Trehalose-based surfactant

Abstract

Compounds that function as surfactants and / or excipients, methods for producing such compounds, methods for using such compounds, and compositions comprising such compounds, preferably in combination with one or more additional agents or therapies, are presented herein.

Description

Technical Field

[0001] Compounds that function as surfactants and / or excipients, methods for producing such compounds, methods for using such compounds, and compositions comprising such compounds, preferably in combination with one or more additional agents or therapies, are provided herein.

Background Art

[0002] Excipients are a large class of molecules used in pharmaceutical formulations of chemical pharmaceuticals, antibodies, and proteins. They are also involved in improving the storage life stability of biomolecules during storage, or in the cryopreservation and lyophilization of biomolecules.

[0003] Excipients used in pharmaceutical formulations of antibodies and proteins can be classified as follows. · Bulking agents, such as the polyol / disaccharide / polysaccharide family and natural polysaccharides and amino acids, · Isotonic agents based on sugar materials, · Buffering agents such as organic and inorganic salts, · Surfactants for reducing surface tension while improving the solubility of drugs, · Antioxidants, · Chelating agents, · Preservatives

[0004] Excipients are important for the biomaterial preservation process in the research and development industry. Biomolecules are usually cryopreserved at -80 °C or lyophilized. Maintaining cryopreserved materials is a huge cost in terms of electricity and nitrogen. Also, lyophilization is not very expensive but is accompanied by significant loss of active substances. It is difficult to keep the material intact when the material composition (loss of water), temperature, and pressure are impaired. Polymers are subject to chemical instability under stress conditions (temperature changes, exposure to light, oxygen, or chemical stress and shear stress), which may result in loss of biological activity. This instability can refer to irreversible denaturation (where a protein loses its tertiary or secondary structure) and aggregation (where a protein self-organizes irreversibly).

[0005] Among surfactants, polysorbates PS20 and PS80 are widely approved for the parenteral administration of antibodies. However, they have been reported to impair long-term storage due to the presence of peroxide residues from the poly(ethylene glycol) moiety. This causes protein oxidation and increases proteolysis. Furthermore, polysorbates have been shown to damage cell membrane integrity and mitochondrial function at a concentration of 1 - 2 v / v% on BEAS-2B bronchial epithelial cells.

Summary of the Invention

Problems to be Solved by the Invention

[0006] Therefore, better and safer excipients must be developed for the stability of polymers, for long-term storage, and, if necessary, for the administration of polymers.

Means for Solving the Problems

[0007] The present invention relates to a compound of formula I

Chemical Formula

[0008] The present invention also provides a composition comprising a compound of formula I described herein.

[0009] The present invention further provides a pharmaceutical composition comprising a compound of formula I described herein.

[0010] The present invention further provides a method for preparing a compound of formula I, comprising: · sequentially and selectively functionalizing two primary alcohols of trehalose while maintaining other hydroxyl moieties unreacted; · conjugating first to a fatty acid chain and then to a carboxylated side chain, wherein the carboxyl functional group is introduced in the form of a benzyl ester, which benzyl ester is removed in the last step of the procedure. The present invention further provides a method for preparing a polymer of a compound of formula I, comprising self - condensing an activated trehalose derivative in the presence of a coupling agent.

[0011] BRIEF DESCRIPTION OF THE DRAWINGS Aggregation monitored over time by DLS during storage at 4 °C. The graph shows the total area under the curve (AUC) of the peaks corresponding to antibody aggregates for 1 mg / mL of antibody in different stabilization media after 0, 5, and 4 days of storage.

[0012]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Mode for Carrying Out the Invention

[0013] As used in the present disclosure, the following words and phrases are generally intended to have the meanings described below, unless otherwise explicitly indicated or unless they are clearly inconsistent with the context in which they are used.

[0014] The following description shows exemplary methods, parameters, etc. However, it should be recognized that such description is not intended as a limitation on the scope of the present disclosure, but is provided as an explanation of exemplary embodiments.

[0015] As used herein, the following words, phrases and symbols are generally intended to have the meanings described below, unless they are clearly inconsistent with the context in which they are used.

[0016] A dash ("-") that is not between two letters or symbols is used to indicate the point of attachment of a substituent. For example, -CH 2 is bonded via a carbon atom. Dashes at the beginning or end of a chemical group are for convenience. Chemical groups may be depicted with or without one or more dashes without losing their normal meaning. Unless chemically or structurally required, the order in which chemical groups are described or named does not indicate directionality or imply it.

[0017] The prefix "C u-v " indicates that the following group has u to v carbon atoms. For example, "C 1-6"Alkyl" indicates that the alkyl group has 1 to 6 carbon atoms.

[0018] References to "about" a value or parameter in this specification include (and describe) embodiments that are directed to the value or parameter itself. In certain embodiments, the term "about" includes the recited amount ±10%. In other embodiments, the term "about" includes the recited amount ±5%. In certain other embodiments, the term "about" includes the recited amount ±1%. Also, the term "about X" includes the recitation of "X". Also, the singular forms "a" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, a reference to "a compound" includes a plurality of such compounds, and a reference to "an assay" includes a reference to one or more assays known to those of skill in the art and their equivalents.

[0019] The term "substituted" means that any one or more hydrogen atoms on the specified atom or group are replaced by one or more substituents other than hydrogen, provided that the normal valence of the specified atom is not exceeded. Examples of one or more substituents include, but are not limited to, alkyl, alkenyl, alkynyl, alkoxy, acyl, amino, amide, amidino, aryl, azide, carbamoyl, carboxyl, carboxyl ester, cyano, guanidino, halo, haloalkyl, heteroalkyl, heteroaryl, heterocycloalkyl, hydroxy, hydrazino, imino, oxo, nitro, alkylsulfinyl, sulfonic acid, alkylsulfonyl, thiocyanate, thiol, thione, or combinations thereof. Similar indeterminate structures (e.g., a substituted aryl having a substituted alkyl, where the substituted alkyl itself is substituted with a substituted aryl group, and the substituted aryl group is further substituted with a substituted heteroalkyl group, etc.) arrived at by defining substituents using polymers or further substituents added ad infinitum are not intended to be included herein. Unless otherwise specified, the maximum number of consecutive substitutions in the compounds described herein is 3. For example, consecutive substitution of a substituted aryl group by two other substituted aryl groups is limited to ((substituted aryl)substituted aryl)substituted aryl. Similarly, the above definitions are not intended to include unacceptable substitution patterns (e.g., a methyl substituted with 5 fluorines or a heteroaryl group having 2 adjacent oxygen ring atoms). Such unacceptable substitution patterns are well known to those skilled in the art. When used to modify a chemical group, the term "substituted" may describe other chemical groups as defined herein. For example, the term "substituted aryl" includes, but is not limited to, "alkylaryl". Unless otherwise noted, when a group is described as being optionally substituted, any substituent of that group is itself unsubstituted.

[0020] The "substituted" group also includes embodiments in which a monovalent substituent is attached to one atom of the substituted group (e.g., forms a branch), and embodiments in which the substituent may be a divalent bridging group that forms a condensed ring on the substituted group by attaching to two adjacent atoms of the substituted group.

[0021] "Alkyl" refers to an unbranched or branched saturated hydrocarbon chain. As used herein, alkyl has from 1 to 20 carbon atoms (i.e., C 1-20 alkyl), from 1 to 8 carbon atoms (i.e., C 1-8 alkyl), from 1 to 6 carbon atoms (i.e., C 1-6 alkyl), or from 1 to 4 carbon atoms (i.e., C 1-4 alkyl). Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and 3-methylpentyl. When an alkyl residue having a specific number of carbon atoms is named by chemical name or specified by a molecular formula, all positional isomers having that number of carbon atoms may be included. Thus, for example, "butyl" includes n-butyl (i.e., -(CH 2 ) 3 CH 3 ), sec-butyl (i.e., -CH(CH 3 )CH 2 CH 3 ), isobutyl (i.e., -CH 2 CH(CH 3 ) 2 ) and tert-butyl (i.e., -C(CH 3 ) 3 ), and "propyl" includes n-propyl (i.e., -(CH 2 ) 2 CH 3 ) and isopropyl (i.e., -CH(CH 3 ) 2 ).

[0022] "Alkenyl" refers to an aliphatic group containing at least one carbon-carbon double bond and having 2 to 20 carbon atoms (i.e., C 2-20 alkenyl), having 2 to 8 carbon atoms (i.e., C 2-8 alkenyl), having 2 to 6 carbon atoms (i.e., C 2-6 alkenyl), or having 2 to 4 carbon atoms (i.e., C 2-4 alkenyl). Examples of alkenyl groups include ethenyl, propenyl, and butadienyl (including 1,2-butadienyl and 1,3-butadienyl).

[0023] "Alkynyl" refers to an aliphatic group containing at least one carbon-carbon triple bond and having 2 to 20 carbon atoms (i.e., C 2-20 alkynyl), having 2 to 8 carbon atoms (i.e., C 2-8 alkynyl), having 2 to 6 carbon atoms (i.e., C 2-6 alkynyl), or having 2 to 4 carbon atoms (i.e., C 2-4 alkynyl). The term "alkynyl" also includes groups having one triple bond and one double bond.

[0024] "Alkoxy" refers to the group "alkyl - O -" or "-O - alkyl". Examples of alkoxy groups include methoxy, ethoxy, n - propoxy, isopropoxy, n - butoxy, tert - butoxy, sec - butoxy, n - pentoxy, n - hexoxy, and 1,2 - dimethylbutoxy.

[0025] "Amino" refers to the group NR y R z , where in the formula, R y and R z are independently selected from hydrogen, alkyl, haloalkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl, each of which may optionally be substituted.

[0026] The term "alkylsulfinyl" refers to the group -SO-alkyl, where alkyl is as defined above and includes optionally substituted alkyl groups as also defined above.

[0027] "Cycloalkyl" refers to a saturated or partially saturated cyclic alkyl group having a monocyclic or polycyclic ring system including fused ring systems, bridged ring systems and spiro ring systems. As used herein, cycloalkyl has 3 to 20 ring carbon atoms (i.e., C 3-20 cycloalkyl), 3 to 12 ring carbon atoms (i.e., C 3-12 cycloalkyl), 3 to 10 ring carbon atoms (i.e., C 3-10 cycloalkyl), 3 to 8 ring carbon atoms (i.e., C 3-8 cycloalkyl), or 3 to 6 ring carbon atoms (i.e., C 3-6 cycloalkyl). Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0028] As used herein, the term "cycloalkenyl" group means a non-aromatic carbocyclic group having at least one double bond.

[0029] "Cyanoalkyl" refers to an alkyl group substituted with cyano (CN).

[0030] "Halogen" or "halo" includes fluoro, chloro, bromo and iodo.

[0031] The term "haloalkyl" refers to a monovalent or divalent group having the indicated carbon atoms of an alkyl group in which one or more hydrogen atoms are substituted with halogen. Examples of haloalkyl groups include -CH 2 F, -CHF 2 , -CF 3 , -CH 2 CF 3 , -CHFCH 2 F, -CF 2 -, -CHF-, etc. Similarly, the term "haloalkoxy", for example -O-C1-3 Haloalkyl refers to an alkoxy group in which one or more hydrogen atoms of an alkyl group are replaced by halogen. Examples of haloalkoxy groups include -OCH 2 F, -OCHF 2 , -OCF 3 , -OCH 2 CF 3 , -OCHFCH 2 F, etc. Those skilled in the art will recognize that similar definitions apply to the above alkenyl and alkynyl analogs (e.g., C 2-4 haloalkenyl, -O-C 2-4 haloalkynyl).

[0032] "Heteroalkyl" refers to an alkyl group in which one or more carbon atoms (and any accompanying hydrogen atoms) are each independently replaced by the same or different heteroatom groups. The term "heteroalkyl" includes unbranched or branched saturated chains having carbon and heteroatoms. As an example, one, two, or three carbon atoms may each independently be replaced by the same or different heteroatom groups. Examples of heteroatom groups include -NR-, -O-, -S-, -SO-, -SO 2 -, etc., but are not limited thereto, where R is H, alkyl, aryl, cycloalkyl, heteroalkyl, heteroaryl, or heterocycloalkyl, each of which may optionally be substituted. Examples of heteroalkyl groups include -OCH 3 , -CH 2 OCH 3 , -SCH 3 , -CH 2 SCH 3 , -NRCH 3 , and -CH 2 NRCH 3 , where R is hydrogen, alkyl, aryl, arylalkyl, heteroalkyl, or heteroaryl, each of which may optionally be substituted. As used herein, heteroalkyl contains 1 to 10 carbon atoms, 1 to 8 carbon atoms, or 1 to 4 carbon atoms, and 1 to 3 heteroatoms, 1 to 2 heteroatoms, or 1 heteroatom.

[0033] "Heterocycloalkyl" refers to a saturated or unsaturated cyclic alkyl group having one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur. Heterocycloalkyl may be monocyclic or polycyclic, and this polycycle may be fused, bridged, or spiro. As used herein, heterocycloalkyl has 1 to 5 ring heteroatoms independently selected from nitrogen, sulfur, or oxygen, 1 to 4 ring heteroatoms independently selected from nitrogen, sulfur, or oxygen, 1 to 3 ring heteroatoms independently selected from nitrogen, sulfur, or oxygen, 1 to 2 ring heteroatoms independently selected from nitrogen, sulfur, or oxygen, or 1 ring heteroatom selected from nitrogen, sulfur, or oxygen, and has 2 to 20 ring carbon atoms (i.e., C 2-20 heterocycloalkyl), has 2 to 12 ring carbon atoms (i.e., C 2-12 heterocycloalkyl), has 2 to 10 ring carbon atoms (i.e., C 2-10 heterocycloalkyl), has 2 to 8 ring carbon atoms (i.e., C 2-8 heterocycloalkyl), has 3 to 12 ring carbon atoms (i.e., C 3-12 heterocycloalkyl), has 3 to 8 ring carbon atoms (i.e., C 3-8 heterocycloalkyl), or has 3 to 6 ring carbon atoms (i.e., C 3-6(heterocycloalkyl). Examples of heterocycloalkyl groups include pyrrolidinyl, piperidinyl, piperazinyl, oxetanyl, dioxolanyl, azetidinyl, and morpholinyl. As used herein, the term "bridged heterocycloalkyl" refers to a 4- to 10-membered cyclic moiety linked at two non-adjacent atoms of the heterocycloalkyl, wherein one or more (e.g., one or two) 4- to 10-membered cyclic moieties have at least one heteroatom, and each heteroatom is independently selected from nitrogen, oxygen, and sulfur. As used herein, bridged heterocycloalkyl includes bicyclic and tricyclic ring systems. Also as used herein, the term "spiro-heterocycloalkyl" refers to a ring system in which a 3- to 10-membered heterocycloalkyl has one or more additional rings, and the one or more additional rings are 3- to 10-membered cycloalkyl or 3- to 10-membered heterocycloalkyl, and one atom of the one or more additional rings is also an atom of the above 3- to 10-membered heterocycloalkyl. Examples of spiro-heterocycloalkyl include bicyclic and tricyclic ring systems such as 2-oxa-7-azaspiro[3.5]nonanyl, 2-oxa-6-azaspiro[3.4]octanyl, and 6-oxa-1-azaspiro[3.3]heptanyl.

[0034] "Acyl" refers to the group -C(=O)R, where R is hydrogen, alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroalkyl, or heteroaryl, each of which may be optionally substituted as defined herein. Examples of acyl include formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethyl-carbonyl, and benzoyl.

[0035] The term "N-alkylated" means that one of the hydrogen atoms of a mono-substituted amine, or a di-substituted amine group or a tri-substituted amine group is substituted by an alkyl group. When alkylation is carried out on a tri-substituted amine group, an alkonium salt is formed, i.e., a positive charge is generated on the nitrogen atom. N-alkylation is usually related to alkyl substitution on a ring nitrogen atom.

[0036] The term "oxo" refers to the group =O.

[0037] The term "carboxy" refers to the group -C(O)-OH.

[0038] The term "ester" or "carboxyl ester" refers to the group -C(O)OR, where R is alkyl, cycloalkyl, aryl, heteroaryl, or heterocyclyl, which may optionally be further substituted, for example, by alkyl, alkoxy, halogen, CF 3 , amino, substituted amino, cyano or -SO n R f (wherein R f is alkyl, aryl or heteroaryl and n is 0, 1 or 2) may be further substituted.

[0039] The term "substituted amino" refers to the group -NRR, where each R is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, or heterocyclyl (each of which may optionally be substituted), or a group described or exemplified herein, or both R groups are linked to form a heterocyclic group (e.g., morpholino) described or exemplified herein, which may also optionally be substituted.

[0040] The term "amide" refers to the group -C(O)NRR, wherein each R is independently hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, or heterocyclyl (each of which may optionally be substituted), or a group described or exemplified herein, or both R groups are linked to form a heterocyclic group (e.g., morpholino) described or exemplified herein, which may also optionally be substituted.

[0041] The term "sulfoxide" refers to the group -SOR, wherein R is alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl, each of which may optionally be substituted.

[0042] As used herein, the terms "alkylcycloalkyl", "alkylaryl", "alkylheteroaryl" and "alkylheterocyclyl" are intended to refer to a cycloalkyl, aryl, heteroaryl or heterocyclyl group attached to the remainder of the molecule through an alkyl moiety, and the terms "alkyl", "cycloalkyl", "aryl", "heteroaryl" and "heterocyclyl" are as defined herein. Exemplary alkylaryl groups include benzyl, phenethyl and the like.

[0043] "Optional" or "optionally" means that the event or circumstance described below may or may not occur, and that the description includes both the case where the event or circumstance occurs and the case where it does not occur.

[0044] As used herein, polyethylene glycol or "PEG" refers to a polymer represented as H-(O-CH2-CH2)n-OH, where n represents the number of times the O-CH2-CH2 (oxyethylene) moiety is repeated, and n may vary widely since PEG results in a wide variety of molecular weights. For example, n may be about 33 for low molecular weight polyethylene glycol (about 1500 g / mol), may range up to about 227 for high molecular weight polyethylene glycol (about 10,000 g / mol), may be about 454 for PEG with a molecular weight of about 20,000 g / mol, may be 908 for PEG with a molecular weight of about 40,000, and may be even higher for higher molecular weight PEG varieties.

[0045] Certain commonly used alternative chemical names may be used. For example, divalent groups such as divalent "alkyl" groups and divalent "aryl" groups may also be referred to as "alkylene" groups or "alkylenyl" groups, "arylene" groups or "arylenyl" groups, respectively. Also, unless expressly stated otherwise, when a combination of groups is referred to herein as one moiety, e.g., arylalkyl, the last-mentioned group contains the atom through which the moiety is attached to the remainder of the molecule.

[0046] A given group (moiety) is described herein as being attached to a second group, and where the site of attachment is not explicit, this given group may be attached at any available site of the given group or at any available site of the second group. For example, “alkyl-substituted phenyl” may, where the site of attachment is not explicit, have any available site of the alkyl group attached to any available site of the phenyl group. In this regard, an “available site” is a site of the group where the hydrogen of the group may optionally be replaced by a substituent.

[0047] It is understood that in all of the substituted groups defined above, polymers reached by defining a substituent having further substituents on the substituent itself (for example, a substituted aryl having a substituted aryl group as a substituent, where the substituted aryl group as this substituent is itself substituted by a substituted aryl group, etc.) are not intended to be encompassed herein. Regardless of whether the substituents are the same or different, an infinite number of substituents are also not included. In such cases, the maximum number of such substituents is 3. Thus, each of the above definitions is restricted, for example, by the limitation that a substituted aryl group is limited to -substituted aryl-(substituted aryl)-substituted aryl.

[0048] “Isomers” are different compounds having the same molecular formula. Isomers include stereoisomers, enantiomers and diastereomers.

[0049] “Stereoisomers” are isomers that differ only in the way the atoms are arranged in space.

[0050] “Enantiomers” are pairs of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of an enantiomer pair is a “racemic” mixture. The term “(±)” is used, where appropriate, to indicate a racemic mixture.

[0051] “Diastereoisomers” are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other.

[0052] The compounds of the present disclosure may have one or more chiral centers and may be produced as racemic mixtures or as individual enantiomers or diastereoisomers. The number of stereoisomers present in any given compound of a given formula depends on the number of chiral centers present (if n is the number of chiral centers, 2 n n individual stereoisomers are possible). The individual stereoisomers may be obtained by resolution of a racemic or non-racemic mixture of intermediates at some suitable stage of synthesis or by resolution of the compounds by conventional means. The individual stereoisomers (including individual enantiomers and diastereoisomers), as well as racemic and non-racemic mixtures of stereoisomers, are included within the scope of the present disclosure, and all of them are intended to be depicted by the structures herein unless specifically indicated otherwise.

[0053] Absolute stereochemistry is specified according to the Cahn Ingold Prelog RS system. When a compound is a pure enantiomer, the stereochemistry of each chiral carbon may be specified by R or S. A resolved compound of unknown absolute configuration may be designated as (+) or (-) according to the direction (dextrorotatory or levorotatory) in which the compound rotates the plane of polarization of the sodium D line wavelength.

[0054] As used herein, "trehalose" refers to a non-reducing disaccharide consisting of two glucose units. Trehalose is found in many plants, microorganisms, and animals and is involved in glucose storage, signaling and regulation, structural and transport roles, and as a protectant and stabilizer of membranes and proteins. It is widely approved in biopharmaceutical formulations for pharmaceutical development. Trehalose stabilizes biomolecules by its ability to form hydrogen bonds with amino acids present on the surface of the biomolecules.

[0055] The term "polymorph" refers to different crystal structures of a crystalline compound. Different polymorphs may arise from differences in crystal packing (packing polymorphs) or from differences in packing between different conformational isomers of the same molecule (conformational polymorphs).

[0056] The term "solvate" refers to a complex formed by combining a compound of formula (I) or any other compound of any formula disclosed herein with a solvent.

[0057] The term "hydrate" refers to a complex formed by the combination of a compound of formula (I) or any compound of any formula disclosed herein with water.

[0058] The term "polymorph", as used herein, refers to different crystalline forms of the same compound, as well as other solid molecular forms including pseudopolymorphs such as hydrates (e.g., bound water present in the crystal structure as discussed above) and solvates (e.g., bound solvents other than water) of the same compound. Different crystalline polymorphs have different crystal structures due to different packings of the molecules in the lattice. This results in different crystal symmetries and / or unit cell parameters that directly affect physical properties such as the X-ray diffraction characteristics of the crystal or powder. For example, different polymorphs generally diffract at different sets of angles and give different values for intensity. Therefore, X-ray powder diffraction can be used to identify different polymorphs, or solid forms containing multiple polymorphs, in a reproducible and reliable manner (S. Byrn et al., Pharmaceutical Solids: A Strategic Approach to Regulatory Considerations, Pharmaceutical research, Vol. 12, No. 7, pp. 945 - 954, 1995; J. K. Haleblian and W. McCrone, Pharmacetical Applications of Polymorphism, Journal of Pharmaceutical Sciences, Vol. 58, No. 8, pp. 911 - 929, 1969).

[0059] The term "prodrug" refers to a compound of formula (I) or a derivative of formula (I) disclosed herein that contains a chemical group that can be converted in vivo and / or cleaved from the remainder of the molecule to provide an active drug. Pharmaceutically acceptable salts of prodrugs of compounds of formula (I) or their biologically active metabolites are also within the scope of the present disclosure.

[0060] Any formula or structure given herein that includes formula (I), or any formula disclosed herein, is intended to represent both the unlabeled form and the isotopically labeled form of the compound. Isotopically labeled compounds have the structure depicted by the formula given herein, except that one or more atoms are replaced by an isotope having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as 2 H (deuterium, D), 3 H (tritium), 11 C, 13 C, 14 C, 15 N, 18 F, 31 P, 32 P, 35 S, 36 Cl and 125 I, among others, are included but not limited to. Various isotopically labeled compounds of the present disclosure, such as 3 H, 13 C and 14Those incorporating radioisotopes such as C are within the scope of the present disclosure. Such isotope-labeled compounds may be useful in metabolic studies, kinetics studies, detection or imaging techniques, such as positron emission tomography (PET) or single photon emission computed tomography (SPECT) including tissue distribution assays of drugs or substrates, or in the treatment of patients. Such isotope-labeled analogs of the compounds of the present disclosure may provide improved pharmacokinetic and / or pharmacodynamic properties over the unlabeled form of the same compound and thus may also be useful in the treatment of the diseases disclosed herein. Such isotopically equilibrated forms or analogs of the compounds herein are within the scope of the present disclosure. One of ordinary skill in the art can prepare and use such isotopically labeled forms according to procedures for isotopically labeling a compound or a moiety of a compound to arrive at an isotope-labeled or radiolabeled analog of a compound disclosed herein.

[0061] The term "pharmaceutically acceptable salt" of a given compound refers to salts that retain the biological effectiveness and properties of the given compound and are not undesirable from a biological or other perspective. Pharmaceutically acceptable base addition salts can be prepared from inorganic bases and organic bases. Salts derived from inorganic bases include, by way of example only, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts and magnesium salts. Salts derived from organic bases include primary, secondary and tertiary amines, such as alkylamines, dialkylamines, trialkylamines, substituted alkylamines, di(substituted alkyl)amines, tri(substituted alkyl)amines, alkenylamines, dialkenylamines, trialkenylamines, substituted alkenylamines, di(substituted alkenyl)amines, tri(substituted alkenyl)amines, cycloalkylamines, di(cycloalkyl)amines, tri(cycloalkyl)amines, substituted cycloalkylamines, disubstituted cycloalkylamines, trisubstituted cycloalkylamines, cycloalkenylamines, di(cycloalkenyl)amines, tri(cycloalkenyl)amines, substituted cycloalkenylamines, disubstituted cycloalkenylamines, trisubstituted cycloalkenylamines, arylamines, diarylamines, triarylamines, heteroarylamines, diheteroarylamines, triheteroarylamines, heterocyclic amines, diheterocyclic amines, triheterocyclic amines, and salts of mixed diamines and triamines in which at least two of the substituents on the amine are different and are selected from alkyl, substituted alkyl, alkenyl, substituted alkenyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, heteroaryl, heterocyclic, etc., but are not limited thereto. Also included are amines in which two or three substituents together with the amino nitrogen form a heterocyclic or heteroaryl group. The amine has the general structure N(R 30 )(R 31 )(R 32 ), and monosubstituted amines have two of the three substituents (R 30 , R 31 , and R 32 ) on nitrogen as hydrogen, and disubstituted amines have three substituents (R 30 , R31 and R 32 ) has one of them as hydrogen, but the trisubstituted amine has 3 two substituents (R 30 , R 31 , and R 32 ) none of which is hydrogen. R 30 , R 31 , and R 32 is selected from various substituents such as hydrogen, optionally substituted alkyl, aryl, heteroaryl, cycloalkyl, cycloalkenyl, heterocyclyl, etc.

[0062] Specific examples of suitable amines include, by way of example only, isopropylamine, trimethylamine, diethylamine, tri(isopropyl)amine, tri(n-propyl)amine, ethanolamine, diethanolamine, 2-dimethylaminoethanol, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucosamine, theobromine, purine, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like.

[0063] Pharmaceutically acceptable acid addition salts may be prepared from inorganic acids and organic acids. Salts derived from inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Salts derived from organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluene-sulfonic acid, salicylic acid, and the like.

[0064] Sugar-based surfactants with diverse structures can be good alternatives to polysorbate surfactants. They are mainly produced by enzymatic synthesis from microorganisms. They exhibit low toxicity, biodegradability, and biocompatibility and are produced from more inexpensive and renewable energy sources. Applications in pharmaceuticals, food, cosmetics, textiles, oil, and agriculture are interested in using biosurfactants (bio-surfactants) for their benefits. However, their use in the pharmaceutical industry is limited due to the risk of endotoxin (endotoxin) contamination of biosurfactants made from bacteria. Their production results in a mixture of glycolipids with different properties, leading to conflicting and confusing results. More recently, sugar-based surfactants have been synthesized. Among these, trehalose lipids are composed of trehalose linked to lipids via ester bonds at various positions.

[0065] Compounds that function as surfactants and / or excipients, methods for producing such compounds, methods for using such compounds, and compositions comprising such compounds, preferably in combination with one or more additional agents or therapies, are provided herein.

[0066] All aspects regarding the compound are further contemplated to include any pharmaceutically acceptable salts, stereoisomers, polymorphs, mixtures of stereoisomers, solvates or prodrugs thereof.

[0067] In some aspects, the present disclosure provides a compound of formula (I)

Chemical formula

[0068] In one embodiment, R 1 is -OH. In one embodiment, R 2 is attached to R 3 through oxygen. In one embodiment, R 4 is OH. In one embodiment, X 1 is -(C=O)-CH2-CH2-(C=O). In one embodiment, Z 1 is -OH. In one embodiment, X 2 is -(C=O). In one embodiment, Z 2 is alkyl, alkenyl, or absent. Examples of alkyl include, as described above, alkyl having 1 to 20 carbon atoms (i.e., C1-20 alkyl), alkyl having 1 to 8 carbon atoms (i.e., C1-8 alkyl), in one embodiment, alkyl having 1 to 6 carbon atoms (i.e., C1-6 alkyl), or alkyl having 1 to 4 carbon atoms (i.e., C1-4 alkyl), but are not limited thereto. Examples of alkenyl include alkenyl having 2 to 20 carbon atoms (i.e., C2-20 alkenyl), alkenyl having 2 to 8 carbon atoms (i.e., C2-8 alkenyl), alkenyl having 2 to 6 carbon atoms (i.e., C2-6 alkenyl), or alkenyl having 2 to 4 carbon atoms (i.e., C2-4 alkenyl), but are not limited thereto. Examples of alkenyl groups include ethenyl, propenyl, butadienyl (including 1,2-butadienyl and 1,3-butadienyl).

[0069] In one embodiment, the compound of formula (I) is selected from the group consisting of the following.

Table 1(1)

Table 1(2)

[0070] The present disclosure provides, in some embodiments, a composition comprising a compound of formula (I) of the present invention. In some preferred embodiments, the compound of formula (I) is a surfactant and / or an excipient and / or exhibits the properties of a surfactant and / or an excipient.

[0071] The present disclosure further provides, in some embodiments, a pharmaceutical composition comprising a compound of formula (I) of the present invention. This pharmaceutical composition further comprises a therapeutically effective amount of a pharmaceutical agent.

[0072] In some embodiments, the pharmaceutical agent is selected from the group consisting of chemical compounds, peptides, lipids, oligonucleotides, cell exosomes, and combinations of one or more thereof.

[0073] Examples of peptides include peptides selected from the group consisting of antibodies, antigen-binding fragments, and combinations thereof.

[0074] As used herein, "antibody" or "antigen-binding protein or polypeptide" refers to a polypeptide or polypeptide complex that specifically recognizes and binds to an antigen. An antibody may be a whole antibody and any antigen-binding fragment or its single chain. Thus, the term "antibody" includes any protein or peptide-containing molecule that includes at least a portion of an immunoglobulin molecule having the biological activity of binding to an antigen. Examples of such include, but are not limited to, the complementarity-determining regions (CDRs) of the heavy or light chain or the ligand-binding portion thereof, the variable regions of the heavy or light chain, the constant regions of the heavy or light chain, the framework (FR) regions, or any portion thereof, or at least one portion of a binding protein.

[0075] The term "antibody fragment" or "antigen-binding fragment" as used herein refers to F(ab’) 2 , F(ab) 2 , Fab’, Fab, Fv, scFv, and the like, which are parts of an antibody. Regardless of structure, an antibody fragment binds to the same antigen as the intact antibody recognizes. The term "antibody fragment" includes aptamers, spiegelmers, and diabodies (bispecific antibodies). The term "antibody fragment" also includes any synthetic or genetically engineered protein that acts like an antibody by binding to a specific antigen to form a complex.

[0076] The antibody may be derived from any animal origin, including birds and mammals. Preferably, the antibody is a human antibody, a mouse antibody, a donkey antibody, a rabbit antibody, a goat antibody, a guinea pig antibody, a camel antibody, a llama antibody, a horse antibody, or a chicken antibody. In another aspect, the antibody or antigen-binding protein is a humanized antibody or a humanized antigen-binding protein.

[0077] Examples of oligonucleotides include deoxyribonucleic acids (e.g., DNA, cDNA, etc.) or ribonucleic acids (e.g., RNA, miRNA, siRNA, piRNA, hnRNA, snRNA, sgRNA, esiRNA, shRNA, antisense oligonucleotides, lncRNA, etc.) polymers, or combinations of deoxyribonucleotides and ribonucleotides (e.g., DNA / RNA) polymers. The oligonucleotides of the present invention have a linear or circular structure and are in either single-stranded or double-stranded form. These terms should not be construed as limiting with respect to the length of the polymer and may include known analogs of natural nucleotides, as well as nucleotides chemically modified in the base, sugar, and / or phosphate moieties. Generally, analogs of a particular nucleotide have the same base pairing specificity. That is, an analog of A base pairs with T.

[0078] As used herein, the term "cellular exosomes" refers to a subset of extracellular vehicles (EVs) released by prokaryotes and eukaryotes. Exosomes typically exhibit a size range of about 40 - 160 nm (average about 100 nm) when of endosomal origin. Depending on the originating cell, exosomes may contain many components of the cell, including DNA, RNA, lipids, metabolites, as well as cytosolic proteins and cell surface proteins.

[0079] In one aspect, the pharmaceutical composition of the present invention further comprises one or more pharmaceutically acceptable carriers and / or diluents.

[0080] "Pharmaceutically acceptable carrier or diluent" generally means a carrier or diluent that is safe, non-toxic and useful for preparing a desired pharmaceutical composition, and includes carriers or diluents acceptable for human pharmaceutical use.

[0081] Such pharmaceutically acceptable carriers may be sterile liquids, such as water and oils, and the oils may include those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Aqueous physiological saline solutions and aqueous solutions of dextrose and glycerol can also be used as liquid carriers, especially for injection solutions.

[0082] The pharmaceutical composition may further contain one or more pharmaceutically acceptable salts, such as mineral acid salts, such as hydrochloride, hydrobromide, phosphate, sulfate, etc., and salts of organic acids, such as acetate, propionate, malonate, benzoate, etc. In addition, auxiliary substances, such as wetting agents or emulsifiers, pH buffering substances, gels or gelling substances, flavoring and odor-correcting agents, coloring agents, microspheres, polymers, suspending agents, etc. may also be present in the pharmaceutical composition. In addition, one or more other conventional pharmaceutical ingredients, such as preservatives, wetting agents, suspending agents, other surfactants, antioxidants, anti-caking agents, fillers, chelating agents, coating agents, chemical stabilizers, etc., may also be present, especially when the dosage form is in a reconstitutable form. Suitable exemplary ingredients include large crystalline cellulose, sodium carboxymethyl cellulose, polysorbate 80, phenylethyl alcohol, chlorobutanol, potassium sorbate, sorbic acid, sulfur dioxide, propyl gallate, parabens, ethyl vanillin, glycerol, phenol, parachlorophenol, gelatin, albumin, and combinations thereof. A thorough discussion of pharmaceutically acceptable excipients is available in REMINGTON’S PHARMACEUTICAL SCIENCES (Mack Pub. Co., New Jersey, 1991), which is incorporated herein by reference.

[0083] The pharmaceutical composition of the present invention can be administered orally, buccally, parenterally, nasally, topically or rectally. Preferably, the pharmaceutical composition is administered nasally or orally, more preferably by inhalation.

[0084] Also preferably, the pharmaceutical composition of the present invention is for use in the treatment of diseases. Any disease is contemplated in the present disclosure.

[0085] In one aspect, the pharmaceutical composition of the present invention is a vaccine. Vaccines typically contain attenuated organisms, inactivated organisms or killed organisms, or purified products (e.g., antigens) derived therefrom. Other examples of vaccines include nucleic acid vaccines containing antigens encoded by either DNA or RNA (e.g., mRNA). This nucleic acid vaccine is usually delivered using a viral vector (e.g., adenovirus) or a non-viral delivery system (e.g., electroporation or lipid nanoparticles).

[0086] The present disclosure provides, in some aspects, a method for the treatment and / or prevention of a disease, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) of the present invention or a pharmaceutical composition of the present invention.

[0087] As used herein, the term "therapeutically effective amount" means an amount of a compound of formula (I) that is high enough to significantly modify the symptoms and / or condition being treated in a favorable direction within the scope of sound medical judgment, but low enough to avoid serious side effects (with a reasonable risk / benefit ratio). The therapeutically effective amount of a compound of formula (I) is selected according to various factors including the type, species, age, weight, sex and medical condition of the patient, the severity of the condition being treated, the route of administration, and the renal and hepatic functions of the patient. A physician of ordinary skill in the art can readily determine and prescribe the effective amount of a compound of formula (I) necessary to prevent, counteract or halt the progression of the disease.

[0088] As used herein, the terms "treatment" or "treating" refer to any administration to a subject for the purpose of (i) inhibiting a disease, i.e., arresting the progression of clinical symptoms, and / or (ii) alleviating the disease, i.e., causing regression of clinical symptoms by any of the compositions, pharmaceutical compositions, therapeutic agents, compounds of formula (I), etc. of the present disclosure.

[0089] As used herein, the terms "prevention" or "preventing" refer to any administration to a subject for the purpose of preventing a disease of the present disclosure, i.e., preventing the onset of clinical symptoms of the disease, by any of the compositions, pharmaceutical compositions, therapeutic agents, compounds of formula (I), etc.

[0090] The present disclosure also provides a method for preparing a compound of formula (I). This method is described in the examples and · a step of sequentially and selectively functionalizing two primary alcohols of trehalose while maintaining other hydroxyl moieties unreacted, and · a step of allowing conjugation first to a fatty acid chain and then to a carboxylated side chain, wherein the carboxyl functional group is introduced in the form of a benzyl ester, which benzyl ester is removed in the last step of the procedure and includes.

[0091] The present disclosure further provides a method for preparing a polymer of a compound of formula (I), which method includes a step of self-condensing an activated trehalose derivative in the presence of a coupling agent. Examples of coupling agents are selected from the group including members of the carbodiimide family (e.g., DCC, EDCC, DIC, etc.), members of the benzotriazole family (e.g., TBTU, TATU, PyBOP, etc.), and members of the urea family (e.g., carbodiimidazole (CDI), etc.).

Examples

[0092] Example 1 Synthesis of succinyl-trehalose-fatty acids (Compounds 1 - 4) [Chemical formula]

[0093] Synthesis of trehalose-fatty acids: In a flame-dried round-bottom flask (20 mL) equipped with a magnetic stir bar, fatty acid (1.1 equiv) and TBTU (O-benzotriazol-1-yl-N,N,N’,N’-tetramethyluronium tetrafluoroborate) (1.1 equiv) were dissolved in anhydrous pyridine (6 mL). Then, trehalose (D-trehalose) (1 equiv) was poured into the reaction mixture, and stirring was continued at room temperature for 72 h under an argon atmosphere. Then, pyridine was removed under vacuum, and the resulting residue was purified using flash column chromatography with a solvent gradient of 5 - 20% methanol in EtOAc-DCM (1:1) to obtain the desired surfactant (yield 51%).

[0094] Synthesis of trehalose-fatty acids containing a succinyl functional group with a protecting group: In a flame-dried round-bottom flask equipped with a magnetic stir bar, monobenzyl succinate (1.1 equiv) and TBTU (1.1 equiv) were dissolved in anhydrous pyridine (2 mL / 0.1 g of trehalose derivative). Then, trehalose-fatty acid (1 equiv) was poured into the reaction mixture, and stirring was continued at room temperature (72 h) under an argon atmosphere. Then, pyridine was removed under vacuum, and the resulting residue was purified by flash column chromatography with a solvent gradient of 5 - 20% methanol in EtOAc-DCM (1:1) to obtain the desired surfactant (yield 30 - 40%).

[0095] Synthesis of succinyl-trehalose-fatty acids: Fatty acid - trehalose - succinate monobenzyl (1 equivalent) was dissolved in MeOH (5 mL) in a three - necked flask. After replacing the system atmosphere with argon, 10% Pd / C was added. Argon was removed from the system, and the reactant was flushed with hydrogen and returned (3 times). The reactant was vigorously stirred at room temperature for 2 - 3 hours (monitored by TLC). After completion of the reaction, the solution was filtered through a Celite pad, and the filter cake was washed with MeOH (10 mL). The filtrate was concentrated under reduced pressure to obtain the desired compound (95%).

[0096] Synthesis of trehalose polymers (Compound 5) [Chemical formula] Synthesis of trehalose containing a succinic acid functional group with a protecting group In a flame - dried round - bottom flask equipped with a magnetic stir bar, succinic acid monobenzyl ester (SAMBE) (1.1 equivalents) and 2 - (1H - benzotriazol - 1 - yl) - 1,1,3,3 - tetramethylaminium tetrafluoroborate (TBTU) (1.1 equivalents) described in Ballard, T.E.; Richards, J.J.; Wolfe, A.L.; Melander, C. Synthesis and Antibiofilm Activity of a Second - Generation Reverse - Amide Oroidin Library: A Structure - Activity Relationship Study. Chemistry - A European Journal 2008, 14, 10745 - 10761 were dissolved in anhydrous pyridine (2 mL / 0.1 g of trehalose). Then, trehalose (1 equivalent) was poured into the reaction mixture, and the mixture was stirred at room temperature for 42 hours under an argon atmosphere. Then, pyridine was removed under vacuum, and the resulting residue was purified by flash column chromatography using a solvent gradient of 9 - 20% methanol in EtOAc - DCM (1:1) to obtain the desired surfactant (yield 37%).

[0097] Synthesis of trehalose containing a succinic acid functional group Trehalose - succinic acid monobenzyl was dissolved in MeOH (1.5 mL / 0.1 g of trehalose derivative) in a three - necked flask. After replacing the system atmosphere with argon, 10% Pd / C (catalyst, about 10% molar) was added. Argon was removed from the system, and the reactant was flushed with hydrogen and returned (3 times). The reactant was vigorously stirred at room temperature for 2 - 3 hours (hr) (monitored by TLC). After completion of the reaction, the solution was filtered through a celite pad, and the filter cake was washed with MeOH (2× reaction volume). The filtrate was concentrated under reduced pressure to obtain the desired compound (83%).

[0098] Synthesis of protected trehalose polymers with n = 2 - 5 In a flame - dried round - bottom flask equipped with a magnetic stir bar, trehalose - succinic acid (1 equivalent) and TBTU (1 - 4 equivalents) were dissolved in anhydrous pyridine (2 mL / 0.1 g of trehalose derivative). This reaction mixture was stirred at room temperature for 42 hours under an argon atmosphere. Then, pyridine was removed under vacuum, and the resulting residue was purified by dialysis (100 - 500 MWCO).

[0099] Synthesis of succinyl - trehalose - unsaturated fatty acid

Chemical formula

[0100] Synthesis of succinyl - trehalose - unsaturated fatty acid: In a flame-dried round-bottom flask equipped with a magnetic stir bar, palmitoleic acid (1.2 equivalents) and TBTU (1.4 equivalents) were dissolved in anhydrous pyridine (3 mL / 0.1 g of palmitoleic acid). The reaction mixture was stirred at room temperature for 1 hour, after which TreSuc (1 equivalent) was added. Stirring was continued at room temperature for 96 hours under an argon atmosphere. Then, pyridine was removed under vacuum, and the resulting residue was purified by reverse-phase flash chromatography (H 2 O:MeCN 4:6→2:8). The fractions corresponding to the product were collected, concentrated in vacuo, and lyophilized to give the desired compound as a white solid (yield 15%).

[0101] Formulation Preparation Procedure The trehalose surfactant CnTreS solution was prepared by dissolving 5.6 mmol of the trehalose surfactant in 1 L of a commercially available 1X solution of PBS buffer. Then, a 6.3 mg / L antibody solution (provided in PBS (pH 6.5 - 6.7) containing 100 mM L-arginine) was diluted with the trehalose surfactant solution to a concentration of 1 mg / mL.

[0102] Spraying Procedure 500 μL to 8 mL of the antibody formulation was sprayed with a Briutcare mesh nebulizer (NEB-001). The flow rate was set to 0.4 mL / min, and the aerosol solution was collected in a 50 mL Falcon tube.

[0103] Lyophilization Procedure After placing 500 μL of the 1 mg / mL antibody formulation in a vial, it was rapidly frozen in liquid nitrogen. The frozen sample was placed directly into a lyophilizer. After 24 hours, the lyophilized sample was stored in a 4°C refrigerator for 7 days or 28 days. Then, the samples were reconstituted by the addition of MilliQ® water, and their stability was evaluated by DLS and cell-binding assays.

[0104] Stability Evaluation The stability of the antibody solution dissolved in the trehalose surfactant solution was tested over 10 days at 25°C. Antibody aggregation and conformation were measured by dynamic light scattering DLS using a nano-ZS (Malvern Panalytical) and a nano-DSF instrument (NanoTemper Technology), respectively, and the Fab ability to interact with the antigen functional group was evaluated by an ELISA assay developed for this study.

[0105] Cell binding assay Functional stability was evaluated by studying its ability to bind to antigens on the cell membrane. Wild-type HEK cells and HEK cells expressing SARS-CoV-2 spike (D614) were cultured in RPMI medium containing 10% fetal bovine serum, 100 U / ml penicillin, and 100 μg / ml streptomycin. Both cell lines were incubated with 50 μL of a 0.5 μg / mL antibody formulation at 4°C for 30 minutes. The cells were washed in PBS and incubated with alexa Fluor 488 conjugated mouse anti-human Fc IgG diluted 1 / 500 in PBS for 15 minutes at 4°C in the dark. The cells were washed and the fluorescence-activated cells were analyzed by FACS using a CANTO II (BD Bioscience, USA).

[0106] Colloidal stability evaluation Antibody aggregation was evaluated by dynamic light scattering (DLS) operated at 25°C. Measurements were taken at different time points during storage at 25°C and immediately after spraying.

[0107] The average hydrodynamic diameter by monomer intensity was distributed at d = 10 nm, and the aggregates were distributed at d > 10 nm.

[0108] Cytotoxicity assay The cytotoxicity of the trehalose surfactant was evaluated using the NIH-3T3 cell line. Trehalose surfactant solutions in the range of 0.0112 to 11.2 mM were prepared by diluting a stock solution in cell culture medium (DMEM supplemented with 10% fetal bovine serum, 2 mM glutamine, and 1% penicillin / streptomycin). The cells were incubated with the diluted surfactant at 37 °C for 48 hours, and a solution of MTS / PMS (MTS CellTiter96 AQueous assay) was added according to the supplier's instructions. The cells were then incubated at 37 °C for 3 hours, and the absorbance was measured at 490 nm using a MultiSkan microplate reader (Thermo Fisher Scientific, USA). The cell viability was plotted against the surfactant concentration, and the cytotoxic dose median (CD50) was calculated.

[0109] Example 2 Synthesis The trehalose surfactant is produced from biocompatible, non-toxic, and inexpensive raw materials (saturated or unsaturated fatty acids, trehalose, and succinic acid) available from earth-friendly sources. These can be synthesized from unprotected trehalose by an asymmetric strategy. These can be easily degraded and metabolized by the human body through enzymatic and hydrolytic cleavage of the ester bond. In one embodiment, the excipient contains a succinic acid functional group to enhance the interaction between the biomolecule and the excipient via electrostatic interaction. In another aspect, the excipient contains two units of trehalose to increase the hydrophilic portion of the surfactant, improve the solubility of the biomolecule / surfactant complex in aqueous solution, and enhance the stability of the biomolecule through the formation of hydrogen bonds.

[0110] Storage stability The inventors have shown that trehalose surfactants enhance biomolecule solubility through the formation of drug-surfactant colloidal complexes while improving their storage-life stability. Storage-life stability was monitored over 4 days by evaluation of aggregate formation by DLS (Figure 1). IgG at 1 mg / mL in PBS showed aggregation after 4 days, with or without excipients.

[0111] Stability during spraying The use of trehalose surfactants in antibody formulations reduces aggregation and conformational changes during spraying. The stability of the antibody after spraying was evaluated by DLS and nano-DSF to assess aggregate formation and conformational changes after spraying in freshly prepared formulations, respectively. As shown in Figure 2, spraying in PBS showed only 9.7% aggregates of the total AUC. Spraying appears to dramatically accelerate aggregation compared to the rate of aggregation during storage (only 3.5% in PBS after 10 days) observed during storage at 25°C.

[0112] Aggregation was observed with 1 mM C8Tre1 or C16Tre1S (about 5% of the total volume-weighted AUC). Aggregates were avoided when the formulation contained C8Tre1 or C16Tre1S at a minimum concentration of 5.6 mM. This data is in good agreement with the storage-life stability tests.

[0113] The colloidal stability of the antibody after spraying was also evaluated by Nano-DSF. The difference between the scattering values at 70°C (the highest level of aggregation) and 20°C (the lowest level of aggregation) was calculated for each sample before and after spraying (Figure 3).

[0114] The results showed that for most samples, similar results were obtained before and after spraying. However, the 1 mM trehalose surfactant showed aggregates that were probably generated by spraying. This data is in good agreement with the DLS experiment showing aggregation at the same concentration. C8Tre1 and C16Tre1S prevented antibody aggregation when used at a minimum concentration of 5.6 mM. The lack of change before and after spraying suggested that the same proportion of aggregates was present before and after heating in the nano-DESF device.

[0115] Stability during lyophilization: The addition of 5.6 mM C16Tre1S and C8Tre1S trehalose surfactants as cryoprotectants was evaluated in 1 mg / mL antibody formulations. Samples were first rapidly frozen in liquid nitrogen and then dried for 24 hours. The lyophilized samples were stored in a refrigerator for 0, 7, or 28 days and then reconstituted by adding MilliQ® water. The antibody's conformation and functionality after this process were evaluated by DLS and cell binding assays, respectively. The results were compared with antibodies formulated in PBS only, standard cryoprotectants such as trehalose (5 wt% and 5.6 mM), and antibodies formulated with Tween80 (5.6 mM).

[0116] The DLS results (Figure 4) showed that no aggregation was observed for the formulation containing C16Tre1S, and this lyophilized antibody remained stable when stored at 4 °C for 28 days. Equivalent results were obtained with two standard cryoprotectants (5.6 mM Tween80 and 5 wt% trehalose). However, when a small amount of trehalose was used (5.6 mM), aggregation was observed as in the case of the sample containing PBS only. For the sample containing C8Tre1S, aggregation was observed after lyophilization, and the amount of aggregates increased with storage time, suggesting that C8Tre1S is cryoprotective but does not stabilize the antibody during storage.

[0117] Next, the ability of the antibody to bind to its antigen after lyophilization was evaluated in vitro using SARS-CoV-2 spike-expressing HEK293 cells (Figure 6). After antibody treatment, the cells were labeled with Alexafluor488. When the antibody was formulated with C16TreS, a high fluorescence signal was observed for all HEK293 cells expressing the spike protein. Equivalent results were obtained using the same concentration of the standard cryoprotectant Tween80. However, significant decreases in antibody binding were observed with formulations using trehalose (5 wt% and 5.6 mM), C8TreS (5.6 mM), and surfactant-free formulations. When the reconstituted antibody formulation containing Tween80 and C16TreS was subjected to a spray challenge, the antibody was able to bind to the spike protein at the same level as before spraying. These results suggest that the C16Tre1S trehalose surfactant stabilizes the antibody to an extent equivalent to the standard cryopreservative Tween80 during lyophilization, during 28 days of storage when lyophilized, and during the spraying process.

[0118] Cytotoxicity The cytotoxicity of all surfactants used in this study was evaluated for NIH-3T3 cells 48 hours later in an MTS cell viability assay. C8Tre1 showed a CD50 of 5.4 mM, and for C14Tre1, C14Tre1S, or C16Tre1S, they were 0.30 mM, 0.84 mM, and 0.24 mM, respectively. It seems that the longer the alkyl chain, the higher the cytotoxicity.

[0119] Since the CD50 was 5.4 mM for C8Tre1S, but greater than 11.2 mM for C8Tre1, succinylation of trehalose also seems to increase toxicity. Anionic surfactants are generally considered to be more toxic than nonionic surfactants.

Claims

1. Compound of formula I 【Chemistry 1】 or its salt, stereoisomer, polymorph, or mixture of stereoisomers, During the ceremony, R １ is selected from alkyl groups which may be substituted with -OH,OH, or R via oxygen ３ It is connected, R ２ is selected from alkyl groups which may be substituted with -OH,OH, or R via oxygen ３ It is connected, R ３ is selected from alkyl groups which may be substituted with -OH,OH, or R via oxygen １ Or R ２ It is connected, R ４ is selected from alkyl groups which may be substituted with -OH or -OH groups. X １ is, independently, selected from -(C=O) and -(C=O)-A-(C=O), where A is alkyl, alkenyl, alkynyl or PEG, X ２ These are independently selected from H or -(C=O), Z １ is -OH, X ２ If C = O, then Z ２ These are independently selected from alkyl, alkenyl, alkynyl, PEG, succinyl, -NH alkyl, -NH alkenyl, -NH alkynyl, NH-PEG, -S alkyl, -S alkynyl, -O alkyl, -O alkenyl, and -O alkynyl. X ２ If H, then Z ２ It does not exist. n is an integer between 1 and 20. A compound of formula I, or its salt, stereoisomer, polymorph, or mixture of stereoisomers.

2. R １ is -OH, R ２ R is transmitted via oxygen. ３ It is connected, R ４ OH is, X １ is -(C=O)-CH2-CH2-(C=O), Z １ is -OH, X ２ is -(C=O), Z ２ is alkyl, alkenyl, or absent. The compound according to claim 1.

3. The aforementioned compound, Table 1(1) 【Table 1(2)】 A compound according to claim 1, selected from the group consisting of the following.

4. A composition comprising a compound of formula I as described in any one of claims 1 to 3.

5. The composition according to claim 4, wherein the compound of formula I is a surfactant or an excipient.

6. A pharmaceutical composition comprising the compound of formula I as described in claim 1.

7. A pharmaceutical composition comprising the compound of formula I described in claim 2.

8. A pharmaceutical composition comprising the compound of formula I described in claim 3.

9. The pharmaceutical composition according to claim 6, further comprising a therapeutically effective amount of pharmaceutical agent.

10. The pharmaceutical composition according to claim 7, further comprising a therapeutically effective amount of pharmaceutical agent.

11. The pharmaceutical composition according to claim 8, further comprising a therapeutically effective amount of pharmaceutical agent.

12. The pharmaceutical composition according to claim 9, wherein the pharmaceutical agent is selected from the group comprising chemical compounds, peptides, lipids, oligonucleotides, cell exosomes, and one or more combinations thereof.

13. The pharmaceutical composition according to claim 10, wherein the pharmaceutical agent is selected from the group comprising chemical compounds, peptides, lipids, oligonucleotides, cell exosomes, and one or more combinations thereof.

14. The pharmaceutical composition according to claim 11, wherein the pharmaceutical agent is selected from the group comprising chemical compounds, peptides, lipids, oligonucleotides, cell exosomes, and one or more combinations thereof.

15. The pharmaceutical composition according to claim 12, wherein the peptide is selected from the group comprising antibodies, antigen-binding proteins, and combinations thereof.

16. The pharmaceutical composition according to claim 13, wherein the peptide is selected from the group comprising antibodies, antigen-binding proteins, and combinations thereof.

17. The pharmaceutical composition according to claim 14, wherein the peptide is selected from the group comprising antibodies, antigen-binding proteins, and combinations thereof.

18. The pharmaceutical composition according to claim 12, wherein the oligonucleotide is selected from the group comprising deoxyribonucleic acid, ribonucleic acid, and combinations thereof.

19. The pharmaceutical composition according to claim 13, wherein the oligonucleotide is selected from the group comprising deoxyribonucleic acid, ribonucleic acid, and combinations thereof.

20. The pharmaceutical composition according to claim 14, wherein the oligonucleotide is selected from the group comprising deoxyribonucleic acid, ribonucleic acid, and combinations thereof.

21. A pharmaceutical composition according to any one of claims 6 to 20 for use in the treatment of a disease.

22. The pharmaceutical composition according to any one of claims 6 to 20, which is administered orally, intraoral, parenterally, nasally, topically, or rectally.

23. The pharmaceutical composition according to claim 22, wherein the pharmaceutical composition is administered by inhalation, either nasally or orally.

24. A pharmaceutical composition according to any one of claims 6 to 20, which is a vaccine.

25. A method for preparing a compound of formula I according to any one of claims 1 to 3, A process in which the two primary alcohols of trehalose are functionalized sequentially and selectively while the other hydroxyl moieties remain unreacted, A step in which conjugation is first carried out to a fatty acid chain and then to a carboxylated side chain, wherein the carboxyl functional group is introduced in the form of a benzyl ester, and the benzyl ester is removed in the final step of the procedure. Methods that include...

26. A method for preparing a polymer of a compound of formula I according to any one of claims 1 to 3, comprising the step of self-condensing an activated trehalose derivative in the presence of a coupling agent.

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