

[63] The pharmaceutical composition of any one of

[61] or

[62] , wherein the targeting moiety is a polypeptide;

[0115]

[64] The pharmaceutical composition according to any one of

[61] to

[63] , wherein the targeting moiety is an antibody or an antigen-binding fragment of an antibody;

[0116]

[65] A pharmaceutical composition according to any one of

[61] to

[64] , wherein the targeting moiety binds to the surface antigen, such as using The equilibrium dissociation constant (Kd) was determined to be 50 x 10 -12 Up to 10x 10 -6 within the scope;

[0117]

[66] The pharmaceutical composition according to any one of

[61] to

[65] , wherein the targeting moiety specifically binds to one or more folate receptors selected from the group consisting of folate receptor α (FR-α), folate receptor β (FR-β) and folate receptor δ (FR-δ);

[0118]

[67] The pharmaceutical composition according to any one of

[61] to

[66] , wherein the targeting moiety comprises one or more selected from the group consisting of an antibody, a humanized antibody, an antigen-binding fragment of an antibody, a single-chain antibody, a single-domain antibody, a bispecific antibody, a synthetic antibody, a pegylated antibody, and a multimeric antibody;

[0119]

[68] The pharmaceutical composition according to any one of

[62] to

[67] , wherein each PEGylated liposome comprises 1 to 1000, 50 to 750, 100 to 500, or 30 to 200 targeting moieties or any range therebetween;

[0120]

[69] The pharmaceutical composition according to any one of

[13] to

[67] , further comprising one or more of FABP, an immunostimulatory agent, an immunosuppressive agent, a detectable marker and a maleimide, wherein the FABP, the immunostimulatory agent, the immunosuppressive agent, the detectable marker or the maleimide is attached to the PEG or the outside of the liposome;

[0121]

[70] The pharmaceutical composition according to

[69] , comprising at least one immunostimulant selected from the group consisting of a protein immunostimulant, a nucleic acid immunostimulant, a chemical immunostimulant, a hapten, and an adjuvant;

[0122]

[71] The pharmaceutical composition of any one of

[69] or

[60] , wherein the immunostimulant is at least one selected from the group consisting of: fluorescein; fluorescein isothiocyanate (FITC); DNP; β-glucan; β-1,3-glucan; β-1,6-glucan; a resolvin (e.g., a resolvin D, such as Dn-6DPA or Dn-3DPA, a resolvin E, or a T-series resolvin); and a Toll-like receptor (TLR) modulator, such as oxidized low-density lipoprotein (e.g., OXPAC, PGPC) or an eritoran-like lipid (e.g., E5564);

[0123]

[72] The pharmaceutical composition according to any one of

[69] to

[71] , comprising FABP;

[0124]

[73] The pharmaceutical composition according to any one of

[69] to

[72] , further comprising a hapten;

[0125]

[74] The pharmaceutical composition of

[73] , wherein the hapten comprises one or more of fluorescein or β1,6-glucan;

[0126]

[75] The pharmaceutical composition according to any one of [1] to

[74] , further comprising at least one cryoprotectant selected from the group consisting of: mannitol; trehalose; sorbitol and sucrose;

[0127]

[76] A targeted composition comprising the pharmaceutical composition according to any one of [1] to

[75] ;

[0128]

[77] A non-targeted liposome composition comprising the pharmaceutical composition according to any one of

[13] -

[60] or

[69] -

[75] ;

[0129]

[78] The pharmaceutical composition according to any one of [1] to

[77] , which is used to treat a disease or condition in a subject;

[0130]

[79] Use of the pharmaceutical composition according to any one of [1] to

[78] in the manufacture of a medicament for treating a disease in a subject;

[0131]

[80] A method for treating or preventing a disease in a subject in need of such treatment or prevention, the method comprising administering to the subject the pharmaceutical composition of any one of [1] to

[79] ;

[0132]

[81] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is related to endotoxemia;

[0133]

[82] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is sepsis;

[0134]

[83] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the subject is a burn victim;

[0135]

[84] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is an infection (e.g., a bacterial infection such as Pseudomonas aeruginosa infection, Staphylococcus aureus infection (e.g., MRSA) or a condition associated therewith, or an enterococcal infection (e.g., VRE), a fungal infection (e.g., a Candida infection (e.g., invasive candidiasis) or a condition associated therewith, or a parasitic infection or a condition associated therewith, such as malaria (or a condition associated therewith, such as cerebral malaria, severe anemia, acidosis, acute renal failure and ARDS), schistosomiasis and human African trypanosomiasis, and a condition associated therewith; a viral infection or a condition associated therewith, such as Ebola virus, dengue fever and Marburg virus (or a condition associated therewith, such as influenza, measles and viral hemorrhagic fever));

[0136]

[85] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is bacteremia;

[0137]

[86] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is a liver disease or condition (e.g., cirrhosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH); alcoholic liver disease, acute liver injury and cirrhosis);

[0138]

[87] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is a pulmonary disease or condition (e.g., acute respiratory distress syndrome (ARDS), pulmonary fibrosis, pulmonary hemorrhage, lung injury, lung cancer, chronic obstructive pulmonary disease (COPD) and other respiratory disorders);

[0139]

[88] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is a kidney disease (e.g., lipopolysaccharide drug or toxin-induced acute kidney injury (AKI) and end-stage renal disease);

[0140]

[89] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is an autoimmune disorder (e.g., psoriasis, cystic fibrosis and rheumatoid arthritis);

[0141]

[90] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is sclerosis (e.g., systemic sclerosis);

[0142]

[91] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the subject is a critically ill patient;

[0143]

[92] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the subject is at risk of developing sepsis;

[0144]

[93] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is a low-grade endotoxemia disease;

[0145]

[94] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is inflammation (e.g., systemic inflammation, low-grade inflammation, acute inflammation and chronic inflammatory disease);

[0146]

[95] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is inflammatory bowel disease (e.g., Crohn's disease and ulcerative colitis);

[0147]

[96] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the subject is immunocompromised;

[0148]

[97] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the subject is receiving chemotherapy and / or is immunosuppressed (e.g., a patient with febrile neutropenia);

[0149]

[98] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is a metabolic disease;

[0150]

[99] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is insulin resistance;

[0151]

[100] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is diabetes or a related condition, such as gangrene, diabetic necrosis, diabetic neuropathy, diabetic vascular disease (e.g., microvascular disease, such as retinopathy and nephropathy, and diabetic ulcers);

[0152]

[101] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is type 2 diabetes;

[0153]

[102] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is a cardiovascular disease (e.g., coronary artery disease such as myocardial infarction, sudden cardiac death, cardiopulmonary arrest, hypertension, pulmonary hypertension, atherosclerosis, occlusive arterial disease, Raynaud's disease, peripheral vascular disease, other vascular diseases such as Buerger's disease, Takayasu's arthritis and post-cardiac arrest syndrome (PCAS), chronic venous insufficiency, heart disease, congestive heart failure, chronic skin ulcers);

[0154]

[103] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is characterized by ischemia or hypoxia (e.g., ischemia-reperfusion injury, transient cerebral ischemia, cerebral ischemia-reperfusion, ischemic stroke, hemorrhagic stroke, traumatic brain injury, migraine (e.g., chronic migraine or severe migraine), gastrointestinal ischemia, kidney disease, pulmonary embolism, acute respiratory failure, neonatal respiratory distress syndrome, obstetric emergencies for reducing perinatal complications (such as pre-eclampsia / eclampsia and conditions leading to cerebral palsy), myocardial infarction, acute limb or mesenteric ischemia, cardiac cirrhosis, chronic peripheral vascular disease, congestive heart failure, atherosclerotic stenosis, anemia, thrombosis, embolism, macular degeneration, neurodegenerative diseases (such as Alzheimer's disease, Parkinson's disease or amyotrophic lateral sclerosis (ALS)), sleep apnea, and surgical or traumatic injury);

[0155]

[104] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is a heart attack or a stroke (e.g., ischemic and hemorrhagic stroke);

[0156]

[105] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is shock (e.g., cardiogenic shock, hypovolemic shock, septic shock, neurogenic shock and anaphylactic shock);

[0157]

[106] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is associated with nitric oxide deficiency (e.g., sickle cell disease, paroxysmal nocturnal hemoglobinuria (PNH), hemolytic anemia, thalassemia, another red blood cell disorder, purpura (such as thrombotic thrombocytic purpura (TTP)), hemolytic uremic syndrome (HUS), idiopathic thrombocytopenia (ITP), another platelet disorder, coagulation disorder (such as disseminated intravascular coagulopathy (DIC)), purpura fulminans, heparin-induced thrombocytopenia (HIT), leukocytosis and hyperviscosity syndrome, or a condition related thereto);

[0158]

[107] The pharmaceutical composition of

[78] , the use of

[79] or the method of

[80] , wherein the disease or condition is endotoxemia, such as endotoxemia associated with a condition such as periodontal disease (e.g., periodontitis or gingival inflammation), chronic alcoholism, chronic smoking, transplantation, neonatal necrotizing enterocolitis or neonatal ear infection;

[0159]

[108] A method for reducing systemic levels of LPS, endotoxin and / or another systemic inflammatory trigger in a subject in need thereof, the method comprising administering to the subject the pharmaceutical composition of any one of [1]-

[78] ;

[0160]

[109] The method according to any one of

[80] -

[108] , wherein the pharmaceutical composition is administered as a combination therapy with another therapeutic agent;

[0161]

[110] A method for preparing a liposome composition as described in any one of

[13] to

[77] , the method comprising: forming a mixture comprising: a liposome component in a solution; homogenizing the mixture to form liposomes in the solution; and processing the mixture to form liposomes containing ionizable carotenoids;

[0162]

[111] The method according to

[110] , wherein the processing step comprises one or more of the following steps: thin film hydration, extrusion, online mixing, ethanol injection technology, freeze-thaw technology, reverse phase evaporation, dynamic high pressure microfluidization, microfluidic mixing, double emulsification, freeze-dried double emulsification, 3D printing, membrane contactor method and stirring;

[0163]

[112] The method of

[110] or

[111] , wherein the processing step comprises one or more steps of improving the size of the liposomes by one or more of extrusion, high pressure microfluidization and / or ultrasound steps;

[0164]

[113] A method for preparing a pharmaceutical composition, the method comprising:

[0165] (a) preparing a liposome solution comprising liposomes in a weak acid salt of a multivalent metal;

[0166] (b) adding the ionizable carotenoid according to any one of [1] to [7],

[10] or

[11] to the liposome solution; and

[0167] (c) maintaining the ionizable carotenoid in the liposome solution for a sufficient time to load the ionizable carotenoid into the liposomes;

[0168]

[114] The method of

[113] , wherein the weak acid is an organic acid (e.g., an organic acid selected from acetic acid, gluconic acid, tartaric acid, glutamic acid, citric acid, formic acid and glycine);

[0169]

[115] The method of

[113] or

[114] , wherein the selected polyvalent metal is a divalent metal (e.g., selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ divalent metals) or trivalent metals such as Fe 3+ ;

[0170]

[116] The method according to any one of

[113] to

[115] , wherein the weak acid is acetic acid and the polyvalent metal is Ca 2+ or Mg 2+ (i.e., the weak acid salt of the multivalent metal is calcium acetate or magnesium acetate, respectively);

[0171]

[117] A pharmaceutical composition prepared according to the method of any one of

[109] to

[115] ;

[0172]

[118] A method for preparing a pharmaceutical composition comprising liposomes encapsulating trans-crocetin, the method comprising:

[0173] (a) preparing a liposome solution containing liposomes and a solution containing a weak acid salt of a multivalent metal;

[0174] (b) adding trans-crocetin to the liposome solution; and

[0175] (c) maintaining the trans-crocetin in the liposome solution for a sufficient time to load the trans-crocetin into the liposomes;

[0176]

[119] The method of

[118] , wherein the weak acid is an organic acid (e.g., acetic acid, gluconic acid, tartaric acid, glutamic acid, citric acid, formic acid, and glycine);

[0177]

[120] The method of

[118] or

[119] , wherein the polyvalent metal is a divalent metal (e.g., selected from Ca 2 + Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ divalent metals) or trivalent metals such as Fe 3+ ; and / or

[0178]

[121] The method of any one of claims

[118] to

[120] , wherein the weak acid is acetic acid and the polyvalent metal is Ca 2+ or Mg 2+ ; and / or

[0179]

[122] A pharmaceutical composition prepared according to the method described in any one of

[117] to

[120] .

[0180] Still other features and advantages of the compositions and methods described herein will become more apparent from the following detailed description when read in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0181] Figure 1A , Figure 1B , Figure 1C as well as Figure 1D Exemplary ionizable polyene carotenoids of provided pharmaceutical compositions are shown.

[0182] Figure 2. Stability of trans-calcium crocetin liposomes (CTC-LP) over 6 months at 4°C. The CTC-LP test preparations contained drug / lipid (D / L) ratios of 80, 60, and 40. Each CTC-LP test preparation showed negligible leaching (change in D / L ratio) over the 6-month evaluation period.

[0183] Figure 3 Liposomal CTC batch reproducibility. Four batches of liposomal CTC were reproducible and stable at 4°C for at least 7 months.

[0184] Figure 4 Stability of trans-magnesium crocetin liposomes (MTC-LP) over 6 months at 4°C. The MTC-LP test preparations contained drug / lipid (D / L) ratios of 80, 60, and 40. Each MTC-LP test preparation showed negligible leaching (change in D / L ratio) over the 2 month evaluation period.

[0185] Figure 5 .Survival study 1 (TP-936): Study of the efficacy of CTC-LP in the mouse CLP sepsis model. Survival curves of mice treated with the following test articles: (a) liposomal CTC (D / L80) + antibiotics, (b) liposomal CTC (D / L80) and PGPC + antibiotics, (c) saline + antibiotics and (d) sham treatment (sham). The treatment groups of test articles (a) and (b) (in combination with imipenem) showed a trend of reduced mortality when compared with the control group (c) treated with imipenem.

[0186] Figure 6 Survival Study 2 (TP-967): Mouse CLP Sepsis Study. Survival curves of mice treated with the following test articles: (a) liposomal PGPC + antibiotics, (b) liposomal (PGPC and CTC) (D / L80) + antibiotics, (c) liposomal CTC (D / L80) + antibiotics, and (d) saline + antibiotics. The treatment group of the test article (c) showed a trend of reduced mortality when compared to the control group (d) treated with imipenem.

[0187] Figure 7Survival Study 3 (TP-986): Mouse CLP Sepsis Study. Survival curves of mice treated with the following test articles: (a) Liposomal CTC (D / L80) (1 mg / kg) + antibiotics, (b) Liposomal CTC (D / L80) (5 mg / kg) + antibiotics, (c) Liposomal CTC (D / L80) (25 mg / kg) + antibiotics, (d) Liposomal CTC (D / L80) (50 mg / kg) + antibiotics and (e) Saline + antibiotics. The test article control groups (a), (c) and (d) all showed a trend towards reduced mortality when compared to the imipenem-treated control group (d). The test article treatment group (b) (Liposomal CTC (D / L80) (5 mg / kg) + antibiotics) showed a statistically significant reduction in mortality when compared to the imipenem-treated control group (d) (P = 0.0321). DETAILED DESCRIPTION

[0188] Applicants have surprisingly discovered that pharmaceutical compositions such as liposomes comprising multivalent ionizable carotenoid salts containing multivalent counterions can significantly improve pharmacokinetics (e.g., half-life, stability, and bioavailability) and can dramatically increase drug exposure by providing a sustained release of the ionizable carotenoid when compared to, for example, carotenoid free acids and ionizable carotenoid salts containing monovalent counterions.

[0189] definition

[0190] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art to which the present disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the implementation or testing of the provided compositions, suitable methods and materials are described below. Each publication, patent application, patent and other references mentioned herein are incorporated herein by reference in their entirety. In the event of a conflict, the present specification including the definitions shall prevail. In addition, materials, methods and examples are illustrative only and are not intended to be limiting.

[0191] Other features and advantages of the disclosed compositions and methods will be apparent from the following disclosure, drawings, and claims.

[0192] It should be understood that regardless of the embodiments described herein using the language "comprising", otherwise similar embodiments described with the terms "containing", "consisting of", and / or "consisting of essentially" are also provided. However, when used as a transitional phrase in the claims, each should be interpreted individually and in the appropriate legal and factual context (e.g., in the claims, the transitional phrase "comprising" is considered more of an open phrase, while "consisting of" is more closed, and "consisting essentially of" is in an intermediate zone).

[0193] As used herein, the singular forms "a", "an", and "the" include the plural forms unless explicitly stated or it is clear from the context that this is not intended. The singular forms "a", "an", and "the" also include the statistical average composition, characteristics, or size of particles in a particle population (e.g., average liposome diameter, average liposome zeta potential, average number of targeting moieties on liposomes in a liposome solution, average number of encapsulated carotenoids). The average particle size and zeta potential of liposomes in a pharmaceutical composition can be routinely measured using methods known in the art such as dynamic light scattering. The average amount of therapeutic agent in a nanoparticle composition can be routinely measured, for example, using absorption spectroscopy (e.g., UV-visible spectroscopy).

[0194] As used herein, the terms "approximately" and "about", when applied to one or more values ​​of interest, refer to values ​​that are similar to the stated reference values. In certain embodiments, the terms "approximately" or "about" refer to a range of values ​​that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less of either direction (greater or less than the stated reference value) of the stated reference value, unless otherwise specified or otherwise apparent from the context (unless such numbers would exceed 100% of the possible values). For example, when used in the context of the amount of a given compound in the lipid component of a nanoparticle composition, "about" may mean ±10% of the recited value. For example, a nanoparticle composition comprising a lipid component having about 40% of a given compound may comprise 30-50% of the compound.

[0195] The term "and / or", as used herein in phrases such as "A and / or B", is intended to include both A and B; A or B; A (alone); and B (alone). Likewise, the term "and / or", as used in phrases such as "A, B, and / or C", is intended to include each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0196] Where embodiments of the present disclosure are described in terms of Markush groups or other groupings of alternatives, the disclosed compositions or methods include not only the entire group listed as a whole, but also each member of the group listed individually and all possible subgroups of the main group, and also include the main group lacking one or more group members. The disclosed compositions and methods also contemplate the explicit exclusion of one or more of any group members in the disclosed compositions or methods.

[0197] The term "liposome" refers to a closed vesicle with an internal phase (i.e., an internal space (internal solution)) enclosed by a lipid bilayer. Liposomes can be small single-film liposomes such as small unilamellar vesicles (SUV), large single-film liposomes such as large unilamellar vesicles (LUV), larger single-film liposomes such as giant unilamellar vesicles (GUV), multilamellar liposomes with multiple concentric membranes (e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10 concentric membranes) such as multilamellar vesicles (MLV), or liposomes with multiple irregular and non-concentric membranes such as multivesicles (MVV). Liposomes and liposome preparations are well known in the art. Lipids capable of forming liposomes include all substances with fat or fat-like properties. Lipids that can constitute the lipids in liposomes include, but are not limited to, glycerides, glycerophospholipids, glycerophosphinolipids, glycerophosphonolipids, sulfatides, sphingolipids, phospholipids, isoprenolides, steroids, stearin, sterols, archeolipids, synthetic cationic lipids, and carbohydrate-containing lipids.

[0198] A "liposomal composition" is a prepared composition comprising liposomes and contents within the liposomes, particularly lipids that form one or more liposomal bilayers, compounds other than lipids within one or more bilayers of the liposomes, compounds within and associated with one or more aqueous interiors of the liposomes, and compounds bound to or associated with the outer layer of the liposomes. Thus, in addition to the lipids of the liposomes, the liposomal compositions described herein may suitably include, but are not limited to, therapeutic agents, immunostimulants, vaccine antigens and adjuvants, excipients, carriers, and buffers. In preferred embodiments, such compounds complement or do not significantly impair the stability or AGP incorporation efficiency of the liposomal composition.

[0199] The terms liposome "inner phase", "inner space" and "inner core" are used interchangeably to refer to the aqueous region enclosed within (i.e., encapsulated by) the lipid bilayer of the liposome. The solution of the liposome inner phase is referred to as the "inner solution". In contrast, the term "liposome outer phase" refers to the region not enclosed by the lipid bilayer of the liposome, such as the region outside the inner phase and the lipid bilayer when the liposome is dispersed in a liquid.

[0200] The term "counterion" refers to an anionic or cationic counterion. A "cationic counterion" is a positively charged atom or group that associates with an anionic atom or group to maintain electrical neutrality. Exemplary cationic counterions include inorganic cations (e.g., metal cations (e.g., alkali metal cations, alkaline earth metal cations, and transition metal cations)) and organic cations (e.g., ammonium cations, sulfonium cations, phosphonium cations, and pyridinium cations). An "anionic counterion" is a negatively charged atom or group that associates with a cationic atom or group to maintain electrical neutrality. Exemplary anionic counterions include halide anions (e.g., F - 、Cl-、Br - and I - )、NO3 - 、ClO4 - OH - 、H2PO4 -2 、HSO4 - , sulfonate anions (e.g., methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphorsulfonate, naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate, ethane-1-sulfonic acid-2-sulfonate, etc.), and carboxylate anions (e.g., acetate, ethanoate, propionate, benzoate, glycerate, lactate, tartrate, and glycolate). The counterion may be monovalent or polyvalent (e.g., divalent, trivalent, tetravalent, etc.).

[0201] The term "ionizable" refers to compounds containing at least one functional group that (a) is positively or negatively charged (i.e., "ionizes") and thus associates with a counterion of the opposite charge, or (b) is neutral but ionizes at higher or lower pH values. Thus, ionizable compounds include quaternary ammonium salts as well as uncharged amine and carboxylate moieties and uncharged carboxyl groups.

[0202] As used herein, the term "carotenoid" refers to an organic pigment that is structurally composed of a polyolefin chain and that may terminate in a ring. Carotenoids are divided into two categories: xanthophylls (which contain oxygen atoms) and carotenes (which do not contain oxygen atoms). Figures 1A to 1D Non-limiting examples of carotenoids suitable for use in the provided compositions and methods are provided in. Carotenoids with ionizable functional groups include naturally occurring carotenoid sulfates, carotenoid carboxylates / carboxylates, synthetic phosphates, blue carotenoid oxonium ions, and blue carotenoid proteins.

[0203] As used herein, the term "polyene carotenoid" refers to a carotenoid containing 3 or more conjugated double bonds and methyl or lower alkyl (C2-C3) substituents.

[0204] The term "naturally occurring" refers to a compound or composition that occurs in nature, regardless of whether the compound or composition is isolated from a natural source or chemically synthesized. Examples of naturally occurring carotenoid mono- and dicarboxylic acids include crocetin, norbixin, erucicin, and neurosporaxanthin.

[0205] "Apocarotenoids" are carotenoid degradation products in which the normal structure has been shortened (e.g., C40) by removal of fragments from one or both ends. Examples of naturally occurring apocarotenoids include crocetin (C20), annatto (C25), vitamin A, abscisic acid, mycorradicin, and blumenin.

[0206] The term "targeting moiety" as used herein refers to a molecule that provides enhanced affinity to a selected target (e.g., a cell, cell type, tissue, organ, region of the body) or compartment (e.g., a cell, tissue or organ compartment). The targeting moiety may comprise a variety of entities. The targeting moiety may comprise a naturally occurring molecule, or a recombinant or synthetic molecule. In some embodiments, the targeting moiety is an antibody, an antigen-binding antibody fragment, a bispecific antibody, or other antibody-based molecules or compounds. In some embodiments, the targeting moiety is an aptamer, an avimer, a receptor binding ligand, a nucleic acid, a biotin-avidin binding pair, a peptide, a protein, a carbohydrate, a lipid, a vitamin, a toxin, a component of a microorganism, a hormone, a receptor ligand, or any derivative thereof. Other targeting moieties are known in the art and are encompassed by the present disclosure.

[0207] The term "specific affinity" or "specifically binds" means that a targeting moiety (such as an antibody or antigen-binding antibody fragment) reacts or associates with an epitope, protein or target molecule more frequently, more rapidly, with a longer duration, with a greater affinity, or with some combination of the above characteristics, compared to alternative substances including proteins unrelated to the target epitope. Due to the sequence identity between homologous proteins in different species, specific affinity can include binding agents that recognize proteins or targets in more than one species in several embodiments. Similarly, due to homology within certain regions of polypeptide sequences of different proteins, the term "specific affinity" or "specifically binds" can include binding agents that recognize more than one protein or target. It should be understood that in some embodiments, a binding moiety that specifically binds to a first target may or may not specifically bind to a second target. Therefore, "specific affinity" does not necessarily require (although it can include) exclusive binding, for example, binding to a single target. Therefore, a targeting moiety can specifically bind to more than one target in some embodiments. In some embodiments, multiple targets can be bound by the same targeting moiety.

[0208] The term "epitope" refers to that portion of an antigen that can be recognized and specifically bound by a targeting moiety (i.e., a binding moiety) such as an antibody. When the antigen is a polypeptide, the epitope can be formed by adjacent amino acids and non-adjacent amino acids juxtaposed by the tertiary folding of the protein. Epitopes formed by adjacent amino acids are usually retained after protein denaturation, while epitopes formed by tertiary folding are usually lost after protein denaturation. Epitopes usually include at least 3, and more commonly, at least 5 or 8-10 amino acids with a unique spatial conformation.

[0209] Expressions like "binding affinity for a target", "binding to a target", and similar expressions known in the art refer to properties of a targeting moiety that can be directly measured by determining an affinity constant, e.g., the amount of associated and dissociated targeting moiety at a given antigen concentration. Different methods can be used to characterize molecular interactions, such as, but not limited to, competition assays, equilibrium assays, and microcalorimetric assays, as well as real-time interaction assays based on surface plasmon resonance interactions (e.g., using Instruments). These methods are well known to those skilled in the art and are described, for example, in Neri et al., Tibtech 14:465-470 (1996) and Jansson et al., J. Biol. Chem. 272:8189-8197 (1997).

[0210] As used herein, "effective amount" refers to the dosage of an agent sufficient to provide a medically desired result. The effective amount will vary with the desired outcome, the specific condition being treated or prevented, the age and physical condition of the subject being treated, the severity of the condition, the duration of treatment, the nature of concurrent therapy or combined therapy (if any), the specific route of administration, and similar factors within the knowledge and expertise of health practitioners. "Effective amount" can be determined empirically and in a conventional manner relative to the stated purpose. In the case of cancer, the effective amount of the agent can reduce the number of cancer cells; reduce tumor size; inhibit (i.e., slow down and preferably stop to some extent) cancer cells from infiltrating into surrounding organs; inhibit (i.e., slow down and preferably stop to some extent) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with the disease to some extent. To some extent, the drug may prevent growth and / or kill existing cancer cells, it may have a cell growth inhibitory effect and / or cytotoxicity. For cancer therapy, in vivo efficacy can be measured, for example, by assessing the duration of survival, the duration of progression-free survival (PFS), the response rate (RR), the duration of response, and / or the quality of life.

[0211] The terms "hyperproliferative disorder," "proliferative disease," and "proliferative disorder" are used interchangeably herein and refer to unwanted or uncontrolled cell proliferation of excessive or abnormal cells that is undesirable, such as neoplastic or hyperplastic growth, occurring in vivo or in vitro. In some embodiments, the proliferative disease is a cancer or tumor disease (including benign or cancerous) and / or any metastasis, regardless of where the cancer, tumor, and / or metastasis is located. In some embodiments, the proliferative disease is a benign or malignant tumor. In some embodiments, the proliferative disease is a non-cancerous disease. In some embodiments, the proliferative disease is a hyperproliferative condition, such as hyperplasia, fibrosis (particularly pulmonary fibrosis, but also other types of fibrosis, such as renal fibrosis), angiogenesis, psoriasis, atherosclerosis, and vascular smooth muscle proliferation, such as stenosis or restenosis after angioplasty.

[0212] "Cancer," "tumor," or "malignancy" are used as synonymous terms and refer to any of a number of diseases characterized by uncontrolled, abnormal cell proliferation, the ability of affected cells to spread locally or through the bloodstream and lymphatic system to other parts of the body (metastasis), and any of a number of characteristic structural and / or molecular features. "Tumor," as used herein, refers to all neoplastic cell growth and proliferation, whether malignant or benign, and all precancerous and cancerous cells and tissues. "Cancerous tumor" or "malignant cell" is understood to be a cell with specific structural characteristics, lack of differentiation, and capable of infiltration and metastasis. Cancers that can be treated using the carotenoid pharmaceutical compositions provided herein include, but are not limited to, non-hematologic malignancies, including, for example, lung cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, head and neck cancer, gastric cancer, gastrointestinal cancer, colorectal cancer, esophageal cancer, cervical cancer, liver cancer, kidney cancer, bile duct cancer, gallbladder cancer, bladder cancer, sarcoma (e.g., osteosarcoma), brain cancer, central nervous system cancer, and melanoma; and hematologic malignancies, such as, for example, leukemia, lymphoma and other B-cell malignancies, myeloma and other plasma cell dysplasia or atypia. Other types of cancers and tumors that can be treated using trans-crocetin compositions are described herein or are otherwise known in the art. The terms "cancer," "cancerous," "cell proliferative disorders," "proliferative disorders," and "tumors" are not mutually exclusive herein.

[0213] Terms such as "treating" or "treatment" or "to treat" refer to (a) therapeutic measures that cure, slow, attenuate, alleviate the symptoms of, and / or stop the progression of a diagnosed pathological condition or disorder, and (b) prophylactic or preventative measures that prevent and / or slow the development of a target disease or condition. Thus, subjects in need of treatment include those already suffering from the cancer, disorder, or disease; those at risk of suffering from the cancer or condition; and those in need of prevention of the infection or condition. Using well-known medical and diagnostic techniques, a subject is identified as "suffering from or at risk of suffering from" sepsis, an infectious disease, an immune system disorder, a metabolic disorder (e.g., diabetes), a hyperproliferative disease, or another disease or disorder mentioned herein. In certain embodiments, if the subject exhibits, for example, all, partial or temporary improvement or elimination of symptoms associated with the disease or condition (e.g., cancer and arthritis such as rheumatoid arthritis), then the subject is successfully "treated" according to the methods provided herein. In specific embodiments, the term "treating" or "treatment" or "to treat" refers to an improvement in at least one measurable physical parameter of a proliferative disorder (such as the growth of a tumor), which improvement is not necessarily recognizable by the patient. In other embodiments, the term "treating" or "treatment" or "to treat" refers to the inhibition of the progression of a proliferative disorder, which can be achieved physically, for example, by the stabilization of recognizable symptoms, physiologically, for example, by the stabilization of physical parameters, or by both. In other embodiments, the term "treating" or "treatment" or "to treat" refers to a reduction or stabilization of tumor size, tumor cell proliferation or survival, or cancer cell counts. Treatment can be performed with provided pharmaceutical compositions disclosed herein (eg, liposomal trans-crocetate) alone or in combination with additional therapeutic agents.

[0214] "Subject" and "patient" and "animal" are used interchangeably and refer to mammals (such as human patients and non-human primates) and experimental animals (such as rabbits, rats and mice), as well as other animals. Animals include all vertebrates, for example, mammals and non-mammals, such as chickens, amphibians and reptiles. "Mammal" as used herein refers to any member of the class Mammalia, including but not limited to humans and non-human primates, such as chimpanzees and other apes and monkey species; livestock, such as cattle, sheep, pigs, goats and horses; domestic mammals, such as dogs and cats; experimental animals, including rodents, such as mice, rats and guinea pigs, etc., and other members of the class Mammalia known in the art. In a specific embodiment, the patient is a human.

[0215] The term "pharmaceutically acceptable carrier" refers to a component of a pharmaceutical formulation other than the active ingredient that is non-toxic to a subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, carriers, excipients, stabilizers, diluents, or preservatives. Pharmaceutically acceptable carriers may include, for example, one or more compatible solid or liquid fillers, diluents, or encapsulating materials suitable for administration to a human or other subject.

[0216] "Therapeutic agent": In some embodiments, the liposome compositions and liposome formulations provided include liposomes that encapsulate or otherwise associate with one or more therapeutic agents, the one or more therapeutic agents being present anywhere in, on, or around the liposomes. For example, the therapeutic agent can be embedded in the lipid bilayer of the liposome, encapsulated in the internal phase of the liposome, or tethered to the exterior of the liposome. The therapeutic agent or agents used in accordance with the disclosed compositions and methods may include any agent intended to treat a condition in a subject. Examples of therapeutic agents that may be suitable for use in accordance with the disclosed methods include vitamin C, thiamine, hydrocortisone or another corticosteroid (e.g., glucocorticoids such as cortisone, ethamethasoneb, prednisone, prednisolone, triamcinolone, dexamethasone, and methylprednisolone; and mineralocorticoids such as fludrocortisonel), astaxanthin, abscisic acid, vitamin A, angiotensin II (e.g., GIAPREZA TM ), tissue plasminogen activator (tPA), antimicrobial agents (e.g., antibiotics), and anti-inflammatory drugs.

[0217] Additional examples of therapeutic agents that may be suitable for use in accordance with the disclosed methods include, without limitation, anti-restenotic agents, pro- or anti-proliferative agents, anti-neoplastic agents, anti-mitotic agents, anti-platelet agents, anti-coagulants, anti-fibrin agents, anti-coagulants, cytostatic agents, antibiotics and other anti-infective agents, anti-enzymatic agents, anti-metabolites, angiogenic agents, cytoprotective agents, angiotensin converting enzyme (ACE) inhibitors, angiotensin II receptor antagonists, and / or cardioprotective agents. In general, any therapeutic agent known in the art may be used, including, but not limited to, those listed in the United States Pharmacopeia (USP); Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10th Edition, McGraw Hill, 2001; Katzung, ed., Basic and Clinical Pharmacology, McGraw-Hill / Appleton & Lange, 8th Edition, September 21, 2000; Physician's Desk Reference (Thomson Publishing); and / or The Merck Manual of Diagnosis and Therapy, 18th Edition, 2006, Beers and Berkow, ed., Merck Publishing Group; or, with respect to animals, The Merck Veterinary Manual, 9th Edition, Kahn, ed., Merck Publishing Group, 2005; all of which are incorporated herein by reference and are used herein to refer to agents or derivatives thereof that can interact with hyperproliferative cells, such as cancer cells or immune cells, thereby reducing the proliferative state of the cells and / or killing the cells. Examples of therapeutic agents include, but are not limited to, chemotherapeutic agents, cytotoxic agents, platinum-based agents (e.g., cisplatin, carboplatin, oxaliplatin), taxanes (e.g., Taxol), etoposide, alkylating agents (e.g., cyclophosphamide, ifosfamide), metabolic antagonists (e.g., methotrexate (MTX), 5-fluorouracil, gemcitabine, pemetrexed or derivatives thereof), antitumor antibiotics (e.g., mitomycin, doxorubicin), plant-derived antitumor agents (e.g., vincristine, vindesine, Taxol). Such agents may further include, but are not limited to, the anticancer agents trimetrexate, temozolomide, TM Raltidracet TM , S-(4-nitrobenzyl)-6-thioinosine (NBMPR), 6-benzylguanidine (6-BG), bischloronitrosourea (BCNU) and camptothecin TM, or a therapeutic derivative of any one of them. "Therapeutic agent" also refers to the salt, acid and free base forms of the above agents.

[0218] The term "pharmaceutically acceptable carrier" refers to a component of a pharmaceutical formulation other than the active ingredient that is non-toxic to a subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, carriers, excipients, stabilizers, diluents, or preservatives. Pharmaceutically acceptable carriers may include, for example, one or more compatible solid or liquid fillers, diluents, or encapsulating materials suitable for administration to a human or other subject.

[0219] The term "kit" refers to a set of two or more components necessary to employ the methods and compositions provided herein. Kit components may include, but are not limited to, liposome compositions and liposome formulations disclosed herein, reagents, buffers, containers and / or equipment. The phrase "separate storage" refers to a storage method for liposomes that prevents a first population of liposomes from contacting another population of liposomes.

[0220] The term "radiosensitizer" refers to a compound that makes tumor cells more sensitive to radiation therapy. Examples of radiosensitizers include misonidazole, metronidazole, tirapazamine, and trans-crocetin.

[0221] Pharmaceutical composition

[0222] The provided pharmaceutical compositions can be prepared in various ways using standard synthetic methods and procedures known to those skilled in the art or obvious to those skilled in the art based on the teachings herein, using commercially available starting materials, compounds known in the literature, or from readily prepared intermediates. Standard synthetic methods and procedures for the preparation of organic molecules and for functional group transformations and manipulations can be obtained from the relevant scientific literature or from standard textbooks in the field. Although not limited to any one or a few sources, classic books such as Smith et al., March's Advanced Organic Chemistry. Reactions, Mechanisms, and Structure, 5th edition, John Wiley & Sons: New York, 2001; Greene, TW, Wuts, PGM, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons: New York, 1999; R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) (each incorporated herein by reference) are useful and recognized reference textbooks on organic synthesis known to those skilled in the art. The following descriptions of synthetic methods are intended to illustrate, but not to limit, general procedures for the preparation of compounds of the present disclosure.

[0223] In some embodiments, the present disclosure provides a new class of multivalent ionizable carotenoid (eg, trans-carotenoid) salts.

[0224] In some embodiments, the present disclosure provides a pharmaceutical composition comprising an ionizable carotenoid having the formula:

[0225] Polyene carotenoids-Q, wherein

[0226] The polyene carotenoids include

[0227] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds,

[0228] (b) a methyl or lower alkyl (C2-C3) substituent, and

[0229] (c) 1, 2, 3 or more ionizable groups; and

[0230] Q is a multivalent counter ion.

[0231] In some embodiments, the polyene carotenoid comprises an all-trans conjugated double bond. In some embodiments, the polyene carotenoid comprises 6-9 conjugated double bonds. In a specific embodiment, the polyene carotenoid comprises 7 conjugated double bonds. The polyene carotenoid may be naturally occurring or synthetic. In some embodiments, the polyene carotenoid is naturally occurring. In other embodiments, the polyene carotenoid is synthetic. The one or more ionizable groups may be anionic and / or cationic. In some embodiments, the polyene carotenoid-Q comprises two or more identical ionizable groups. In some embodiments, the polyene carotenoid comprises two or more different ionizable groups. In some embodiments, the polyene carotenoid comprises one or more anionic ionizable groups. In some embodiments, the polyene carotenoid comprises at least one ionizable group selected from the following: carboxyl, sulfonate, sulfate, phosphonate or phosphate and hydroxamate moieties. In other embodiments, the polyene carotenoid comprises one or more cationic ionizable groups (e.g., primary, secondary or tertiary amine groups, quaternary ammonium groups, choline groups, guanidine groups or imidazolyl groups). In specific embodiments, the polyene carotenoid comprises one or more cationic ionizable groups, and the pharmaceutical composition is substantially free of nucleic acids.

[0232] In some embodiments, Q is a multivalent cationic counterion. In some embodiments, Q is a multivalent metal cation. In further embodiments, Q is a multivalent transition metal cation. In some embodiments, Q is a divalent counterion. In some embodiments, Q is a divalent cationic counterion. In some embodiments, Q is a divalent metal cation. In further embodiments, Q is a divalent transition metal cation. In some embodiments, Q is selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ In some embodiments, Q is Ca 2+ or Mg 2+ In some embodiments, Q is Ca 2+ In further embodiments, the polyene carotenoid-Q is trans-calcium crocetin (CTC). In some embodiments, Q is Mg 2+In further embodiments, the polyene carotenoid-Q is magnesium trans-crocetin (MTC). In other embodiments, Q is a trivalent cationic counterion, such as Fe 3+ In other embodiments, Q is a multivalent organic counterion. In some embodiments, Q is a divalent organic cation. In some embodiments, Q is a divalent organic cation, such as a protonated diamine. Also provided herein are liposomes comprising a polyene carotenoid-Q composition and pharmaceutical compositions (e.g., liposome compositions) comprising the liposomes.

[0233] In some embodiments, the present disclosure provides a pharmaceutical composition comprising an ionizable carotenoid having the formula:

[0234] Q-R1-polyene carotenoid-R2-Q, wherein

[0235] R1-polyene carotenoids-R2 includes

[0236] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds,

[0237] (b) a methyl or lower alkyl (C2-C3) substituent, and

[0238] (c) 1, 2, 3 or more ionizable groups;

[0239] R1 and R2 are ionizable groups; and

[0240] Q is a multivalent counter ion.

[0241] In some embodiments, the Q-R1-polyene carotenoid-R2 comprises an all-trans conjugated double bond. In a specific embodiment, the Q-R1-polyene carotenoid-R2 comprises 6-9 conjugated double bonds. The Q-R1-polyene carotenoid-R2 can be naturally occurring or synthetic. In some embodiments, the Q-R1-polyene carotenoid-R2 is naturally occurring. In other embodiments, the Q-R1-polyene carotenoid-R2 is synthetic. In some embodiments, R1 and R2 are the same ionizable groups. In other embodiments, R1 and R2 are different ionizable groups. In some embodiments, R1 and R2 are the same cationic ionizable groups (e.g., primary, secondary or tertiary amine groups, quaternary ammonium groups, choline groups, guanidine groups and imidazolyl groups). In other embodiments, R1 and R2 are different cationic groups. In some embodiments, R1 and R2 are the same anionic ionizable groups (e.g., carboxyl, sulfonate, sulfate, phosphonate, phosphate, and hydroxamate). In other embodiments, R1 and R2 are different anionic groups. In some embodiments, R1 is a cationic ionizable group or an anionic ionizable group, and R2 is an anionic ionizable group or a cationic group accordingly. In some embodiments, the polyene carotenoid comprises at least one ionizable group selected from the following: carboxyl, sulfonate, sulfate, phosphonate, or phosphate and hydroxamate moieties. In some embodiments, R1 and / or R2 are at least one ionizable group selected from the following: carboxyl, sulfonate, sulfate, phosphonate, or phosphate and hydroxamate moieties. In some embodiments, R2 is at least one ionizable group selected from the following: carboxyl, sulfonate, sulfate, phosphonate or phosphate and hydroxamate moieties. In other embodiments, R1 and / or R2 are cationic ionizable groups (e.g., primary, secondary or tertiary amine groups, quaternary ammonium groups, choline groups, guanidine groups or imidazolyl groups). In specific embodiments, R1 is a cationic ionizable group, and the pharmaceutical composition is substantially free of nucleic acids.

[0242] In some embodiments, Q is a multivalent cationic counterion. In some embodiments, Q is a multivalent metal cation. In further embodiments, Q is a multivalent transition metal counterion. In some embodiments, Q is a divalent counterion. In some embodiments, Q is a divalent cationic counterion. In further embodiments, Q is a divalent metal cation. In some embodiments, Q is selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ In a further embodiment, Q is Ca2+ or Mg 2+ In a further embodiment, Q is Ca 2+ In some embodiments, Q is Mg 2+ In other embodiments, Q is a trivalent cationic counterion such as Fe 3+ In other embodiments, Q is a multivalent organic cation. In further embodiments, Q is a divalent organic cation, such as a protonated diamine. Also provided herein are liposomes comprising the R1-polyene carotenoid-R2 composition and pharmaceutical compositions (e.g., liposome compositions) comprising the liposomes.

[0243] In some embodiments, the present disclosure provides a pharmaceutical composition comprising an ionizable di-α,ω-carotenoid having the formula:

[0244] Q-R1-polyene carotenoid-R1-Q, wherein

[0245] R1-Polyene Carotenoids - R1-Q contains

[0246] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds,

[0247] (b) a methyl or lower alkyl (C2-C3) substituent, and

[0248] (c) 1, 2, 3 or more ionizable groups; and

[0249] R1 is an ionizable group; and

[0250] Q is a multivalent counter ion.

[0251] In some embodiments, the di-α, ω-carotenoid comprises an all-trans conjugated double bond. In some embodiments, the di-α, ω-carotenoid comprises 6-9 conjugated double bonds. In a specific embodiment, the di-α, ω-carotenoid comprises 7 conjugated double bonds. The di-α, ω-carotenoid may be naturally occurring or synthetic. In some embodiments, the di-α, ω-carotenoid is naturally occurring. In other embodiments, the di-α, ω-carotenoid is synthetic. In some embodiments, R1 is an anionic ionizable group. In some embodiments, the di-α, ω-carotenoid comprises an ionizable group selected from the following: carboxyl, sulfonate, sulfate, phosphonate, phosphate and hydroxamate moieties. In other embodiments, R1 is a cationic ionizable group (e.g., primary, secondary or tertiary amine, quaternary ammonium, choline, guanidine or imidazole). In certain embodiments, R1 is a cationic ionizable group, and the pharmaceutical composition is substantially free of nucleic acids.

[0252] In some embodiments, Q is a multivalent cationic counterion. In further embodiments, Q is a multivalent metal cation. In some embodiments, Q is a multivalent transition metal counterion. In some embodiments, Q is a divalent counterion. In some embodiments, Q is a divalent cationic counterion. In further embodiments, Q is a divalent metal cation. In some embodiments, Q is selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ In a further embodiment, Q is Ca 2+ or Mg 2+ In some embodiments, Q is Ca 2+ In some embodiments, Q is Mg 2+ In other embodiments, Q is a trivalent cationic counterion such as Fe 3+ In some embodiments, Q is a multivalent organic cation. In further embodiments, Q is a divalent organic cation, such as a protonated diamine or a protonated polyamine. Also provided herein are liposomes comprising the R1-polyene carotenoid-R1 composition and pharmaceutical compositions (e.g., liposome compositions) comprising the liposomes.

[0253] In some embodiments, the present disclosure provides a pharmaceutical composition comprising an ionizable di-α,ω-carotenoid having the formula: R1-polyene carotenoid-R1, wherein,

[0254] The di-α,ω-carotenoids contain:

[0255] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds, and

[0256] (b) 1, 2, 3 or more ionizable groups; and

[0257] The di-α,ω-carotenoid is optionally substituted with 1 to n methyl or lower C1-C3 alkyl substituents, wherein n=1 to 4; and

[0258] R1 is a polar group and / or a monocyclic functional group.

[0259] In some embodiments, the bis-α,ω-carotenoid comprises an all-trans conjugated double bond. In some embodiments, the bis-α,ω-carotenoid comprises 6-9 conjugated double bonds. In a specific embodiment, the bis-α,ω-carotenoid comprises 7 conjugated double bonds. The bis-α,ω-carotenoid may be naturally occurring or synthetic. In some embodiments, the bis-α,ω-carotenoid is naturally occurring. In other embodiments, the bis-α,ω-carotenoid is synthetic. In some embodiments, R1 is a polar group. In some embodiments, R1 is a monocyclic functional group. In some embodiments, R1 is a polar group and a monocyclic functional group. In some embodiments, the bis-α,ω-carotenoid comprises a monocyclic and / or polar functional group selected from the functional groups present in astaxanthin, lutein, lutein and zeaxanthin. In some embodiments, the bi-α,ω-carotenoid is selected from astaxanthin, lutein, lutein, and zeaxanthin (eg, as shown below).

[0260]

[0261] Also provided herein are liposomes comprising the bi-α,ω-carotenoid composition and pharmaceutical compositions (eg, liposome compositions) comprising the liposomes.

[0262] In some embodiments, the pharmaceutical composition comprises trans-crocetin having the formula: Q-trans-crocetin-Q

[0263]

[0264] in,

[0265] Q is a multivalent cationic counterion.

[0266] In some embodiments, Q is a multivalent metal cation. In further embodiments, Q is a multivalent transition metal cation. In some embodiments, Q is a divalent cationic counterion. In further embodiments, Q is a divalent metal cation. In some embodiments, Q is selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ In a further embodiment, Q is Ca 2+ or Mg 2+ In some embodiments, Q is Ca 2+ In some embodiments, Q is Mg 2+ In other embodiments, Q is a trivalent cationic counterion such as Fe 3+In some embodiments, Q is a multivalent organic cation. In further embodiments, Q is a divalent organic cation, such as a protonated diamine. Also provided herein are liposomes comprising the trans-crocetin composition and pharmaceutical compositions (e.g., liposome compositions) comprising the liposomes.

[0267] In some embodiments, the present disclosure provides a pharmaceutical composition comprising trans-calcium crocetin (CTC). The CTC may exist in linear and / or cyclic forms (as shown below).

[0268]

[0269] Also provided herein are liposomes comprising the CTC composition and pharmaceutical compositions (eg, liposome compositions) comprising the liposomes.

[0270] In some embodiments, the present disclosure provides a pharmaceutical composition comprising trans-magnesium crocetin (MTC). The MTC can exist in linear and / or cyclic forms (as shown below).

[0271]

[0272] Also provided herein are liposomes comprising the MTC compositions and pharmaceutical compositions (eg, liposome compositions) comprising the liposomes.

[0273] In some embodiments, the present disclosure provides a pharmaceutical composition comprising trans-norbixin having the formula: Q-norbixin-Q

[0274]

[0275] in,

[0276] Q is a multivalent cationic counterion.

[0277] In some embodiments, Q is a multivalent cationic counterion. In some embodiments, Q is a multivalent metal cation. In further embodiments, Q is a multivalent transition metal cation. In some embodiments, Q is a divalent cationic counterion. In further embodiments, Q is a divalent metal cation. In some embodiments, Q is selected from Ca 2+ Mg 2 + 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ In a further embodiment, Q is Ca 2+ or Mg 2+ In some embodiments, Q is Ca 2+In some embodiments, Q is Mg 2+ In other embodiments, Q is a trivalent cationic counterion such as Fe 3+ In some embodiments, Q is a multivalent organic cation. In further embodiments, Q is a divalent organic cation, such as a protonated diamine. Also provided herein are liposomes comprising the trans-norbixin composition and pharmaceutical compositions (e.g., liposome compositions) comprising the liposomes.

[0278] In some embodiments, the present disclosure provides a pharmaceutical composition comprising trans-calcium norbixin (CTN). The CTN may exist in linear and / or cyclic forms (as shown below),

[0279]

[0280] Also provided herein are liposomes comprising CTN and pharmaceutical compositions (eg, liposome compositions) comprising the liposomes.

[0281] In some embodiments, the present disclosure provides a pharmaceutical composition comprising trans-magnesium norbixin (MTN). The MTN can exist in linear and / or cyclic forms (as shown below),

[0282]

[0283] Also provided herein are liposomes comprising MTN and pharmaceutical compositions (eg, liposome compositions) comprising the liposomes.

[0284] The lipids and other components of the liposomes contained in the liposome composition can be any lipid, lipid combination and ratio known in the art, or a combination of lipids and other liposome components and their corresponding ratios. However, it will be appreciated by those skilled in the art that the liposome encapsulation of any particular drug (such as, but not limited to, the carotenoid composition discussed herein) may involve a large number of routine experiments to obtain useful functional liposome preparations. In general, the liposomes provided can have any liposome structure, for example, a structure having an internal space isolated from an external medium by one or more lipid bilayers, or a microcapsule having a semipermeable membrane and a lipophilic core portion (wherein the membrane isolates the interior). The lipid bilayer can be any arrangement of amphiphilic molecules, characterized in that the amphiphilic molecules are characterized by a hydrophilic portion (hydrophilic portion) and a hydrophobic portion (hydrophobic portion). The amphiphilic molecules in the bilayer are generally arranged into a two-dimensional sheet, wherein the hydrophobic portion is oriented inwardly of the sheet, and the hydrophilic portion is oriented outwardly. The amphiphilic molecules forming the provided liposomes can be any known or later discovered amphiphilic molecules, for example, lipids or biocompatible lipids of synthetic or natural origin. The liposomes can also be formed from amphiphilic polymers and surfactants, for example, polymerosomes and niosomes. For the purposes of this disclosure, but not limitation, these liposome-forming materials are also referred to as "lipids."

[0285] The liposome composition preparation provided herein can be in liquid or dry form, such as dry powder or dry cake. The dry powder or dry cake may be subjected to primary drying under, for example, freeze-drying conditions, or optionally, the dry cake or dry powder may only be subjected to primary drying, or primary drying and secondary drying. In dry form, the powder or cake may, for example, have a moisture content between 1% and 6%, for example, such as a moisture content between 2% and 5% or a moisture content between 2% and 4%. An example method of drying is freeze drying (also referred to as freeze drying or cryogenic drying). Any composition and method disclosed herein may include liposomes, freeze-dried liposomes, or liposomes reconstructed from freeze-dried liposomes. In some embodiments, the composition and method include one or more freeze-dried protective agents or cryoprotectants. These protective agents are typically polyhydroxy compounds, such as sugars (monosaccharides, disaccharides, and polysaccharides), polyols and derivatives thereof, glycerol or polyethylene glycol, trehalose, maltose, sucrose, glucose, lactose, dextran, glycerol, or aminoglycosides. In further embodiments, the lyoprotectant or cryoprotectant comprises at most 10% or at most 20% of the solution outside the liposomes, inside the liposomes, or both.

[0286] The characteristics of liposomes are affected by the lipid properties used to prepare liposomes. Various lipids have been used to prepare liposomes. These lipids include cationic, anionic and neutral lipids. In some embodiments, the liposomes comprising carotenoid compositions (e.g., CTC and MTC) are anionic or neutral. In other embodiments, the liposomes provided are cationic. The determination of charge (e.g., anionic, neutral or cationic) can be routinely determined by measuring the zeta potential of the liposomes. The zeta potential of the liposome can be a positive number, zero or negative number. In some embodiments, the zeta potential of the liposome is -150 to 150mV, or -50 to 50mV, or any range therebetween. In some embodiments, the zeta potential of the liposome is less than or equal to zero. In some embodiments, the zeta potential of the liposome is -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween. In other embodiments, the zeta potential of the liposome is greater than zero. In some embodiments, the liposomes have a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween.

[0287] In some embodiments, cationic lipids are used to prepare cationic liposomes, which are commonly used as gene transfection agents. The positive charge on the cationic liposome can interact with the negative charge on the cell surface. After the cationic liposome binds to the cell, the liposome is transported within the cell by phagocytosis.

[0288] In some preferred embodiments, neutral to anionic liposomes are used. In preferred embodiments, anionic liposomes are used. The use of, for example, a mixture of neutral lipids (such as HSPC) and anionic lipids (such as PEG-DSPE) results in the formation of anionic liposomes, which are less likely to non-specifically bind to normal cells. Specific binding to tumor cells can be achieved by using tumor targeting antibodies, such as, for example, folate receptor antibodies including, for example, folate receptor alpha antibodies, folate receptor beta antibodies and / or folate receptor delta antibodies.

[0289] As an example, at least one (or some) lipid is an amphipathic lipid, which is defined as having a hydrophilic and hydrophobic portion (generally a hydrophilic head and a hydrophobic tail). The hydrophobic portion is generally toward the hydrophobic phase (for example, inside the bilayer), and the hydrophilic portion is generally toward the aqueous phase (for example, outside the bilayer). The hydrophilic portion can include polar or charged groups, such as carbohydrates, phosphate radicals, carboxyls, sulfate radicals, amino, sulfhydryls, nitro, hydroxyls and other similar groups. The hydrophobic portion can include non-polar groups, including but not limited to long-chain saturated and unsaturated aliphatic hydrocarbon groups and groups substituted by one or more aromatic, cycloaliphatic or heterocyclic groups. The example of amphipathic compounds includes but is not limited to phospholipids, amino lipids and sphingolipids.

[0290] Typically, for example, the lipid is a phospholipid. Phospholipids include, but are not limited to, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, etc. It should be understood that other lipid membrane components, such as cholesterol, sphingomyelin, and cardiolipin, can be used.

[0291] The lipid comprising liposome provided herein can be anionic and neutral (including zwitterionic and polar) lipids, including anionic and neutral phospholipids. Neutral lipids exist in the form of uncharged or neutral zwitterionics at selected pH values. At physiological pH, such lipids include, for example, dioleoylphosphatidylglycerol (DOPG), diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebrosides and diacylglycerols. The example of zwitterionic lipids includes, but is not limited to, dioleoylphosphatidylcholine (DOPC), dimyristoylphosphatidylcholine (DMPC) and dioleoylphosphatidylserine (DOPS). Anionic lipids are negatively charged at physiological pH. These lipids include, but are not limited to, phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N-dodecanoylphosphatidylethanolamine, N-succinylphosphatidylethanolamine, N-glutarylphosphatidylethanolamine, lysylphosphatidylglycerol, palmitoyloleoylphosphatidylglycerol (POPG), and other anionic modifying groups conjugated to neutral lipids.

[0292] Anionic lipids and neutral lipids are collectively referred to as non-cationic lipids in this article. Such lipids can contain phosphorus, but they are not limited thereto. The example of non-cationic lipids includes lecithin, lysolecithin, phosphatidylethanolamine, lysophosphatidylethanolamine, dioleoylphosphatidylethanolamine (DOPE), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), distearoylphosphatidyl 1-ethanolamine (DSPE), palmitoyloleoylphosphatidylethanolamine (POPE), palmitoyloleoylphosphatidylcholine (POPC), egg phosphatidylcholine (EPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC ), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), palmitoyloleoylphosphatidylglycerol (POPG), 16-0-monomethyl PE, 16-0-dimethyl PE, 18-1-trans-PE, palmitoyloleoylphosphatidylethanolamine (POPE), 1-stearoyl-2-oleoylphosphatidylethanolamine (SOPE), phosphatidylserine, phosphatidylinositol, sphingomyelin, cephalin, cardiolipin, phosphatidic acid, cerebroside, dicetyl phosphate and cholesterol.

[0293] The liposomes can be assembled using any liposome assembly method using liposome components known in the art (also referred to as liposome components). Liposome components include, for example, lipids such as DSPE, HSPC, cholesterol, and derivatives of these components. Other suitable lipids can be commercially available, for example, from Avanti Polar Lipids, Inc. (Alabaster, Alabama, USA). A partial list of available negatively charged or neutrally charged lipids suitable for preparing anionic liposomes can be, for example, at least one of the following: DLPC, DMPC, DPPC, DSPC, DOPC, DMPE, DPPE, DOPE, DMPA Na, DPPA Na, DOPA Na, DMPG Na, DPPG Na, DOPG Na, DMPS Na, DPPS Na, DOPS Na, DOPE-glutaryl (Na) 2, tetramyristoyl cardiolipin (Na) 2, DSPE-mPEG-2000 Na, DSPE-mPEG-5000 Na, and DSPE-maleimide PEG-2000 Na.

[0294] In some embodiments, the compositions provided are formulated with liposomes comprising cationic lipids. In one embodiment, the cationic lipids are selected from, but not limited to, International Application Publication Nos. WO2012 / 040184, WO2011 / 153120, WO2011 / 149733, WO2011 / 090965, WO2011 / 043913, WO2011 / 022460, WO2012 / 061259, WO2012 / 054365, WO2012 / 04 Cationic lipids described in WO2010 / 04638, WO2010 / 080724, WO2010 / 21865 and WO2008 / 103276, U.S. Patent Nos. 7,893,302, 7,404,969 and 8,283,333, and U.S. Application Publication Nos. US20100036115 and US20120202871; each of which is incorporated herein by reference in its entirety. In another embodiment, the cationic lipid may be selected from, but is not limited to, Formula A described in International Application Publication Nos. WO2012 / 040184, WO2011 / 153120, WO201 / 1149733, WO2011 / 090965, WO2011 / 043913, WO2011 / 022460, WO2012 / 061259, WO2012 / 054365 and WO2012 / 044638; each of which is incorporated herein by reference in its entirety. In yet another embodiment, the cationic lipid may be selected from, but is not limited to, Formula CLI-CLXXIX of International Publication No. WO2008103276, Formula CLI-CLXXIX of U.S. Pat. No. 7,893,302, Formula CLI-CLXXXXII of U.S. Pat. No. 7,404,969, and Formula I-VI of U.S. Patent Publication No. US20100036115; each of which is herein incorporated by reference in its entirety. As non-limiting examples, the cationic lipid may be selected from (20Z, 23Z)-N,N-dimethylnonacoustic-20,23-dien-10-amine, (17Z, 20Z)-N,N-dimethylhexacosa-17,20-dien-9-amine, (1Z, 19Z)-N5N-dimethylpentacoustic-16,19-dien-8-amine, (13Z, 16Z)-N,N-dimethyldocosa-13,16-dien-5-amine, -amine, (12Z,15Z)-N,N-dimethylhexene-12,15-dien-4-amine, (14Z,17Z)-N,N-dimethyltricosac-14,17-dien-6-amine, (15Z,18Z)-N,N-dimethyltetracosa-15,18-dien-7-amine, (18Z,21Z)-N,N-dimethylheptacosac-18,21-dien-10-amine, (15Z,18Z)-N,N-dimethylheptacosa-18,21-dien-10-amine,N-dimethyltetracosa-15,18-dien-5-amine, (14Z,17Z)-N,N-dimethyltricosa-14,17-dien-4-amine, (19Z,22Z)-N,N-dimethyloctacosa-19,22-dien-9-amine, (18Z,21Z)-N,N-dimethylheptacosa-18,21-dien-8-amine, (17Z,20Z)-N,N-dimethylhexacosa-17,20-dien-7-amine, (16Z,19Z)-N,N-dimethylpentacosa-16,19-dien-6-amine, (22Z,25Z)-N,N-dimethylhetriconadecosa-22,25-dien-10-amine, (21Z,24Z)-N,N-dimethyl 2-[(11Z,14Z)-1-nonyleicos-11,14-dien-1-yl]pyrrolidine, (20Z)-N,N-dimethylheptacos-18-en-10-amine, (17Z)-N,N-dimethylhexacos-17-en-9-amine, (19Z,22Z)-N,N-dimethyloctacos-19,22-dien-7-amine, N,N-dimethylheptacos-10-amine, (20Z,23Z)-N-ethyl-N-methylnonacosa-20,23-dien-10-amine, 1-[(11Z,14Z)-1-nonyleicos-11,14-dien-1-yl]pyrrolidine, (20Z)-N,N-dimethylheptacos-20-en-10-amine, (15Z)-N,N-dimethylheptacos-15 -en-10-amine, (14Z)-N,N-dimethylnonacos-14-en-10-amine, (17Z)-N,N-dimethylnonacos-17-en-10-amine, (24Z)-N,N-dimethyltricosa-24-en-10-amine, (20Z)-N,N-dimethylnonacos-20-en-10-amine, (22Z)-N,N-dimethyltricosa-22-en-10-amine, (16Z)-N,N-dimethylpentacos-16-en-8-amine, (12Z,15Z)-N,N-dimethyl-2-nonylhexona-12,15-dien-1-amine, (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1 -amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]heptadecan-8-amine, 1-[(1S,2R)-2-hexylcyclopropyl]-N,N-dimethylnonadecan-10-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]nonadecan-10-amine, N,N-dimethyl-21-[R1S,2R)-2-octylcyclopropyl]heneicosane-10-amine, N,N-dimethyl-1-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]methyl}cyclopropyl]nonadecan-10-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]hexadecan-8-amine, N,N-dimethyl-[(1R,2S)-2-undecyl-cyclopropyl]tetradec-5-amine, N,N-dimethyl-3-{7-[(1S,2R)-2-octylcyclopropyl]heptyl}dodecan-1-amine, 1-[(1R,2S)-2-heptylcyclopropyl]-N,N-dimethyloctadecan-9-amine, 1-[(1S,2R)-2-decylcyclopropyl]-N,N-dimethylpentadecan-6-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]pentadecan-8-amine, RN,N-dimethyl-1-[(9Z,12Z)-octadecan-9,12-dien-1-yloxy]-3-(octyloxy)propan-2-amine, SN,N-dimethyl-1-[(9Z,12Z)-octadecan- 9,12-dien-1-yloxy]-3-(octyloxy)propan-2-amine, 1-{2-[(9Z,12Z)-octadec-9,12-dien-1-yloxy]-1-[(octyloxy)methyl]ethyl}pyrrolidine, (2S)-N,N-dimethyl-1-[(9Z,12Z)-octadec-9,12-dien-1-yloxy]-3-[(5Z-)-oct-5-en-1-yloxy]propan-2-amine, 1-{2-[(9Z,12Z)-octadec-9,12-dien-1-yloxy]-1-[(octyloxy)methyl]ethyl}azetidine, (2S)-1-(hexyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadec-9,12-dien-1-yloxy] oxy]propan-2-amine, (2S)-1-(heptyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadec-9,12-dien-1-yloxy]propan-2-amine, N,N-dimethyl-1-(nonyloxy)-3-[(9Z,12Z)-octadec-9,12-dien-1-yloxy]propan-2-amine, N,N-dimethyl-1-[(9Z)-octadec-9-en-1-yloxy]-3-(octyloxy)propan-2-amine; (2S)-N,N-dimethyl-1-[(6Z,9Z,12Z)-octadec-6,9,12-trien-1-yloxy]-3-(octyloxy)propan-2-amine, (2S)-1-[(11Z,14Z)-eicosyl-11,1 4-dien-1-yloxy]-N,N-dimethyl-3-(pentyloxy)propan-2-amine, (2S)-1-(hexyloxy)-3-[(11Z,14Z)-eicosyl-11,14-dien-1-yloxy]-N,N-dimethylpropan-2-amine, 1-[(11Z,14Z)-eicosyl-11,14-dien-1-yloxy]-N,N-dimethyl1-3-(octyloxy)propan-2-amine, 1-[(13Z,16Z)-eicosyl-13,16-dien-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, (2S)-1-[(13Z,16Z)-eicosyl-13,16-dien-1-yloxy]-3-(hexyloxy)-N,N-dimethyl-propan-2-amine, (2S)-1-[(13Z)-docosa-13-en-1-yloxy]-3-(hexyloxy)-N,N-dimethylpropan-2-amine, 1-[(13Z)-docosa-13-en-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, 1-[(9Z)-hexadec-9-en-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, (2R)-N,N-dimethyl-H(1-methyloctyl)oxy]-3-[(9Z,12Z)-octadec-9,12-dien-1-yloxy]propan-2-amine, (2R)-1-[(3, 7-dimethyloctyl)oxy]-N,N-dimethyl-3-R9Z,12Z)-octadec-9,12-dien-1-yloxypropan-2-amine, N,N-dimethyl-1-(octyloxy)-3-({8-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]-methyl}cyclopropyl]octyl}oxy)propan-2-amine, N,N-dimethyl-1-{[-(2-octylcyclopropyl)octyl]oxy}-3-(octyloxy)propan-2-amine and (11E,20Z,23Z)-N,N-dimethylnonacoustic-11,20,2-trien-10-amine or their pharmaceutically acceptable salts or acids or stereoisomers. ,

[0295] In one embodiment, the lipid may be a cleavable lipid, such as those described in International Publication No. WO 2012 / 170889, which is herein incorporated by reference in its entirety.

[0296] The cationic lipids can be conventionally synthesized using methods known in the art (see, e.g., International Publication Nos. WO2012 / 040184, WO2011 / 153120, WO2011 / 149733, WO2011 / 090965, WO201 / 1043913, WO2011 / 022460, WO2012 / 061259, WO2012 / 054365, WO2012 / 044638, WO2010 / 080724, and WO2010 / 21865; each of which is incorporated herein by reference in its entirety).

[0297] The lipid derivative may include at least, for example, one or more steric stabilizers and / or functional groups bonded to the liposome component (preferably covalently bonded), after which the steric stabilizer and / or functional group should be considered as part of the liposome component. Functional groups include groups that can be used to connect the liposome component to another part (such as a protein). Such functional groups include at least maleimide. These steric stabilizers include at least one selected from the group consisting of: polyethylene glycol (PEG); poly-L-lysine (PLL); monosialoganglioside (GM1); poly(vinyl pyrrolidone) (PVP); poly(acrylamide) (PAA); poly(2-methyl-2-oxazoline); poly(2-ethyl-2-oxazoline); phosphatidyl polyglycerol; poly[N-(2-hydroxypropyl) methacrylamide]; amphiphilic poly-N-vinyl pyrrolidone; L-amino acid-based polymers; and polyvinyl alcohol.

[0298] In some embodiments, the carotenoid compositions provided are formulated as lipid-polycation complexes. The formation of the lipid-polycation complexes can be achieved using methods known in the art and / or described in U.S. Publication No. 2012 / 0178702, which is incorporated herein by reference in its entirety. As non-limiting examples, the polycations may include cationic peptides or polypeptides (such as, but not limited to, polylysine, polyornithine, and / or polyarginine), and cationic peptides described in International Publication No. WO2012 / 013326, which is incorporated herein by reference in its entirety. In another embodiment, the carotenoid compositions provided are formulated as lipid-polycation complexes, which further comprise a neutral lipid, such as, but not limited to, cholesterol or dioleoylphosphatidylethanolamine (DOPE).

[0299] Since the components of the liposomes can include any one or more molecules (i.e., chemicals / reagents / proteins) bound thereto, in some embodiments, the components of the provided liposomes include at least a member selected from the following: DSPE, DSPE-PEG, DSPE-maleimide, HSPC; HSPC-PEG; HSPC-maleimide; cholesterol; cholesterol PEG; and cholesterol-maleimide. In some embodiments, the components of the provided liposomes include DSPE, DSPE-PEG, DSPE-maleimide, HSPC, HSPC-PEG; HSPC-maleimide; cholesterol; cholesterol PEG; and cholesterol-maleimide. In a preferred embodiment, the liposome components constituting the liposomes include DSPE, DSPE-FITC, DSPE-maleimide, cholesterol, and HSPC.

[0300] In other embodiments, the liposomes of the liposome compositions provided herein include oxidized phospholipids. In some embodiments, the liposomes include oxidized phospholipids of members selected from phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, phosphatidylcholine and 1-palmitoyl-2-arachidonoyl-sn-glyceryl-2-phosphate. In some embodiments, the phospholipids have unsaturated bonds. In some embodiments, the phospholipids are phospholipids containing arachidonic acid. In other embodiments, the phospholipids are sn-2-oxygenated. In other embodiments, the phospholipids are not fragmented.

[0301] In some embodiments, the liposomes of the disclosed liposome composition contain oxidized 1-palmitoyl-2-arachidonoyl-sn-glyceryl-3-phosphorylcholine (OxPAPC). As used herein, the term "oxPAPC" refers to lipids produced by oxidation of 1-palmitoyl-2-arachidonoyl-sn-glyceryl-3-phosphorylcholine (PAPC), which produces a mixture of oxidized phospholipids containing fragmented or full-length oxygenated sn-2 residues. Characterized oxidative fragmented substances contain five-carbon sn-2 residues with ω-aldehyde or ω-carboxyl groups. Oxidation of arachidonic acid residues also produces phospholipids containing esterified isoprostanes. oxPAPC includes HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC substances, as well as other oxidation products present in oxPAPC. In a further embodiment, the oxPAPC is a phospholipid containing epoxyisoprostanoids. In further embodiments, the oxPAPC is 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glyceryl-3-phosphocholine (5,6-PEIPC), 1-palmitoyl-2-(epoxy-cyclopentenone)-sn-glyceryl-3-phosphocholine (PECPC) and / or 1-palmitoyl-2-(epoxy-isoprostaglandin E2)-sn-glyceryl-4-phosphocholine (PEIPC). In some embodiments, the phospholipid has an unsaturated bond. In some embodiments, the phospholipid is a phospholipid containing arachidonic acid. In other embodiments, the phospholipid is sn-2-oxygenated. In other embodiments, the phospholipid is not fragmented.

[0302] In some embodiments, the liposomes of the disclosed liposome composition comprise lipids selected from the group consisting of 1-palmitoyl-2-glutaryl-sn-glyceryl-3-phosphocholine (PGPC); 1-palmitoyl-2-(9'-oxy-nonanoyl)-sn-glyceryl-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glyceryl-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glyceryl-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glyceryl-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glyceryl-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glyceryl-3-phosphocholine. In further embodiments, the liposomes comprise PGPC.

[0303] In some embodiments, at least one component of the liposome lipid bilayer is functionalized (or reactive). As used herein, a functionalized component is a component comprising a reactive group, which can be used to crosslink reagents and moieties with lipids. If the lipid is functionalized, any liposome formed by it is also functionalized. In some embodiments, the reactive group is a group that will react with a crosslinking agent (or other moieties) to form a crosslink. The reactive group in the liposome lipid bilayer is located on the lipid so that it can contact with a crosslinking agent and crosslink with another moiety (e.g., a steric stabilizer or a targeting moiety) at any position. In some embodiments, the reactive group is located in the head group of a lipid including, for example, a phospholipid. In some embodiments, the reactive group is a maleimide group. Maleimide groups can be crosslinked to each other in the presence of a dithiol crosslinking agent, and the dithiol crosslinking agent includes, but is not limited to, dithiothreitol (DTT).

[0304] It should be understood that the use of other functionalized lipids, other reactive groups and other cross-linking agents other than those described above is further contemplated. In addition to maleimide groups, other examples of reactive groups contemplated include, but are not limited to, other thiol reactive groups, amino groups (such as primary and secondary amines), carboxyl groups, hydroxyl groups, aldehyde groups, alkynyl groups, azido groups, carbonyl groups, haloacetyl groups (e.g., iodoacetyl groups), imido ester groups, N-hydroxysuccinimide esters, sulfhydryl groups and pyridyl disulfide groups.

[0305] Functionalized and non-functionalized lipids are available from a number of commercial sources including Avanti Polar Lipids (Alabaster, AL) and Lipoid LLC (Newark, NJ).

[0306] In some embodiments, the liposomes contain a steric stabilizer that increases the life of the liposomes in circulation. One or more steric stabilizers, such as hydrophilic polymers (polyethylene glycol (PEG)), glycolipids (monosialyl ganglioside (GM1)), etc., occupy the space next to the surface of the liposomes and exclude other macromolecules from this space. As a result, the access and binding of blood plasma opsonins to the surface of the liposomes are hindered, so the interaction of macrophages with such liposomes or any other clearance mechanism is inhibited, and the life of the liposomes in circulation is increased. In some embodiments, the steric stabilizer or steric stabilizer group is PEG or a combination comprising PEG. In a further embodiment, the steric stabilizer is PEG or a combination comprising PEG having a number average molecular weight (Mn) of 200 to 5000 Daltons. The one or more PEGs can have any structure, such as a linear, branched, star or comb structure, and can be commercially available.

[0307] In some embodiments, the liposomes of the provided liposome compositions are pegylated (e.g., pegylated liposome CTC and pegylated liposome MTC). In some embodiments, the pegylated liposomes are water-soluble. That is, the pegylated liposomes are in the form of an aqueous solution.

[0308] The diameter of the liposome provided is not particularly limited. In some embodiments, the liposome has, for example, an average diameter of 20 nm to 500 nm (nanometer), or 20 nm to 200 nm, or any range therebetween. In some embodiments, the liposome has an average diameter of 80 nm to 120 nm, or any range therebetween.

[0309] In some embodiments, the pH of the solution comprising the liposome composition is 2 to 8, or any range therebetween. In some embodiments, the pH of the solution comprising the liposome composition is 5 to 8, or 6 to 7, or any range therebetween. In some embodiments, the pH of the solution comprising the liposome composition is 6 to 7, or any range therebetween. In some embodiments, the pH of the solution comprising the liposome composition is 6 to 7.5, 6.5 to 7.5, 6.7 to 7.5, or 6.3 to 7.0, or any range therebetween.

[0310] In other embodiments, the liposome composition provided includes a buffer. In further embodiments, the buffer is selected from HEPES, citrate or sodium phosphate (e.g., sodium dihydrogen phosphate and / or disodium hydrogen phosphate). In some embodiments, the buffer is HEPES. In some embodiments, the buffer is citrate. In some embodiments, the buffer is sodium phosphate (e.g., sodium dihydrogen phosphate and / or disodium hydrogen phosphate). In some embodiments, the buffer is in 15 to 200mM, or at a concentration of any range therebetween. In further embodiments, the buffer is in 5 to 200mM, 15 to 200, 5 to 100mM, 15 to 100mM, 5 to 50mM, 15 to 50mM, 5 to 25mM, 5 to 20mM, 5 to 15mM, or at a concentration of any range therebetween. In some embodiments, the buffer is HEPES, which is in 5 to 200mM or at a concentration of any range therebetween. In some embodiments, the buffer is citrate at a concentration of 5 to 200 mM or any range therebetween. In some embodiments, the buffer is sodium phosphate at a concentration of 5 to 200 mM or any range therebetween.

[0311] In other embodiments, the liposome composition contains one or more lyoprotectants or cryoprotectants. In some embodiments, the cryoprotectant is mannitol, trehalose, sorbitol or sucrose. In some embodiments, the lyoprotectant and / or cryoprotectant is present in the composition in an amount of 1 to 20%, or 5 to 20% by weight, or any range therebetween.

[0312] In other embodiments, the liposome composition provided comprises a tonicity agent. In some embodiments, the concentration (weight percent) of the tonicity agent is 0.1-20%, 1-20%, 0.5-15%, 1-15%, or 1-50%, or any range therebetween. In some embodiments, the liposome composition comprises sugar (e.g., trehalose, maltose, sucrose, lactose, mannose, mannitol, glycerol, dextrose, fructose, etc.). In further embodiments, the concentration (weight percent) of the sugar is 0.1-20%, 1-20%, 0.5-15%, 1%-15%, or 1-50%, or any range therebetween.

[0313] In some embodiments, the liposome composition provided comprises trehalose. In a further embodiment, the concentration weight percentage of trehalose is 0.1-20%, 1-20%, 0.5-15%, 1%-15%, 5-20%, or 1-50%, or any range therebetween. In a further embodiment, the concentration (weight percentage) of trehalose is 1-15%, or any range therebetween. In another embodiment, the trehalose is present in a trehalose of about 5% to 20% weight percentage, or in any combination of one or more lyophilization protectants or cryoprotectants with a total concentration of 5% to 20%. In some embodiments, the pH value of the liposome composition is 6 to 7.5, 6.5 to 7.5, 6.7 to 7.5, or 6.3 to 7.0, or any range therebetween.

[0314] In some embodiments, the liposome composition comprises dextrose. In some embodiments, the concentration weight percentage of dextrose is 0.1-20%, 1-20%, 0.5-15%, 1-15%, 5-20%, or 1-50%, or any range therebetween. In a specific embodiment, the concentration (weight percentage) of dextrose is 1-20% or any range therebetween. In other embodiments, the dextrose is present in a dextrose of 1 to 20% weight percentage, or is present in a total concentration of 1% to 20%, or 5% to 20%, or any combination of one or more lyophilizing protectants or cryoprotectants of any range therebetween.

[0315] In some embodiments, the present disclosure provides a liposome composition comprising a liposome encapsulating an ionizable carotenoid salt. In some embodiments, the composition comprises a liposome encapsulating an ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the liposome is pegylated. In some embodiments, the liposome is targeted. In some embodiments, the liposome is unpegylated and targeted. In some embodiments, the liposome is unpegylated and non-targeted. In some embodiments, the liposome contains less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, less than 10,000, or less than 5,000 ionizable carotenoid molecules. In some embodiments, the liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of ionizable carotenoid molecules. In other embodiments, the encapsulated ionizable carotenoid is trans-crocetin, trans-norbixin, or any of the liposomes described herein [1]-

[28] and / or Figures 1A-1DIn some embodiments, the liposomes encapsulate a plurality of carotenoids. In further embodiments, the liposomes encapsulate a plurality of ionizable carotenoids (e.g., a combination of trans-crocetin, trans-norbixin, and / or the liposomes described in [1]-

[28] and / or Figures 1A-1D ionizable carotenoids provided in

[0316] In some embodiments, the present disclosure provides a liposome composition comprising a non-PEGylated non-targeted liposome encapsulating an ionizable carotenoid salt. In some embodiments, the liposome encapsulates an ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the encapsulated ionizable carotenoid is trans-crocetin. In some embodiments, the encapsulated ionizable carotenoid is trans-norbixin. In some embodiments, the encapsulated ionizable carotenoid is any of [1]-

[28] and / or Figures 1A-1D In some embodiments, the non-PEGylated non-targeted liposomes contain less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, less than 10,000, or less than 5,000 carotenoid molecules. In some embodiments, the liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of ionizable carotenoid molecules. In further embodiments, the liposomes encapsulate a plurality of carotenoids. In some embodiments, the liposomes encapsulate a plurality of ionizable carotenoids (e.g., a combination of trans-crocetin, trans-norbixin, and / or the liposomes described herein [1]-

[28] and / or Figures 1A-1D One or more ionizable carotenoids provided in ).

[0317] In some embodiments, the present disclosure provides a liposome composition comprising a PEGylated liposome encapsulating an ionizable carotenoid salt. In some embodiments, the liposome encapsulates an ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the PEGylated liposome contains less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, less than 10,000, or less than 5,000 ionizable carotenoid molecules. In some embodiments, the PEGylated liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range of carotenoid molecules therebetween. In some embodiments, the encapsulated ionizable carotenoid is trans-crocetin. In some embodiments, the encapsulated ionizable carotenoid is trans-norbixin. In some embodiments, the encapsulated ionizable carotenoid is described in [1]-

[28] and / or Figures 1A-1D In some embodiments, the PEGylated liposomes encapsulate a plurality of carotenoids. In further embodiments, the liposomes encapsulate a plurality of ionizable carotenoids (e.g., a combination of trans-crocetin, trans-norbixin and / or the compositions described herein [1]-

[28] and / or Figures 1A-1D In some embodiments, the PEGylated liposome is targeted. In some embodiments, the PEGylated liposome is non-targeted.

[0318] In some embodiments, the present disclosure provides a liposome composition comprising a targeted liposome encapsulating an ionizable carotenoid salt. In some embodiments, the liposome encapsulates the ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the targeted liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest. In some embodiments, the targeting moiety is attached to one or both of PEG and the exterior of the liposome. In some embodiments, the targeting moiety is a polypeptide. In further embodiments, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the targeting moiety binds to a surface antigen with an equilibrium dissociation constant (Kd) such as that obtained using The analysis was performed at 50 x 10 -12 Up to 10x 10 -6In a further embodiment, the Kd is determined using a surface plasmon resonance technique in which an antigen containing the epitope is immobilized, a targeting moiety is used as an analyte, and the following conditions are used: 10 mM MES buffer, 0.05% polyoxyethylene sorbitan monolaurate, and 150 mM NaCl, at 37°C. In some embodiments, the targeting moiety comprises a protein or folate conjugate having a specific affinity for one or more folate receptors selected from the group consisting of folate receptor α (FR-α), folate receptor β (FR-β), and folate receptor δ (FRδ). In some embodiments, the targeted liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range therebetween, of targeting moieties. The targeted liposome may be pegylated or not pegylated.

[0319] In some embodiments, the targeted liposome is pegylated and encapsulates an ionizable carotenoid salt. In some embodiments, the liposome encapsulates an ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the targeted pegylated liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200 targeting moieties or any range therebetween. In some embodiments, the targeted pegylated liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween of ionizable carotenoid molecules. In some embodiments, the encapsulated ionizable carotenoid is trans-crocetin. In some embodiments, the encapsulated ionizable carotenoid is trans-norbixin. In some embodiments, the encapsulated ionizable carotenoid is described in [1]-

[28] and / or Figures 1A-1D Carotenoids provided by

[0320] In other embodiments, the targeted liposome is unpegylated and encapsulates an ionizable carotenoid salt. In some embodiments, the liposome encapsulates an ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the targeted unpegylated liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range therebetween, of targeting moieties. In further embodiments, the liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of ionizable carotenoid molecules. In some embodiments, the encapsulated ionizable carotenoid is trans-crocetin. In some embodiments, the encapsulated ionizable carotenoid is trans-norbixin. In some embodiments, the encapsulated ionizable carotenoid is described herein [1]-

[28] and / or Figures 1A-1D In another embodiment, the targeted unPEGylated liposomes encapsulate a plurality of carotenoids. In a further embodiment, the liposomes encapsulate a plurality of ionizable carotenoids (e.g., a combination of trans-crocetin, trans-norbixin and / or the liposomes described herein [1]-

[28] and / or Figures 1A-1D One or more ionizable carotenoids provided in ).

[0321] In some embodiments, the present disclosure provides a PEGylated liposome composition comprising a liposome encapsulating any provided ionizable carotenoid salt, and encapsulating an ionizable carotenoid salt. In some embodiments, the liposome encapsulates the ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the liposome contains Less than 6 million In some embodiments, the PEGylated liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of ionizable carotenoid molecules. In other embodiments, the liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D Two or more of the ionizable carotenoids provided in.

[0322] In further embodiments, the present disclosure provides liposome compositions comprising PEGylated and targeted liposomes encapsulating ionizable carotenoid salts. In some embodiments, the liposomes encapsulate the ionizable carotenoid salts of any one of [1]-

[28] . In some embodiments, the PEGylated and targeted liposomes contain 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range therebetween, of targeting moieties. In some embodiments, the PEGylated and targeted liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of ionizable carotenoid molecules. In further embodiments, the liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D Two or more of the ionizable carotenoids provided in.

[0323] In another embodiment, the present disclosure provides a liposome composition comprising a PEGylated, non-targeted liposome encapsulating an ionizable carotenoid salt. In some embodiments, the liposome encapsulates the ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the PEGylated, non-targeted liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of ionizable carotenoid molecules. In another embodiment, the PEGylated, non-targeted liposome comprises a plurality of carotenoids. In a further embodiment, the liposome comprises a plurality of ionizable carotenoids, such as one or more ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D Two or more of the ionizable carotenoids provided in.

[0324] In other embodiments, the present disclosure provides liposome compositions comprising unpegylated liposomes encapsulating ionizable carotenoid salts. In some embodiments, the liposomes encapsulate the ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the liposomes contain less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, less than 10,000, or less than 5,000 ionizable carotenoid molecules. In some embodiments, the unpegylated liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range of ionizable carotenoid molecules therebetween. In other embodiments, the unpegylated liposomes contain a plurality of carotenoids. In a further embodiment, the liposomes comprise a plurality of ionizable carotenoids, such as those described herein [1]-

[28] and / or Figures 1A-1D Two or more of the ionizable carotenoids provided in.

[0325] In other embodiments, the present disclosure provides a liposome composition comprising a non-PEGylated and targeted liposome encapsulating an ionizable carotenoid salt, and encapsulating an ionizable carotenoid salt. In some embodiments, the liposome encapsulates an ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the non-PEGylated and targeted liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the non-PEGylated and targeted liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range of ionizable carotenoid molecules therebetween. In other embodiments, the non-PEGylated and targeted liposome contains a plurality of carotenoids. In a further embodiment, the liposomes comprise a plurality of ionizable carotenoids, such as those described herein [1]-

[28] and / or Figures 1A-1D Two or more of the ionizable carotenoids provided in.

[0326] In another embodiment, the present disclosure provides a liposome composition comprising an unpegylated and untargeted liposome encapsulating an ionizable carotenoid salt, and encapsulating an ionizable carotenoid salt. In some embodiments, the liposome encapsulates an ionizable carotenoid salt of any one of [1]-

[28] . In some embodiments, the unpegylated and untargeted liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of ionizable carotenoid molecules. In another embodiment, the unpegylated and untargeted liposome comprises a plurality of carotenoids. In a further embodiment, the liposome comprises a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D Two or more of the ionizable carotenoids provided in.

[0327] In some embodiments, the present disclosure provides a PEGylated liposome composition comprising liposomes encapsulating trans-crocetate. In some embodiments, the liposomes contain less than 6 million In some embodiments, the PEGylated liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of trans-crocetin molecules. In other embodiments, the liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more ionizable carotenoids provided in.

[0328] In other embodiments, the present disclosure provides liposome compositions comprising PEGylated and targeted liposomes encapsulating trans-crocetin. In some embodiments, the PEGylated and targeted liposomes contain 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range therebetween, targeting moieties. In some embodiments, the PEGylated and targeted liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-crocetin molecules. In other embodiments, the PEGylated and targeted liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more ionizable carotenoids provided in.

[0329] In other embodiments, the present disclosure provides liposome compositions comprising PEGylated and non-targeted liposomes encapsulating trans-crocetin. In some embodiments, the PEGylated and non-targeted liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-crocetin molecules. In other embodiments, the PEGylated and non-targeted liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more ionizable carotenoids provided in.

[0330] In other embodiments, the present disclosure provides liposome compositions comprising unpegylated liposomes encapsulating trans-crocetate. In some embodiments, the liposomes contain less than 6 million In some embodiments, the unPEGylated liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of trans-crocetin molecules. In other embodiments, the unPEGylated liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more ionizable carotenoids provided in.

[0331] In other embodiments, the present disclosure provides liposome compositions comprising unpegylated and targeted liposomes encapsulating trans-crocetin. In some embodiments, the unpegylated and targeted liposomes contain 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range therebetween, targeting moieties. In some embodiments, the unpegylated and targeted liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-crocetin molecules. In other embodiments, the unpegylated and targeted liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more carotenoids provided in.

[0332] In other embodiments, the present disclosure provides liposome compositions comprising unpegylated and non-targeted liposomes encapsulating trans-crocetin. In some embodiments, the unpegylated and non-targeted liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-crocetin molecules. In other embodiments, the unpegylated and non-targeted liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more carotenoids provided in.

[0333] In some embodiments, the present disclosure provides a PEGylated liposome composition comprising a liposome encapsulating a trans-norbixin salt. In some embodiments, the liposome contains less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, less than 10,000, or less than 5,000 trans-norbixin molecules. In some embodiments, the PEGylated liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of trans-norbixin molecules. In other embodiments, the liposome contains a plurality of carotenoids. In further embodiments, the liposome contains a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more ionizable carotenoids provided in.

[0334] In other embodiments, the present disclosure provides liposome compositions comprising PEGylated and targeted liposomes encapsulating trans-norbixin salts. In some embodiments, the PEGylated and targeted liposomes contain 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range therebetween, targeting moieties. In some embodiments, the PEGylated and targeted liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-norbixin molecules. In other embodiments, the PEGylated and targeted liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more ionizable carotenoids provided in.

[0335] In other embodiments, the present disclosure provides liposome compositions comprising PEGylated and non-targeted liposomes encapsulating trans-norbixin salts. In some embodiments, the PEGylated and non-targeted liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-norbixin molecules. In other embodiments, the PEGylated and non-targeted liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more ionizable carotenoids provided in.

[0336] In other embodiments, the present disclosure provides liposome compositions comprising unpegylated liposomes encapsulating trans-norbixin salt. In some embodiments, the liposomes contain less than 6 million In some embodiments, the unPEGylated liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, of trans-norbixin molecules. In other embodiments, the unPEGylated liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more ionizable carotenoids provided in.

[0337] In other embodiments, the present disclosure provides liposome compositions comprising unpegylated and targeted liposomes encapsulating trans-norbixin salts. In some embodiments, the unpegylated and targeted liposomes contain 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range therebetween, targeting moieties. In some embodiments, the unpegylated and targeted liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-norbixin molecules. In other embodiments, the unpegylated and targeted liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more carotenoids provided in.

[0338] In other embodiments, the present disclosure provides liposome compositions comprising unpegylated and non-targeted liposomes encapsulating trans-norbixin salts. In some embodiments, the unpegylated and non-targeted liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-norbixin molecules. In other embodiments, the unpegylated and non-targeted liposomes contain a plurality of carotenoids. In further embodiments, the liposomes contain a plurality of ionizable carotenoids, such as described herein [1]-

[28] and / or Figures 1A-1D One or more carotenoids provided in.

[0339] In some embodiments, provided liposome compositions include encapsulated one or more ionizable carotenoid salts (e.g., [1]-

[28] and / or Figures 1A-1D In some embodiments, the liposome composition comprises a divalent, trivalent or tetravalent salt of an ionizable carotenoid as described in any one of the preceding claims) and one or more aqueous pharmaceutically acceptable carriers. In some embodiments, the liposome composition contains trehalose. In some embodiments, the liposome composition contains 1% to 50% by weight of trehalose. In some embodiments, the liposome composition contains HBS at a concentration between 1 and 200 mM and a pH between 2 and 8. In some embodiments, the pH of the liposome composition is 5-8 or any range therebetween. In some embodiments, the pH of the liposome composition is 6-7 or any range therebetween.

[0340] In a further embodiment, the liposome composition provided comprises a liposome encapsulating trans-crocetin and one or more aqueous pharmaceutically acceptable carriers. In some embodiments, the liposome solution contains trehalose. In some embodiments, the liposome solution contains 1% to 50% weight trehalose. In some embodiments, the liposome solution contains HBS at a concentration of 1 to 200 mM, and a pH value of 2-8, or any range therebetween. In some embodiments, the pH value of the liposome solution is 5-8, or any range therebetween. In some embodiments, the pH value of the liposome solution is 6-7, or any range therebetween. In some embodiments, the trans-crocetin provided is a multivalent salt (e.g., divalent, trivalent or tetravalent). In some embodiments, the trans-crocetin is CTC. In some embodiments, the trans-crocetin is MTC.

[0341] In a further embodiment, the liposome composition provided comprises a liposome encapsulating a trans-norbixin salt and one or more aqueous pharmaceutically acceptable carriers. In some embodiments, the liposome solution contains trehalose. In some embodiments, the liposome solution contains 1% to 50% by weight of trehalose. In some embodiments, the liposome solution contains HBS at a concentration of 1 to 200 mM, and a pH value of 2-8, or any range therebetween. In some embodiments, the pH value of the liposome solution is 5-8, or any range therebetween. In some embodiments, the pH value of the liposome solution is 6-7, or any range therebetween. In some embodiments, the trans-norbixin salt provided is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the trans-norbixin salt is CTN. In some embodiments, the trans-norbixin salt is MTN.

[0342] The liposome provided comprises an aqueous compartment closed by at least one lipid bilayer. When the lipid comprising a hydrophilic head group is dispersed in water, they can spontaneously form a double-layer membrane called a lamella (lamellae). The lamella is composed of two single-layer lipid molecule sheets, and their non-polar (hydrophobic) surfaces are relative to each other, and their polar (hydrophilic) surfaces face the aqueous medium. The term "liposome" includes a single-layer vesicle, which is composed of a single lipid bilayer and generally has a diameter of about 20 to about 500nm, about 50 to about 300nm, about 50 to about 150nm, about 30 to about 1000nm, about 30 to about 175nm, about 80 to about 400nm, or about 80 to about 120nm. Liposome can also be multilayer, and it generally has an internal diameter within the range of 0.5 to 10um, wherein there are any number of concentric lipid bilayers from two to hundreds of alternating with aqueous layers. In some embodiments, the liposomes may include multilamellar vesicles (MLVs), large unilamellar vesicles (LUVs), and small unilamellar vesicles (SUVs). The lipids of the liposomes may be cationic, zwitterionic, neutral, or anionic, or any mixture thereof.

[0343] Any suitable lipid combination can be used to provide liposome and lipid nanoparticle provided herein.Lipid composition can be customized to affect characteristics, such as leakage rate, stability, granularity (for example, liposome diameter), zeta potential, protein binding, internal circulation and / or accumulation in tissue or organ.For example, DSPC and / or cholesterol can be utilized to reduce leakage from liposome.Negatively charged or positively charged lipids, such as DSPG and / or DOTAP can be included, to affect the surface charge of liposome or lipid nanoparticle.In some embodiments, lipid composition can include about ten kinds or less types of lipids, or about five kinds or less types of lipids, or about three kinds or less types of lipids.In some embodiments, the molar percentage (mol%) of the specific type of lipid present usually accounts for about 0% to about 10%, about 10% to about 30%, about 30% to about 50%, about 50% to about 70%, about 70% to about 90%, about 90% to about 100% of the total lipids present in liposome or lipid nanoparticle. In some embodiments, the therapeutic liposomes comprise 40-80 mol% DSPC, 5-50 mol% cholesterol, 0-30 mol% DSPG, and 0-10 mol% DSPE-PEG (2000). In some embodiments, the offensive liposomes comprise 40-70 mol% DPPC, 5-20 mol% cholesterol, 0-20 mol% DOTAP, and 20-40 mol% TPGS.

[0344] According to the required application, the particle size (diameter) of liposome can be adjusted.For example, when it is intended that liposome is delivered to cancerous tissue or inflamed tissue by permeability and retention enhancement (Enhanced Permeability and Retention, EPR) effect as injection product or similar product, preferably the liposome diameter is 20-500nm, 30-175nm or 50-150nm or any scope therebetween.In the case where liposome is intended to be delivered to macrophage, preferably the liposome diameter is 30 to 1000nm, or 80 to 400nm, or any scope therebetween.In the case where liposome composition is used as oral preparation or transdermal preparation, the particle size of liposome can be set to several microns.It should be noted that in normal tissue, vascular wall is used as barrier (because vascular wall is densely composed of vascular endothelial cells), and microparticles of specified size such as supramolecules and liposomes cannot be distributed in this tissue. However, in diseased tissues, the vessel walls are porous (because of the presence of interstitial spaces between endothelial cells), vascular permeability is increased, and supramolecules and microparticles can be distributed into extravascular tissues (increased permeability). Furthermore, the lymphatic system is well developed in normal tissues, but it is known that the lymphatic system is not developed in diseased tissues, and supramolecules or microparticles, once incorporated, are not recirculated through the systemic system and are retained in diseased tissues (enhanced retention), which forms the basis of the EPR effect (Wang et al., Ann. Rev. Med. 63: 185-198 (2012); Peer et al., Nat. Nanotech. 2: 751-760 (2007); Gubernator, Exp. Opin. Drug Deliv. 8: 565-580 (2011); Huwyler et al., Int. J. Nanomed. 3: 21-29 (2008); Maruyama et al. Adv. Drug Deliv. Rev. 63: 161-169 (2011); Musacchio and Torchilin ... Front. Biosci. 16: 1388-1412 (2011); Baryshnikov Vest. Ross. Akad. Med. Nauk. 23-31 (2012); and Torchilin Nat. Rev. Drug Disc. 4: 145-160 (2005). Therefore, it is possible to control the pharmacokinetics of liposomes by adjusting the particle size (diameter) of liposomes.

[0345] The size of the liposome in the provided liposome composition can vary between, for example, 0.5nm to 10um, or 20nm to 5um, depending on the phospholipid composition, the method used for their preparation and the expected therapeutic use of the liposome. In some embodiments, the median diameter of the liposome in the provided liposome composition is 20nm to 500nm, 50nm to 200nm, or 20nm to 200nm, or any range therebetween. In some embodiments, the median diameter of the liposome is 80nm to 120nm, or any range therebetween (e.g., 85-115nm, 90-110nm, 95-110nm or 95-105nm). In some embodiments, the median diameter of the liposomes in the provided liposome compositions is 10-250 nm, or any range therebetween (e.g., 10-225 nm, 10-200 nm, 10-175 nm, 10-150 nm, 40-150 nm, 50-150 nm, 60-150 nm, 70-150 nm, 80-150 nm, 90-150 nm, 100-150 nm, 10-1 In some embodiments, the median diameter of the liposomes in the liposome compositions provided is 100-250 nm, or any range therebetween (e.g., 100-225 nm, 100-200 nm, 100-175 nm, or 100-150 nm). In other embodiments, the median diameter of the liposomes in the liposome compositions provided is 10-100 nm, or any range therebetween (e.g., about 10-90 nm, 10-80 nm, 10-70 nm, 10-60 nm, or 10-50 nm). In some embodiments, the median diameter of the liposomes in the liposome compositions provided is less than about 500 nm, 450 nm, 400 nm, 350 nm, 300 nm, 250 nm, 200 nm, 150 nm, 145 nm, 150 nm, 135 nm, 130 nm, 125 nm, 120 nm, 115 nm, 110 nm, 105 nm, 100 nm, 95 nm, 90 nm, 85 nm, 80 nm, 75 nm, 70 nm, 65 nm, 60 nm, 55 nm, or 50 nm, 45 nm, or 40 nm. Dynamic laser scattering is a method well known to those skilled in the art for measuring liposome diameters.The diameter of liposomes (DLP) can be routinely determined using any technique and equipment known in the art, including, for example, dynamic laser light scattering (Coulter N4 particle size analyzer), Zetasizer Nano ZSP (Malvern, UK), and ELS-8000 (Otsuka Electronics Co., Ltd.)).

[0346] In some embodiments, the liposome compositions provided have a monodisperse particle size (diameter) distribution. "Monodisperse" and "uniform particle size distribution" are used interchangeably herein, and describe a number of liposome nanoparticles or microparticles wherein the particles have the same or nearly the same diameter. As used herein, a monodisperse distribution refers to a particle distribution wherein 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 86%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% or more of the particle distribution is located within 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15% or 10% of the mass median diameter.

[0347] In some embodiments, the liposome population in the liposome composition provided is relatively uniform. In some embodiments, the liposome population in the liposome composition provided is non-uniform. The polydispersity index can be used to represent the uniformity of the nanoparticle composition, for example, the particle size (diameter) distribution of the nanoparticle composition. A small (e.g., less than 0.3) polydispersity index generally represents a narrow particle size distribution. In some embodiments, the polydispersity index of the liposome population in the liposome composition provided is 0 to 0.25, or 0.01 to 0.1, or any range therebetween (e.g., 0.001 to 0.2, 0.005 to 0.1, 0.005 to 0, 0.005 to 0.09, 0.009 to 0.09, 0.01 to 0.08, 0.02 to 0.09, or 0.02 to 0.07, or any range therebetween).

[0348] In some embodiments, the liposomes in a population of liposomes in provided liposome compositions differ in their lipid composition, molar ratio of lipid components, size, charge (zeta potential), targeting ligand, and / or combinations thereof.

[0349] The zeta potential of a nanoparticle composition can be used to represent the zeta potential of the composition. For example, the zeta potential can describe the surface charge of a nanoparticle composition. It is generally desirable to have a nanoparticle composition with a relatively low positive or negative charge, because higher charged species may interact undesirably with cells, tissues, and other elements in the body. In some embodiments, the zeta potential of the nanoparticle composition can be about -10 mV to about +20 mV, about -10 mV to about +15 mV, about -10 mV to about +10 mV, about -10 mV to about +5 mV, about -10 mV to about 0 mV, about -10 mV to about -5 mV, about -5 mV to about +20 mV, about -5 mV to about +15 mV, about -5 mV to about +10 mV, about -5 mV to about +5 mV, about -5 mV to about 0 mV, about 0 mV to about +20 mV, about 0 mV to about +15 mV, about 0 mV to about +10 mV, about 0 mV to about +5 mV, about +5 mV to about +20 mV, about +5 mV to about +15 mV, or about +5 mV to about +10 mV. The zeta potential of liposomes can be routinely determined using techniques and equipment known in the art, including, for example, dynamic light scattering (Zetasizer Nano ZSP, Malvern, UK) and laser Doppler electrophoresis.

[0350] The encapsulation efficiency of therapeutic and / or prophylactic drugs such as ionizable carotenoids (e.g., trans-crocetin) describes the ratio of the amount of therapeutic and / or prophylactic drugs that are encapsulated or otherwise associated with the nanoparticle composition after preparation to the initial amount provided. It is desirable that the encapsulation efficiency is high (e.g., close to 100%). The encapsulation efficiency can be measured, for example, by comparing the amount of the therapeutic and / or prophylactic drugs in a solution containing the nanoparticle composition before and after removing the unencapsulated therapeutic and / or prophylactic drugs. For the liposome compositions described herein, the encapsulation efficiency of ionizable carotenoids (e.g., trans-crocetin) can be at least 50%, such as 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%. In some embodiments, the encapsulation efficiency is at least 80%. In certain embodiments, the encapsulation efficiency is at least 90%. In certain embodiments, the encapsulation efficiency is at least 95%. In certain embodiments, the encapsulation efficiency is at least 98%.

[0351] In other embodiments, the liposome composition provided contains the liposome of the salt of encapsulating ionizable carotenoid. In some embodiments, the ionizable carotenoid (for example, trans-crocetin) / lipid ratio of the liposome composition provided is 1 to 1000g / mol or any scope therebetween. In some embodiments, the ionizable carotenoid / lipid ratio of the liposome composition provided is 10 to 200g / mol, 10 to 150g / mol, 10 to 100g / mol, 20 to 200g / mol, 20 to 150g / mol, 20 to 100g / mol, 30 to 200g / mol, 30 to 150g / mol, 30 to 100g / mol, 40 to 200g / mol, 40 to 150g / mol, 40 to 100g / mol, 50 to 200g / mol, 50 to 150g / mol or 50 to 100g / mol, or any scope therebetween. In some embodiments, the ionizable carotenoid / lipid ratio is 30 to 90 g / mol, or any range therebetween. In some embodiments, the ionizable carotenoid / lipid ratio is 30 to 50 g / mol, 40 to 60 g / mol, 50 to 70 g / mol, 60 to 80 g / mol, or 70 to 90 g / mol, or any range therebetween. In some embodiments, the liposomes encapsulate the ionizable carotenoid salt of any one of [1] to

[27] . In some embodiments, the liposomes encapsulate Figures 1A-1D Any ionizable carotenoid salt listed in .

[0352] In some embodiments, the liposome composition contains liposomes encapsulating trans-crocetin. In some embodiments, the trans-crocetin / lipid ratio of the liposome composition provided is 1 to 1000 g / mol, or any range therebetween. In some embodiments, the trans-crocetin / lipid ratio of the liposome composition is 10 to 200 g / mol, 10 to 150 g / mol, 10 to 100 g / mol, 20 to 200 g / mol, 20 to 150 g / mol, 20 to 100 g / mol, 30 to 200 g / mol, 30 to 150 g / mol, 30 to 100 g / mol, 40 to 200 g / mol, 40 to 150 g / mol, 40 to 100 g / mol, 50 to 200 g / mol, 50 to 150 g / mol, or 50 to 100 g / mol, or any range therebetween. In some embodiments, the trans-crocetin / lipid ratio is 30 to 90 g / mol, or any range therebetween. In some embodiments, the trans-crocetin / lipid ratio is 30 to 50 g / mol, 40 to 60 g / mol, 50 to 70 g / mol, 60 to 80 g / mol, or 70 to 90 g / mol, or any range therebetween.

[0353] In some embodiments, the liposome composition contains liposomes encapsulating trans-norbixin salts. In some embodiments, the trans-norbixin / lipid ratio of the liposome composition provided is 1 to 1000 g / mol, or any range therebetween. In some embodiments, the trans-norbixin / lipid ratio of the liposome composition provided is 10 to 200 g / mol, 10 to 150 g / mol, 10 to 100 g / mol, 20 to 200 g / mol, 20 to 150 g / mol, 20 to 100 g / mol, 30 to 200 g / mol, 30 to 150 g / mol, 30 to 100 g / mol, 40 to 200 g / mol, 40 to 150 g / mol, 40 to 100 g / mol, 50 to 200 g / mol, 50 to 150 g / mol, or 50 to 100 g / mol, or any range therebetween. In some embodiments, the trans-norbixin / lipid ratio is 30 to 90 g / mol, or any range therebetween. In some embodiments, the trans-norbixin / lipid ratio is 30 to 50 g / mol, 40 to 60 g / mol, 50 to 70 g / mol, 60 to 80 g / mol, or 70 to 90 g / mol, or any range therebetween.

[0354] In some embodiments, the liposome composition is buffered using a zwitterionic buffer. Suitably, the zwitterionic buffer is an aminoalkanesulfonic acid or a suitable salt. Examples of aminoalkanesulfonic acid buffers include, but are not limited to, HEPES, HEPPS / EPPS, MOPS, MOBS, and PIPES. Preferably, the buffer is a pharmaceutically acceptable buffer suitable for use in humans, such as for commercial injectable products. Most preferably, the buffer is HEPES. The liposome composition may suitably include AGP.

[0355] In some embodiments, the liposome composition is buffered with HEPES. In some embodiments, the liposome composition is buffered with HEPES having a pH of about 7.

[0356] In some embodiments, the pharmaceutical composition is a liposome composition comprising cationic liposomes. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero. In some embodiments, the liposome has a zeta potential of 0.2 to 150mV, 1 to 50mV, 1 to 40mV, 1 to 30mV, 1 to 25mV, 1 to 20mV, 1 to 15mV, 1 to 10mV, 1 to 5mV, 2 to 10mV, 3 to 10mV, 4 to 10mV, or 5 to 10mV, or any range therebetween. In some embodiments, the liposome has a diameter of 20nm to 500nm, 20nm to 200nm, 30nm to 175nm, 50nm to 200nm, or 50nm to 150nm, or any range therebetween. In some embodiments, the cationic liposome has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the liposome composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75% or greater than 75% w / w of an ionizable carotenoid (e.g., trans-crocetin). In some embodiments, during the preparation of the liposome composition, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 75%, 80%, 85%, 90%, 95% or 97% of the ionizable carotenoid starting material is encapsulated (embedded) in the liposomes of the liposome composition. In additional embodiments, the ionizable carotenoid (eg, trans-crocetin) encapsulated by the liposomes is in a HEPES buffered solution within the liposomes. In further embodiments, the liposomes comprise at least one OxPAPC.

[0357] In some embodiments, the pharmaceutical composition provided is a liposome composition comprising anionic or neutral liposomes. In some embodiments, the liposome composition comprises a liposome having a zeta potential less than or equal to zero. In some embodiments, the liposome has a zeta potential of -150 to 0, -50 to 0mV, -40 to 0mV, -30 to 0mV, -25 to 0mV, -20 to 0mV, -10 to 0mV, -9 to 0mV, -8 to 0mV, -7 to 0mV, -6 to 0mV, -5 to 0mV, -4 to 0mV, -3 to 0mV, -2 to 0mV, -1 to 0mV, or -8 to 2mV, or any range therebetween. In some embodiments, the anionic or neutral liposome has a diameter of 20nm to 500nm, 20nm to 200nm, 30nm to 175nm, or 50nm to 150nm, or any range therebetween. In other embodiments, the anionic or neutral liposome has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the anionic liposome has a diameter of 20nm to 500nm, 20nm to 200nm, 30nm to 175nm, or 50nm to 150nm, or any range therebetween. In further embodiments, the anionic liposome has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the neutral liposome has a diameter of 20nm to 500nm, 20nm to 200nm, 30nm to 175nm, or 50nm to 150nm, or any range therebetween. In some embodiments, the neutral liposome has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the pharmaceutical composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or greater than 75% w / w of an ionizable carotenoid (e.g., trans-crocetin). In some embodiments, during the preparation of the liposome composition, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or greater than 75% of the starting material of the ionizable carotenoid is encapsulated (embedded) in the liposome. In some embodiments, the liposome composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or greater than 75% w / w ionizable carotenoid (e.g., trans-crocetin).In some embodiments, the anionic or neutral liposome composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more than 75% w / w of ionizable carotenoids (e.g., trans-crocetin). In some embodiments, the liposome composition comprises at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more than 75% w / w of ionizable carotenoids (e.g., trans-crocetin). In other embodiments, the ionizable carotenoids (e.g., trans-crocetin) encapsulated by anionic or neutral liposomes are in a HEPES buffer solution within the liposome. In a further embodiment, the liposome comprises at least one OxPAPC.

[0358] In some embodiments, the pharmaceutical composition provided is a liposome composition comprising liposomes containing at least one OxPAPC. In some embodiments, the OxPAPC is a phospholipid that is oxidized and / or contains fragmented oxygenated sn-2 residues. In some embodiments, the OxPAPC is an oxidized phospholipid containing a five-carbon sn-2 residue with an ω-aldehyde or ω-carboxyl group. In some embodiments, the OxPAPC is an oxidized phospholipid selected from HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC. In some embodiments, the OxPAPC is a phospholipid containing epoxyisoprostane. In some embodiments, the OxPAPC is PGPC. In some embodiments, the liposomes contain at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30% OxPAPC. In some embodiments, the liposome composition has cationic liposomes comprising 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, or any range therebetween of OxPAPC. In some embodiments, the liposome composition comprises cationic liposomes. In some embodiments, the liposome composition comprises neutral liposomes. In some embodiments, the liposome composition comprises anionic liposomes. In other embodiments, the liposome composition comprises at least one liposome containing OxPAPC having a diameter of 20nm to 500nm, 20nm to 200nm, 30nm to 175nm, or 50nm to 150nm, or any range therebetween. In further embodiments, the liposome composition comprises at least one liposome containing OxPAPC having a diameter of 80nm to 120nm, or any range therebetween.

[0359] In some embodiments, the pharmaceutical compositions provided are liposome compositions comprising cationic liposomes comprising at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30% OxPAPC. In some embodiments, the liposome composition has cationic liposomes comprising 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, or any range therebetween of OxPAPC. In some embodiments, the liposomes comprise at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30% OxPAPC. In some embodiments, the liposome composition has cationic liposomes containing about 10% OxPAPC. In some embodiments, the liposome composition has cationic liposomes comprising at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30% PGPC. In some embodiments, the liposomes comprise 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, or any range therebetween of PGPC. In some embodiments, the liposome composition has cationic liposomes comprising about 10% PGPC.

[0360] In some embodiments, the pharmaceutical composition is a liposome composition comprising anionic or neutral liposomes comprising at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30% OxPAPC. In some embodiments, the liposome composition has anionic or neutral liposomes comprising 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, or any range therebetween of OxPAPC. In some embodiments, the liposomes comprise at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30% OxPAPC. In some embodiments, the liposome composition has anionic or neutral liposomes containing about 10% OxPAPC. In some embodiments, the liposome composition has anionic or neutral liposomes comprising at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30% PGPC. In some embodiments, the liposomes comprise 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, or any range of PGPC therebetween. In some embodiments, the liposome composition has anionic or neutral liposomes containing about 10% PGPC.

[0361] In some embodiments, the pharmaceutical composition provided is a liposome composition comprising neutral liposomes, wherein the neutral liposomes contain at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30% OxPAPC. In some embodiments, the liposome composition containing neutral OxPAPC contains 0.01%-35%, 0.1%-30%, 1%-25%, 3-20% or 5-15% OxPAPC or any range therebetween. In some embodiments, the liposome composition containing neutral OxPAPC contains about 10% OxPAPC. In some embodiments, the liposome composition containing neutral OxPAPC contains at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30% PGPC. In some embodiments, the liposome composition containing neutral PGPC comprises 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15% PGPC or any range therebetween. In some embodiments, the liposome composition containing neutral OxPAPC comprises about 10% PGPC.

[0362] In additional embodiments, the liposomes in the liposome composition are pegylated.

[0363] In some embodiments, the pharmaceutical composition provided is a non-targeted liposome composition. That is, the liposomes in the liposome composition have no specific affinity for an epitope expressed on the surface of a target cell of interest (e.g., an epitope on a surface antigen). In a further embodiment, the non-targeted liposome composition is pegylated.

[0364] In some cases, the accumulation of liposomes at the target site may be due to the enhancement of the permeability and retention characteristics of certain tissues (such as cancerous tissues). Accumulation in this manner is often partly due to the size of the liposomes, and specific targeting functionality may not be required. In other embodiments, the liposomes provided include a targeting agent. Generally speaking, the targeting agent can be associated with any target of interest, such as a target associated with an organ, tissue, cell, extracellular matrix or intracellular region. In certain embodiments, the target may be associated with a specific disease state such as a cancerous condition. In some embodiments, the targeting component can be specific only to a target such as a receptor. Suitable targets may include but are not limited to nucleic acids, such as DNA, RNA or its modified derivatives. Suitable targets may also include but are not limited to proteins, such as extracellular proteins, receptors, cell surface receptors, tumor markers, transmembrane proteins, enzymes or antibodies. Suitable targets may include carbohydrates, such as monosaccharides, disaccharides or polysaccharides that may be present on the cell surface, for example.

[0365] In certain embodiments, the targeting agent may include a target ligand (e.g., a peptide containing RGD), a small molecule mimetic of a target ligand (e.g., a peptide mimetic ligand), or an antibody or antibody fragment specific to a particular target. In some embodiments, the targeting agent may further include a folic acid derivative, a B-12 derivative, an integrin RGD peptide, an NGR derivative, a somatostatin derivative or peptide that binds to a somatostatin receptor, for example, octreotide and octreotate, etc. In some embodiments, the targeting agent includes an aptamer. An aptamer may be designed to associate or bind to a target of interest. An aptamer may be composed of, for example, DNA, RNA, and / or a peptide, and certain aspects of an aptamer are known in the art. (See, for example, Klussman, The Aptamer Handbook, Wiley-VCH (2006); Nissenbaum, Trends in Biotech. 26 (8): 442-449 (2008)).

[0366] In other embodiments, the liposome composition comprises a targeted liposome. That is, the liposome contains a targeting moiety with specific affinity for an epitope (e.g., a surface antigen or other molecule) on a target cell of interest. In some embodiments, the targeting moiety of the liposome is not attached to the liposome by a covalent bond. In other embodiments, the targeting moiety of the liposome is attached to one or both of PEG and the outside of the liposome. In a further embodiment, the targeted liposome is pegylated. The function of the targeting moiety of the targeted liposome may include, but is not limited to, targeting the liposome to a target cell of interest in vivo or in vitro; interacting with a surface antigen to which the targeting moiety has a specific affinity, and delivering the liposome payload (e.g., trans-crocetin) to the location of the cell or into the cell.

[0367] Suitable targeting moieties are known in the art and include, but are not limited to, antibodies, antigen-binding antibody fragments, scaffold proteins, polypeptides, and peptides. In some embodiments, the targeting moiety is a polypeptide. In further embodiments, the targeting moiety is a polypeptide comprising at least 3, 5, 10, 15, 20, 30, 40, 50, or 100 amino acid residues. In some embodiments, the targeting moiety is an antibody or an antigen-binding antibody fragment. In further embodiments, the targeting moiety comprises one or more of an antibody, a humanized antibody, an antigen-binding fragment of an antibody, a single-chain antibody, a single-domain antibody, a bispecific antibody, a synthetic antibody, a pegylated antibody, and a multimeric antibody. In some embodiments, the targeting moiety has a specific affinity for an epitope that is preferentially expressed on a target cell, such as a tumor cell, compared to a normal or non-tumor cell. In some embodiments, the targeting moiety has a specific affinity for an epitope on a tumor cell surface antigen that is present on a tumor cell but is absent or inaccessible on a non-tumor cell. In some embodiments, the targeting moiety binds to an epitope of interest with an equilibrium dissociation constant (Kd) such as that obtained using The analysis was performed at 50 x 10 -12 Up to 10x 10 -6 In a further embodiment, the Kd is determined using a surface plasmon resonance technique in which an antigen containing the epitope is immobilized, a targeting moiety is used as the analyte, and the following conditions are used: 10 mM MES buffer, 0.05% polyoxyethylene sorbitan monolaurate, and 150 mM NaCl at 37°C.

[0368] In certain embodiments, the targeting moiety comprises a polypeptide that specifically binds to a folate receptor. In some embodiments, the targeting moiety is an antibody or an antigen-binding antibody fragment. In some embodiments, the targeting moiety is a folate (e.g., a folate-PEG conjugate) or a folate derivative conjugated to the surface of a liposome. In some embodiments, the folate receptor bound by the targeting moiety is one or more folate receptors selected from the following: folate receptor α (FR-α, FOLR1), folate receptor β (FR-β, FOLR2) and folate receptor δ (FR-δ, FOLR4). In some embodiments, the folate receptor bound by the targeting moiety is folate receptor α (FR-α). In some embodiments, the folate receptor bound by the targeting moiety is folate receptor β (FR-β). In some embodiments, the targeting moiety specifically binds to FR-α and FR-β.

[0369] In other embodiments, the liposome composition comprises one or more of an immunostimulant, a detectable marker, and a maleimide, which is arranged on at least one of the PEG and the outside of the liposome. In some embodiments, the liposome of the liposome composition is cationic. In other embodiments, the liposome of the liposome composition is anionic or neutral. In other embodiments, the liposome of the liposome composition has a diameter of 20nm to 500nm or any range therebetween. In further embodiments, the liposome of the liposome composition has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the liposome of the liposome composition is pegylated. In some embodiments, the liposome of the liposome composition is targeted. In further embodiments, the liposome of the liposome composition is pegylated and targeted.

[0370] In some embodiments, the present disclosure provides a pharmaceutical composition comprising a liposome-encapsulated ionizable carotenoid having the formula:

[0371] Polyene carotenoids-Q, wherein

[0372] The polyene carotenoids include

[0373] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds,

[0374] (b) a methyl or lower alkyl (C2-C3) substituent, and

[0375] (c) 1, 2, 3 or more ionizable groups; and

[0376] Q is (a) a multivalent counterion or (b) a monovalent cation.

[0377] In some embodiments, the polyene carotenoid comprises an all-trans conjugated double bond. In some embodiments, the polyene carotenoid comprises 6-9 conjugated double bonds. In a specific embodiment, the polyene carotenoid comprises 7 conjugated double bonds. The polyene carotenoid may be naturally occurring or synthetic. In some embodiments, the polyene carotenoid is naturally occurring. In other embodiments, the polyene carotenoid is synthetic. The one or more ionizable groups may be anionic and / or cationic. In some embodiments, the polyene carotenoid-Q comprises two or more identical ionizable groups. In some embodiments, the polyene carotenoid comprises an all-trans conjugated double bond. In some embodiments, the polyene carotenoid comprises 6-9 conjugated double bonds. In some embodiments, the polyene carotenoid comprises two or more different ionizable groups. In some embodiments, the polyene carotenoid-Q comprises one or more anionic ionizable groups. In some embodiments, the polyene carotenoid comprises at least one ionizable group selected from the group consisting of carboxyl, sulfonate, sulfate, phosphonate or phosphate and hydroxamate moieties. In other embodiments, the polyene carotenoid-Q comprises one or more cationic ionizable groups (e.g., primary, secondary or tertiary amine groups, quaternary ammonium groups, choline groups, guanidine groups or imidazolyl groups). In specific embodiments, the polyene carotenoid comprises at least one cationic ionizable group, and the pharmaceutical composition is substantially free of nucleic acids.

[0378] In some embodiments, Q is a multivalent counterion. In some embodiments, Q is a multivalent cationic counterion. In some embodiments, Q is a multivalent metal cation. In some embodiments, Q is a multivalent transition metal cation. In some embodiments, Q is a divalent counterion. In some embodiments, Q is a divalent cationic counterion. In some embodiments, Q is a divalent metal cation. In some embodiments, Q is a divalent transition metal cation. In some embodiments, Q is selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ In some embodiments, Q is Ca 2+ or Mg 2+ In some embodiments, Q is Ca 2+ In further embodiments, the polyene carotenoid-Q is trans-calcium crocetin (CTC). In some embodiments, Q is Mg 2+In further embodiments, the polyene carotenoid-Q is magnesium trans-crocetin (MTC). In other embodiments, Q is a trivalent cationic counterion, such as Fe 3+ In some embodiments, Q is a multivalent organic counterion. In some embodiments, Q is a divalent organic cation. In some embodiments, Q is a divalent organic cation, such as a protonated diamine.

[0379] In further embodiments, Q is a monovalent counterion. In some embodiments, Q is a monovalent cationic counterion. In some embodiments, Q is a monovalent metal cation. In some embodiments, Q is selected from Na + , Li + or K + In some embodiments, Q is an organic cation. In some embodiments, Q is a divalent organic cation. In some embodiments, Q is a monovalent organic cation, such as a protonated amine (e.g., a protonated diamine or a protonated polyamine). In some embodiments, Q is an organic cation, such as NH4 + , protonated diamines or protonated polyamines.

[0380] In some embodiments, the liposome contains less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, less than 10,000, or less than 5,000 ionizable carotenoid molecules. In some embodiments, the liposome contains ionizable carotenoid molecules between 10 and 100,000 or any range therebetween. In some embodiments, the ionizable carotenoid / lipid ratio of the liposome composition is 1 g / mol and about 1000 g / mol, or any range therebetween. In some embodiments, the ionizable carotenoid / lipid ratio is 10-150 g / mol, 10-100 g / mol, 30-200 g / mol, 40-200 g / mol, or 50-200 g / mol, or any range therebetween. In some embodiments, the liposome contains at least 0.1% to 97% trans-crocetin. In some embodiments, the liposome has a diameter of 20nm to 500nm, 20nm to 200nm, or 80nm to 120nm, or any range therebetween. In some embodiments, the liposome is formed by a liposome component. In a further embodiment, the liposome component comprises at least one of anionic lipids and neutral lipids. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol PEG, and cholesterol-maleimide. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, cholesterol, and HSPC. In another embodiment, the liposome further comprises an oxidized phospholipid, such as OxPAPC. In some embodiments, the liposome comprises OxPAPC, which is an oxidized phospholipid containing a fragmented oxygenated sn-2 residue, an oxidized phospholipid containing a full-length oxygenated sn-2 residue, and / or an oxidized phospholipid containing a five-carbon sn-2 residue with an ω-aldehyde or ω-carboxyl group. In some embodiments, the liposome comprises OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC, and KOOA-PC, or OxPAPC is a phospholipid containing epoxyisoprostane.In some embodiments, the liposome comprises an OxPAPC selected from the group consisting of 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxycyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxyisoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine ( In some embodiments, the liposome comprises PGPC; 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, OxPAPC within the lipid bilayer of the liposome is 0%-100% of the total lipids or any range therebetween. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen or other molecule on a target cell of interest. In some embodiments, the targeting moiety is attached to one or both of PEG and the outside of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the outside of the liposome by a covalent bond. In some embodiments, the targeting moiety is a polypeptide. In a further embodiment, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the liposome further comprises an immunostimulant (such as 1,6-β-glucan). In some embodiments, the liposome comprises a steric stabilizer. In some embodiments, the steric stabilizer is polyethylene glycol (ie, the liposome is pegylated). In some embodiments, the PEG has a number average molecular weight (Mn) of 200 to 5000 daltons. In other embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential less than or equal to zero. In some embodiments, the liposomes have a zeta potential of -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween.In other embodiments, the liposome is cationic. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero. In some embodiments, the liposome has a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween.

[0381] In some embodiments, the present disclosure provides a pharmaceutical composition comprising a liposome-encapsulated ionizable carotenoid having the formula:

[0382] Q-R1-polyene carotenoid-R2-Q, wherein

[0383] The polyene carotenoids include

[0384] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds,

[0385] (b) a methyl or lower alkyl (C2-C3) substituent, and

[0386] (c) 1, 2, 3 or more ionizable groups;

[0387] R1 and R2 are ionizable groups; and

[0388] Q is (a) a multivalent counterion or (b) a monovalent cation.

[0389] In some embodiments, the polyene carotenoid comprises an all-trans conjugated double bond. In a specific embodiment, the polyene carotenoid comprises 6-9 conjugated double bonds. The polyene carotenoid may be naturally occurring or synthetic. In some embodiments, the polyene carotenoid is naturally occurring. In other embodiments, the polyene carotenoid is synthetic. In some embodiments, R1 and R2 are the same ionizable groups. In other embodiments, R1 and R2 are different ionizable groups. In some embodiments, R1 and R2 are the same cationic ionizable groups. In other embodiments, R1 and R2 are different cationic groups. In some embodiments, R1 and R2 are the same anionic ionizable groups. In other embodiments, R1 and R2 are different anionic groups. In some embodiments, R1 is a cationic ionizable group or an anionic ionizable group, and R2 is an anionic ionizable group or a cationic group, respectively. In some embodiments, the polyene carotenoid comprises at least one anionic ionizable group. In some embodiments, the polyene carotenoid comprises at least one ionizable group selected from the following: carboxyl, sulfonate, sulfate, phosphonate or phosphate and hydroxamate moieties. In some embodiments, R1 is at least one ionizable group selected from the following: carboxyl, sulfonate, sulfate, phosphonate or phosphate and hydroxamate moieties. In some embodiments, R2 is at least one ionizable group selected from the following: carboxyl, sulfonate, sulfate, phosphonate or phosphate and hydroxamate moieties. In other embodiments, the polyene carotenoid-Q comprises one or more cationic ionizable groups (e.g., primary, secondary or tertiary amine groups, quaternary ammonium groups, choline groups, guanidine groups or imidazolyl groups). In specific embodiments, the polyene carotenoid comprises at least one cationic ionizable group, and the pharmaceutical composition is substantially free of nucleic acids.

[0390] In some embodiments, Q is a multivalent counterion. In some embodiments, Q is a multivalent cationic counterion. In further embodiments, Q is a multivalent metal cation. In some embodiments, Q is a multivalent transition metal counterion. In some embodiments, Q is a divalent counterion. In some embodiments, Q is a divalent cationic counterion. In further embodiments, Q is a divalent metal cation. In some embodiments, Q is selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2 + and Fe 2+ In a further embodiment, Q is Ca 2+ or Mg 2+In a further embodiment, Q is Ca 2+ In some embodiments, Q is Mg 2+ In other embodiments, Q is a trivalent cationic counterion such as Fe 3+ In other embodiments, Q is a multivalent organic counterion. In some embodiments, Q is a divalent organic cation. In some embodiments, Q is a divalent organic cation, such as a protonated diamine.

[0391] In some embodiments, Q is a monovalent cationic counterion. In further embodiments, Q is selected from Na + or Li + or K + In some embodiments, Q is an organic counterion. In some embodiments, Q is a multivalent organic cation. In further embodiments, Q is a divalent organic cation, such as a protonated diamine or a protonated polyamine. In other embodiments, Q is a monovalent organic cation, such as NH4 + , protonated diamines or protonated polyamines.

[0392] In some embodiments, the liposome contains less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, less than 10,000, or less than 5,000 ionizable carotenoid molecules. In some embodiments, the liposome contains ionizable carotenoid molecules between 10 and 100,000 or any range therebetween. In some embodiments, the ionizable carotenoid / lipid ratio of the liposome composition is 1 g / mol and about 1000 g / mol or any range therebetween. In some embodiments, the ionizable carotenoid / lipid ratio is 10-150 g / mol, 10-100 g / mol, 30-200 g / mol, 40-200 g / mol, or 50-200 g / mol, or any range therebetween. In some embodiments, the liposome contains at least 0.1% to 97% ionizable carotenoids. In some embodiments, the liposome has a diameter of 20nm to 500nm, 20nm to 200nm, or 80nm to 120nm, or any range therebetween. In some embodiments, the liposome is formed by a liposome component. In a further embodiment, the liposome component comprises at least one of anionic lipids and neutral lipids. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol PEG, and cholesterol-maleimide. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, cholesterol, and HSPC. In another embodiment, the liposome further comprises an oxidized phospholipid, such as OxPAPC. In some embodiments, the liposome comprises OxPAPC, which is an oxidized phospholipid containing a fragmented oxygenated sn-2 residue, an oxidized phospholipid containing a full-length oxygenated sn-2 residue, and / or an oxidized phospholipid containing a five-carbon sn-2 residue with an ω-aldehyde or ω-carboxyl group. In some embodiments, the liposome comprises OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC, and KOOA-PC, or OxPAPC is a phospholipid containing epoxyisoprostane.In some embodiments, the liposome comprises an OxPAPC selected from the group consisting of 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxycyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxyisoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine ( In some embodiments, the liposome comprises PGPC; 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, OxPAPC within the lipid bilayer of the liposome is 0%-100% of the total lipids or any range therebetween. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest. In some embodiments, the targeting moiety is attached to one or both of PEG and the outside of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the outside of the liposome by a covalent bond. In some embodiments, the targeting moiety is a polypeptide. In a further embodiment, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the liposome further comprises an immunostimulant (such as 1,6-β-glucan). In some embodiments, the liposome comprises a steric stabilizer. In some embodiments, the steric stabilizer is polyethylene glycol (ie, the liposome is pegylated). In some embodiments, the PEG has a number average molecular weight (Mn) of 200 to 5000 daltons. In some embodiments, the zeta potential of the liposome is -150 to 150mV, or -50 to 50mV, or in any range therebetween. In some embodiments, the liposomes are cationic. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero.In some embodiments, the liposomes have a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween. In some embodiments, the liposomes are anionic or neutral. In some embodiments, the liposome composition comprises liposomes having a zeta potential less than or equal to zero. In some embodiments, the liposomes have a zeta potential of -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween.

[0393] In some embodiments, the present disclosure provides a pharmaceutical composition comprising an ionizable di-α,ω-carotenoid having the formula:

[0394] Q-R1-polyene carotenoid-R1-Q, wherein the polyene carotenoid comprises

[0395] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds,

[0396] (b) a methyl or lower alkyl (C2-C3) substituent, and

[0397] (c) 1, 2, 3 or more ionizable groups; and

[0398] R1 is an ionizable group; and

[0399] Q is (a) a multivalent counterion or (b) a monovalent cation.

[0400] In some embodiments, R1-polyene carotenoid-R1 comprises all-trans conjugated double bonds. In some embodiments, the R1-polyene carotenoid-R1 comprises 6-9 conjugated double bonds. In a specific embodiment, R1-polyene carotenoid-R1 comprises 7 conjugated double bonds. R1-polyene carotenoid-R1 can be naturally occurring or synthetic. In some embodiments, R1-polyene carotenoid-R1 is naturally occurring. In other embodiments, the b R1-polyene carotenoid-R1 is synthetic. In some embodiments, R1 is an anionic ionizable group. In some embodiments, R1-polyene carotenoid-R1 comprises an ionizable group selected from the following: carboxyl, sulfonate, sulfate, phosphonate, phosphate and hydroxamate moieties. In other embodiments, R1 is a cationic ionizable group (e.g., primary, secondary or tertiary amine, quaternary ammonium, choline, guanidine or imidazole). In certain embodiments, R1 is a cationic ionizable group, and the pharmaceutical composition is substantially free of nucleic acids.

[0401] In some embodiments, Q is a multivalent counterion. In some embodiments, Q is a multivalent cationic counterion. In further embodiments, Q is a multivalent metal cation. In some embodiments, Q is a multivalent transition metal counterion. In some embodiments, Q is a divalent counterion. In some embodiments, Q is a divalent cationic counterion. In further embodiments, Q is a divalent metal cation. In some embodiments, Q is selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2 + and Fe 2+ In a further embodiment, Q is Ca 2+ or Mg 2+ In a further embodiment, Q is Ca 2+ In some embodiments, Q is Mg 2+ In other embodiments, Q is a trivalent cationic counterion such as Fe 3+ In other embodiments, Q is a multivalent organic counterion. In some embodiments, Q is a divalent organic cation. In some embodiments, Q is a divalent organic cation, such as a protonated diamine.

[0402] In some embodiments, Q is a monovalent cationic counterion. In further embodiments, Q is selected from Na + or Li + or K +In some embodiments, Q is an organic counterion. In some embodiments, Q is a multivalent organic cation. In further embodiments, Q is a divalent organic cation, such as a protonated diamine or a protonated polyamine. In other embodiments, Q is a monovalent organic cation, such as NH4 + , protonated diamines or protonated polyamines.

[0403] In some embodiments, the liposome contains less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000 or less than 10,000 ionizable carotenoid molecules. In some embodiments, the liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range of ionizable carotenoid molecules therebetween. In some embodiments, the ionizable carotenoid / lipid ratio of the liposome composition is 1 g / mol and about 1000 g / mol, or any range therebetween. In some embodiments, the ionizable carotenoid / lipid ratio is 10-150 g / mol, 10-100 g / mol, 30-200 g / mol, 40-200 g / mol, or 50-200 g / mol, or any range therebetween. In some embodiments, the liposome comprises at least 0.1% to 97% ionizable carotenoids. In some embodiments, the liposome has an average diameter of, for example, 20nm to 500nm (nanometers), or 20nm to 200nm, or any range therebetween. In some embodiments, the liposome has an average diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the liposome is formed by a liposome component. In a further embodiment, the liposome component comprises at least one of anionic lipids and neutral lipids. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol PEG, and cholesterol-maleimide. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, cholesterol, and HSPC. In other embodiments, the liposome further comprises an oxidized phospholipid, such as OxPAPC. In some embodiments, the liposome comprises OxPAPC, which is an oxidized phospholipid containing fragmented oxygenated sn-2 residues, an oxidized phospholipid containing full-length oxygenated sn-2 residues, and / or an oxidized phospholipid containing five-carbon sn-2 residues with ω-aldehyde or ω-carboxyl groups. In some embodiments, the liposome comprises OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC, or OxPAPC is a phospholipid containing epoxyisoprostanoids.In some embodiments, the liposome comprises an OxPAPC selected from the group consisting of 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxycyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxyisoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine ( In some embodiments, the liposome comprises PGPC; 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, OxPAPC within the lipid bilayer of the liposome is 0%-100% of the total lipids or any range therebetween. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest. In some embodiments, the targeting moiety is attached to one or both of PEG and the outside of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the outside of the liposome by a covalent bond. In some embodiments, the targeting moiety is a polypeptide. In a further embodiment, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the liposome further comprises an immunostimulant (such as 1,6-β-glucan). In some embodiments, the liposome comprises a steric stabilizer. In some embodiments, the steric stabilizer is polyethylene glycol (ie, the liposome is pegylated). In some embodiments, the PEG has a number average molecular weight (Mn) of 200 to 5000 daltons. In other embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential less than or equal to zero. In some embodiments, the liposomes have a zeta potential of -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween.In other embodiments, the liposome is cationic. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero. In some embodiments, the liposome has a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween.

[0404] In some embodiments, the present disclosure provides a pharmaceutical composition comprising a liposome-encapsulated bi-α,ω-carotenoid having the formula: R1-polyene carotenoid-R1, wherein,

[0405] The di-α,ω-carotenoids contain:

[0406] (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds, and

[0407] (b) 1, 2, 3 or more ionizable groups; and

[0408] The polyene carotenoid is optionally substituted with 1 to n methyl or lower C1-C3 alkyl substituents, wherein n=1 to 4; and

[0409] R1 is a polar group and / or a monocyclic functional group.

[0410] In some embodiments, the bis-α,ω-carotenoid comprises an all-trans conjugated double bond. In some embodiments, the bis-α,ω-carotenoid comprises 6-9 conjugated double bonds. In a specific embodiment, the bis-α,ω-carotenoid comprises 7 conjugated double bonds. The bis-α,ω-carotenoid may be naturally occurring or synthetic. In some embodiments, the bis-α,ω-carotenoid is naturally occurring. In other embodiments, the bis-α,ω-carotenoid is synthetic. In some embodiments, R1 is a polar group. In some embodiments, R1 is a monocyclic functional group. In some embodiments, R1 is a polar group and a monocyclic functional group. In some embodiments, the bis-α,ω-carotenoid comprises a monocyclic and / or polar functional group selected from the functional groups present in astaxanthin, lutein, lutein and zeaxanthin. In some embodiments, the bi-α,ω-carotenoid is selected from astaxanthin, lutein, lutein, and zeaxanthin (eg, as shown below).

[0411]

[0412] In some embodiments, the liposome contains less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000 or less than 10,000 bi-alpha, omega-carotenoid molecules. In some embodiments, the liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween of bi-alpha, omega-carotenoid molecules. In some embodiments, the bi-alpha, omega-carotenoid / lipid ratio of the liposome composition is 1 g / mol and about 1000 g / mol or any range therebetween. In some embodiments, the bi-alpha, omega-carotenoid / lipid ratio is 10-150 g / mol, 10-100 g / mol, 30-200 g / mol, 40-200 g / mol, or 50-200 g / mol, or any range therebetween. In some embodiments, the liposome comprises at least 0.1% to 97% of bi-α, ω-carotenoids. In some embodiments, the liposome has a diameter of 20nm to 500nm, or 20nm to 200nm, or any range therebetween. In some embodiments, the liposome has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the liposome is formed by a liposome component. In a further embodiment, the liposome component comprises at least one of anionic lipids and neutral lipids. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol PEG, and cholesterol-maleimide. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, cholesterol, and HSPC. In other embodiments, the liposome further comprises an oxidized phospholipid, such as OxPAPC. In some embodiments, the liposome comprises OxPAPC, which is an oxidized phospholipid containing fragmented oxygenated sn-2 residues, an oxidized phospholipid containing full-length oxygenated sn-2 residues, and / or an oxidized phospholipid containing five-carbon sn-2 residues with ω-aldehyde or ω-carboxyl groups. In some embodiments, the liposome comprises OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC, or OxPAPC is a phospholipid containing epoxyisoprostanoids.In some embodiments, the liposome comprises an OxPAPC selected from the group consisting of 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxycyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxyisoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine ( In some embodiments, the liposome comprises PGPC; 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, OxPAPC within the lipid bilayer of the liposome is 0%-100% of the total lipids or any range therebetween. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest. In some embodiments, the targeting moiety is attached to one or both of PEG and the outside of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the outside of the liposome by a covalent bond. In some embodiments, the targeting moiety is a polypeptide. In a further embodiment, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the liposome further comprises an immunostimulant (such as 1,6-β-glucan). In some embodiments, the liposome comprises a steric stabilizer. In some embodiments, the steric stabilizer is polyethylene glycol (ie, the liposome is pegylated). In some embodiments, the PEG has a number average molecular weight (Mn) of 200 to 5000 daltons. In other embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential less than or equal to zero. In some embodiments, the liposomes have a zeta potential of -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween.In other embodiments, the liposome is cationic. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero. In some embodiments, the liposome has a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween.

[0413] In some embodiments, the pharmaceutical composition comprises a liposome-encapsulated trans-crocetin salt having the formula: Q-trans-crocetin-Q

[0414]

[0415] in,

[0416] Q is (a) a multivalent counterion or (b) a monovalent cation.

[0417] In some embodiments, Q is a multivalent cationic counterion. In some embodiments, Q is a multivalent metal cation. In further embodiments, Q is a multivalent transition metal cation. In some embodiments, Q is a divalent cationic counterion. In further embodiments, Q is a divalent metal cation. In some embodiments, Q is selected from Ca 2+ Mg 2 + 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ In a further embodiment, Q is Ca 2+ or Mg 2+ In some embodiments, Q is Ca 2+ In some embodiments, Q is Mg 2+ In some embodiments, Q is a divalent organic counterion. In other embodiments, Q is a trivalent cationic counterion, such as Fe 3+ In other embodiments, Q is a multivalent organic counterion. In some embodiments, Q is a divalent organic cation. In some embodiments, Q is a divalent organic cation, such as a protonated diamine.

[0418] In further embodiments, Q is a monovalent cationic counterion. In some embodiments, Q is a monovalent metal cation. In some embodiments, Q is selected from Na + , Li + or K +In some embodiments, Q is an organic cation. In some embodiments, Q is a monovalent organic cation, such as a protonated amine (e.g., a protonated diamine or a protonated polyamine). In some embodiments, Q is an organic cation, such as NH4 + , protonated diamines or protonated polyamines.

[0419] In some embodiments, the liposome contains less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000 or less than 10,000 trans-crocetin molecules. In some embodiments, the liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween trans-crocetin molecules. In some embodiments, the trans-crocetin / lipid ratio of the liposome composition is 1 g / mol and about 1000 g / mol or any range therebetween. In some embodiments, the trans-crocetin / lipid ratio is 10-150 g / mol, 10-100 g / mol, 30-200 g / mol, 40-200 g / mol, or 50-200 g / mol, or any range therebetween. In some embodiments, the liposomes comprise at least 0.1% to 97% trans-crocetin. In some embodiments, the liposomes have a diameter of 20 nm to 500 nm, or 20 nm to 200 nm, or any range therebetween. In some embodiments, the liposomes have 80nm to 120nm, or a diameter of any range therebetween. In some embodiments, the liposome is formed by a liposome component. In a further embodiment, the liposome component comprises at least one of anionic lipids and neutral lipids. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol PEG, and cholesterol-maleimide. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, cholesterol, and HSPC. In another embodiment, the liposome further comprises an oxidized phospholipid, such as OxPAPC. In some embodiments, the liposome comprises OxPAPC, which is an oxidized phospholipid containing a fragmented oxygenated sn-2 residue, an oxidized phospholipid containing a full-length oxygenated sn-2 residue, and / or an oxidized phospholipid containing a five-carbon sn-2 residue with an ω-aldehyde or ω-carboxyl group. In some embodiments, the liposome comprises an OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC, or the OxPAPC is a phospholipid containing epoxyisoprostaglandin. In some embodiments, the liposome comprises an OxPAPC selected from the group consisting of 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxycyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxyisoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine ( In some embodiments, the liposome comprises PGPC; 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, OxPAPC within the lipid bilayer of the liposome is 0%-100% of the total lipids or any range therebetween. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest.In some embodiments, the targeting moiety is attached to one or both of PEG and the outside of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the outside of the liposome by a covalent bond. In some embodiments, the targeting moiety is a polypeptide. In a further embodiment, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the liposome further comprises an immunostimulant (such as 1,6-β-glucan). In some embodiments, the liposome comprises a steric stabilizer. In some embodiments, the steric stabilizer is polyethylene glycol (ie, the liposome is pegylated). In some embodiments, the PEG has a number average molecular weight (Mn) of 200 to 5000 daltons. In other embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential less than or equal to zero. In some embodiments, the liposomes have a zeta potential of -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween. In other embodiments, the liposomes are cationic. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero. In some embodiments, the liposomes have a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween.

[0420] In some embodiments, the present disclosure provides a pharmaceutical composition comprising trans-calcium crocetin (CTC) encapsulated by liposomes. The CTC can exist in linear and / or cyclic forms (as shown below).

[0421]

[0422] In some embodiments, the liposome contains less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000 or less than 10,000 trans-crocetin molecules. In some embodiments, the liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween trans-crocetin molecules. In some embodiments, the trans-crocetin / lipid ratio of the liposome composition is 1 g / mol and about 1000 g / mol or any range therebetween. In some embodiments, the trans-crocetin / lipid ratio is 10-150 g / mol, 10-100 g / mol, 30-200 g / mol, 40-200 g / mol, or 50-200 g / mol, or any range therebetween. In some embodiments, the liposome comprises at least 0.1% to 97% trans-crocetin. In some embodiments, the liposome has a diameter of 20nm to 500nm, or 20nm to 200nm, or any range therebetween. In some embodiments, the liposome has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the liposome is formed by a liposome component. In a further embodiment, the liposome component comprises at least one of anionic lipids and neutral lipids. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol PEG, and cholesterol-maleimide. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, cholesterol, and HSPC. In another embodiment, the liposome further comprises an oxidized phospholipid, such as OxPAPC. In some embodiments, the liposome comprises OxPAPC, which is an oxidized phospholipid containing a fragmented oxygenated sn-2 residue, an oxidized phospholipid containing a full-length oxygenated sn-2 residue, and / or an oxidized phospholipid containing a five-carbon sn-2 residue with an ω-aldehyde or ω-carboxyl group. In some embodiments, the liposome comprises OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC, and KOOA-PC, or OxPAPC is a phospholipid containing epoxyisoprostane.In some embodiments, the liposome comprises an OxPAPC selected from the group consisting of 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxycyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxyisoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine ( In some embodiments, the liposome comprises PGPC; 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, OxPAPC within the lipid bilayer of the liposome is 0%-100% of the total lipids or any range therebetween. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest. In some embodiments, the targeting moiety is attached to one or both of PEG and the outside of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the outside of the liposome by a covalent bond. In some embodiments, the targeting moiety is a polypeptide. In a further embodiment, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the liposome contains less than 500,000 or less than 200,000 trans-crocetin molecules. In some embodiments, the liposome contains between 10 to 100,000 or any range of trans-crocetin molecules therebetween. In some embodiments, the liposome further comprises an immunostimulant (such as 1,6-β-glucan). In some embodiments, the liposome comprises a steric stabilizer. In some embodiments, the steric stabilizer is polyethylene glycol (i.e., the liposome is PEGylated). In some embodiments, the PEG has a number average molecular weight (Mn) of 200 to 5000 Daltons. In further embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential less than or equal to zero.In some embodiments, the liposomes have a zeta potential of -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween. In other embodiments, the liposomes are cationic. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero. In some embodiments, the liposomes have a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween.

[0423] In some embodiments, the present disclosure provides a pharmaceutical composition comprising trans-magnesium crocetin (MTC) encapsulated by liposomes. The MTC can exist in linear and / or cyclic forms (as shown below).

[0424]

[0425] In some embodiments, the liposomes contain less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, or less than 10,000 trans-crocetin molecules. In some embodiments, the liposomes contain 10 to 100,000, 100 to 10,000, or 500 to 5,000, or any range therebetween, trans-crocetin molecules. In some embodiments, the trans-crocetin / lipid ratio is 10-150 g / mol, 10-100 g / mol, 30-200 g / mol, 40-200 g / mol, or 50-200 g / mol, or any range therebetween. In some embodiments, the liposomes contain at least 0.1% to 97% trans-crocetin. In some embodiments, the liposome has a diameter of 20nm to 500nm, or 20nm to 200nm, or any range therebetween. In some embodiments, the liposome has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the liposome is formed by a liposome component. In a further embodiment, the liposome component comprises at least one of anionic lipids and neutral lipids. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol PEG, and cholesterol-maleimide. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, cholesterol, and HSPC. In another embodiment, the liposome further comprises an oxidized phospholipid, such as OxPAPC. In some embodiments, the liposome comprises OxPAPC, which is an oxidized phospholipid containing a fragmented oxygenated sn-2 residue, an oxidized phospholipid containing a full-length oxygenated sn-2 residue, and / or an oxidized phospholipid containing a five-carbon sn-2 residue with an ω-aldehyde or ω-carboxyl group. In some embodiments, the liposome comprises OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC, and KOOA-PC, or OxPAPC is a phospholipid containing epoxyisoprostane.In some embodiments, the liposome comprises an OxPAPC selected from the group consisting of 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxycyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxyisoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine ( In some embodiments, the liposome comprises PGPC; 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, OxPAPC within the lipid bilayer of the liposome is 0%-100% of the total lipids or any range therebetween. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest. In some embodiments, the targeting moiety is attached to one or both of PEG and the outside of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the outside of the liposome by a covalent bond. In some embodiments, the targeting moiety is a polypeptide. In a further embodiment, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the liposome further comprises an immunostimulant (such as 1,6-β-glucan). In some embodiments, the liposome comprises a steric stabilizer. In some embodiments, the steric stabilizer is polyethylene glycol (ie, the liposome is pegylated). In some embodiments, the PEG has a number average molecular weight (Mn) of 200 to 5000 daltons. In other embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential less than or equal to zero. In some embodiments, the liposomes have a zeta potential of -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween.In other embodiments, the liposome is cationic. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero. In some embodiments, the liposome has a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween.

[0426] In some embodiments, the present disclosure provides a pharmaceutical composition comprising trans-norbixin having the formula: Q-norbixin-Q

[0427]

[0428] in,

[0429] Q is a multivalent cationic counterion.

[0430] In some embodiments, Q is a multivalent cationic counterion. In some embodiments, Q is a multivalent metal cation. In further embodiments, Q is a multivalent transition metal cation. In some embodiments, Q is a divalent cationic counterion. In further embodiments, Q is a divalent metal cation. In some embodiments, Q is selected from Ca 2+ Mg 2 + 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ In a further embodiment, Q is Ca 2+ or Mg 2+ In some embodiments, Q is Ca 2+ In some embodiments, Q is Mg 2+ In some embodiments, Q is a divalent organic counterion.

[0431] In some embodiments, Q is a monovalent cationic counterion. In some embodiments, Q is a monovalent metal cation. In some embodiments, Q is selected from Na + , Li + or K + In some embodiments, Q is an organic cation. In some embodiments, Q is a monovalent organic cation, such as a protonated amine (e.g., a protonated diamine or a protonated polyamine). In some embodiments, Q is an organic cation, such as NH4 + , protonated diamines or protonated polyamines.

[0432] In some embodiments, the liposome contains less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, or less than 10,000 trans-norbixin molecules. In some embodiments, the liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-norbixin molecules. In some embodiments, the trans-norbixin / lipid ratio of the liposome composition is 1 g / mol and about 1000 g / mol, or any range therebetween. In some embodiments, the trans-norbixin / lipid ratio is 10-150 g / mol, 10-100 g / mol, 30-200 g / mol, 40-200 g / mol, or 50-200 g / mol, or any range therebetween. In some embodiments, the liposome comprises at least 0.1% to 97% Q-norbixin-Q. In some embodiments, the liposome has a diameter of 20nm to 500nm, or 20nm to 200nm, or any range therebetween. In some embodiments, the liposome has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the liposome is formed by a liposome component. In a further embodiment, the liposome component comprises at least one of anionic lipids and neutral lipids. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol PEG, and cholesterol-maleimide. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, cholesterol, and HSPC. In other embodiments, the liposome further comprises an oxidized phospholipid, such as OxPAPC. In some embodiments, the liposome comprises OxPAPC, which is an oxidized phospholipid containing fragmented oxygenated sn-2 residues, an oxidized phospholipid containing full-length oxygenated sn-2 residues, and / or an oxidized phospholipid containing five-carbon sn-2 residues with ω-aldehyde or ω-carboxyl groups. In some embodiments, the liposome comprises OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC, or OxPAPC is a phospholipid containing epoxyisoprostanoids.In some embodiments, the liposome comprises an OxPAPC selected from the group consisting of 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxycyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxyisoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine ( In some embodiments, the liposome comprises PGPC; 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, OxPAPC within the lipid bilayer of the liposome is 0%-100% of the total lipids or any range therebetween. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest. In some embodiments, the targeting moiety is attached to one or both of PEG and the outside of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the outside of the liposome by a covalent bond. In some embodiments, the targeting moiety is a polypeptide. In a further embodiment, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the liposome further comprises an immunostimulant (such as 1,6-β-glucan). In some embodiments, the liposome comprises a steric stabilizer. In some embodiments, the steric stabilizer is polyethylene glycol (ie, the liposome is pegylated). In some embodiments, the PEG has a number average molecular weight (Mn) of 200 to 5000 daltons. In other embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential less than or equal to zero. In some embodiments, the liposomes have a zeta potential of -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween.In other embodiments, the liposome is cationic. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero. In some embodiments, the liposome has a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween.

[0433] In some embodiments, the present disclosure provides a pharmaceutical composition comprising trans-calcium norbixin (CTN) encapsulated by liposomes. The CTN can exist in linear and / or cyclic forms (as shown below).

[0434]

[0435] In some embodiments, the liposomes contain less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, or less than 10,000 trans-norbixin molecules. In some embodiments, the liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-norbixin molecules. In some embodiments, the trans-norbixin / lipid ratio of the liposome composition is 1 g / mol and about 1000 g / mol, or any range therebetween. In some embodiments, the trans-norbixin / lipid ratio is 10-150 g / mol, 10-100 g / mol, 30-200 g / mol, 40-200 g / mol, or 50-200 g / mol, or any range therebetween. In some embodiments, the liposome comprises at least 0.1% to 97% trans-norbixin. In some embodiments, the liposome has a diameter of 20nm to 500nm, or 20nm to 200nm, or any range therebetween. In some embodiments, the liposome has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the liposome is formed by a liposome component. In a further embodiment, the liposome component comprises at least one of anionic lipids and neutral lipids. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol PEG, and cholesterol-maleimide. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, cholesterol, and HSPC. In another embodiment, the liposome further comprises an oxidized phospholipid, such as OxPAPC. In some embodiments, the liposome comprises OxPAPC, which is an oxidized phospholipid containing a fragmented oxygenated sn-2 residue, an oxidized phospholipid containing a full-length oxygenated sn-2 residue, and / or an oxidized phospholipid containing a five-carbon sn-2 residue with an ω-aldehyde or ω-carboxyl group. In some embodiments, the liposome comprises OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC, and KOOA-PC, or OxPAPC is a phospholipid containing epoxyisoprostane.In some embodiments, the liposome comprises an OxPAPC selected from the group consisting of 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxycyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxyisoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine ( In some embodiments, the liposome comprises PGPC; 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, OxPAPC within the lipid bilayer of the liposome is 0%-100% of the total lipids or any range therebetween. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest. In some embodiments, the targeting moiety is attached to one or both of PEG and the outside of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the outside of the liposome by a covalent bond. In some embodiments, the targeting moiety is a polypeptide. In a further embodiment, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the liposome further comprises an immunostimulant (such as 1,6-β-glucan). In some embodiments, the liposome comprises a steric stabilizer. In some embodiments, the steric stabilizer is polyethylene glycol (ie, the liposome is pegylated). In some embodiments, the PEG has a number average molecular weight (Mn) of 200 to 5000 daltons. In other embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential less than or equal to zero. In some embodiments, the liposomes have a zeta potential of -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween.In other embodiments, the liposome is cationic. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero. In some embodiments, the liposome has a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween.

[0436] In some embodiments, the present disclosure provides a pharmaceutical composition comprising trans-magnesium norbixin (MTN) encapsulated by liposomes. The MTN can exist in linear and / or cyclic forms (as shown below).

[0437]

[0438] In some embodiments, the liposomes contain less than 6 million, less than 500,000, less than 200,000, less than 100,000, less than 50,000, or less than 10,000 trans-norbixin molecules. In some embodiments, the liposomes contain 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, or any range therebetween, trans-norbixin molecules. In some embodiments, the trans-norbixin / lipid ratio of the liposome composition is 1 g / mol and about 1000 g / mol, or any range therebetween. In some embodiments, the trans-norbixin / lipid ratio is 10-150 g / mol, 10-100 g / mol, 30-200 g / mol, 40-200 g / mol, or 50-200 g / mol, or any range therebetween. In some embodiments, the liposome comprises at least 0.1% to 97% trans-norbixin. In some embodiments, the liposome has a diameter of 20nm to 500nm, or 20nm to 200nm, or any range therebetween. In some embodiments, the liposome has a diameter of 80nm to 120nm, or any range therebetween. In some embodiments, the liposome is formed by a liposome component. In a further embodiment, the liposome component comprises at least one of anionic lipids and neutral lipids. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-maleimide, HSPC, HSPC-PEG, cholesterol, cholesterol PEG, and cholesterol-maleimide. In a further embodiment, the liposome component comprises at least one selected from the following: DSPE, DSPE-PEG, DSPE-PEG-FITC, DSPE-PEG-maleimide, cholesterol, and HSPC. In another embodiment, the liposome further comprises an oxidized phospholipid, such as OxPAPC. In some embodiments, the liposome comprises OxPAPC, which is an oxidized phospholipid containing a fragmented oxygenated sn-2 residue, an oxidized phospholipid containing a full-length oxygenated sn-2 residue, and / or an oxidized phospholipid containing a five-carbon sn-2 residue with an ω-aldehyde or ω-carboxyl group. In some embodiments, the liposome comprises OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC, and KOOA-PC, or OxPAPC is a phospholipid containing epoxyisoprostane.In some embodiments, the liposome comprises an OxPAPC selected from the group consisting of 1-palmitoyl-2-(5,6-epoxyisoprostaglandin E2)-sn-glycero-3-phosphocholine (5,6PEIPC), 1-palmitoyl-2-(epoxycyclopentenone)-sn-glycero-3-phosphocholine (PECPC), 1-palmitoyl-2-(epoxyisoprostaglandin E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine ( In some embodiments, the liposome comprises PGPC; 1-palmitoyl-2-(9'-oxo-nonanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelayl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, OxPAPC within the lipid bilayer of the liposome is 0%-100% of the total lipids or any range therebetween. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest. In some embodiments, the targeting moiety is attached to one or both of PEG and the outside of the liposome, optionally wherein the targeting moiety is attached to one or both of PEG and the outside of the liposome by a covalent bond. In some embodiments, the targeting moiety is a polypeptide. In a further embodiment, the targeting moiety is an antibody or an antigen-binding fragment of an antibody. In some embodiments, the liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200, or any range of targeting moieties therebetween. In some embodiments, the liposome contains less than 500,000 or less than 200,000 trans-norbixin molecules. In some embodiments, the liposome contains between 10 to 100,000 or any range of trans-norbixin molecules therebetween. In some embodiments, the liposome further comprises an immunostimulant (such as 1,6-β-glucan). In some embodiments, the liposome comprises a steric stabilizer. In some embodiments, the steric stabilizer is polyethylene glycol (i.e., the liposome is PEGylated). In some embodiments, the PEG has a number average molecular weight (Mn) of 200 to 5000 Daltons. In further embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential less than or equal to zero.In some embodiments, the liposomes have a zeta potential of -150 to 0, -50 to 0 mV, -40 to 0 mV, -30 to 0 mV, -25 to 0 mV, -20 to 0 mV, -10 to 0 mV, -9 to 0 mV, -8 to 0 mV, -7 to 0 mV, -6 to 0 mV, -5 to 0 mV, -4 to 0 mV, -3 to 0 mV, -2 to 0 mV, -1 to 0 mV, or -8 to 2 mV, or any range therebetween. In other embodiments, the liposomes are cationic. In some embodiments, the liposome composition comprises liposomes having a zeta potential greater than zero. In some embodiments, the liposomes have a zeta potential of 0.2 to 150 mV, 1 to 50 mV, 1 to 40 mV, 1 to 30 mV, 1 to 25 mV, 1 to 20 mV, 1 to 15 mV, 1 to 10 mV, 1 to 5 mV, 2 to 10 mV, 3 to 10 mV, 4 to 10 mV, or 5 to 10 mV, or any range therebetween.

[0439] Preparation and administration

[0440] The provided compositions can be formulated as pharmaceutical compositions in whole or in part. Pharmaceutical compositions can include one or more nanoparticle compositions. For example, a pharmaceutical composition can include one or more nanoparticle compositions that include one or more different therapeutic and / or prophylactic drugs. The pharmaceutical composition can further include one or more pharmaceutically acceptable excipients or adjuvants, such as those described herein. General guidelines for the preparation and manufacture of pharmaceutical compositions and agents can be obtained, for example, in Remington's The Science and Practice of Pharmacy, 21st edition, A. R. Gennaro; Lippincott, Williams and Wilkins, Baltimore, Md., 2006. Conventional excipients and adjuvants can be used in any pharmaceutical composition, except where any conventional excipient or adjuvant may be incompatible with one or more components of the nanoparticle composition. If the combination of an excipient or adjuvant with a component of a nanoparticle composition may result in any undesirable biological effect or otherwise harmful effect, the excipient or adjuvant may be incompatible with the component of the nanoparticle composition.

[0441] In some embodiments, one or more excipients or auxiliary ingredients may account for more than 50% of the total mass or volume of the pharmaceutical composition including the nanoparticle composition. For example, the one or more excipients or auxiliary ingredients may account for 50%, 60%, 70%, 80%, 90% or more of the pharmaceutical practice. In some embodiments, the pharmaceutically acceptable excipient is at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% pure. In some embodiments, the excipient is approved for human and veterinary use. In some embodiments, the excipient is approved by the U.S. Food and Drug Administration. In some embodiments, the excipient is pharmaceutical grade. In some embodiments, the excipient meets the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia and / or the International Pharmacopoeia.

[0442] Standard methods for preparing liposomes include, but are not limited to, those reported in Liposomes: A Practical Approach, VP Torchilin, Volkmar Weissig Oxford University Press, 2003, and are well known in the art.

[0443] In some embodiments, the disclosure provides liposome compositions and physiologically (i.e., pharmaceutically) acceptable carriers. As used herein, the term "carrier" refers to a generally inert substance used as a diluent or vehicle for a drug such as a therapeutic agent. The term also encompasses generally inert substances that impart cohesiveness to the composition. Generally, the physiologically acceptable carrier exists in liquid form. Examples of liquid carriers include physiological saline, phosphate buffer, physiological buffered saline (135-150mM NaCl), water, buffered water, 0.4% saline, 0.3% glycine, glycoproteins (e.g., albumin, lipoprotein, globulin, etc.) for providing enhanced stability, etc. Because physiologically acceptable carriers are determined in part by the specific composition being administered and by the specific method for administering the composition, the pharmaceutical composition provided herein has a variety of suitable preparations (see, e.g., Remington's Pharmaceutical Sciences, 17th edition, 1989).

[0444] The provided compositions can be sterilized by conventional, known sterilization techniques or can be produced under aseptic conditions. The aqueous solution can be packaged for use or filtered and lyophilized under aseptic conditions, and the lyophilized preparation is combined with the sterile aqueous solution before administration. The composition can contain pharmaceutically acceptable auxiliary substances as needed to approach physiological conditions, such as pH adjustment and buffering agents, tonicity regulators, wetting agents, etc., for example, sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate and triethanolamine oleate. Sugars for stabilizing the composition can also be included, such as stabilizers for freeze-dried liposome compositions. In some embodiments, the pharmaceutical composition comprises a tonicity agent at a concentration greater than 0.1% or a concentration of 0.3% to 2.5%, 0.5% to 2.0%, 0.5% to 1.5%, 0.5% to 1.5%, 0.6% to 1.1% or any range therebetween. In some embodiments, the pharmaceutical composition comprises a tonicity agent such as dextrose, mannitol, glycerol, potassium chloride or sodium chloride. In further embodiments, the pharmaceutical composition comprises dextrose, mannitol, glycerol, potassium chloride, or sodium chloride at a concentration greater than 0.1% or at a concentration of 0.3% to 2.5%, 0.5% to 2.0%, 0.5% to 1.5%, 0.5% to 1.5%, 0.6% to 1.1%, or any range therebetween.

[0445] Preparations suitable for parenteral administration, such as, for example, administration by intraarticular (in a joint), intravenous, intramuscular, intratumoral, intradermal, intraperitoneal and subcutaneous routes include aqueous and non-aqueous isotonic sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, and solutes that make the preparation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions, which may contain suspending agents, solubilizers, thickeners, stabilizers and preservatives. Injection solutions and suspensions can also be prepared from sterile powders, granules and tablets. In some embodiments, the liposome compositions provided can be administered, for example, by intravenous infusion, topical, intraperitoneal, intravesical or intrathecal administration. In specific embodiments, the liposome compositions are administered parenterally or intravenously. Preferably, the pharmaceutical liposome compositions are administered parenterally, i.e., intraarticularly, intravenously, subcutaneously or intramuscularly. In other embodiments, the pharmaceutical preparation can be administered topically.

[0446] In some embodiments, provided pharmaceutical compositions (eg, liposomal compositions) are presented in unit-dose or multi-dose sealed containers, such as ampoules and vials.

[0447] In some embodiments, the pharmaceutical preparation is administered in unit dosage form. In such forms, the preparation is subdivided into unit doses containing an appropriate amount of active ingredient (e.g., liposome composition). The unit dosage form can be a packaged preparation containing a discrete amount of the preparation. If desired, the composition can also contain other compatible therapeutic agents (e.g., as described herein).

[0448] In some embodiments, the liposome composition comprising the therapeutic agent and / or diagnostic agent used in the pharmaceutical composition provided herein can be administered at an initial dose of about 0.001 mg / kg to about 1000 mg / kg per day. A daily dosage range of about 0.01 mg / kg to about 500 mg / kg or about 0.1 mg / kg to about 200 mg / kg or about 1 mg / kg to about 100 mg / kg or about 10 mg / kg to about 50 mg / kg can be used. However, the dosage can vary according to the needs of the patient, the severity of the condition being treated, and the liposome composition being used. For example, the type and stage of the disease, illness, or condition diagnosed in a particular patient can be considered and the dosage is determined empirically. The dosage administered to the patient should be sufficient to affect the beneficial therapeutic response of the patient over time in the context of the pharmaceutical composition provided (e.g., liposome composition). The size of the dosage will also be determined by the presence, nature, and degree of any adverse side effects associated with the specific liposome composition when it is administered in a specific patient. The determination of the appropriate dosage for a particular situation is within the technical scope of the practitioner. Generally, treatment is started with a smaller dose that is lower than the optimal dose of the liposome composition. Thereafter, the dose is increased in small increments until the optimal effect under the circumstances is reached. For convenience, the total daily dose may be divided and administered in several portions during the day as needed.

[0449] Liposome loading

[0450] The provided carotenoid compositions can be loaded into liposomes using active or passive loading methods.

[0451] In some embodiments, the present disclosure provides a method for preparing a liposome composition comprising ionizable crocetin (e.g., the crocetin described in [1]-

[97] ), the method comprising:

[0452] (a) forming a mixture comprising: a liposome component in a solution;

[0453] (b) homogenizing the mixture to form liposomes in solution; and

[0454] (c) processing the mixture to form liposomes containing the ionizable carotenoid.

[0455] In some embodiments, the processing step comprises one or more of the following steps: thin film hydration, extrusion, online mixing, ethanol injection technology, freeze-thaw technology, reverse phase evaporation, dynamic high pressure microfluidization, microfluidic mixing, double emulsion, freeze-dried double emulsion, 3D printing, membrane contactor method and stirring. In some embodiments, the processing step comprises one or more steps of changing the size of the liposomes by one or more of extrusion, high pressure microfluidization and / or ultrasound steps.

[0456] In some embodiments, the present disclosure provides methods for generating active loading of carotenoid salts inside liposomal formulations using a soluble acetate metal salt gradient (calcium acetate or magnesium acetate).

[0457] The multivalent counterions used in accordance with the present disclosure can be encapsulated in liposomes according to techniques described herein or otherwise known in the art. This includes passive encapsulation techniques described below or otherwise known in the art.

[0458] In some embodiments, the present disclosure provides a method of preparing a pharmaceutical composition, the method comprising:

[0459] (a) preparing a liposome solution comprising liposomes in a weak acid salt of a multivalent metal;

[0460] (b) adding an ionizable carotenoid to the liposome solution; and

[0461] (c) maintaining the ionizable carotenoid in the liposome solution for a sufficient time to load the carotenoid into the liposomes.

[0462] In some embodiments, the ionizable carotenoid is an ionizable carotenoid of any one of compositions [1]-

[28] (e.g., trans-crocetin and trans-norbixin). In some embodiments, the carotenoid is Figures 1A to 1D In some embodiments, the weak acid is selected from acetic acid, gluconic acid, tartaric acid, glutamic acid, citric acid, formic acid and glycine. In some embodiments, the weak acid salt of the multivalent metal is used in a concentration of 0 mM to 2000 mM, or 50 mM to 500 mM, or any range therebetween. In some embodiments, the multivalent metal is selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ , Fe 2+ and Fe 3+ In some embodiments, the weak acid is acetic acid and the multivalent metal is Ca 2+(i.e., the weak acid salt of the polyvalent metal is calcium acetate). In some embodiments, the weak acid is acetic acid and the polyvalent metal is Mg 2+ (i.e., the weak acid salt of the multivalent metal is magnesium acetate). The present disclosure also encompasses pharmaceutical compositions prepared according to the provided methods. The liposome solution is preferably a buffered solution. However, it should be understood that any suitable solvent can be used to prepare and use the provided compositions. The preferred liposome solution has a pH value of about physiological pH and contains a buffer having a buffer range including physiological pH. A non-limiting example of a suitable buffer for the liposome solution is HEPES (e.g., 5mM HEPES buffered saline pH 6.5). The present disclosure also encompasses pharmaceutical compositions prepared according to the method.

[0463] The polyvalent metal used according to the provided method can be encapsulated in liposomes according to conventional techniques known in the art. These methods include, for example, passive encapsulation techniques described herein or otherwise known in the art. By keeping the ionizable carotenoids in the liposome solution at a suitable temperature for a suitable time, the loading of ionizable carotenoids such as trans-crocetin can be established. Depending on the composition of the liposomes and the temperature, pH value and chemical properties of the ionizable carotenoids, the loading of the ionizable carotenoids can be carried out in a time period of several minutes or hours. In some embodiments, loading is carried out at a temperature of, for example, 0°C to 95°C, or 20°C to 75°C, or any range therebetween (preferably about 40°C to about 80°C, or any range therebetween).

[0464] In some embodiments, the disclosure further provides a step (d) of removing unencapsulated ionizable carotenoids from the liposome preparation prepared according to (c). In some embodiments, the removal is carried out by passing the liposome preparation through a gel filtration column, centrifugation, or dialysis, or related techniques balanced with a second aqueous buffer solution. After removing unencapsulated ionizable carotenoids, the degree of ionizable carotenoid loading can be determined according to conventional techniques by measuring ionizable carotenoids and lipid levels. Lipid and drug concentrations can be measured using any suitable method known in the art, such as scintillation counting, spectrophotometry, and high performance liquid chromatography. Any of a variety of techniques known in the art can be used to complete the replacement of liposome preparation solutions to remove unencapsulated carotenoids and counterions (such as sodium acetate), including but not limited to chromatography of the liposome preparation by an extensive gel filtration column balanced with a second aqueous buffer solution, by centrifugation, extensive or repeated dialysis, replacement of the liposome preparation, treatment of the liposome preparation with a chelating agent, or by related techniques. The present disclosure also encompasses pharmaceutical compositions prepared according to the provided methods.

[0465] In some embodiments, the present disclosure provides a method of preparing a pharmaceutical composition, the method comprising:

[0466] (a) preparing a liposome solution comprising liposomes in a weak acid salt of a multivalent metal;

[0467] (b) adding trans-crocetin to the liposome solution; and

[0468] (c) maintaining the ionizable carotenoid in the liposome solution for a sufficient time to load the carotenoid into the liposomes.

[0469] In some embodiments, the weak acid is selected from acetic acid, gluconic acid, tartaric acid, glutamic acid, citric acid, formic acid and glycine. In some embodiments, the weak acid salt of the polyvalent metal is used in a concentration of 0 mM to 2000 mM, or 50 mM to 500 mM, or any range therebetween. In some embodiments, the polyvalent metal is selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ , Fe 2+ and Fe 3+ In some embodiments, the weak acid is acetic acid and the multivalent metal is Ca 2+ (i.e., the weak acid salt of the polyvalent metal is calcium acetate). In some embodiments, the weak acid is acetic acid and the polyvalent metal is Mg 2+ (i.e., the weak acid salt of the multivalent metal is magnesium acetate). The present disclosure also encompasses pharmaceutical compositions prepared according to the method. The liposome solution is preferably a buffered solution. However, it should be understood that any suitable solvent can be used to practice the provided compositions and methods. The preferred liposome solution has a pH value of about physiological pH and contains a buffer having a buffer range including physiological pH. A non-limiting example of a suitable buffer for the liposome solution is 5mM HEPES buffered saline pH 6.5. The present disclosure also encompasses pharmaceutical compositions prepared according to the method.

[0470] The loading of trans-crocetin can be established by keeping trans-crocetin in the liposome solution at a suitable temperature for a suitable time. Depending on the composition of the liposomes and the temperature, pH and chemical properties of the trans-crocetin, the loading of the trans-crocetin can be carried out over a period of minutes or hours. In some embodiments, the loading is carried out at a temperature of, for example, 0°C to 95°C, or 20°C to 75°C, or any range therebetween, preferably about 40°C to about 80°C.

[0471] In some embodiments, the present disclosure further provides a step (d) of removing unencapsulated trans-crocetin from the liposome preparation prepared according to (c). In some embodiments, the removal is carried out by passing the liposome preparation through a gel filtration column balanced with a second aqueous buffer solution, or by centrifugation, dialysis or related techniques. After removing unencapsulated trans-crocetin, the degree of trans-crocetin loading can be determined by measuring trans-crocetin and lipid levels according to conventional techniques. Lipid and drug concentrations can be determined by any suitable method known in the art, such as scintillation counting, spectrophotometry and high performance liquid chromatography. Any of a variety of techniques known in the art can be used to complete the replacement of the liposome preparation solution to remove unencapsulated trans-crocetin and counterions (such as sodium acetate), including but not limited to chromatography of the liposome preparation by an extensive gel filtration column balanced with a second aqueous buffer solution, by centrifugation, extensive or repeated dialysis, exchange of the liposome preparation, treatment of the liposome preparation with a chelating agent or related techniques. The present disclosure also encompasses pharmaceutical compositions prepared according to the provided methods.

[0472] The present disclosure also encompasses pharmaceutical compositions comprising an ionizable carotenoid salt prepared according to the provided methods. In some embodiments, the ionizable carotenoid is an ionizable carotenoid of any one of compositions [1]-

[28] (e.g., trans-crocetin and trans-norbixin). In some embodiments, the ionizable carotenoid is Figures 1A-1D In some embodiments, the present disclosure provides a pharmaceutical composition comprising a liposome encapsulating an ionizable carotenoid, wherein the ionizable carotenoid is encapsulated by a multivalent counter ion (e.g., Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ and Fe 3+ ) is loaded into the liposome in the presence of. In some embodiments, the multivalent counterion comprises Ca 2+ In some embodiments, the multivalent counterion comprises Mg 2+ In some embodiments, the multivalent counterion comprises Fe 3+ .

[0473] In some embodiments, the present disclosure provides a pharmaceutical composition comprising liposomes encapsulating trans-crocetin, wherein the trans-crocetin is encapsulated within the liposome by a multivalent counterion (e.g., Ca 2+ Mg 2+ 、Zn 2+ , Cu2+ 、Co 2+ and Fe 2 + and Fe 3+ ) is loaded into the liposome in the presence of. In some embodiments, the multivalent counterion comprises Ca 2+ In some embodiments, the multivalent counterion comprises Mg 2+ In some embodiments, the multivalent counterion comprises Fe 3+ .

[0474] Treatment and methods of administration

[0475] The uses of the provided pharmaceutical compositions (such as liposome compositions) are an improvement over previous treatments for diseases and conditions including, but not limited to, infections and infectious diseases such as HIV / AIDS: human immunodeficiency virus-1 (HIV-1), tuberculosis, malaria and its complications (such as cerebral malaria), severe anemia, acidosis, acute renal failure and ARDS, sepsis, inflammation (e.g., chronic inflammatory diseases), ischemia (including ischemic conditions such as ischemic stroke, coronary artery disease, peripheral vascular disease, cerebrovascular disease, ischemia-related renal pathology, and wound-related ischemia); shock (e.g., hemorrhagic shock), stroke, cardiovascular disease, renal pathology, wound healing, metabolic diseases, hyperproliferative diseases (such as cancer), and disorders of the immune system, cardiovascular system, digestive system, nervous system, respiratory system, and endocrine system. In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition in a subject in need of such treatment or prevention, the method comprising administering to the subject a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ). Also provided herein is the use of a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) in the manufacture of a medicament for treating a disease, disorder or condition in a subject. Also provided herein is a pharmaceutical composition described in any one of [1]-

[79] for use in a medical drug.

[0476] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with endotoxemia in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any of [1]-

[79] ) to the subject.

[0477] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with sepsis in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the subject suffers from a low-grade endotoxemia-type disease.

[0478] In some embodiments, the present disclosure provides a method for treating or preventing a subject at risk of developing sepsis, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the subject is immunocompromised or immunosuppressed. In some embodiments, the subject is critically ill. In some embodiments, the subject is an elderly person or a newborn. In some embodiments, the subject has febrile neutropenia. In some embodiments, the subject has an infection.

[0479] In some embodiments, the present disclosure provides a method for treating or preventing a burn-related disease, disorder, or condition in a subject who is a burn victim, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject.

[0480] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with an infection in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the infection is a bacterial infection (e.g., a Pseudomonas aeruginosa infection, a Staphylococcus aureus infection (e.g., MRSA), a Mycobacterium tuberculosis infection, an Enterococcus infection (e.g., VRE) or a condition associated therewith. In some embodiments, the infection is a fungal infection (e.g., a Candidiasis infection, such as invasive Candidiasis) or a condition associated therewith. In some embodiments, the infection is a parasitic infection (e.g., schistosomiasis and human African trypanosomiasis) or a condition associated therewith. In some embodiments, the infection is malaria or a condition associated therewith, such as cerebral malaria, severe anemia, acidosis, acute renal failure, and ARDS. In some embodiments, the infection is a viral infection (e.g., Ebola, dengue fever, and Marburg) or a condition associated therewith, such as influenza, measles, and viral hemorrhagic fever.

[0481] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder or condition associated with ischemia or hypoxia in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the disease or condition associated with ischemia or hypoxia is associated with surgery or traumatic injury. In some embodiments, the disease or condition is ischemia-reperfusion injury, transient cerebral ischemia, cerebral ischemia-reperfusion, ischemic stroke, hemorrhagic stroke, traumatic brain injury, migraine (e.g., chronic migraine or severe migraine), gastrointestinal ischemia, kidney disease, pulmonary embolism, acute respiratory failure, neonatal respiratory distress syndrome, obstetric emergency for reducing perinatal complications (such as, pre-eclampsia / eclampsia and conditions leading to cerebral palsy), myocardial infarction, acute limb or mesenteric ischemia, cardiogenic cirrhosis, chronic peripheral vascular disease, congestive heart failure, atherosclerotic stenosis, anemia, thrombosis, embolism, macular degeneration, neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis (ALS), sleep apnea and surgery or traumatic injury). In some embodiments, the disease or condition associated with ischemia or hypoxia is myocardial infarction or congestive heart failure with or without cardiogenic cirrhosis. In some embodiments, the disease or condition is pulmonary embolism, acute respiratory failure, chronic peripheral vascular disease, atherosclerotic stenosis, anemia, thrombosis or embolism. In some embodiments, the disease or condition associated with ischemia or hypoxia is macular degeneration or an oncological condition associated with hypoxia. In some embodiments, the disease or condition is kidney disease. In some embodiments, the disease or condition is acute kidney injury (AKI) or end-stage renal disease induced by lipopolysaccharide drugs or toxins.

[0482] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with shock in a subject in need of such treatment or prevention, the method comprising administering to the subject a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any of [1]-

[79] ). In some embodiments, the disease or condition is associated with cardiogenic shock. In some embodiments, the disease or condition is associated with hypovolemic shock. In some embodiments, the disease or condition is associated with septic shock or other forms of distributive shock. In some embodiments, the disease or condition is associated with neurogenic shock. In some embodiments, the disease or condition is associated with anaphylactic shock.

[0483] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with nitric oxide deficiency in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the disease or disorder is sickle cell disease, paroxysmal nocturnal hemoglobinuria (PNH), hemolytic anemia, thalassemia, another red blood cell disorder, or a condition associated therewith. In some embodiments, the disease or disorder is purpura, such as thrombotic thrombocytic purpura (TTP), hemolytic uremic syndrome (HUS), idiopathic thrombocytopenia (ITP), or another platelet disorder, or a condition associated therewith. In some embodiments, the disease or disorder is a coagulation disorder, such as disseminated intravascular coagulopathy (DIC), purpura fulminans, heparin-induced thrombocytopenia (HIT), leukocytosis, hyperviscosity syndrome, or a condition associated therewith.

[0484] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder or condition associated with inflammation in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the disease or condition associated with inflammation is low-grade inflammation. In some embodiments, the disease or condition associated with inflammation is systemic inflammation. In some embodiments, the disease or condition associated with inflammation is acute inflammation or a chronic inflammatory disease.

[0485] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder or condition associated with a cardiovascular disease or condition in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the cardiovascular disease or condition is coronary artery disease. In some embodiments, the cardiovascular disease or condition is myocardial infarction, sudden cardiac death, cardiopulmonary arrest, hypertension, pulmonary hypertension, atherosclerosis, occlusive arterial disease, Raynaud's disease, peripheral vascular disease, other vascular diseases such as Buerger's disease, Takayasu's arthritis and post-cardiac arrest syndrome (PCAS), chronic venous insufficiency, heart disease, congestive heart failure, or chronic skin ulcers.

[0486] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder or condition associated with a liver disease or condition in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the liver disease or condition is cirrhosis. In some embodiments, the liver disease or condition is non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH). In some embodiments, the liver disease or condition is alcoholic liver disease. In some embodiments, the liver disease or condition is acute liver injury.

[0487] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder or condition associated with a lung disease or condition in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the lung disease or condition is acute respiratory distress syndrome (ARDS). In some embodiments, the lung disease or condition is chronic obstructive pulmonary disease. In some embodiments, the lung disease or condition is pulmonary fibrosis. In some embodiments, the lung disease or condition is pulmonary hemorrhage. In some embodiments, the lung disease or condition is asthma. In some embodiments, the lung disease or condition is lung injury. In some embodiments, the lung disease or condition is lung cancer. In some embodiments, the condition is cystic fibrosis.

[0488] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder or condition associated with a kidney disease or condition in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the kidney disease or condition is lipopolysaccharide-induced acute kidney injury (AKI). In some embodiments, the kidney disease or condition is chronic renal failure with or without end-stage renal disease.

[0489] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with a vascular disease in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the disease or condition is coronary artery disease. In some embodiments, the disease or condition is hypertension. In some embodiments, the disease or condition is atherosclerosis. In some embodiments, the disease or condition is post-cardiac arrest syndrome (PCAS). In some embodiments, the disease or condition is occlusive arterial disease, peripheral vascular disease, chronic venous insufficiency, chronic skin ulcers, or Raynaud's disease. In some embodiments, the disease, disorder, or condition associated with a vascular disease is heart disease. In further embodiments, the disease, disorder, or condition is congestive heart failure. In some embodiments, the disease, disorder, or condition associated with a vascular disease is ischemic bowel disease.

[0490] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with heart attack or stroke in a subject in need of such treatment or prevention and / or at risk of heart attack or stroke, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any of [1]-

[79] ) to the subject. In some embodiments, the disease, disorder, or condition is ischemic stroke. In some embodiments, the disease, disorder, or condition is hemorrhagic stroke.

[0491] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with the nervous system in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the disease or condition is pain (e.g., chronic pain). In some embodiments, the disease or condition is a neurodegenerative disease (e.g., Alzheimer's disease or Parkinson's disease). In some embodiments, the disease, disorder, or condition associated with the nervous system is nerve damage.

[0492] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with inflammatory bowel disease in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any of [1]-

[79] ) to the subject. In some embodiments, the disease, disorder, or condition is Crohn's disease. In some embodiments, the disease, disorder, or condition is ulcerative colitis.

[0493] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder or condition associated with type 2 diabetes or diabetes susceptibility in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ) to the subject. In some embodiments, the disease, disorder or condition is a metabolic disease. In some embodiments, the disease, disorder or condition is insulin resistance. In some embodiments, the disease, disorder or condition is diabetic vascular disease (e.g., microvascular disease, such as retinopathy and nephropathy). In some embodiments, the disease, disorder or condition is diabetic neuropathy. In some embodiments, the disease, disorder or condition is an ulcer, diabetic necrosis or gangrene.

[0494] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with an autoimmune disorder in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any of [1]-

[79] ) to the subject. In some embodiments, the autoimmune disorder is psoriasis. In some embodiments, the autoimmune disorder is cystic fibrosis. In some embodiments, the autoimmune disorder is rheumatoid arthritis.

[0495] In some embodiments, the present disclosure provides a method for treating or preventing a disease, disorder, or condition associated with sclerosis in a subject in need of such treatment or prevention, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition of any one of [1]-

[79] ) to the subject. In some embodiments, the disease, disorder, or condition associated with sclerosis is systemic sclerosis.

[0496] In some embodiments, the present disclosure provides a method of treating endotoxemia in a subject in need of such treatment, the method comprising administering to the subject a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any one of [1]-

[79] ). In some embodiments, the endotoxemia is associated with a condition such as periodontal disease (e.g., periodontitis or gingival inflammation), chronic alcoholism, chronic smoking, transplantation, or neonatal necrotizing enterocolitis, or neonatal ear infection.

[0497] In some embodiments, the present disclosure provides a method for reducing systemic levels of LPS, endotoxin, and / or another systemic inflammatory trigger in a subject in need thereof, the method comprising administering a pharmaceutical composition provided herein (e.g., a pharmaceutical composition described in any of [1]-

[79] ) to the subject.

[0498] Combination therapy

[0499] The compositions provided herein can be administered alone or together with one or more additional therapeutic agents in combination therapy. In some embodiments, the composition is administered together with another therapeutic agent in combination therapy. The combination can be administered concomitantly (e.g., combined in the same delivery vehicle (e.g., liposomes)), as a mixture, separately but simultaneously or synchronously; or sequentially. This includes presentations in which the combined therapeutic agents are administered together as a therapeutic mixture, and procedures in which the combined agents are administered separately but simultaneously (e.g., via separate intravenous lines into the same individual). "Combination" administration further includes the separate administration of a therapeutic agent first administered, followed by the separate administration of a second therapeutic agent. A method of treating using combination therapy is also provided.

[0500] In other embodiments, the compositions provided herein are administered in combination with another therapeutic agent. In some embodiments, the compositions described in any one of [1]-

[28] are administered in combination with another therapeutic agent. Figures 1A-1D The composition of the salt of carotenoid provided in any one of the embodiments is administered together with another therapeutic agent as a combination therapy. Figures 1A-1D A composition comprising a multivalent salt (e.g., a divalent salt or a trivalent salt) of a carotenoid provided in any one of the embodiments is administered in combination therapy with another therapeutic agent. In a specific embodiment, a composition comprising a multivalent salt of trans-crocetin (e.g., CTC or MTC) is administered in combination therapy with another therapeutic agent. In other specific embodiments, a composition comprising a multivalent salt of trans-norbixin (e.g., CTN or MTN) is administered in combination therapy with another therapeutic agent.

[0501] In some embodiments, a pharmaceutical composition comprising a salt of one or more ionizable carotenoids is administered in combination therapy with a carotenoid comprising at least one polar group or monocyclic group. In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., a salt containing a divalent, trivalent, or tetravalent counterion). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1DCarotenoids described in any one of the above. In one embodiment, the carotenoid comprising at least one polar group or monocyclic group polar group is symmetrical. In another embodiment, the divalent ionizable carotenoid salt composition is administered in combination therapy with at least one carotenoid selected from the following: zeaxanthin, astaxanthin, lutein and lutein. In another embodiment, the divalent ionizable carotenoid salt composition is administered in combination therapy with astaxanthin. In another embodiment, the carotenoid comprising at least one polar group or monocyclic group polar group is asymmetrical. In another embodiment, the divalent ionizable carotenoid salt composition disclosed herein is administered in combination with abscisic acid (ABA).

[0502] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered as a combination therapy with a standard of care treatment for the disease, disorder, or condition to be treated. In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1D In certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0503] In some embodiments, the pharmaceutical composition comprising the ionizable carotenoid salt provided herein is administered in combination therapy with an antimicrobial agent. In some embodiments, the antimicrobial agent is an antibacterial agent. In some embodiments, the antibacterial agent is selected from, but not limited to, ertapenem, piperacillin-tazobactam, cefepime, aztreonam, metronidazole, meropenem, ceftriaxone, ciprofloxacin, vancomycin, linezolid, tobramycin, levofloxacin, azithromycin, cefazolin and ampicillin. In some embodiments, the antibacterial agent is selected from, but not limited to, ceftriaxone, levofloxacin, ciprofloxacin, cefazolin, piperacillin-tazobactam, meropenem, metronidazole, vancomycin and ampicillin. In other embodiments, the antimicrobial agent is an antifungal agent. In further embodiments, the antifungal agent is caspofungin or another antifungal agent. In other embodiments, the antimicrobial agent is an antimalarial agent. In further embodiments, the antimalarial agent is selected from, but not limited to, artemisinin and its analogs, chloroquine and its analogs, atovaquone, a quinine derivative, proguanil, or another antimalarial agent. In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1DIn certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0504] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with activated protein C (e.g., rhAPC) or drotrecogin alfa (activated) (DAA). In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1D In certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0505] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt as provided herein is administered in combination therapy with a corticosteroid (e.g., a glucocorticoid or a mineralocorticoid, such as fludrocortisonel). In some embodiments, the corticosteroid is a glucocorticoid. In further embodiments, the glucocorticoid is selected from cortisone, ethamethasoneb, prednisone, prednisolone, triamcinolone, dexamethasone, and methylprednisolone. In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1D In certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0506] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with a vitamin administered intravenously. In some embodiments, the vitamin is vitamin C (ascorbic acid). In some embodiments, the vitamin is vitamin A. In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1DIn certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0507] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with a glucocorticoid and vitamin C (e.g., intravenous vitamin C administration). In some embodiments, the glucocorticoid is selected from cortisone, ethamethasoneb, prednisone, prednisolone, triamcinolone, dexamethasone, and methylprednisolone. In further embodiments, the glucocorticoid is hydrocortisone. In additional embodiments, at least one ionizable carotenoid composition provided herein (e.g., comprising Figure 1A , Figure 1B , Figure 1C and / or Figure 1D In some embodiments, the ionizable carotenoid is a divalent salt composition of a carotenoid disclosed in [1]-

[28] and / or Figures 1A-1D In certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0508] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with a vasopressor. In some embodiments, the vasopressor agent is norepinephrine or a similar drug or angiotensin II (e.g., GIAPREZA TM ). In some embodiments, the vasopressor agent is epinephrine, phenylephrine, dopamine, or vasopressin. In some embodiments, the vasopressor agent is ephedrine, milrinone, isoproterenol, dobutamine, isoproterenol, or dopamine.

[0509] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with a thrombolytic agent. In some embodiments, the thrombolytic agent is tissue plasminogen activator (tPA). In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1DIn certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0510] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with a therapeutic agent. In some embodiments, a pharmaceutical composition as described in any one of [1]-

[28] is administered in combination with a therapeutic agent. Figures 1A-1D The pharmaceutical composition of a multivalent salt of a carotenoid provided by any one of the above is administered in combination therapy with a therapeutic agent. In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1D In certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0511] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with a therapeutic agent selected from the group consisting of heparin, vasopressin, antidiuretic hormone (ADH), and a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (statin). In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1D In certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0512] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with an anti-inflammatory therapeutic agent. In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1D In certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0513] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with oxygen and / or intravenous fluids or hyperbaric therapy for maintaining / increasing blood oxygen levels and / or blood pressure. In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1D In certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0514] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with a chemotherapeutic agent (e.g., to enhance the effect of chemotherapy on cancer cells and mitigate the effects of chemotherapy-induced bone marrow suppression and anemia). In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1D In certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0515] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with an immunotherapy. In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1D In certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0516] In some embodiments, a pharmaceutical composition comprising an ionizable carotenoid salt provided herein is administered in combination therapy with radiation therapy. In some embodiments, the salt of the ionizable carotenoid is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the ionizable carotenoid is [1]-

[28] and / or Figures 1A-1DIn certain embodiments, the ionizable carotenoid is trans-crocetin (e.g., CTC and MTC). In other specific embodiments, the ionizable carotenoid is trans-norbixin (e.g., CTN and MTN).

[0517] Kits for administration of active agents

[0518] In another embodiment, the present disclosure provides a kit for administering the provided ionizable carotenoid composition to a subject to treat a disease, disorder or condition. In some embodiments, the present disclosure provides a kit for delivering a therapeutic agent to a subject, the kit comprising: (a) a first composition comprising the disclosed ionizable carotenoid composition (e.g., a liposome comprising a multivalent trans-crocetate); and (b) a second composition containing, for example, a reagent, a buffer, an excipient, or another therapeutic agent stored separately before being administered to a subject. Such kits typically include two or more components necessary for treating disease states such as hypoxia or inflammation-related conditions. In some embodiments, the kit includes, for example, a lipid composition, a reagent, a buffer, a container and / or an apparatus provided. The liposome composition and formulation may be in lyophilized form and then reconstituted before administration. In some embodiments, the kit includes a packaging assembly including one or more components for treating a patient's disease state. For example, a packaging assembly may include a separate container that holds therapeutic liposomes and other excipients or therapeutic agents that may be mixed with the composition before being administered to a patient. In some embodiments, a physician may select and match certain components and / or packaging assemblies, depending on the treatment or diagnosis desired for a particular patient.

[0519] Example

[0520] Example 1 - Production of trans-calcium crocetin liposomes

[0521] Trans-crocetin liposomes were produced using two different variants of trans-crocetin, namely: trans-crocetin free acid (TC) and its sodium salt, trans-sodium crocetin (STC). Trans-crocetin was encapsulated in liposomes by the following procedure.

[0522] Multilayer vesicle (MLV) production :

[0523] First, the lipid components of the liposome lipid membrane are weighed out and combined in the form of a concentrated solution in ethanol at a temperature of about 65° C. In one preparation, the lipids used are hydrogenated soybean phosphatidylcholine, cholesterol, and DSPE-PEG-2000 (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000]). The molar ratio of HSPC: cholesterol: PEG-DSPE is approximately 3:2:0.15. In another preparation, the lipids used are HSPC, cholesterol, PEG-DSPE-2000, and 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine (PGPC). The molar ratio of HSPC: cholesterol: PEG-DSPE: PGPC is approximately 2.7:2:0.15:0.3. Next, calcium acetate is dissolved in an aqueous buffer at a concentration of 125 mM or 250 mM, with a pH of 7.0. Heat the calcium acetate solution to 65°C.

[0524] The ethanol lipid solution was added to the calcium acetate solution using a pipette. In this step, the solution was stirred thoroughly using a magnetic stirrer. The mixing was performed at elevated temperatures (63°C-72°C) to ensure that the lipids were in a liquid crystal state (as opposed to the gel state they would reach at temperatures below the lipid transition temperature (Tm=51°C-54°C). As a result, the lipids were hydrated and formed into multiple bilayer (multilayer) vesicles (MLVs) containing calcium acetate in the interior space.

[0525] Use filter extrude to reduce the size of the MLV:

[0526] MLV fragmentation is made into unilamellar (single bilayer) vesicles of desired size by high pressure extrusion through stacked (track-etched polycarbonate) membrane twice. Stacked membrane has two layers with pore size of 200nm and six layers with pore size of 100nm. During extrusion, the temperature is kept above Tm to ensure the plasticity of lipid membrane. As a result of this extrusion, large and uneven MLVs of size and lamellarity become small uniform (100-120nm) unilamellar vesicles (ULV), which isolate calcium acetate in its internal space. The hydrodynamic size (diameter) of vesicles is measured at 25°C in plastic micro cuvettes using a Malvern Zetasizer Nano ZS instrument (Southborough, MA) with a backscattering detector (90°). Prior to analysis, samples are diluted 50 times in the formulation matrix.

[0527] After the production of ULVs containing calcium acetate, calcium acetate outside the liposomes is removed using SEC (size exclusion chromatography with a PD-10 column) or TFF (tangential flow diafiltration). A tonicity agent is added to the liposomes to balance the osmotic pressure (final concentration: 5% dextrose for 125 mM calcium acetate liposomes and 10% dextrose for 250 mM calcium acetate liposomes). Once the calcium acetate gradient is generated, the trans-crocetin loading procedure is preferably performed within 24 hours. The lipid content of the prepared liposome solution is determined by phosphate assay.

[0528] A 1 mg / mL trans-crocetin solution was prepared in 10% dextrose (for 250 mM calcium acetate liposomes) and the pH was adjusted to 8. The trans-crocetin solution was mixed with the calcium acetate liposome solution at different drug / lipid ratios (100 g / mM, 80 g / mM, 60 g / mM or 40 g / mM). The mixture was then stirred thoroughly and heated to 65°C for 30 minutes and then rapidly cooled to room temperature using an ice-water bath. This step can be replaced by stirring the mixture overnight at room temperature.

[0529] The movement of trans-crocetin molecules (uncharged, neutral form) through the liposome lipid bilayer is driven by a gradient created using calcium acetate (in other words, acetic acid diffuses out and trans-crocetin diffuses in). Trans-crocetin is then entrapped inside the liposomes by ionization and subsequent formation of a calcium-containing precipitate as a calcium salt (trans-crocetin calcium, CTC).

[0530] Purification of liposomes:

[0531] Trans-crocetin outside the liposome was removed using SEC (PD-10 column) or TFF. In this embodiment, the buffer used in SEC was HBS (HEPES buffered saline, pH 6.5). After purification, filtration sterilization was performed using a 0.22 micron filter. The hydrodynamic size (diameter) of the vesicles was measured at 25°C in a plastic micro cuvette using a Malvern Zetasizer Nano ZS instrument (Southborough, MA) with a backscatter detector (90°). The samples were diluted before analysis.

[0532] Table 1: Physical properties of representative CTC-loaded nanoparticles

[0533]

[0534] Example 2 - Using Preparation of calcium acetate liposomes

[0535] Liposomes loaded with calcium acetate were prepared by the following procedure. First, the lipid components of the liposome lipid membrane were weighed out and combined as a concentrated solution in ethanol at a temperature of about 65° C. In one example, the lipids used were hydrogenated soybean phosphatidylcholine, cholesterol, and DSPE-PEG-2000 (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy-(polyethylene glycol)-2000]).

[0536] The molar ratio of HSPC: cholesterol: PEG-DSPE is about 3: 2: 0.15. In another example, the lipids used are HSPC, cholesterol, PEG-DSPE-2000 and 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine (PGPC). The molar ratio of HSPC: cholesterol: PEG-DSPE: PGPC is about 2.7: 2: 0.15: 0.3.

[0537] Next, calcium acetate was dissolved in an aqueous buffer at a concentration of 125 or 250 mM, pH 7.0. The calcium acetate solution was heated to 65°C. The ethanol lipid solution and the calcium acetate solution were transferred to syringes separately. Both solutions were injected into the microfluidic channel and injected with a Precision NanoSystems The device mixes the two solutions as they flow through the microfluidic channel. The mixing is performed at an elevated temperature (63°C-72°C) to ensure that the lipids are in a liquid crystal state (as opposed to the gel state they would reach at temperatures below the lipid transition temperature (Tm=51°C-54°C)). The size of the liposomes can be controlled by the ratio between the lipid solution and the aqueous solution and the mixing flow rate.

[0538] Example 3 - MTC liposome production and characterization

[0539] Production of trans-crocetin liposomes using magnesium acetate gradient:

[0540] To produce trans-crocetin magnesium liposomes, two different molecular variants can be used, namely: trans-crocetin free acid (TC) and its sodium salt, sodium trans-crocetin (STC).

[0541] Liposomes containing magnesium acetate are prepared by the following procedure. First, the lipid components of the liposome membrane are weighed out and combined in the form of a concentrated solution in ethanol at a temperature of about 65°C. In one example, the lipids used are hydrogenated soybean phosphatidylcholine, cholesterol, and DSPE-PEG-2000 (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy-(polyethylene glycol)-2000]). The molar ratio of HSPC: cholesterol: PEG-DSPE is approximately 3:2:0.15. In another example, the lipids used are HSPC, cholesterol, PEG-DSPE-2000, and 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine (PGPC). The molar ratio of HSPC: cholesterol: PEG-DSPE: PGPC is approximately 2.7:2:0.15:0.3. Next, magnesium acetate is dissolved in an aqueous buffer at a concentration of 125 or 250 mM, with a pH of 7.0. The magnesium acetate solution is heated to 65°C. The ethanol lipid solution is added to the magnesium acetate solution using a pipette. In this step, the solution is stirred thoroughly using a magnetic stirrer. The mixing is performed at an elevated temperature (63°C-72°C) to ensure that the lipids are in a liquid crystal state (as opposed to the gel state they reach at a temperature below the lipid transition temperature Tm=51°C-54°C). As a result, the lipids are hydrated and form multilamellar vesicles (MLVs) containing magnesium acetate in their internal space (internal solution).

[0542] Use filter extrude to reduce the size of the MLV:

[0543] MLVs were fragmented into unilamellar (single bilayer) vesicles of desired size by high pressure extrusion through two passes through a stacked (track-etched polycarbonate) membrane. The stacked membrane had two layers with a pore size of 200 nm and six layers with a pore size of 100 nm. During the extrusion process, the temperature was maintained above Tm. As a result of the extrusion, large and non-uniform MLVs in size and lamellarity were converted into small uniform (100-120 nm) unilamellar vesicles (ULVs) that isolated calcium acetate in their internal space. The hydrodynamic size (diameter) of the vesicles was measured at 25°C in a plastic microcuvette using a Malvern Zetasizer Nano ZS instrument (Southborough, MA) with a backscatter detector (90°). Prior to analysis, the samples were diluted 50 times in the formulation matrix.

[0544] Gradient generation:

[0545] After the production of ULV containing magnesium acetate, the magnesium acetate outside the liposomes is removed using SEC (size exclusion chromatography with a PD-10 column) or TFF (tangential flow diafiltration). A tonicity reagent solution (such as 50% dextrose) is added to the liposomes to balance the osmotic pressure (5% dextrose for 125 mM magnesium acetate liposomes and 10% dextrose for 250 mM magnesium acetate liposomes). The lipid content of the prepared liposome solution is determined by phosphate assay.

[0546] Loading of trans-crocetin into magnesium acetate liposomes:

[0547] Prepare 1 mg / mL trans-crocetin or trans-sodium crocetin solution in 10% dextrose (for 250 mM magnesium acetate liposomes) and adjust the pH to 8-8.5 with sodium hydroxide. Mix the trans-sodium crocetin solution with the magnesium acetate liposome solution at different drug / lipid ratios (100 g / mol, 80 g / mol, 60 g / mol or 40 g / mol). Then stir the mixture thoroughly and heat it to 65°C for 30 minutes, then cool it quickly to room temperature using an ice-water bath. This step can be replaced by stirring the mixture overnight at room temperature.

[0548] Purification of liposomes:

[0549] Trans-crocetin outside the liposome was removed using SEC (PD-10 column) or TFF. In this embodiment, the buffer used in SEC was HBS (HEPES buffered saline, pH 6.5). After purification, filtration sterilization was performed using a 0.2-0.22 micron filter. The hydrodynamic size (diameter) was measured at 25°C in a plastic micro cuvette using a Malvern Zetasizer Nano ZS instrument (Southborough, MA) with a backscatter detector (90°). The sample was diluted before analysis.

[0550] Table 2: Physical properties of representative MTC-loaded nanoparticles

[0551]

[0552] Table 3: Summary of PK results of liposome CTC and MTC

[0553]

[0554] Balb / c mice (3 / group) were treated with a single dose of STC free drug, CTC / MTC-LP (D / L ratio 80, 60, 40) and fluorescent dye-labeled liposomes by slow intravenous push injection to collect serial blood samples at different time points (usually 5 min, 1 hr, 2 hr, 4 hr, 8 hr and 24 hr) over 24 hours.

[0555] 5 μL of each plasma sample was mixed with 395 μL of methanol containing 1% formic acid. The sample mixture was mixed thoroughly by vortexing. The samples were incubated at -20°C for 1 hour and then equilibrated at room temperature for 15 minutes. The samples were vortexed and then centrifuged at 10,000 RPM for 10 minutes at room temperature. 200 μL of supernatant was removed from each sample without disturbing the pellet and analyzed by HPLC. If the amount of plasma allowed, the analysis was repeated.

[0556] The concentration of STC in plasma samples was quantified using a standard curve constructed by analyzing plasma samples containing known amounts of STC.The pharmacokinetic (PK) characteristics were analyzed.

[0557] Table 4: CTC liposome stability

[0558] ...

Claims

1. A pharmaceutical composition comprising an ionizable carotenoid salt having the formula: polyene carotenoid-Q, wherein The polyene carotenoids include (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds, (b) a methyl or lower alkyl (C2-C3) substituent, and (c) 1, 2, 3 or more ionizable groups; and Q is a multivalent counter ion.

2. A pharmaceutical composition comprising an ionizable carotenoid salt having the formula: Q-R1-polyene carotenoid-R2-Q, wherein The polyene carotenoids include (a) 3, 4, 5, 6, 7, 8, 9, 10, 3-5, 6-8, 9-10 or more than 9 conjugated double bonds, (b) a methyl or lower alkyl (C2-C3) substituent, and (c) 1, 2, 3 or more ionizable groups; R1 and R2 are ionizable groups (e.g., the same ionizable group or different ionizable groups; and Q is a multivalent counter ion.

3. The pharmaceutical composition according to claim 1 or 2, wherein the polyene carotenoid comprises 1, 2, 3 or more than 3 anionic ionizable groups.

4. The pharmaceutical composition according to claim 3, wherein the polyene carotenoid comprises at least one anionic ionizable group selected from the group consisting of a carboxyl group, a sulfonate group, a sulfate group, a phosphonate group, a phosphate group and a hydroxamate group.

5. The pharmaceutical composition of claim 1 or 2, wherein the polyene carotenoid comprises 1, 2, 3 or more than 3 cationic ionizable groups (e.g., primary, secondary or tertiary amine groups, quaternary ammonium groups, choline groups, guanidine groups or imidazole groups).

6. A pharmaceutical composition comprising an ionizable carotenoid salt having the formula: Q-trans-crocetin-Q, wherein Q is a multivalent cationic counterion.

7. A pharmaceutical composition comprising an ionizable carotenoid salt having the formula: Q-norbixin-Q, wherein Q is a multivalent cationic counterion.

8. The pharmaceutical composition according to any one of claims 1 to 7, wherein the multivalent counterion (Q) is a multivalent cation (e.g., a divalent cation, such as a divalent metal cation or a divalent organic cation, or a trivalent cation, such as Fe 3+ ).

9. The pharmaceutical composition according to claim 8, wherein the polyvalent cation is at least one selected from Ca 2+ Mg 2+ 、Zn 2+ , Cu 2+ 、Co 2+ and Fe 2+ divalent cations; divalent organic cations, such as protonated diamines; or trivalent cations, such as Fe 3+ .

10. The pharmaceutical composition according to claim 6, comprising magnesium trans-crocetin (MTC) or calcium trans-crocetin (CTC).

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