Patent Application
20250064768
The phenomenon of the increasing number of aging people is arguably the most significant economic, health and social challenge that the world faces today. Approximately 49 million Americans are 65 and older, with projections estimating that the population of older adults will grow to 98 million in 2060. On average, a 65-year old can expect to live another 19 years. The leading causes of death among older adults in the U.S. are chronic diseases-heart disease, cancer, chronic lower respiratory diseases, stroke, Alzheimer's disease, and diabetes, which collectively account for two-thirds of all health care costs in the U.S. and 93% of Medicare spending.
There is a significant unmet need to develop to new therapeutic modalities for slowing or mitigating the effects of aging and for treating age-related diseases or disorders.
The disclosure provides pharmaceutical compositions comprising trans-crocetin and methods of using the compositions to improve health, and to treat aging and chronic disease that comprises administering a plurality of fixed doses of liposomal trans-crocetin to a subject in an amount effective to improve health, treat aging, and/or treat a chronic disease in the subject.
In some embodiments, the disclosure provides a method of improving the general heath in a subject that comprises administering a plurality of fixed doses of liposomal trans-crocetin to a subject in an amount effective to improve the heath of the subject.
In some embodiments, the disclosure provides a method of treating aging in a subject that comprises administering a plurality of fixed doses of liposomal trans-crocetin to the subject in an amount effective to treat aging in the subject.
In some embodiments, the disclosure provides a method of treating a chronic disease that comprises administering a plurality of fixed doses of liposomal trans-crocetin to a subject having or at risk of having the chronic disease in an amount effective to treat the chronic disease.
In a preferred embodiment, the disclosure provides a method of treating aging and/or a chronic disease in a subject that comprises administering a plurality of fixed doses of liposomal trans-crocetin to the subject, wherein the fixed dose is 100 mg to 900 mg, 100 mg to 800 mg, 100 mg to 700 mg 100 mg to 600 mg (e.g., 550 to 600, 560 mg, or 580 mg), 100 mg to 500 mg, 100 mg to 400 mg (e.g., 200 mg to 400 mg, 250 mg to 350 mg, 300 mg to 400 mg, 250 mg, 300 mg, 350 mg, or 380 mg), 100 mg to 300 mg, or 100 mg to 200 mg (e.g., 120 mg to 160 mg or 140 mg), or any range therein between. In some embodiments, a fixed dose of liposomal trans-crocetin is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In a preferred embodiment, the disclosure provides a method of treating aging and/or a chronic disease in a subject that comprises administering a plurality of a plurality of fixed doses of liposomal trans-crocetin to the subject, wherein the fixed dose is 75 mg to 450 mg (e.g., 100 mg to 450 mg, 100 mg to 200 mg, 200 mg to 400 mg, or 140 mg), or any range therein between and wherein each fixed dose is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a preferred embodiment, the disclosure provides a method of treating aging and/or a chronic disease in a subject that comprises administering a plurality of fixed doses of liposomal trans-crocetin to the subject, wherein the fixed dose is 200 mg to 400 mg (e.g., 250 mg to 350 mg, 250 mg, 300 mg, or 380 mg), or any range therein between and wherein each fixed dose is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a preferred embodiment, the disclosure provides a method of treating aging and/or a chronic disease in a subject that comprises administering a plurality of fixed dose of liposomal trans-crocetin to the subject, wherein the fixed dose is 300 mg to 450 mg (e.g., 300 mg to 400 mg, 300 mg, or 380 mg), or any range therein between and wherein each fixed dose is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, a fixed dose of liposomal trans-crocetin is administered four times a month, three times a month, two times a month, or once a month.
In another preferred embodiment, the disclosure provides a method of treating aging and/or a chronic disease in a subject that comprises administering a plurality of fixed doses of liposomal trans-crocetin to the subject, wherein the fixed dose is 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg, or 225 mg to 275 mg (e.g., 250 mg), or any range therein between and wherein each fixed dose is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In one embodiment, a plurality of fixed doses of 120 mg to 160 mg (e.g., 140 mg) liposomal trans-crocetin is administered to the subject and each fixed dose is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In one embodiment, a plurality of fixed doses of 120 mg to 160 mg (e.g., 140 mg) liposomal trans-crocetin is administered to the subject and each fixed dose is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, a fixed dose of liposomal trans-crocetin is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In one embodiment, the disclosure provides a method of treating aging and/or a chronic disease in a subject that comprises administering a plurality of fixed doses of liposomal trans-crocetin to the subject, wherein the fixed dose is 1.5 mg to 250 mg, 1.5 mg to 70 mg, 3 mg to 150 mg, or 5 mg to 240 mg, or any range therein between and wherein each fixed dose is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
The disclosure also provides an article of manufacture comprising at least 1 vial containing a fixed dose of liposomal trans-crocetin, wherein the fixed dose is 1.5 mg to 250 mg, 1.5 mg to 70 mg, 3 mg to 150 mg, or 5 mg to 240 mg, or any range therein between.
The provided trans-crocetin pharmaceutical compositions and dosing regimens that significantly increase the solubility and stability of trans crocetin and thereby significantly expand its therapeutic applications and therapeutic potential. The inventors have surprisingly found that the pharmacokinetic characteristics of trans-crocetin highlight its safety and efficacy across different dosing regimens and show stable levels of AUC and Cmax across a wide range of body weights. The pharmacokinetic profile of trans-crocetin substantiates a fixed dosing strategy for trans-crocetin from a pharmacokinetic perspective.
The provided pharmaceutical compositions and dosing regimens have uses in improving treating aging and/or chronic disease, and in improving health. In some embodiments, the pharmaceutical compositions and dosing regimens have uses in treating conditions associated with aging. In some embodiments, the provided pharmaceutical compositions and dosing regimens have uses in treating disorders and conditions associated a with chronic disease. In some embodiments, the pharmaceutical compositions and dosing regimens have uses in improving the health of a subject. Methods of making, delivering, and using the compositions are also provided.
In some embodiments, the disclosure provides:
In some embodiments, the disclosure provides a method of improving the general heath in a subject that comprises administering a plurality of fixed doses of liposomal trans-crocetin to a subject in an amount effective to improve the heath of the subject.
In some embodiments, the disclosure provides a method of treating aging in a subject that comprises administering a plurality of fixed doses of liposomal trans-crocetin to the subject in an amount effective to treat aging in the subject.
The term “chronic disease(s)” as used herein broadly refers to as conditions that last 1 year or more and require ongoing medical attention or limit activities of daily living or both. Chronic diseases such as heart disease, cancer, and diabetes are the leading causes of death and disability in the United States. Chronic diseases that can be treated according to the provided methods include but are not limited to chronic inflammatory diseases, autoimmune diseases, chronic degenerative diseases such as multiple sclerosis, Parkinson's disease, amyotrophic lateral sclerosis (ALS) and Alzheimer's disease, tumors related to chronic low-grade systemic inflammation (CLGSI) such as breast prostate and colorectal cancers, obesity, diabetes, arterial hypertension, osteoporosis, and chronic infection. In some embodiments, the treated chronic disease is selected from chronic inflammatory disease, rheumatoid arthritis, polymyalgia rheumatica, fibromyalgia, chronic fatigue syndrome and psoriatic arthritis, chronic inflammatory bowel disease, chronic allergic rhinitis, asthma, chronic ulcers, chronic lung disease, chronic heart failure, chronic kidney disease, chronic liver disease, chronic hypoxia, chronic obstructive pulmonary diseases (COPD), emphysema, chronic bronchitis, chronic mountain sickness, cyanotic heart diseases, cystic fibrosis, and obesity, secondary erythrocytosis, chronic graft rejection, chronic organ rejection, chronic wound healing, chronic fatigue syndrome, chronic pain, and diabetes.
In some embodiments, the disclosure provides a method of treating a chronic disease that comprises administering a plurality of fixed doses of liposomal trans-crocetin to a subject having or at risk of having the chronic disease in an amount effective to treat the chronic disease.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is a chronic degenerative disease. In some embodiments, the chronic disease is selected from: Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis (ALS), multiple sclerosis, and dementia.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is a chronic inflammatory disease. In some embodiments, the chronic disease is selected from: rheumatoid arthritis, polymyalgia rheumatica, fibromyalgia, chronic fatigue syndrome and psoriatic arthritis.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is an autoimmune disease. In some embodiments, the chronic disease is selected from: atopic dermatitis, psoriasis and allergic conjunctivitis.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is rheumatoid arthritis, polymyalgia rheumatica, fibromyalgia, chronic fatigue syndrome or psoriatic arthritis.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is an autoimmune disease. In some embodiments, the autoimmune disease is selected from atopic dermatitis, psoriasis and allergic conjunctivitis.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is a degenerative disease. In some embodiments, the degenerative disease is selected from: multiple sclerosis, Parkinson's disease, Amyotrophic lateral sclerosis (ALS) and Alzheimer's disease.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is cancer. In some embodiments, the cancer is selected from: breast, prostate and colorectal cancer.
In some embodiments, the subject treated according to the methods provided herein has, or is diagnosed to one or more risk factors associated with a chronic disease. In some embodiments, the subject has at least one risk factor selected from: smoking, poor nutrition, obesity, diabetes, and excessive alcohol use.
In some embodiments, the disclosure provides methods and dosing regimens in which the provided trans-crocetin compositions are administered in combination therapy with another therapeutic agent.
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.
The Applicants have surprisingly discovered that pharmaceutical compositions such as liposomal compositions containing liposomes comprising multivalent trans-crocetin salts and multivalent counterions substantially improves the pharmacokinetics (e.g., half-life, stability, and bioavailability) and dramatically increases drug exposure via a sustained release of trans-crocetin when compared to for example, trans-crocetin free acids and trans-crocetin salts containing monovalent counterions. The provided pharmaceutical compositions are non-toxic, have surprisingly long extended half lives in vivo, and are able to to reverse/delay symptoms associated with aging and chronic disease. The properties support the extended dosing of the disclosed compositions in treating ageing and chronic disease.
In some embodiments, the disclosure provides a method of improving the general heath in a subject that comprises administering a plurality of fixed doses of liposomal trans-crocetin to a subject in an amount effective to improve the heath of the subject.
In some embodiments, the disclosure provides a method of treating aging in a subject that comprises administering a plurality of fixed doses of liposomal trans-crocetin to the subject in an amount effective to treat aging in the subject.
In some embodiments, the disclosure provides a method of treating a chronic disease that comprises administering a plurality of fixed doses of liposomal trans-crocetin to a subject having or at risk of having the chronic disease in an amount effective to treat the chronic disease.
The term “chronic disease(s)” as used herein broadly refers to as conditions that last 1 year or more and require ongoing medical attention or limit activities of daily living or both. Chronic diseases such as heart disease, cancer, and diabetes are the leading causes of death and disability in the United States. Chronic diseases that can be treated according to the provided methods include but are not limited to chronic inflammatory diseases, autoimmune diseases, chronic degenerative diseases such as multiple sclerosis, Parkinson's disease, amyotrophic lateral sclerosis (ALS) and Alzheimer's disease, tumors related to chronic low-grade systemic inflammation (CLGSI) such as breast prostate and colorectal cancers, obesity, diabetes, arterial hypertension, osteoporosis, and chronic infection. In some embodiments, the treated chronic disease is selected from chronic inflammatory disease, rheumatoid arthritis, polymyalgia rheumatica, fibromyalgia, chronic fatigue syndrome and psoriatic arthritis, chronic inflammatory bowel disease, chronic allergic rhinitis, asthma, chronic ulcers, chronic lung disease, chronic heart failure, chronic kidney disease, chronic liver disease, chronic hypoxia, chronic obstructive pulmonary diseases (COPD), emphysema, chronic bronchitis, chronic mountain sickness, cyanotic heart diseases, cystic fibrosis, and obesity, secondary erythrocytosis, chronic graft rejection, chronic organ rejection, chronic wound healing, chronic fatigue syndrome, chronic pain, and diabetes.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is a chronic degenerative disease. In some embodiments, the chronic disease is selected from: Alzheimer's disease, Parkinson's disease, Amyotrophic lateral sclerosis (ALS), multiple sclerosis, and dementia.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is a chronic inflammatory disease. In some embodiments, the chronic disease is selected from: rheumatoid arthritis, polymyalgia rheumatica, fibromyalgia, chronic fatigue syndrome and psoriatic arthritis.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is an autoimmune disease. In some embodiments, the chronic disease is selected from: atopic dermatitis, psoriasis and allergic conjunctivitis.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is rheumatoid arthritis, polymyalgia rheumatica, fibromyalgia, chronic fatigue syndrome or psoriatic arthritis.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is an autoimmune disease. In some embodiments, the autoimmune disease is selected from atopic dermatitis, psoriasis and allergic conjunctivitis.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is a degenerative disease. In some embodiments, the degenerative disease is selected from: multiple sclerosis, Parkinson's disease, Amyotrophic lateral sclerosis (ALS) and Alzheimer's disease.
In some embodiments, the chronic disease treated or prevented by administering a composition provided herein is cancer. In some embodiments, the cancer is selected from: breast, prostate and colorectal cancer.
In some embodiments, the subject treated according to the methods provided herein has, or is diagnosed to one or more risk factors associated with a chronic disease. In some embodiments, the subject has at least one risk factor selected from: smoking, poor nutrition, obesity, diabetes, and excessive alcohol use.
In some embodiments, the disclosure provides methods and dosing regimens in which the provided trans-crocetin compositions are administered in combination therapy with another therapeutic agent.
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.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the provided compositions, suitable methods and materials are described below. Each publication, patent application, patent, and other reference mentioned herein is herein incorporated by reference in its entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be limiting.
Other features and advantages of the disclosed compositions and methods will be apparent from the following disclosure, drawings, and claims.
It is understood that wherever embodiments, are described herein with the language “comprising” otherwise analogous embodiments, described in terms of “containing” “consisting of” and/or “consisting essentially of” are also provided. However, when used in the claims as transitional phrases, each should be interpreted separately and in the appropriate legal and factual context (e.g., in claims, the transitional phrase “comprising” is considered more of an open-ended phrase while “consisting of” is more exclusive and “consisting essentially of” achieves a middle ground).
As used herein, the singular form “a”, “an”, and “the”, include plural forms unless it is expressly stated or is unambiguously clear from the context that such is not intended. The singular form “a”, “an”, and “the” also includes the statistical mean composition, characteristics, or size of the particles in a population of particles (e.g., mean liposome diameter, mean liposome zeta potential, mean number of targeting moieties on liposomes in a liposomal solution, mean number of encapsulated trans-crocetin molecules). The mean particle size and zeta potential of liposomes in a pharmaceutical composition can routinely be measured using methods known in the art, such as dynamic light scattering. The mean amount of a therapeutic agent in a nanoparticle composition may routinely be measured for example, using absorption spectroscopy (e.g., ultraviolet-visible spectroscopy).
As used herein, the terms “approximately” and “about,” as applied to one or more values of interest, refer to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” refers 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 in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value). For example, when used in the context of an amount of a given compound in a lipid component of a nanoparticle composition, “about” may mean+/−10% of the recited value. For instance, a nanoparticle composition including a lipid component having about 40% of a given compound may include 30-50% of the compound.
The term “and/or” as used in a phrase such as “A and/or B” herein is intended to include both A and B; A or B; A (alone); and B (alone). Likewise, the term “and/or” as used in a phrase such as “A. B, and/or C” is intended to encompass 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).
Where embodiments, of the disclosure are described in terms of a Markush group or other grouping of alternatives, the disclosed composition or method encompasses not only the entire group listed as a whole, but also each member of the group individually and all possible subgroups of the main group, and also the main group absent one or more of the group members. The disclosed compositions and methods also envisage the explicit exclusion of one or more of any of the group members in the disclosed compositions or methods.
The term “complexed” as used herein relates to the non-covalent interaction of a biomacromolecule agent such as trans-crocetin with a polymer such as a cyclodextrin or phospholipid.
As used herein, the term “cyclodextrin” includes any of the known cyclodextrins and cyclodextrin derivatives, such as unsubstituted cyclodextrins containing from six to twelve glucose units, especially, alpha-cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin and/or their derivatives and/or mixtures thereof. Unless otherwise specified herein, “cyclodextrin” refers generally to a parent or derivatized cyclic oligosaccharide containing a variable number of (α-1,4)-linked D-glucopyranoside units that is able to form a complex with trans-crocetin.
Each cyclodextrin glucopyranoside subunit has secondary hydroxyl groups at the 2 and 3 positions and a primary hydroxyl group at the 6-position. The terms “parent,” “underivatized,” or “inert,” cyclodextrin refer to a cyclodextrin containing D-glucopyranoside units having the basic formula C6H12O6 and a glucose structure without any additional chemical substitutions (e.g., α-cyclodextrin consisting of 6 D-glucopyranoside units, a β-cyclodextrin consisting of 7 D-glucopyranoside units, and a 7-cyclodextrin consisting of 8 D-glucopyranoside units). The physical and chemical properties of a parent cyclodextrin can be modified by derivatizing the hydroxyl groups with other functional groups. In particular embodiments, the administered compositions comprise trans-crocetin complexed with 7-cyclodextrin.
The term “conjugated” as used herein indicates a covalent bond association between a biomacromolecule such as trans-crocetin and a polymer such as polyethylene glycol or an antibody or other polypeptide. In some embodiments, the trans-crocetin conjugate displays increased half-life and/or targeting specificity compared to unconjugated trans-crocetin.
The term “encapsulated” as used herein refers to the location of a biomacromolecule agent (e.g., trans-crocetin) that is enclosed or completely contained within the inside of a polymer such as a liposome.
The term “liposome” refers to a closed vesicle having an internal phase (i.e., interior space (internal solution)) enclosed by lipid bilayer. A liposome can be a small single-membrane liposome such as a small unilamellar vesicle (SUV), large single-membrane liposome such as a large unilamellar vesicle (LUV), a still larger single-membrane liposome such as a giant unilamellar vesicle (GUV), a multilayer liposome having multiple concentric membranes (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10), such as a multilamellar vesicle (MLV), or a liposome having multiple membranes that are irregular and not concentric such as a multivesicular vesicle (MVV). Liposomes and liposome formulations are well known in the art. Lipids which are capable of forming liposomes include all substances having fatty or fat-like properties. Lipids which can make up the lipids in the liposomes include without limitation, glycerides, glycerophospholipids, glycerophosphinolipids, glycerophos-phonolipids, sulfo-lipids, sphingolipids, phospholipids, isoprenolides, steroids, stearines, sterols, archeolipids, synthetic cationic lipids and carbohydrate containing lipids.
A “liposome composition” is a prepared composition comprising a liposome and the contents within the liposome, particularly including the lipids which form the liposome bilayer(s), compounds other than the lipids within the bi-layer(s) of the liposome, compounds within and associated with the aqueous interior(s) of the liposome, and compounds bound to or associated with the outer layer of the liposome. Thus, in addition to the lipids of the liposome, a liposome composition described herein suitably may include, but is not limited to, therapeutic agents, immunostimulating agents, vaccine antigens and adjuvants, excipients, carriers and buffering agents. In a preferred embodiment, such compounds are complementary to and/or are not significantly detrimental to the stability or AGP-incorporation efficiency of the liposome composition.
The terms liposome “internal phase”, “interior space”, and “internal core” are used interchangeably to refer to an aqueous region enclosed within (i.e., encapsulated by) the lipid bilayer of the liposome. The solution of the liposomal internal phase is referred to as the “internal solution.” By contrast, the term “liposome external phase” refers to the region not enclosed by the lipid bilayer of the liposome, such as the region apart from the internal phase and the lipid bilayer in the case where the liposome is dispersed in liquid.
The term “counterion” refers to an anionic or cationic counterion.
A “cationic counterion” is a positively charged atom or group associated with an anionic atom or group in order to maintain electronic neutrality. Exemplary cationic counterions include inorganic cations (e.g., metal cations (e.g., alkali metal cations, alkali 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 associated with a cationic atom or group in order to maintain electronic neutrality. Exemplary anionic counterions include halide anions (e.g., F—, Cl—, Br—, and I—), NO3−, ClO4−OH—, H2PO4−2, HSO4−, sulfonate anions (e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphor sulfonate, naphthalene-2-sulfonate, naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonic acid-2-sulfonate, and the like), and carboxylate anions (e.g., acetate, ethanoate, propanoate, benzoate, glycerate, lactate, tartrate, and glycolate). A counterion may be monovalent or multivalent (e.g., divalent, trivalent, tetravalent, etc.).
The term “ionizable” refers to a compound containing at least one functional group that (a) bears a positive or negative charge (i.e., is “ionized”) and is therefore associated with a counterion of opposite charge, or (b) is electronically neutral but ionized at a higher or lower pH. Thus, ionizable compounds include quaternary ammonium salts as well as uncharged amines, and carboxylate moieties as well as uncharged carboxyl groups.
The term “naturally occurring” refers to a compound or composition that occurs in nature, regardless of whether the compound or composition has been isolated from a natural source or chemically synthesized. Examples of naturally occurring carotenoid mono- and di-carboxylic acids include crocetin, norbixin, azafrin and neurosporaxanthin.
As used herein an “effective amount” refers to a dosage of an agent sufficient to provide a medically desirable result. The effective amount will vary with the desired outcome, the particular disease or condition being treated or prevented, the age and physical condition of the subject being treated, the severity of the condition, the duration of the treatment, the nature of the concurrent or combination therapy (if any), the specific route of administration and like factors within the knowledge and expertise of the health practitioner. An “effective amount” can be determined empirically and in a routine manner, in relation to the stated purpose.
In the case of a pulmonary disorder such as ARDS, COPD, sepsis or pulmonary inflammation, an effective amount of an agent may for example, stabilize or improve lung function, such as demonstrated by physical examination and respiratory rate normalization, improving pAO2/FiO2 ratio (P/F ratio, e.g., show an increase of at least 15%, 20%, 25% PaO2/FiO2 ratio increase or a PaO2/FiO2 ratio increase above 200 mm Hg within 24 hours after administration), normalization of pCO2, preventing need for intubation and mechanical ventilation, (for those mechanically ventilated) decreased number of ventilator days, decreased hospital length of stay, decreased intensive care unit length of stay, or a combination thereof.
In the case of cancer, the effective amount of an agent may for example, reduce the number of cancer cells; reduce the tumor size; inhibit (i.e., slow to some extent and preferably stop) cancer cell infiltration into peripheral organs; inhibit (i.e., slow to some extent and preferably stop) tumor metastasis; inhibit, to some extent, tumor growth; and/or relieve to some extent one or more of the symptoms associated with the disorder. To the extent the drug may prevent growth and/or kill existing cancer cells, it may be cytostatic and/or cytotoxic. For cancer therapy, efficacy in vivo can, for example, be measured by assessing the duration of survival, duration of progression free survival (PFS), the response rates (RR), duration of response, and/or quality of life.
As used herein, the phrase “a subject in need thereof” means a subject (e.g., human or non-human mammal that exhibits one or more symptoms or indications of, or has been identified as having aging and/or a chronic disease and thereby having a need for the particular method or treatment. In some embodiments, the diagnosis can be by any means of diagnosis. In any of the methods and treatment regimens described herein, the subject can be in need thereof. In some embodiments, the term “a subject in need thereof” includes, e.g., patients who, prior to treatment, exhibit (or have exhibited) one or more symptoms of an age-related disease or condition, and/or who exhibit senescence biomarkers and/or SASP. In some embodiments, the term “a subject in need thereof” includes, e.g., patients who, prior to treatment, exhibit (or have exhibited) one or more symptoms of a chronic disease or condition, and/or who exhibit a chronic disease biomarker.
The terms “hyperproliferative disorder”, “proliferative disease”, and “proliferative disorder”, are used interchangeably herein to pertain to an unwanted or uncontrolled cellular proliferation of excessive or abnormal cells which is undesired, such as, neoplastic or hyperplastic growth, whether in vitro or in vivo. In some embodiments, the proliferative disease is cancer or tumor disease (including benign or cancerous) and/or any metastases, wherever the cancer, tumor and/or the 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 hyperplasias, fibrosis (especially pulmonary, but also other types of fibrosis, such as renal fibrosis), angiogenesis, psoriasis, atherosclerosis and smooth muscle proliferation in the blood vessels, such as stenosis or restenosis following angioplasty.
“Cancer,” “tumor,” or “malignancy” are used as synonymous terms and refer to any of a number of disorders that are characterized by uncontrolled, abnormal proliferation of cells, the ability of affected cells to spread locally or through the bloodstream and lymphatic system to other parts of the body (metastasize) as well as 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 pre-cancerous and cancerous cells and tissues. A “cancerous tumor,” or “malignant cell” is understood as a cell having specific structural properties, lacking differentiation and being capable of invasion and metastasis. A cancer that can be treated using a liposomal trans-crocetin composition and/or dosing regimen provided herein includes without limitation, a non-hematologic malignancy including such as 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, biliary duct cancer, gallbladder cancer, bladder cancer, sarcoma (e.g., osteosarcoma), brain cancer, central nervous system cancer, and melanoma; and a hematologic malignancy such as for example, a leukemia, a lymphoma and other B cell malignancies, myeloma and other plasma cell dysplasias or dyscrasias. Other types of cancer and tumors that may be treated using a trans-crocetin composition are described herein or otherwise known in the art. The terms “cancer,” “cancerous,” “cell proliferative disorder,” “proliferative disorder,” and “tumor” are not mutually exclusive as referred to herein.
“Ischemia” relates to a restriction in blood supply to tissues or organs (tissue hypoperfusion) causing a shortage of oxygen needed for cellular metabolism. The term “ischemia injury”, as used herein, relates to the damage due to a shortage of oxygen needed for cellular metabolism and to conditions associated with ischemia, including, but not limited to ischemic stroke, peripheral vascular disease, cerebral vascular disease, kidney disease and ischemia associated renal pathologies, reperfusion, reperfusion injury, ischemia associated with wounds, tissue hypoperfusion, ischemic-reperfusion injury, transient cerebral ischemia, cerebral ischemia-reperfusion, ischemic stroke, hemorrhagic stroke, traumatic brain injury, gastrointestinal ischemia, pulmonary embolism, acute respiratory failure, neonatal respiratory distress syndrome, an obstetric emergency to reduce perinatal comorbidity (such as, pre/eclampsia and conditions that lead to cerebral palsy), myocardial infarction, acute limb or mesenteric ischemia, coronary artery disease, cardiac cirrhosis, chronic congestive heart failure, atherosclerotic stenosis, anemia, thrombosis, embolism, or migraine (e.g., a chronic migraine or severe migraine disorder).
Organ “impairment” or dysfunction” refers to a condition wherein a particular organ does not perform its expected function. An organ dysfunction develops into organ failure if the normal homeostasis cannot be maintained without external clinical intervention. Methods to determine organ dysfunction are known in the art and include without limitation, monitorization and scores including sequential organ failure assessment (SOFA) score, multiple organ dysfunction (MOD) score and logistic organ dysfunction (LOD) score.
Terms such as “treating,” or “treatment,” or “to treat” refer to both (a) therapeutic measures that cure, slow down, attenuate, lessen symptoms of, and/or halt progression of a diagnosed pathologic disorder or condition and (b) prophylactic or preventative measures that prevent and/or slow the development of a targeted disorder or condition. The desirable effects of the treatment include, but are not limited to, the prevention of the development or recurrence of aging and/or a chronic disease, the alleviation of symptoms associated with the disorder or conditions, the attenuation of any direct or indirect pathological influence of the disorder or condition, the prevention of metastasis, reduction in the rate of progression of the disorder or condition, recovery from or alleviation of aging and/or a chronic disease, and/or ameliorated or improved prognosis. Thus, subjects in need of treatment include for example, those already with a chronic disease, those at risk of having a chronic disease, and those in whom the chronic disease is to be prevented. Subjects are identified as “having or at risk of having” for example, a chronic disease using well-known medical and diagnostic techniques. In certain embodiments, a subject is successfully “treated” according to the methods provided herein if the subject shows, e.g., total, partial, or transient amelioration or elimination of a symptom associated with the disease or condition (e.g., aging and/or a chronic disease). Treatment can be with a provided pharmaceutical composition disclosed herein (e.g., a liposomal trans-crocetinate) alone, or in combination with an additional therapeutic agent.
The terms “subject” and “patient,” and “animal” are used interchangeably and refer to mammals such as human patients and non-human primates, as well as experimental animals such as rabbits, rats, and mice, and other animals. Animals include all vertebrates, e.g., mammals and non-mammals, such as chickens, amphibians, and reptiles. “Mammal” as used herein refers to any member of the class Mammalia, including, without limitation, humans and nonhuman primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, sheep, pigs, goats and horses; domestic mammals such as dogs and cats; laboratory animals including rodents such as mice, rats and guinea pigs, and other members of the class Mammalia known in the art. In a particular embodiment, the patient is a human.
The term “elderly” refers to an aged subject, who has passed middle age. In one embodiment, an elderly mammalian subject is a subject that has survived more than two-thirds of the normal lifespan for that mammalian species. In a further embodiment, for humans, an aged or elderly subject is more than 65 years of age, such as a subject of more than 70, more than 75, more than 80 years of age. In yet another embodiment, for mice, an elderly mouse is from about 14 to about 18 months of age.
A “fixed” or “flat” dose or “fixed dosing” or “flat dosing” of a therapeutic agent such as trans-crocetin and/or another therapeutic agent refers to a dose that is administered to a subject such as a human patient without regard for the weight (WT) or body surface area (BSA) of subject patient. The fixed or flat dose is therefore not provided as a mg/kg dose or a mg/m dose, but rather as an absolute amount of the liposomal trans-crocetin and/or another therapeutic agent.
In some embodiments, the fixed dosing regimen can be stratified into two or more fixed dose amounts based on a patient's body weight, body mass, age, gender, frame size, height, general health, or any combinations thereof, or independently of age, height, gender, frame size, general health. For example, the fixed dose can be stratified into multiple fixed dose amounts (e.g., three) suitable for subjects who fall within the weight categories, e.g., those with low, normal, or high body weight. In one embodiment, the fixed dose amounts are stratified based on the weight, categories of low body weight (e.g., 100 pounds or less, normal body weight (e.g., between 100 and 190 pounds, and high body weight (e.g., more than 190 pounds. In another embodiments, the fixed dose amounts are stratified based on the age and sex categories of child under 12, male from 10 to 15 and female over 12, and male over 15 years of age. A person of ordinary skill in the art can assess the factors related to body weight and can determine the specific body weight category for a subject.
A “loading dose” refers to an amount of a therapeutic agent such as trans-crocetin (e.g., a liposomal trans-crocetin, free trans-crocetin, conjugated/complexed trans-crocetin, or a combination thereof), administered to a subject during the initial stages of the treatment. The loading dose(s) of trans-crocetin can be fixed or not fixed. The purpose of a loading dose is to more rapidly achieve therapeutic levels of therapeutic agent in the subject than that which would have been reached with maintenance dosing only. In addition, the loading dose can also achieve sufficient levels of therapeutic agent (e.g., trans-crocetin) to enable therapeutic levels to be maintained once the switch to maintenance dosing is made. Furthermore, the loading dose allows a relatively constant therapeutic level of therapeutic agent to be achieved in which therapeutic agent is in a steady-state. The steady state of therapeutic agent may be regarded as a state in which the overall intake of therapeutic agent is in dynamic equilibrium with its elimination. Once therapeutic levels of therapeutic agent are reached, the loading doses may be followed by a plurality of fixed maintenance doses. Increasing the loading dose concentration may allow the time intervals between the loading doses to be extended.
A “maintenance dose” refers to an amount of a therapeutic agent such as liposomal trans-crocetin administered to a subject over a treatment period in order to maintain therapeutic levels of therapeutic agent in the subject. Such therapeutic levels of therapeutic agent are achieved and maintained more rapidly by providing the subject with one or more loading doses as described herein prior to providing the maintenance dose(s). Usually, the maintenance doses are administered at spaced treatment intervals. In some embodiments, maintenance doses of trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, maintenance doses of trans-crocetin is administered to a subject four times a month, three times a month, two times a month, or once a month.
The term “pharmaceutically acceptable carrier” refers to an ingredient in a pharmaceutical formulation, other than an active ingredient, which is nontoxic to a subject. A pharmaceutically acceptable carrier includes, but is not limited to, a buffer, carrier, excipient, stabilizer, diluent, or preservative. Pharmaceutically acceptable carriers can include for example, one or more compatible solid or liquid filler, diluents or encapsulating substances which are suitable for administration to a human or other subject.
The term “therapeutic agent” is used herein in its broadest sense to include any agent capable of providing a desired or beneficial effect on a subject. Thus, the term includes both prophylactic and therapeutic agents, as well as any other category of agent having such desired effects, therapeutic agent or therapeutic agents used in combination therapy with trans-crocetin according to the disclosed compositions and methods can include any agent directed to treat a condition in a subject.
The term “kit” refers to a set of one or more components necessary for employing the methods and compositions provided herein. Kit components can include, but are not limited to, trans-crocetin compositions including liposomal trans-crocetin, cyclodextrin-trans-crocetin formulations, and free trans-crocetin formulations disclosed herein, reagents, buffers, containers and/or equipment.
The term “radiosensitizing agent” means a compound that makes tumor cells more sensitive to radiation therapy. Examples of radiosensitizing agents include misonidazole, metronidazole, tirapazamine, and trans-crocetin.
In an additional embodiment, the disclosure provides an article of manufacture comprising materials useful for the treatment of aging and/or a chronic disease described herein (e.g., aging and/or a chronic disease described herein). The article of manufacture comprises a vial containing a fixed dose of liposomal trans-crocetin and optionally a package insert. The vial may be formed from a variety of materials, such as glass or plastic, and may be sealed by a syringe with a stopper that can be punctured. In further embodiments, the article of manufacture may contain other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, and the like. In one embodiment, the article of manufacture comprises a vial containing a fixed dose of liposomal trans-crocetin (e.g., liposomal trans-crocetin or gamma cyclodextrin complexed trans-crocetin), wherein the fixed dose is for example, approximately 140 mg, 420 mg, 500 mg, 525 mg, 840 mg, or 1050 mg of trans-crocetin. In one embodiment, the article of manufacture comprises one or more vials containing a fixed dose of approximately 140 mg liposomal trans-crocetin. In one embodiment, the article of manufacture comprises one or more vials containing a fixed dose of approximately 250 mg liposomal trans-crocetin. In one embodiment, the article of manufacture comprises one or more vials containing a fixed dose of approximately 300 mg liposomal trans-crocetin. In one embodiment, the article of manufacture comprises one or more vials containing a fixed dose of approximately 140 mg liposomal trans-crocetin and/or one or more vials containing a fixed dose of approximately 250 mg liposomal trans-crocetin. In one embodiment, the article of manufacture comprises one or more vials containing a fixed dose of approximately 140 mg liposomal trans-crocetin and/or one or more vials containing a fixed dose of approximately 300 mg liposomal trans-crocetin.
In a further particular embodiment, the article of manufacture contains one or more vials containing a fixed dose of liposomal trans-crocetin of approximately 140 mg and/or one or more vials containing a fixed dose of liposomal trans-crocetin at approximately 250 mg.
In some embodiments, the article of manufacture further comprises a package insert. The package inserts may provide instructions for administering a plurality of fixed doses of liposomal trans-crocetin pharmaceutical composition and/or for administering compositions according to the dosing regimens provided herein, of the invention. The package inserts may also provide drug preparation and/or dosing instructions for treating disorders and conditions such as aging and/or a chronic disease described herein.
The term “pharmaceutical composition” as used herein usually refers to a drug for the treatment or prevention of a disease or condition, or for examination or diagnosis. The provided pharmaceutical compositions can be prepared in a variety of ways using commercially available starting materials, compounds known in the literature, or from readily prepared intermediates, by employing standard synthetic methods and procedures either known to those skilled in the art, or which will be apparent to the skilled artisan in light of the teachings herein. Standard synthetic methods and procedures for the preparation of organic molecules and 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 several sources, classic texts such as Smith et al., March's Advanced Organic Chemistry. Reactions, Mechanisms, and Structure, 5thedition, John Wiley & Sons: New York, 2001; Greene, T. W., Wuts, P. G. M., 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), incorporated by reference herein, are useful and recognized reference textbooks of organic synthesis known to those in the art. The following descriptions of synthetic methods are designed to illustrate, but not to limit, general procedures for the preparation of compounds of the present disclosure.
In some embodiments, the pharmaceutical composition comprises a trans-crocetin having the formula: Q-trans-crocetin-Q
wherein, Q is a multivalent cation counterion. In some embodiments, the pharmaceutical composition administered according to the provided methods is a fixed dose of liposomal trans-crocetin. In some embodiments, the administered liposomal trans-crocetin composition does not contain a fixed dose of liposomal trans-crocetin.
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 cation counterion. In further embodiments, Q is a divalent metal cation. In some embodiments, Q is at least one member selected from Ca2+, Mg2+, Zn2+, Cu2+, Co2+, and Fe2+. In further embodiments, Q is Ca2+ or Mg2+. In some embodiments, Q is Ca2+. In some embodiments, Q is Mg2+. In other embodiments, Q is a trivalent cation counterion such as Fe3+. In some embodiments, Q is a multivalent organic cation. In further embodiments, Q is a divalent organic cation such as a protonated diamine. Liposomes comprising the trans-crocetin compositions and pharmaceutical compositions (e.g., liposome compositions) comprising the liposomes are also provided herein. In some embodiments, the pharmaceutical composition administered according to the provided methods is a fixed dose of liposomal trans-crocetin. In some embodiments, the administered liposomal trans-crocetin composition does not contain a fixed dose of liposomal trans-crocetin.
In some embodiments, the disclosure provides a pharmaceutical composition comprising calcium trans-crocetin (CTC). The CTC can exist in linear and/or cyclic form (shown below)
Liposomes comprising the CTC compositions and pharmaceutical compositions (e.g., liposome compositions) comprising the liposomes are also provided herein. In some embodiments, the pharmaceutical composition administered according to the provided methods is a fixed dose of liposomal CTC. In some embodiments, the administered liposomal trans-crocetin composition does not contain a fixed dose of liposomal CTC.
In some embodiments, the disclosure provides a pharmaceutical composition comprising magnesium trans-crocetin (MTC). The MTC can exist in linear and/or cyclic form (shown below).
Liposomes comprising the MTC compositions and pharmaceutical compositions (e.g., liposome compositions) comprising the liposomes are also provided herein. In some embodiments, the pharmaceutical composition administered according to the provided methods is a fixed dose of liposomal MTC. In some embodiments, the administered liposomal trans-crocetin composition does not contain a fixed dose of liposomal MTC.
The lipids and other components of the liposomes contained in the liposome compositions can be any lipid, lipid combination and ratio, or combination of lipids and other liposome components and their respective ratios known in the art. However, it will be understood by one skilled in the art that liposomal encapsulation of any particular drug, such as, and without limitation, the trans-crocetin compositions discussed herein, may involve substantial routine experimentation to achieve a useful and functional liposomal formulation. In general, the provided liposomes may have any liposome structure, e.g., structures having an inner space sequestered from the outer medium by one or more lipid bilayers, or any microcapsule that has a semi-permeable membrane with a lipophilic central part where the membrane sequesters an interior. The lipid bilayer can be any arrangement of amphiphilic molecules characterized by a hydrophilic part (hydrophilic moiety) and a hydrophobic part (hydrophobic moiety). Usually, amphiphilic molecules in a bilayer are arranged into two dimensional sheets in which hydrophobic moieties are oriented inward the sheet while hydrophilic moieties are oriented outward. Amphiphilic molecules forming the provided liposomes can be any known or later discovered amphiphilic molecules, e.g., lipids of synthetic or natural origin or biocompatible lipids. The liposomes can also be formed by amphiphilic polymers and surfactants, e.g., polymerosomes and niosomes. For the purpose of this disclosure, without limitation, these liposome-forming materials also are referred to as “lipids”.
The liposome composition formulations provided herein can be in liquid or dry form such as a dry powder or dry cake. The dry powder or dry cake may have undergone primary drying under, for example, lyophilization conditions or optionally, the dry cake or dry powder may have undergone both primary drying only or both primary drying and secondary drying. In the dry form, the powder or cake may, for example, have between 1% to 6% moisture, for example, such as between 2% to 5% moisture or between 2% to 4% moisture. One example method of drying is lyophilization (also called freeze-drying, or cryodessication). Any of the compositions and methods of the disclosure may include liposomes, lyophilized liposomes or liposomes reconstituted from lyophilized liposomes. In some embodiments, the compositions and methods include one or more lyoprotectants or cryoprotectants. These protectants are typically polyhydroxy compounds such as sugars (mono-, di-, and polysaccharides), polyalcohols, and their derivatives, glycerol, or polyethyleneglycol, trehalose, maltose, sucrose, glucose, lactose, dextran, glycerol, or aminoglycosides. In further embodiments, the lyoprotectants or cryoprotectants comprise up to 10% or up to 20% of a solution outside the liposome, inside the liposome, or both outside and inside the liposome.
The properties of liposomes are influenced by the nature of lipids used to make the liposomes. A wide variety of lipids have been used to make liposomes. These include cationic, anionic and neutral lipids. In some embodiments, the liposomes comprising the trans-crocetin compositions (e.g., CTC and MTC) are anionic or neutral. In other embodiments, the provided liposomes are cationic. The determination of the charge (e.g., anionic, neutral or cationic) can routinely be determined by measuring the zeta potential of the liposome. The zeta potential of the liposome can be positive, zero or negative. In some embodiments, the zeta potential of the liposome is −150 to 150 mV, or −50 to 50 mV, or any range therein between. 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 therein between. In other embodiments, the zeta potential of the liposome is more than zero. In some embodiments, the liposome has a zeta potential that is 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 therein between.
Depending on the desired application, the particle size (diameter) of the liposome can be regulated. For example, when it is intended to deliver the liposome to cancerous tissue or inflamed tissue by the Enhanced Permeability and Retention (EPR) effect as an injection product or the like, it is preferable that liposome diameter is 20-500 nm, 30-175 nm, or 50-150 nm, or any range therein between. In the case where the intention is to transmit liposome to macrophage, it is preferable that liposome diameter is 30 to 1000 nm, or 80 to 400 nm, or any range therein between. In the case where liposome composition is to be used as an oral preparation or transdermal preparation, the particle size of liposome can be set at several microns. It should be noted that in normal tissue, vascular walls serve as barriers (because the vascular walls are densely constituted by vascular endothelial cells), and microparticles such as supermolecules and liposome of specified size cannot be distributed within the tissue. However, in diseased tissue, vascular walls are loose (because interstices exist between vascular endothelial cells), increasing vascular permeability, and supermolecules and microparticles can be distributed to extravascular tissue (enhanced permeability). Moreover, the lymphatic system is well developed in normal tissue, but it is known that the lymphatic system is not developed in diseased tissue, and that supermolecules or microparticles, once incorporated, are not recycled through the general system, and are retained in the diseased tissue (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)). Thus, it is possible to control liposome pharmacokinetics by adjusting liposome particle size (diameter).
In some embodiments, cationic lipids are used to make cationic liposomes which are commonly used as gene transfection agents. The positive charge on cationic liposomes enables interaction with the negative charge on cell surfaces. Following binding of the cationic liposomes to the cell, the liposome is transported inside the cell through endocytosis.
In some preferred embodiments, a neutral to anionic liposome is used. In a preferred embodiment, an anionic liposome is used. Using a mixture of, for example, 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 a tumor targeting antibody such as, for example, a folate receptor antibody, including, for example, folate receptor alpha antibody, folate receptor beta antibody and/or folate receptor delta antibody.
As an example, at least one (or some) of the lipids is/are amphipathic lipids, defined as having a hydrophilic and a hydrophobic portion (typically a hydrophilic head and a hydrophobic tail). The hydrophobic portion typically orients into a hydrophobic phase (e.g., within the bilayer), while the hydrophilic portion typically orients toward the aqueous phase (e.g., outside the bilayer). The hydrophilic portion can comprise polar or charged groups such as carbohydrates, phosphate, carboxylic, sulfate, amino, sulfhydryl, nitro, hydroxy and other like groups. The hydrophobic portion can comprise apolar groups that include without limitation long chain saturated and unsaturated aliphatic hydrocarbon groups and groups substituted by one or more aromatic, cyclo-aliphatic or heterocyclic group(s). Examples of amphipathic compounds include, but are not limited to, phospholipids, aminolipids and sphingolipids.
Typically, for example, the lipids are phospholipids. Phospholipids include without limitation phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, and the like. It is to be understood that other lipid membrane components, such as cholesterol, sphingomyelin, and cardiolipin, can be used.
The lipids comprising the liposomes provided herein can be anionic and neutral (including zwitterionic and polar) lipids including anionic and neutral phospholipids. Neutral lipids exist in an uncharged or neutral zwitterionic form at a selected pH. At physiological pH, such lipids include, for example, dioleoylphosphatidylglycerol (DOPG), diacylphosphatidylcholine, diacylphos-phatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebrosides and diacylglycerols. Examples of zwitterionic lipids include without limitation dioleoylphosphatidylcholine (DOPC), dimyristoylphos-phatidylcholine (DMPC), and dioleoylphosphatidylserine (DOPS). Anionic lipids are negatively charged at physiological pH. These lipids include without limitation phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacyl-phosphatidic acid, N-dodecanoyl phosphatidylethanolamines, N-succinyl phosphatidylethanolamines, N-glutarylphosphatidylethanolamines, lysylphosphatidyl-glycerols, palmitoyloleyo-lphosphatidylglycerol (POPG), and other anionic modifying groups joined to neutral lipids.
Collectively, anionic and neutral lipids are referred to herein as non-cationic lipids. Such lipids may contain phosphorus but they are not so limited. Examples of non-cationic lipids include lecithin, lysolecithin, phosphatidylethanolamine, lysophosphatidyl-ethanolamine, dioleoylphosphatidylethanolamine (DOPE), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoylphosphatidy 1-ethanolamine (DSPE), palmitoyloleoyl-phosphatidylethanolamine (POPE) palmitoyl-oleoylphosphatidylcholine (POPC), egg phosphatidylcholine (EPC), distearoylphospha-tidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidyl-choline (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoyl-phosphatidyl-glycerol (DPPG), palmitoyloleyol-phosphatidylglycerol (POPG), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans-PE, palmitoyloleoylphosphatidylethanolamine (POPE), 1-stearoyl-2-oleoylphosphatidylethanolamine (SOPE), phosphatidylserine, phosphatidyl-inositol, sphingomyelin, cephalin, cardiolipin, phosphatidic acid, cerebrosides, dicetylphosphate, and cholesterol.
The liposomes may be assembled using any liposomal assembly method using liposomal components (also referred to as liposome components) known in the art. Liposomal components include, for example, lipids such as DSPE, HSPC, cholesterol and derivatives of these components. Other suitable lipids are commercially available for example, by Avanti Polar Lipids, Inc. (Alabaster, Alabama, USA). A partial listing of available negatively or neutrally charged lipids suitable for making 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.
In some embodiments, the provided compositions are formulated in a liposome comprising a cationic lipid. In one embodiment, the cationic lipid is selected from, but not limited to, a cationic lipid described in Intl. Publ. Nos. WO2012/040184, WO2011/153120, WO2011/149733, WO2011/090965, WO2011/043913, WO2011/022460, WO2012/061259, WO2012/054365, WO2012/044638, WO2010/080724, WO2010/21865 and WO2008/103276, U.S. Pat. Nos. 7,893,302, 7,404,969 and 8,283,333, and U.S. Appl. Publ. Nos. US20100036115 and US20120202871; each of which is herein incorporated by reference in its entirety. In another embodiment, the cationic lipid may be selected from, but not limited to, formula A described in Intl. Appl. Publ. Nos. WO2012/040184, WO2011/153120, WO201/1149733, WO2011/090965, WO2011/043913, WO2011/022460, WO2012/061259, WO2012/054365 and WO2012/044638; each of which is herein incorporated by reference in its entirety. In yet another embodiment, the cationic lipid may be selected from, but 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 US Publ. No. US20100036115; each of which is herein incorporated by reference in its entirety. As a non-limiting example, the cationic lipid may be selected from (20Z,23Z)-N,N-dimethylnonacosa-20,23-dien-10-amine, (17Z,20Z)-N,N-dimemyl-hexacosa-17,20-dien-9-amine, (1Z,19Z)-N5N-dimethyl-pentacosa-16, 19-dien-8-amine, (13Z,16Z)-N,N-dimethyldocosa-13,16-dien-5-amine, (12Z,15Z)-N,N-dimethylhenicosa-12,15-dien-4-amine, (14Z,17Z)-N,N-dimethyltricosa-14,17-dien-6-amine, (15Z,18Z)-N,N-dimethyltetracosa-15,18-dien-7-amine, (18Z,21Z)-N,N-dimethylheptacosa-18,21-dien-10-amine, (15Z,18Z)-N,N-dimethyltetracosa-15,18-dien-5-amine, (14Z,17Z)-N,N-dimethyl-tricosa-14,17-dien-4-amine, (19Z,22Z)-N,N-dimethyloctacosa-19,22-dien-9-amine, (18Z,21 Z)-N,N-dimethylheptacosa-18,21-dien-8-amine, (17Z,20Z)-N,N-dimethylhexacosa-17,20-dien-7-amine, (16Z,19Z)-N,N-dimethylpenta-cosa-16,19-dien-6-amine, (22Z,25Z)-N,N-dimethylhentriaconta-22,25-dien-10-amine, (21 Z,24Z)-N,N-dimethyl-triaconta-21,24-dien-9-amine, (18Z)-N,N-dimetylheptacos-18-en-10-amine, (17Z)-N,N-dimethylhexacos-17-en-9-amine, (19Z,22Z)-N,N-dimethyloctacosa-19,22-dien-7-amine, N,N-dimethylheptacos-an-10-amine, (20Z,23Z)-N-ethyl-N-methylnona-cosa-20,23-dien-10-amine, 1-[(11Z,14Z)-1-nonylicosa-11,14-dien-1-yl]pyrrolidine, (20Z)-N,N-dimethyl-heptacos-20-en-10-amine, (15Z)-N,N-dimethyl eptacos-15-en-10-amine, (14Z)-N,N-dimethylnonacos-14-en-10-amine, (17Z)-N,N-dimethylnonacos-17-en-10-amine, (24Z)-N,N-dimethyltritriacont-24-en-10-amine, (20Z)-N,N-di-methylnona-cos-20-en-10-amine, (22Z)-N,N-dimethyl-hen-triacont-22-en-10-amine, (16Z)-N,N-dimethylpenta-cos-16-en-8-amine, (12Z,15Z)-N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine, (13Z,16Z)-N,N-dimethyl-3-nonyldocos-a-13,16-dien-1-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclo-propyl]eptadec-an-8-amine, 1-[(1S,2R)-2-hexylcyclo-propyl]-N,N-dimethylnonadecan-10-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclo-propyl]nonadecan-10-amine, N,N-dimethyl-21-[R1S, 2R)-2-octylcyclopropyl]henicosan-10-amine,N,N-dimethyl-1-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]-methyl}cyclo-propyl]nonadecan-10-amine,N,N-dimethyl-1-[(1S,2R)-2-octylcyclo-propyl]hexadecan-8-amine, N,N-dimethyl-[(1R,2S)-2-undecyl-cyclopropyl]tetradecan-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-dimethyl-penta-decan-6-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclo-propyl]pentadecan-8-amine, R—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-(octyloxy)propan-2-amine, S—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-(octyloxy)propan-2-amine, 1-{2-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-1-[(octyloxy)methyl]ethyl}pyrrolidine, (2S)—N,N-dimethyl-1-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-3-[(5Z-)-oct-5-en-1-yloxy]propan-2-amine, 1-{2-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-1-[(octyloxy)methyl]ethyl}azetidine, (2S)-1-(hexyloxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine, (2S)-1-(heptyloxy)-N,N-di-methyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine, N,N-dimethyl-1-(non-yloxy)-3-[(9Z,12Z)-octadeca-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)-octa-deca-6,9,12-trien-1-yloxy]-3-(octyloxy)propan-2-amine, (2S)-1-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethyl-3-(pentyloxy)propan-2-amine, (2S)-1-(hexyloxy)-3-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethylpropan-2-amine, 1-[(11Z,14Z)-icosa-11,14-dien-1-yloxy]-N,N-dimethyl 1-3-(octyloxy)propan-2-amine, 1-[(13Z,16Z)-docosa-13,16-dien-1-yloxy]-N,N-dimethyl-3-(octyloxy)propan-2-amine, (2S)-1-[(13Z,16Z)-docosa-13,16-dien-1-yloxy]-3-(hexyloxy)-N,N-dimethyl-propan-2-amine, (2S)-1-[(13Z)-docos-13-en-1-yloxy]-3-(hexyloxy)-N,N-dimethyl propan-2-amine, 1-[(13Z)-docos-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-di-methyl-H(1-metoyloctyl)oxy]-3-[(9Z,12Z)-octa-deca-9,12-dien-1-yloxy]propan-2-amine, (2R)-1-[(3,7-dimethyloctyl)oxy]-N, N-dimethyl-3-R9Z,12Z)-octadeca-9,12-die-n-1-yloxyl-propan-2-amine, N,N-di-methyl-1-(octyloxy)-3-({8-[(1S,2S)-2-{[(1R,2R)-2-pentyl-cyclopropyl]-methyl}cyclopropyl]octyl}oxy)propan-2-amine, N,N-dimethyl-1-{[-(2-oclylcyclo-propyl)octyl]oxy}-3-(octyloxy)propan-2-amine and (11E,20Z,23Z)-N,N-dimethylnonacosa-11,20,2-trien-10-amine or a pharmaceutically acceptable salt or acid or stereoisomer thereof.
In one embodiment, the lipid may be a cleavable lipid such as those described in in Intl. Publ. No. WO2012/170889, which is herein incorporated by reference in its entirety
The cationic lipid can routinely be synthesized using methods known in the art (see, e.g., Intl. Publ. Nos. WO2012/040184, WO2011/153120, WO2011/149733, WO2011/090965, WO201/1043913, WO2011/022460, WO2012/061259, WO20121054365, WO2012/044638, WO2010/080724 and WO2010/21865; each of which is herein incorporated by reference in its entirety.
Lipid derivatives can include, for example, at least, the bonding (preferably covalent bonding) of one or more steric stabilizers and/or functional groups to the liposomal component after which the steric stabilizers and/or functional groups should be considered part of the liposomal components. Functional groups comprise groups that can be used to attach a liposomal component to another moiety such as a protein. Such functional groups include, at least, maleimide. These steric stabilizers include at least one from the group consisting of polyethylene glycol (PEG); poly-L-lysine (PLL); monosialo-ganglioside (GM1); poly(vinyl pyrrolidone) (PVP); poly(acrylamide) (PAA); poly(2-methyl-2-oxazoline); poly(2-ethyl-2-oxazoline); phosphatidylpoly-glycerol; poly[N-(2-hydroxy-propyl) methacrylamide]: amphiphilic poly-N-vinylpyrrolidones; L-amino-acid-based polymer; and polyvinyl alcohol.
In some embodiments, the provided trans-crocetin compositions are formulated in a lipid-polycation complex. The formation of the lipid-polycation complex may be accomplished using methods known in the art and/or as described in U.S. Pub. No. 2012/0178702, herein incorporated by reference in its entirety. As a non-limiting example, the polycation may include a cationic peptide or a polypeptide such as, but not limited to, polylysine, polyornithine and/or polyarginine and the cationic peptides described in International Pub. No. WO2012/013326; herein incorporated by reference in its entirety. In another embodiment, the provided trans-crocetin composition is formulated in a lipid-polycation complex which further includes a neutral lipid such as, but not limited to, cholesterol or dioleoyl phosphatidylethanolamine (DOPE).
Since the components of a liposome can include any molecule(s) (i.e., chemical/reagent/protein) that is bound to it, in some embodiments, the components of the provided liposomes include, at least, a member selected from: 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 liposomal components that make up the liposome comprises DSPE; DSPE-FITC; DSPE-maleimide; cholesterol; and HSPC.
In additional embodiments, the liposomes of the liposome compositions provided herein comprise oxidized phospholipids. In some embodiments, the liposomes comprise an oxidize phospholipid of a member selected from phosphatidylserines, phosphatidylinositols, phosphatidylethanolamines, phosphatidylcholines and 1-palmytoyl-2-arachidonoyl-sn-glycero-2-phosphate. In some embodiments, the phospholipids have unsaturated bonds. In some embodiments, the phospholipids are arachidonic acid containing phospholipids. In additional embodiments, the phospholipids are sn-2-oxygenated. In additional embodiments, the phospholipids are not fragmented.
In some embodiments, the liposomes of the disclosed liposome compositions comprise oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine (OxPAPC). The term “oxPAPC”, as used herein, refers to lipids generated by the oxidation of 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphorylcholine (PAPC), which results in a mixture of oxidized phospholipids containing either fragmented or full length oxygenated sn-2 residues. Well-characterized oxidatively fragmented species contain a five-carbon sn-2 residue bearing omega-aldehyde or omega-carboxyl groups. Oxidation of arachidonic acid residue also produces phospholipids containing esterified isoprostanes. oxPAPC includes HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC species, among other oxidized products present in oxPAPC. In further embodiments, the oxPAPCs are epoxyisoprostane-containing phospholipids. In further embodiments, the oxPAPC is 1-palmitoyl-2-(5,6-epoxyisoprostane E2)-sn-glycero-3-phosphocholine (5,6-PEIPC), 1-palmitoyl-2-(epoxy-cyclopenten-one)-sn-glycero-3-phosphoryl-choline (PECPC) and/or 1-palmitoyl-2-(epoxy-isoprostane E2)-sn-glycero-4-phosphocholine (PEIPC). In some embodiments, the phospholipids have unsaturated bonds. In some embodiments, the phospholipids are arachidonic acid containing phospholipids. In additional embodiments, the phospholipids are sn-2-oxygenated. In additional embodiments, the phospholipids are not fragmented.
In some embodiments, the liposomes of the disclosed liposome compositions comprise a lipid selected from: 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC); 1-palmitoyl-2-(9′oxo-nonanoyl)-sn-glycero-3-phospho-choline; 1-palmitoyl-2-arachinodoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phos-phocholine; 1-palmitoyl-2-hexadec-yl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azel-aoyl-sn-glycero-3-phos-phocholine; and 1-palmitoyl-2-acetoyl-sn-glycero-3-phospho-choline. In further embodiments, the liposome comprises PGPC.
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 that comprises a reactive group that can be used to crosslink reagents and moieties to the lipid. If the lipid is functionalized, any liposome that it forms is also functionalized. In some embodiments, the reactive group is one that will react with a crosslinker (or other moiety) to form crosslinks. The reactive group in the liposome lipid bilayer is located anywhere on the lipid that allows it to contact a crosslinker and be crosslinked to another moiety (e.g., a steric stabilizer or targeting moiety). In some embodiments, the reactive group is in the head group of the 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 dithiol crosslinkers including but not limited to dithiothreitol (DTT).
It is to be understood that the use of other functionalized lipids, other reactive groups, and other crosslinkers beyond those described above is further contemplated. In addition to the maleimide groups, other examples of contemplated reactive groups include but are not limited to other thiol reactive groups, amino groups such as primary and secondary amines, carboxyl groups, hydroxyl groups, aldehyde groups, alkyne groups, azide groups, carbonyls, halo acetyl (e.g., iodoacetyl) groups, imidoester groups, N-hydroxysuccinimide esters, sulfhydryl groups, and pyridyl disulfide groups.
Functionalized and non-functionalized lipids are available from a number of commercial sources including Avanti Polar Lipids (Alabaster, AL) and Lipoid LLC (Newark, NJ).
In some embodiments, the liposomes include a steric stabilizer that increases their longevity in circulation. One or more steric stabilizers such as a hydrophilic polymer (polyethylene glycol (PEG)), a glycolipid (monosialo-ganglioside (GM1)) or others occupies the space immediately adjacent to the liposome surface and excludes other macromolecules from this space. Consequently, access and binding of blood plasma opsonins to the liposome surface are hindered, and thus interactions of macrophages with such liposomes, or any other clearing mechanism, are inhibited and longevity of the liposome in circulation is enhanced. In some embodiments, the steric stabilizer or the population of steric stabilizers is a PEG or a combination comprising PEG. In further embodiments, the steric stabilizer is a PEG or a combination comprising PEG with a number average molecular weight (Mn) of 200 to 5000 Daltons. These PEG(s) can be of any structure such as linear, branched, star or comb structure and are commercially available.
In some embodiments, liposomes of the provided liposomal compositions are pegylated (e.g., pegylated liposomal CTC and pegylated liposomal MTC). In some embodiments, the pegylated liposomes are water soluble. That is, the pegylated liposomes are in the form of an aqueous solution.
The diameter of the provided liposomes is not particularly limited. In some embodiments, the liposomes have a mean diameter of for example, 20 nm to 500 nm (nanometer), or 20 nm to 200 nm, or any range therein between. In some embodiments, the liposomes have a mean diameter of 80 nm to 120 nm, or any range therein between.
In some embodiments, the pH of solutions comprising the liposome composition is from pH 2 to 8, or any range therein between. In some embodiments, the pH of solutions comprising the liposome composition is from pH 5 to 8, or 6 to 7, or any range therein between. In some embodiments, the pH of solutions comprising the liposome composition is from pH 6 to 7, or any range therein between. In some embodiments, the pH of solutions comprising the liposome composition is from 6 to 7.5, from 6.5 to 7.5, from 6.7 to 7.5, or from 6.3 to 7.0, or any range therein between.
In additional embodiments, the provided liposomal composition comprises a buffer. In further embodiments, the buffer is selected from HEPES, citrate, or sodium phosphate (e.g., monobasic and/or dibasic sodium 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., monobasic and/or dibasic sodium phosphate). In some embodiments, the buffer is at a concentration of 15 to 200 mM, or any range therein between. In yet further embodiments, the buffer is at a concentration of 5 to 200 mM, 15 to 200, 5 to 100 mM, 15 to 100 mM, 5 to 50 mM, 15 to 50 mM, 5 to 25 mM, 5 to 20 mM, 5 to 15 mM, or any range therein between. In some embodiments, the buffer is HEPES at a concentration of 5 to 200 mM, or any range therein between. In some embodiments, the buffer is citrate at a concentration of 5 to 200 mM, or any range therein between. In some embodiments, the buffer is sodium phosphate at a concentration of 5 to 200 mM, or any range therein between.
In additional 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 at 1 to 20%, or 5 to 20% weight percent, or any range therein between.
In additional embodiments, the provided liposomal composition 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 therein between. In some embodiments, the liposome composition includes a 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 therein between.
In some embodiments, the provided liposomal composition comprises trehalose. In further embodiments, the concentration weight percent of trehalose is 0.1-20%, 1-20%, 0.5-15%, 1%-15%, 5-20%, or 1-50%, or any range therein between. In yet further embodiments, the concentration (weight percent) of trehalose is 1-15%, or any range therein between. In an additional embodiment, the trehalose is present at about 5% to 20% weight percent of trehalose or any combination of one or more lyoprotectants or cryoprotectants at a total concentration of 5% to 20%. In some embodiments, the pH of the liposome composition is from 6 to 7.5, from 6.5 to 7.5, from 6.7 to 7.5, or from 6.3 to 7.0, or any range therein between.
In some embodiments, the liposome composition comprises dextrose. In some embodiments, the concentration weight percent of dextrose is 0.1-20%, 1-20%, 0.5-15%, 1-15%, 5-20%, or 1-50%, or any range therein between. In particular embodiments, the concentration (weight percent) of dextrose is 1-20%, or any range therein between. In an additional embodiment, the dextrose is present at 1 to 20% weight percent of dextrose or any combination of one or more lyoprotectants or cryoprotectants at a total concentration of 1% to 20%, or 5% to 20%, or any range therein between.
In some embodiments, the disclosure provides a liposome composition that comprises a liposome encapsulating a trans-crocetin salt. 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 nontargeted. 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, molecules of trans-crocetin. In some embodiments, the liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000 molecules, or any range therein between. In some embodiments, the liposome encapsulates trans-crocetin and one or more of different carotenoids. In some embodiments, the unpegylated and targeted liposome contains 10-50, 10 to 100, 25 to 75, or 30 to 200 targeting moieties, or any range therein between.
In further embodiments, the provided liposomal compositions comprise a liposome encapsulating a trans-crocetin 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% weight of trehalose. In some embodiments, the liposome solution contains HBS at a concentration of 1 to 200 mM and a pH of 2-8, or any range therein between. In some embodiments, liposome solution has a pH 5-8, or any range therein between. In some embodiments, liposome solution has a pH 6-7, or any range therein between. In some embodiments, the provided trans-crocetin salt is a multivalent salt (e.g., divalent, trivalent, or tetravalent). In some embodiments, the trans-crocetin salt is CTC. In some embodiments, the trans-crocetin salt is MTC.
The provided liposomes comprise an aqueous compartment enclosed by at least one lipid bilayer. When lipids that include a hydrophilic headgroup are dispersed in water they can spontaneously form bilayer membranes referred to as lamellae. The lamellae are composed of two monolayer sheets of lipid molecules with their non-polar (hydrophobic) surfaces facing each other and their polar (hydrophilic) surfaces facing the aqueous medium. The term liposome includes unilamellar vesicles which are comprised of a single lipid bilayer and generally have a diameter of about 20 to about 500 nm, about 50 to about 300 nm, about 50 to about 150 nm, about 30 to about 1000 nm, about 30 to about 175 nm, about 80 to about 400 nm, or about 80 to about 120 nm. Liposomes can also be multilamellar, which generally have a diameter 0.5 to 10 μm with anywhere from two to hundreds of concentric lipid bilayers alternating with layers of an aqueous phase. In some embodiments, liposomes can include multilamellar vesicles (MLV), large unilamellar vesicles (LUV), and small unilamellar vesicles (SUV). The lipids of the liposome can be cationic, zwitterionic, neutral or anionic, or any mixture thereof.
The size of the liposomes in the provided liposomal compositions may vary from for example, 0.5 nm to 10 μm, or 20 nm to 5 um, depending on the phospholipid composition, the method used for their preparation, and the intended therapeutic use of the liposomes. In some embodiments, the median diameter of the liposomes in the provided liposomal composition is 20 nm to 500 nm, 50 nm to 200 nm, or 20 nm to 200 nm, or any range therein between. In some embodiments, the liposome median diameter is 80 nm to 120 nm, or any range therein between (e.g., 85-115 nm, 90-110 nm, 95-110 nm, or 95-105 nm). In some embodiments, the median diameter of the liposomes in the provided liposomal composition is 10-250 nm, or any range therein between (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 mu, 10-125 nm, 10-100 nm, 10-75 nm, 10-50 nm, 50-100 nm, 50-90 nm, 50-80 nm, 50-70 nm, 50-60 nm, 60-100 nm, 60-90 nm, 60-80 nm, 60-70 nm, 70-100 nm, 70-90 nm, 70-80 nm, 80-100 nm, 80-90 nm, or 90-100 nm). In some embodiments, the median diameter of the liposomes in the provided liposomal composition is 100-250 nm, or any range therein between (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 provided liposomal composition is 10-100 nm, or any range therein between (e.g., from 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 provided liposomal composition 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 light scattering is a method used to measure the diameter of liposomes that is well known to those skilled in the art. The diameter of the liposomes (DLP) can routinely be determined using any techniques and equipment known in the art including for example, dynamic laser light scattering (Coulter N4 particle size analyzer), the Zetasizer Nano ZSP (Malvern, UK), and an ELS-8000 (Otsuka Electronics Co., Ltd.)).
In some embodiments, the provided liposomal compositions have a monodisperse size (diameter) distribution. “Monodisperse” and “homogeneous size distribution,” are used interchangeably herein and describe a plurality of liposomal nanoparticles or microparticles where the particles have the same or nearly the same diameter. As used herein, a monodisperse distribution refers to particle distributions in which 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 86%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% or greater of the particle distribution lies within 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, or 10% of the mass median diameter.
In some embodiments, the liposome population in the provided liposomal composition is relatively homogenous. In some embodiments, the liposome population in the provided liposomal composition is heterogeneous. A polydispersity index may be used to indicate the homogeneity of a nanoparticle composition, e.g., the particle size (diameter) distribution of the nanoparticle compositions. A small (e.g., less than 0.3) polydispersity index generally indicates a narrow particle size distribution. In some embodiments, the liposome population in the provided liposomal composition has a polydispersity index from 0 to 0.25, or 0.01 to 0.1, or any range therein between (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 therein between.
In some embodiments, liposomes in the liposome population in the provided liposomal composition differ in their lipid composition, molar ratio of lipid components, size, charge (zeta potential), targeting ligands and/or combinations thereof.
The zeta potential of a nanoparticle composition may be used to indicate the electrokinetic potential of the composition. For example, the zeta potential may describe the surface charge of a nanoparticle composition. Nanoparticle compositions with relatively low charges, positive or negative, are generally desirable, as more highly charged species may interact undesirably with cells, tissues, and other elements in the body. In some embodiments, the zeta potential of a nanoparticle composition can be from about −10 mV to about +20 mV, from about −10 mV to about +15 mV, from about −10 mV to about +10 mV, from about −10 mV to about +5 mV, from about −10 mV to about 0 mV, from about −10 mV to about −5 mV, from about −5 mV to about +20 mV, from about −5 mV to about +15 mV, from about −5 mV to about +10 mV, from about −5 mV to about +5 mV, from about −5 mV to about 0 mV, from about 0 mV to about +20 mV, from about 0 mV to about +15 mV, from about 0 mV to about +10 mV, from about 0 mV to about +5 mV, from about +5 mV to about +20 mV, from about +5 mV to about +15 mV, or from about +5 mV to about +10 mV. Liposome zeta potential can routinely be determined using techniques and equipment known in the art including for example, dynamic light scattering (Zetasizer Nano ZSP, Malvern, UK) and laser Doppler electrophoresis.
The encapsulation efficiency of a therapeutic and/or prophylactic such as trans-crocetin, describes the amount of therapeutic and/or prophylactic that is encapsulated or otherwise associated with a nanoparticle composition after preparation, relative to the initial amount provided. The encapsulation efficiency is desirably high (e.g., close to 100%). The encapsulation efficiency may be measured, for example, by comparing the amount of therapeutic and/or prophylactic in a solution containing the nanoparticle composition before and after removing the unencapsulated therapeutic and/or prophylactic drug. For the liposome compositions described herein, the encapsulation efficiency of trans-crocetin can be at least 50%, for example 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%.
In additional embodiments, the provided liposomal compositions contain liposomes encapsulating a trans-crocetin salt. In some embodiments, the trans-crocetin/lipid ratio of the provided liposomal composition is 1 to 1000 g/mol, or any range therein between. In some embodiments, the trans-crocetin/lipid ratio of the liposome composition is 10 to 200 g/mM, 10 to 150 g/mM, 10 to 100 g/mM, 20 to 200 g/mM, 20 to 150 g/mM, 20 to 100 g/mM, 30 to 200 g/mM, 30 to 150 g/mM, 30 to 100 g/mM, 40 to 200 g/mM, 40 to 150 g/mM, 40 to 100 g/mM, 50 to 200 g/mM, 50 to 150 g/mM, or 50 to 100 g/mM, or any range therein between. In some embodiments, trans-crocetin/lipid ratio is 30 to 90 g/mM, or any range therein between. In some embodiments, trans-crocetin/lipid ratio is 30 to 50 g/mM, 40 to 60 g/mM, 50 to 70 g/mM, 60 to 80 g/mM, or 70 to 90 g/mM, or any range therein between. In additional embodiments, the trans-crocetin/lipid ratio of the liposome composition is 20 to 120 g/mM (e.g., about 25 to 100 g/mM), or any range therein between.
In some embodiments, the liposome composition is buffered using a zwitterionic buffer. Suitably, the zwitterionic buffer is an aminoalkanesulfonic acid or suitable salt. Examples of aminoalkanesulfonic 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 in for use in a commercial injection product. Most preferably the buffer is HEPES. The liposome composition may suitable include an AGP.
In some embodiments, the liposome composition is buffered using HEPES. In some embodiments, the liposome composition is buffered using HEPES having a pH of about 7.
In some embodiments, the pharmaceutical composition is a liposome composition comprising a cationic liposome. In some embodiments, the liposome composition comprises a liposome that has a zeta potential that is more 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 therein between. In some embodiments, the liposome has a diameter of 20 nm to 500 nm, 20 nm to 200 nm, 30 nm to 175 nm, 50 nm to 200 nm, or 50 nm to 150 nm, or any range therein between. In some embodiments, the cationic liposome has a diameter of 80 nm to 120 nm, or any range therein between. 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 trans-crocetin. In some embodiments, during the process of preparing 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 liposomal trans-crocetin starting material is encapsulated (entrapped) in the liposomes of the liposome composition. In additional embodiments, trans-crocetin encapsulated by the liposome is in a HEPES buffered solution within the liposome. In further embodiments, the liposome comprises at least one OxPAPC.
In some embodiments, the provided pharmaceutical composition is a liposome composition comprising an anionic or neutral liposome. In some embodiments, the liposome composition comprises a liposome that has a zeta potential that is less than or equal to zero. In some embodiments, the liposome has a zeta potential that 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 therein between. In some embodiments, the anionic or neutral liposome has a diameter of 20 nm to 500 nm, 20 nm to 200 nm, 30 nm to 175 nm, or 50 nm to 150 nm, or any range therein between. In other embodiments, the anionic or neutral liposome has a diameter of 80 nm to 120 nm, or any range therein between. In some embodiments, the anionic liposome has a diameter of 20 nm to 500 nm, 20 nm to 200 nm, 30 nm to 175 nm, or 50 nm to 150 nm, or any range therein between. In further embodiments, the anionic liposome has a diameter of 80 nm to 120 nm, or any range therein between. In some embodiments, the neutral liposome has a diameter of 20 nm to 500 nm, 20 nm to 200 nm, 30 nm to 175 nm, or 50 nm to 150 nm, or any range therein between. In some embodiments, the neutral liposome has a diameter of 80 nm to 120 nm, or any range therein between. 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 more than 75%, w/w trans-crocetin. In some embodiments, during the process of preparing the liposome composition, at least 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, or more than 75%, of the starting material of trans-crocetin is encapsulated (entrapped) in the liposomes. 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 the 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 the trans-crocetin. In some embodiments, 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 the trans-crocetin. In additional embodiments, the trans-crocetin is encapsulated by the anionic or neutral liposome is in a HEPES buffered solution within the liposome. In further embodiments, the liposome comprises at least one OxPAPC.
In some embodiments, the provided pharmaceutical composition is a liposome composition comprising a liposome that comprises at least one OxPAPC. In some embodiments, the OxPAPC is an oxidized and/or phospholipid containing fragmented oxygenated sn-2 residues. In some embodiments, the OxPAPC is an oxidized phospholipid containing a five-carbon sn-2 residue bearing an omega-aldehyde or omega-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 epoxyisoprostane-containing phospholipid. In some embodiments, the OxPAPC is PGPC. In some embodiments, the liposome comprises at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30%, OxPAPC. In some embodiments, the liposome composition has a cationic liposome that comprises 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, OxPAPC, or any range therein between. In some embodiments, the liposome composition comprises a cationic liposome. In some embodiments, the liposome composition comprises a neutral liposome. In some embodiments, the liposome composition comprises an anionic liposome. In additional embodiments, the liposome composition comprises at least one liposome containing an OxPAPC that has a diameter of 20 nm to 500 nm, 20 nm to 200 nm, 30 nm to 175 nm, or 50 nm to 150 nm, or any range therein between. In further embodiments, the liposome composition comprises a at least one liposome containing an OxPAPC that has a diameter of 80 nm to 120 nm, or any range therein between.
In some embodiments, the provided pharmaceutical composition is a liposome composition comprising a cationic liposome that comprises at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%,25%, or at least 30%, OxPAPC. In some embodiments, the liposome composition has a cationic liposome that comprises 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, OxPAPC, or any range therein between. In some embodiments, the liposome comprises at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30%, OxPAPC. In some embodiments, the liposome composition has a cationic liposome that contains about 10% OxPAPC. In some embodiments, the liposome composition has a cationic liposome that comprises at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30%, PGPC. In some embodiments, the liposome comprises 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, PGPC, or any range therein between. In some embodiments, the liposome composition has a cationic liposome that comprises about 10% PGPC.
In some embodiments, the pharmaceutical composition is a liposome composition comprising an anionic or neutral liposome that comprises at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30%, OxPAPC. In some embodiments, the liposomal composition has a anionic or neutral liposome that comprises 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, OxPAPC, or any range therein between. In some embodiments, the liposome comprises at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30%, OxPAPC. In some embodiments, the liposomal composition has a anionic or neutral liposome that contains about 10% OxPAPC. In some embodiments, the liposomal composition comprises has a anionic or neutral liposome that comprises at least 0.01%, 0.1%, 1%, 5%, 10%, 15%,20%, 25%, or at least 30%, PGPC. In some embodiments, the liposome comprises 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, PGPC, or any range therein between. In some embodiments, the liposomal composition has a anionic or neutral liposome that contains about 10% PGPC.
In some embodiments, the pharmaceutical composition is a liposomal composition comprising a neutral liposome that comprises at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30%, OxPAPC. In some embodiments, the neutral OxPAPC containing liposomal composition comprises 0.01%-35%, 0.1%-30%, 1%-25%, 3-20%, or 5-15%, OxPAPC, or any range therein between. In some embodiments, the neutral OxPAPC containing liposomal composition comprises about 10% OxPAPC. In some embodiments, the neutral OxPAPC containing liposomal composition comprises at least 0.01%, 0.1%, 1%, 5%, 10%, 15%, 20%, 25%, or at least 30%, PGPC. In some embodiments, the neutral PGPC containing liposomal composition comprises 0.01%-35%, 0.1%-30%, 1%-25%, 3%-20%, or 5-15%, PGPC, or any range therein between. In some embodiments, the neutral OxPAPC containing liposomal composition comprises about 10% PGPC.
In some embodiments, the pharmaceutical composition is a liposomal composition comprising a surface active copolymer. Surface active copolymers, also termed block polymer nonionic surfactants, am surface active agents synthesized by the sequential addition of two or more alkylene oxides to a low molecular weight water soluble organic compound containing one or more active hydrogen atoms. In some embodiments, the liposomal composition comprises a surface active copolymer selected from a poloxamer, meroxapol, poloxamine, and PLURADOT™. The surface active copolymers in the liposomal composition can be encapsulated by, or integrated into or otherwise attached with the liposomes by covalent, ionic, or other binding interaction and/or the surface active copolymers may not be encapsulated by, integrated into or otherwise attached with liposomes in the liposomal composition (e.g., the surface active copolymers may be free in the liposomal composition).
In some embodiments, the liposomal composition comprises a poloxamer such as P188, and P124, P182, P188, and P234, have been reported to bind to cell membranes and markedly reduce cell permeability that has been induced by ischemic injury. The embodiments described herein also deliver increased oxygen to the organs and cells more effectively, and in a way that reduces reperfusion injury. Without wishing to be limited to any particular theory or mechanism, it is believed that this oxygen delivery reduces the intracellular injury that is attributable to mitochondrial dysfunction and/or metabolic and enzymatic abnormalities associated with poor perfusion and/or reperfusion injury. In some embodiments, the liposomal composition comprises a poloxamer with a molecular weight of between 2,000 and 20,000 Daltons. Poloxamers within this molecular weight range remain soluble in water while minimizing potential toxicity. In some embodiments, the poloxamer's hydrophobic group has a molecular weight range from approximately 950-4,000 Daltons. In such embodiments, the hydrophilic groups may constitute approximately 45-95% by weight of the poloxamer. In an exemplary embodiment, the hydrophobic group has a molecular weight of 1,750-3,500 Daltons and the hydrophilic groups constitute between 50-90% by weight of the molecule.
In some embodiments, the liposomal composition comprises at least one poloxamer selected from P108, P124, P138, P171, P181, P182, P185, P188, P234, P237, P288, and P407. In some embodiments, the liposomal composition comprises at least one poloxamer selected from P124, P182, P188, and P234.
In particular embodiments, the liposomal composition comprises poloxamer 188 (P188) (Pluronic F68).
In additional embodiments, a liposome in the liposomal composition is pegylated.
In some embodiments, the provided pharmaceutical composition is a non-targeted liposomal composition. That is, the liposomes in the liposomal composition do not have specific affinity towards an epitope (e.g., an epitope on a surface antigen) expressed on the surface of a target cell of interest. In further embodiments, the non-targeted liposomal composition is pegylated.
In some cases, liposome accumulation at a target site may be due to the enhanced permeability and retention characteristics of certain tissues such as cancer tissues. Accumulation in such a manner often results in part because of liposome size and may not require special targeting functionality. In other embodiments, the provided liposomes include a targeting agent. Generally, the targeting agents can associate with any target of interest, such as a target associated with an organ, tissues, cell, extracellular matrix, or intracellular region. In certain embodiments, a target can be associated with a particular disease state, such as a cancerous condition. In some embodiments, the targeting component can be specific to only one target, such as a receptor. Suitable targets can include but are not limited to a nucleic acid, such as a DNA, RNA, or modified derivatives thereof. Suitable targets can also include but are not limited to a protein, such as an extracellular protein, a receptor, a cell surface receptor, a tumor-marker, a transmembrane protein, an enzyme, or an antibody. Suitable targets can include a carbohydrate, such as a monosaccharide, disaccharide, or polysaccharide that can be, for example, present on the surface of a cell.
In certain embodiments, a targeting agent can include a target ligand (e.g., an RGD-containing peptide), a small molecule mimic of a target ligand (e.g., a peptide mimetic ligand), or an antibody or antibody fragment specific for a particular target. In some embodiments, a targeting agent can further include folic acid derivatives, B-12 derivatives, integrin RGD peptides, NGR derivatives, somatostatin derivatives or peptides that bind to the somatostatin receptor, e.g., octreotide and octreotate, and the like. In some embodiments, the targeting agents include an aptamer. Aptamers can be designed to associate with or bind to a target of interest. Aptamers can be comprised of, for example, DNA, RNA, and/or peptides, and certain aspects of aptamers are known in the art. (See, e.g., Klussman, Ed., The Aptamer Handbook, Wiley-VCH (2006); Nissenbaum, Trends in Biotech. 26(8): 442-449 (2008)).
In other embodiments, the liposomal composition comprises a targeted liposome. That is, the liposome contains a targeting moiety that has 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 through a covalent bond. In other embodiments, the targeting moiety of the liposome is attached to one or both of a PEG and the exterior of the liposome. In further embodiments, the targeted liposome is pegylated. The functions of the targeting moiety of the targeted liposome may include but is not limited to, targeting the liposome to the target cell of interest in vivo or in vitro; interacting with the surface antigen for which the targeting moiety has specific affinity, and delivering the liposome payload (e.g., trans-crocetin) to the location of or into the cell. In some embodiments, the target cell of interest is a cell/tissue of the blood brain barrier. In some embodiments, the target cell of interest expresses transferring receptor In some embodiments, the target cell of interest expresses transferrin receptor and the targeting moiety exhibits rapid dissociation at endosomal pH, while maintaining a high affinity for TfR at neutral pH. In some embodiments, the targeting moiety has affinity for a cell surface antigen that is abundantly expressed. In some embodiments, the targeting moiety has affinity for a cell surface antigen that is abundantly expressed in the microvasculature. In some embodiments, the targeting moiety has affinity for the cell surface antigen EphA2.
In some embodiments, the administered liposomes contains a targeting moiety selected from: an antibody, a small molecule, a polypeptide, an aptamer, a folate, a transferrin (e.g., lactoferrin), a glycoprotein, an integrin, glutathione, and a carbohydrate. In some embodiments, administered liposomes contains 10-50, 10 to 100, 25 to 75, or 30 to 200 targeting moieties.
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 that comprises 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, and an antigen binding fragment of an antibody, a single chain antibody, a single-domain antibody, a bi-specific antibody, a synthetic antibody, a pegylated antibody, and a multimeric antibody. In some embodiments, the targeting moiety has specific affinity for an epitope that is preferentially expressed on a target cell such as a tumor cell, compared to normal or non-tumor cells. In some embodiments, the targeting moiety has specific affinity for an epitope on a tumor cell surface antigen that is present on a tumor cell but absent or inaccessible on a non-tumor cell. In some embodiments, the targeting moiety binds an epitope of interest with an equilibrium dissociation constant (Kd) in a range of 0.5×10−6 to 10×10−4, 0.5×10−10 to 10×10−6, or 50×10−12 to 10×10−6 as determined using BIACORE® analysis. In further embodiments, the Kd is determined using a surface plasmon resonance technique in which an antigen containing the epitope is immobilized, the targeting moiety serves as analyte, and the following conditions are used: 10 mM MES buffer, 0.05% polyoxyethylene sorbitan monolaurate, and 150 mM NaCl at 37° C.
In additional embodiments, the liposome composition comprises one or more of an immunostimulatory agent, a detectable marker, and a maleimide, disposed on at least one of the PEG and the exterior of the liposome. In some embodiments, a liposome of the liposome composition is cationic. In other embodiments, a liposome of the liposome composition is anionic or neutral. In additional embodiments, a liposome of the liposomal composition has a diameter of 20 nm to 500 nm, or any range therein between. In further embodiments, a liposome of the liposomal composition has a diameter of 80 nm to 120 nm, or any range therein between. In some embodiments, a liposome of the liposomal composition is pegylated. In some embodiments, a liposome of the liposomal composition is targeted. In further embodiments, a liposome of the liposomal composition is pegylated and targeted.
In some embodiments, the pharmaceutical composition comprises a trans-crocetin salt having the formula: Q-trans-crocetin-Q
In some embodiments, the pharmaceutical composition administered according to the provided methods is a fixed dose of liposomal trans-crocetin. In some embodiments, the administered liposomal trans-crocetin composition does not contain a fixed dose of liposomal trans-crocetin.
In some embodiments, Q is a multivalent cation 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 cation counterion. In further embodiments, Q is a divalent metal cation. In some embodiments, Q is at least one member selected from Ca2+, Mg2+, Zn2+, Cu2+, Co2+, and Fe2+. In further embodiments, Q is Ca2+ or Mg2+. In some embodiments, Q is Ca2+. In some embodiments, Q is Mg2+. In some embodiments, Q is a divalent organic counterion. In other embodiments, Q is a trivalent cation counterion such as Fe3+. In other embodiments, Q is a multivalent organic counterion. In some embodiments, Q is a divalent organic cation. In some embodiments, Q is a bivalent organic cation such as protonated diamine.
In further embodiments, Q is a monovalent cation counterion. In some embodiments, Q is a monovalent metal cation. In some embodiments, Q is at least one member 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+, a protonated diamine or a protonated polyamine.
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, molecules of trans-crocetin. In some embodiments, the liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, molecules of trans-crocetin, or any range therein between. In some embodiments, the trans-crocetin/lipid ratio of the liposomal composition is 1 g/mol and about 1000 g/mol, or any range therein between. 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 therein between. In some embodiments, the liposome comprises at least 0.1% to 97% trans-crocetin. In some embodiments, the liposome has a diameter of 20 nm to 500 nm, or 20 nm to 200 nm, or any range therein between. In some embodiments, the liposome has a diameter of 80 nm to 120 nm, or any range therein between. In some embodiments, the liposome is formed from liposomal components. In further embodiments, the liposomal components comprise at least one of an anionic lipid and a neutral lipid. In further embodiments, the liposomal components comprise at least one selected from: DSPE; DSPE-PEG; DSPE-PEG-maleimide; HSPC; HSPC-PEG; cholesterol; cholesterol-PEG; and cholesterol-maleimide. In further embodiments, the liposomal components comprise at least one selected from: DSPE; DSPE-PEG; DSPE-PEG-FITC; DSPE-PEG-maleimide; cholesterol; and HSPC. In additional embodiments, the liposome further comprises an oxidized phospholipid such as an OxPAPC. In some embodiments, the liposome comprises an OxPAPC that 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 a five-carbon sn-2 residue bearing omega-aldehyde or omega-carboxyl groups. In some embodiments, the liposome comprises an OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC, or the OxPAPC is an epoxyisoprostane-containing phospholipid. In some embodiments, the liposome comprises an OxPAPC selected from 1-palmitoyl-2-(5,6-epoxyisoprostane E2)-sn-glycero-3-phosphocholine (5,6 PEIPC), 1-palmitoyl-2-(epoxy-cyclopenten-one)-sn-glycero-3-phosphorylcholine (PECPC), 1-palmit-oyl-2-(epoxyisoprostane E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC); 1-palmitoyl-2-(9′oxo-nonan-oyl)-sn-glycer-o-3-phosphocholine; 1-palmitoyl-2-ar-achinodoyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine; 1-palmit-oyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetoyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, the OxPAPC within the liposome lipid bilayer is 0%-100% of total lipids, or any range therein between. In some embodiments, the liposome comprises a targeting moiety having a specific affinity for a surface antigen on a target cell of interest (e.g., an ehprin receptor such as EphA2 or transferrin receptor). In some embodiments, the targeting moiety is attached to one or both of a PEG and the exterior of the liposome, optionally wherein the targeting moiety is attached to one or both of the PEG and the exterior of the liposome by a covalent bond. 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 liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200 targeting moieties, or any range therein between. In some embodiments, the liposome further comprises an immunostimulating agent (such as 1,6-beta 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 additional embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential that is less than or equal to zero. In some embodiments, the liposome has a zeta potential that 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 therein between. In other embodiments, the liposome is cationic. In some embodiments, the liposomal composition comprises a liposome that has a zeta potential that is more than zero. In some embodiments, the liposome has a zeta potential that is 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 therein between.
In some embodiments, the disclosure provides a pharmaceutical composition comprising calcium trans-crocetinate (CTC) encapsulated by a liposome. The CTC can exist in linear and/or cyclic form (shown below).
In some embodiments, the pharmaceutical composition administered according to the provided methods is a fixed dose of liposomal CTC. In some embodiments, the administered liposomal trans-crocetin composition does not contain a fixed dose of liposomal CTC.
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, molecules of trans-crocetin. In some embodiments, the liposome contains 10 to 100,000, 100 to 10,000, or 1,000 to 5,000, molecules, or any range therein between. In some embodiments, the trans-crocetin/lipid ratio of the liposomal composition is 1 g/mol and about 1000 g/mol, or any range therein between. 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 therein between. In some embodiments, the liposome comprises at least 0.1% to 97% trans-crocetin. In some embodiments, the liposome has a diameter of 20 nm to 500 nm, or 20 nm to 200 nm, or any range therein between. In some embodiments, the liposome has a diameter of 80 nm to 120 nm, or any range therein between. In some embodiments, the liposome is formed from liposomal components. In further embodiments, the liposomal components comprise at least one of an anionic lipid and a neutral lipid. In further embodiments, the liposomal components comprise at least one selected from: DSPE; DSPE-PEG; DSPE-PEG-maleimide; HSPC; HSPC-PEG; cholesterol; cholesterol-PEG; and cholesterol-maleimide. In further embodiments, the liposomal components comprise at least one selected from: DSPE; DSPE-PEG; DSPE-PEG-FITC; DSPE-PEG-maleimide; cholesterol; and HSPC. In additional embodiments, the liposome further comprises an oxidized phospholipid such as an OxPAPC. In some embodiments, the liposome comprises an OxPAPC that 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 a five-carbon sn-2 residue bearing omega-aldehyde or omega-carboxyl groups. In some embodiments, the liposome comprises an OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC, or the OxPAPC is an epoxyisoprostane-containing phospholipid. In some embodiments, the liposome comprises an OxPAPC selected from 1-palmitoyl-2-(5,6-epoxyisoprostane E2)-sn-glycero-3-phosphocholine (5,6 PEIPC), 1-palmitoyl-2-(epoxy-cyclopenten-one)-sn-glycero-3-phosphorylcholine (PECPC), 1-palmit-oyl-2-(epoxy-isoprostane E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC); 1-palmitoyl-2-(9′oxononanoyl)-sn-glycero-3-phosphocholine; 1-palmitoyl-2-arachinodoyl-sn-glycero-3-phospho-choline; 1-palmitoyl-2-myristoyl-sn-glycer-o-3-phosphocholine; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetoyl-sn-glycero-3-phospho-choline. In some embodiments, the liposome comprises PGPC. In some embodiments, the OxPAPC within the liposome lipid bilayer is 0%-100% of total lipids, or any range therein between. 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 a PEG and the exterior of the liposome, optionally wherein the targeting moiety is attached to one or both of the PEG and the exterior of the liposome by a covalent bond. 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 liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200 targeting moieties, or any range therein between. In some embodiments, the liposome contains less than 500,000 or less than 200,000 molecules of trans-crocetin. In some embodiments, the liposome contains between 10 to 100,000 molecules, or any range therein between. In some embodiments, the liposome further comprises an immunostimulating agent (such as 1,6-beta 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 additional embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential that is less than or equal to zero. In some embodiments, the liposome has a zeta potential that 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 therein between. In other embodiments, the liposome is cationic. In some embodiments, the liposomal composition comprises a liposome that has a zeta potential that is more than zero. In some embodiments, the liposome has a zeta potential that is 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 therein between.
In some embodiments, the disclosure provides a pharmaceutical composition comprising magnesium trans-crocetinate (MTC) encapsulated by a liposome. The MTC can exist in linear and/or cyclic form (shown below).
In some embodiments, the pharmaceutical composition administered according to the provided methods is a fixed dose of liposomal CTC. In some embodiments, the administered liposomal trans-crocetin composition does not contain a fixed dose of liposomal CTC.
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, molecules of trans-crocetin. In some embodiments, the liposome contains 10 to 100,000, 100 to 10,000, or 500 to 5,000, molecules, or any range therein between. 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 therein between. In some embodiments, the liposome comprises at least 0.1% to 97% trans-crocetin. In some embodiments, the liposome has a diameter of 20 nm to 500 nm, or 20 nm to 200 nm, or any range therein between. In some embodiments, the liposome has a diameter of 80 nm to 120 nm, or any range therein between. In some embodiments, the liposome is formed from liposomal components. In further embodiments, the liposomal components comprise at least one of an anionic lipid and a neutral lipid. In further embodiments, the liposomal components comprise at least one selected from: DSPE; DSPE-PEG; DSPE-PEG-maleimide; HSPC; HSPC-PEG; cholesterol; cholesterol-PEG; and cholesterol-maleimide. In further embodiments, the liposomal components comprise at least one selected from: DSPE; DSPE-PEG; DSPE-PEG-F1TC; DSPE-PEG-maleimide; cholesterol; and HSPC. In additional embodiments, the liposome further comprises an oxidized phospholipid such as an OxPAPC. In some embodiments, the liposome comprises an OxPAPC that 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 a five-carbon sn-2 residue bearing omega-aldehyde or omega-carboxyl groups. In some embodiments, the liposome comprises an OxPAPC selected from HOdiA-PC, KOdiA-PC, HOOA-PC and KOOA-PC, or the OxPAPC is an epoxyisoprostane-containing phospholipid. In some embodiments, the liposome comprises an OxPAPC selected from 1-palmitoyl-2-(5,6-epoxyisoprostane E2)-sn-glycero-3-phosphocholine (5,6 PEIPC), 1-palmitoyl-2-(epoxycyclopenten-one)-sn-glycero-3-phosphorylcholine (PECPC),1-palmitoyl-2-(epoxy-isoprostane E2)-sn-glycero-4-phosphocholine (PEIPC), 1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC); 1-palmitoyl-2-(9′oxo-nonanoyl)-sn-glycer-o-3-phosphocholine; 1-palmitoyl-2-arachinodoyl-sn-glycero-3-phospho-choline; 1-palmitoyl-2-myristoyl-sn-glycero-3-phospho-choline; 1-palmitoyl-2-hexadecyl-sn-glycero-3-phosphocholine; 1-palmitoyl-2-azelaoyl-sn-glycero-3-phosphocholine; and 1-palmitoyl-2-acetoyl-sn-glycero-3-phosphocholine. In some embodiments, the liposome comprises PGPC. In some embodiments, the OxPAPC within the liposome lipid bilayer is 0%-100% of total lipids, or any range therein between. 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 a PEG and the exterior of the liposome, optionally wherein the targeting moiety is attached to one or both of the PEG and the exterior of the liposome by a covalent bond. 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 liposome contains 1 to 1000, 50 to 750, 100 to 500, or 30 to 200 targeting moieties, or any range therein between. In some embodiments, the liposome further comprises an immunostimulating agent (such as 1,6-beta 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 additional embodiments, the liposome is anionic or neutral. In some embodiments, the liposome has a zeta potential that is less than or equal to zero. In some embodiments, the liposome has a zeta potential that 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 therein between. In other embodiments, the liposome is cationic. In some embodiments, the liposomal composition comprises a liposome that has a zeta potential that is more than zero. In some embodiments, the liposome has a zeta potential that is 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 therein between.
Trans-Crocetin Conjugating/Complexing agents
In some embodiments, the pharmaceutical composition administered according to the provided methods comprises:
In some embodiments, the pharmaceutical composition administered according to the provided methods is a fixed dose of conjugated/complexed trans-crocetin. In some embodiments, the administered conjugated/complexed trans-crocetin composition does not contain a fixed dose of liposomal trans-crocetin.
In some embodiments, Q is a monovalent counterion (e.g., a monovalent metal cation or a monovalent organic cation). In further embodiments, the monovalent counterion is selected from NH4+, Na+, Li+, K+, or a monovalent organic cation such as protonated amine. In particular embodiments, the monovalent counterion is Na+.
In particular embodiments, the composition comprises sodium trans-crocetinate (STC).
In some embodiments, Q is a multivalent counterion (e.g., a multivalent cation such as a divalent metal cation or a divalent organic cation). In further embodiments, the multivalent cation is a divalent cation selected from Ca2+, Mg2+, Zn2+, Cu2+, Co2+, and Fe2+, a divalent organic cation such as protonated diamine, or a trivalent cation such as Fe3.
In some embodiments, the pharmaceutical composition administered according to the provided methods comprises:
In further embodiments, the conjugating/complexing agent is cyclodextrin. There are no particular limitations on the cyclodextrin contained in the provided pharmaceutical compositions so long as the cyclodextrins can complex trans-crocetin.
In particular embodiments, the cyclodextrin of the pharmaceutical composition is underivatized.
In other particular embodiments, the cyclodextrin is derivatized to bear ionizable (e.g., weakly basic and/or weakly acidic) functional groups to facilitate complexation with trans-crocetin.
Modifications of the hydroxyl groups of cyclodextrins, such as those facing away from the cyclodextrin interior phase, with ionizable chemical groups is known to facilitate the loading of cyclodextrins and therapeutic agents complexed with the cyclodextrins. In some embodiments, the cyclodextrins in the pharmaceutical composition have at least 2, 3, 4, 5, 6, 6, 7, 8, 9, or 10 hydroxyl group substituted with an ionizable chemical group. The term “charged cyclodextrin” refers to a cyclodextrin having one or more of its hydroxyl groups substituted with a charged moiety. Such a moiety can itself be a charged group or it can comprise an organic moiety (e.g., a C1-C6 alkyl or C1-C6 alkyl ether moiety) substituted with one or more charged moieties.
In some embodiments, the “ionizable” or “charged” moieties of a cyclodextrin derivative in the pharmaceutical compositions are weakly ionizable. Weakly ionizable moieties are those that are either weakly basic or weakly acidic. Weakly basic functional groups (W) have a pKa of between about 6.0-9.0, 6.5-8.5, 7.0-8.0, 7.5-8.0, and any range in between inclusive according to CH3-W. Similarly, weakly acidic functional groups (X) have a log dissociation constant (pKa) of between about 3.0-7.0, 4.0-6.5, 4.5-6.5, 5.0-6.0, 5.0-5.5, and any range in between inclusive according to CH3-X. Representative anionic moieties include, without limitation, carboxylate, carboxymethyl, succinyl, sulfonyl, phosphate, sulfoalkyl ether, sulphate carbonate, thiocarbonate, dithiocarbonate, phosphate, phosphonate, sulfonate, nitrate, and borate groups. Representative cationic moieties include, without limitation, amino, guanidine, and quaternary ammonium groups.
In another embodiment, the pharmaceutical composition comprises a derivatized cyclodextrin that is a “polyanion” or “polycation.” A polyanion is a derivatized cyclodextrin having more than one negatively charged group resulting in net a negative ionic charge of more than two units. A polycation is a derivatized cyclodextrin having more than one positively charged group resulting in net positive ionic charger of more than two units.
In another embodiment, the pharmaceutical composition comprises a derivatized cyclodextrin that is a “chargeable amphiphile.” By “chargeable” is meant that the amphiphile has a pK pH 4 to pH 8 or 8.5. A chargeable amphiphile may therefore be a weak acid or base. By “amphoteric” herein is meant a derivatized cyclodextrin having a ionizable groups of both anionic and cationic character wherein: (a) at least one, and optionally both, of the cation and anionic amphiphiles is chargeable, having at least one charged group with a pK between 4 and 8 to 8.5, (b) the cationic charge prevails at pH 4, and (c) the anionic charge prevails at pH 8 to 8.5.
In some embodiments, the “ionizable” or “charged” derivatized cyclodextrin as a whole, whether polyionic, amphiphilic, or otherwise, are weakly ionizable (i.e., have a pKai of between about 4.0-8.5, 4.5-8.0, 5.0-7.5, 5.5-7.0, 6.0-6.5, and any range in between inclusive).
Any one, some, or all hydroxyl groups of any one, some or all α-D-glucopyranoside units of a cyclodextrin can be modified to an ionizable chemical group as provided herein. Since each cyclodextrin hydroxyl group differs in chemical reactivity, reaction with a modifying moiety can produce an amorphous mixture of positional and optical isomers. Alternatively, certain chemistry can allow for pre-modified α-D-glucopyranoside units to be reacted to form uniform products.
The aggregate substitution that occurs for cyclodextrin derivatives in a mixture is described by a term referred to as the degree of substitution. For example, a 6-ethylenediamino-β-cyclodextrin with a degree of substitution of seven would be composed of a distribution of isomers of 6-ethylenediamino-β-cyclodextrin in which the average number of ethylenediamino groups per 6-ethylenediamino-β-cyclodextrin molecule is seven. The degree of substitution for a cyclodextrin derivative mixture can routinely be determined using mass spectrometry or nuclear magnetic resonance spectroscopy.
In one embodiment, at least one hydroxyl moiety facing away from the cyclodextrin interior is substituted with an ionizable chemical group. For example, the C2, C3, C6, C2 and C3, C2 and C6, C3 and C6, and all three of C2-C3-C6 hydroxyls of at least one α-D-glucopyranoside unit are substituted with an ionizable chemical group. Any such combination of hydroxyls can similarly be combined with at least two, three, four, five, six, seven, eight, nine, ten, eleven, up to all of the alpha-D-glucopyranoside units in the modified cyclodextrin as well as in combination with any degree of substitution provided herein. One such derivative is a sulfoalkyl ether cyclodextrin (SAE-CD). Sulfobutyl ether derivatives of beta cyclodextrin (SBE-β-CD) have been demonstrated to have significantly improved aqueous solubility compared to the parent cyclodextrin.
Additional cyclodextrin derivatives that may be complexed with trans-crocetin in the provided pharmaceutical compositions include sugammadex or Org to 25969, in which the 6-hydroxy groups on γ-CD have been replaced by carboxythio acetate ether linkages, and hydroxybutenyl-β-CD. Alternative forms of cyclodextrin include: 2,6-Di-O-methyl-β-CD (DIMEB), 2-hydroxylpropyl-3-cyclodextrin (HP-β-CD), randomly methylated-β-cyclodextrin (RAMEB), sulfobutyl ether ρ-cyclodextrin (SBE-β-CD), and sulfobutyl-ether-γ-cyclodextrin (SBEγCD), sulfobutylated beta-cyclodextrin sodium salt, sulfobutylated beta-cyclodextrin sodium salt, (2-Hydroxypropyl)-alpha-cyclodextrin, (2-Hydroxypropyl)-beta-cyclodextrin, (2-Hydroxypropyl)-γ-cyclodextrin, 2,6-di-O-methyl)-beta-cyclodextrin (DIMEB-50 Heptakis), 2,3,6-tri-O-methyl)-beta-cyclodextrin (TRIMEB Heptakis), methyl-beta-cyclodextrin, octakis (6-deoxy-6-iodo)-γ-cyclodextrin, and, octakis (6-deoxy-6-bromo)-gamma-cyclodextrin.
In some embodiments, a large association constant between the cyclodextrin and the trans-crocetin is preferable and can be obtained by selecting the number of glucose units in the cyclodextrin based on the size of therapeutic agent (see, for example, Albers et al., Crit. Rev. Therap. Drug Carrier Syst. 12:311-337 (1995); Stella et al., Toxicol. Pathol. 36:30-42 (2008). When the association constant depends on pH, the cyclodextrin can be selected such that the association constant becomes large at the pH of the composition. As a result, the solubility (nominal solubility) of the liposomal trans-crocetin in the presence of cyclodextrin can be further improved. In some embodiments, the association constant of the cyclodextrin with the trans-crocetin is 100, 200, 300, 400, 500, 600, 700, 800, 900, 1,000, or higher. In some embodiments, the association constant of the cyclodextrin with the trans-crocetin is 100-1,200, 200-1,000, 300-750, and any range in between inclusive.
In some embodiments, the cyclodextrin derivative of the pharmaceutical composition has the structure of Formula I:
In some embodiments, the cyclodextrin derivative of the pharmaceutical composition has the structure of formula II:
In some embodiments, the pharmaceutical composition comprises a cyclodextrin derivative disclosed in U.S. Pat. Nos. 6,133,248, 5,874,418, 6,046,177, 5,376,645, 5,134,127, 7,034,013, 6,869,939; and Intl. Publ. No. WO 02005/117911, the contents each of which is herein incorporated by reference in its priority.
In some embodiments, the pharmaceutical composition comprises sulfoalkyl ether cyclodextrin. In some embodiments, the cyclodextrin derivative is a sulfobutyl ether-3-cyclodextrin such as CAPTISOL® (CyDex Pharma Inc., Lenexa, Kans.). Methods for preparing sulfobutyl ether-3-cyclodextrin and other sulfoalkyl ether cyclodextrins are known in the art.
In some embodiments, the pharmaceutical composition comprises cyclodextrin derivative in the pharmaceutical composition is a compound of Formula III:
In some embodiments, the pharmaceutical composition comprises α-cyclodextrin, β-cyclodextrin, 2-hydroxypropyl-β-cyclodextrin, and/or 2-hydroxypropyl-γ-cyclodextrin, or γ-cyclodextrin. In particular embodiments, the pharmaceutical composition comprises γ-cyclodextrin.
In some embodiments, the molar ratio of trans-crocetin/cyclodextrin is 1: 1-20, or any range therein between (e.g., 1:1-10, 1:2-8, 1:1-5, 1:3-5, 1:3, 1:4, or 1:5). In some embodiments, the molar ratio trans-crocetin/cyclodextrin is: 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, or 1:15, or 1:>15.
In some embodiments, the pharmaceutical composition comprises γ-cyclodextrin and the molar ratio of trans-crocetin/γ-cyclodextrin is 1: 1-20, or any range therein between (e.g., 1:1-10, 1:2-8, 1:1-5, 1:3-5, 1:3, 1:4, or 1:5). In some embodiments, the molar ratio trans-crocetin/γ-cyclodextrin is: 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8,1:9, 1:10, or 1:15, or >15:1.
In some embodiments, the molar ratio of trans-crocetin/cyclodextrin in the pharmaceutical composition is 1-20:1, or any range therein between (e.g., 1-10:1, 2-8:1, 1-5:1, 3:1, 4:1, or 5:1). In some embodiments, the molar ratio of trans-crocetin/cyclodextrin in the pharmaceutical composition is: is: 3:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, or 15:1, or >15:1.
In some embodiments, the cyclodextrin concentration in the pharmaceutical composition is 1-15%, or any range therein between (e.g., 5-10%). In some embodiments, the cyclodextrin concentration in the pharmaceutical composition is 4%, 5%, 6%, 7%, 8%, 9%, or 10%.
In some embodiments, the pH of the pharmaceutical composition 6-10, 7.5-9.5, or 8-9 (e.g., pH 8.5), or any range therein between. In some embodiments, the pharmaceutical composition comprises a buffer having a pKA within 1 unit or within 0.5 units of the pH of the solution at a concentration of 1-200 mM, 1-100 mM, 1-80 mM, or any range therein between. In further embodiments, the pharmaceutical composition comprises a buffer selected from: glycine, gly-gly, sodium bicarbonate, sodium phosphate, tricine, bicine, EPPS (HEPPS), HEPBS, TABS, AMPD, or sodium borate (e.g., glycine, gly-gly, or sodium bicarbonate. In particular embodiments, the pharmaceutical composition comprises glycine or sodium bicarbonate.
In some embodiments, the pharmaceutical composition comprises one or more tonicity agents. In embodiments, the tonicity agent is dextrose, mannitol, glycerin, potassium chloride, or sodium chloride. In some embodiments the pharmaceutical composition comprises a tonicity agent at a concentration of greater than 0.1%, or a concentration of 0.3% to 2.5%, or any range therein between. In some embodiments, the pharmaceutical composition comprises dextrose, mannitol, glycerin, potassium chloride, or sodium chloride at a concentration of greater than 0.1%, or a concentration of 0.3% to 2.5%, or any range therein between. In some embodiments, the pharmaceutical composition comprises trehalose or dextrose. In some embodiments, the pharmaceutical composition comprises mannitol.
The provided compositions can be formulated in whole or in part as pharmaceutical compositions. Pharmaceutical compositions may include one or more nanoparticle compositions. For example, a pharmaceutical composition may include one or more nanoparticle compositions including one or more different therapeutic and/or prophylactics. Pharmaceutical compositions may further include one or more pharmaceutically acceptable excipients or accessory ingredients such as those described herein. General guidelines for the formulation and manufacture of pharmaceutical compositions and agents are available, for example, in Remington's The Science and Practice of Pharmacy, 21′ Edition, A. R. Gennaro; Lippincott, Williams & Wilkins, Baltimore, Md., 2006. Conventional excipients and accessory ingredients may be used in any pharmaceutical composition, except insofar as any conventional excipient or accessory ingredient may be incompatible with one or more components of a nanoparticle composition. An excipient or accessory ingredient may be incompatible with a component of a nanoparticle composition if its combination with the component may result in any undesirable biological effect or otherwise deleterious effect.
In some embodiments, one or more excipients or accessory ingredients may make up greater than 50% of the total mass or volume of a pharmaceutical composition including a nanoparticle composition. For example, the one or more excipients or accessory ingredients may make up 50%, 60%, 70%, 80%, 90%, or more of a pharmaceutical convention. In some embodiments, a 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, an excipient is approved for use in humans and for veterinary use. In some embodiments, an excipient is approved by United States Food and Drug Administration. In some embodiments, an excipient is pharmaceutical grade. In some embodiments, an excipient meets the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia. and/or the International Pharmacopoeia.
Standard methods for making liposomes include, but are not limited to, methods reported in Liposomes: A Practical Approach, V. P. Torchilin, Volkmar Weissig Oxford University Press, 2003 and are known in the art.
In some embodiments, the disclosure provides a pharmaceutical composition and a physiologically (i.e., pharmaceutically) acceptable carrier. As used herein, the term “carrier” refers to a typically inert substance used as a diluent or vehicle for a drug such as a therapeutic agent. The term also encompasses a typically inert substance that imparts cohesive qualities to the composition. Typically, the physiologically acceptable carriers are present in liquid form. Examples of liquid carriers include physiological saline, phosphate buffer, normal buffered saline (135-150 mM NaCl), water, buffered water, 0.4% saline, 0.3% glycine, glycoproteins to provide enhanced stability (e.g., albumin, lipoprotein, globulin, etc.), and the like. Since physiologically acceptable carriers are determined in part by the particular composition being administered as well as by the particular method used to administer the composition, there are a wide variety of suitable formulations of pharmaceutical compositions provided herein (See, e.g., Remington's Pharmaceutical Sciences, 17th ed., 1989).
The provided compositions may be sterilized by conventional, known sterilization techniques or may be produced under sterile conditions. Aqueous solutions can be packaged for use or filtered under aseptic conditions and lyophilized, the lyophilized preparation being combined with a sterile aqueous solution prior to administration. The compositions can contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, tonicity adjusting agents, wetting agents, and the like, e.g., sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, sorbitan monolaurate, and triethanolamine oleate. Sugars can also be included for stabilizing the compositions, such as a stabilizer for lyophilized trans-crocetin compositions. In some embodiments, the pharmaceutical composition comprises a tonicity agent at a concentration of 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 therein between. In some embodiments, the pharmaceutical composition comprises a tonicity agent such as dextrose, mannitol, glycerin, potassium chloride, or sodium chloride. In further embodiments, the pharmaceutical composition comprises dextrose, mannitol, glycerin, potassium chloride, or sodium chloride at a concentration of 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 therein between.
Formulations suitable for parenteral administration, such as, for example, by intraarticular (in the joints), intravenous, intramuscular, intratumoral, intradermal, intraperitoneal, and subcutaneous routes, include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. Injection solutions and suspensions can also be prepared from sterile powders, granules, and tablets. In some embodiments, the provided pharmaceutical compositions are administered, for example, by intravenous infusion, topically, intraperitoneally, intravesically, or intrathecally. In particular embodiments, the pharmaceutical compositions are parentally or intravenously administered. Preferably, the pharmaceutical compositions are administered parentally, i.e. intraarticularly, intravenously, subcutaneously, or intramuscularly. In other embodiments, the pharmaceutical preparation may be administered topically.
In some embodiments, the fixed dose liposomal trans-crocetin composition is administered as an intravenous infusion once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, the fixed dose liposomal trans-crocetin composition is administered as an intravenous infusion two times a day (e.g., every 12 hours, +/−3 hours), for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, the fixed dose liposomal trans-crocetin composition is administered as an intravenous infusion once a day (e.g., every 24 hours, +/−6 hours), for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In particular embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 100 mg to 300 mg, 80 mg to 275 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg, or 225 mg to 275 mg (e.g., 250 mg), or any range therein between is administered as an intravenous infusion. In some embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of fixed dose of 100 mg-400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg), or any range therein between, is administered as an intravenous infusion. In some embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 200 mg to 300 mg (e.g., 250 mg), or any range therein between, is administered as an intravenous infusion. In some embodiments, the one or more liposomal trans-crocetin composition is administered as an intravenous infusion over 15 minutes to 5 hours, or any range therein between. In some embodiments, the one or more liposomal trans-crocetin composition is administered as an intravenous infusion over 2 hours to 4 hours, or any range therein between. In particular embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of fixed dose of 100 mg-400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg), or any range therein between, is administered as an intravenous infusion over 2 hours to 4 hours, or any range therein between. In other particular embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 200 mg to 300 mg (e.g., 250 mg), or any range therein between, is administered as an intravenous infusion over 2 hours to 4 hours, or any range therein between. In some embodiments, one or more fixed dose liposomal trans-crocetin compositions is administered as an intravenous infusion over 3 hours. In particular embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of fixed dose of 100 mg to 400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg), or any range therein between, is administered as an intravenous infusion over 3 hours. In other particular embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 200 mg to 300 mg (e.g., 250 mg), or any range therein between, is administered as an intravenous infusion over 3 hours. In some embodiments, the fixed dose liposomal trans-crocetin composition is administered as an intravenous infusion once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, the fixed dose liposomal trans-crocetin composition is administered as an intravenous infusion two times a day (e.g., every 12 hours, +/−3 hours), for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, the fixed dose liposomal trans-crocetin composition is administered as an intravenous infusion once a day (e.g., every 24 hours, +/−6 hours), for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In particular embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 25 mg to 900 mg, 60 mg to 600 mg, 150 mg to 600 mg (e.g., 550-600, 560 mg, or 580 mg), 70 mg to 580 mg, 150 mg to 550 mg (e.g., 200 mg to 400 mg, 250 mg to 350 mg, 250 mg, 300 mg, or 380 mg), 80 mg to 350 mg, 75 mg to 260 mg, or 25 mg to 250 mg (e.g., 120 mg to 160 mg or 140 mg), or any range therein between is administered as an intravenous infusion. In some embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 550 mg to 600 mg (e.g., 560,g or 580 mg), or any range therein between, is administered as an intravenous infusion. In some embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 100 mg to 400 mg (e.g., 100 mg to 200 mg, 200 mg to 300 mg, 200 mg to 300 mg, 140 mg, 300 mg, or 380 mg), or any range therein between, is administered as an intravenous infusion. In some embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 300 mg to 450 mg (e.g., 380 mg), or any range therein between, is administered as an intravenous infusion. In some embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 250 mg to 350 mg (e.g., 300 mg), or any range therein between, is administered as an intravenous infusion. In some embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 100 mg to 200 mg (e.g., 140 mg), or any range therein between, is administered as an intravenous infusion. In some embodiments, the one or more liposomal trans-crocetin composition is administered as an intravenous infusion over 15 minutes to 5 hours (e.g., 1 hour to 3 hours, 2 hours to 4 hours, 2 hours or 3 hours), or any range therein between. In particular embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of 250 mg to 350 mg (e.g., 300 mg), or any range therein between, is administered as an intravenous infusion over 15 minutes to 5 hours (e.g., 1 hour to 3 hours, 2 hours to 4 hours, 2 hours or 3 hours), or any range therein between. In particular embodiments, one or more liposomal trans-crocetin compositions at a fixed dosage of fixed dose of 100 mg-400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg), or any range therein between, is administered as an intravenous infusion over 15 minutes to 5 hours (e.g., 1 hour to 3 hours, 2 hours to 4 hours, 2 hours or 3 hours), or any range therein between. In some embodiments, the fixed dose liposomal trans-crocetin composition is administered to a subject as an intravenous infusion once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, the fixed dose liposomal trans-crocetin composition is administered to a subject as an intravenous infusion two times a day (e.g., every 12 hours, +/−3 hours), for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, the fixed dose liposomal trans-crocetin composition is administered to a subject as an intravenous infusion once a day (e.g., every 24 hours, +/−6 hours), for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In particular embodiments, one or more doses of liposomal trans-crocetin is administered to a subject at a fixed dosage of 250 mg to 350 mg (e.g., 300 mg), as an intravenous infusion over 15 minutes to 5 hours (e.g., 1 hour to 3 hours, 2 hours to 4 hours, 2 hours or 3 hours), four times daily, three times daily, 2 times daily, or once a day, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more days.
In particular embodiments, one or more doses of liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg), or any range therein between, as an intravenous infusion over 1 hour to 4 hours, four times daily, three times daily, 2 times daily, or once a day, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more days.
In particular embodiments, one or more doses of liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg), or any range therein between, as an intravenous infusion over 1 hour to 4 hours, four times daily, three times daily, 2 times daily, or once a day, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more days.
In particular embodiments, one or more doses of liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg)), or any range therein between, as an intravenous infusion over 1 hour to 3 hours, 2 times daily for 1, 2, 3, 4, 5, or more days. In particular embodiments, one or more doses of liposomal trans-crocetin is administered to a subject at a fixed dosage of 200 mg to 800 mg (e.g., 200 mg to 600 mg, 200 mg to 400 mg, 240 mg to 320 mg, 600 mg, or 280 mg), or any range therein between, as an intravenous infusion over 1 hour to 3 hours, once over 24 hours (+/−6 hours), for 1, 2, 3, 4, 5, or more days.
In particular embodiments, one or more doses of liposomal trans-crocetin is administered to a subject at a fixed dosage of 250 mg to 350 mg (e.g., 300 mg), or any range therein between, as an intravenous infusion over 1 hour to 3 hours, 2 times daily for 1, 2, 3, 4, 5, or more days. In particular embodiments, one or more doses of liposomal trans-crocetin is administered to a subject at a fixed dosage of 500 mg to 700 mg (e.g., 600 mg), or any range therein between, as an intravenous infusion over 1 hour to 3 hours, once a day (i.e., 24 hours (+/−6 hours)), for 1, 2, 3, 4, 5, or more days.
In some embodiments, one or more liposomal trans-crocetin compositions is administered as an intravenous infusion. In some embodiments, the one or more liposomal trans-crocetin composition is administered as an intravenous infusion over 15 minutes to 5 hours, or any range therein between. In some embodiments, the one or more liposomal trans-crocetin composition is administered as an intravenous infusion over 2 hours to 4 hours, or any range therein between. In some embodiments, the one or more liposomal trans-crocetin composition is administered as an intravenous infusion over 3 hours.
In some embodiments, the provided pharmaceutical compositions (e.g., liposomal compositions) are presented in unit-dose or multi-dose sealed containers, such as ampoules and vials.
In some embodiments, the pharmaceutical preparations are administered in unit dosage form. In such form the preparation is subdivided into unit doses containing appropriate quantities of the active component, e.g., a trans-crocetin composition. The unit dosage form can be a packaged preparation, the package containing discrete quantities of preparation. The composition can, if desired, also contain other compatible therapeutic agents (e.g., as described herein).
In some embodiments, the pharmaceutical compositions provided herein can be administered at one or more doses of 100 mg to 900 mg, 100 mg to 800 mg, 100 mg to 700 mg 100 mg to 600 mg (e.g., 550-600, 560 mg, or 580 mg), 100 mg to 500 mg, 100 mg to 400 mg (e.g., 200 mg to 400 mg, 250 mg to 350 mg, 300 mg to 400 mg, 250 mg, 300 mg, 350 mg, or 380 mg), or 100 mg to 300 mg (e.g., 120 mg to 160 mg or 140 mg), or any range therein between, of trans-crocetin (e.g., liposomal trans-crocetin).
In some embodiments, the pharmaceutical compositions provided herein can be administered at one or more doses of 100 mg to 300 mg, 80 mg to 275 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg, or 225 mg to 275 mg (e.g., 250 mg), or any range therein between, of trans-crocetin (e.g., liposomal trans-crocetin). In some embodiments, a plurality of pharmaceutical compositions are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, a plurality of pharmaceutical compositions are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In some embodiments, the trans-crocetin pharmaceutical compositions provided herein are administered at one or more doses of 250 mg and/or one or more doses of 140 mg of trans-crocetin (e.g., liposomal trans-crocetin). In some embodiments, two or more trans-crocetin pharmaceutical compositions are administered to a subject mg 1 hour to 48 hours, 1.5 hours to 24 hours, 2 hours to 18 hours, 4 hours to 16 hours (e.g., 12 hours (+/−3 hours)), or 1 hour to 8 hours (e.g., 3 hours) apart, or any range therein between. In some embodiments, a plurality of pharmaceutical compositions are administered to a subject four times a day, three times a day, twice a day, once a day, or once every other day.
In a particular embodiment, on day 1 of a dosing regimen/treatment method provided herein, two fixed loading doses of 200-300 mg liposomal trans-crocetin, or any range therein between are administered 12 hours apart (+/−3 hours) as an IV infusion over 1-3 hours (e.g., 2 hours). And on Day 2 onwards, two fixed maintenance doses of 100-200 mg liposomal trans-crocetin, or any range therein between liposomal trans-crocetin are administered once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In a particular embodiment, on day 1 of a dosing regimen/treatment method provided herein, two fixed loading doses of 250 mg liposomal trans-crocetin are administered 12 hours apart (+/−3 hours) as an IV infusion over 1-3 hours (e.g., 2 hours). And on Day 2 onwards, two fixed maintenance doses of 140 mg liposomal trans-crocetin are administered once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In some embodiments, the trans-crocetin pharmaceutical compositions provided herein are administered at the initial total daily dose of 100 mg to 920 mg, 400 mg-650 mg, 600 mg to 900 mg (e.g., 600 mg), 200 mg to 400 mg (e.g., 280 mg), or 300 mg to 400 mg over a period of 24 hours (+/−6 hours). In some embodiments, two or more trans-crocetin pharmaceutical compositions are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, a plurality of pharmaceutical compositions are administered to a subject four times a day, three times a day, twice a day, once a day, or once every other day.
In a particular embodiment, on day 1 of a dosing regimen/treatment method provided herein, two fixed loading doses of 100 mg to 460 mg, 100 mg-350 mg, 100 mg to 200 mg or 150 mg to 200 mg, or any range therein between, of liposomal trans-crocetin, or any range therein between are administered 12 hours apart (+/−3 hours) as an IV infusion over 1-3 hours (e.g., 2 hours). And on Day 2 onwards, one or more fixed maintenance dose of 200 mg to 700 mg, 300 mg to 400 mg or 200 mg to 400 mg, liposomal trans-crocetin, or any range therein between is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In a particular embodiment, on day 1 of a dosing regimen/treatment method provided herein, two loading doses of 100 mg to 460 mg, or any range therein between of liposomal trans-crocetin, are administered 12 hours apart (+/−3 hours) as an IV infusion over 1-3 hours (e.g., 2 hours). And on Day 2 onwards, one maintenance dose of 200 mg to 600 mg, or any range therein between, of liposomal trans-crocetin, is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
The dosages, however, may be varied depending upon the requirements of the patient, the severity of the condition being treated, and the trans-crocetin composition being employed. For example, dosages can be empirically determined considering the type and stage of the disorder or condition diagnosed in a particular patient. The dose administered to a patient, in the context of the provided pharmaceutical compositions (e.g., a liposomal trans-crocetin composition) should be sufficient to affect a beneficial therapeutic response in the patient over time. For convenience, the total daily dosage may be divided and administered in portions during the day, if desired.
In one particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In one particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In another particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg) once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In another particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg) once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In a further particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg), or any range therein between followed by the administration of liposomal trans-crocetin at a fixed dose of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a further particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg), or any range therein between, for 1 day (day 1), followed by the administration of liposomal trans-crocetin at a fixed dose of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In yet a further particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg), or any range therein between, once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1 day (day 1), followed by the administration of liposomal trans-crocetin at a fixed dose of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In another particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 300 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a further particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dose of 300 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month followed by the administration of liposomal trans-crocetin at a fixed dose of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a further particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dose of 300 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1 day (day 1), followed by the administration of liposomal trans-crocetin at a fixed dose of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1-20, 1-15, 1-10 or 1-5 days. In yet a further particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dose of 300 mg for 1 day (day 1), followed by the administration of liposomal trans-crocetin at a fixed dose of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In particular embodiments, the liposomal trans-crocetin is administered to a subject (e.g., a human) experiencing acute lung distress (e.g., presenting symptoms such as having difficulty breathing, tachypnea, mental confusion due to low oxygen levels) and/or having a PaO2/FiO2 ratio of less than 300 mm Hg. In other particular embodiments, the liposomal trans-crocetin is administered to a subject (e.g., a human) experiencing Acute Respiratory ARDS and/or having a PaO2/FiO2 ratio of less than 200 mm Hg. In other particular embodiments, the liposomal trans-crocetin is administered to a subject in order to increase the patients PaO2/FiO2 ratio. In some embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 5%-75%, or any range therein between. In further embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 5%, 10%, 15%, 20%, 25%, 30% 40% or 50%. In particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 10%. In other particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 25%. In other particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 40%.
Animal toxicology studies have indicated efficacy without dose limiting toxicity at liposomal trans-crocetin doses as high as 25 mg/kg. As disclosed herein, dose limiting toxicity of liposomal trans-crocetin has not been observed in humans and liposomal trans-crocetin has been administered at doses as high as 7.5 mg/kg in humans. In particular embodiments, the administered composition comprises liposomal trans-crocetin and is administered to a subject (e.g., human) at a fixed dosage of about 100 mg to 900 mg, 100 mg to 800 mg, 100 mg to 700 mg 100 mg to 600 mg (e.g., 550-600, 560 mg, or 580 mg), 100 mg to 500 mg, 100 mg to 400 mg (e.g., 200 mg to 400 mg, 250 mg to 350 mg, 300 mg to 400 mg, 250 mg, 300 mg, 350 mg, or 380 mg), 100 mg to 300 mg, or 100 mg to 200 mg (e.g., 120 mg to 160 mg or 140 mg), or any range therein between. In some embodiments, fixed dose of liposomal trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a particular embodiment, liposomal trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a particular embodiment, liposomal trans-crocetin is administered to a subject once over a 24 hour period (+/−6 hours)).
As disclosed herein, the inventors have surprisingly found that the pharmacokinetic characteristics of trans-crocetin highlight its safety and efficacy across different dosing regimens and show stable levels of AUC and Cmax across a wide range of body weights. The pharmacokinetic profile of trans-crocetin substantiates a fixed dosing strategy for trans-crocetin from a pharmacokinetic perspective. In one embodiment, a fixed dose of liposomal trans-crocetin administered to a subject (e.g., human) at a dosage of about 5 mg to 900 mg, 60 mg to 600 mg (e.g., 580 mg), 150 mg to 600 mg, 70 mg to 580 mg, 150 mg to 550 mg, 80 mg to 350 mg, 75 mg to 260 mg, 25 mg to 250 mg, 200 mg to 400 mg, 300 mg to 400 mg, or any range therein between. In another embodiment, a fixed dose of liposomal trans-crocetin administered to a subject (e.g., human) at a dosage of about 100 mg to 300 mg, 80 mg to 275 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg (e.g., 300 mg), or 225 mg to 275 mg (e.g., 250 mg), or any range therein between. In some embodiments, one or more fixed doses of liposomal trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a particular embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In one embodiment, a fixed dose of liposomal trans-crocetin administered to a subject (e.g., human) at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg). In one embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) at a dosage of about 140 mg. In a further embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg), or any range therein between, and a fixed dose of liposomal trans-crocetin is administered to the subject at a dosage of about 140 mg. In some embodiments, one or more fixed doses of liposomal trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a particular embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In one embodiment, a fixed dose of liposomal trans-crocetin administered to a subject (e.g., human) at a dosage of about 300 mg. In one embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) at a dosage of about 140 mg. In a further embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) at a dosage of about 300 mg and a fixed dose of liposomal trans-crocetin is administered to the subject at a dosage of about 140 mg. In some embodiments, one or more fixed doses of liposomal trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a particular embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In one embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month at a dosage of about 100 mg to 300 mg, 80 mg to 275 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg, or 225 mg to 275 mg (e.g., 250 mg), or any range therein between. In one embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month at a dosage of about 100 mg to 300 mg, 80 mg to 275 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg, or 225 mg to 275 mg (e.g., 250 mg), or any range therein between. In one embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month at a dosage of about 300 mg. In one embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg). In one embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month at a dosage of about 140 mg. In a further embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month at a dosage of about 300 mg and two or more fixed doses of liposomal trans-crocetin is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month at a dosage of about 140 mg. In a further embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject (e.g., human) once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg), or any range therein between, and two or more fixed doses of liposomal trans-crocetin is administered to the subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month at a dosage of about 140 mg.
In some embodiments, the fixed dose of liposomal trans-crocetin administered to a patient depends on the type of disorder or condition to be treated disease to be treated, the severity and course of the disease, whether the trans-crocetin is administered for preventive or therapeutic purposes, previous therapy, the patient's clinical history and response to the trans-crocetin, and the discretion of the attending physician. The fixed dose is suitably administered to the patient at one time or over a series of treatments.
In particular embodiments, liposomal trans-crocetin is administered to a subject (e.g., a human) experiencing acute lung distress (e.g., presenting symptoms such as having difficulty breathing, tachypnea, mental confusion (e.g., due to low oxygen levels)) and/or having a PaO2/FiO2 ratio of less than 300 mm Hg or less than 250 mm Hg. In other particular embodiments, the trans-crocetin is administered to a subject (e.g., a human) experiencing ARDS and/or having a PaO2/FiO2 ratio of less than 200 mm Hg. In other particular embodiments, trans-crocetin is administered to a subject in order to increase the patients PaO2/FiO2 ratio. In some embodiments, the administration of trans-crocetin increases the patient's PaO2/FiO2 ratio by 5%-75%, or any range therein between. In some embodiments, administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 10%-50%, or any range therein between. In further embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 5%, 10%, 15%, 20%, 25%, 30% 40% or 50%.
In particular embodiments, liposomal trans-crocetin is administered to a subject (e.g., a human) experiencing acute lung distress (e.g., presenting symptoms such as having difficulty breathing, tachypnea, mental confusion (e.g., due to low oxygen levels)) and/or having a PaO2/FiO2 ratio of less than 300 mm Hg or less than 250 mm Hg. In other particular embodiments, liposomal trans-crocetin is administered to a subject (e.g., a human) experiencing ARDS and/or having a PaO2/FiO2 ratio of less than 200 mm Hg. In other particular embodiments, liposomal trans-crocetin is administered to a subject in order to increase the patients PaO2/FiO2 ratio. In some embodiments, the administration of liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 5%-75%, or any range therein between. In some embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 10%-50%, or any range therein between. In further embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 5%, 10%, 15%, 20%, 25%, 30% 40% or 50%.
In some embodiments, liposomal trans-crocetin is administered to a subject (e.g., human) at a fixed dosage of 100 mg to 900 mg, 100 mg to 800 mg, 100 mg to 700 mg 100 mg to 600 mg (e.g., 550-600, 560 mg, or 580 mg), 100 mg to 500 mg, 100 mg to 400 mg (e.g., 200 mg to 400 mg, 250 mg to 350 mg, 300 mg to 400 mg, 250 mg, 300 mg, 350 mg, or 380 mg), or 100 mg to 300 mg (e.g., 120 mg to 160 mg or 140 mg), or any range therein between, of liposomal trans-crocetin. In some embodiments the liposomal trans-crocetin is administered at a fixed dosage of 100 mg to 300 mg, 80 mg to 275 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg, or 225 mg to 275 mg (e.g., 250 mg), or any range therein between of liposomal trans-crocetin. In particular embodiments, the liposomal trans-crocetin is administered at a fixed dosage of 140 mg. In additional particular embodiments, the liposomal trans-crocetin is administered at a fixed dosage of 250 mg. In further particular embodiments, the liposomal trans-crocetin is administered at a fixed dosage of 250 mg and at a fixed dosage of 140 mg. In some embodiments, the fixed dose of liposomal trans-crocetin is administered once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a particular embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In one particular embodiment, a fixed dose of liposomal trans-crocetin is administered to a subject at a fixed dosage of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In another particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg) once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a further particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg), or any range therein between, once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month followed by the administration of liposomal trans-crocetin at a fixed dose of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a further particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg), or any range therein between, once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1 day (day 1), followed by the administration of liposomal trans-crocetin at a fixed dose of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1-20, 1-15, 1-10 or 1-5 days. In yet a further particular embodiment, liposomal trans-crocetin is administered to a subject at a fixed dosage of 100 mg to 350 mg (e.g., 250 mg or 300 mg), or any range therein between, once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1 day (day 1), followed by the administration of liposomal trans-crocetin at a fixed dose of 140 mg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 5%-75%, or any range therein between, after 4, 3, 2, or 1 trans-crocetin treatment. In particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 20% after 4, 3, 2, or 1 trans-crocetin treatment. In further particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 25% after 4, 3, 2, or 1 trans-crocetin treatment. In further particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 30% after 4, 3, 2, or 1 trans-crocetin treatment.
In some embodiments, liposomal trans-crocetin is administered at a dosage of 2.5 mg/kg to 7.5 mg/kg, or any range therein between, once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a particular embodiment, the liposomal trans-crocetin is administered once a day. In some embodiments, the liposomal trans-crocetin is administered once a week. In some embodiments, the liposomal trans-crocetin is administered once a month. In particular embodiments, liposomal trans-crocetin is administered at a dosage of 2.5 mg/kg to 7.5 mg/kg, or any range therein between, once daily. In particular embodiments, liposomal trans-crocetin is administered at a dosage of 2.5 mg/kg to 7.5 mg/kg, or any range therein between, once daily for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 5%-75%, or any range therein between, after 4, 3, 2, or 1 trans-crocetin treatment. In particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 20% after 4, 3, 2, or 1 trans-crocetin treatment. In further particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 25% after 4, 3, 2, or 1 trans-crocetin treatment. In further particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 30% after 4, 3, 2, or 1 trans-crocetin treatment.
In some embodiments, liposomal trans-crocetin is administered at a dosage of about 2.5 mg/kg, once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In a particular embodiment, the liposomal trans-crocetin is administered once a day. In some embodiments, the liposomal trans-crocetin is administered once a week. In some embodiments, the liposomal trans-crocetin is administered once a month. In particular embodiments, liposomal trans-crocetin is administered at a dosage of about 2.5 mg/kg once daily. In particular embodiments, liposomal trans-crocetin is administered at a dosage of about 2.5 mg/kg once daily for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 5%-75%, or any range therein between, after 4, 3, 2, or 1 trans-crocetin treatment. In particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 20% after 4, 3, 2, or 1 trans-crocetin treatment. In further particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 25% after 4, 3, 2, or 1 trans-crocetin treatment. In further particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 30% after 4, 3, 2, or 1 trans-crocetin treatment.
In particular embodiments, liposomal trans-crocetin is administered at a dosage of about 5 mg/kg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, the liposomal trans-crocetin is administered once a day. In some embodiments, the liposomal trans-crocetin is administered once a week. In some embodiments, the liposomal trans-crocetin is administered once a month. In particular embodiments, liposomal trans-crocetin is administered at a dosage of about 5 mg/kg once daily. In particular embodiments, liposomal trans-crocetin is administered at a dosage of about 5 mg/kg once daily for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 5%-75%, or any range therein between, after 4, 3, 2, or 1 trans-crocetin treatment. In particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 20% after 4, 3, 2, or 1 trans-crocetin treatment. In further particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 25% after 4, 3, 2, or 1 trans-crocetin treatment. In further particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 30% after 4, 3, 2, or 1 trans-crocetin treatment.
In particular embodiments, liposomal trans-crocetin is administered at a dosage of about 7.5 mg/kg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, the liposomal trans-crocetin is administered once a day. In some embodiments, the liposomal trans-crocetin is administered once a week. In some embodiments, the liposomal trans-crocetin is administered once a month. In particular embodiments, liposomal trans-crocetin is administered at a dosage of about 7.5 mg/kg once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, the liposomal trans-crocetin is administered once a day. In some embodiments, the liposomal trans-crocetin is administered once a week. In some embodiments, the liposomal trans-crocetin is administered once a month. In particular embodiments, liposomal trans-crocetin is administered at a dosage of about 5 mg/kg once daily for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In some embodiments, administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 5%-75%, or any range therein between, after 4, 3, 2, or 1 trans-crocetin treatment. In particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 20% after 4, 3, 2, or 1 trans-crocetin treatment. In further particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 25% after 4, 3, 2, or 1 trans-crocetin treatment. In further particular embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 30% after 4, 3, 2, or 1 trans-crocetin treatment.
The fixed dose of liposomal trans-crocetin is optimally administered to the patient at one time or over a series of treatment periods. In some embodiments, the fixed dose of liposomal trans-crocetin is about 50 mg to about 600 mg (e.g., 50 mg to 300 mg, 150 mg to 350 mg, or 150 to 550 mg). In some embodiments, the fixed dose of liposomal trans-crocetin is 80 mg to 140 mg, 140 mg to 220 mg, or 220 mg to 450 mg of trans-crocetin. In some embodiments, the fixed dose of liposomal trans-crocetin is 25 mg to 900 mg, 60 mg to 600 mg, 150 mg to 600 mg, 70 mg to 580 mg, 150 mg to 550 mg, 200 mg to 400 mg, 300 mg to 400 mg, 80 mg to 350 mg, 75 mg to 260 mg, or 25 mg to 250 mg, or any range therein between. In some embodiments, the fixed dose of liposomal trans-crocetin is 100 mg to 300 mg, 80 mg to 275 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg, or 225 mg to 275 mg (e.g., 250 mg), or any range therein between.
In particular embodiments, a fixed dose of liposomal trans-crocetin is optimally administered to the patient over a series of treatment periods. In some embodiments, the fixed dose of liposomal trans-crocetin is about 50 mg to about 600 mg (e.g., 50 mg to 300 mg, 150 mg to 350 mg, or 150 to 550 mg). In some embodiments, the fixed dose of liposomal trans-crocetin is 80 mg to 140 mg, 140 mg to 220 mg, or 220 mg to 450 mg, or any range therein between. In some embodiments, the fixed dose of liposomal trans-crocetin is 100 mg to 900 mg, 100 mg to 800 mg, 100 mg to 700 mg 100 mg to 600 mg (e.g., 550-600, 560 mg, or 580 mg), 100 mg to 500 mg, 100 mg to 400 mg (e.g., 200 mg to 400 mg, 250 mg to 350 mg, 300 mg to 400 mg, 250 mg, 300 mg, 350 mg, or 380 mg), or 100 mg to 300 mg (e.g., 120 mg to 160 mg or 140 mg), or any range therein between. In some embodiments, the fixed dose of liposomal trans-crocetin is 100 mg to 300 mg, 80 mg to 275 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg, or 225 mg to 275 mg (e.g., 250 mg), or any range therein between.
In one embodiment, one or more loading dose(s) of trans-crocetin is followed by a plurality of fixed maintenance doses of trans-crocetin. In another embodiment, one or more fixed loading dose(s) of trans-crocetin is followed by a plurality of fixed maintenance doses of trans-crocetin. In other embodiments, multiple identical fixed doses of trans-crocetin are administered to the patient. In one embodiment, a fixed dose (loading dose) of 100 mg to 550 mg (e.g., 100 mg to 300 mg, 150 mg to 550 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg), or any range therein between, of liposomal trans-crocetin is followed by a fixed dose of approximately 100 mg to 400 mg (e.g., 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg), (maintenance dose) of trans-crocetin. In particular embodiments, two fixed loading dose of trans-crocetin are administered within 24 hours. In some embodiments, a plurality of fixed maintenance doses of trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In particular embodiments, two fixed maintenance doses of trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In another embodiment, one or more loading dose(s) of liposomal trans-crocetin is followed by a plurality of fixed maintenance doses of liposomal trans-crocetin.
In one embodiment, one or more loading dose(s) of liposomal trans-crocetin is followed by a plurality of fixed maintenance doses of liposomal trans-crocetin. In other embodiments, multiple identical doses of liposomal trans-crocetin are administered to the patient. In one embodiment, a dose (loading dose) 200 mg to 920 mg (e.g., 908 mg, 300 mg, or 250 mg), or any range therein between, of liposomal trans-crocetin is followed by a plurality of doses (maintenance doses) 100 mg to 350 mg (e.g., 200 mg to 350 mg, or 300 mg), 100 mg to 200 mg or 150 mg to 200 mg of liposomal trans-crocetin, or any range therein between, of liposomal trans-crocetin. In some embodiments, two loading dose of 200 mg to 920 mg, or any range therein between, of liposomal trans-crocetin are administered to a subject within 24 hours of each other. In some embodiments, a plurality of fixed maintenance doses of 100 mg to 350 mg, or any range therein between, of liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In particular embodiments, a maintenance dose of liposomal trans-crocetin are administered to the subject once a day, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In another particular embodiments, a maintenance dose of liposomal trans-crocetin is administered to the subject once a week, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In another particular embodiments, a maintenance dose of liposomal trans-crocetin is administered to the subject once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In one embodiment, the subject is administered a fixed loading dose of 100 mg to 550 mg (e.g., 100 mg to 300 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg,380 mg, or 550 mg), or any range therein between, of liposomal trans-crocetin, followed by the administration of one or more fixed maintenance doses of 75 mg to 350 mg (e.g., 100 mg to 350 mg, 100 mg to 200 mg, 150 mg to 200 mg, 120 mg to 160 mg, or 140 mg), or any range therein between, of liposomal trans-crocetin. In some embodiments, two fixed loading dose of liposomal trans-crocetin are administered to a subject within 24 hours of each other. In some embodiments, two or more fixed maintenance doses of liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In particular embodiments, a fixed maintenance dose of liposomal trans-crocetin is administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In one embodiment, a subject is administered a fixed dose of 100 mg to 350 mg (e.g., 250 mg or 300 mg), or any range therein between of liposomal trans-crocetin, followed by the administration of a fixed dose of 100 mg to 350 mg (e.g., 120 mg to 160 mg, 140 mg, 250 mg or 300 mg), or any range therein between, of liposomal trans-crocetin. In particular embodiments, the subject is administered two or more fixed dose of 100 mg to 350 mg, or any range therein between of liposomal trans-crocetin within 24 hours of each other. In some embodiments, a fixed dose of 100 mg to 350 mg, or any range therein between, of liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, a fixed dose of 100 mg to 350 mg, or any range therein between, of liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more. In particular embodiments, two fixed dose of 100 mg to 350 mg, or any range therein between of liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In particular embodiments, two fixed dose of 100 mg to 350 mg, or any range therein between of liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In one embodiment, a subject is administered a fixed dose (loading dose) of approximately 225 mg to 275 mg (e.g., 250 mg) of liposomal trans-crocetin is followed by the administration of a fixed dose of approximately 120 mg to 160 mg (e.g., 140 mg))(maintenance dose) of liposomal trans-crocetin. In particular embodiments, two fixed loading dose of liposomal trans-crocetin are administered to a subject within 24 hours of each other. In some embodiments, the fixed maintenance doses of liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, the fixed maintenance doses of liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In one embodiment, a subject is administered a fixed dose (loading dose) of approximately 250 mg of liposomal trans-crocetin is administered to a subject followed by the administration of a plurality of fixed doses of approximately 140 mg (maintenance dose) of liposomal trans-crocetin. In particular embodiments, two fixed loading doses of liposomal trans-crocetin are administered to the subject within 24 hours. In some embodiments, the fixed maintenance doses of liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments, the fixed maintenance doses of liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In another embodiment, a plurality of fixed doses of 75 mg to 300 mg, or any range therein between, of liposomal trans-crocetin are administered to a subject, once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In particular embodiments, two fixed doses of 75 mg to 300 mg, or any range therein between, liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In another embodiment, a plurality of fixed doses of 100 mg to 250 mg, or any range therein between, of liposomal trans-crocetin are administered to a subject, once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In particular embodiments, two fixed doses of 100 mg to 250 mg, or any range therein between, liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
In another embodiment, a plurality of fixed doses of 300 mg to 400 mg, or any range therein between, of liposomal trans-crocetin are administered to a subject, once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In particular embodiments, two fixed doses of 300 mg to 400 mg, or any range therein between, liposomal trans-crocetin are administered to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month, for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or 1, 2, 3, 4, 5, or 6 years, or more.
Suitably, maintenance doses of the compositions provided herein maintain a relatively constant therapeutic level of trans-crocetin in a subject throughout the maintenance phase. The period during which maintenance doses are provided will depend on the length of time in which it is desired to maintain therapeutic levels of the liposomal trans-crocetin. Suitably the maintenance doses are distributed at regular intervals over the duration of treatment. In a suitable embodiment the time intervals between the maintenance doses may be longer than time intervals between the loading doses.
In one embodiment, the concentration of a fixed maintenance dose of a liposomal trans-crocetin composition provided herein is 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 80 mg to 275 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg, or 225 mg to 275 mg (e.g., 250 mg), or any range therein between, and the time interval between the administration of maintenance doses to a subject is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In another embodiment, the concentration of two or more fixed maintenance dose of liposomal trans-crocetin is 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), or any range therein between, and the time interval between the administration of maintenance doses to a subject once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
The trans-crocetin pharmaceutical compositions provided herein such as liposomal trans-crocetin compositions, have uses that provide advances over prior treatments of disorders and conditions that include without limitation, infection 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 kidney failure and ARDS, sepsis, inflammation (e.g., chronic inflammatory diseases), ischemia, (including an ischemic condition such as ischemic stroke, coronary artery disease, peripheral vascular disease, cerebral vascular disease, ischemia associated renal pathologies, and ischemia associated with wounds); shock (e.g., hemorrhagic shock), stroke, cardiovascular disease, renal pathologies, wound healing, metabolic disease, hyperproliferative diseases such as cancer, and disorders of the immune system, cardiovascular system, digestive, nervous, respiratory, and endocrine system. In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. Use of a pharmaceutical composition provided herein (e.g., the pharmaceutical composition of any of a fixed dose of trans-crocetin (e.g., a trans-crocetin dose(s) and/or dosing regimen(s) administered according to the method of any one of [1]-[65]), in the manufacture of a medicament for the treatment of aging and/or a chronic disease in a subject is also provided herein. As are, pharmaceutical compositions of any of a fixed dose of trans-crocetin (e.g., a trans-crocetin dose(s) and/or dosing regimen(s) administered according to the method of any one of [1]-[65]) for use in a medical medicament.
In one embodiment, the disclosure provides trans-crocetin pharmaceutical compositions and dosing regimens for use in treating an ischemic or hypoxic condition in a subject that comprises administering to the subject an effective amount of a liposomal trans-crocetin composition and/or dosing regimen provided herein (e.g., dosing regimen for a fixed dose of liposomal trans-crocetin), thereby treating an ischemic or hypoxic condition in the subject.
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition, for use in treating an ischemic or hypoxic condition in a subject, wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more loading doses of liposomal trans-crocetin in a fixed dose of 100 mg to 550 mg (e.g., 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 550 mg, 150 mg to 200 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg), and wherein all loading doses are administered within 12 hours (+/−3 hours) or 24 hours (+/−6 hours), and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of liposomal trans-crocetin in an amount of 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 80 mg to 275 mg, 100 mg to 200 mg, 120 mg to 160 mg (e.g., 140 mg), 200 mg to 300 mg, or 225 mg to 275 mg (e.g., 250 mg), or any range therein between, and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition, for use in treating an ischemic or hypoxic condition in a subject, wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more fixed loading doses of liposomal trans-crocetin in an amount of 100 mg to 350 mg, or any range therein between, and wherein all loading doses are administered within 3 hours, and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of liposomal trans-crocetin in an amount of 100 mg to 350 mg, or any range therein between and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in treating COPD in a subject, wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more fixed loading doses of liposomal trans-crocetin in (a) fixed dose of 100 mg to 550 mg (e.g., 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 550 mg, 150 mg to 200 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg), and wherein each loading doses is administered via infusion over 1-3 hours, and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of the liposomal trans-crocetin in a fixed dose of 100 mg to 550 mg (e.g., 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 550 mg, 150 mg to 200 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg) and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in treating COPD in a subject, wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more loading doses of liposomal trans-crocetin in a fixed dose of 100 mg to 550 mg (e.g., 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 550 mg, 150 mg to 200 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg), and wherein all loading doses are administered within 12 hours (+/−3 hours) or 24 hours (+/−6 hours), and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of the liposomal trans-crocetin in a fixed dose of 100 mg to 400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg) and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in treating COPD in a subject, wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more fixed loading doses of liposomal trans-crocetin in an amount of 100 mg to 350 mg, or any range therein between, and wherein all loading doses are administered within 3 hours, and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of liposomal trans-crocetin in an amount of 100 mg to 350 mg, or any range therein between and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in treating sepsis in a subject, wherein the fixed dose liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more loading doses of trans-crocetin in a fixed dose of 100 mg to 550 mg (e.g., 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 550 mg, 150 mg to 200 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg), or any range therein between, and wherein each loading dose is administered by infusion over 1 to 3 hours and liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of liposomal trans-crocetin at a fixed dose of 100 mg to 460 mg (e.g., 100 mg to 400 mg 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 200 mg 120 mg to 160 mg, 300 mg, 250 mg, or 140 mg), and wherein the time interval between 1, 2, 3, 4, 5, or more, or all, maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in treating sepsis in a subject, wherein the fixed dose liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or doses of liposomal trans-crocetin in a fixed dose of 100 mg to 460 mg (e.t250 mg or 300 mg), or any range therein between, is administered by infusion over 1 to 3 hours and wherein the subject is further administered a plurality of doses of liposomal trans-crocetin at a fixed dose of 100 mg to 350 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 200 mg 120 mg to 160 mg), or any range therein between, and wherein the time interval between 1, 2, 3, 4, 5, or more, or all, maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in treating a chronic infection in a subject, wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more loading doses of liposomal trans-crocetin at a fixed dose of 100 mg to 550 mg (e.g., 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 550 mg, 150 mg to 200 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg), and wherein each loading doses is administered via infusion over 1-3 hours, and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of the liposomal trans-crocetin in (a) a fixed dose of 100 mg to 400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg) and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments the infection is a bacterial infection (e.g., a chronic infection caused by Enterobacteriaceae species (spp.), Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, or Pseudomonas aeruginosa), a viral infection (e.g., a chronic infection caused by an influenza virus, or a coronavirus such as COVID-19), or a fungal infection. In particular embodiments the infection is caused by a virus (e.g., COVID-19).
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in treating a chronic infection in a subject, wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more loading doses of liposomal trans-crocetin at a fixed dose of 100 mg to 550 mg (e.g., 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 550 mg, 150 mg to 200 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg), and wherein all loading doses are administered within 12 hours (+/−3 hours) or 24 hours (+/−6 hours), and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of the liposomal trans-crocetin in a fixed dose of 100 mg-400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg) and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments the infection is a bacterial infection (e.g., a chronic infection caused by Enterobacteriaceae species (spp.), Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, or Pseudomonas aeruginosa), a viral infection (e.g., a chronic infection caused by an influenza virus, or a coronavirus such as COVID-19), or a fungal infection. In particular embodiments the infection is caused by a virus (e.g., COVID-19).
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in treating a chronic infection in a subject, wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more fixed loading doses of liposomal trans-crocetin in an amount of 100 mg to 350 mg, or any range therein between, and wherein all loading doses are administered within 3 hours, and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of liposomal trans-crocetin in an amount of 100 mg to 350 mg, or any range therein between and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month. In some embodiments the infection is a bacterial infection (e.g., a chronic infection caused by Enterobacteriaceae species (spp.), Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, or Pseudomonas aeruginosa), a viral infection (e.g., a chronic infection caused by an influenza virus, or a coronavirus such as COVID-19), or a fungal infection. In particular embodiments the infection is caused by a virus (e.g., COVID-19).
In one embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in increasing the delivery of oxygen in a subject, wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more loading doses of liposomal trans-crocetin at a fixed dose of 100 mg to 550 mg (e.g., 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 550 mg, 150 mg to 200 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg), and wherein each loading doses is administered via infusion over 1-3 hours, and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of the liposomal trans-crocetin in (a) a fixed dose of 100 mg-400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg) and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in increasing the delivery of oxygen in a subject wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more loading doses of liposomal trans-crocetin at a fixed dose of 100 mg-400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg), and wherein all loading doses are administered within 12 hours (+/−3 hours) or 24 hours (+/−6 hours), and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of the liposomal trans-crocetin in a fixed dose of 100 mg-400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg) and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in increasing the delivery of oxygen in a subject, wherein the liposomal trans-crocetin composition is first administered to a subject in a loading phase, during which the subject is administered 1, 2, 3, or more fixed loading doses of liposomal trans-crocetin in an amount of 100 mg to 350 mg, or any range therein between, and wherein all loading doses are administered within 3 hours, and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of liposomal trans-crocetin in an amount of 100 mg to 350 mg, or any range therein between and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In additional embodiments, the disclosure provides a method for increasing the delivery of oxygen in a subject who has or is at risk for developing ischemia, that comprises administering to the subject an effective amount of a liposomal trans-crocetin composition provided herein, such as a fixed dose liposomal trans-crocetin composition, thereby increasing the delivery of oxygen to the tissues and/or organs in the subject. In some embodiments, the subject has or is at risk for developing ischemia. In some embodiments, an effective amount of the pharmaceutical composition is administered to the subject before, during or following surgery (e.g., transplantation; reattachment of severed extremities, body parts or soft tissues; graft surgery, and vascular surgery). In some embodiments, an effective amount of the pharmaceutical composition is administered to a subject who has or is at risk for developing a wound, a burn injury, an electrical injury, or exposure to ionizing radiation. In some embodiments, an effective amount of the pharmaceutical composition is administered to a subject who has or is at risk for developing peripheral vascular disease, coronary artery disease, stroke, thrombosis, a clot, chronic vascular obstruction or vasculopathy (e.g., secondary to diabetes, hypertension, or peripheral vascular disease), or cerebral ischemia, pulmonary hypertension (adult or neonate); sickle cell disease; neointimal hyperplasia or restenosis (following angioplasty or stenting). In some embodiments, an effective amount of the pharmaceutical composition is administered to a subject who has or is at risk for developing a myopathy, kidney disease; asthma or adult respiratory distress syndrome; Alzheimer's and other dementias secondary to compromised cranial blood flow. In some embodiments, the method comprises administering a fixed dose of trans-crocetin (e.g., a trans-crocetin dose(s) and/or dosing regimen(s) administered according to the method of any one of [1]-[65]) to the subject. Use of a pharmaceutical composition provided herein such as a fixed dose liposomal trans-crocetin composition, in the manufacture of a medicament for increasing the delivery of oxygen in a subject is also provided herein. As are, pharmaceutical compositions of a fixed dose of trans-crocetin (e.g., a trans-crocetin dose(s) and/or dosing regimen(s) administered according to the method of any one of [1]-[65])] for use in a medical medicament. In some embodiments, the administered pharmaceutical composition comprises a surface active copolymer. In further embodiments, the liposomal composition comprises a poloxamer such as P188, P124, P182, P188, or P234. In yet further embodiments, the liposomal composition comprises the poloxamer P188.
Methods are also disclosed herein for increasing the delivery of oxygen in a neonate subject or a subject who is elderly that comprises administering to the subject a pharmaceutical composition provided herein, such as a fixed dose liposomal trans-crocetin composition, thereby increasing the delivery of oxygen to the tissues and/or organs of the subject. In some embodiments, the subject is elderly (e.g., a human subject that is more than 65, more than 70, more than 75, or more than 80 years of age). In some embodiments, the subject has or is at risk for developing a respiratory condition or disease (e.g., COPD, respiratory distress syndrome or adult respiratory distress syndrome). In some embodiments, the subject has or is at risk for developing a degenerative disorder, such as dementia or Alzheimer's disease. In some embodiments, the method comprises administering a fixed dose of trans-crocetin (e.g., a trans-crocetin dose(s) and/or dosing regimen(s) administered according to the method of any one of [1]-[65]) to the subject. Use of a pharmaceutical composition provided herein (such as a fixed dose liposomal trans-crocetin composition) in the manufacture of a medicament for increasing the delivery of oxygen in an elderly subject is also provided herein. As are fixed dose liposomal trans-crocetin compositions, for use in a medical medicament.
In additional embodiments, the disclosure provides a method for increasing the delivery of oxygen in a subject who has or is at risk for developing ischemia/reperfusion injury, that comprises administering to the subject an effective amount of a pharmaceutical composition pharmaceutical composition provided herein, such as a fixed dose liposomal trans-crocetin composition, thereby increasing the delivery of oxygen to the tissues and/or organs in the subject. In some embodiments, an effective amount of the pharmaceutical composition is administered to the subject before, during or following surgery (e.g., transplantation; reattachment of severed extremities, body parts or soft tissues; graft surgery, and vascular surgery). In some embodiments, the ischemia/reperfusion injury is due to a condition selected from infarction, atherosclerosis, thrombosis, thromboembolism, lipid-embolism, bleeding, stent, surgery, angioplasty, end of bypass during surgery, organ transplantation, total ischemia, and combinations thereof. In some embodiments, the ischemia/reperfusion injury is produced in an organ or a tissue selected from the group: heart, liver, kidney, brain, intestine, pancreas, lung, skeletal muscle and combinations thereof. In some embodiments, the ischemia/reperfusion injury is selected from the group: organ dysfunction, infarct, inflammation, oxidative damage, mitochondrial membrane potential damage, apoptosis, reperfusion-related arrhythmia, cardiac stunning, cardiac lipotoxicity, ischemia-derived scar formation, and combinations thereof. In particular embodiments, the ischemia/reperfusion injury is due to myocardial infarction. In some embodiments, an effective amount of the pharmaceutical composition is administered to a subject who has or is at risk for developing peripheral vascular disease, coronary artery disease, stroke, thrombosis, a clot, chronic vascular obstruction or vasculopathy (e.g., secondary to diabetes, hypertension, or peripheral vascular disease), or cerebral ischemia, pulmonary hypertension (adult or neonate); sickle cell disease; neointimal hyperplasia or restenosis (following angioplasty or stenting). In some embodiments, an effective amount of the pharmaceutical composition is administered to a subject who has or is at risk for developing a myopathy, kidney disease; asthma or adult respiratory distress syndrome; Alzheimer's and other dementias secondary to compromised cranial blood flow. In some embodiments, the method comprises administering a fixed dose of trans-crocetin (e.g., a trans-crocetin dose(s) and/or dosing regimen(s) administered according to the method of any one of [1]-[65]) to the subject. Use of a pharmaceutical composition pharmaceutical composition provided herein, such as a fixed dose liposomal trans-crocetin composition, in the manufacture of a medicament for increasing the delivery of oxygen in a subject is also provided herein. As are, a fixed dose of trans-crocetin (e.g., a trans-crocetin dose(s) and/or dosing regimen(s) administered according to the method of any one of [1]-[65]) for use in a medical medicament.
In an additional embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in increasing the efficacy of a therapeutic agent in a subject, that comprises administering to a subject who is receiving, will receive, or has received treatment with therapeutic agent, liposomal trans-crocetin in a loading phase during which the subject is administered 1, 2, 3, or more fixed loading doses of liposomal trans-crocetin in an amount of 100 mg to 350 mg, or any range therein between, and wherein all loading doses are administered within 3 hours, and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of liposomal trans-crocetin in an amount of 100 mg to 350 mg, or any range therein between and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in increasing the efficacy of a therapeutic agent in a subject, that comprises administering to a subject who is receiving, will receive, or has received treatment with therapeutic agent, liposomal trans-crocetin in a loading phase during which the subject is administered 1, 2, 3, or more loading doses of liposomal trans-crocetin at a fixed dose of 100 mg to 550 mg (e.g., 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 550 mg, 150 mg to 200 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg), and wherein each loading doses is administered via infusion over 1-3 hours, and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of the liposomal trans-crocetin in (a) a fixed dose of 100 mg-400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg) and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In another embodiment, the disclosure provides a fixed dose liposomal trans-crocetin composition for use in increasing the efficacy of a therapeutic agent in a subject, which comprises administering to a subject who is receiving, will receive, or has received treatment with therapeutic agent, liposomal trans-crocetin in a loading phase, during which the subject is administered 1, 2, 3, or more loading doses of liposomal trans-crocetin at a fixed dose of 100 mg to 550 mg (e.g., 100 mg to 460 mg, 100 mg to 350 mg, 100 mg to 300 mg, 100 mg to 200 mg, 150 mg to 550 mg, 150 mg to 200 mg, 225 mg to 275 mg, 140 mg, 250 mg, 300 mg, 380 mg, or 580 mg), and wherein each loading doses is administered via infusion over 1-3 hours, and wherein liposomal trans-crocetin is then further administered to the subject in a maintenance phase, during which the subject is administered a plurality of fixed maintenance doses of the liposomal trans-crocetin in (a) a fixed dose of 100 mg-400 mg (e.g., 100 mg to 300 mg, 100 mg to 200 mg, 120 mg to 160 mg, 300 mg, or 140 mg) and wherein the time interval between 1, 2, 3, 4, 5, or more, or all maintenance doses is once a day, twice a week, once a week, four times a month, three times a month, two times a month, or once a month.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with endotoxemia in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with sepsis in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. In some embodiments, the subject has a low grade endotoxemic disease.
In some embodiments, the disclosure provides a method for treating a subject at risk of developing sepsis, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) 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 elderly or neonatal. In some embodiments, the subject has febrile neutropenia. In some embodiments, the subject has a chronic infection.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with burn injury in a subject that is a burn victim, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with infection in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. In some embodiments, the infection is a bacterial infection (e.g., a P. aeruginosa infection, an S. aureus infection (e.g., MRSA), Mycobacterium tuberculosis infection, an enterococcal 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 kidney failure and ARDS. In some embodiments, the infection is a viral infection (e.g., COVID-19, Ebola, Dengue and Marburg) or a condition associated therewith, such as ARDS, influenza, measles, and a viral hemorrhagic fever.
In some embodiments, the disclosure provides trans-crocetin compositions and dosing regimens for treating anemia in a subject which comprises administering to a subject who has experienced, is experiencing, will experience, or is at risk of experiencing anemia, a liposomal trans-crocetin composition in a dosing regimen according to the method of any one of [1]-[65]). In some embodiments, trans-crocetin is administered to a subject having acute blood loss anemia (e.g., anemia caused by rapid massive hemorrhage) or an associated condition. In some embodiments, trans-crocetin is administered to a subject having chronic blood loss anemia (e.g., anemia caused by prolonged moderate blood loss or blood deficiency) or an associated condition.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with shock in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. 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. In particular embodiments, the administration of the liposomal trans-crocetin is associated with a reduction in heart rate, blood acidosis, and/or organ damage in the subject. In particular embodiments, trans-crocetin is administered within 1 hour or within 4, 12, 18 or 24 hours, or 48 hours of the onset of shock or a condition associated with shock.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with nitric oxide deficiency in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin (e.g., a trans-crocetin dose(s) and/or dosing regimen(s) administered according to the method of any one of [1]-[65]) to the subject. In some embodiments, the disease or disorder is sickle cell disease, paroxysmal nocturnal hemoglobinuria (PNH), a hemolytic anemia, a thalassemia, another red blood cell disorder, or a condition associated therewith. In some embodiments, the disease or disorder is a purpura such as thrombotic thrombocytic purpura (TTP), hemolytic uremic syndrome (HUS), idiopathic thrombocytopenia (ITP), or and another platelet disorder, or a condition associated therewith. In some embodiment, the disease or disorder is a coagulation abnormality such as disseminated intravascular coagulopathy (DIC), purpura fulminans, heparin induced thrombocytopenia (HIT), hyperleukocytosis, hyper viscosity syndrome, or a condition associated therewith.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with inflammation in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) 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.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with a cardiovascular disease or condition in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. In some embodiments, cardiovascular disease or condition is coronary artery disease. In some embodiments the cardiovascular disease or condition is myocardial infarction, sudden cardiac death, cardiorespiratory arrest, hypertension, pulmonary arterial hypertension, atherosclerosis, occlusive arterial disease, Raynaud's disease, peripheral vascular disease, other vasculopathies such as Buerger's disease, Takayasu's arthritis, and post-cardiac arrest syndrome (PCAS), chronic venous insufficiency, heart disease, congestive heart failure, or a chronic skin ulcer. Methods and biomarkers for evaluating cardiovascular health (e.g., levels of conventional troponins (cTnI and cTnT), Ischemia-Modified Albumin (IMA), B-type Natriuretic Peptide and N-terminal proBNP, whole blood choline, and unesterified free fatty acid (FFAu)) and cardiovascular injury and disease, and the efficacy of treatment regimens are known in the art
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with a liver disease, injury or condition in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. In some embodiments, the liver disease or condition is hepatic ischemia/reperfusion injury. In some embodiments, the liver disease or condition is a hepatic resection or liver transplantation. In some embodiments, the liver disease or condition is cirrhosis. In some embodiments, the liver disease or condition is nonalcoholic 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. Methods and biomarkers for evaluating liver health (e.g., levels of liver enzymes ALT, AST, ALP, and LDH), as well as liver injury and disease and the efficacy of treatment regimens are known in the art.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with a lung disease or condition in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) 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 (COPD). In some embodiments, the lung disease or condition is pulmonary fibrosis. In some embodiments, the lung disease or condition is emphysema. In some embodiments, the lung disease or condition is asthma. 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 (e.g., acute lung injury (ALI)). In some embodiments, the lung disease or condition is lung cancer. In some embodiments, the condition is cystic fibrosis.
In some embodiments, an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) is administered to a subject (e.g., a human) experiencing acute lung distress (e.g., presenting symptoms such as having difficulty breathing, tachypnea, mental confusion due to low oxygen levels) and/or having a PaO2/FiO2 ratio of less than 300 mm Hg or less than 250 mm Hg. In additional embodiments, the pharmaceutical composition is administered to a subject having a PaO2/FiO2 ratio of <300 mm Hg to ≥200 mm Hg. In additional embodiments, the pharmaceutical composition is administered to a subject having a PaO2/FiO2 ratio of <300 mm Hg to ≥250 mm Hg. In further embodiments the pharmaceutical composition comprises liposomal trans-crocetin.
In some embodiments, an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to a subject (e.g., a human) experiencing Acute Respiratory ARDS and/or having a PaO2/FiO2 ratio of less than 200 mm Hg. In further embodiments the pharmaceutical composition comprises liposomal trans-crocetin.
In some embodiments, an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to a subject (e.g., a human) in order to increase the patient's PaO2/FiO2 ratio. In some embodiments, administration of the pharmaceutical composition increases the patient's PaO2/FiO2 ratio by at 5%-75%, or any range therein between. In further embodiments, the administration of the pharmaceutical composition increases the patient's PaO2/FiO2 ratio by at least 5%, 10%, 15%, 20%, 25%, 30% 40% or 50%. In further embodiments the pharmaceutical composition comprises liposomal trans-crocetin. In further embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 5%-75%, or any range therein between. In further embodiments, the administration of the liposomal trans-crocetin increases the patient's PaO2/FiO2 ratio by 10%-50%, or any range therein between. In further embodiments, the administration of the trans-crocetin increases the patient's PaO2/FiO2 ratio by at least 5%, 10%, 15%, 20%, 25%, 30% 40% or 50%.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with a kidney disease or condition in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) 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 kidney disease. Methods and biomarkers for evaluating renal health (e.g., levels of N-acetyl-μ-glucosaminidase (NAG), α1-microglobulin (α1M), Cystatin-C (Cys-C), Retinol binding protein (RBP), microalbumin, Kidney injury molecule-1 (KIM-1), Clusterin, Interleukin-18 (IL18), Cysteine-rich protein (Cyr61), osteopontin (OPN), Fatty acid-binding protein (FABP), Fetuin-A, and neutrophil gelatinase-associated lipocalin (NGAL), as well as renal injury and disease and the efficacy of treatment regimens are known in the art.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with a vascular disease in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) 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 disorder or condition associated with a vascular disease is heart disease. In further embodiments, the disorder or condition is congestive heart failure. In some embodiments, the disorder or condition associated with vascular disease is ischemic bowel disease.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with a heart attack or stroke in a subject needing such treatment or prevention and/or at risk of having a heart attack or stroke, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. In some embodiments, the disorder or condition is ischemic stroke. In some embodiments, the disorder or condition is hemorrhagic stroke. Methods and biomarkers for evaluating heart attack and stroke (e.g., levels of blood B-type natriuretic peptide (BNP), C-reactive protein (CRP), GlycA, CK-MB, Cardiac troponin, myoglobin, low-density lipoprotein-cholesterol and hemoglobin A1c (HgA1c), lipoprotein-associated phospholipase A2, glial fibrillary acidic protein, S100b, neuron-specific enolase, myelin basic protein, interleukin-6, matrix metalloproteinase (MMP)-9, D-dimer, and fibrinogen), and the efficacy of treatment regimens are known in the art.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with nervous system in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) 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 disorder or condition associated with nervous system is neural injury.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with inflammatory bowel disease in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. In some embodiments, the disorder or condition is Crohn's disease. In some embodiments, the disorder or condition is ulcerative colitis.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with type 2 diabetes or predisposition for diabetes in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. In some embodiments, the disorder or condition is metabolic disease. In some embodiments, the disorder or condition is insulin resistance. In some embodiments, the disorder or condition is a diabetic vascular disease (e.g., a microvascular disease such as retinopathy and nephropathy). In some embodiments, the disorder or condition is diabetic neuropathy. In some embodiments, the disorder or condition is ulcers, diabetic necrosis, or gangrene.
In some embodiments, the disclosure provides a method for treating a myopathy, chronic microvascular disease, or microangiopathy, or a disorder associated with microvascular dysfunction such as age-related macular degeneration (AMD) in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject.
In some embodiments, the disclosure provides a method for treating aging and/or a chronic disease associated with sclerosis in a subject needing such treatment or prevention, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. In some embodiments, the disorder or condition associated with sclerosis is systemic sclerosis.
In some embodiments, the disclosure provides a method for treating endotoxemia in a subject needing such treatment, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject. In some embodiments, the endotoxemia is associated with a condition such as periodontal disease (e.g., periodontitis or inflammation of the gums), chronic alcoholism, chronic smoking, transplantation, or neonatal necrotizing enterocolitis, or neonatal ear infection.
In some embodiments, the disclosure provides a method of reducing systemic levels of LPS, endotoxin and/or another trigger of systemic inflammation in a subject in need thereof, the method comprising administering an effective amount of a pharmaceutical composition provided herein (e.g., a fixed dose of trans-crocetin in a trans-crocetin dose and/or dosing regimen administered according to the method of any one of [1]-[65]) to the subject.
The trans-crocetin compositions and dosing regimens provided herein can be administered alone or in combination therapy with one or more additional therapeutic agents. In some embodiments, the fixed dose liposomal trans-crocetin composition is administered in combination therapy with another therapeutic agent. Combinations may be administered either concomitantly, e.g., combined in the same liposomal composition, delivery vehicle (e.g., liposome), as an admixture, separately but simultaneously or concurrently; or sequentially. This includes presentations in which the combined therapeutic agents are administered together as a therapeutic mixture, and also procedures in which the combined agents are administered separately but simultaneously, e.g., as through separate intravenous lines into the same subject. Administration “in combination” further includes the separate administration of the liposomal trans-crocetin composition before or after the additional therapeutic agent(s). Methods of treatment using the combination therapy are also provided. In some embodiments, one or more doses of trans-crocetin is administered to the subject before the subject is administered the additional therapeutic agent (e.g., 5 minutes to 72 hours, 15 minutes to 48 hours, or 30 minutes to 24 hours before administration of the additional therapeutic agent, or within 12 hours, 9 hours, 6 hours, 4 hours, 2 hours, or within 1 hour before the administration of the additional therapeutic agent). In further embodiments, the additional therapeutic agent is radiation, a chemotherapeutic agent, an immunotherapeutic agent, or oxygen therapy.
The trans-crocetin pharmaceutical compositions and dosing regimens provided herein have applications in cancer therapy both as mono and combination therapy. In some embodiments, a liposomal trans-crocetin composition and/or dosing regimen provided herein is administered in combination therapy with one or more chemotherapeutic agents (e.g., to enhance the effect of chemotherapy on cancer cells and mitigate the effects of chemotherapy-induced myelosuppression and anemia). The combination therapy may include, for example, coadministration or concurrent administration, using separate formulations or a single pharmaceutical formulation, and consecutive administration in either order, wherein preferably there is a time period while both (or all) active agents simultaneously exert their biological activities. Thus, the chemotherapeutic agent may be administered prior to, or following, administration of the trans-crocetin. In this embodiment, the timing between at least one administration of the chemotherapeutic agent and at least one administration of the liposomal trans-crocetin is preferably approximately 1 week, 3 days, 24 hours, 12 hours, 6 hours or less. In some embodiments, the fixed dose liposomal trans-crocetin composition is administered to the subject before the chemotherapeutic agent (e.g., 5 minutes to 72 hours, 15 minutes to 48 hours, or 30 minutes to 24 hours before administration of the chemotherapeutic agent, or within 12 hours, 9 hours, 6 hours, 4 hours, 2 hours, or within 1 hour before the administration of the chemotherapeutic agent.
Alternatively, the chemotherapeutic agent and the trans-crocetin are administered concurrently to the patient, in a single formulation or separate formulations. Treatment with the combination of the chemotherapeutic agent and the trans-crocetin (e.g., liposomal trans-crocetin) may result in a synergistic, or greater than additive, therapeutic benefit to the subject.
In some embodiments, a liposomal trans-crocetin composition and/or dosing regimen 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 myelosuppression and anemia).
In some embodiments, a liposomal trans-crocetin composition and/or dosing regimen provided herein is administered in combination therapy with an immunotherapy. In some embodiments, one or more provided trans-crocetin compositions are administered to a subject before the administration of an immunotherapeutic agent (e.g., 5 minutes to 72 hours, 15 minutes to 48 hours, or 30 minutes to 24 hours before, or within 12 hours, 9 hours, 6 hours, 4 hours, 2 hours, or 1 hour before the administration of the immunotherapeutic agent.
In some embodiments, one or more provided trans-crocetin compositions are administered to a subject before the administration of radiation (e.g., 5 minutes to 72 hours, 15 minutes to 48 hours, or 30 minutes to 24 hours before, or within 12 hours, 9 hours, 6 hours, 4 hours, 2 hours, or 1 hour before the administration of the radiation.
In another embodiment, the disclosure provides a kit for administering a provided trans-crocetin composition (e.g., a fixed dose liposomal trans-crocetin composition) to a subject for treating a disorder, or condition. In some embodiments, the disclosure provides a kit for delivering a therapeutic agent to a subject, the kit comprising: (a) a first composition comprising a provided trans-crocetin composition (e.g., a fixed dose liposomal trans-crocetin composition); and a (b) second composition containing for example, reagents, buffers, excipients, or another therapeutic agent that is stored separately prior to administration to the subject. Such kits typically include two or more components necessary for treating a disease state, such as hypoxia or inflammation related condition. In some embodiments, the kits include for example, a provided fixed dose trans-crocetin composition, reagents, buffers, containers and/or equipment. The trans-crocetin compositions and formulations can be in lyophilized form and then reconstituted prior to administration. In some embodiments, the kits include a packaging assembly that include one or more components used for treating the disease state of a patient. For example, a packaging assembly may include separate containers that house therapeutic trans-crocetin compositions and other excipients or therapeutic agents that can be mixed with the compositions prior to administration to a patient. In some embodiments, a physician may select and match certain components and/or packaging assemblies depending on the treatment or diagnosis needed for a particular patient.
Two different variants of trans-crocetin were used to produce trans-crocetin liposomes, namely: trans-crocetin free acid (TC) and its sodium salt, sodium trans-crocetin (STC). Trans-crocetin was encapsulated in liposomes by the following procedures.
First, the lipid components of the liposome lipid membrane were weighed out and combined as a concentrated solution in ethanol at a temperature of around 65° C. In one preparation, the lipids used were hydrogenated soy phosphatidylcholine, cholesterol, and DSPE-PEG-2000 (1,2-distearoyl-sn-glycero-3-phosphoethan-olamine-N-[methoxy (polyethylene glycol)-2000]). The molar ratio of HSPC: cholesterol: PEG-DSPE was approximately 3:2:0.15. In another preparation, the lipids used were HSPC, cholesterol, PEG-DSPE-2000, and 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine (PGPC). The molar ratio of HSPC: cholesterol: PEG-DSPE:PGPC was approximately 2.7:2:0.15:0.3. Next, calcium acetate was dissolved in an aqueous buffer at a concentration of 125 mM, or 250 mM, with a pH of 7.0. The calcium acetate solution was heated up to 65° C.
The ethanolic lipid solution was added into the calcium acetate solution using a pipet. During this step the solution was well stirred using a magnetic stirrer. The mixing was performed at an elevated temperature (63° C.-72° C.) to ensure that the lipids were in a liquid crystalline state (as opposed to the gel state that they would attain at temperatures below the lipid transition temperature (Tm=51° C.-54° C.)). As a result, the lipids were hydrated and formed multiple bilayer (multilamellar) vesicles (MLVs) containing calcium acetate in the interior space.
The MLVs were fragmented into unilamellar (single bilayer) vesicles of the desired size by high-pressure extrusion using two passes through stacked (track-etched polycarbonate) membranes. The stacked membranes had two layers with a pore size of 200 nm and six layers with a pore size of 100 nm. During extrusion, the temperature was maintained above the Tm to ensure plasticity of the lipid membranes. As a result of the extrusion, large and heterogeneous in size and lamellarity MLVs were turned into small, homogenous (100-120 nm) unilamellar vesicles (ULVs) that sequestered calcium acetate in their interior space. A Malvern Zetasizer Nano ZS instrument (Southborough, MA) with back scattering detector (90°) was used for measuring the hydrodynamic size (diameter) of the vesicles at 25° C. in a plastic micro cuvette. The samples were diluted 50-fold in formulation matrix before analysis.
After ULVs containing calcium acetate had been produced, the extra-liposomal calcium acetate was removed using SEC (size exclusion chromatography, with PD10 columns) or TFF (tangential flow diafiltration). Tonicity reagent was added to the liposomes to balance the osmolality (final concentration: 5% dextrose for 125 mM calcium acetate liposomes and 10% dextrose in for 250 mM calcium acetate liposomes). Once the calcium acetate gradient was generated, the trans-crocetin loading procedure is preferably performed within 24 hours. The lipid content of the prepared liposome solution was determined by phosphate assay.
1 mg/mL trans-crocetin solution was prepared in 10% dextrose (for 250 mM calcium acetate liposomes) and pH was adjusted to 8. The trans-crocetin solution was mixed with 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 thoroughly stirred and heated to 65° C. for 30 minutes, followed by quick cool down to room temperature using an ice water bath. This step can be replaced by stirring the mixture at room temperature overnight.
The movement of trans-crocetin molecule (charge-free, neutral form) across the liposome lipid bilayer was driven by the gradient generated with calcium acetate (in other words, acetic acid diffused out, trans-crocetin diffused in). Trans-crocetin was then trapped inside of the liposomes by ionizing and then forming a precipitate with calcium (as a calcium salt form (calcium trans-crocetin, CTC)).
The extra-liposomal trans-crocetin was removed using SEC (PD10 columns) or TFF. In this example, the buffer used in SEC was HBS (HEPES buffered saline, pH 6.5). Upon completion of purification, filter sterilization was performed using a 0.22 micron filter. A Malvern Zetasizer Nano ZS instrument (Southborough, MA) with back scattering detector (90°) was used for measuring the hydrodynamic size (diameter) of the vesicles at 25° C. in a plastic micro cuvette. The samples were diluted before analysis.
Calcium acetate loaded liposomes 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 around 65° C. In one example, the lipids used were hydrogenated soy 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 was approximately 3:2:0.15. In another example, the lipids used were HSPC, cholesterol, PEG-DSPE-2000, and 1-palmitoyl-2-glutaryl-sn-glycero-3-phosphocholine (PGPC). The molar ratio of HSPC: cholesterol:PEG-DSPE:PGPC was approximately 2.7:2:0.15:0.3.
Next, calcium acetate was dissolved in an aqueous buffer at a concentration of 125 or 250 mM, with a pH of 7.0. The calcium acetate solution was heated to 65° C. The ethanolic lipid solution and the calcium acetate solution were separately transferred to syringes. Two solutions were injected into microfluidic channel and mixed while flowing through it with Precision NanoSystems' NanoAssemblr® device. The mixing was performed at an elevated temperature (63° C.-72° C.) to ensure that the lipids were in the liquid crystalline state (as opposed to the gel state that they would attain at temperatures below the lipid transition temperature (Tm=51° C.-54° C.)). The size of liposome can be controlled by ratio between lipid solution and aqueous solution, as well as the mixing flow rate.
A phase 2 clinical study was initiated to assess the safety and efficacy of liposomal trans-crocetin in patients with acute respiratory distress syndrome due to COVID-19 disease. The liposomal trans-crocetin compositions administered in this study were prepared essentially as described in international publication number WO2019213538, the contents of which are herein incorporated by reference in its entirety. The clinical study was an open label phase II study of treatment with liposomal trans-crocetin in patients who experience severe acute respiratory distress syndrome (ARDS) and were receiving artificial respiratory support due to COVID-19. The purpose of this study was to evaluate the improvement in PaO2/FiO2 by more than 25% in two cohorts of patients treated liposomal trans-crocetin. In the first cohort, patients received trans-crocetin at a dose of 0.25 mg/kg administered as intravenous (IV) bolus every 3 hours. The second cohort was treated by a liposomal formulation containing trans-crocetin. The liposomal formulation allowed for a gradual release of the free drug, to study the feasibility of once a day or less frequent dosing. Pharmacokinetic assessment was carried out to identify an optimal dose and schedule. Primary outcome measures included proportion of patients showing an increase of at least 25% of PaO2/FiO2 ratio (Time Frame: 24 hours). Secondary outcome measures included (1) proportion of patients with a PaO2/FiO2 ratio above 200 mm Hg (Time Frame: 24, 48 and 72 hours), and (2) all cause mortality (Time Frame: 28 days).
Data obtained from the first patients is disclosed herein. An additional 8-10 patients will be enrolled at the Cohort 4 dose to further demonstrate safety and efficacy.
The drug substance and active moiety tested was trans-crocetin and the drug product was liposomal trans-crocetin injection. Trans-crocetin, a carotenoid, has been shown to possess the capabilities to increase the diffusion of oxygen in plasma or water across a diffusion gradient. Gainer, Expert Opin. Investig. Drugs, 17(6):917-924 (2008); Gainer et al., Circ. Shock, 41(1):1-7 (1993); Roy et al., Shock, 10(3):213-217 (1998); Dhar et al., Mol. Cell. Biochem. 278(1-2):139-146 (2005).
Crocetins, as a class of natural products, have been recognized as having multiple clinically beneficial properties, including but not limited to, oxygen transport in hypoxic conditions and management of endotoxemia associated clinical conditions. Crocetins, in particular the sub-group of trans-crocetin salts and the one previously investigated, sodium trans-crocetin, which is also referred to as trans sodium crocetinate (TSC), have a unique property whereby they alter the structure of water in blood plasma by causing additional hydrogen bonds to form among the water molecules. As a consequence of this ‘order-making’ effect of trans-crocetin, also referred to as kosmotropic effect, trans-crocetin improves the diffusion of oxygen as well as other small molecules such as glucose, in plasma.
The clinical development of trans-crocetin has been problematic because its oxygen diffusion enhancing effect is transient, due to two key factors. Poor solubility: trans-crocetinate monovalent metal salts such as TSC, as free drugs were designed presumably to overcome the solubility issue associated with crocetin (free acid). Short half-life: TSC half-life is approximately 30 minutes, which in a clinical setting leads to transient effect on oxygen diffusion enhancement. The liposomal encapsulation of trans-crocetin generated a stable formulation with remarkably increased half-life (t½, 2 to 6 hours compared to half an hour for TSC in a mouse PK study).
Patients with severe COVID-19 frequently present with features of acute respiratory distress syndrome (ARDS). These patients usually require respiratory support with a ventilator. However, many patients present early on with severe hypoxia without extensive lung damage. Patients with this hypoxia, also termed “silent hypoxia”, may not necessarily show classic ARDS. Such patients compensate for hypoxia by breathing faster until they experience respiratory fatigue and collapse suddenly. In such patients, mechanical ventilation may not be as helpful, as has been evidenced by a relatively high mortality rate reported in patients receiving mechanical ventilation in the setting of COVID-19.
Without being bound by a specific theory, liposomal trans-crocetin could improve hypoxia in the body primarily in the following two ways: (1) Increase the rate of oxygen diffusion at the alveolar-capillary interface by increasing the diffusivity of oxygen across a gradient in conditions such as ARDS, and (2) Increase the rate of oxygen diffusion at the point where oxygen leaves the red blood cells and travels through plasma and interstitium to reach the tissues and individual cells. When this biological process of oxygen diffusion at this level is compromised, silent hypoxia could occur.
The primary purpose of this study was to determine a safe dosing regimen of liposomal trans-crocetin and to assess preliminary efficacy as measured by improvement in PaO2/FiO2 by more than 25% in patients treated with liposome encapsulated trans-crocetin, as well as safety and the pharmacokinetic profile of liposome encapsulated trans-crocetin.
Three dosing schedules have been studied so far in four cohorts of patients, as described in the table below. All doses were administered over 90 minutes, as an infusion intravenously.
While trans sodium crocetinate (TSC) was investigated in clinical trials with no safety concerns, the present study was the first clinical study to investigate a liposomal form of trans-crocetin.
In the first cohort treated with liposome encapsulated trans-crocetin, 6 patients were enrolled at the selected dosage of 2.5 mg/kg/day to determine if the dosing regimen was safe and whether it provided the required exposure for efficacy. Based on preclinical assessments, the target therapeutic range of free trans-crocetin should be between 0.4 and 49 μg/mL. Since 2 mg/kg/day was the dose safely administered in human for the primary ingredient, a 2.5 mg/kg/day dosage for liposomal trans-crocetin was selected on the fact that the encapsulated form is expected to release 90% of the overall free drug amount over time.
In the first cohort, six patients were treated with liposomal trans-crocetin at 2.5 mg/kg/day by an infusion of 90 minutes. Among the 30 cycles of administration no adverse event was observed. The PK of the total drug pointed out the need of 3 cycles to reach the point of balance suggesting the requirement of a loading dosage (
A second cohort of 6 patients was subsequently treated with a loading dose at Day 1 of 5 mg/kg followed by the 2.5 mg/kg/day (Table 1). The loading dosage allowed a target therapeutic range to be achieved from Day 1 and maintain the PK parameters in accordance with the previous findings from cohort 1 (Table 2). Again, no safety concern was observed. A stronger signal of activity emerged with the 5 mg/kg loading dose, as shown in
A subset of 4 patients with renal dysfunction who required dialysis were included in a third cohort. A different pharmacokinetic profile was observed in this subset of patients. PK for the total drug demonstrated a decrease in the volume of distribution in patients in Cohort 3. This was most likely attributable to a re-balance of drug between the patient plasma and the dialysate (Table 4).
The PK of the free drug from Day 1 in patients from Cohort 2 indicated that free drug concentrations were very low at H16 and H24 (ranged between 5.2-0.3 μg/mL) (
Two patients in cohort 4 was treated with the revised dosing schedule of 7.5 mg/kg loading dose followed by 5 mg/kg every 12 hours thereafter. The 2 patients from the cohort 4 experienced adverse events: grade II increase of total bilirubin without any hepatic biological disturbance. This suggests a possible interference between the color of trans-crocetin and the colorimetric method used to measure bilirubin could be the explanation and a measurement artefact. With respect to efficacy, both patients showed improvement of PaO2/FiO2 ratio and a decrease in the level and aggressiveness of artificial ventilation (as reflected by a decrease in positive expiratory pressure (PEP; cmH2O), and FIO2) in Cohort 2 over time (
After additional investigation based on various methods of assessment of bilirubin quantification, we concluded that the observed elevation of bilirubin was artefactual due to the interference between TC and the assay and protocols used for the quantification of bilirubin levels. No other AEs related to the administration of L4L-121 were reported (data not shown).
In cohort 4, the PK profile of the total drug was in accordance with the previous findings. The free drug concentration between doses appeared to remain within the boundaries of biological activity at H8 and H12 (ranged from 8.6 to 4.6 μg/mL>0.49 μg/mL). The Cmax following the loading dose were beyond the upper margin of activity (59 and 88 μg/mL>49 μg/mL) while the Cmax following the maintenance dosage were within the margins (34.7-44 μg/mL<49 μg/mL). Contamination by the liposome form may over-estimate the value of free drug concentration and may need to be taken into consideration.
In summary, the dosing of humans at the following doses is reported:
No adverse events or safety concerns were reported in Cohorts 1, 2 and 3. A Grade 2 elevation of bilirubin levels, without other corresponding hepatic abnormalities, was seen in both patients in Cohort 4. This raises the possibility that this might be artefactual and due to a possible interference between the color of trans-crocetin and the colorimetric method used to measure bilirubin.
With a day dosing, exposures observed revealed that the free trans-crocetin levels were undetectable in many patients at 8 hours for doses of 2.5 mg/kg, and at 12 hours for the loading dose of 5 mg/kg dose followed by 2.5 mg/kg a day. This suggested that, with a once a day dosing strategy used in Cohort 1, 2 and 3, many patients were being exposed to sub-therapeutic drug levels for at least half, if not most, of the treatment period.
PK exposure seen in patients with renal impairment suggested that such patients may require dose modifications specific to their clinical condition.
With a 7.5 mg/kg loading dose followed by 5 mg/kg every 12 hours thereafter studied in Cohort 4, preliminary data suggested that patients were being exposed to drug levels that decreased over time but remained within the therapeutic range during the course of treatment.
Despite the low exposures in Cohort 1, 2 and 3 patients, encouraging activity was noted with responses at 24 hours and beyond documented in all cohorts. Of note, in Cohort 2, all patients had a >25% improvement in PaO2/FiO2 ratio by 96 hours. These responses were staggered over time, perhaps likely due to the suboptimal PK exposure described above. In contrast, in Cohort 4, with a higher and sustained therapeutic drug exposure, >25% increase in PaO2/FiO2 ratio was observed at 24 hours in both patients enrolled in this cohort.
Efficacy in COVID-19 patients
The primary objective of this study was to demonstrate that treatment with L4L-121 leads to an improvement of 25% or more in PaO2/FiO2 in patients with ARDS under artificial respiratory support. Overall, 39% of the patients had an increase of >25% during the first 24 h of treatment with L4L-121. All patients in cohorts 2 and 3 and 50% of patients in cohorts 1 and 4 had a >25% improvement in their PaO2/FiO2 ratio anytime during the treatment period. Collectively, 78% of the patients had a >25% improvement in their PaO2/FiO2 ratio at any time during the treatment period (
For secondary indicators of efficacy, an overall 28-day survival rate of 83% was observed (
Although 1 patient had died by day 8, the number of patients requiring the most aggressive form of ventilatory support decreased from 13 (72%) patients at baseline to 8 (44%) patients by day 8, as did the requirement for paralytics and sedatives, in general. A strategy used to improve the ventilation of patients with ARDS is to place them in a prone position to increase oxygen diffusion. A total of 4 patients were prone at the start of treatment; however, by the end of the treatment at day 5, all patients were switched back to the supine position, improving their overall comfort (
Of interest, addressing parameters of oxygenation, the PaO2/FiO2 ratio tended to increase over time after treatment with L4L-121, with a more evident increase after day 2 post-treatment (
Overall, following treatment with L4L-121, a trend was observed where the mean total sequential organ failure assessment (SOFA) scores decreased by more than 2 points from baseline to day 8 (
In conditions such as COVID-19 and sepsis, ARDS leading to systemic oxygen deprivation or hypoxia has been identified as the primary cause of death, especially in severe cases. Hypoxia has been linked to a sequence of self-enhancing poor prognostic factors for clinical outcome in COVID-19 patients and other ARDS-related conditions, such as sepsis.
Treatment options for patients with severe COVID-19 are currently limited to mechanical ventilation and supportive care, with a poor survival outcome.
In vitro results and preclinical therapeutic assessment in a mouse model of sepsis demonstrated the reoxygenation activity of TC, indicating the enhancement of oxygen diffusion and the exposure optimization provided by the L4L-121 formulation. These preclinical data enabled us to perform a fast-track approved clinical study to evaluate this novel liposomal platform.
The patients enrolled in the study were on average hospitalized in the ICU for ˜12 days prior to treatment with underlying comorbidities that have been associated with worsened outcome by SARS-CoV-2 infection. Moreover, 100% of the patients included in the study demonstrated increased clotting activity, further emphasizing that the patient population treated was sick and would be expected to have challenges with organ perfusion.
Thirty-nine percent of the patients met the primary endpoint of a >25% improvement in the PaO2/FiO2 ratio at 24 h. In addition, up to 78% of the patients attained a >25% PaO2/FiO2 ratio during the treatment period. Beyond this ratio, additional markers of improvement in respiratory function during treatment included a general decrease in the FiO2 requirement and a slight decrease in PaCO2, a decrease in aggressive ventilation, a decrease in the use of sedatives and paralytic agents, and a decrease in the number of patients requiring placement in a prone position. Furthermore, the observed improvement in total SOFA score and subscores indicated that treatment with L4L-121 may improve microcirculation and thus perfusion of other organs, thereby enhancing multiorgan function in addition to the benefits observed in the respiratory system. Together, these observations suggest the potential clinical benefit of L4L-121 in treating patients with COVID-19 and, more broadly, in other high-unmet medical need settings with a similar underlying pathophysiology, such as sepsis, where multiorgan dysfunction is an important contributor to morbidity and mortality.
In this study, a 28-day survival rate of 83% was observed, which is particularly promising. Of note, in a similar patient population, survival rates of 57% and 69% were observed following treatment with standard care without and with dexamethasone, respectively (RECOVERY trial) 15. To date, no related AEs have been identified with the clinical use of L4L-121 at doses up to 12.5 mg/kg administered in a 24 h period. In conclusion, the overall risk/benefit profile of L4L-121 for the treatment of patients with ARDS due to COVID-19 requiring mechanical ventilation appears to be favorable in this high-risk patient population, which currently has no approved medications. The findings of this ongoing study, though promising, are preliminary and coupled with a small sample size. Nevertheless, given the urgent need in the context of the pandemic situation, a temporary authorization for routine use has been established in France for this compound since Nov. 17, 2020.
In vitro, we first sought to evaluate the reoxygenation properties of LEAF-4L121 in endothelial cells (HUVECs). The IC50 of LEAF-4L121 in HUVECs was determined after 72 h of treatment by viability assays (CellTiter-Glo, Promega). No toxicity was observed for concentrations up to 1 mg/mL, with an IC50 of 1136±21 μg/mL (
While the disclosed methods have been described in connection with what is presently considered to be the most practical and preferred embodiments, it is to be understood that the methods encompassed by the disclosure are not to be limited to the disclosed embodiments, but on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.
The disclosure of each of U.S. Appl. No. 62/666,699, filed May 3, 2018, U.S. Appl. No. 63/007,884, filed Apr. 9, 2020, U.S. Appl. No. 63/007,777, filed Apr. 9, 2020, U.S. Appl. No. 63/007,878, filed Apr. 9, 2020, U.S. Appl. No. 63/029,362, filed May 22, 2020, U.S. Appl. No. 63/029,362, filed May 24, 2020, herein incorporated by reference in its entirety.
The disclosure of each of Intl. Appl. Nos. PCT/2021/026703, PCT/2021/026704, PCT/2021/026683, PCT/2021/026532, PCT/2021/026654, and PCT/2021/026513, filed Apr. 9, 2021 and each of U.S. Appl. No. 63/071,312, filed Aug. 27, 2020, U.S. Appl. No. 63/064,300, filed Aug. 11, 2020, and U.S. Appl. No. 63/057,203, filed Jul. 27, 2020, is herein incorporated by reference in its entirety.
The disclosure of the international application entitled each of U.S. Appl. No. 63/057,208, filed Jul. 27, 2020, U.S. Appl. No. 63/057,203, filed Jul. 27, 2020, U.S. Appl. No. 63/063,809, filed Aug. 10, 2020, U.S. Appl. No. 63/064,281, filed Aug. 11, 2020, and U.S. Appl. No. 63/064,300, filed Aug. 11, 2020, is herein incorporated by reference in its entirety.
All publications, patents, patent applications, internet sites, and accession numbers/database sequences including both polynucleotide and polypeptide sequences cited herein are hereby incorporated by reference herein in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, internet site, or accession number/database sequence were specifically and individually indicated to be so incorporated by reference.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/US22/27595 | 5/4/2022 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 63186737 | May 2021 | US |
