Vitamin-based lipids and lipid nanoparticles comprising same

Abstract

Compounds, compositions, and methods for delivering therapeutic, diagnostic, or prophylactic agents (e.g., nucleic acids) are described.

Description

[0001] Cross - reference to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 420,245, filed on October 28, 2022, the content of which is hereby incorporated in its entirety. Background Art

[0003] The effective targeted delivery of active agents such as diagnostic agents, small molecule drugs, proteins, and nucleic acids represents an ongoing medical challenge. In particular, the delivery of nucleic acids to cells is difficult due to the relative instability and low cellular permeability of such species.

[0004] Lipid-containing nanoparticle compositions, liposomes, and lipid complexes have been shown to be effective as vehicles for the transport of active agents into cells and / or intracellular compartments. Despite the demonstration of a variety of such lipid-containing nanoparticle compositions, there remains a lack of improvement in terms of safety, efficacy, and specificity.

[0005] Accordingly, there is a need to develop methods and compositions that facilitate the delivery of therapeutic, diagnostic, and / or prophylactic agents such as nucleic acids to cells.

[0006] The compounds, compositions, and methods disclosed herein address these and other needs. Summary of the Invention

[0007] The present disclosure provides compounds and their uses. Compositions comprising the compounds and agents of the present invention are also provided. The present disclosure also provides methods of using the compositions to deliver active agents to a subject.

[0008] In one aspect, compounds of formula Da, Db, or Dc are described herein:

[0009]

[0010]

[0011] Wherein:

[0012] Z is a vitamin moiety;

[0013] X is O or S;

[0014] L 1 is -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-;

[0015] L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -NR a C(=O)- or -C(=O)R a N-;

[0016] L 3 is absent, and is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-;

[0017] R a is wherein n is an integer from 0 to 12;

[0018] x is 0, 1 or 2;

[0019] M is N, CH,

[0020] D is N or

[0021] G 1 , G 2 , G 3 and G 5 each independently selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0022] G 4 is absent or selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0023] R 2 is -CR b R c , C6-C 24 alkyl or C6-C 24 alkenyl;

[0024] R 3 is

[0025]

[0026] R 6 is H, OR b , CN, -C(=O)OR b , -NC(=O)R b, -C(=O)NR b ,

[0027] R b and R c are independently H, C1-C 12 alkyl or C1-C 12 alkenyl; and

[0028] p, q, and r are independently 0, 1, 2, 3, or 4;

[0029] or a pharmaceutically acceptable salt thereof.

[0030] In some embodiments, the vitamin moiety can be selected from:

[0031]

[0032] wherein

[0033] R 1 , R 4 and R 10 are each independently hydrogen or C1-C6 alkyl; and represents a carbon-carbon bond or a carbon-carbon double bond.

[0034] In some embodiments, compounds of formula Y, formula B, or formula C are described herein:

[0035]

[0036] wherein:

[0037] R 1 , R 4 and R 10 are each independently hydrogen or C1-C6 alkyl; represents a carbon-carbon bond or a carbon-carbon double bond;

[0038] A is

[0039]

[0040]

[0041] X is O or S;

[0042] L 1 is -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NRa C(=O)-;

[0043] L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -NR a C(=O)- or -C(=O)R a N-;

[0044] L 3 is absent, is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-;

[0045] R a is where n is an integer from 0 to 12;

[0046] x is 0, 1 or 2;

[0047] M is N, CH,

[0048] D is N or

[0049] G 1 , G 2 , G 3 and G 5 each independently selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0050] G 4 is absent or selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0051] R 2 is -CR b R c , C6-C 24 alkyl or C6-C 24 alkenyl;

[0052] R 3 is

[0053]

[0054] R 6 is H, OR b , CN, -C(=O)ORb 、 -NC(=O)R b 、 -C(=O)NR b 、

[0055] R b and R c are independently H, C1-C 12 alkyl or C1-C 12 alkenyl; and

[0056] p, q and r are independently 0, 1, 2, 3 or 4;

[0057] or a pharmaceutically acceptable salt thereof.

[0058] In some embodiments, compounds of Formulas I, II, III, IV, V, VI, VII, VIII or IX are described herein:

[0059]

[0060]

[0061]

[0062] Wherein:

[0063] R 1 、 R 4 and R 10 are each independently hydrogen or C1-C6 alkyl;

[0064] represents a carbon-carbon bond or a carbon-carbon double bond;

[0065] X is O or S;

[0066] L 1 is -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-;

[0067] L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -NR a C(=O)- or -C(=O)R a N-;

[0068] L 3 is absent, being -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-;

[0069] R a is wherein n is an integer from 0 to 12;

[0070] x is 0, 1 or 2;

[0071] M is N, CH,

[0072] D is N or

[0073] G 1 、G 2 、G 3 and G 5 each independently selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0074] G 4 is absent or selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0075] R 2 is -CR b R c 、C6-C 24 alkyl or C6-C 24 alkenyl;

[0076] R 3 is

[0077] R 6 is H, OR b 、CN, -C(=O)OR b 、-NC(=O)R b 、-C(=O)NR b 、

[0078]

[0079] R b and R c are independently H, C1-C 12 alkyl or C1-C12 an alkenyl group; and

[0080] p, q, and r are independently 0, 1, 2, 3, or 4;

[0081] or a pharmaceutically acceptable salt thereof.

[0082] Also described herein are lipid nanoparticles comprising greater than 0% to 50% of a compound described herein; 0% to 50% of a cationic lipid; 0% to 80% of an ionizable lipid; 0% to 5% of a polyethylene glycolated lipid; 0% to 40% of a helper lipid; 0% to 80% of a sterol; and an active agent encapsulated within the nanoparticles. In some embodiments, the lipid nanoparticles can comprise greater than 0% to 50% of a compound described herein; optionally greater than 0% to 50% of a cationic lipid; greater than 0% to 80% of an ionizable lipid; greater than 0% to 5% of a polyethylene glycolated lipid; greater than 0% to 40% of a helper lipid; greater than 0% to 80% of a sterol; or any combination thereof; and an active agent encapsulated within the nanoparticles.

[0083] Also described herein are compositions comprising a compound described herein or a lipid nanoparticle described herein and an active agent.

[0084] In some embodiments, provided herein are methods for delivering an agent (e.g., a polynucleotide). In some embodiments, provided herein are methods for diagnosing, treating, or preventing a disease. In some embodiments, provided herein are methods for inducing an immune response against an infectious agent. In some embodiments, provided herein are methods for treating cancer. In some embodiments, provided herein are methods for modulating the immune system to treat cancer and other immune disorders. BRIEF DESCRIPTION OF THE DRAWINGS

[0085] Figure 1 Shows the synthetic route of compound 4 (iVe).

[0086] Figures 2A - 2C Shows the 1 1H NMR spectrum (2A) of compound iVe, 13 13C NMR spectrum (2B), and mass spectrum (2C).

[0087] Figure 3 Shows the representative lipid nanoparticle (LNP) size distribution using iVe lipid formulated with eGFP-Luc mRNA.

[0088] Figures 4A - 4CShows the in vitro expression of GFP and luciferase after transfection of 293T cells with 1.0 μg of eGFP-Luc mRNA at 1.0 μg / mL mRNA for 24 hours. (4A) Representative fluorescence images of 293T cells after transfection. (4B) Transfection efficacy analysis of GFP expression using flow cytometry. (4C) Transfection effect of luciferase expression analyzed by bioluminescence.

[0089] Figures 5A - 5D Shows the in vivo transfection efficiency of mRNA expressing luciferase. The LNP was formulated with iVe lipid, and 1 μg of the formulated mRNA expressing luciferase was intramuscularly injected. After administration, (5A-5B) show representative images of luciferase expression determined by whole-body bioluminescence imaging of the draining lymph nodes (dLN) (5A) and injection site (5B) at 4, 24, and 48 hours using the IVIS Spectrum in vivo imaging system, and (5C-5D) show graphs of quantitative luciferase expression determined by whole-body bioluminescence imaging of the dLN (5C) and injection site (5D) at 4, 24, and 48 hours using the IVIS Spectrum in vivo imaging system. dLN represents the draining lymph node.

[0090] Figure 6 Shows the synthetic route of lipid iVd (T14).

[0091] Figures 7A - 7C For compound iVd 1 1H NMR spectrum (7A), mass spectrum (7B), and HPLC (7C).

[0092] Figures 8A - 8B Shows the in vitro expression of GFP after transfection of 293T cells with 1.0 μg of eGFP-Luc mRNA at 1.0 μg / mL mRNA for 24 hours. (8A) Transfection efficacy analysis of GFP expression using flow cytometry. (8D-8D) Representative fluorescence images of 293T cells after transfection (control (8B), SM-102 (8C), and iVd (8D)).

[0093] Figures 9A - 9GShows the in vivo transfection efficiency of luciferase-expressing mRNA. The LNPs were formulated with iVd lipids and 1 μg of the formulated luciferase-expressing mRNA was intramuscularly injected. After administration, (9A-9D) show representative images of luciferase expression measured by whole-body bioluminescence imaging of the draining lymph nodes (dLNs) and injection site dorsally and ventrally using an IVIS Spectrum in vivo imaging system at 6 (9A), 24 (9B), 48 (9C), and 96 (9D) hours, and (9E-9G) show graphs of quantitative luciferase expression measured by whole-body bioluminescence imaging of the injection site (9E), dLNs (9F), and liver (9G) using an IVIS Spectrum in vivo imaging system at 4, 24, 48, and 96 hours. dLN represents the draining lymph node.

[0094] The same reference signs in the respective schematics indicate the same elements. Detailed Description

[0095] The present disclosure provides novel compounds, nanomaterials, and their uses. Compositions comprising the compounds and agents of the present invention are also provided. The present disclosure also provides methods of delivering an agent to a subject using the compositions. These nanomaterials are used in applications such as gene therapy and drug delivery.

[0096] Embodiments of the present invention will now be described in detail, examples of which are shown in the accompanying drawings and examples. However, the present invention may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

[0097] 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 invention belongs. The following definitions are provided to facilitate a full understanding of the terms used in this specification.

[0098] Definitions

[0099] General Definitions

[0100] As used in this specification and the appended claims, the terms "comprising" (and its forms, derivatives or variations such as "comprises" and "comprising") and "including" (and its forms, derivatives or variations such as "includes" and "including") are inclusive (i.e., open-ended) and do not exclude additional elements or steps. For example, when used in this specification, the term "comprise and / or comprising" specifies the presence of the stated features, integers, steps, operations, elements and / or components, but does not exclude the presence or addition of one or more other features, integers, steps, operations, elements, components and / or groups thereof. Except where otherwise noted, all numbers expressing quantities of ingredients, reaction conditions, geometric shapes, dimensions, etc. used in the specification and claims should be understood at least, and are not intended to limit the application of the doctrine of equivalents to the scope of the claims, and are to be interpreted in accordance with the number of significant figures and ordinary rounding methods.

[0101] Accordingly, these terms are intended to cover not only the recited elements or steps but also other elements or steps that may not be expressly recited. In addition, as used herein, the use of the terms "a" and "the" in conjunction with an element may mean "one", but it is also consistent with the meaning of "one or more", "at least one" and "one or more than one". Thus, without further limitation, an element preceded by "a" or "an" does not exclude the presence of additional identical elements.

[0102] In this document, ranges may be expressed as from "about" a particular value and / or to "about" another particular value. "About" means within 5% of the stated value, e.g., within 4%, 3%, 2% or 1% of the stated value. When expressing such ranges, on the other hand, includes from the one particular value and / or to the other particular value. Similarly, when a value is expressed as an approximation by use of the antecedent "about", it should be understood that the particular value forms on the other hand. It should be further understood that each endpoint of each range is significant with respect to the other endpoint, and independently of the other endpoint. It should also be understood that many values are disclosed herein, and in addition to the value itself, each value is also disclosed herein as "about" the particular value. For example, if the value "10" is disclosed, then "about 10" is also disclosed. A range may be interpreted to include the beginning and end of the range. For example, a range of 10% to 20% (i.e., a range of 10% to 20%) may include 10% and also include 20%, and includes the percentages between 10% and 20%, unless otherwise expressly stated herein.

[0103] As used herein, the terms “may,” “optionally,” and “optionally may” are used interchangeably and mean including both the case where the condition occurs and the case where the condition does not occur. Thus, for example, the statement that a formulation “may include an excipient” means including both the case where the formulation includes the excipient and the case where the formulation does not include the excipient.

[0104] It should be understood that when combinations, subsets, groups, etc. of elements are disclosed (e.g., combinations of components in a composition or combinations of steps in a method), although specific references to each of the various individual and collective combinations and permutations of these elements may not be explicitly disclosed, each element is specifically contemplated and described herein.

[0105] “Administering” to a subject includes any route by which a medicament is introduced or delivered to the subject. Administration can be effected by any suitable route, including orally, topically, transdermally, transepithelially, intra-articularly, intra-arterially, intradermally, intraventricularly, intralesionally, intranasally, rectally, vaginally, by inhalation, by an implanted reservoir, parenterally (e.g., subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intraperitoneal, intrahepatic, intralesional, and intracranial injection or infusion techniques), etc. As used herein, “concurrent administration,” “combined administration,” “simultaneous administration,” or “co-administered simultaneously” means that the compounds are administered at the same point in time or substantially one immediately after the other. In the latter case, the administrations of the two compounds are close enough in time such that the observed results are indistinguishable from those obtained when the compounds are administered at the same point in time. “Systemic administration” means introducing or delivering a medicament to a subject by a route that results in the introduction or delivery of the medicament to a broad area of the subject's body (e.g., greater than 50% of the body), such as by entry into the circulatory or lymphatic systems. In contrast, “local administration” means introducing or delivering a medicament to a subject by a route that results in the introduction or delivery of the medicament to the area at or immediately adjacent to the site of administration and does not result in the introduction of the medicament systemically in a therapeutically significant amount. For example, a locally administered medicament can be readily detected locally near the site of administration but is not detectable or is detectable in negligible amounts in distal portions of the subject's body. Administration includes self-administration and administration by another person.

[0106] As used herein, the terms “controlled release” or “controlled release drug delivery” or “extended release” refer to the release or administration of a drug from a given dosage form in a controlled manner to achieve desired in vivo pharmacokinetic characteristics. One aspect of “controlled” drug delivery is the ability to manipulate the formulation and / or dosage form in order to establish the desired drug release kinetics.

[0107] As used herein, the terms "beneficial agent" and "active agent" are used interchangeably herein to refer to a chemical compound or composition having a beneficial biological effect. Beneficial biological effects include therapeutic effects, i.e., treating a disorder or other undesirable physiological condition, and prophylactic effects, i.e., preventing a disorder or other undesirable physiological condition. The terms also encompass pharmaceutically acceptable pharmacologically active derivatives of the beneficial agents specifically mentioned herein, including but not limited to salts, esters, amides, prodrugs, active metabolites, isomers, fragments, analogs, etc. When the term "beneficial agent" or "active agent" is used, or when a specific agent is specifically identified, it should be understood that the term includes the agent itself as well as pharmaceutically acceptable pharmacologically active salts, esters, amides, prodrugs, conjugates, active metabolites, isomers, fragments, analogs, etc.

[0108] A "therapeutic agent" refers to any composition having a beneficial biological effect. Beneficial biological actions include: a therapeutic action, e.g., treating a disorder or other undesirable physiological condition; and a prophylactic action, e.g., preventing a disorder or other undesirable physiological condition. The term also encompasses pharmaceutically acceptable pharmacologically active derivatives of the beneficial agents specifically mentioned herein, including but not limited to salts, esters, amides, proagents, active metabolites, isomers, fragments, analogs, etc. When the term "therapeutic agent" is used, or when a specific agent is specifically identified, it should be understood that the term includes the agent itself as well as pharmaceutically acceptable pharmacologically active salts, esters, amides, proagents, conjugates, active metabolites, isomers, fragments, analogs, etc.

[0109] "Reduce" can refer to any change that results in a lesser amount of a symptom, disease, composition, condition, or activity. A substance should also be understood to reduce the genetic output of a gene when the genetic output of the gene product of the substance is lower relative to the output without the gene product of the substance. Also, for example, a reduction can be a change in the symptoms of a disorder such that the symptoms are less than previously observed. A reduction can be any individual, median, or average reduction of a condition, symptom, activity, composition in a statistically significant amount. Thus, a reduction can be a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% reduction, provided the reduction is statistically significant.

[0110] "Inhibit", "inhibiting", and "inhibition" mean a decrease in activity, response, condition, disease, or other biological parameter. This can include, but is not limited to, complete ablation of activity, response, condition, or disease. This can also include, for example, a 10% decrease in activity, response, condition, or disease compared to a native or control level. Thus, the decrease can be 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any amount in between compared to a native or control level.

[0111] "Inactivate", "inactivating", and "inactivation" refer to the reduction or elimination of activity, response, condition, disease, or other biological parameter due to a chemical (covalent bond formation) between a ligand and its biological target.

[0112] "Reduce" or other forms of the word, such as "reducing" or "reduction", mean a decrease in an event or property (e.g., tumor growth). It should be understood that this is usually relative to a standard or expected value, in other words it is relative, but it is not always necessary to refer to a standard or relative value. For example, "reduce tumor growth" means a decrease in the rate of tumor growth relative to a standard or control.

[0113] As used herein, the terms "treating" or "treatment" of a subject include administering a medicament to a subject for the purpose of preventing, curing, healing, alleviating, ameliorating, altering, remedying, improving, enhancing, stabilizing, or affecting a disease or disorder, or the symptoms of a disease or disorder. The terms "treating" and "treatment" can also refer to reducing the severity and / or frequency of symptoms, eliminating symptoms and / or their root cause, preventing the occurrence of symptoms and / or their root cause, and improving or remedying impairment.

[0114] "Prevent" or other forms of the word such as "preventing" or "prevention" mean to stop a particular event or characteristic, to stabilize or delay the development or progression of a particular event or characteristic, or to minimize the chance of a particular event or characteristic occurring. Prevention does not require comparison to a control because it is generally more absolute than, for example, reduction. As used herein, something can reduce but not prevent, but something that reduces can also prevent. Similarly, something can prevent but not reduce, but something that prevents can also reduce. It should be understood that unless otherwise expressly stated, the use of other words is also expressly disclosed when reduction or prevention is used. For example, the terms "prevent" or "inhibit" can refer to a treatment that prevents or slows the onset of a disease or condition or reduces the severity of a disease or condition in advance. Thus, if a treatment can treat a disease in a subject having symptoms of the disease, it can also prevent or inhibit the disease in a subject who has not yet suffered some or all of the symptoms. As used herein, the term "prevent" a disorder or adverse physiological event in a subject specifically refers to preventing the occurrence of symptoms and / or its underlying cause, where the subject may or may not exhibit a high sensitivity to the disorder or event.

[0115] The term "effective amount" of a therapeutic agent means an amount of a beneficial agent that is non-toxic but sufficient to provide the desired effect. The amount of a "effective" beneficial agent will vary from subject to subject, depending on the age and general condition of the subject, the particular beneficial agent or agents, etc. Thus, it is not always possible to specify an exact "effective amount". However, one of ordinary skill in the art can determine the appropriate "effective amount" in any subject case using routine experimentation. Moreover, as used herein and unless otherwise expressly stated, the "effective amount" of a beneficial agent can also refer to an amount that encompasses both a therapeutically effective amount and a prophylactically effective amount.

[0116] The "effective amount" of a drug necessary to achieve a therapeutic effect can vary depending on factors such as the age, sex, and weight of the subject. The dosage regimen can be adjusted to provide an optimal therapeutic response. For example, several separate doses can be administered daily, or the dosage can be proportionally reduced as indicated by the exigencies of the treatment situation.

[0117] As used herein, a "therapeutically effective amount" of a therapeutic agent refers to an amount effective to achieve a desired therapeutic outcome, and a "prophylactically effective amount" of a therapeutic agent refers to an amount effective to prevent an adverse physiological condition. The therapeutically effective amount and prophylactically effective amount of a given therapeutic agent will generally vary depending on factors such as the type and severity of the condition or disease being treated and the age, sex, and weight of the subject. The term "therapeutically effective amount" can also refer to the amount of a therapeutic agent or the rate of delivery of a therapeutic agent (e.g., an amount that varies over time) that effectively promotes a desired therapeutic effect. The precise desired therapeutic effect will vary depending on the condition to be treated, the tolerance of the subject, the drug and / or pharmaceutical formulation to be administered (e.g., the potency of the therapeutic agent (drug), the concentration of the drug in the formulation, etc.), and various other factors understood by one of ordinary skill in the art.

[0118] As used herein, the term "pharmaceutically acceptable" component can refer to a component that is not biologically or otherwise undesirable, i.e., the component can be incorporated into a pharmaceutical formulation of the invention and administered to a subject as described herein without causing any significant undesirable biological effects or interacting in a harmful manner with any other components of the formulation containing it. When the term "pharmaceutically acceptable" is used to refer to an excipient, the term generally means that the component has met the required standards of toxicological and manufacturing tests or that the component is included in the Inactive Ingredient Guide established by the U.S. Food and Drug Administration.

[0119] A "pharmaceutically acceptable carrier" (sometimes referred to as a "carrier") means a carrier or excipient that can be used to prepare a pharmaceutical or therapeutic composition that is generally safe and non-toxic and includes carriers acceptable for veterinary and / or human pharmaceutical or therapeutic use. The term "carrier" or "pharmaceutically acceptable carrier" can include, but is not limited to, phosphate buffered saline solutions, water, emulsions (such as oil / water or water / oil emulsions), and / or various types of wetting agents. As used herein, the term "carrier" encompasses, but is not limited to, any excipient, diluent, filler, salt, buffer, stabilizer, solubilizer, lipid, stabilizer, or other materials known in the art for use in pharmaceutical formulations and further described herein.

[0120] As used herein, "pharmaceutically acceptable salts" are derivatives of the disclosed compounds, wherein the parent compound is modified by making its inorganic and organic, non-toxic acid or base addition salts. The salts of the compounds of the present invention can be synthesized from the parent compounds containing basic or acidic moieties by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid forms of these compounds with a stoichiometric amount of a suitable base (such as Na, Ca, Mg or K hydroxides, carbonates, bicarbonates, etc.) or by reacting the free base forms of these compounds with a stoichiometric amount of a suitable acid. Such reactions are usually carried out in water or an organic solvent or a mixture of both. Generally, when feasible, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are typical. The salts of the compounds of the present invention further include solvates of the compounds and compound salts.

[0121] Examples of pharmaceutically acceptable salts include, but are not limited to: mineral salts or organic salts of basic residues such as amines; base salts or organic salts of acidic residues such as carboxylic acids; and the like. Pharmaceutically acceptable salts include, for example, conventional non-toxic salts and quaternary ammonium salts of the parent compounds formed from non-toxic inorganic or organic acids. For example, conventional non-toxic acid salts include salts derived from the following inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, etc.; and salts prepared from the following organic acids: acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, hydroxyethanesulfonic acid, HOOC-(CH2)n-COOH (where n is 0-4), etc., or salts prepared using different acids that produce the same counterion. A list of additional suitable salts can be found, for example, in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, Pa., page 1418 (1985).

[0122] In addition, as used herein, the term "pharmacologically active" (or simply "active") in "pharmacologically active" derivatives or analogs can refer to derivatives or analogs (such as salts, esters, amides, conjugates, metabolites, isomers, fragments, etc.) that have the same type of pharmacological activity as the parent compound and to a substantially equal degree.

[0123] A "control" is an alternative subject or sample used for comparison purposes in an experiment. A control can be "positive" or "negative".

[0124] As used herein, "subject" means an individual. Thus, "subject" can include domestic animals (e.g., cats, dogs, etc.), livestock (e.g., cows, horses, pigs, sheep, goats, etc.), laboratory animals (e.g., mice, rabbits, rats, guinea pigs, goats, sheep, pigs, dogs, cats, etc.), and birds (e.g., chickens, turkeys, songbirds, etc.). "Subject" can also include mammals such as primates or humans. Thus, a subject can be a human or a veterinary patient. The term "patient" refers to a subject under the treatment of a clinician, such as a physician. Administration of a therapeutic agent can be carried out at a dose and for a duration effective to treat the subject. In some embodiments, the subject is a human.

[0125] As used herein, the term "nucleic acid" refers to a polymer composed of nucleotides, such as deoxyribonucleotides or ribonucleotides.

[0126] As used herein, the terms "ribonucleic acid" and "RNA" refer to a polymer composed of ribonucleotides.

[0127] As used herein, the terms "deoxyribonucleic acid" and "DNA" refer to a polymer composed of deoxyribonucleotides.

[0128] The term "oligonucleotide" refers to a single-stranded or double-stranded nucleotide polymer having a length of from about 2 to up to about 100 nucleotides. Suitable oligonucleotides can be prepared by the phosphoramidite method described by Beaucage and Carruthers, Tetrahedron Lett., 22:1859-1862 (1981) or by the triester method according to Matteucci, et al., J. Am. Chem. Soc., 103:3185 (1981), both of which are incorporated herein by reference, or by other chemical methods using commercial automated oligonucleotide synthesizers or VLSIPS TM techniques. When an oligonucleotide is referred to as "double-stranded", one of ordinary skill in the art will understand that a pair of oligonucleotides exists in a helical array of hydrogen bonds typically associated with, for example, DNA. In addition to the 100% complementary form of double-stranded oligonucleotides, the term "double-stranded" as used herein also means those forms involving such structural features as bulges and loops, which are more fully described in such biochemical texts as Stryer, Biochemistry, Third Edition, (1988), which is incorporated herein by reference for all purposes.

[0129] The term "polynucleotide" refers to a single-stranded or double-stranded polymer composed of nucleotide monomers. In some embodiments, the polynucleotide is composed of nucleotide monomers having a length generally greater than 100 nucleotides and a length of up to about 8,000 or more nucleotides.

[0130] The term "polypeptide" refers to a compound composed of a single chain of D- or L-amino acids or a mixture of D- and L-amino acids joined by peptide bonds.

[0131] The term "complementary" refers to the topological compatibility or fitting together of the interaction surfaces of a probe molecule and its target. Thus, the target and its probe can be described as complementary, and further, the contacting surface characteristics are complementary to each other.

[0132] The term "recombinant" refers to a nucleic acid (e.g., polynucleotide) that has been manipulated by a human or a copy or complement of a nucleic acid (e.g., polynucleotide) that has been manipulated by a human, or, if referring to a protein (i.e., "recombinant protein"), a protein encoded by a recombinant nucleic acid (e.g., polynucleotide). In an embodiment, a recombinant expression cassette comprising a promoter operably linked to a second nucleic acid (e.g., polynucleotide) can include a promoter that is heterologous to the second nucleic acid (e.g., polynucleotide) as a result of human manipulation (e.g., by the methods described in Sambrook et al., "Molecular Cloning - A Laboratory Manual", Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. (1989) or "Current Protocols in Molecular Biology", Volumes 1-3, John Wiley & Sons, Inc. (1994-1998)). In another example, a recombinant expression cassette can contain nucleic acids (e.g., polynucleotides) combined in a manner that is highly unlikely to be found in nature. For example, a human-engineered restriction site or plasmid vector sequence can flank the promoter or separate it from the second nucleic acid (e.g., polynucleotide). Those skilled in the art will recognize that nucleic acids (e.g., polynucleotides) can be manipulated in a variety of ways and are not limited to the above examples.

[0133] In the context of two or more nucleic acid or polypeptide sequences, the terms "identical" or "identity" percent refers to two or more sequences or subsequences that have a specified percentage of identical or same amino acid residues or nucleotides when compared and aligned for maximum correspondence over a comparison window or specified region (i.e., about 60% identity over a particular region, preferably 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or higher identity), using the BLAST or BLAST 2.0 sequence comparison algorithms with the default parameters described below or by manual alignment and visual inspection (see, e.g., the NCBI web site, etc.). Such sequences are then said to be "substantially identical". This definition also relates to or can be applied to the complement of a test sequence. The definition also includes sequences with deletions and / or additions, as well as sequences with substitutions. As described below, preferred algorithms can account for gaps, etc. Preferably, the identity exists over a region of at least about 10 amino acids or 20 nucleotides in length, or more preferably over a region of 10 to 50 amino acids or 20 to 50 nucleotides in length. As used herein, the percent amino acid sequence identity (%) is defined as the percentage of amino acids in a candidate sequence that are identical to the amino acids in a reference sequence after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percent sequence identity. The alignment for determining the percent sequence identity can be achieved in various ways within the skill in the art, e.g., using publicly available computer software such as BLAST, BLAST-2, ALIGN, ALIGN-2, or Megalign (DNASTAR) software. The appropriate parameters for measuring the alignment, including any algorithms needed to achieve the maximum alignment over the full length of the sequences being compared, can be determined by known methods.

[0134] For sequence comparison, typically one sequence acts as a reference sequence to which a test sequence is compared. When using a sequence comparison algorithm, the test and reference sequences are input into a computer, subsequence coordinates are designated (if necessary), and sequence algorithm program parameters are designated. Preferably, the default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identity of the test sequence relative to the reference sequence based on the program parameters.

[0135] An example of an algorithm suitable for determining percent sequence identity and percent sequence similarity are the BLAST and BLAST 2.0 algorithms, described in Altschul et al. (1977) Nuc. Acids Res. 25:3389-3402, and Altschul et al. (1990) J. Mol. Biol. 215:403-410, respectively. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov / ). The algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence that either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al. (1990) J. Mol. Biol. 215:403-410). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing the initial neighborhood word hits. Word hits are extended in both directions along each sequence until the cumulative alignment score can no longer increase. For nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for a mismatch residue; always <0) are used to calculate the cumulative score. For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction stops when: the cumulative alignment score falls by the quantity X from its maximum achieved value; the cumulative score goes to zero or below due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, M = 5, N = -4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff (1989) Proc. Natl. Acad. Sci. USA 89:10915), an alignment (B) of 50, an expectation (E) of 10, M = 5, N = -4, and a comparison of both strands.

[0136] The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5787). One measure of similarity provided by the BLAST algorithm is the minimum sum probability (P(N)), which provides an indication of the probability that a match occurs by chance between two nucleotide or amino acid sequences. For example, if the minimum sum probability is less than about 0.2, more preferably less than about 0.01, in a comparison of a test nucleic acid with a reference nucleic acid, the nucleic acid is considered to be similar to the reference sequence.

[0137] The term "nucleobase" refers to the portion of a nucleotide that has Watson / Crick base-pairing functionality. The most common natural nucleobases, adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T), have hydrogen-bonding functionality to bind one nucleic acid strand to another in a sequence-specific manner.

[0138] A nucleic acid sequence is "heterologous" to a second nucleic acid sequence when it is derived from a foreign species or, when from the same species, is modified from its original form by human activity. For example, a heterologous promoter (or heterologous 5' untranslated region (5'UTR)) operably linked to a coding sequence refers to a coding sequence from a species different from the species from which the promoter is derived or, if from the same species, to a coding sequence that is different from the naturally occurring allelic variant (e.g., a 5'UTR or 3'UTR from a different gene is operably linked to a nucleic acid encoding a costimulatory molecule).

[0139] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies within the population are identical except for possible naturally occurring mutations that may be present in a minor subset of the antibody molecules. Monoclonal antibodies herein specifically include "chimeric" antibodies in which a portion of the heavy and / or light chain is identical or homologous to the corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical or homologous to the corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass and fragments of such antibodies, so long as they exhibit the desired antagonistic activity.

[0140] The disclosed monoclonal antibodies can be prepared using any procedure for producing monoclonal antibodies. For example, the disclosed monoclonal antibodies can be prepared using the hybridoma method, such as those described by Kohler and Milstein, Nature, 256:495 (1975). In the hybridoma method, a mouse or other suitable host animal is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent. Alternatively, lymphocytes can be immunized in vitro.

[0141] Monoclonal antibodies can also be prepared by recombinant DNA methods. The DNA encoding the disclosed monoclonal antibodies can be readily isolated and sequenced using conventional methods (e.g., by using oligonucleotide probes that specifically bind to the genes encoding the heavy and light chains of murine antibodies). Phage display techniques (e.g., as described in U.S. Patent No. 5,804,440 to Burton et al. and U.S. Patent No. 6,096,441 to Barbas et al.) can also be used to generate and screen libraries of antibodies or active antibody fragments.

[0142] In vitro methods are also suitable for preparing monovalent antibodies. Digestion of an antibody to produce its fragments, particularly Fab fragments, can be accomplished using conventional techniques known in the art. For example, digestion can be carried out with papain. Examples of papain digestion are described in WO 94 / 29348 published on December 22, 1994 and U.S. Patent No. 4,342,566. Papain digestion of an antibody generally produces: two identical antigen-binding fragments, called Fab fragments, each of which has a single antigen-binding site; and a residual Fc fragment. Pepsin treatment yields fragments that have two antigen-combining sites and are still capable of cross-linking antigens.

[0143] As used herein, the terms “antibody or its antigen-binding fragment” or “antibody or its fragment” encompass chimeric and hybrid antibodies having dual or multiple antigen or epitope specificities, as well as fragments such as F(ab')2, Fab', Fab, Fv, sFv, scFv, etc., including hybrid fragments. Thus, antibody fragments that retain the ability to bind the specific antigen of the antibody are provided. For example, antibody fragments that maintain binding activity are included within the meaning of the term “antibody or its antigen-binding fragment”. Such antibodies and fragments can be prepared by techniques known in the art and can be screened for specificity and activity according to the examples and the methods set forth in the general methods for generating and screening antibodies for specificity and activity (see Harlow and Lane, Antibodies, A Laboratory Manual, Cold Spring Harbor Publications, New York, (1988)).

[0144] The meaning of “antibody or its antigen-binding fragment” also includes conjugates of antibody fragments and antigen-binding proteins (single-chain antibodies). The meaning of “antibody or its antigen-binding fragment” also includes immunoglobulin single variable domains, such as, for example, nanobodies.

[0145] Fragments, whether or not attached to other sequences, may also include insertions, deletions, substitutions or other selected modifications of specific regions or specific amino acid residues, provided that the activity of the antibody or antibody fragment is not significantly changed or impaired compared to the unmodified antibody or antibody fragment. These modifications may provide some additional properties, such as removal / addition of amino acids capable of disulfide bonding, to increase their biological lifespan, change their secretion properties, etc. In any case, the antibody or antibody fragment must have biologically active properties, such as specific binding to its cognate antigen. The function or active region of an antibody or antibody fragment may be identified by mutagenesis of a specific region of the protein, subsequent expression and testing of the expressed polypeptide. Such methods are obvious to those skilled in the art and may include site-specific mutagenesis of nucleic acids encoding antibodies or antibody fragments. (Zoller, MJ "New Views on Biotechnology (Curr. Opin. Biotechnol.)" 3:348-354, 1992).

[0146] As used herein, the term "antibody" or "antibodies" may also refer to human antibodies and / or humanized antibodies. Many non-human antibodies (e.g., antibodies derived from mice, rats, or rabbits) have natural antigenicity in humans and therefore may cause an undesirable immune response when administered to humans. Therefore, human or humanized antibodies are used in the method to reduce the chance that antibodies administered to humans cause an undesirable immune response.

[0147] Chemical Definition

[0148] As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, and aromatic and non-aromatic substituents of organic compounds. Illustrative substituents include, for example, those described below. For appropriate organic compounds, permissible substituents may be one or more and may be the same or different. For the purposes of this disclosure, heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of heteroatoms. This disclosure is not intended to be limited in any way to permissible substituents of organic compounds. Furthermore, the term "substituted" or "substituted by..." includes the following implicit prerequisites: such substitutions are consistent with the permissible valences of the substituted atoms and substituents, and the substitution produces a stable compound, such as a compound that will not spontaneously transform as by rearrangement, cyclization, elimination, etc.

[0149] "Z 1 ”, “Z 2 ”, “Z 3 ” and “Z 4” is used herein as a general symbol representing various specific substituents. These symbols can be any substituent, not limited to those disclosed herein, and when they are defined as certain substituents in one case, they can be defined as some other substituents in another case.

[0150] As used herein, the term "aliphatic" refers to non-aromatic hydrocarbon groups and includes branched and unbranched alkyl, alkenyl or alkynyl groups.

[0151] As used herein, the term "alkyl" is a branched or unbranched saturated hydrocarbon group of 1 to 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, tetradecyl, hexadecyl, eicosyl, tetracosyl, etc. The alkyl can also be substituted or unsubstituted. The alkyl group can be substituted by one or more groups, including but not limited to alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide or thiol, as described below.

[0152] Throughout this specification, "alkyl" is generally used to refer to both unsubstituted alkyl and substituted alkyl; however, the substituted alkyl is also specifically mentioned herein by identifying one or more specific substituents on the alkyl. For example, the term "haloalkyl" specifically refers to an alkyl substituted by one or more halides, such as fluorine, chlorine, bromine or iodine. The term "alkoxyalkyl" specifically refers to an alkyl substituted by one or more alkoxy groups, as described below. The term "alkylamino" specifically refers to an alkyl substituted by one or more amino groups, etc., as described below. When "alkyl" is used in one case and a specific term such as "alkyl alcohol" is used in another case, this does not imply that the term "alkyl" also does not refer to specific terms such as "alkyl alcohol".

[0153] This practice is also used for other groups described herein. That is, although terms such as "cycloalkyl" refer to both unsubstituted and substituted cycloalkyl moieties, the substituted moiety can also be specifically identified herein; for example, a specifically substituted cycloalkyl can be referred to as, for example, "alkylcycloalkyl". Similarly, a substituted alkoxy can be specifically referred to as, for example, "haloalkoxy", and a specifically substituted alkenyl can be, for example, "alkenyl alcohol", etc. Again, the practice of using a general term such as "cycloalkyl" and a specific term such as "alkylcycloalkyl" does not imply that the general term does not also include the specific term.

[0154] As used herein, the term "alkoxy" refers to an alkyl bonded through a single terminal ether bond; that is, the "alkoxy" group can be defined as —OZ 1, where Z 1 is the alkyl group defined above.

[0155] As used herein, the term "alkenyl" refers to a hydrocarbyl group having 2 to 24 carbon atoms and having at least one carbon-carbon double bond in its structural formula. As in (Z 1 Z 2 )C═C(Z 3 Z 4 ) and other asymmetric structures are intended to include both E and Z isomers. This may be assumed in the structural formulas herein where there are asymmetric alkenes, or it may be explicitly indicated by the bond symbol C═C. The alkenyl group may be substituted by one or more groups, including but not limited to alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide or thiol as described hereinbelow.

[0156] As used herein, the term "alkynyl" refers to a hydrocarbyl group having 2 to 24 carbon atoms and having at least one carbon-carbon triple bond in its structural formula. The alkynyl group may be substituted by one or more groups, including but not limited to alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfo-oxy, sulfonyl, sulfone, sulfoxide or thiol as described hereinbelow.

[0157] As used herein, the term "aryl" refers to a group containing any carbon-based aromatic group, including but not limited to benzene, naphthalene, phenyl, biphenyl, phenoxyphenyl, etc. The term "heteroaryl" is defined as a group containing an aromatic group having at least one heteroatom incorporated within the ring. Examples of heteroatoms include but are not limited to nitrogen, oxygen, sulfur and phosphorus. The term "non-heteroaryl" included within the term "aryl" defines a group containing an aromatic group that does not contain a heteroatom. The aryl or heteroaryl may be substituted or unsubstituted. The aryl or heteroaryl group may be substituted by one or more groups, including but not limited to alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide or thiol as described herein. The term "biaryl" is a specific type of aryl and is included within the definition of "aryl". A biaryl refers to two aryl groups joined together by a fused ring structure as in naphthalene or by one or more carbon-carbon bonds as in biphenyl.

[0158] As used herein, the term "cycloalkyl" is a non-aromatic carbon-based ring composed of at least three carbon atoms. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. The term "heterocycloalkyl" is a cycloalkyl as defined above in which at least one of the carbon atoms in the ring is replaced by a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. Cycloalkyl and heterocycloalkyl may be substituted or unsubstituted. The cycloalkyl group and the heterocycloalkyl group may be substituted by one or more groups including, but not limited to, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide, or thiol as described herein.

[0159] As used herein, the term "cycloalkenyl" is a non-aromatic carbon-based ring composed of at least three carbon atoms and containing at least one double bond, i.e., C═C. Examples of cycloalkenyl include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and the like. The term "heterocycloalkenyl" is a cycloalkenyl group as defined above and included within the meaning of the term "cycloalkenyl" in which at least one of the carbon atoms in the ring is replaced by a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus. Cycloalkenyl and heterocycloalkenyl may be substituted or unsubstituted. The cycloalkenyl group and the heterocycloalkenyl group may be substituted by one or more groups including, but not limited to, alkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfo-oxo, sulfonyl, sulfone, sulfoxide, or thiol as described herein.

[0160] The term "cyclic group" is used herein to refer to an aryl group, a non-aryl group (i.e., cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl groups), or both. A cyclic group has one or more ring systems that may be substituted or unsubstituted. A cyclic group may contain one or more aryl groups, one or more non-aryl groups, or one or more aryl groups and one or more non-aryl groups.

[0161] The term "aldehyde" as used herein is represented by the formula -C(O)H. Throughout the specification, "C(O)" or "CO" is a shorthand notation for C═O.

[0162] "Amine" or "amino" as used herein is represented by the formula —NZ 1 Z 2 wherein Z 1 and Z 2 may each be a substituted group as described above such as hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl, or heterocycloalkenyl group.

[0163] As used herein, the term "carboxylic acid" is represented by the formula -C(O)OH. As used herein, a "carboxylate" or "carboxyl" group is represented by the formula —C(O)O - is represented by

[0164] As used herein, the term "ester" is represented by the formula —OC(O)Z 1 or —C(O)OZ 1 is represented, where Z 1 can be an alkyl, haloalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl group as described above.

[0165] As used herein, the term "ether" is represented by the formula Z 1 OZ 2 is represented, where Z 1 and Z 2 can independently be an alkyl, haloalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl group as described above.

[0166] As used herein, the term "ketone" is represented by the formula Z 1 C(O)Z 2 is represented, where Z 1 and Z 2 can independently be an alkyl, haloalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl group as described above.

[0167] As used herein, the terms "halide" or "halogen" refer to fluorine, chlorine, bromine and iodine.

[0168] As used herein, the term "hydroxyl" is represented by the formula -OH.

[0169] As used herein, the term "nitro" is represented by the formula -NO2.

[0170] As used herein, the term "silyl" is represented by the formula —SiZ 1 Z 2 Z 3 is represented, where Z 1 、Z 2 and Z 3 can independently be hydrogen, an alkyl, haloalkyl, alkoxy, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl group as described above.

[0171] The term "sulfonyl" is used herein to refer to a sulfo-oxo group represented by the formula —S(O)2Z 1 is represented, where Z 1It may be a hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl group as described above.

[0172] As used herein, the term "sulfonylamino" or "benzenesulfonamide" is represented by the formula —S(O)2NH—.

[0173] The term "phosphonyl" as used herein refers to a phospho-oxo group represented by the formula —P(O)(OZ 1 )2, where Z 1 may be a hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocycloalkyl or heterocycloalkenyl group as described above.

[0174] As used herein, the term "thiol" is represented by the formula -SH.

[0175] The term "thio" as used herein is represented by the formula —S—.

[0176] As used herein, "R 1 ", "R 2 ", "R 3 ", "R n ", etc. (where n is an integer) may independently have one or more of the groups listed above. For example, if R 1 is a straight-chain alkyl group, one of the hydrogen atoms of the alkyl group may optionally be substituted with a hydroxyl group, an alkoxy group, an amino group, an alkyl group, a halide, etc. Depending on the groups selected, the first group may be incorporated within the second group, or alternatively, the first group may be a side group of the second group (i.e., connected to the second group). For example, for the phrase "alkyl group containing an amino group", the amino group may be incorporated within the main chain of the alkyl group. Alternatively, the amino group may be connected to the main chain of the alkyl group. The nature of the groups selected will determine whether the first group is embedded within the second group or connected to the second group.

[0177] As used herein, a "cationic lipid" refers to a lipid having a group or atom that carries a positive charge at any pH value or hydrogen ion activity in its environment. Typically, the positive charge is due to the presence of a quaternary nitrogen atom. When a compound carries multiple such positive charges, it may be referred to as a polycationic lipid. Thus, the cationic lipid according to one embodiment of the present invention is positively charged at any pH value of its environment, including under physiological conditions, such as under the physiological salt conditions of cells in vivo. In some embodiments, the cationic lipid may include a group or atom that is positively charged at any pH value or hydrogen ion activity of its environment and a group or atom that is charged according to the environmental pH (e.g., a cationic group and an ionizable group).

[0178] As used herein, the term "ionizable lipid" refers to a lipid having a group or atom that is charged (ionized) depending on the environmental pH. An ionizable lipid can have at least one protonatable or deprotonatable group such that the lipid is positively charged at a pH equal to or below physiological pH (e.g., pH 7.4) and neutral at a second pH, which second pH is preferably equal to or above physiological pH. Generally, the charge of primary, secondary, and tertiary amines depends on the pH of the system. At low pH levels, these amines tend to be strongly cationic. At high pH levels, these amines do not ionize. One of ordinary skill in the art will understand that the addition or removal of protons as a function of pH is an equilibrium process, and the reference to charged or neutral lipids refers to the nature of the predominant species, without requiring that all of the lipids be present in the charged or neutral form.

[0179] As used herein, the term "vitamin moiety" refers to a portion of a compound that is structurally similar to a vitamin, such as a portion derived by covalently linking (or being covalently linkable) a vitamin to a precursor of a compound having a vitamin moiety. In this context, the term "vitamin" refers to an organic compound that is essential as an important nutrient for a subject but that the subject cannot synthesize in sufficient amounts. Exemplary vitamins include, but are not limited to, vitamin A, vitamin B (e.g., vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B12), vitamin C, vitamin D (e.g., vitamin D3), vitamin E, vitamin K (e.g., vitamin K1, vitamin K2), biotin, folate, folic acid, lipoic acid, ascorbic acid, choline, carnitine, carotenoids (e.g., β-carotene, α-carotene, γ-carotene), and salts and derivatives thereof. The term vitamin also includes derivatives of vitamins and molecules or products related to vitamin synthesis, absorption, or processing, such as thiamine pyrophosphate (TPP), flavin mononucleotide (FMM), flavin adenine dinucleotide (FAD), nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide phosphate (NADP), coenzyme-A (CoA), pyridoxal phosphate, biocytin, tetrahydrofolate, coenzyme B12, lipoyllysine, 1,1-cis-retinal, and 1,2,5-dihydroxycholecalciferol.

[0180] Unless stated to the contrary, a formula having chemical bonds shown only as solid lines rather than as wedges or dashes contemplates every possible isomer, such as each enantiomer, diastereomer, and meso compound, as well as mixtures of isomers, such as a racemic or scalemic mixture.

[0181] Reference will now be made in detail to specific aspects of the disclosed materials, compounds, compositions, articles, and methods, examples of which are illustrated in the accompanying examples and figures.

[0182] Compound

[0183] Compounds of formula Da, Db, or Dc are described herein:

[0184]

[0185] wherein:

[0186] Z is a vitamin;

[0187] X is O or S;

[0188] L 1 is -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-;

[0189] L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -NR a C(=O)- or -C(=O)R a N-;

[0190] L 3 is absent, is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-;

[0191] R a is where n is an integer from 0 to 12;

[0192] x is 0, 1, or 2;

[0193] M is N, CH,

[0194] D is N or

[0195] G 1 、G 2 、G 3 and G 5Each independently selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0196] G 4 is absent or selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0197] R 2 is -CR b R c 、C6-C 24 alkyl or C6-C 24 alkenyl;

[0198] R 3 is

[0199] R 6 is H, OR b 、CN, -C(=O)OR b 、-NC(=O)R b 、-C(=O)NR b 、

[0200] R b and R c are independently H, C1-C 12 alkyl or C1-C 12 alkenyl; and

[0201] p, q, and r are independently 0, 1, 2, 3, or 4;

[0202] or a pharmaceutically acceptable salt thereof.

[0203] In some embodiments, the vitamin moiety can be selected from:

[0204]

[0205]

[0206] wherein

[0207] R 1 、R 4 and R 10 are each independently hydrogen or C1-C6 alkyl; and represents a carbon-carbon bond or a carbon-carbon double bond.

[0208] In some embodiments, the vitamin moiety can be:

[0209]

[0210] wherein

[0211] R 1 and R 4 are each independently hydrogen or C1-C6 alkyl; and represents a carbon-carbon single bond or a carbon-carbon double bond.

[0212] In some embodiments, the vitamin moiety may be selected from:

[0213]

[0214] wherein

[0215] R 10 is hydrogen or C1-C6 alkyl; and represents a carbon-carbon single bond or a carbon-carbon double bond.

[0216] In some embodiments, the vitamin moiety may be selected from:

[0217]

[0218] In some embodiments, compounds of formula Y, formula B or formula C are described:

[0219]

[0220] wherein:

[0221] R 1 , R 4 and R 10 are each independently hydrogen or C1-C6 alkyl;

[0222] represents a carbon-carbon single bond or a carbon-carbon double bond;

[0223] A is

[0224]

[0225] X is O or S;

[0226] L 1 is -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-;

[0227] L 2-O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -NR a C(=O)- or -C(=O)R a N-;

[0228] L 3 Absent, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-;

[0229] R a is where n is an integer from 0 to 12;

[0230] x is 0, 1 or 2;

[0231] M is N, CH,

[0232] D is N or

[0233] G 1 , G 2 , G 3 and G 5 each independently selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0234] G 4 is absent or selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0235] R 2 is -CR b R c , C6-C 24 alkyl or C6-C 24 alkenyl;

[0236] R 3 is

[0237] R 6 is H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b ,

[0238]

[0239] R b and R c are independently H, C1-C 12 alkyl or C1-C 12 alkenyl; and

[0240] p, q, and r are independently 0, 1, 2, 3, or 4;

[0241] or a pharmaceutically acceptable salt thereof.

[0242] In some embodiments, the compounds of formula Y, B, or C can be selected from:

[0243]

[0244] or a pharmaceutically acceptable salt thereof or any combination thereof;

[0245] wherein A is the same as formula Y, formula B, or formula C.

[0246] In some embodiments, compounds of formula I, II, III, IV, V, VI, VII, VIII, or IX are described:

[0247]

[0248]

[0249]

[0250] wherein:

[0251] R 1 , R 4 and R 10 are each independently hydrogen or C1-C6 alkyl;

[0252] represents a carbon-carbon bond or a carbon-carbon double bond;

[0253] X is O or S;

[0254] L 1 is -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-;

[0255] L 2-O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -NR a C(=O)- or -C(=O)R a N-;

[0256] L 3 absent, -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-;

[0257] R a is wherein n is an integer from 0 to 12;

[0258] x is 0, 1 or 2;

[0259] M is N, CH,

[0260] D is N or

[0261] G 1 , G 2 , G 3 and G 5 each independently selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0262] G 4 is absent or selected from C1-C 12 alkyl or C1-C 12 alkenyl;

[0263] R 2 is -CR b R c , C6-C 24 alkyl or C6-C 24 alkenyl;

[0264] R 3 is

[0265] R 6 is H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b ,

[0266] R b and R c are independently H, C1-C 12 alkyl or C1-C 12 alkenyl; and

[0267] p, q, and r are independently 0, 1, 2, 3, or 4;

[0268] or a pharmaceutically acceptable salt thereof.

[0269] In some embodiments, A is

[0270]

[0271] In some embodiments, A is

[0272]

[0273] In some embodiments, A is

[0274]

[0275] In some embodiments, R 1 is C1-C6 alkyl. In some embodiments, R 1 is H. In some embodiments, R 1 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, or hexyl. In some embodiments, R 1 is methyl. In some embodiments, R 4 is C1-C6 alkyl. In some embodiments, R 4 is H. In some embodiments, R 4 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, or hexyl. In some embodiments, R 4 is methyl. In some embodiments, R 10 is C1-C6 alkyl. In some embodiments, R 10 is H. In some embodiments, R 10 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, or hexyl. In some embodiments, R 10 is methyl.

[0276] In some embodiments, X is O. In some embodiments, X is S.

[0277] In some embodiments, R 3 is

[0278] In some embodiments, R 3 is

[0279] In some embodiments, R 6 is H, OR b , CN, -C(=O)OR b , -NC(=O)R b or -C(=O)NR b .

[0280] In some embodiments, R 6 is

[0281] In some embodiments, R 6 is

[0282] In some embodiments, R 6

[0283] In some embodiments, L 1 is -C(=O)-. In some embodiments, L 2 is -(C=O)O-. In some embodiments, L 3 is absent.

[0284] In some embodiments, G 1 is C1-C 12 alkyl. In some embodiments, G 1 is C1-C 12 alkenyl. In some embodiments, G 1 is C1-C6 alkyl. In some embodiments, G 1 is C5 alkyl. In some embodiments, G 1 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl or hexyl. In some embodiments, G 1 is pentyl.

[0285] In some embodiments, G 2 is C1-C 12 alkyl. In some embodiments, G 2 is C1-C 12 alkenyl. In some embodiments, G 2 is C1-C6 alkyl. In some embodiments, G 2 is C6 alkyl. In some embodiments, G 2 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl or hexyl. In some embodiments, G 2 is hexyl.

[0286] In some embodiments, G 3 is a C1-C 12 alkyl group. In some embodiments, G 3 is a C1-C 12 alkenyl group. In some embodiments, G 3 is a C1-C6 alkyl group. In some embodiments, G3 is a C2 alkyl group. In some embodiments, G 3 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl or hexyl. In some embodiments, G 3 is ethyl.

[0287] In some embodiments, G 4 is a C1-C 12 alkyl group. In some embodiments, G 4 is a C1-C6 alkyl group. In some embodiments, G 4 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl or hexyl. In some embodiments, G 4 is a C1-C 12 alkenyl group. In some embodiments, G 4 is absent.

[0288] In some embodiments, G 5 is a C1-C 12 alkyl group. In some embodiments, G 5 is a C1-C 12 alkenyl group. In some embodiments, G 5 is a C1-C6 alkyl group. In some embodiments, G 5 is a C2 alkyl group. In some embodiments, G 5 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl or hexyl. In some embodiments, G 5 is methyl.

[0289] In some embodiments, R 2 is -CR b R c In some embodiments, R b and R c are C2 alkyl groups. In some embodiments, R b and R c are C4 alkyl groups. In some embodiments, R b and R c are C6 alkyl groups. In some embodiments, R b and R c are C8 alkyl groups. In some embodiments, R 2 is -CR b Rc and R b and R c is C 10 alkyl. In some embodiments, R 2 is -CR b R c and R b and R c is C 12 alkyl. In some embodiments, R 2 is -CR b R c and R b is C 12 alkyl and R c is C 10 alkyl. In some embodiments, R 2 is -CR b R c and R b is C 10 alkyl and R c is C 12 alkyl. In some embodiments, R 2 is -CR b R c and R b is C8 alkyl and R c is C 10 alkyl. In some embodiments, R 2 is -CR b R c and R b is C6 alkyl and R c is C 10 alkyl. In some embodiments, R 2 is -CR b R c and R b is C4 alkyl and R c is C 10 alkyl. In some embodiments, R 2 is -CR b R c and R b is C2 alkyl and R c is C 10 alkyl. In some embodiments, R 2 is -CR b R c and R b is C1 alkyl and R c is C 10 alkyl. In some embodiments, R 2 is -CR b R c and Rb is C 10 alkyl and R c is C8 alkyl. In some embodiments, R 2 is -CR b R c and R b is C 10 alkyl and R c is C6 alkyl. In some embodiments, R 2 is -CR b R c and R b is C 10 alkyl and R c is C4 alkyl. In some embodiments, R 2 is -CR b R c and R b is C 10 alkyl and R c is C2 alkyl. In some embodiments, R 2 is -CR b R c and R b is C 10 alkyl and R c is C1 alkyl. In some embodiments, R 2 is -CR b R c and R b is C8 alkyl and R c is C8 alkyl. In some embodiments, R 2 is -CR b R c and R b is C6 alkyl and R c is C8 alkyl. In some embodiments, R 2 is -CR b R c and R b is C4 alkyl and R c is C8 alkyl. In some embodiments, R 2 is -CR b R c and R b is C2 alkyl and R c is C8 alkyl. In some embodiments, R 2 is -CR b R c and R b is C1 alkyl and R c is C8 alkyl. In some embodiments, R 2 is -CR bR c and R b is a C8 alkyl group and R c is a C6 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkyl group and R c is a C4 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkyl group and R c is a C2 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkyl group and R c is a C1 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkyl group and R c is a C6 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkyl group and R c is a C6 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkyl group and R c is a C6 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkyl group and R c is a C6 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C1 alkyl group and R c is a C6 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkyl group and R c is a C8 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkyl group and Rc is a C4 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkyl group and R c is a C2 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkyl group and R c is a C1 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkyl group and R c is a C4 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkyl group and R c is a C4 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkyl group and R c is a C4 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkyl group and R c is a C4 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C1 alkyl group and R c is a C4 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkyl group and R c is a C8 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkyl group and R c is a C6 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkyl group and R c is a C4 alkyl group. In some embodiments, R 2-CR b R c and R b is a C4 alkyl group and R c is a C2 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkyl group and R c is a C1 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkyl group and R c is a C2 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkyl group and R c is a C2 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkyl group and R c is a C2 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkyl group and R c is a C2 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C1 alkyl group and R c is a C2 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkyl group and R c is a C8 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkyl group and R c is a C6 alkyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkyl group and R c is a C4 alkyl group. In some embodiments, R 2 is -CR b R c and Rb is a C2 alkyl and R c is a C1 alkyl. In some embodiments, R 2 is -CR b R c and R b is a C8 alkyl and R c is a C1 alkyl. In some embodiments, R 2 is -CR b R c and R b is a C6 alkyl and R c is a C1 alkyl. In some embodiments, R 2 is -CR b R c and R b is a C4 alkyl and R c is a C1 alkyl. In some embodiments, R 2 is -CR b R c and R b is a C2 alkyl and R c is a C1 alkyl. In some embodiments, R 2 is -CR b R c and R b is a C1 alkyl and R c is a C1 alkyl. In some embodiments, R 2 is -CR b R c and R b is a C1 alkyl and R c is a C8 alkyl. In some embodiments, R 2 is -CR b R c and R b is a C1 alkyl and R c is a C6 alkyl. In some embodiments, R 2 is -CR b R c and R b is a C1 alkyl and R c is a C4 alkyl. In some embodiments, R 2 is -CR b R c and R b is a C1 alkyl and R c is a C2 alkyl.

[0290] In some embodiments, R 2 is -CR b R c 。In some embodiments, Rb and R c is a C2 alkenyl group. In some embodiments, R b and R c is a C4 alkenyl group. In some embodiments, R b and R c is a C6 alkenyl group. In some embodiments, R b and R c is a C8 alkenyl group. In some embodiments, R 2 is -CR b R c and R b and R c is C 10 alkenyl. In some embodiments, R 2 is -CR b R c and R b and R c is C 12 alkenyl. In some embodiments, R 2 is -CR b R c and R b is C 12 alkenyl and R c is C 10 alkenyl. In some embodiments, R 2 is -CR b R c and R b is C 10 alkenyl and R c is C 12 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C8 alkenyl group and R c is C 10 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C6 alkenyl group and R c is C 10 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C4 alkenyl group and R c is C 10 alkenyl. In some embodiments, R 2 is -CR b R c and R bis a C2 alkenyl group and R c is C 10 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C1 alkenyl group and R c is C 10 alkenyl. In some embodiments, R 2 is -CR b R c and R b is C 10 and R c is a C8 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is C 10 alkenyl and R c is a C6 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is C 10 alkenyl and R c is a C4 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is C 10 alkenyl and R c is a C2 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is C 10 alkenyl and R c is a C1 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkenyl group and R c is a C8 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkenyl group and R c is a C8 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkenyl group and R c is a C8 alkenyl group. In some embodiments, R 2 is -CRb R c and R b is a C2 alkenyl group and R c is a C8 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C1 alkenyl group and R c is a C8 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkenyl group and R c is a C6 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkenyl group and R c is a C4 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkenyl group and R c is a C2 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkenyl group and R c is a C1 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkenyl group and R c is a C6 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkenyl group and R c is a C6 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkenyl group and R c is a C6 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkenyl group and R c is a C6 alkenyl group. In some embodiments, R 2 is -CR b R c and R bis a C1 alkenyl and R c is a C6 alkenyl. In some embodiments, R 2 is -CR b R c and R b is C6 and R c is a C8 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C6 alkenyl and R c is a C4 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C6 alkenyl and R c is a C2 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C6 alkenyl and R c is a C1 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C8 alkenyl and R c is a C4 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C6 alkenyl and R c is a C4 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C4 alkenyl and R c is a C4 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C2 alkenyl and R c is a C4 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C1 alkenyl and R c is a C4 alkenyl. In some embodiments, R 2 is -CR b R c and R b is a C4 alkenyl and R c is a C8 alkenyl. In some embodiments, R2 is -CR b R c and R b is a C4 alkenyl group and R c is a C6 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkenyl group and R c is a C4 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkenyl group and R c is a C2 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkenyl group and R c is a C1 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkenyl group and R c is a C2 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkenyl group and R c is a C2 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkenyl group and R c is a C2 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkenyl group and R c is a C2 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C1 alkenyl group and R c is a C2 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkenyl group and R c is a C8 alkenyl group. In some embodiments, R 2 is -CR b R cAnd R b is a C2 alkenyl group and R c is a C6 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkenyl group and R c is a C4 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkenyl group and R c is a C1 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C8 alkenyl group and R c is a C1 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C6 alkenyl group and R c is a C1 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C4 alkenyl group and R c is a C1 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C2 alkenyl group and R c is a C1 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C1 alkenyl group and R c is a C1 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C1 alkenyl group and R c is a C8 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C1 alkenyl group and R c is a C6 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C1 alkenyl group and R cis a C4 alkenyl group. In some embodiments, R 2 is -CR b R c and R b is a C1 alkenyl group and R c is a C2 alkenyl group.

[0291] In some embodiments, R 2 is -C6H 13 、-C7H 15 、-C8H 17 、-C 10 H 21 、-C 11 H 23 、-C 12 H 25 、-C 13 H 27 、-C 14 H 29 、-C 15 H 31 、-C 16 H 33 、-C 17 H 35 、-C 18 H 36 、-C 19 H 39 、-C 20 H 41 、-C 21 H 43 、-C 22 H 45 、-C 23 H 47 、-C 24 H 49 、-C6H 12 、-C7H 14 、-C8H 16 、-C 10 H 20 、-C 11 H 21 、-C 12 H 24 、-C 13 H 26 、-C 14 H 28 、-C 15 H 30 、-C 16 H 32 、-C 17 H 34 、-C 18 H 35 、-C 19 H38 , -C 20 H 40 , -C 21 H 42 , -C 22 H 44 , -C 23 H 46 or -C 24 H 48 .

[0292] In some embodiments, R 6 is In some embodiments, p and q are 0. In some embodiments, p and q are 1. In some embodiments, p and q are 2. In some embodiments, p and q are 3. In some embodiments, p and q are 4. In some embodiments, p is 0 and q is 1. In some embodiments, p is 0 and q is 2. In some embodiments, p is 0 and q is 3. In some embodiments, p is 0 and q is 4. In some embodiments, p is 1 and q is 2. In some embodiments, p is 1 and q is 3. In some embodiments, p is 1 and q is 4. In some embodiments, p is 2 and q is 1. In some embodiments, p is 2 and q is 3. In some embodiments, p is 2 and q is 4. In some embodiments, p is 3 and q is 1. In some embodiments, p is 3 and q is 2. In some embodiments, p is 3 and q is 4. In some embodiments, p is 4 and q is 1. In some embodiments, p is 4 and q is 2. In some embodiments, p is 4 and q is 3.

[0293] In some embodiments, R 6 is In some embodiments, R 3 is In some embodiments, p, r, and q are 0. In some embodiments, p, r, and q are 1. In some embodiments, p, r, and q are 2. In some embodiments, p, r, and q are 3. In some embodiments, p, r, and q are 4. In some embodiments, p is 0, r is 0 and q is 1. In some embodiments, p is 0, r is 0 and q is 2. In some embodiments, p is 0, r is 0 and q is 3. In some embodiments, p is 0, r is 0 and q is 4. In some embodiments, p is 1, r is 0 and q is 2. In some embodiments, p is 1, r is 0 and q is 3. In some embodiments, p is 1, r is 0 and q is 4. In some embodiments, p is 2, r is 0 and q is 1. In some embodiments, p is 2, r is 0 and q is 3. In some embodiments, p is 2, r is 0 and q is 4. In some embodiments, p is 3, r is 0 and q is 1. In some embodiments, p is 3, r is 0 and q is 2. In some embodiments, p is 3, r is 0 and q is 4. In some embodiments, p is 4, r is 0 and q is 1. In some embodiments, p is 4, r is 0 and q is 2. In some embodiments, p is 4, r is 0 and q is 3. In some embodiments, p is 0, r is 1 and q is 1. In some embodiments, p is 0, r is 1 and q is 2. In some embodiments, p is 0, r is 1 and q is 3. In some embodiments, p is 0, r is 1 and q is 4. In some embodiments, p is 1, r is 1 and q is 2. In some embodiments, p is 1, r is 1 and q is 3. In some embodiments, p is 1, r is 1 and q is 4. In some embodiments, p is 2, r is 1 and q is 1. In some embodiments, p is 2, r is 1 and q is 3. In some embodiments, p is 2, r is 1 and q is 4. In some embodiments, p is 3, r is 1 and q is 1. In some embodiments, p is 3, r is 1 and q is 2. In some embodiments, p is 3, r is 1 and q is 4. In some embodiments, p is 4, r is 1 and q is 1. In some embodiments, p is 4, r is 1 and q is 2. In some embodiments, p is 4, r is 1 and q is 3.

[0294] In some embodiments, p is 0, r is 2 and q is 1. In some embodiments, p is 0, r is 2 and q is 2. In some embodiments, p is 0, r is 2 and q is 3. In some embodiments, p is 0, r is 2 and q is 4. In some embodiments, p is 1, r is 2 and q is 2. In some embodiments, p is 1, r is 2 and q is 3. In some embodiments, p is 1, r is 2 and q is 4. In some embodiments, p is 2, r is 2 and q is 1. In some embodiments, p is 2, r is 2 and q is 3. In some embodiments, p is 2, r is 2 and q is 4. In some embodiments, p is 3, r is 2 and q is 1. In some embodiments, p is 3, r is 2 and q is 2. In some embodiments, p is 3, r is 2 and q is 4. In some embodiments, p is 4, r is 2 and q is 1. In some embodiments, p is 4, r is 2 and q is 2. In some embodiments, p is 4, r is 2 and q is 3.

[0295] In some embodiments, p is 0, r is 3 and q is 1. In some embodiments, p is 0, r is 3 and q is 2. In some embodiments, p is 0, r is 3 and q is 3. In some embodiments, p is 0, r is 3 and q is 4. In some embodiments, p is 1, r is 3 and q is 2. In some embodiments, p is 1, r is 3 and q is 3. In some embodiments, p is 1, r is 3 and q is 4. In some embodiments, p is 2, r is 3 and q is 1. In some embodiments, p is 2, r is 3 and q is 3. In some embodiments, p is 2, r is 3 and q is 4. In some embodiments, p is 3, r is 3 and q is 1. In some embodiments, p is 3, r is 3 and q is 2. In some embodiments, p is 3, r is 3 and q is 4. In some embodiments, p is 4, r is 3 and q is 1. In some embodiments, p is 4, r is 3 and q is 2. In some embodiments, p is 4, r is 3 and q is 3.

[0296] In some embodiments, p is 0, r is 4 and q is 1. In some embodiments, p is 0, r is 4 and q is 2. In some embodiments, p is 0, r is 4 and q is 3. In some embodiments, p is 0, r is 4 and q is 4. In some embodiments, p is 1, r is 4 and q is 2. In some embodiments, p is 1, r is 4 and q is 3. In some embodiments, p is 1, r is 4 and q is 4. In some embodiments, p is 2, r is 4 and q is 1. In some embodiments, p is 2, r is 4 and q is 3. In some embodiments, p is 2, r is 4 and q is 4. In some embodiments, p is 3, r is 4 and q is 1. In some embodiments, p is 3, r is 4 and q is 2. In some embodiments, p is 3, r is 4 and q is 4. In some embodiments, p is 4, r is 4 and q is 1. In some embodiments, p is 4, r is 4 and q is 2. In some embodiments, p is 4, r is 4 and q is 3.

[0297] In some embodiments, M is N.

[0298] In some embodiments, M is CH.

[0299] In some embodiments, M is

[0300] In some embodiments, M is

[0301] In some embodiments, M is

[0302] In some embodiments, M is

[0303] In some embodiments, M is

[0304] In some embodiments, M is

[0305] In some embodiments, D is N.

[0306] In some embodiments, D is In some embodiments, the compounds of formulas Da, Db, Dc, Y, B, C, I - IX may be selected from:

[0307]

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314] or a pharmaceutically acceptable salt thereof.

[0315] Compositions are described herein that comprise a compound of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof; and an active agent.

[0316] Composition

[0317] The compositions described herein comprise a compound of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof, and an active agent. The compositions may further comprise an excipient useful for a variety of medical and non-medical applications. For example, the pharmaceutical compositions described herein can be used to deliver an effective amount of an agent to a subject in need thereof. The compositions described herein can be used for non-medical applications such as, for example, as emulsions or emulsifying agents and can be used, for example, as food ingredients, for fire extinguishing, for disinfecting surfaces, for oil removal, etc.

[0318] In some embodiments, the composition can be a lipid nanoparticle dispersion, a liposome formulation, a lipid emulsion, or any combination thereof, comprising a compound of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof.

[0319] In one aspect, the present disclosure provides a composition comprising: a lipid nanoparticle comprising a compound of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof; and an active agent.

[0320] In certain embodiments, the composition can comprise an agent described herein. For example, in certain embodiments, the agent is any chemical compound to be administered to a subject that can be delivered using a particle or nanoparticle described herein. The agent can be an organic molecule (e.g., a therapeutic agent, a drug), an inorganic molecule, a small molecule, an organometallic compound, a metal, a nucleic acid, a protein, an amino acid, a peptide, a polypeptide, a polynucleotide, a targeting agent, an isotopically labeled organic or inorganic molecule, a vaccine, an immunological agent, or an agent for bioprocessing. In certain embodiments, the agent is a polynucleotide. In some embodiments, the agent comprises an mRNA encoding at least one antigenic polypeptide or an immunogenic fragment thereof capable of inducing an immune response against the antigenic polypeptide.

[0321] In some embodiments, the mRNA encoding at least one antigenic polypeptide or an immunogenic fragment thereof capable of inducing an immune response against the antigenic polypeptide is encapsulated by a nanoparticle.

[0322] In some aspects, the present disclosure provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and nanoparticles, the nanoparticles comprising a compound of formula Da, Db, Db, Y, B, C, I-IX or any combination thereof; and an mRNA encoding at least one antigenic polypeptide or an immunogenic fragment thereof capable of inducing an immune response against the antigenic polypeptide.

[0323] nanoparticles

[0324] The present disclosure describes lipid nanoparticles comprising greater than 0% to 50% of the compounds described herein (e.g., a compound of formula Da, Db, Db, Y, B, C, I-IX or any combination thereof); 0% to 50% cationic lipid, 0% to 80% ionizable lipid; greater than 0% to 5% polyethylene glycol-lipid; greater than 0% to 40% helper lipid; 0% to 80% sterol; and an active agent encapsulated in the nanoparticles.

[0325] The present disclosure also describes a composition comprising an effective amount of the lipid nanoparticles described herein and a pharmaceutically acceptable carrier.

[0326] In one aspect, the present disclosure provides a nanoparticle comprising: a compound of formula Da, Db, Db, Y, B, C, I-IX or any combination thereof; and optionally a cationic lipid, an ionizable lipid, a helper lipid; a polyethylene glycol-lipid; a sterol or any combination thereof; and an active agent encapsulated in the nanoparticles.

[0327] In one aspect, the present disclosure provides a nanoparticle comprising: a compound of formula Da, Db, Db, Y, B, C, I-IX or any combination thereof; and a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticles.

[0328] In one aspect, the present disclosure provides a nanoparticle comprising: a compound of formula Da, Db, Db, Y, B, C, I-IX or any combination thereof; a helper lipid; a polyethylene glycol-lipid; and a sterol; and an active agent encapsulated in the nanoparticles.

[0329] In one aspect, the present disclosure provides a nanoparticle comprising: a compound of formula Da, Db, Db, Y, B, C, I-IX or any combination thereof; a cationic lipid; a helper lipid; a polyethylene glycol-lipid; and a sterol; and an active agent encapsulated in the nanoparticles.

[0330] On the one hand, the present disclosure provides a nanoparticle comprising: compounds of Da, Db, Db, Y, B, C, I-IX or any combination thereof; ionizable lipids, helper lipids; polyethylene glycol-lipids; and sterols; and an active agent encapsulated in the nanoparticle.

[0331] On the one hand, the present disclosure provides a nanoparticle comprising: compounds of Da, Db, Db, Y, B, C, I-IX or any combination thereof; ionizable lipids, and cationic lipids; and an active agent encapsulated in the nanoparticle.

[0332] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula Da; and helper lipids; polyethylene glycol-lipids; and / or sterols; and an active agent encapsulated in the nanoparticle.

[0333] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula Db; and helper lipids; polyethylene glycol-lipids; and / or sterols; and an active agent encapsulated in the nanoparticle.

[0334] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula Dc; and helper lipids; polyethylene glycol-lipids; and / or sterols; and an active agent encapsulated in the nanoparticle.

[0335] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula Y; and helper lipids; polyethylene glycol-lipids; and / or sterols; and an active agent encapsulated in the nanoparticle.

[0336] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula A; and helper lipids; polyethylene glycol-lipids; and / or sterols; and an active agent encapsulated in the nanoparticle.

[0337] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula B; and helper lipids; polyethylene glycol-lipids; and / or sterols; and an active agent encapsulated in the nanoparticle.

[0338] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula C; and helper lipids; polyethylene glycol-lipids; and / or sterols; and an active agent encapsulated in the nanoparticle.

[0339] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula I; and helper lipids; polyethylene glycol-lipids; and / or sterols; and an active agent encapsulated in the nanoparticle.

[0340] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula II; and a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0341] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula III; and a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0342] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula IV; and a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0343] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula V; and a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0344] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula VI; and a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0345] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula VII; and a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0346] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula VIII; and a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0347] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula IX; and a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0348] On the one hand, the present disclosure provides a nanoparticle comprising: Da, Db, Db, Y, B, C, compounds of I-IX or any combination thereof; and optionally an ionizable lipid, a helper lipid; a polyethylene glycol-lipid; a sterol or any combination thereof; and an active agent encapsulated in the nanoparticle.

[0349] On the one hand, the present disclosure provides a nanoparticle comprising: Da, Db, Db, Y, B, C, compounds of I-IX or any combination thereof; and an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0350] On the one hand, the present disclosure provides a nanoparticle comprising: Da, Db, Db, Y, B, C, I-IX compounds or any combination thereof; an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and a sterol; and an active agent encapsulated in the nanoparticle.

[0351] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula Da; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0352] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula Db; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0353] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula Dc; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0354] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula Y; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0355] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula B; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0356] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula C; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0357] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula I; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0358] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of formula II; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0359] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula III; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0360] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula IV; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0361] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula V; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0362] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula VI; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0363] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula VII; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0364] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula VIII; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0365] On the one hand, the present disclosure provides a nanoparticle comprising: a compound of Formula IX; and a cationic lipid, an ionizable lipid; a helper lipid; a polyethylene glycol-lipid; and / or a sterol; and an active agent encapsulated in the nanoparticle.

[0366] Various compounds of Formulae Da, Db, Db, Y, B, C or I-IX are described in the above-mentioned compound section. In some embodiments, the nanoparticle comprises a compound of Formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof in a molar ratio greater than 0% to 50%; and an active agent encapsulated in the nanoparticle.

[0367] In some embodiments, the compounds of formulae Da, Db, Db, Y, B, C or I-IX may be present in a molar ratio of at least greater than 0% (e.g., at least 5%, at least 10%, at least 20%, at least 30%, at least 40% or at least 50%). In some embodiments, the compounds of formulae Da, Db, Db, Y, B, C or I-IX may be present in a molar ratio of 50% or less (e.g., 50% or less, 40% or less, 30% or less, 20% or less, 10% or less, 5% or less, 1% or less or 0.5% or less).

[0368] The compounds of formulae Da, Db, Db, Y, B, C or I-IX may be present in a molar ratio ranging from any of the above minima to any of the above maxima. For example, in some embodiments, the compounds of formulae Da, Db, Db, Y, B, C or I-IX may be present in a molar ratio of greater than 0% to 50% (e.g., greater than 0% to 40%, greater than 0% to 30%, greater than 0% to 20%, greater than 0% to 10%, greater than 0% to 5%, greater than 0% to 1%, greater than 0% to 0.5%, 1% to 40%, 1% to 50%, 1% to 30%, 1% to 20%, 1% to 10%, 1% to 5%, 5% to 50%, 5% to 40%, 5% to 30%, 5% to 20%, 5% to 10%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 20%, 20% to 50%, 20% to 40%, 20% to 30%, 20% to 40%, 30% to 50%, 30% to 40% or 40% to 50%).

[0369] Cationic lipid

[0370] Suitable cationic lipids can include, but are not limited to, 2-amino-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-2-{[(9Z,2Z)-octadeca-9,12-dien-1-yloxy]methyl}propan-1-ol (Compound 1 in US20130150625); 2-amino-3-[(9Z)-octadec-9-en-1-yloxy]-2-{[(9Z)-octadec-9-en-1-yloxy]methyl}propan-1-ol (Compound 2 in US20130150625); 2-amino-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-2-[(octyloxy)methyl]propan-1-ol (Compound 3 in US20130150625); and 2-(dimethylamino)-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-2-{[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]methyl}propan-1-ol (Compound 4 in US20130150625); or any pharmaceutically acceptable salt or stereoisomer thereof. As a non-limiting example, the cationic lipid can be selected from (20Z,23Z)—N,N-dimethylnonacosa-20,23-dien-10-amine, (17Z,20Z)—N,N-dimethylhexacos-17,20-dien-9-amine, (1Z,19Z)—N5N-dimethylpentacos-16,19-dien-8-amine, (13Z,16Z)—N,N-dimethyl-docos-13,16-dien-5-amine, (12Z,15Z)—N,N-dimethylheneicos-12,15-dien-4-amine, (14Z,17Z)—N,N-dimethyltricos-14,17-dien-6-amine, (15Z,18Z)—N,N-dimethyltetracos-15,18-dien-7-amine, (18Z,21Z)—N,N-dimethylheptacos-18,21-dien-10-amine, (15Z,18Z)—N,N-dimethyltetracos-15,18-dien-5-amine, (14Z,17Z)—N,N-dimethyltricos-14,17-dien-4-amine, (19Z,22Z)—N,N-dimethyloctacos-19,22-dien-9-amine, (18Z,21Z)—N,N-dimethylheptacos-18,21-dien-8amine, (17Z,20Z)—N,N-dimethylhexacos-17,20-dien-7-amine, (16Z,19Z)—N,N-dimethylpentacos-16,19-dien-6-amine, (22Z,25Z)—N,N-dimethylhentriaconta-22,25-dien-10-amine, (21Z,24Z)—N,N-dimethyltriaconta-21,24-dien-9-amine, (18Z)—N,N-dimethylheptacos-18-en-10-amine, (17Z)-N,N-dimethylhexacos-17-en-9-amine, (19Z,22Z)-N,N-dimethyloctacos-19,22-dien-7-amine, N,N-dimethylheptacosan-10-amine, (20Z,23Z)-N-ethyl-N-methylnonacos-20,23-dien-10-amine, 1-[(11Z,14Z)-1-nonylicos-11,14-dien-1-yl]pyrrolidine, (20Z)-N,N-dimethylheptacos-20-en-10-amine, (15Z)-N,N-dimethylheptacos-15-en-10-amine, (14Z)-N,N-dimethylnonacos-14-en-10-amine, (17Z)-N,N-dimethylnonacos-17-en-10-amine, (24Z)-N,N-dimethyltritriacont-24-en-10-amine, (20Z)-N,N-dimethylnonacos-20-en-10-amine, (22Z)-N,N-dimethylhentriacont-22-en-10-amine, (16Z)-N,N-dimethylpentacos-16-en-8-amine, (12Z,15Z)-N,N-dimethyl-2-nonylheneicos-12,15-dien-1-amine, (13Z,16Z)-N,N-dimethyl-3-nonyl-docos-13,16-dien-1-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]heptadecane-8-amine, 1-[(1S,2R)-2-hexylcyclopropyl]-N,N-dimethylnonadecane-10-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]nonadecane-10-amine, N,N-dimethyl-21-[(1S,2R)-2-octylcyclopropyl]heneicosane-10-amine, N,N-dimethyl-1-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]methyl}cyclopropyl]nonadecane-10-amine, N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]hexadecane-8-amine, N,N-dimethyl-[(1R,2S)-2-undecylcyclopropyl]tetradecane-5-amine, N,N-dimethyl-3-{7-[(1S,2R)-2-octylcyclopropyl]heptyl}dodecane-1-amine, 1-[(1R,2S)-2-heptylcyclopropyl]-N,N-dimethyloctadecane-9-amine, 1-[(1S,2R)-2-decylcyclopropyl]-N,N-dimethylpentadecane-6-amine, N,N-dimethyl-1-R1S,2R)-2-octylcyclopropylpentadecane-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-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine, N,N-dimethyl-1-(nonyloxy)-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)-octadeca-6,9,12-trien-1-yloxy]-3-(octyloxy)propan-2-amine, (2S)-1-[(11Z,14Z)-eicosa-11,14-dien-1-yloxy]-N,N-dimethyl-3-(pentyloxy)propan-2-amine, (2S)-1-(hexyloxy)-3-[(11Z,14Z)-eicosa-11,14-dien-1-yloxy]-N,N-dimethylpropan-2-amine, 1-[(11Z,14Z)-eicosa-11,14-dien-1-yloxy]-N,N-dimethyl-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-dimethylpropan-2-amine, (2S)-1-[(13Z)-docos-13-en-1-yloxy]-3-(hexyloxy)-N,N-dimethylpropan-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-dimethyl-1-[(1-methyloctyl)oxy]-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine, (2R)-1-[(3,7-dimethyloctyl)oxy]-N,N-dimethyl-3-((9Z,12Z)-octadeca-9,12-dien-1-yloxy)propan-2-amine, N,N-dimethyl-1-(octyloxy)-3-({8-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]methyl}cyclopropyl]octyl}oxy)propan-2-amine, N,N-dimethyl-1-({8-(2-octylcyclopropyl)octyl}oxy)-3-(octyloxy)propan-2-amine, and (11E,20Z,23Z)-N,N-dimethylnonacosa-11,20,23-triene-10-amine or a pharmaceutically acceptable salt or stereoisomer thereof. In some embodiments, the lipid can be a cationic lipid such as, but not limited to, DLin-DMA, DLin-D-DMA, DLin-MC3-DMA, DLin-KC2-DMA, DODMA, and aminoalcohol lipids. The aminoalcohol cationic lipid can be a lipid described in U.S. Patent Publication No. US20130150625, which is incorporated herein by reference in its entirety, and / or a lipid made by the methods described in said U.S. Patent Publication. In some embodiments, the nanoparticles described herein can include amine cationic lipids such as those described in International Patent Application No. WO2013059496, the content of which is incorporated herein by reference in its entirety. In some embodiments, the cationic lipid can have an amino-amine or amino-amide moiety.,

[0371] In some embodiments, the cationic lipid can be a low molecular weight cationic lipid such as those described in U.S. Patent Application No. 2013 / 0090372, the content of which is incorporated herein by reference in its entirety.

[0372] Ionizable lipid

[0373] Exemplary ionizable lipids are described in U.S. Patent Publication Nos. U.S. 2016 / 0311759, U.S. 2015 / 0376115, U.S. 2016 / 0151284, U.S. 2017 / 0210697, U.S. 2015 / 0140070, U.S. 2013 / 0178541, U.S. 2013 / 0303587, U.S. 2015 / 0141678, U.S. 2015 / 0239926, U.S. 2016 / 0376224, U.S. 2017 / 0119904, U.S. 2012 / 0149894, U.S. 2015 / 0057373, U.S. 2013 / 0090372, U.S. 2013 / 0274523, U.S. 2013 / 0274504, U.S. 2013 / 0274504, U.S. 2009 / 0023673, U.S. 2012 / 0128760, U.S. 2010 / 03241240, U.S. 2014 / 0200257, U.S. 2015 / 0203446, U.S. 2018 / 0005363, U.S. 2014 / 0308304, U.S. 2013 / 0338210, U.S. 2012 / 0101148, U.S. 2012 / 0027796, U.S. 2012 / 0058144, U.S. 2013 / 0323269, U.S. 2011 / 0117125, U.S. 2011 / 0256175, U.S. 2012 / 0202871, U.S. 2011 / 0076335, U.S. 2006 / 0083780, U.S. 2013 / 0123338, U.S. 2015 / 0064242, U.S. 2006 / 0051405, U.S. 2013 / 0065939, U.S. 2006 / 0008910, U.S. 2003 / 0022649, U.S. 2010 / 0130588, U.S. 2013 / 0116307, U.S. 2010 / 0062967, U.S. 2013 / 0202684, U.S. 2014 / 0141070, U.S. 2014 / 0255472, U.S. 2014 / 0039032, U.S. 2018 / 0028664, U.S. 2016 / 0317458, U.S. 2013 / 0195920, U.S. 2022 / 0062175, U.S.No. 2021 / 0121411, No. U.S. 2022 / 0009878, No. U.S. 2022 / 0040325, No. U.S. 2012 / 61657480, No. U.S. 2016 / 0074514, No. U.S. 2013 / 0330401, No. U.S. 2019 / 0185410, No. U.S. 2012 / 61617468, No. U.S. 2019 / 0032087, No. U.S. 2015 / 62184188, No. U.S. 2019 / 0127318, No. U.S. 2021 / 0002813, No. U.S. 2020 / 0345641, No. U.S. 2014 / 61944336, No. U.S. 2012 / 61657480, No. U.S. 2021 / 0059953, No. U.S. 2022 / 0162521, No. U.S. 2022 / 0235377, No. U.S. 2018 / 0085474, No. U.S. 2018 / 0000953, No. U.S. 2020 / 0129445, No. U.S. 2021 / 0145982, No. U.S. 2021 / 0378980, No. U.S. 2020 / 0254086, No. U.S. 2021 / 0346306, and No. U.S. 2018 / 0000953, the entire disclosures of all of which are incorporated herein by reference.

[0374] In some embodiments, the nanoparticles comprise an ionizable lipid in a molar ratio of 0% to 80%. In some embodiments, the ionizable lipid can be present in a molar ratio of at least 0% (e.g., at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80%). In some embodiments, the ionizable lipid can be present in a molar ratio of 80% or less (e.g., 80% or less, 70% or less, 60% or less, 50% or less, 40% or less, 30% or less, 20% or less, 10% or less, 5% or less, 1% or less, or 0.5% or less).

[0375] The ionizable lipid can be present in a molar ratio ranging from any of the above minima to any of the above maxima. For example, in some embodiments, the ionizable lipid can be present in a molar ratio of 0% to 80% (e.g., greater than 0% to 80%, greater than 0% to 70%, greater than 0% to 60%, greater than 0% to 50%, greater than 0% to 40%, greater than 0% to 30%, greater than 0% to 20%, greater than 0% to 10%, greater than 0% to 5%, greater than 0% to 1%, greater than 0% to 0.5%, 1% to 30%, 1% to 20%, 1% to 10%, 1% to 5%, 5% to 30%, 5% to 20%, 5% to 10%, 10% to 30%, 10% to 20%, 20% to 30%, 20% to 40%, or 30% to 40%).

[0376] Helper lipid

[0377] In some embodiments, the nanoparticles comprise helper lipids. In some embodiments, the helper lipids can be non-cationic lipids. In some embodiments, the non-cationic lipids can include, but are not limited to, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1-stearoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (SOPE), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dioleoyl-5 / 7-glycero-3-phosphate-(1'-rac-glycerol) (DOPG), 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine (PLPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-stearoyl-2-myristoyl-sn-glycero-3-phosphocholine (SMPC), or combinations thereof. In one embodiment, the non-cationic lipid is 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE). In one embodiment, the non-cationic lipid is 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE). In one embodiment, the non-cationic lipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). In one embodiment, the non-cationic lipid is 1-stearoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (SOPE). Although several non-cationic lipids are described herein, additional non-cationic lipids can be used in combination with the compounds disclosed herein.

[0378] In some embodiments, the helper lipids can be present in a molar ratio of at least 0% (e.g., at least 5%, at least 10%, at least 20%, at least 30%, or at least 40%). In some embodiments, the helper lipids can be present in a molar ratio of 40% or less (e.g., 30% or less, 20% or less, 10% or less, 5% or less, 1% or less, or 0.5% or less).

[0379] The co-lipid can be present in a molar ratio ranging from any of the above minima to any of the above maxima. For example, in some embodiments, the co-lipid can be present in a molar ratio of 0% to 40% (e.g., greater than 0% to 30%, greater than 0% to 20%, greater than 0% to 10%, greater than 0% to 5%, greater than 0% to 1%, greater than 0% to 0.5%, 1% to 30%, 1% to 20%, 1% to 10%, 1% to 5%, 5% to 30%, 5% to 20%, 5% to 10%, 10% to 30%, 10% to 20%, 20% to 30%, 20% to 40%, or 30% to 40%).

[0380] Polyethylene glycol lipid

[0381] In some embodiments, the nanoparticles include polyethylene glycol-lipid (PEG-lipid). The PEG-lipid is incorporated to form a hydrophilic outer layer and stabilize the particles. Non-limiting examples of PEG-lipids include PEG-modified lipids such as PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol. Representative polyethylene glycol-lipids include DMG-PEG, DLPE-PEG, DMPE-PEG, DPPC-PEG, and DSPE-PEG. In one embodiment, the polyethylene glycol-lipid is 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG). In one embodiment, the polyethylene glycol-lipid is 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol-2000 (DMG-PEG2000). DMG-PEGXXXX means 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol-XXXX, where XXXX represents the molecular weight of the polyethylene glycol moiety, e.g., DMG-PEG2000 or DMG-PEG5000.

[0382] In some embodiments, the polyethylene glycol-lipid can be present in a molar ratio of at least 0% (e.g., at least 0.25%, at least 0.5%, at least 0.75%, at least 1%, at least 1.5%, at least 2%, at least 3%, at least 4%, or at least 5%). In some embodiments, the polyethylene glycol-lipid can be present in a molar ratio of 5% or less (e.g., 4% or less, 3% or less, 2% or less, 1% or less, or 0.5% or less).

[0383] The polyethylene glycol lipid can be present in a molar ratio ranging from any of the above minima to any of the above maxima. For example, in some embodiments, the polyethylene glycol lipid can be present in a molar ratio of 0% to 5% (e.g., greater than 0% to 4%, greater than 0% to 3%, greater than 0% to 2%, greater than 0% to 1%, greater than 0% to 0.5%, 1% to 5%, 1% to 4%, 1% to 3%, 1% to 2%, 2% to 5%, 2% to 4%, 2% to 3%, 3% to 5%, 3% to 4%, or 4% to 5%). In one embodiment, the molar ratio of polyethylene glycol-lipid is 0.75%.

[0384] Sterol

[0385] In some embodiments, the nanoparticles include sterols. Sterols are well known to those skilled in the art and generally refer to compounds having a cyclopentanoperhydrophenanthrene ring system and having one or more OH substituents. Examples of sterols include, but are not limited to, cholesterol, campesterol, ergosterol, sitosterol, etc.

[0386] In some embodiments, the sterol is selected from cholesteryl lipids. In some embodiments, one or more cholesteryl lipids are selected from cholesterol, polyethylene glycolylated cholesterol, DC-Choi (N,N-dimethyl-N-ethylcarboxamide cholesterol), 1,4-bis(3-N-oleylaminopropyl)piperazine, or combinations thereof.

[0387] Sterols can tune the permeability and fluidity of the particles based on their function in cell membranes. In one embodiment, the sterol is cholesterol.

[0388] In some embodiments, the molar ratio of sterol is at least 0% (e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, or at least 80%). In some embodiments, the molar ratio of sterol is 80% or less (e.g., 70% or less, 60% or less, 50% or less, 40% or less, 30% or less, 20% or less, 10% or less, 5% or less, or 1% or less).

[0389] The sterol can be present in a molar ratio ranging from any of the above - mentioned minimum values to any of the above - mentioned maximum values. In some embodiments, the sterol can be present in a molar ratio of 0% to 80% (e.g., greater than 0% to 70%, greater than 0% to 60%, greater than 0% to 50%, greater than 0% to 40%, greater than 0% to 30%, greater than 0% to 20%, greater than 0% to 10%, greater than 0% to 5%, greater than 0% to 1%, 5% to 70%, 5% to 60%, 5% to 50%, 5% to 40%, 5% to 30%, 5% to 20%, 5% to 10%, 10% to 70%, 10% to 60%, 10% to 50%, 10% to 40%, 10% to 30%, 10% to 20%, 20% to 70%, 20% to 60%, 20% to 50%, 20% to 40%, 20% to 30%, 30% to 70%, 30% to 60%, 30% to 50%, 30% to 40%, 40% to 70%, 40% to 60%, 40% to 50%, 50% to 70%, 50% to 60%, 60% to 70%, 60% to 80%, 70% to 80%, 10% to 80%, 20% to 80%, 30% to 80%, 40% to 80%, or 50% to 80%). In one embodiment, the nanoparticle comprises a sterol in a molar ratio of 40%.

[0390] In one embodiment, the nanoparticle can comprise 1% to 30% of the compounds described herein; 1% to 2% of a polyethylene glycol lipid; 10% to 50% of an ionizable lipid; 8% to 12% of a helper lipid; 35% to 40% of a sterol; and an active agent encapsulated in the nanoparticle.

[0391] In one embodiment, the present disclosure provides a nanoparticle comprising:

[0392] Compounds of formulae Da, Db, Db, Y, B, C, I - IX or any combination thereof.

[0393] In some embodiments, nanoparticles are described that comprise:

[0394] Compounds of formulae Da, Db, Db, Y, B, C, I - IX or any combination thereof;

[0395] 1,2 - dioleoyl - sn - glycero - 3 - phosphoethanolamine (DOPE);

[0396] 1,2 - dimyristoyl - sn - glycerol, methoxypolyethylene glycol (DMG - PEG 2000 ); and

[0397] Cholesterol.

[0398] In one embodiment, the present disclosure provides a nanoparticle comprising:

[0399] Compounds of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof;

[0400] 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE);

[0401] 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG 2000 ); and

[0402] Cholesterol.

[0403] In one embodiment, the present disclosure provides a nanoparticle comprising:

[0404] Compounds of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof;

[0405] 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC);

[0406] 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG 2000 ); and

[0407] Cholesterol.

[0408] In one embodiment, the present disclosure provides a nanoparticle comprising:

[0409] Compounds of formula YDa, Db, Db, Y, B, C, I-IX or any combination thereof;

[0410] 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE);

[0411] 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG 2000 ); and

[0412] Cholesterol.

[0413] In some embodiments, nanoparticles are described that comprise:

[0414] Compounds of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof;

[0415] Ionizable lipid;

[0416] 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE);

[0417] 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG 2000 ); and

[0418] cholesterol.

[0419] In one embodiment, the present disclosure provides a nanoparticle comprising:

[0420] a compound of formula Da, Db, Db, Y, B, C, I-IX or any combination thereof;

[0421] an ionizable lipid;

[0422] 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE);

[0423] 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG 2000 ); and

[0424] cholesterol.

[0425] In one embodiment, the present disclosure provides a nanoparticle comprising:

[0426] a compound of formula Da, Db, Db, Y, B, C, I-IX or any combination thereof;

[0427] an ionizable lipid;

[0428] 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC);

[0429] 1,2-dimyristoyl-sn-glycerol, methoxypolyethylene glycol (DMG-PEG 2000 ); and

[0430] cholesterol.

[0431] In one embodiment, the present disclosure provides a nanoparticle comprising:

[0432] a compound of formula Da, Db, Db, Y, B, C, I-IX or any combination thereof;

[0433] an ionizable lipid;

[0434] 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE);

[0435] 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG 2000 ); and

[0436] cholesterol.

[0437] In one embodiment, the nanoparticles may further comprise an active agent. In one embodiment, the nanoparticles may further comprise a therapeutic agent. In one embodiment, the nanoparticles may further comprise a diagnostic agent. In one embodiment, the nanoparticles may further comprise a prophylactic agent.

[0438] Active agent

[0439] As used herein, "active agent" refers to a therapeutic agent, a diagnostic agent, or a prophylactic agent. As discussed herein, the therapeutic agent may be released in a bioactive form from the disclosed compounds, compositions, and systems.

[0440] It should also be understood that, as used herein, the term "therapeutic agent" refers to one or more therapeutic agents, active ingredients or substances that can be used to treat medical conditions. Therapeutic agents include any synthetic or naturally occurring bioactive compounds or substance compositions that, when administered to an organism (human or non-human animal), induce desired pharmacological, immunogenic and / or physiological effects through local and / or systemic action. The term thus encompasses those compounds or chemicals that are traditionally regarded as including drugs, vaccines and biopharmaceuticals that include molecules such as proteins, peptides, hormones, nucleic acids, gene constructs, etc. Examples of therapeutic agents are described in well-known references such as the Merck Index (14th Edition), the Physicians' Desk Reference (64th Edition) and The Pharmacological Basis of Therapeutics (12th Edition), and the therapeutic agents include, but are not limited to, drugs; vitamins and minerals such as essential amino acids, calcium, iron, potassium, zinc, vitamin B12, etc.; substances for treating, preventing, diagnosing, curing or alleviating diseases or disorders; substances that affect the structure or function of the body, or prodrugs that become bioactive or more active when placed in a physiological environment. For example, the term "therapeutic agent" includes compounds or compositions for all major therapeutic areas, including but not limited to adjuvants; antimicrobial agents (including antibiotics, antiviral agents, antiparasitic agents and antifungal agents), anti-inflammatory agents (including steroids and non-steroidal anti-inflammatory agents), anticoagulants, ophthalmic medications, gastrointestinal medications, antiplatelet agents and preservatives, steroid agents, antitumor agents, anticancer agents, antigens, antibodies (e.g., cetuximab, anti-CD24 antibody, panitumumab and bevacizumab), contraceptives, progesterones, anticholinergic agents, nutritional agents, analgesics and analgesic combinations such as acetaminophen, acetylsalicylic acid, etc.; anesthetics such as lidocaine, xylocaine, etc.; anorectic drugs such as dextrothyroxine, phendimetrazine tartrate, etc.; antiepileptic drugs, local and general anesthesia, hypnotics, sedatives, antipsychotic agents, psychotropic agents, antidepressants such as isocarboxazid, amoxapine, etc.; antianxiety agents, antagonists, neuronal blockers, anticholinergic and cholinergic agents, antimuscarinic and muscarinic agents, antiparkinsonian drugs, anti-Alzheimer's agents, antiadrenergic agents, antiarrhythmic agents, antihypertensive agents, hormones such as insulin, progesterone, estrogen, corticosteroids, glucocorticoids, androgens, etc.; and nutritional supplements, gout medications such as methylprednisolone, ibuprofen, etc.; anti-asthma medications such as terbutaline sulfate, theophylline, ephedrine, etc.;Antispasmodics such as phenytoin sodium, diazepam, etc.; anti-allergy drugs, antihistamines such as diphenhydramine hydrochloride, chlorpheniramine maleate, etc.; anti-nausea drugs, anti-tumor drugs, antipruritics, antipyretics; antispasmodics such as belladonna alkaloids, dicyclomine hydrochloride, etc.; cardiovascular drugs such as prazosin hydrochloride, nitroglycerin, propranolol hydrochloride, hydralazine hydrochloride, pancreatic lipase, succinate dehydrogenase, etc.; vasoactive agents, cardiovascular preparations (including calcium channel blockers, β-blockers, β-agonists and antiarrhythmic drugs), antihypertensive drugs, diuretics such as furosemide, spironolactone, etc.; vasodilators; central nervous system stimulants; cough and cold preparations; decongestants; diagnostic agents; bone growth stimulants and bone resorption inhibitors; muscle relaxants; psychostimulants; sedatives; tranquilizers such as 72uteinizi, diazepam, chlorpromazine hydrochloride, reserpine, chlordiazepoxide hydrochloride, etc.; anti-peptic ulcer drugs such as ranitidine hydrochloride, cimetidine hydrochloride, etc.; anti-asthma drugs, antidiarrheals, weight loss agents, antithrombotic agents, antitussives, anti-uremia drugs, anti-angina drugs, appetite suppressants, expectorants, hyperglycemic drugs, hypoglycemic drugs, thyroid and anti-thyroid drugs, tissue growth agents, uterine relaxants, immunomodulators, including cytokines, interleukins, interferons, colony-stimulating factors, tumor necrosis factors, etc.; immunosuppressants such as rapamycin, tacrolimus, etc.; immunopharmaceuticals; antigens, factors, growth factors, amino acids, peptides and proteins and their fragments (whether naturally occurring, chemically synthesized or recombinantly produced) such as LHRH, somatostatin, calcitonin, growth hormone, glucagon-like peptide, growth releasing factor, angiotensin, FSH, EGF, bone morphogenetic protein (BMP), erythropoietin (EPO), interferon, interleukin, collagen, fibrinogen, insulin, factor VIII, factor IX; α-glucosidase, Vasopressin, ACTH, human serum albumin, γ-globulin, structural proteins, blood product proteins, complex proteins, antigens or antigen polypeptides, enzymes, antibodies, monoclonal antibodies, etc.; and nucleic acid molecules (polymeric forms of two or more nucleotides, polynucleotides, ribonucleotides (RNA) or deoxyribonucleotides (DNA), including double-stranded and single-stranded molecules, gene constructs, expression vectors, antisense molecules, etc.), small molecules (e.g., doxorubicin) and other bioactive macromolecules such as proteins and enzymes. The agent can be a bioactive agent for medical (including veterinary) applications and agriculture (such as plants) and other fields. In certain embodiments of the present disclosure, the agent to be delivered can be a mixture of active agents.

[0441] Representative examples of antibiotics include amikacin, amoxicillin, ampicillin, atovaquone, azithromycin, aztreonam, bacitracin, carbenicillin, cefadroxil, cefazolin, cefdinir, cefditoren, cefepime, cefiderocol, cefoperazone, cefotetan, cefoxitin, cefotaxime, cefpodoxime, cefprozil, ceftaroline, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, chloramphenicol, colistimethate, cefuroxime, cephalexin, cephradine, cilastatin, cinoxacin, ciprofloxacin, clarithromycin, clindamycin, dalbavancin, dalfopristin, daptomycin, demeclocycline, dicloxacillin, doripenem, doxycycline, eravacycline, ertapenem, erythromycin, fidaxomicin, 73uteinizin, gatifloxacin, 73uteinizing73, Gentamicin, Imipenem, 73uteinizi, Lincomycin, Linezolid, Lomefloxacin, Loracarbef, Meropenem, Metronidazole, Minocycline, Moxifloxacin, Nafcillin, Nalidixic Acid, Neomycin, Norfloxacin, Ofloxacin, Omadacycline, Oritavancin, Oxacillin, Oxytetracycline, Paromomycin, Penicillin, Pentamidine, Piperacillin, Plazomicin, Quinupristin, Rifaximin, Sarecycline, Secnidazole, Sparfloxacin, Spectinomycin, Sulfamethoxazole, Sulfisoxazole, Tedizolid, Telavancin, Telithromycin, Ticarcillin, Tigecycline, Tobramycin, Trimethoprim, Trovafloxacin, and Vancomycin.

[0442] Representative examples of antiviral agents include, but are not limited to, abacavir, acyclovir, adefovir, amantadine, amprenavir, atazanavir, balavir, baloxavir marboxil, boceprevir, cidofovir, cobicistat, daclatasvir, darunavir, delavirdine, didanosine, docasanol, dolutegravir, doravirine, ecoliever, edoxudine, efavirenz, elvitegravir, emtricitabine, enfuvirtide, entecavir, etravirine, famciclovir, fomivirsen, fosamprenavir, foscarnet, fosnonet, famciclovir, favipravir, fomivirsen, foscavir, ganciclovir, ibacitabine, idoxuridine, indinavir, inosine, inosinepranobex), interferon type I, interferon type II, interferon type III, lamivudine, letermovir, letermovir, lopinavir, loviride, maraviroc, methisazone, moroxydine, nelfinavir, nevirapine, nitazoxanide, oseltamivir, pegylated interferon alpha-2a, pegylated interferon alpha-2b, penciclovir, peramivir, pleconaril, podophyllotoxin, pyrimidine, raltegravir, 73uteinizin, ribavirin, rilpivirine, rimantadine, rintatolimod, molnupiravir, ritonavir, saquinavir, simeprevir, sofosbuvir, stavudine, tarabivirin, telaprevir, telbivudine, tenofovir alafenamide, tenofovir disoproxil, tenofovir, tipranavir, trifluridine, trizivir, tromantadine, umifenovir, valaciclovir, valganciclovir, vidarabine, zalcitabine, zanamivir, and zidovudine.

[0443] Representative examples of anticoagulants include, but are not limited to, heparin, warfarin, rivaroxaban, dabigatran, apixaban, edoxaban, enoxaparin, and fondaparinux.

[0444] Representative examples of antiplatelet agents include, but are not limited to, clopidogrel, ticagrelor, prasugrel, dipyridamole, dipyridamole / aspirin, ticlopidine, and eptifibatide.

[0445] Representative examples of antifungal agents include, but are not limited to, voriconazole, itraconazole, 74uteinizing 74, fluconazole, ketoconazole, clotrimazole, isavuconazonium, miconazole, caspofungin, anidulafungin, micafungin, griseofulvin, terbinafine, flucytosine, terbinafine, nystatin, and amphotericin b.

[0446] Representative examples of steroidal anti-inflammatory agents include, but are not limited to, hydrocortisone, dexamethasone, prednisolone, prednisone, triamcinolone, methylprednisolone, budesonide, betamethasone, cortisone, and deflazacort. Representative examples of non-steroidal anti-inflammatory drugs include ibuprofen, naproxen, ketoprofen, tolmetin, etodolac, fenoprofen, flurbiprofen, diclofenac, piroxicam, indomethacin, sulindax, meloxicam, nabumetone, oxaprozin, mefenamic acid, and diflunisal.

[0447] Other examples of active agents include chloroquine, hydroxychloroquine, pyridoxal phosphate, vitamin D, and vitamin C.

[0448] Representative examples of anti-cytokine or immunomodulatory agents include, but are not limited to, tocilizumab, salinomycin, bevacizumab, fingolimod, imiquimod, and eculizumab.

[0449] Immunotherapeutic agents can include, but are not limited to, anti-CD40 antibodies, anti-PDL1 antibodies (e.g., atezolizumab, durvalumab, or avelumab), anti-PD1 antibodies, anti-CTLA4 antibodies, programmed death protein 1 (PD-1) inhibitors, or programmed death protein ligand 1 or 2 inhibitors including (e.g., nivolumab (BMS), pembrolizumab (Merck), pidilizumab (CureTech / Teva), AMP-244 (Amplimmune / GSK), BMS-936559 (BMS), and MEDI4736 (Roche / Genentech)) or combinations thereof.

[0450] Representative examples of contraceptives include, but are not limited to, progestogens, estrogens, or any combination thereof. For example, suitable progestogens include, but are not limited to, natural and synthetic compounds having progestational activity, such as, for example, progesterone, chlormadinone acetate, norethisterone, cyproterone acetate, norethynodrel, desogestrel, levonorgestrel, drospirenone, trimegestone, norgestrel, norgestimate, megestrol acetate, etonogestrel, gestodene, and other natural and / or synthetic progestogens. For example, suitable estrogens include, but are not limited to, natural and synthetic compounds having estrogenic activity, such as estradiol (17β-estradiol), 17α-estradiol, estriol, estrone, and their esters, such as the acetate, sulfate, valerate, or benzoate esters of these compounds, including, for example, estradiol 17β-cypionate, estradiol 17-propionate, estradiol 3-benzoate, and piperazine estrone sulfate; ethinyl estradiol; conjugated estrogens (natural and synthetic); mestranol; agonist antiestrogens; and selective estrogen receptor modulators. Other examples of contraceptives include luteinizing hormone (GnRh) or analogs thereof, such as deslorelin, avorelin, leuprolide, triptorelin, nafarelin, goserelin, buserelin, and fertirelin.

[0451] The term "steroid" refers to compounds belonging to or related to the following exemplary families of compounds: corticosteroids, mineralocorticoids, and sex steroids (including, for example, potential androgens or estrogens and antiandrogen and antiestrogen molecules). These include, for example, prednisone, prednisolone, methylprednisolone, triamcinolone, fluocinolone acetonide, aldosterone, spironolactone, danazol (also known as OPTINA), and others. In some embodiments, the therapeutic agent can comprise a steroid.

[0452] Exemplary cancer drugs or anti-cancer agents can include, but are not limited to, antimetabolite anti-cancer agents and anti-mitotic anti-cancer agents and combinations thereof. A variety of antimetabolites and anti-mitotic anti-cancer agents (including a single such agent or combinations of such agents) can be used in the methods and compositions described herein.

[0453] Antimetabolite anti-cancer agents are generally structurally similar to natural metabolites that are involved in normal metabolic processes in cancer cells, such as the synthesis of nucleic acids and proteins. However, the antimetabolites are sufficiently different from the natural metabolites such that they interfere with the metabolic processes in cancer cells. In cells, the antimetabolites are mistaken for their similar metabolites and are processed by the cells in a manner similar to the normal compounds. The presence of the "decoy" metabolites prevents the cells from performing important functions, and the cells cannot grow and survive. For example, antimetabolites can exert cytotoxic activity by incorporating these fraudulent nucleotides into cellular DNA, thereby disrupting cell division, or by inhibiting key cellular enzymes that prevent DNA replication.

[0454] Thus, in one aspect, the antimetabolite anti-cancer agents are nucleotides or nucleotide analogs. In certain aspects, for example, the antimetabolite can comprise a purine (e.g., guanine or adenosine) or an analog thereof, or a pyrimidine (cytidine or thymidine) or an analog thereof, with or without a linked sugar moiety.

[0455] Suitable antimetabolite anti-cancer agents for the present disclosure can generally be classified according to the metabolic processes they affect and can include, but are not limited to, analogs and derivatives of folic acid, pyrimidines, purines, and cytidine. Thus, in one aspect, the antimetabolites are selected from the group consisting of cytidine analogs, folic acid analogs, purine analogs, pyrimidine analogs, and combinations thereof.

[0456] In a particular aspect, for example, the antimetabolite is a cytidine analog. According to this aspect, for example, the cytidine analog can be selected from the group consisting of cytarabine (cytosine arabinoside), 76uteinizing (5-azacytidine) and its salts, analogs, and derivatives.

[0457] In another particular aspect, for example, the antimetabolite is a folic acid analog. Folic acid analogs or antifolates generally act by inhibiting dihydrofolate reductase (DHFR), an enzyme involved in nucleotide formation; when this enzyme is blocked, nucleotides are not formed, thereby disrupting DNA replication and cell division. According to certain aspects, for example, the folic acid analog can be selected from the group consisting of demethylmethotrexate, methotrexate (amethopterin), pemetrexed, pteropterin, raltitrexed, trimetrexate, and their salts, analogs, and derivatives.

[0458] In another specific aspect, for example, the antimetabolite is a purine analogue. Purine-based antimetabolites act by inhibiting DNA synthesis, for example, by interfering with the production of purines containing nucleotides, adenine, and guanine, thereby stopping DNA synthesis and thus cell division. Purine analogues can also be incorporated into the DNA molecule itself during DNA synthesis, which can interfere with cell division. According to certain aspects, for example, the purine analogue can be selected from the group consisting of: acyclovir, allopurinol, 2-aminoadenosine, vidarabine (ara-A), luteinizing, luteinizing 76, 8-aza-adenosine, 8-fluoro-adenosine, 8-methoxy-adenosine, 8-oxo-adenosine, cladribine, deoxycoformycin, fludarabine, ganciclovir, 8-aza-guanosine, 8-fluoro-guanosine, 8-methoxy-guanosine, 8-oxo-guanosine, guanosine diphosphate, guanosine diphosphate-β-L-2-amino fucose, guanosine diphosphate-D-arabinose, guanosine diphosphate-2-fluoro fucose, guanosine diphosphate fucose, mercaptopurine (6-MP), pentostatin, tioguanine, thioguanine (6-TG), and their salts, analogues, and derivatives.

[0459] In another specific aspect, for example, the antimetabolite is a pyrimidine analogue. Similar to the purine analogues discussed above, pyrimidine-based antimetabolites block the synthesis of pyrimidine nucleotides (cytosine and thymine in DNA; cytosine and uracil in RNA). By acting as a "decoy", pyrimidine-based compounds can prevent the production of nucleotides and / or can be incorporated into the growing DNA strand and cause its termination. According to certain aspects, for example, the pyrimidine analogue can be selected from the group consisting of: ancitabine, luteinizing, 6-azauridine, bromouracil (e.g., 5-bromouracil), capecitabine, carmofur, chlorouracil (e.g., 5-chlorouracil), cytarabine (cytosine arabinoside), cytosine, didoxuridine, 3'-azido-3'-deoxythymidine, 3'-luteinizing 77ine-2'-ene, 3'-deoxy-3'-deoxythymidine-2'-ene, dihydrouracil, doxifluridine, enocitabine, floxuridine, 5-fluorocytosine, 2-fluorodeoxycytidine, 3-fluoro-3'-deoxythymidine, fluorouracil (e.g., 5-fluorouracil (also known as 5-FU)), gemcitabine, 5-methylcytosine, 5-propynylcytosine, 5-propynylthymine, 5-propynyluracil, thymine, uracil, uridine, and their salts, analogues, and derivatives. In one aspect, the pyrimidine analogue is not 5-fluorouracil. In another aspect, the pyrimidine analogue is gemcitabine or its salt.

[0460] In some aspects, the antimetabolite is selected from the group consisting of 5-fluorouracil, capecitabine, 6-mercaptopurine, methotrexate, gemcitabine, cytarabine, fludarabine, pemetrexed and its salts, analogs, derivatives and combinations. In other aspects, the antimetabolite is selected from the group consisting of capecitabine, 6-mercaptopurine, methotrexate, gemcitabine, cytarabine, fludarabine, pemetrexed and its salts, analogs, derivatives and combinations. In a particular aspect, the antimetabolite is not 5-fluorouracil. In a particularly preferred aspect, the antimetabolite is gemcitabine or its salt (such as gemcitabine hydrochloride ).

[0461] Other antimetabolite anticancer agents may be selected from, but are not limited to, the group consisting of acivicin, aminothiadiazole, brequinar sodium, Ciba-Geigy CGP-30694, cyclopentyl cytosine, cytarabine stearate phosphate, cytarabine conjugate, Lilly DATHF, MerrelDow DDFC, dideoxyguanosine, didox, Yoshitomi DMDC, Wellcome EHNA, Merck&Co.EX-015, fazarabine, fludarabine phosphate, N-(2′-furanylalkyl)-5-fluorouracil, Daiichi Seiyaku FO-152, 5-FU-fibrinogen, isopropyl pyrrolizine, Lilly LY-188011; Lilly LY-264618, topramezone, Wellcome MZPES, norspermidine, NCI NSC-127716, NCI NSC-264880, NCI NSC-39661, NCI NSC-612567, Warner-Lambert PALA, pentostatin, pirroxantrone, plicamycin, Asahi Chemical PL-AC, Takeda TAC-788, thiazofurin, Erbamont TIF, tyrosine kinase inhibitor, Taiho UFT and uricytin, etc.

[0462] In one aspect, the antimitotic anticancer agent is a microtubule inhibitor or a microtubule stabilizer. Generally, microtubule stabilizers, such as taxanes and epothilones, bind to the inner surface of the β-tubulin chain and enhance microtubule assembly by promoting the nucleation and elongation phases of the polymerization reaction and by reducing the critical tubulin subunit concentration required for microtubule assembly. Different from microtubule inhibitors (such as vinca alkaloids) that hinder microtubule assembly, microtubule stabilizers (such as taxanes) reduce the lag time and significantly shift the dynamic equilibrium between tubulin dimers and microtubule polymers towards the polymerization direction. Thus, in one aspect, the microtubule stabilizer is a taxane or an epothilone. In another aspect, the microtubule inhibitor is a vinca alkaloid.

[0463] In some embodiments, the anti-cancer agent can comprise a taxane or its derivatives or analogs. The taxane can be a naturally derived compound or related form, or can be a chemically synthesized compound or its derivative, having anti-tumor properties. Taxanes are a family of terpenoids, including but not limited to paclitaxel and docetaxel which are mainly derived from the Pacific yew tree Taxus brevifolia, and have activity against certain tumors, particularly breast and ovarian tumors. In one aspect, the taxane is docetaxel or paclitaxel. Paclitaxel is the preferred taxane and is considered an anti-mitotic agent that promotes the assembly of tubulin dimers into microtubules and stabilizes the microtubules by preventing depolymerization. This stability results in the inhibition of the normal dynamic reorganization of the microtubule network, which is essential for interphase and mitotic cell functions.

[0464] Also included are various known taxane derivatives, including hydrophilic derivatives and hydrophobic derivatives. Taxane derivatives include, but are not limited to, galactose and mannose derivatives described in International Patent Application No. WO 99 / 18113; piperazine and other derivatives described in WO99 / 14209; taxane derivatives described in WO 99 / 09021, WO 98 / 22451, and U.S. Patent No. 5,869,680; 6-thio derivatives described in WO 98 / 28288; sulfinamide derivatives described in U.S. Patent No. 5,821,263; deoxytaxol compounds such as those described in U.S. Patent No. 5,440,056; and tacozolomide derivatives described in U.S. Patent No. 5,415,869. As described above, it further includes prodrugs of paclitaxel, including, but not limited to, those described in WO 98 / 58927; WO 98 / 13059; and U.S. Patent No. 5,824,701. The taxane can also be a taxane conjugate, such as, for example, paclitaxel-PEG, paclitaxel-dextran, paclitaxel-xylose, docetaxel-PEG, docetaxel-dextran, docetaxel-xylose, etc. Other derivatives are mentioned in references such as “Synthesis and Anticancer Activity of Taxol Derivatives,” D.G.I. Kingston et al., Studies in Organic Chemistry, Vol. 26, entitled “New Trends in Natural Products Chemistry” (1986), Atta-ur-Rabman, P.W. le Quesne, editors (Elsevier, Amsterdam 1986). Each of these references is hereby incorporated by reference in its entirety into this text.

[0465] The various taxanes can be readily prepared using techniques known to those of skill in the art (see also WO 94 / 07882, WO 94 / 07881, WO 94 / 07880, WO 94 / 07876, WO 93 / 23555, WO 93 / 10076; U.S. Patent Nos. 5,294,637; 5,283,253; 5,279,949; 5,274,137; 5,202,448; 5,200,534; 5,229,529; and EP 590,267), each of which is incorporated herein by reference in its entirety, or obtained from a variety of commercial sources, including, for example, Sigma-Aldrich Co., St. Louis, Mo.

[0466] Alternatively, the antimitotic anti-cancer agent can be a microtubule inhibitor; in a preferred aspect, the microtubule inhibitor is a vinca alkaloid. Generally, vinca alkaloids are mitotic spindle poisons. Vinca alkaloid agents act during mitosis, when the chromosomes divide and begin to migrate along the tubules of the mitotic spindle towards one of its poles before cell separation. Under the action of these spindle poisons, the spindle becomes disordered due to the dispersion of chromosomes during mitosis, affecting cell proliferation. According to certain aspects, for example, vinca alkaloids are selected from the group consisting of vinblastine, vincristine, vindesine, vinorelbine, and their salts, analogs, and derivatives.

[0467] The antimitotic anti-cancer agent can also be an epothilone. Generally, members of the epothilone class of compounds stabilize microtubule function according to a mechanism similar to that of taxanes. Epothilones can also cause cell cycle arrest at the G2-M transition phase, leading to cytotoxicity and ultimately apoptosis. Suitable epothilones include epothilone A, epothilone B, epothilone C, epothilone D, epothilone E, and epothilone F, and their salts, analogs, and derivatives. A specific epothilone analog is the epothilone B analog ixempra TM ).

[0468] In certain aspects, the antimitotic anti-cancer agent is selected from the group consisting of taxanes, epothilones, vinca alkaloids, and their salts and combinations. Thus, for example, in one aspect, the antimitotic agent is a taxane. In this regard, more preferably, the antimitotic agent is paclitaxel or docetaxel, still more preferably paclitaxel. In another aspect, the antimitotic agent is an epothilone (e.g., an epothilone B analog). In another aspect, the antimitotic agent is a vinca alkaloid.

[0469] Examples of anti-cancer drugs that can be used in the present disclosure include, but are not limited to: thalidomide; platinum coordination complexes such as cis-DDP, oxaliplatin, and carboplatin; anthraquinones such as mitoxantrone; substituted ureas such as hydroxyurea; methylhydrazine derivatives such as procarbazine (N-methylhydrazine, MIH); adrenocortical inhibitors such as mitotane (o,p'-DDD) and aminoglutethimide; RXR agonists such as bexarotene; and tyrosine kinase inhibitors such as sunitinib and imatinib.

[0470] Examples of additional anti-cancer drugs include alkylating agents, antimetabolites, natural products, hormones and antagonists, and other drugs. Alternative names are indicated in parentheses. Examples of alkylating agents include nitrogen mustards such as dichloromethyldiethylamine, cyclophosphamide, ifosfamide, melphalan (sarcolysin), and chlorambucil; ethyleneimines and methylmelamines such as altretamine and thiotepa; alkyl sulfonates such as busulfan; nitrosoureas such as carmustine (BCNU), semustine (methyl-CCNU), lomustine (CCNU), and streptozocin (streptozotocin); DNA synthesis antagonists such as estramustine phosphate; and triazines such as dacarbazine (DTIC, dimethyl-triazenoimidazole carboxamide) and temozolomide. Examples of antimetabolites include folic acid analogs such as methotrexate; pyrimidine analogs such as fluorouracil (5-fluorouracil, 5-FU, SFU), floxuridine (fluorodeoxyuridine, FudR), cytarabine (cytosine arabinoside), and gemcitabine; purine analogs such as mercaptopurine (6-mercaptopurine, 6-MP), thioguanine (6-thioguanine, TG), and pentostatin (2'-deoxycoformycin, deoxycoformycin), cladribine, and fludarabine; and topoisomerase inhibitors such as amsacrine. Examples of natural products include vinca alkaloids such as vinblastine (VLB) and vincristine; taxanes such as paclitaxel, albumin-bound paclitaxel (Abraxane), and docetaxel (Taxotere); epipodophyllotoxins such as etoposide and teniposide; camptothecins such as topotecan and irinotecan; antibiotics such as dactinomycin (actinomycin D), daunorubicin (daunomycin, rubidomycin), doxorubicin, histrelin, bleomycin, mitomycin (mitomycin C), idarubicin, epirubicin; enzymes such as L-asparaginase; and biological response modifiers such as interferon α and interleukin 2.Examples of hormones and antagonists include luteinizing hormone releasing hormone agonists such as buserelin; adrenocorticosteroids such as prednisone and related preparations; progesterones such as hydroxyprogesterone caproate, medroxyprogesterone acetate, and megestrol acetate; estrogens such as diethylstilbestrol, ethinyl estradiol, and related preparations; estrogen antagonists such as tamoxifen and anastrozole; androgens such as testosterone propionate, fluoxymesterone, and related preparations; androgen antagonists such as flutamide and bicalutamide; and gonadotropin releasing hormone analogs such as leuprolide. Alternative names and trade names of these and additional examples of cancer drugs, as well as their methods of use (including dosing and administration regimens), will be known to those of skill in the art.

[0471] In some aspects, the anti-cancer agent can comprise a chemotherapeutic agent. Suitable chemotherapeutic agents include, but are not limited to, alkylating agents, antibiotic agents, antimetabolites, hormonal agents, plant-derived agents and their synthetic derivatives, anti-angiogenic agents, differentiation inducers, cell growth arrest inducers, apoptosis inducers, cytotoxic agents, agents that affect cellular bioenergetics (i.e., agents that affect cellular ATP levels and molecules / activities that regulate these levels), biological agents (e.g., monoclonal antibodies, kinase inhibitors, and inhibitors of growth factors and their receptors), gene therapy agents, cell therapy (e.g., stem cells), or any combination thereof.

[0472] According to these aspects, the chemotherapeutic agent is selected from the group consisting of cyclophosphamide, chlorambucil, melphalan, nitrogen mustard, ifosfamide, busulfan, lomustine, streptozocin, temozolomide, dacarbazine, cisplatin, carboplatin, oxaliplatin, procarbazine, uracil mustard, methotrexate, pemetrexed, fludarabine, cytarabine, fluorouracil, floxuridine, gemcitabine, capecitabine, vinblastine, vincristine, vinorelbine, etoposide, paclitaxel, docetaxel, doxorubicin, daunorubicin, epirubicin, idarubicin, mitoxantrone, bleomycin, mitomycin, hydroxyurea, topotecan, irinotecan, amsacrine, teniposide, erlotinib hydrochloride, and combinations thereof. Each possibility represents an independent aspect of the present invention.

[0473] The anti-tumor agent can be selected from the group consisting of: Abiraterone Acetate, Abitrexate (Methotrexate), Abraxane (Paclitaxel Albumin-stabilized Nanoparticle Formulation), ABVD, ABVE, ABVE-PC, AC, AC-T, Adcetris (Brentuximab Vedotin), ADE, Ado-Trastuzumab Emtansine, Adriamycin (Doxorubicin Hydrochloride), Adrucil (Fluorouracil), Afatinib Dimaleate, Afinitor (Everolimus), Akynzeo (Netupitant and Palonosetron Hydrochloride), Aldara (Imiquimod), Aldesleukin, Alemtuzumab, Alimta (Pemetrexed Disodium), Aloxi (Palonosetron Hydrochloride), Ambochlorin (Chlorambucil), Amboclorin (Chlorambucil), Aminolevulinic Acid, Anastrozole, Aprepitant, Aredia (Pamidronate Disodium), Arimidex (Anastrozole), Aromasin (Exemestane), Arranon (Nelarabine), Arsenic Trioxide, Arzerra (Ofatumumab), AsparaginaseErwinia chrysanthemi), Avastin (Bevacizumab), Axitinib, Azacitidine, BEACOPP, Becenum (Carmustine), Beleodaq (Belinostat), Belinostat, Bendamustine Hydrochloride, BEP, Bevacizumab, Bexarotene, Bexxar (Tositumomab and Iodine I 131 Tositumomab), Bicalutamide, BiCNU (Carmustine), Bleomycin, Blinatumomab, Blincyto (Blinatumomab), Bortezomib, Bosulif (Bosutinib), Bosutinib, Brentuximab Vedotin, Busulfan, Busulfex (Busulfan), Cabazitaxel, Cabozantinib-S-Malate, CAF, Campath (Alemtuzumab), Camptosar (Irinotecan Hydrochloride), Capecitabine, CAPOX, Carboplatin, Carboplatin-Taxol, Carfilzomib, Carmubris (Carmustine), Carmustine, Carmustine Implant, Casodex (Bicalutamide), CeeNU (Lomustine), Ceritinib, Cerubidine (DaunorubicinHydrochloride)), Cervarix (Recombinant Bivalent HPV Vaccine), Cetuximab, Chlorambucil, Chlorambucil - Prednisone, CHOP, Cisplatin, Clafen (Cyclophosphamide), Clofarabine, Clofarex (Clofarabine), Clolar (Clofarabine), CMF, Cometriq (Cabozantinib Malate), COPP, COPP - ABV, Cosmegen (Dactinomycin), Crizotinib, CVP, Cyclophosphamide, Cyfos (Ifosfamide), Cyramza (Ramucirumab), Cytarabine, Cytarabine, Liposome, Cytosar - U (Cytarabine), Cytoxan (Cyclophosphamide), Dabrafenib, Dacarbazine, Dacogen (Decitabine), Dactinomycin, Dasatinib, Daunorubicin Hydrochloride, Decitabine, Degarelix, Denileukin Diftitox, Denosumab, DepoCyt (Liposomal Cytarabine), DepoFoam (Liposomal Cytarabine), Dexrazoxane Hydrochloride, Dinutuximab, Docetaxel, Doxil (Liposomal Doxorubicin Hydrochloride), Doxorubicin Hydrochloride, Liposomal Doxorubicin Hydrochloride, Dox - SL (Liposomal Doxorubicin Hydrochloride), DTIC - Dome (Dacarbazine), Efudex (Fluorouracil), Elitek (Rasburicase), Ellence (Epirubicin HydrochlorideHydrochloride)), Eloxatin (Oxaliplatin), Eltrombopag Olamine, Emend (Aprepitant), Enzalutamide, Epirubicin Hydrochloride, EPOCH, Erbitux (Cetuximab), Eribulin Mesylate, Erivedge (Vismodegib), Erlotinib Hydrochloride, Erwinaze (Aspergillus oryzae asparaginase), Etopophos (Etoposide Phosphate), Etoposide, Etoposide Phosphate, Evacet (Doxorubicin Hydrochloride Liposome), Everolimus, Evista (Raloxifene Hydrochloride), Exemestane, Fareston (Toremifene), Farydak (Panobinostat), Faslodex (Fulvestrant), FEC, Femara (Letrozole), Filgrastim, Fludara (Fludarabine Phosphate), Fludarabine Phosphate, Fluoroplex (Fluorouracil), Fluorouracil, Folex (Methotrexate), Folex PFS (Methotrexate), FOLFIRI, FOLFIRI - Bevacizumab, FOLFIRI - Cetuximab, FOLFIRINOX, FOLFOX, Folotyn (Pralatrexate), FU - LV, Fulvestrant, Gardasil (Recombinant Human Papillomavirus Quadrivalent Vaccine), Gardasil 9 (Recombinant Human Papillomavirus Nonavalent Vaccine), Gazyva (Obinutuzumab), Gefitinib, Gemcitabine HydrochlorideHydrochloride), Gemcitabine-Cisplatin, Gemcitabine-Oxaliplatin, Gemtuzumab Ozogamicin, Gemzar (Gemcitabine Hydrochloride), Gilotrif (Afatinib Dimaleate), Gleevec (Imatinib Mesylate), Gliadel (Carmustine Implant), Gliadel wafer (Carmustine Implant), Glucarpidase, Goserelin Acetate, Halaven (Eribulin Mesylate), Herceptin (Trastuzumab), Recombinant Bivalent HPV Vaccine, Recombinant Nonavalent HPV Vaccine, Recombinant Quadrivalent HPV Vaccine, Hycamtin (Topotecan Hydrochloride), Hyper-CVAD, Ibrance (Palbociclib), Ibritumomab Tiuxetan, Ibrutinib, ICE, Iclusig (Ponatinib Hydrochloride), Idamycin (Idarubicin Hydrochloride), Idarubicin Hydrochloride, Idelalisib, Ifex (Ifosfamide), Ifosfamide, Ifosfamidum (Ifosfamide), Imatinib Mesylate, Imbruvica (Ibrutinib), Imiquimod, Inlyta (Axitinib), Interferon α-2b, Recombinant Interferon α-2b with Intron A, Iodine I 131 Tositumomab and Tositumomab, Ipilimumab, Iressa (Gefitinib), Irinotecan Hydrochloride, Istodax (Romidepsin), Ixabepilone, Ixempra (Ixabepilone), Jakafi (RuxolitinibPhosphate)), Jevtana (Cabazitaxel), Kadcyla (Ado-Trastuzumab Emtansine), Keoxifene (Raloxifene Hydrochloride), Kepivance (Palifermin), Keytruda (Pembrolizumab), Kyprolis (Carfilzomib), Lanreotide Acetate, Lapatinib Ditosylate, Lenalidomide, Lenvatinib Mesylate, Lenvima (Lenvatinib Mesylate), Letrozole, Leucovorin Calcium, Leukeran (Chlorambucil), Leuprolide Acetate, Levulan (Aminolevulinic Acid), Linfolizin (Chlorambucil), LipoDox (Doxorubicin Hydrochloride Liposome), Liposomal Cytarabine, Lomustine, Lupron (Leuprolide Acetate), Lupron Depot (Leuprolide Acetate), Lupron Depot-Ped (Leuprolide Acetate), Lupron Depot - 3 months (Leuprolide Acetate), Lupron Depot - 4 months (Leuprolide Acetate), Lynparza (Olaparib), Marqibo (Vincristine Sulfate Liposome), Matulane (Procarbazine Hydrochloride), Mechlorethamine Hydrochloride, Megace (Megestrol Acetate), Megestrol Acetate, Mekinist (Trametinib), Mercaptopurine, Mesna, Mesnex (Mesna), Methazolastone (Temozolomide), Methotrexate, Methotrexate LPF (Methotrexate), Mexate (Methotrexate), Mexate - AQ (Methotrexate), Mitomycin C, MitoxantroneHydrochloride), Mitozytrex (Mitomycin C), MOPP, Mozobil (Plerixafor), Mustargen (Mechlorethamine Hydrochloride), Mutamycin (Mitomycin C), Myleran (Busulfan), Mylosar (Azacitidine), Mylotarg (Gemtuzumab Ozogamicin), Nanoparticle Paclitaxel (Paclitaxel Albumin-Stabilized Nanoparticle Formulation), Navelbine (Vinorelbine Tartrate), Nelarabine, Neosar (Cyclophosphamide), Netupitant and Palonosetron Hydrochloride, Neupogen (Filgrastim), Nexavar (Sorafenib Tosylate), Nilotinib, Nivolumab, Nolvadex (Tamoxifen Citrate), Nplate (Romiplostim), Obinutuzumab, Odomzo (Sonidegib), OEPA, Ofatumumab, OFF, Olaparib, Omacetaxine Mepesuccinate, Oncaspar (Pegaspargase), Ondansetron HydrochlorideHydrochloride, Ontak (Denileukin Diftitox), Opdivo (Nivolumab), OPPA, Oxaliplatin, Paclitaxel, Paclitaxel Albumin-Stabilized Nanoparticle Formulation, PAD, Palbociclib, Palifermin, Palonosetron Hydrochloride, Palonosetron Hydrochloride and Netupitant, Pamidronate Disodium, Panitumumab, Panobinostat, Paraplat (Carboplatin), Paraplatin (Carboplatin), Pazopanib Hydrochloride, Pegaspargase, Peginterferon Alfa-2b, PEG-Intron (Peginterferon Alfa-2b), Pembrolizumab, Pemetrexed Disodium, Perjeta (Pertuzumab), Pertuzumab, Platinol (Cisplatin), Platinol-AQ (Cisplatin), Plerixafor, Pomalidomide, Pomalyst (Pomalidomide), Ponatinib Hydrochloride, Pralatrexate, Prednisone, Procarbazine Hydrochloride, Proleukin (Aldesleukin), Prolia (Denosumab), Promacta (Eltrombopag Olamine), Provenge (Sipuleucel-T), Purinethol (Mercaptopurine), Purixan (Mercaptopurine), Radium 223 Dichloride, Raloxifene Hydrochloride, Ramucirumab, Rasburicase, R-CHOP, R-CVP, Recombinant Human Papillomavirus (HPV) Bivalent Vaccine, Recombinant Human Papillomavirus (HPV) 9-Valent Vaccine, Recombinant Human Papillomavirus (HPV) Quadrivalent Vaccine, Recombinant Interferon Alfa-2b, Regorafenib, R-EPOCH, Revlimid (Lenalidomide), Rheumatrex (Methotrexate), Rituxan (Rituximab), Rituximab, Romidepsin, Romiplostim, Rubidomycin (Daunorubicin Hydrochloride), Ruxolitinib Phosphate, Sclerosol Intrapleural Aerosol (Talc), Siltuximab, Sipuleucel-T, Somatuline Depot (Lanreotide Acetate), Sonidegib, Sorafenib Tosylate, Sprycel (Dasatinib), STANFORDV, Sterile Talc (talc), Steritalc (talc), Stivarga (regorafenib), Sunitinib Malate, Sutent (sunitinib malate), Sylatron (peginterferon alfa-2b), Sylvant (siltuximab), Synovir (thalidomide), Synribo (homoharringtonine), TAC, Tafinlar (dabrafenib), Talc, Tamoxifen Citrate, Tarabine PFS (cytarabine), Tarceva (erlotinib hydrochloride), Targretin (bexarotene), Tasigna (nilotinib), Taxol (paclitaxel), Taxotere (docetaxel), Temodar (temozolomide), Temozolomide, Temsirolimus, Thalidomide, Thalomid (thalidomide), Thiotepa, Toposar (etoposide), Topotecan Hydrochloride, Toremifene, Torisel (temsirolimus), Tositumomab and Iodine I 131 Tositumomab, Totect (dexrazoxane hydrochloride), TPF, Trametinib, Trastuzumab, Treanda (bendamustine hydrochloride), Trisenox (arsenic trioxide), Tykerb (lapatinib ditosylate), Unituxin (dinutuximab), Vandetanib, VAMP, Vectibix (panitumumab), VeIP, Velban (vinblastine sulfate), Velcade (bortezomib), Velsar (vinblastine sulfate), Vemurafenib, VePesid (etoposide), Viadur (leuprolide acetate), Vidaza (azacitidine), Vinblastine Sulfate, Vincasar PFS (vincristinePFS) (Vincristine Sulfate), vincristine sulfate, vincristine sulfate liposome, vinorelbine tartrate, VIP, vismodegib, Voraxaze (carboxypeptidase), Vorinostat, Votrient (pazopanib hydrochloride), Wellcovorin (calcium folinate), Xalkori (crizotinib), Xeloda (capecitabine), XELIRI, XELOX, Xgeva (denosumab), Xofigo (radium-223 dichloride), Xtandi (enzalutamide), Yervoy (ipilimumab), Zaltrap (Ziv-Aflibercept), Zelboraf (vemurafenib), Zevalin (ibritumomab tiuxetan), Zinecard (dexrazoxane hydrochloride), Zofran (ondansetron hydrochloride), Zoladex (goserelin acetate), Zoledronic Acid, Zolinza (vorinostat), Zometa (zoledronic acid), Zydelig (idelalisib), Zykadia (ceritinib), and Zytiga (abiraterone acetate).

[0474] Growth factors that can be used as therapeutic agents include, but are not limited to, transforming growth factor-α (“TGF-α”), transforming growth factor (“TGF-β”), platelet-derived growth factor (“PDGF”), fibroblast growth factor (“FGF”) (including FGF acidic isomers 1 and 2), FGF basic form 2 and FGF 4, 8, 9, and 10, nerve growth factor (“NGF”) (including NGF 2.5s, NGF7.0s, and βNGF and neurotrophins), brain-derived neurotrophic factor, cartilage-derived factor, bone growth factor (BGF), basic fibroblast growth factor, insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF), granulocyte colony-stimulating factor (G-CSF), insulin-like growth factor (IGF) I and II, hepatocyte growth factor, glial cell line-derived neurotrophic factor (GDNF), stem cell factor (SCF), keratinocyte growth factor (KGF), transforming growth factor (TGF) (including TGFα, β, β1, β2, β3), skeletal growth factor, bone matrix-derived growth factor, and bone-derived growth factor, and mixtures thereof.

[0475] Immunoglobulins useful in the present disclosure include, but are not limited to, IgG, IgA, IgM, IgD, IgE, and mixtures thereof. Some preferred growth factors include VEGF (vascular endothelial growth factor), NGF (nerve growth factor), PDGF-AA, PDGF-BB, PDGF-AB, FGFb, FGFa, and BGF.

[0476] Other molecules useful as anti-cancer agents include, but are not limited to, growth hormone, leptin, leukemia inhibitory factor (LIF), tumor necrosis factor α and β, endostatin, thrombospondin, osteogenic protein-1, bone morphogenetic proteins 2 and 7, osteonectin, somatomedin-like peptides, and osteocalcin.

[0477] Tumor antigens can be based on specific mutations (neoepitopes) and those mutations in cancer germline gene expression (antigens shared by tumors found in multiple patients, referred to herein as "conventional cancer antigens" or "shared cancer antigens"). In some embodiments, conventional antigens are antigens known to be prevalent in cancers or tumors or in specific types of cancers or tumors. In some embodiments, conventional cancer antigens are non-mutated tumor antigens. In some embodiments, conventional cancer antigens are mutated tumor antigens.

[0478] Diagnostic agents include gases; metals; commercially available imaging agents for positron emission tomography (PET), computerized axial tomography (CAT), single photon emission computed tomography, x-rays, fluoroscopy, and magnetic resonance imaging (MRI); and contrast agents. Examples of suitable materials for use as contrast agents in MRI include gadolinium chelates, and iron, magnesium, manganese, copper, and chromium. Examples of materials for use in CAT and x-ray imaging include iodine-based materials.

[0479] Vaccines can include isolated proteins or peptides, inactivated organisms and viruses, killed organisms and viruses, genetically altered organisms or viruses, cell extracts, and RNA encoding at least one antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of inducing an immune response against the antigenic polypeptide). The active agent can be combined with interleukins, interferons, cytokines, and adjuvants such as cholera toxin, alum, Freund's adjuvant, etc. The prophylactic agent can include infectious agents such as antigens of bacterial organisms such as Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyogenes, Corynebacterium diphtheriae, Listeria monocytogenes, Bacillus anthracis, Clostridium tetani, Clostridium botulinum, Clostridium perfringens, Neisseria meningitidis, Neisseria gonorrhoeae, Streptococcus mutans, Pseudomonas aeruginosa, Salmonella typhi, Haemophilus parainfluenzae, Bordetella pertussis, Francisella tularensis, Yersinia pestis, Vibrio cholerae, Legionella pneumophila, Mycobacterium tuberculosis, Mycobacterium leprae, Treponema pallidum, Leptospira interrogans, Borrelia burgdorferi, Campylobacter jejuni, etc.;Viruses, such as metapneumoviruses like human metapneumovirus (HMPV), parainfluenza viruses like human parainfluenza virus (HPIV) types 1, 2, and 3 (HPIV1, HPIV2, and HPIV3 respectively), respiratory syncytial virus (RSV), measles virus (MeV), coronaviruses (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, HCoV-HKU1), poxviruses (e.g., smallpox, monkeypox), African swine fever virus, influenza A and B, HIV, varicella-zoster, herpes simplex 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A, B, C, D, and E viruses, etc.; fungi, protozoa, and parasites, such as Cryptococcus neoformans, Histoplasma capsulatum, Candida albicans, Candida tropicalis, Nocardia asteroides, Rickettsia rickettsii, Rickettsia typhi, Mycoplasma pneumoniae, Chlamydia psittaci, Chlamydia trachomatis, Plasmodium falciparum, Trypanosoma brucei, Entamoeba histolytica, Toxoplasma gondii, Trichomonas vaginalis, Schistosoma mansoni, etc. These antigens can be in the form of whole killed organisms, peptides, proteins, glycoproteins, carbohydrates, or combinations thereof.;

[0480] In certain embodiments, the active agent is a nucleic acid. Nucleic acids, polynucleotides, or oligonucleotides that can be introduced according to the methods herein include all types of DNA, cDNA, and RNA sequences. For example, the polynucleotide can be double-stranded DNA, single-stranded DNA, complex DNA, encapsulated DNA, naked RNA, encapsulated RNA, messenger RNA (mRNA), tRNA, short interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), antisense RNA (asRNA), self-amplifying mRNA (saRNA), guide RNA (gRNA), cRNA, and combinations thereof. The polynucleotide can also be a DNA construct, such as an expression vector, an expression vector encoding a desired gene product (e.g., a gene product that is homologous or heterologous to the subject to which it is to be introduced), and the like.

[0481] In some embodiments, nucleic acids (e.g., mRNA) can be used to induce a balanced immune response against an infectious agent. In some embodiments, nucleic acids (e.g., mRNA) can be used to induce a balanced immune response against a virus, such as a paramyxovirus like human metapneumovirus (HMPV), parainfluenza viruses such as human parainfluenza virus (HPIV) types 1, 2, and 3 (HPIV1, HPIV2, and HPIV3, respectively), respiratory syncytial virus (RSV), measles virus (MeV), coronaviruses (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, HCoV-HKU1), poxviruses (e.g., smallpox, monkeypox), African swine fever virus, influenza A and B, HIV, varicella-zoster, herpes simplex 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A, B, C, D, and E viruses, etc. In some embodiments, nucleic acids (e.g., mRNA) can be used to induce a balanced immune response. In some embodiments, nucleic acids (e.g., mRNA) can be used to induce a balanced immune response against respiratory viruses. The term "respiratory virus" as used herein refers to a virus that causes a respiratory disease. For example, negative-sense single-stranded RNA viruses of the family Paramyxoviridae, such as human metapneumovirus (HMPV), human parainfluenza virus (HPIV) types 1, 2, and 3 (HPIV1, HPIV2, and HPIV3, respectively), RSV, and measles virus (MeV). Another example of a respiratory virus is a coronavirus. Metapneumovirus is an enveloped virus with a positive-sense single-stranded RNA genome and a helically symmetric nucleocapsid. Coronaviruses are virus species belonging to the order Nidovirales, family Coronaviridae, and subfamily Orthocoronavirinae.

[0482] Representative examples of betacoronaviruses include, but are not limited to, Embecovirus 1 (e.g., Betacoronavirus 1, Human coronavirus OC43, Chinese mouse coronavirus HKU24, Human coronavirus HKU1, Mouse coronavirus), Hibecovirus (e.g., Bat Hp-betacoronavirus Zhejiang 2013), Merbecovirus (e.g., Hedgehog coronavirus 1, Middle East respiratory syndrome-related coronavirus (MERS-CoV), Pipistrelle bat coronavirus HKU5, Roundleaf bat coronavirus HKU4), Nobecovirus (e.g., Fruit bat coronavirus GCCDC1, Fruit bat coronavirus HKU9), and Sarbecovirus (e.g., Severe acute respiratory syndrome coronavirus (SARS-CoV), Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

[0483] Representative examples of gammacoronaviruses include, but are not limited to, Cegacovirus (e.g., Beluga whale coronavirus SQ1) and Igacovirus (e.g., Avian coronavirus (IBV)).

[0484] Representative examples of deltacoronaviruses include, but are not limited to, Andecovirus (e.g., Mallard coronavirus HKU20), Buldecovirus (e.g., Nightingale coronavirus HKU11, Porcine coronavirus HKU15 (PorCoV HKU15), Java sparrow avian coronavirus HKU13, Japanese white-eye coronavirus HKU16), Herdecovirus (e.g., Night heron coronavirus HKU19), and Moordecovirus (e.g., Common moorhen coronavirus HKU21).

[0485] In some embodiments, the coronavirus is a human coronavirus. Representative examples of human coronaviruses include, but are not limited to, Human coronavirus 229E (HcoV-229E), Human coronavirus OC43 (HcoV-OC43), Human coronavirus HKU1 (HcoV-HKU1), Human coronavirus NL63 (HcoV-NL63), Severe acute respiratory syndrome coronavirus (SARS-CoV), Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and Middle East respiratory syndrome-related coronavirus (MERS-CoV).

[0486] In some embodiments, a nucleic acid (e.g., mRNA) can have an open reading frame that encodes at least one (e.g., at least 2, 3, 4, or 5) human metapneumovirus (Hmpv) antigen polypeptides, human parainfluenza virus (Hpiv) types 1, 2, and 3 (Hpiv1, Hpiv2, and Hpiv3, respectively) antigen polypeptides, respiratory syncytial virus (RSV) antigen polypeptides, measles virus (MeV) antigen polypeptides, varicella-zoster virus antigen polypeptides, influenza virus antigen polypeptides, herpes simplex virus 1 (HSV1) antigen polypeptides, herpes simplex virus 2 (HSV2) antigen polypeptides, poxvirus (e.g., smallpox, monkeypox) antigen polypeptides, African swine fever virus antigen polypeptides, cytomegalovirus antigen polypeptides, Epstein-Barr virus antigen polypeptides, rotavirus antigen polypeptides, rhinovirus antigen polypeptides, adenovirus antigen polypeptides, papillomavirus antigen polypeptides, poliovirus antigen polypeptides, mumps antigen polypeptides, rabies antigen polypeptides, rubella antigen polypeptides, coxsackievirus antigen polypeptides, equine encephalitis antigen polypeptides, Japanese encephalitis antigen polypeptides, yellow fever antigen polypeptides, Rift Valley fever antigen polypeptides, hepatitis A, B, C, D, and E virus antigen polypeptides, or coronavirus (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HcoV-OC43, HcoV-229E, HcoV-NL63, HcoV-NL, HcoV-NH, HcoV-HKU1) antigen polypeptides. As used herein, the term "antigen polypeptide" encompasses immunogenic fragments of antigen polypeptides that induce (or are capable of inducing) an immune response against the following viruses: human metapneumovirus (Hmpv), human parainfluenza virus (Hpiv) types 1, 2, and 3 (Hpiv1, Hpiv2, and Hpiv3, respectively), respiratory syncytial virus (RSV), measles virus (MeV), varicella-zoster, influenza virus, herpes simplex virus type 1 (HSV1), herpes simplex virus type 2 (HSV2), poxvirus (e.g., smallpox, monkeypox), African swine fever virus, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A, B, C, D, and E virus, or coronavirus (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HcoV-OC43, HcoV-229E, HcoV-NL63, HcoV-NL, HcoV-NH, HcoV-HKU1) or any combination thereof.

[0487] In some embodiments, for example, the agent is a nucleic acid (e.g., mRNA), which can induce a balanced immune response against Hmpv, PIV, RSV, MeV, and / or coronavirus (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HcoV-OC43, HcoV-229E, HcoV-NL63, HcoV-NL, HcoV-NH, and / or HcoV-HKU1), or any combination of two or more of the foregoing viruses, comprising both cellular and humoral immunity, without the risk of the possibility of insertional mutagenesis.

[0488] In some embodiments, the nucleic acids disclosed herein can include at least one chemically modified nucleotide. In some embodiments, the at least one chemically modified nucleotide comprises a chemically modified nucleobase, a chemically modified ribose, a chemically modified phosphodiester bond, or a combination thereof.

[0489] In one embodiment, the at least one chemically modified nucleotide is a chemically modified nucleobase.

[0490] In one embodiment, the chemically modified nucleobase is selected from 5-formylcytidine (5Fc), 5-methylcytidine (5meC), 5-methoxycytidine (5moC), 5-hydroxycytidine (5hoC), 5-hydroxymethylcytidine (5hmC), 5-formyluridine (5Fu), 5-methyluridine (5-meU), 5-methoxyuridine (5moU), 5-carboxymethylesteruridine (5camU), pseudouridine (Ψ), N 1 -methylpseudouridine (me 1 Ψ), N 6 -methyladenosine (me 6 A), or thiophenoguanosine ( th G).

[0491] In some embodiments, the chemically modified nucleobase is 5-methoxyuridine (5moU). In some embodiments, the chemically modified nucleobase is pseudouridine (Ψ). In some embodiments, the chemically modified nucleobase is N 1 -methylpseudouridine (me 1 Ψ).

[0492] The structures of these modified nucleobases are shown below:

[0493]

[0494] In one embodiment, the at least one chemically modified nucleotide is a chemically modified ribose.

[0495] In one embodiment, the chemically modified ribose is selected from 2'-O-methyl (2'-O-Me), 2'-fluoro (2'-F), 2'-deoxy-2'-fluoro-β-D-arabinonucleic acid (2'F-ANA), 4'-S, 4'-SFANA, 2'-azido, UNA, 2'-O-methoxy-ethyl (2'-O-ME), 2'-O-allyl, 2'-O-ethylamine, 2'-O-cyanoethyl, locked nucleic acid (LAN), methylene-Clan, N-MeO-amino BNA or N-MeO-aminooxy BNA. In one embodiment, the chemically modified ribose is 2'-O-methyl (2'-O-Me). In one embodiment, the chemically modified ribose is 2'-fluoro (2'-F).

[0496] The structures of these modified riboses are shown below:

[0497]

[0498] In one embodiment, at least one chemically modified nucleotide is a chemically modified phosphodiester bond.

[0499] In one embodiment, the chemically modified phosphodiester bond is selected from phosphorothioate (PS), boranophosphate, dithiophosphorothioate (PS2), 3',5'-amide, N3'-phosphoramidate (NP), phosphodiester (PO) or 2',5'-phosphodiester (2',5'-PO). In one embodiment, the chemically modified phosphodiester bond is phosphorothioate.

[0500] The structures of these modified phosphodiester bonds are shown below:

[0501]

[0502] In some embodiments, the mRNA may include a heterologous 5' untranslated region (5'UTR). In some embodiments, the mRNA may include a heterologous 3' untranslated region (3'UTR).

[0503] Methods

[0504] The compositions of the present invention can be used as compositions for diagnosing, preventing and treating diseases or disorders (e.g., infections, cancers, pain, depression, inflammatory diseases, intestinal diseases, brain diseases, allergic diseases, arrhythmias, hypertension, eye diseases, endocrine diseases or cardiovascular diseases), depending on the type of the active agent.

[0505] This document describes methods for delivering an active agent (e.g., a polynucleotide) into an organ, tissue, and / or cell, the methods comprising introducing a composition or nanoparticle described herein into the organ, tissue, and / or cell, the composition or nanoparticle comprising: a compound of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof; and an active agent.

[0506] This document describes methods for diagnosing, treating, or preventing a disease, the methods comprising administering to a subject in need thereof an effective amount of a composition or nanoparticle described herein, the composition or nanoparticle comprising a compound of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof; and an active agent.

[0507] This document describes methods for treating or preventing an infection, the methods comprising administering to a subject in need thereof an effective amount of a composition or nanoparticle described herein, the composition or nanoparticle comprising a compound of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof; and an active agent.

[0508] This document describes methods for treating or preventing a respiratory infection, the methods comprising administering to a subject in need thereof an effective amount of a composition or nanoparticle described herein, the composition or nanoparticle comprising a compound of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof; and an active agent.

[0509] This document also describes methods for inducing an immune response against a virus, the methods comprising administering to a subject in need thereof an effective amount of a composition or nanoparticle described herein, the composition or nanoparticle comprising a compound of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof; and an active agent.

[0510] This document also describes methods for inducing an immune response against a respiratory virus, the methods comprising administering to a subject in need thereof an effective amount of a composition or nanoparticle comprising a compound of formulae Da, Db, Db, Y, B, C, I-IX or any combination thereof; and an active agent.

[0511] In some embodiments, this document provides methods for delivering a polynucleotide. In some embodiments, this document provides methods for delivering a polynucleotide (e.g., mRNA) to provide expression (and translation to produce a protein) of the polynucleotide in a cell. In some embodiments, this document provides methods for delivering a polynucleotide (e.g., mRNA) to induce an immune response in a subject.

[0512] In some embodiments, a method for delivering an active agent can introduce a polynucleotide into a cell or tissue for in vivo expression of a protein encoded by the polynucleotide. In some embodiments, delivery of the active agent (e.g., polynucleotide) can correct a defect caused by a defect in the polynucleotide in the cell or tissue.

[0513] In some embodiments, a method for delivering an active agent can introduce a polynucleotide into a cell or tissue to regulate (e.g., turn off or reduce / increase) specific gene expression. For example, delivery of an antisense mRNA or shRNA to a tumor cell can regulate the expression of a protein encoded by the mRNA or shRNA.

[0514] In some embodiments, a method for delivering an active agent can introduce a polynucleotide into a cell or tissue for cell reprogramming, where the polynucleotide can be used to regulate cell behavior by expressing transcription factors or growth factors.

[0515] In some embodiments, a method for delivering an active agent can introduce a polynucleotide into in vivo cells or tissues, such as introducing a "suicide gene" or drug-sensitivity gene into tumor cells, allowing the cells to express a prodrug-activating enzyme, such as herpes simplex virus thymidine kinase, which can kill tumor cells upon contact with a prodrug (e.g., acyclovir, ganciclovir, valganciclovir, or famciclovir).

[0516] In some embodiments, a method for delivering an active agent can introduce a polynucleotide into a cell or tissue to encode a polypeptide that elicits a specific immune response against a target cell. In some embodiments, the target cell can be a cancer cell, an infected cell, or any combination thereof. In some embodiments, the polynucleotide can encode an infectious antigen polypeptide, a tumor antigen, a cytokine (e.g., interferon, interleukin, colony-stimulating factor), a monoclonal antibody, or an antibody fragment (single-chain variable fragment (scFv), fragment antigen binding ((Fab')2), intrabodies, nanobodies), or any combination thereof.

[0517] In some aspects, methods for treating cancer are disclosed herein, the methods comprising administering to a subject in need thereof an effective amount of a composition or nanoparticle described herein, the composition or nanoparticle comprising a compound of formulae Da, Db, Db, Y, B, C, I-IX, or any combination thereof; and an active agent.

[0518] In some embodiments, a method for delivering an active agent can include introducing a polynucleotide ex vivo into a specific cell or tissue to express a polypeptide encoded by the polynucleotide on the specific cell or tissue, thereby producing a chimeric cell or tissue; and administering the chimeric cell or tissue to a subject for tumor treatment. In some embodiments, the specific cell can include CAR T cells, CAR-NK cells, or other cells derived from the subject that can be used for immunotherapy.

[0519] In some embodiments, the subject is a mammal. In some embodiments, the mammal is a human. In some embodiments, the subject is a veterinary patient.

[0520] In some embodiments, the compositions herein are used to treat both local and metastatic tumors.

[0521] In some embodiments, the compositions and methods described herein can be used to treat or prevent metastasis or recurrence of cancer. In some embodiments, the compositions and methods described herein can be used to prevent recurrence of a resected solid tumor. In some embodiments, the compositions and methods described herein can be used to prevent metastasis of a resected solid tumor.

[0522] On the one hand, the methods described herein are used for treating cancer, such as melanoma, lung cancer (including adenocarcinoma of the lung, basal cell carcinoma, squamous cell carcinoma, large cell carcinoma, bronchioloalveolar carcinoma, bronchial carcinoma, non-small cell carcinoma, small cell carcinoma, mesothelioma); breast cancer (including ductal carcinoma, lobular carcinoma, inflammatory breast cancer, clear cell carcinoma, mucinous carcinoma, serous carcinoma of the serous cavity); colorectal cancer (colon cancer, rectal cancer, colorectal adenocarcinoma); anal cancer; pancreatic cancer (including pancreatic adenocarcinoma, islet cell carcinoma, neuroendocrine tumor); prostate cancer; prostatic adenocarcinoma; ovarian cancer (ovarian epithelial carcinoma or surface epithelial stromal tumor, including serous tumor, endometrioid tumor, and mucinous cystadenocarcinoma, sex cord stromal tumor); liver and bile duct cancer (including hepatocellular carcinoma, cholangiocarcinoma, hemangioma); esophageal cancer (including esophageal adenocarcinoma and squamous cell carcinoma); oral and oropharyngeal squamous cell carcinoma; adenoid cystic carcinoma of the salivary gland; bladder cancer; bladder carcinoma; uterine cancer (including endometrial adenocarcinoma, eye cancer, papillary serous carcinoma of the uterus, clear cell carcinoma of the uterus, uterine sarcoma, leiomyosarcoma, Mullerian mixed tumor); glioma, glioblastoma, medulloblastoma, and other brain tumors; kidney cancer (including renal cell carcinoma, clear cell carcinoma, Wilm's tumor); head and neck cancer (including squamous cell carcinoma); gastric cancer (gastric antrum cancer, gastric adenocarcinoma, gastrointestinal stromal tumor); testicular cancer; germ cell tumor; neuroendocrine tumor; cervical cancer; carcinoid tumors of the gastrointestinal tract, breast, and other organs; signet ring cell carcinoma; stromal tumor, including sarcoma, fibrosarcoma, cervical hemangioma, angiomatosis, hemangiopericytoma, pseudoangiomatous stromal hyperplasia, myofibroblastoma, fibromatosis, inflammatory myofibroblastoma, lipoma, angiolipoma, granular cell tumor, neurofibroma, schwannoma, angiosarcoma, liposarcoma, rhabdomyosarcoma, osteosarcoma, leiomyoma, leiomyosarcoma, skin cancer, including melanoma, cervical cancer, retinoblastoma, head and neck cancer, pancreatic cancer, brain cancer, thyroid cancer, testicular cancer, kidney cancer, bladder cancer, soft tissue cancer, adrenal cancer, urethral cancer, penile cancer, myxosarcoma, chondrosarcoma, osteosarcoma, chordoma, malignant fibrous histiocytoma, lymphangiosarcoma, mesothelioma, squamous cell carcinoma; epidermoid carcinoma, malignant skin appendage tumor, adenocarcinoma, liver cancer, hepatocellular carcinoma, renal cell carcinoma, adrenocortical tumor, hepatocellular carcinoma of the hepatobiliary duct type, transitional cell carcinoma, choriocarcinoma, seminoma, embryonal cell carcinoma, anaplastic glioma; glioblastoma multiforme, neuroblastoma, medulloblastoma, malignant meningioma, malignant schwannoma, neurofibrosarcoma, parathyroid carcinoma, medullary carcinoma of the thyroid, bronchial carcinoid, pheochromocytoma, islet cell carcinoma, malignant carcinoid, malignant paraganglioma, melanoma, Merkel cell neoplasm, cystosarcoma phyllodes, salivary cancer, thymic cancer, and vaginal cancer, etc.

[0523] In some embodiments, the compositions and methods described herein can be used to treat or prevent cancer. In some cases, the cancer is circulating cancer cells (circulating tumor cells). In some cases, the cancer is metastatic cancer cells.

[0524] In some embodiments, the method further comprises administering an additional therapeutic agent. In some embodiments, the additional therapeutic agent comprises an additional immunotherapeutic agent or an anti-tumor agent.

[0525] Also disclosed herein are methods of treating a disease or condition such as an inflammatory disorder (including an autoimmune disease) or a lymphoproliferative disorder, the method comprising administering to a subject in need thereof an effective amount of a compound, combination of compounds, or composition provided herein, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition provided herein.

[0526] Further disclosed herein are methods of treating a disease or condition such as an inflammatory disorder (including an autoimmune disease) or a lymphoproliferative disorder, the method comprising administering to a subject in need thereof an effective amount of a compound, combination of compounds, or composition provided herein, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition provided herein.

[0527] In one embodiment, provided herein is a method of treating an inflammatory disorder (including an autoimmune disease) in a subject. The method comprises administering to the subject a therapeutically effective amount of a compound, combination of compounds, or composition provided herein, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition provided herein. Examples of autoimmune diseases include, but are not limited to, acute disseminated encephalomyelitis (ADEM), Addison's disease, antiphospholipid antibody syndrome (APS), aplastic anemia, autoimmune hepatitis, autoimmune skin diseases, celiac disease, Crohn's disease, diabetes (type 1), Goodpasture's syndrome, Graves' disease, Guillain-Barré syndrome (GBS), Hashimoto's disease, lupus erythematosus, multiple sclerosis, myasthenia gravis, opsoclonus myoclonus syndrome (OMS), optic neuritis, Ord's thyroiditis, pemphigus, polyarthritis, primary biliary cirrhosis, psoriasis, rheumatoid arthritis, Reiter's syndrome, Takayasu's arteritis, temporal arteritis (also known as "giant cell arteritis"), warm autoimmune hemolytic anemia, Wegener's granulomatosis, alopecia totalis (e.g., inflammatory hair loss), Chagas disease, chronic fatigue syndrome, akinetic mutism, endometriosis, hidradenitis suppurativa, interstitial cystitis, neuromyotonia, sarcoidosis, scleroderma, ulcerative colitis, vitiligo, and vulvodynia. Other diseases include bone resorption disorders and thrombosis.

[0528] Inflammation has multiple forms and includes, but is not limited to, acute, adhesive, atrophic, catarrhal, chronic, cirrhotic, diffuse, disseminated, exudative, fibrinous, fibrotic, focal, granulomatous, hyperplastic, hypertrophic, interstitial, metastatic, necrotic, obliterative, parenchymal, plastic, productive, proliferative, pseudomembranous, purulent, sclerotic, serofibrinous, serous, simple, specific, subacute, suppurative, toxic, traumatic, and / or ulcerative inflammation.

[0529] Exemplary inflammatory conditions include, but are not limited to, inflammation associated with acne, anemia (e.g., aplastic anemia, hemolytic autoimmune anemia), asthma, arteritis (e.g., polyarteritis, temporal arteritis, periarteritis nodosa, Takayasu's arteritis), arthritis (e.g., crystalline arthritis, osteoarthritis, psoriatic arthritis, gout attack, gouty arthritis, reactive arthritis, rheumatoid arthritis, and Reiter's arthritis), ankylosing spondylitis, amyloidosis, amyotrophic lateral sclerosis, autoimmune diseases, allergies or allergic reactions, atherosclerosis, bronchitis, bursitis, chronic prostatitis, conjunctivitis, Chagas disease, chronic obstructive pulmonary disease, dermatomyositis, diverticulitis, diabetes (e.g., type I diabetes, type 2 diabetes), skin diseases (e.g., psoriasis, eczema, burns, dermatitis, pruritus (itching)), endometriosis, Guillain - Barré syndrome, infections, ischemic heart disease, Kawasaki disease, glomerulonephritis, gingivitis, hypersensitivity, headache (e.g., migraine, tension headache), intestinal obstruction (e.g., postoperative intestinal obstruction and intestinal obstruction during sepsis), idiopathic thrombocytopenic purpura, interstitial cystitis (bladder pain syndrome), gastrointestinal disorders (e.g., selected from peptic ulcer, Crohn's disease, diverticulitis, gastrointestinal bleeding, eosinophilic gastrointestinal disorders (e.g., eosinophilic esophagitis, eosinophilic gastritis, eosinophilic gastroenteritis, eosinophilic colitis), gastritis, diarrhea, gastroesophageal reflux disease (GORD or its synonym GERD), inflammatory bowel disease (IBD) (e.g., Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, diversion colitis, Behcet's syndrome, indeterminate colitis) and irritable bowel syndrome (IBS)), lupus, multiple sclerosis, morphea, myasthenia gravis, myocardial ischemia, nephrotic syndrome, pemphigus vulgaris, pernicious anemia, peptic ulcer, polymyositis, primary biliary cirrhosis, neuroinflammation associated with brain disorders (e.g., Parkinson's disease, Huntington's disease, and Alzheimer's disease), prostatitis, chronic inflammation associated with cranial radiation injury, pelvic inflammatory disease, polymyalgia rheumatica, reperfusion injury, Crohn's disease, rheumatic fever, systemic lupus erythematosus, scleroderma, sclerosis, sarcoidosis, spondyloarthropathy, Sjogren's syndrome, thyroiditis, transplant rejection, tendinitis, trauma or injury (e.g., frostbite, chemical irritants, toxins, scars, burns, physical injury), vasculitis, vitiligo, and Wegener's granulomatosis.In certain embodiments, the inflammatory disorder is selected from arthritis (e.g., rheumatoid arthritis), inflammatory bowel disease, inflammatory bowel syndrome, asthma, psoriasis, endometriosis, interstitial cystitis, and prostatitis. In certain embodiments, the inflammatory condition is an acute inflammatory condition (e.g., inflammation caused by an infection). In certain embodiments, the inflammatory condition is a chronic inflammatory condition (e.g., conditions caused by asthma, arthritis, and inflammatory bowel disease). The compound can also be used to treat inflammation associated with trauma and non-inflammatory myalgia.

[0530] Immune disorders such as autoimmune disorders include, but are not limited to, arthritis (including rheumatoid arthritis, spondyloarthropathy, gouty arthritis, degenerative joint diseases such as osteoarthritis, systemic lupus erythematosus, Sjogren's syndrome, ankylosing spondylitis, undifferentiated spondylitis, Behcet's disease, hemolytic autoimmune anemia, multiple sclerosis, amyotrophic lateral sclerosis, amyloidosis, acute shoulder pain, psoriasis, and juvenile arthritis), asthma, atherosclerosis, osteoporosis, bronchitis, tendinitis, bursitis, skin diseases (e.g., psoriasis, eczema, burns, dermatitis, pruritus (itching)), enuresis, eosinophilic diseases, gastrointestinal disorders (e.g., selected from peptic ulcer, regional enteritis, diverticulitis, gastrointestinal bleeding, eosinophilic gastrointestinal diseases (e.g., eosinophilic esophagitis, eosinophilic gastritis, eosinophilic gastroenteritis, eosinophilic colitis), gastritis, diarrhea, gastroesophageal reflux disease (GORD or its synonym GERD), inflammatory bowel disease (IBD) (e.g., Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, diversion colitis, Behcet's syndrome, indeterminate colitis), and inflammatory bowel syndrome (IBS)), relapsing polychondritis (e.g., atrophic polychondritis and systemic polychondromalacia), and conditions improved by the following: prokinetics (e.g., ileus during intestinal obstruction, postoperative ileus, and sepsis; gastroesophageal reflux disease (GORD or its synonym GERD); eosinophilic esophagitis, gastroparesis such as diabetic gastroparesis; food intolerance and food allergy, and other functional bowel disorders such as non-ulcer dyspepsia (NUD) and non-cardiac chest pain (NCCP, including costochondritis)).

[0531] In one embodiment, provided herein is a method of treating an infection in a subject, the infection being caused by an infectious agent. The method comprises administering to the subject a therapeutically effective amount of a compound, combination of compounds, or composition provided herein, or a pharmaceutically acceptable form thereof, or a pharmaceutical composition provided herein. Infectious agents can include, but are not limited to, bacterial organisms such as Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyogenes, Corynebacterium diphtheriae, Listeria monocytogenes, Bacillus anthracis, Clostridium tetani, Clostridium botulinum, Clostridium perfringens, Neisseria meningitidis, Neisseria gonorrhoeae, Streptococcus mutans, Pseudomonas aeruginosa, Salmonella typhi, Haemophilus parainfluenzae, Bordetella pertussis, Francisella tularensis, Yersinia pestis, Vibrio cholerae, Legionella pneumophila, Mycobacterium tuberculosis, Mycobacterium leprae, Treponema pallidum, Leptospira interrogans, Borrelia burgdorferi, Campylobacter jejuni, etc.; viruses such as metapneumovirus such as human metapneumovirus (Hmpv), parainfluenza virus such as human parainfluenza virus (Hpiv) types 1, 2, and 3 (Hpiv1, Hpiv2, and Hpiv3, respectively), respiratory syncytial virus (RSV), measles virus (MeV), coronaviruses (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HcoV-OC43, HcoV-229E, HcoV-NL63, HcoV-NL, HcoV-NH, HcoV-HKU1), poxviruses (e.g., smallpox, monkeypox), African swine fever virus, influenza A and B, human immunodeficiency virus (HIV), varicella-zoster, herpes simplex 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, Coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A, B, C, D, and E viruses, etc.; fungi, protozoa, and parasitic organisms such as Cryptococcus neoformans, Histoplasma capsulatum, Candida albicans, Candida tropicalis, Nocardia asteroides, Rickettsia rickettsii, Rickettsia typhi, Mycoplasma pneumoniae, Chlamydia psittaci, Chlamydia trachomatis, Plasmodium falciparum, Trypanosoma brucei, Entamoeba histolytica, Toxoplasma gondii, Trichomonas vaginalis, Schistosoma mansoni, etc.

[0532] In some embodiments, the infection can be a coronavirus infection. In one embodiment, the coronavirus infection is an infection of the upper respiratory tract and / or lower respiratory tract. "Upper respiratory tract" includes the mouth, nose, sinuses, middle ear, throat, larynx, and trachea. "Lower respiratory tract" includes the bronchi (bronchial tubes) and lungs (bronchi, bronchioles, and alveoli) and the interstitial tissue of the lungs.

[0533] In another embodiment, the coronavirus infection is a gastrointestinal infection. "Gastrointestinal tract" can include any region of the tube from the mouth to the anus, including the mouth, esophagus, stomach, and intestines.

[0534] In yet another embodiment, the coronavirus infection is a kidney infection.

[0535] It should be understood and is contemplated herein that the coronavirus infections disclosed herein can give rise to a pathological condition associated with the coronavirus infection, referred to herein as "coronavirus disease". In some embodiments, the coronavirus disease is selected from the common cold, pneumonia, pulmonary inflammation, bronchitis, severe acute respiratory syndrome (SARS), coronavirus disease 2019 (COVID-2019), Middle East respiratory syndrome (MERS), sinusitis, swine diarrhea, porcine epidemic diarrhea, avian infectious bronchitis, otitis, and pharyngitis. In some embodiments, the coronavirus infection is the common cold. In some embodiments, the coronavirus infection is selected from SARS, COVID-19, and MERS. In a particular embodiment, the coronavirus infection is COVID-19. In another particular embodiment, the coronavirus infection is IBV, PorCoV HKU15, or PEDV.

[0536] Most SARS patients were previously healthy adults between the ages of 25 and 70. Some suspected SARS cases have been reported in children under 15 years of age. The case fatality rate for patients meeting the current World Health Organization definitions of probable and suspected SARS cases was approximately 3%.

[0537] Other indications associated with coronavirus infections are described in Gralinski and Baric, 2015, Journal of Pathology 235:185-195 and Cavanagh, 2005, "Coronaviridae: a review of coronavirus and toroviruses", Coronaviruses with Special Emphasis on First Insights Concerning SARS 1, edited by A. Schmidt, M. H. Wolff, and O. Weber, Birkhauser Verlag Baser, Switzerland, each of which is incorporated herein by reference in its entirety.

[0538] The coronavirus causing the infection can be selected from alpha coronavirus, beta coronavirus, gamma coronavirus, or delta coronavirus.

[0539] Ocular disorders that can be treated according to the compositions and methods disclosed herein include Acanthamoeba keratitis, fungal keratitis, bacterial keratitis, viral keratitis, onchocercal keratitis, bacterial keratoconjunctivitis, viral keratoconjunctivitis, corneal dystrophic diseases, Fuchs' endothelial dystrophy, Sjogren's syndrome, Stevens-Johnson syndrome, autoimmune dry eye, environmental dry eye, corneal neovascularization diseases, prevention and treatment of rejection after corneal transplantation, autoimmune uveitis, infectious uveitis, anterior uveitis, posterior uveitis (including toxoplasmosis), panuveitis, vitreous or retinal inflammatory diseases, prevention and treatment of endophthalmitis, macular edema, macular degeneration, age-related macular degeneration, proliferative and non-proliferative diabetic retinopathy, hypertensive retinopathy, retinal autoimmune diseases, primary and metastatic intraocular melanoma, other intraocular metastatic tumors, open-angle glaucoma, closed-angle glaucoma, pigmentary glaucoma, and combinations thereof.

[0540] Endocrine disorders that can be treated according to the compositions and methods disclosed herein include thyroid disorders (e.g., hyperthyroidism, hypothyroidism, thyroiditis, goiter), diabetes, hypoglycemia, glucagonoma, calcium homeostasis disorders (e.g., parathyroid, osteoporosis, osteomalacia, rickets), sex hormone disorders (e.g., sexual development disorders, hypogonadism, puberty disorders, menstrual function or fertility disorders).

[0541] 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 disclosed invention belongs. Publications and materials cited herein are specifically incorporated by reference.

[0542] All compositions and methods disclosed and claimed herein can be prepared and implemented according to the present disclosure without undue experimentation. While the compositions and methods of the present disclosure have been described in connection with preferred embodiments, it will be apparent to those skilled in the art that changes can be made to the compositions and methods and to the steps or the order of steps in the methods described herein, and such changes do not depart from the spirit, scope, and concept of the present disclosure. More specifically, it will be apparent that certain chemically related agents can replace the agents described herein while achieving the same or similar results. It is apparent to those skilled in the art that all such similar substitutions and modifications are considered to be within the spirit, scope, and concept of the present disclosure as defined by the appended claims.

[0543] By way of non-limiting illustration, examples of certain embodiments of the present disclosure are given below.

[0544] Example

[0545] Example 1:

[0546] Figure 1 Describes the synthetic route of compound 4 (iVe).

[0547] General procedure for preparing compound 1.

[0548]

[0549] 7-Bromoheptanoic acid (2.98 g, 11.6 mmol) and heptadecane-9-ol (1.5 g, 5.8 mmol) were dissolved in dichloromethane (20 mL), and then N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (1.1 g, 5.8 mmol), N,N-diisopropylethylamine (3.3 mL, 18.7 mmol) and DMAP (0.114 g, 0.9 mmol) were added to the solution. The reaction was stirred at room temperature for 18 hours. The reaction was diluted with dichloromethane and extracted with saturated NaHCO3 (aqueous solution). The organic layer was separated, washed with brine, then dried over NaSO4, filtered and evaporated in vacuo. The residue was purified by silica gel chromatography (0-10% ethyl acetate / hexane) to afford compound 1 (907 mg, 42%). HR-MS (ESI) m / z = 447.2830.

[0550] General procedure for preparing compound 2.

[0551]

[0552] 6-Bromohexanoyl chloride (0.93 mL, 6 mmol, 1.2 equiv) was added dropwise to a stirred solution of vitamin E (5 mmol, 2.15 g, 1.0 equiv) and NEt3 (1.1 mL, 7.5 mmol, 1.5 equiv) in anhydrous CH2Cl2 (25 mL) at 0 °C. The mixture was warmed to room temperature and stirred for more than 6 hours. After the reaction was completed, the solvent was removed in vacuo and the crude product was purified by column chromatography (0-20% EA / PE) to give compound 2 in high yield (>80%). HR-MS (ESI) m / z = 607.3712.

[0553] General procedure for preparing compound 3.

[0554]

[0555] A solution of compound 2 (3.8 g, 8.2 mmol) and 4-amino-1-butanol (22.1 g, 248 mmol) in ethanol (3 mL) was stirred at 62 °C for 18 h. The reaction mixture was concentrated in vacuo and the residue was extracted with ethyl acetate and water. The organic layer was separated, washed with water and brine, dried over Na2SO4, filtered and evaporated in vacuo. The residue was purified by silica gel chromatography [0 - 100% (a mixture of 0.5% NH4OH, 20% MeOH in dichloromethane) / dichloromethane] to afford compound 3 (yield: 41%). HR-MS (ESI) m / z = 616.5440.

[0556] General procedure for the preparation of compound 4 (iVe).

[0557] A solution of compound 1 (4.5 g, 10 mmol), compound 3 (3.1 g, 5 mmol), KI (0.93 g, 5.5 mmol) and K2CO3 (2.76 g, 20 mmol) in a dry acetonitrile / dichloromethane / cyclopropyl methyl ether (3:2:1, 120 mL) mixture was stirred at 80 °C for 18 h. The reaction was cooled to room temperature, filtered and the solvent was evaporated in vacuo. The residue was purified by silica gel chromatography (0 - 10% MeOH / dichloromethane) to afford compound 4 (iVe) (1.1 g, 22%). HR-MS (ESI) m / z = 982.8554.

[0558] 1 1H NMR spectrum ( Figure 2A ), 13 13C NMR spectrum ( Figure 2B ) and mass spectrum ( Figure 2C ) confirmed the structure. Figure 3 The size distribution of representative lipid nanoparticles (LNPs) formulated with iVe lipid and eGPF-Luc mRNA is shown. Table 1 summarizes the physicochemical properties of LNPs formulated with iVe lipid and eGPF-Luc mRNA.

[0559] Table 1.

[0560] Diameter (nm) 92.2 Polydispersity index (PDI) 0.02 ζ - potential (mV) -5.0 Encapsulation efficiency (EE) 93.2％

[0561] The in vitro expression of GFP and luciferase was tested in 293T cells transfected with 1.0 μg eGFP-Luc mRNA at 1.0 μg / mL mRNA for 24 h. Figure 4A Representative fluorescence images of 293T cells after transfection are shown. Figure 4B The transfection efficacy analysis of GFP expression using flow cytometry is shown. Figure 4CThe transfection effect of luciferase expression using bioluminescence analysis was shown.

[0562] The in vivo transfection efficiency of mRNA expressing luciferase was tested. The LNP was formulated with iVe lipid, and 1 μg of the formulated mRNA expressing luciferase was intramuscularly injected. After administration, representative images of luciferase expression measured by whole-body bioluminescence imaging of dLN using the IVIS Spectrum in vivo imaging system at 4, 24, and 48 hours are shown in Figure 5A and the injection site is shown in Figure 5B And (5C - 5D). Quantitative luciferase expression measured by whole-body bioluminescence imaging of dLN using the IVIS Spectrum in vivo imaging system at 4, 24, and 48 hours is shown in Figure 5C and the injection site is shown in Figure 5D And.

[0563] Example 2:

[0564] Figure 6 The synthetic route of iVd was described.

[0565] General procedure for preparing Compound 2.

[0566]

[0567] Compound 1 (30.0 g, 131 mmol, 1.00 equivalent) was charged into R1 (1.00 L three-necked flask) at 20 °C. DCM (350 mL) was charged into R1 at 20 °C. Compound 1a (27.4 g, 131 mmol, 1.00 equivalent) was charged into R1 at 20 °C. DIEA (67.9 g, 525 mmol, 91.5 mL, 4.00 equivalents), DMAP (3.21 g, 26.2 mmol, 0.20 equivalent), EDCI (32.7 g, 170 mmol, 1.30 equivalents) were charged into R1 at 20 °C. The mixture was stirred at 20 °C for 16 h. TLC (petroleum ether:ethyl acetate = 20:1) showed that most of Compound 1 (R f = 0.60) was consumed and a new main spot (R f = 0.50) was detected. H2O (300 mL) was charged into R2 (1.00 L bucket) at 20 °C. The reaction was charged into R2 at 20 °C. The reaction was extracted with dichloromethane (300 mL * 3) at 20 °C. The reaction was washed with saturated brine (200 mL * 2) at 20 °C. The reaction was dried over anhydrous Na2SO4 at 20 °C. The organic phase was concentrated below 40 - 50 °C to obtain a residue. The residue was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 50:1 to 10:1, R fPurification was carried out with (= 0.50). Compound 2 as a colorless liquid (30.0 g, 71.5 mmol, 54.4% yield) was obtained.

[0568] 1 H NMR: 400 MHz, CDCl3 δ 4.89 - 4.86 (m, 1H), 3.42 - 3.39 (m, 2H), 2.30 (t, J=7.6 Hz, 2H), 1.87 (m, 2H), 1.70 - 1.65 (m, 2H), 1.58

[0569] General procedure for the preparation of Compound 3.

[0570]

[0571] Compound 2 (30.0 g, 71.5 mmol, 1.00 equiv) was charged into R1 (250 mL three - necked bottom flask) at 20 °C. EtOH (30.0 mL) was charged into R1 at 20 °C. Compound 2a (191 g, 2.15 mol, 199 mL, 30.0 equiv) was charged into R1 at 20 °C. The mixture was heated to 65 °C. The mixture was stirred at 65 °C for 16 h. TLC (dichloromethane:methanol = 5:1) showed that most of Compound 2 (R f = 0.95) was consumed and a new main spot (R f = 0.20) was detected. H2O (100 mL) was charged into R2 (250 mL bucket) at 20 °C. The reaction was charged into R2 at 20 °C. The reaction was extracted with ethyl acetate (60.0 mL * 3) at 20 °C. The reaction was washed with saturated brine (50.0 mL * 3) at 20 °C. The reaction was dried over anhydrous Na2SO4 at 20 °C. The organic phase was concentrated below 40 - 50 °C to obtain a residue. The residue was purified by column chromatography (SiO2, dichloromethane:methanol = 50:1 to 10:1, R f = 0.20). Compound 3 as a yellow oil (17.0 g, 39.7 mmol, 55.5% yield) was obtained.

[0572] 1 H NMR: 400 MHz, CDCl3 δ 4.88 - 4.82 (m, 1H), 3.58 - 3.55 (m, 2H), 2.66 - 2.59 (m, 4H), 2.27 - 2.19 (m, 2H), 1.64 - 1.53 (m, 6H), 1.44 - 1.42 (m, 6H), 1.28 - 1.25 (m, 7H), 1.19 (s, 17H), 0.89 - 0.85 (m, 6H).

[0573] General procedure for the preparation of Compound 4 - 2.

[0574]

[0575] Charge compound 4-1 (5.00 g, 13.0 mmol, 1.10 eq) into R1 (250 mL three-necked flask) at 20 °C. Charge DCM (100 mL) into R1 at 20 °C. Charge compound 3-2 (2.31 g, 11.8 mmol, 1.00 eq) into R1 at 20 °C. Charge EDCI (2.95 g, 15.3 mmol, 1.30 eq), DIEA (6.00 g, 47.2 mmol, 8.23 mL, 4.00 eq), DMAP (288 mg, 2.36 mmol, 0.20 eq) into R1 at 20 °C. Stir the mixture at 20 °C for 24 h. TLC (petroleum ether:ethyl acetate = 5:1) showed that most of compound 4-1 (R f = 0.20) was consumed and a new main spot (R f = 0.80) was detected. Charge H2O (100 mL) into R2 (250 mL bucket) at 20 °C. Charge the reaction into R2 at 20 °C. Extract the reaction with dichloromethane (50 mL * 3) at 20 °C. Dry the reaction with anhydrous Na2SO4 at 20 °C. Concentrate the organic phase below 40 - 50 °C to obtain a residue. Purify the residue by column chromatography (SiO2, petroleum ether:ethyl acetate = 5:1 to 1:1, R f = 0.70). Obtain compound 4-2 (4.20 g, 7.48 mmol, 63.2% yield) as a yellow oil.

[0576] 1 1H NMR: 400 MHz, CDCl3 δ 6.21 (d, J = 11.2 Hz, 1H), 6.03 (d, J = 11.2 Hz, 1H), 5.06–4.84 (m, 3H), 3.42–3.38 (m, 2H), 2.83–2.80 (m, 1H), 2.59 - 2.56 (m, 1H), 2.55–2.39 (m, 4H), 2.33–2.29 (m, 1H), 1.89–1.87 (m, 6H), 1.66–1.64 (m, 1H), 1.51 - 1.49 (m, 4H), 1.48–1.47 (m, 6H), 1.46–1.45 (m, 4H), 1.44–1.42 (m, 2H), 1.37–1.35 (m, 3H), 1.34–1.33 (m, 1H), 1.32–1.31 (m, 3H), 0.93–0.86 (m, 6H), 0.55 (s, 3H).

[0577] General procedure for the preparation of iVd.

[0578]

[0579] Compound 4-2 (1.50 g, 2.67 mmol, 1.00 equiv) was charged into R1 (500 mL three-necked flask) at 20 °C. ACN (30.0 mL) was charged into R1 at 20 °C. Compound 3 (1.14 g, 2.67 mmol, 1.00 equiv) was charged into R1 at 20 °C. K2CO3 (1.48 g, 10.6 mmol, 4 equiv), KI (487 mg, 2.94 mmol, 1.10 equiv), CMPE (6.97 g, 69.5 mmol, 8.10 mL, 26.0 equiv) were charged into R1 at 20 °C. The mixture was heated to 90 °C. The mixture was stirred at 90 °C for 16 h. TLC (dichloromethane:methanol = 5:1) showed that most of the reactant 1 (R f = 0.20) was consumed and a new main spot (R f = 0.50) was detected. H2O (80.0 mL) was charged into R2 (250 mL bucket) at 20 °C. The reaction was charged into R2 at 20 °C. The reaction was extracted with dichloromethane (80.0 mL * 3) at 20 °C. The reaction was dried with anhydrous Na2SO4 at 20 °C. The organic phase was concentrated below 40 - 50 °C to obtain a residue. The residue was purified by column chromatography (SiO2, dichloromethane:methanol = 100:1 to 10:1, R f = 0.50). The target 4 (iVd) was obtained as a colorless oil (0.50 g, 1.09 mmol, 40.8% yield).

[0580] MS: Product: m / z = 909.1 (M + H) +

[0581] HPLC: Product: R t = 3.541 min, 100% purity at 220 nm.

[0582] 1 1H NMR: 400 MHz, CDCl3 δ 6.21 (d, J = 11.2 Hz, 1H), 6.03 (d, J = 11.2 Hz, 1H), 5.08–4.84 (m, 4H), 2.29 (s, 2H), 2.83–2.80 (m, 1H), 2.58–2.55 (m, 18H), 2.48 - 2.37 (m, 5H), 2.29–2.26 (m, 15H), 1.66–1.63 (m, 13H), 1.61–1.51 (m, 26H), 1.36–1.34 (m, 3H), 1.32 - 1.30 (m, 1H), 1.26–1.13 (m, 12H), 0.55 (s, 3H).

[0583] 1 The \(^1\)H NMR spectrum ( Figure 7A ), mass spectrometry ( Figure 7B ), and HPLC ( Figure 7C ) confirmed the structure. Table 2 shows the size distribution, PDI, ζ-potential, and encapsulation efficiency of representative lipid nanoparticles (LNPs) formulated with iVd lipid and eGFP-Luc mRNA.

[0584] Table 2.

[0585] Diameter (nm) 112.8 Polydispersity index (PDI) 0.09 ζ - potential (mV) -4.0 Encapsulation efficiency (EE) 91.2％

[0586] The in vitro expression of GFP was tested in 293T cells transfected with 1.0 μg of eGFP mRNA at 1.0 μg / mL mRNA for 24 hours. Figures 8B - 8D Representative fluorescence images of 293T cells after transfection are shown. Figure 8A Transfection efficacy analysis of GFP expression using flow cytometry is shown.

[0587] The in vivo transfection efficiency of luciferase-expressing mRNA was tested. LNPs were formulated with iVd lipid and 1 μg of the formulated luciferase-expressing mRNA was intramuscularly injected. Representative images of luciferase expression measured by whole-body bioluminescence imaging of dLN and injection site using the IVIS Spectrum in vivo imaging system at 4, 24, 48, and 96 hours after administration are shown in Figures 9A - 9D . Quantitative luciferase expression measured by whole-body bioluminescence imaging of dLN, injection site, and liver using the IVIS Spectrum in vivo imaging system at 4, 24, 48, and 96 hours is shown in Figures 9E - 9G .

[0588] The compositions and methods of the appended claims are not limited to the scope of the specific compositions and methods described herein. The methods are intended to be illustrative of several aspects of the claims, and any compositions and methods that are functionally equivalent are intended to fall within the scope of the claims. Various modifications to the methods, in addition to the compositions and methods shown and described herein, are intended to fall within the scope of the appended claims. Further, although only certain representative compositions and method steps are specifically described herein, other combinations of these compositions and method steps are also intended to fall within the scope of the appended claims, even if not specifically recited. Thus, combinations of steps, elements, components, or ingredients may be explicitly recited herein, however, other combinations including steps, elements, components, and ingredients, even if not explicitly stated.

Claims

1. A compound of formula Da, Db or Dc: Wherein: Z is a vitamin; X is O or S; L 1 is -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-; L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -NR a C(=O)- or -C(=O)R a N-; L 3 Absent, being -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-; R a where n is an integer from 0 to 12; x is 0, 1 or 2; M is N, CH, D is N or G 1 、G 2 、G 3 and G 5 each independently selected from C1-C 12 alkyl or C1-C 12 alkenyl; G 4 absent or selected from C1-C 12 alkyl or C1-C 12 alkenyl; R 2 is -CR b R c , C6-C 24 alkyl or C6-C 24 alkenyl; R 3 For R 6 is H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , R b and R c are independently H, C1-C 12 alkyl or C1-C 12 alkenyl; and p, q and r are independently 0, 1, 2, 3 or 4; or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, wherein the vitamin moiety can be selected from: Wherein R 1 , R 4 and R 10 are each independently hydrogen or C1-C6 alkyl; and Represents a carbon-carbon bond or a carbon-carbon double bond.

3. A compound of formula Y, formula B or formula C: Wherein: R 1 、R 4 and R 10 each independently represents hydrogen or a C1-C6 alkyl group; represents a carbon-carbon bond or a carbon-carbon double bond; A is X is O or S; L 1 is -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-; L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -NR a C(=O)- or -C(=O)R a N-; L 3 Absent, being -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-; R a where n is an integer from 0 to 12; x is 0, 1 or 2; M is N, CH, D is N or G 1 、G 2 、G 3 and G 5 each independently selected from C1-C 12 alkyl or C1-C 12 alkenyl; G 4 is absent or selected from C1-C 12 alkyl or C1-C 12 alkenyl; R 2 is -CR b R c , C6-C 24 alkyl or C6-C 24 alkenyl; R 3 For R 6 is H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , R b and R c are independently H, C1-C 12 alkyl or C1-C 12 alkenyl; and p, q and r are independently 0, 1, 2, 3 or 4; or a pharmaceutically acceptable salt thereof.

4. The compound according to any one of claims 1 to 3, wherein the compound of formula Y, B or C can be selected from: or a pharmaceutically acceptable salt thereof or any combination thereof; Where A is the same as formula Y, formula B or formula C.

5. A compound of formula I, II, III, IV, V, VI, VII, VIII or IX: Wherein: R 1 、R 4 and R 10 each independently represents hydrogen or a C1-C6 alkyl group; represents a carbon-carbon single bond or a carbon-carbon double bond; X is O or S; L 1 is -O(C=O)-, -C(=O)-, -S(O) x -, -C(=O)R a C(=O)-, -NR a C(=O)-, -NC(=O)R a C(=O)- or -C(=O)NR a C(=O)-; L 2 is -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -S(O) x -, -S-S-, -NR a C(=O)- or -C(=O)R a N-; L 3 Absent, being -O(C=O)-, -(C=O)O-, -O(C=O)O-, -C(=O)-, -O-, -NR a C(=O)- or -C(=O)R a N-; R a where n is an integer from 0 to 12; x is 0, 1 or 2; M is N, CH, D is N or G 1 、G 2 、G 3 and G 5 each independently selected from C1-C 12 alkyl or C1-C 12 alkenyl; G 4 is absent or selected from C1-C 12 alkyl or C1-C 12 alkenyl; R 2 is -CR b R c , C6-C 24 alkyl or C6-C 24 alkenyl; R 3 For R 6 is H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , R b and R c are independently H, C1-C 12 alkyl or C1-C 12 alkenyl; and p, q and r are independently 0, 1, 2, 3 or 4; or a pharmaceutically acceptable salt thereof.

6. The compound according to any one of claims 1 to 5, wherein X is O.

7. The compound according to any one of claims 2 to 6, wherein R 4 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl or hexyl.

8. The compound according to any one of claims 2 to 6, wherein R 10 is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl or hexyl.

9. The compound according to any one of claims 2 to 7, wherein R 10 is hydrogen 10. The compound according to any one of claims 1 to 9, wherein M is CH.

11. The compound according to any one of claims 1 to 10, wherein R 3 is 12. The compound according to any one of claims 1 to 9, wherein R 6 is H, OR b , CN, -C(=O)OR b , -NC(=O)R b , -C(=O)NR b , 13. A compound according to any one of claims 1 to 12, wherein L 1 is -C(=O)-.

14. The compound according to any one of claims 1 to 13, wherein L 2 is -(C=O)O-.

15. The compound according to any one of claims 1 to 14, wherein L 3 is absent.

16. The compound according to any one of claims 1 to 15, wherein G 1 is a C5 alkyl group.

17. A compound according to any one of claims 1 to 16, wherein G 2 is a C6 alkyl group.

18. A compound according to any one of claims 1 to 17, wherein G 3 is a C2 alkyl group.

19. A compound according to any one of claims 1 to 18, wherein G 4 is absent.

20. The compound according to any one of claims 1 to 19, wherein R 2 is -CR b R c .

21. The compound according to any one of claims 1 to 10, wherein R 3 is 22. The compound according to any one of claims 1 to 10, wherein R 3 is 23. The compound according to any one of claims 1 to 21, wherein R b and R c is a C8 alkyl group.

24. The compound according to any one of claims 1 to 22, wherein p, r and q are 0.

25. The compound according to any one of claims 1 to 9, 11 or 13 to 23, wherein M is N.

26. The compound according to any one of claims 1 to 25, wherein the compounds of formula Y, B, C, I-XII can be selected from: or a pharmaceutically acceptable salt thereof.

27. A composition comprising: The compound according to any one of claims 1 to 26; and An active agent.

28. The composition according to claim 27, wherein the active agent is a diagnostic agent, a prophylactic agent or a therapeutic agent.

29. The composition according to any one of claims 27 to 28, wherein the active agent is a nucleic acid, a protein / peptide, a small molecule or any combination thereof.

30. The composition according to 29, wherein the agent is a nucleic acid.

31. The composition according to any one of claims 29 to 30, wherein the nucleic acid comprises double-stranded DNA, single-stranded DNA, composite DNA, encapsulated DNA, naked RNA, encapsulated RNA, messenger RNA (mRNA), tRNA, short interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), antisense RNA (asRNA), self-amplifying mRNA (saRNA), guide RNA (gRNA), cRNA or any combination thereof.

32. The composition according to any one of claims 29 to 31, wherein the nucleic acid comprises mRNA.

33. The composition according to claim 32, wherein the mRNA encodes at least one metapneumovirus (hMPV) antigen polypeptide, parainfluenza virus (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3 respectively) antigen polypeptides, respiratory syncytial virus (RSV) antigen polypeptide, measles virus (MeV) antigen polypeptide, varicella-zoster virus antigen polypeptide, influenza virus antigen polypeptide, herpes simplex virus 1 (HSV1) antigen polypeptide, herpes simplex virus 2 (HSV2) antigen polypeptide, poxvirus (e.g., smallpox, monkeypox) antigen polypeptide, African swine fever virus antigen polypeptide, cytomegalovirus antigen polypeptide, Epstein-Barr virus antigen polypeptide, rotavirus antigen polypeptide, rhinovirus antigen polypeptide, adenovirus antigen polypeptide, papillomavirus antigen polypeptide, poliovirus antigen polypeptide, mumps antigen polypeptide, rabies antigen polypeptide, rubella antigen polypeptide, coxsackievirus antigen polypeptide, equine encephalitis antigen polypeptide, Japanese encephalitis antigen polypeptide, yellow fever antigen polypeptide, Rift Valley fever antigen polypeptide, hepatitis A, B, C, D, and E virus antigen polypeptides, or coronavirus (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, HCoV-HKU1) antigen polypeptide, or any combination thereof.

34. The composition according to claim 33, wherein the coronavirus is a human coronavirus.

35. The composition according to claim 34, wherein the mRNA encodes a tumor antigen, a cytokine (e.g., interferon, interleukin, colony-stimulating factor), a monoclonal antibody, or an antibody fragment (single-chain variable fragment (scFv), fragment antigen binding ((Fab')2), intracellular antibody, nanobody), or any combination thereof.

36. The composition according to any one of claims 27 to 35, wherein the composition comprises a lipid nanoparticle dispersion, a liposome formulation, a lipid emulsion, or any combination thereof.

37. A lipid nanoparticle comprising: Greater than 0% to 50% of the compound according to any one of claims 1 to 26; 0% to 50% of a cationic lipid; 0% to 80% of an ionizable lipid; 0% to 5% of a polyethylene glycolylated lipid; 0% to 40% of a co-lipid; 0% to 80% of a sterol; And An active agent encapsulated in the nanoparticle.

38. The nanoparticle according to claim 37, wherein the active agent comprises a nucleic acid, a protein / peptide, a small molecule, or any combination thereof.

39. The nanoparticle according to claim 38, wherein the active agent is a nucleic acid.

40. The nanoparticle according to any one of claims 38 to 39, wherein the nucleic acid comprises double-stranded DNA, single-stranded DNA, complex DNA, encapsulated DNA, naked RNA, encapsulated RNA, messenger RNA (mRNA), tRNA, short interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), antisense RNA (asRNA), self-amplifying mRNA (saRNA), guide RNA (gRNA), cRNA, or any combination thereof.

41. The nanoparticle according to any one of claims 38 to 40, wherein the nucleic acid comprises mRNA.

42. The nanoparticle according to claim 41, wherein the mRNA encodes a tumor antigen, a cytokine (e.g., interferon, interleukin, colony-stimulating factor), a monoclonal antibody, or an antibody fragment (single-chain fragment variable (scFv), fragment antigen binding ((Fab')2), intracellular antibody, nanobody), or any combination thereof.

43. The nanoparticle according to claim 42, wherein the mRNA encodes at least one human metapneumovirus (hMPV) antigen polypeptide, parainfluenza virus (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3, respectively) antigen polypeptides, respiratory syncytial virus (RSV) antigen polypeptides, measles virus (MeV) antigen polypeptides, varicella-zoster virus antigen polypeptides, influenza virus antigen polypeptides, herpes simplex virus 1 (HSV1) antigen polypeptides, herpes simplex virus 2 (HSV2) antigen polypeptides, poxvirus (e.g., smallpox, monkeypox) antigen polypeptides, African swine fever virus antigen polypeptides, cytomegalovirus antigen polypeptides, Epstein-Barr virus antigen polypeptides, rotavirus antigen polypeptides, rhinovirus antigen polypeptides, adenovirus antigen polypeptides, papillomavirus antigen polypeptides, poliovirus antigen polypeptides, mumps antigen polypeptides, rabies antigen polypeptides, rubella antigen polypeptides, coxsackievirus antigen polypeptides, equine encephalitis antigen polypeptides, Japanese encephalitis antigen polypeptides, yellow fever antigen polypeptides, Rift Valley fever antigen polypeptides, hepatitis A, B, C, D, and E virus antigen polypeptides, or coronavirus (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, HCoV-HKU1) antigen polypeptides, or any combination thereof.

44. The nanoparticle according to claim 43, wherein the coronavirus is a human coronavirus.

45. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and an effective amount of the lipid nanoparticle according to any one of claims 37 to 44.

46. A method for delivering an active agent into a cell, the method comprising: introducing the composition according to any one of claims 27 to 36 or the nanoparticle according to any one of claims 37 to 44 into the cell.

47. A method for treating or preventing an infection caused by an infectious agent, the method comprising administering to a subject in need thereof a composition according to any one of claims 27 to 36, a nanoparticle according to any one of claims 37 to 44.

48. A method for inducing an immune response against an infection caused by an infectious agent, the method comprising administering to a subject in need thereof a composition according to any one of claims 27 to 36, a nanoparticle according to any one of claims 37 to 44.

49. The method according to any one of claims 47 to 48, wherein the infective agent comprises bacterial organisms such as Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyrogenes, Corynebacterium diphtheriae, Listeria monocytogenes, Bacillus anthracis, Clostridium tetani, Clostridium botulinum, Clostridium perfringens, Neisseria meningitidis, Neisseria gonorrhoeae, Streptococcus mutans, Pseudomonas aeruginosa, Salmonella typhi, Haemophilus parainfluenzae, Bordetella pertussis, Francisella tularensis, Yersinia pestis, Vibrio cholerae, Legionella pneumophila, Mycobacterium tuberculosis, Mycobacterium leprae, Treponema pallidum, Leptospira interrogans, Borrelia burgdorferi, Campylobacter jejuni, etc.;Viruses, such as metapneumoviruses such as human metapneumovirus (hMPV), parainfluenza viruses such as human parainfluenza virus (hPIV) types 1, 2, and 3 (hPIV1, hPIV2, and hPIV3, respectively), respiratory syncytial virus (RSV), measles virus (MeV), coronaviruses (e.g., MERS-CoV, SARS-CoV, SARS-CoV2, HCoV-OC43, HCoV-229E, HCoV-NL63, HCoV-NL, HCoV-NH, HCoV-HKU1), poxviruses (e.g., smallpox, monkeypox), African swine fever virus, influenza A and B, human immunodeficiency virus (HIV), varicella-zoster, herpes simplex 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, Coxsackievirus, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A, B, C, D, and E viruses, etc.; fungi, protozoa, and parasites, such as Cryptococcus neoformans, Histoplasma capsulatum, Candida albicans, Candida tropicalis, Nocardia asteroides, Rickettsia rickettsii, Rickettsia typhi, Mycoplasma pneumoniae, Chlamydia psittaci, Chlamydia trachomatis, Plasmodium falciparum, Trypanosoma brucei, Entamoeba histolytica, Toxoplasma gondii, Trichomonas vaginalis, Schistosoma mansoni, etc.

50. A method for treating or preventing a respiratory infection, the method comprising administering to a subject in need thereof a composition according to any one of claims 27 to 36, a nanoparticle according to any one of claims 37 to 44.

51. A method for inducing an immune response against a respiratory virus, the method comprising administering to a subject in need thereof a composition according to any one of claims 27 to 36, a nanoparticle according to any one of claims 37 to 44.

52. The method according to claim 51, wherein the respiratory infection is caused by metapneumovirus (MPV), human parainfluenza virus (hPIV) types 1, 2 and 3 (hPIV1, hPIV2 and hPIV3 respectively), RSV, measles virus (MeV) or coronavirus infection.

53. The method according to claim 52, wherein the coronavirus is a human coronavirus.

54. The method according to any one of claims 47 to 53, wherein the subject is a veterinary patient.

55. The method according to any one of claims 47 to 53, wherein the subject is a mammal.

56. The method according to claim 55, wherein the mammal is a human.

57. The method according to any one of claims 46 to 56, which can be administered orally, topically, transdermally, transepithelially, intra-articularly, intra-arterially, intradermally, intraventricularly, intralesionally, intranasally, rectally, vaginally, by inhalation, by an implanted reservoir, subcutaneously, intravenously, intramuscularly, intra-articularly, intrasynovially, intrasternal, intrathecally, intraperitoneally, intrahepatically, intralesionally and intracranially by injection or infusion techniques.

58. A method for delivering an active agent into a cell, the method comprising: introducing a composition according to any one of claims 27 to 36 or a nanoparticle according to any one of claims 37 to 44 into the cell.

59. The method according to claim 58, wherein the composition or nanoparticle comprises an active agent, the active agent comprising a nucleic acid, a protein / peptide, a small molecule or any combination thereof.

60. The method according to claim 56, wherein the active agent comprises a nucleic acid.

61. The method according to any one of claims 58 to 60, which comprises introducing a composition or nanoparticle comprising a nucleic acid into a specific cell or tissue to express a protein encoded by the nucleic acid in order to correct a defect caused by the lack of the nucleic acid in the cell or tissue.

62. The method according to any one of claims 58 to 60, comprising in vivo introducing a composition or nanoparticle comprising a nucleic acid into a specific cell or tissue to express a protein encoded by the nucleic acid on the specific cell or tissue so as to induce an immune response in the subject.

63. The method according to any one of claims 58 to 60, comprising ex vivo introducing a composition or nanoparticle comprising a nucleic acid into a specific cell or tissue to express a protein encoded by the nucleic acid on the specific cell or tissue, thereby generating a chimeric cell or tissue; and administering the chimeric cell or tissue to a subject.

64. The method according to any one of claims 58 to 60, wherein the specific cell is a T cell (e.g., CAR T cell) derived from the subject.

65. The method according to any one of claims 58 to 60, wherein the method introduces a polynucleotide to regulate gene expression, protein expression, or any combination thereof.

66. The method according to any one of claims 58 to 60, wherein the method introduces a polynucleotide to reprogram cells by regulating cell behavior.

67. The method according to any one of claims 58 to 60, wherein the polynucleotide encodes an intracellular antibody.

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