Saponin-based immunostimulants, pharmaceutical compositions containing the immunostimulants, and their therapeutic uses

Abstract

The present disclosure provides compounds containing modified saponin compounds, pharmaceutical compositions containing modified saponin compounds, methods of using modified saponin compounds and pharmaceutical compositions, and methods of manufacturing modified saponin compounds. The compounds and pharmaceutical compositions of the present disclosure can be used in combination with one or more other therapeutic agents for treating viral infections and other diseases.

Description

Technical Field

[0001] Claim of Priority of Related Applications This application claims priority to U.S. Provisional Application No. 63 / 364,897, filed May 18, 2022, entitled "SAPONIN-BASED IMMUNOSTIMULANTS, PHARMACEUTICAL COMPOSITION COMPRISING SAID IMMUNOSTIMULANTS, THERAPEUTIC USE THEREOF", the entire disclosure of which is hereby incorporated by reference in its entirety.

[0002] Statement Regarding Funding Provided by the U.S. Government This invention was made with government support under Contract No. R01 GM120159 awarded by the National Institutes of Health. The government has certain rights in this invention.

Background Art

[0003] A vaccine adjuvant is a substance used together with a vaccine to enhance the host immune response against specific antigen(s) introduced by the vaccine (Brunner et al., (2010) Immunol. Lett. 128:29-35, Kensil et al., (2004) Frontiers Biosci. 9:2972-2988, Leroux-Roels G. (2010) Vaccine 28(Suppl 3):C25-36, Sharp & Lavelle (2012) Development Therapeutic Agents Handbook John Wiley & Sons, Inc; pp.533-546, Wang W. (2011) World J. Vaccines 1:33-78, Weeratna & McCluskie (2011) Recent Advan. Vaccine Adjuvants.pp.303-322, Cox & Coulter (1997) Vaccine 15:248-256, Klebanoff et al., (2010) Immunol. Rev. 239:27-44, Plotkin SA. (2005) Nat. Med. 11:S5-S11, Rappuoli & Aderem (2011) Nature 473:463-469, Kensil et al., (2005) Vaccine Adjuvants: Immunological and Clinical Principles Humana Press Inc.pp.221-234).

[0004] Vaccine adjuvants also modulate the immune system to a desirable response against a particular pathogen. For example, QS-21, a mixture of two isomers, is an FDA-approved adjuvant known for its ability to enhance a balanced Th1 / Th2 response in antigen-specific CTL production and is beneficial for vaccines against intracellular pathogens and cancer (Ragupathi et al., (2011) Expert Rev. Vaccines 10:463-470, Deng et al., (2008) Angew Chem. Int. 47:6395-6398, Kensil CR. (1996) Critical Revs. Therap. Drug Carrier Systs. 13:1-55, Kensil et al., (1991) J. Immun. 146:431-437). It has potential for a wide range of clinical applications and thus there is high demand (Kensil et al., (1991) J. Immun. 146:431-437). The supply of QS-21 is very limited. The natural product is isolated from the bark of Quillaja saponaria Molina (QS), an evergreen tree native to the warm central region of Chile. However, even under current demand, overcollection from natural sources has ecological and economic consequences. Furthermore, the abundance of QS-21 in QS bark extracts is low and its isolation is laborious (Kensil et al., (1991) J. Immun. 146:431-437, Ragupathi et al, (2010) Vaccine 28:4260-4267; Wang et al., (2005) J. Am. Chem. Soc. 127:3256-3257). QS-21 also has problems of chemical instability due to two hydrolytically labile ester moieties that complicate its formulation, and its toxicity that limits its dosage also prevents reaching full efficacy.

[0005] Derivatization of Momordica saponins (MS) I and II has been shown to be a potentially viable approach to achieve a practical alternative to QS-21 (Wang et al., (2019) J. Med. Chem. 62:9976-9982). MS I and II are isolated from the seeds of the perennial Momordica cochinchinensis Spreng (MC) that grows in China and Southeast Asia. Derivatization of MS I / II with C3 glucuronic acid may result in an immunostimulant having an adjuvant effect similar to QS-21.

Summary of the Invention

[0006] The present disclosure provides compounds containing modified saponin compounds, pharmaceutical compositions containing modified saponin compounds, methods of using modified saponin compounds and pharmaceutical compositions, and methods of making modified saponin compounds, etc.

Means for Solving the Problems

[0007] The present disclosure provides a modified saponin having the following formula:

[0008]

Chemical Formula

[0009] (In the formula, q1 can be H or OH, q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a constituent of an acetal group, or an imine group, f3 and f4 are each independently OH, or acetyl, or C3 and C4 of a fuocsyl unit, and f3 and f4 can form a cyclic ketal ring or a cyclic carbonate ester, f5 is selected from H, a methyl group, -CH2OH group, R4-NR5-C(O)-, R4-O-C(O)-, or R4-O-CH2-, R4 and R5 are each independently a structure R6[(CX2) 0-20 O 0-1(CY2) 0-20 0-20 is a straight chain having X and Y are each independently H or a halogen atom, R6 is COOR7, C(O)NR7R8, NR7R8, or OR7, R7 is Ar[(CZ2) 0-20 O 0-1 (CL2) 0-20 0-20 and R8 is H or alkyl, Z and L are each independently H or a halogen atom, Ar is a substituted or unsubstituted aromatic system, ga5 is R 14 -NR 15 -C(O)-, R 14 -O-C(O)-, or R 14 -O-CH2- selected from, R 14 and R 15 are each independently a straight chain having the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 0-20 and X1 and Y1 are each independently H or a halogen atom, R 16 is COOR 17 , C(O)NR 17 R 18 , NR 17 R 18 , or OR 17 , R 17 is Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 0-20 and R 18 is H or alkyl, Z1 and L1 are each independently H or a halogen atom, Ar2 is a substituted or unsubstituted aromatic group, r3 is H, monosaccharide, disaccharide, or trisaccharide, ​​​​x3 is H, a monosaccharide (excluding xylose), or a disaccharide, and ga3 is H, a monosaccharide, or a disaccharide).

[0010] The present disclosure provides a modified saponin that may have the following formula I:

[0011]

Chemical formula

[0012] The present disclosure provides a modified saponin that can be selected from the group consisting of the following formulas 2D - 2H.

[0013]

Chemical formula

[0014] The present disclosure provides a pharmaceutical composition for treating a condition, comprising a therapeutically effective amount of a modified saponin having the following formula:

[0015]

Chemical formula

[0016] (wherein, q1 is H or OH, q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a component of an acetal group, or an imine group, f3 and f4 are each independently OH, or acetyl, or C3 and C4 of a fuocsyl unit, and f3 and f4 can form a cyclic ketal ring or a cyclic carbonate ester, f5 is selected from H, a methyl group, -CH2OH group, R4 - NR5 - C(O)-, R4 - O - C(O)-, or R4 - O - CH2-, R4 and R5 are each independently a straight chain having the structure R6[(CX2) 0-20 O 0-1 (CY2) 0-20 0-20 and X and Y are each independently H or a halogen atom, R6 is COOR7, C(O)NR7R8, NR7R8, or OR7, R7 is Ar[(CZ2) 0-20 O 0-1 (CL2) 0-20 0-20 and R8 is H or alkyl, Z and L are each independently H or a halogen atom,​​ Ar is a substituted or unsubstituted aromatic system, ga5 is R 14 -NR 15 -C(O)-, R 14 -O-C(O)-, or R 14 -O-CH2-, and is selected from R 14 and R 15 each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 0-20 and is a straight chain, X1 and Y1 each independently are H or a halogen atom, R 16 is COOR 17 , C(O)NR 17 R 18 , NR 17 R 18 , or OR 17 and R 17 is Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 0-20 and R 18 is H or alkyl, Z1 and L1 each independently are H or a halogen atom, Ar2 is a substituted or unsubstituted aromatic group, r3 is H, a monosaccharide, a disaccharide, or a trisaccharide, x3 is H, a monosaccharide (excluding xylose), or a disaccharide, and ga3 is H, a monosaccharide, or a disaccharide).

[0017] The present disclosure provides a pharmaceutical composition comprising a therapeutically effective amount of a modified saponin having the following formula I for treating a condition:

[0018]

Chemical formula

[0019] (wherein, q1 is H or OH, q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a constituent of an acetal group, or an imine group, ga5 is R 14 -NR 15 -C(O)-, R 14 -O-C(O)-, or R 14 -O-CH2- is selected from, R 14 and R 15 are each independently a straight chain having the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 0-20 where X1 and Y1 are each independently H or a halogen atom, X1 and Y1 are each independently H or a halogen atom, R 16 is COOR 17 , C(O)NR 17 R 18 , NR 17 R 18 , or OR 17 and R 17 is Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 0-20 where R 18 is H or alkyl, Z1 and L1 are each independently H or a halogen atom, and Ar2 is a substituted or unsubstituted aromatic group).

[0020] The present disclosure provides a therapeutically effective amount of a modified saponin having one of the following formulas 2D-H for treating a condition.

[0021]

Chemical formula

[0022] ​​The present disclosure provides a pharmaceutical composition that may also include at least one immunogen, a pharmaceutically acceptable carrier, and at least one therapeutic agent (e.g., at least one cancer therapeutic agent), among other things as described above. The pharmaceutical composition may be formulated for administration to an animal or human subject.

[0023] Further aspects of the present disclosure will be more readily understood upon consideration of the detailed description of its various embodiments, which is set forth below when taken in conjunction with the accompanying drawings.

Brief Description of the Drawings

[0024]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5A

Figure 5B

Figure 5C

Figure 6A

Figure 6B

Figure 6C

Mode for Carrying Out the Invention

[0025] The present disclosure is not limited to the specific embodiments described and can, of course, be different. The technical terms used herein are not intended to limit the scope of the present disclosure, which is only limited by the appended claims, and are not intended to be limiting as they are only for the purpose of describing specific embodiments.

[0026] When ranges of values are provided, each intervening value, to one tenth of the unit of the lower limit, between the upper and lower limits of the range, and any other stated value or intervening value within the stated range, are included within the disclosure, unless the context clearly dictates otherwise. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are included within the disclosure, subject to any expressly excluded limitations within the stated range. When the stated range includes one or both of the limits, ranges excluding one or both of those included limits are also included in the disclosure.

[0027] Embodiments of the disclosure will employ techniques within the scope of the art, such as medicine, organic chemistry, biochemistry, molecular biology, pharmacology, etc., unless otherwise indicated. Such techniques are well described in the literature.

[0028] The following examples are set forth to provide those skilled in the art with a complete disclosure and description of how to perform the methods disclosed and claimed herein, as well as how to use the compositions and compounds. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. Unless otherwise indicated, parts are parts by weight, temperature is in °C, and pressure is at or near atmospheric pressure. Standard temperature and pressure are defined as 20 °C and 1 atmosphere.

[0029] Before the embodiments of the present disclosure are described in detail, it will be understood that the present disclosure is not limited to specific materials, reagents, reactants, manufacturing processes, dimensions, frequency ranges, or applications, etc., and can vary, unless otherwise indicated. It should also be understood that the terminology used herein is for the purpose of describing only specific embodiments and is not intended to be limiting. In the present disclosure, steps can also be performed in different orders if this is logically possible. Embodiments of the present disclosure can also be applied to additional embodiments that include measurements beyond those described in the examples herein, which are not intended to be limiting. Embodiments of the present disclosure can further be combined with or integrated with other measurement techniques beyond those described in the examples herein, which are not intended to be limiting.

[0030] It should be noted that when used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a support" includes a plurality of supports. In this specification and the following claims, several terms are referred to and are defined to have the following meanings unless the contrary intention is apparent.

[0031] Each application and patent cited in the text, and each document or reference cited in each application and patent (including during the prosecution of each issued patent; "application cited documents"), and each PCT and foreign application or patent corresponding to and / or claiming priority from any of these applications and patents, and each document cited or referenced in each application cited document are hereby expressly incorporated by reference into this specification. Further, each document or reference cited in the text, the list of reference documents before the claims, or the text itself, and each of these documents or references ("reference documents cited in this specification"), and each document or reference cited in each reference document cited in this specification (including any manufacturer's specifications, instructions, etc.) are hereby expressly incorporated by reference into this specification.

[0032] Before describing various embodiments, the following definitions are provided and should be used unless otherwise indicated.

[0033] Definitions "Substituted" or "substitution" includes the implicit condition that such substitution follows the valences of the substituted atoms and substituents, and it is understood that the substitution results in a stable compound, i.e., a compound that does not undergo spontaneous transformation such as rearrangement, cyclization, elimination, etc.

[0034] It will be understood by those skilled in the art that, where appropriate, the substituted moiety itself may be further substituted. For example, substituents on a substituted alkyl can include halogen, hydroxy, nitro, thiol, amino, azido, imino, amide, phosphoryl (including phosphonates and phosphinates), sulfonyl (including sulfates, sulfonamides, sulfamoyl and sulfonates), and silyl groups, as well as ether, alkylthio, carbonyl (including ketones, aldehydes, carboxylates, and esters), -CF3, -CN, etc., in both substituted and unsubstituted forms. Cycloalkyl can be substituted in a similar manner.

[0035] As used herein, the term "acyl" alone or in combination means a carbonyl or thiocarbonyl group bonded to a radical selected from, for example, optionally substituted hydride, alkyl (e.g., haloalkyl), alkenyl, alkynyl, alkoxy (including "acyloxy" such as acetyloxy, butyryloxy, iso-butyryloxy, phenylacetyloxy, berizoyloxy, p-methoxybenzoyloxy, and alkoxyalkyl and haloalkoxy), aryl, halo, heterocyclyl, heteroaryl, sulfonyl (e.g., allylsulfinylalkyl), sulfonyl (e.g., alkylsulfonylalkyl), cycloalkyl, cycloalkenyl, thioalkyl, thioaryl, amino (e.g., alkylamino or dialkylamino), and aralkoxy. Exemplary examples of "acyl" radicals are formyl, acetyl, 2-chloroacetyl, 2-bromacetyl, benzoyl, trifluoroacetyl, phthaloyl, malonyl, and nicotinyl. As used herein, the term "acyl" refers to the group -C(O)R 26 wherein R 26 is hydrogen, alkyl, cycloalkyl, cycloheteroalkyl, aryl, arylalkyl, heteroalkyl, heteroaryl, and heteroarylalkyl. Examples include, but are not limited to, formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl, and beozylcarbonyl.

[0036] As used herein, the term "adjuvant molecule" refers to a surface protein capable of inducing an immune response in a host. In certain embodiments, the adjuvant molecule is an adjuvant molecule in "membrane-anchored form", which indicates that the adjuvant molecule has been engineered to contain a signal peptide (SP) and a membrane-anchoring sequence to direct protein transport and membrane orientation. Thus, in embodiments, the adjuvant molecule in membrane-anchored form is a recombinant protein comprising a portion of the protein fused to the SP and the membrane-anchoring sequence.

[0037] As used herein, the terms "administering" and "administration" refer to introducing a composition of the present disclosure (e.g., a vaccine, an adjuvant, or an immunogenic composition) to a subject. As used herein, "administering" can refer to administration by oral, topical, intravenous, subcutaneous, transdermal, transepidermal, intramuscular, intra-articular, parenteral, intra-arterial, intradermal, intraventricular, intraosseous, intraocular, intracranial, intraperitoneal, intralesional, intranasal, intracardiac, intra-articular, intracorporeal, intrathecal, intravitreal, intracerebral, and intraventricular, intratympanic, intracochlear, rectal, intravaginal, by inhalation, by catheter, by stent, or via other devices for administering the composition either actively or passively (e.g., by diffusion) into the reservoir or perivascular space and adventitia. The preferred route of administration of the vaccine composition is intravenous.

[0038] As used herein, the term "alkoxyl" or "alkoxyalkyl" refers to an alkyl-O-group where the alkyl is as described above. As used herein, the term "alkoxyl" refers to, for example, linear, branched or cyclic, saturated or unsaturated oxo-hydrocarbon chains including methoxyl, ethoxyl, propoxyl, isopropoxyl, butoxyl, t-butoxyl, and pentoxyl, containing C 1-20 can refer to.

[0039] As used herein, the term "alkyl," alone or in combination with any other terms such as "thioalkyl" and "arylalkyl," means a monovalent saturated hydrocarbon radical that can be linear (i.e., straight-chain) or branched-chain. Alkyl radicals for use in the present disclosure generally contain from about 1 to 20 carbon atoms, particularly about 1 to 10, 1 to 8, or 1 to 7 carbon atoms, more specifically about 1 to 6, or 3 to 6 carbon atoms. Exemplary alkyl radicals include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, isopentyl, amyl, sec-butyl, tert-butyl, tert-pentyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, n-dodecyl, n-tetradecyl, pentadecyl, n-hexadecyl, heptadecyl, n-octadecyl, nonadecyl, eicosyl, docosyl, and n-tetracosyl, among other branched variations. In certain embodiments of the present disclosure, the alkyl radical is a C1-C6 lower alkyl selected from the group consisting of or comprising methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, isopentyl, amyl, tributyl, sec-butyl, tert-butyl, tert-pentyl, and n-hexyl. The alkyl radical may optionally be substituted with substituents as defined herein at a position that does not significantly interfere with the preparation of the compounds of the present disclosure and does not significantly reduce the effectiveness of the compounds. In certain embodiments of the present disclosure, the alkyl radical is substituted with 1 to 5 substituents including halo, lower alkoxy, lower aliphatic, substituted lower aliphatic, hydroxy, cyano, nitro, thio, amino, keto, aldehyde, ester, amide, substituted amino, carboxyl, sulfonyl, sulfuryl, sulfenyl, sulfate, sulfoxide, substituted carboxyl, halogenated lower alkyl (e.g., CF3), halogenated lower alkoxy, hydroxycarbonyl, lower alkoxycarbonyl, lower alkylcarbonyloxy, lower alkylcarbonylamino, alicyclic-substituted cycloaliphatic, or aryl (e.g., phenylmethylbenzyl)), heteroaryl (e.g., pyridyl), and heterocyclic (e.g., piperidinyl, morpholinyl).The substituents on the alkyl group can themselves be substituted.

[0040] The Ar (e.g., Ar1, Ar2, etc.) group is a group such as an aromatic or aryl group. As used herein, "aryl" refers to a C5-C 20 member aromatic ring system, heterocyclic ring system, fused aromatic ring system, fused heterocyclic ring system, biaromatic ring system, or biheterocyclic ring system. In one aspect, "aryl" can be a 5-, 6-, 7-, 8-, 9-, and 10-membered monocyclic aromatic group that can contain 0 to 4 heteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, and pyridazine and pyrimidine and other corresponding functional groups. These aryl groups having heteroatoms in the ring structure can also be referred to as "aryl heterocycles" or "heteroaromatic compounds". The aromatic ring can be substituted at one or more ring positions with, but not limited to, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino (or quaternized amino), nitro, sulfhydryl, imino, amide, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, -CF3, -CN; and one or more substituents including combinations thereof.

[0041] Also, the term "aryl" refers to a polycyclic ring system (C5-C 30) and at least one of the rings is aromatic. For example, the other ring(s) may be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, and / or heterocyclyl.Examples of heterocyclic rings include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thieno[2,3-b]thiazolyl, thieno[2,3-b]oxazolyl, thieno[2,3-b]imidazolyl, thiophenyl, and xanthenyl.One or more rings may be substituted as defined above for "aryl".

[0042] As used herein, the term "antibody" refers to polyclonal and monoclonal antibody preparations, as well as hybrid antibodies, modified antibodies, F(ab’)2 fragments, F(ab) fragments, Fv fragments, single domain antibodies, chimeric antibodies, humanized antibodies, and functional fragments thereof that exhibit the immunological binding properties of the parent antibody molecule.

[0043] As used herein, the term "antibody" further refers to immunoglobulins that specifically bind to the specific spatial and polar organization of another molecule and are thereby defined as complementary. Antibodies can be monoclonal antibodies, polyclonal antibodies, or recombinant antibodies and can be prepared by techniques well known in the art such as immunization of a host and collection of serum (polyclonal), or by preparing continuous hybrid cell lines and collecting the secreted protein (monoclonal), or by cloning and expressing a nucleotide sequence or a mutated version thereof that encodes at least the amino acid sequence necessary for the specific binding of a natural antibody. Antibodies can include intact immunoglobulins or fragments thereof, and such immunoglobulins include various classes and isotypes such as IgA, IgD, IgE, IgG1, IgG2a, IgG2b, and IgG3, IgM, IgY. Examples of such fragments include Fab, Fv, and F(ab’)2, Fab’, and scFv. In addition, aggregates, polymers, and conjugates of immunoglobulins or fragments thereof can be used where appropriate as long as the binding affinity for a particular molecule is maintained.

[0044] As used herein, the term "antigen" refers to a molecule having one or more epitopes that stimulate the host immune system to generate a secretory, humoral, and / or cellular antigen-specific response, or a DNA molecule capable of producing such an antigen in a vertebrate. The term is also used interchangeably with "immunogen". For example, a particular antigen can be a complete protein, a portion of a protein, a peptide, a fusion protein, a glycosylated protein, and combinations thereof. For use with the compositions of the present disclosure, one or more PvDBPII antigens (native proteins or protein fragments) may be provided directly or as part of a recombinant nucleic acid expression system to provide an antigenic PvDBPII product that elicits a host immune response.

[0045] As used herein, the term "antigenic component" refers to a component derived from an organism that is capable of stimulating an immune response in animals, preferably mammals including mice and humans. An antigenic component may be an immunogenic substance. Antigenic components can include intracellular components, including organelles, membranes, proteins, lipids, glycoproteins, and other components derived from an organism. Antigenic components can be derived from an entire organism, such as an entire parasite, or a part of an organism, such as a cell or tissue of an organism. Also, subsets of proteins can be purified and recombined, for example, by size fractionation or affinity purification.

[0046] As used herein, the terms “sugar” and “saccharide” refer to polyhydroxyaldehydes, polyhydroxyketones, and their derivatives. Examples of this term include monosaccharides such as erythrose, arabinose, allose, altrose, glucose, mannose, threose, xylose, gulose, idose, galactose, talose, aldhexose, fructose, ketohexose, ribose, and aldopentose. This term also includes carbohydrates composed of monosaccharide units, including disaccharides, oligosaccharides, or polysaccharides. Examples of disaccharides are sucrose, lactose, and maltose. Oligosaccharides generally contain 3 to 9 of those monosaccharides, and polysaccharides contain more than 10 monosaccharide units. Sugars can be members of the D or L series and can include amino sugars, deoxy sugars, and their uronic acid derivatives. In embodiments of the present disclosure where the carbohydrate is a hexose, the hexose is glucose, galactose, or mannose, or a substituted hexose sugar residue such as an amino sugar residue such as hexosamine, galactosamine, glucosamine, particularly D-glucosamine (2-amino-2-deoxy-D-glucose) or D-galactosamine (2-amino-2-deoxy-D-galactose). Exemplary pentose sugars include arabinose, fucose, and ribose. Sugar residues from 1,1-linkage, 1,2-linkage, 1,3-linkage, 1,4-linkage, 1,5-linkage, or 1,6-linkage can be linked to the compounds of the present disclosure. The linkage can be through an oxygen atom of the disclosed compound. The oxygen atom can be replaced one or more times by a --CH2-- or --S-- group.

[0047] As used herein, the term “carboxyl” alone or in combination refers to -C(O)OR 25 - or -C(-O)OR 25 wherein R 25is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, amino, thiol, aryl, heteroaryl, thioalkyl, thioaryl, thioalkoxy, heteroaryl, or heterocyclic, and these may be optionally substituted. In aspects of the present disclosure, the carboxyl group is in an esterified form and may contain a lower alkyl group as the esterifying group. In certain aspects of the present disclosure, -C(O)OR 25provides an ester or amino acid derivative. The esterified form is also sometimes referred to herein specifically as a "carboxylic acid ester". In aspects of the present disclosure, "carboxyl" may be substituted, and in particular may be substituted with allyl optionally substituted with one or more of amino, amine, halo, alkylamino, aryl, carboxyl, or heterocyclic. Examples of carboxyl groups include tert. alkoxycarbonyl such as methoxycarbonyl, butoxycarbonyl, tert-butoxycarbonyl, and benzyloxycarbonyl, methoxybenzyloxycarbonyl, diphenylmethoxycarbonyl, 2-bromoethoxycarbonyl, 2-iodoethoxycarbonyl tert. butylcarboxyl, 4-nitrobenzyloxycarbonyl, diphenylmethoxy-carbonyl, benzhydroxycarbonyl, di-(4-methoxyphenyl-methoxycarbonyl, 2-bromoethoxycarbonyl, 2-iodoethoxycarbonyl, 2-trimethylsilylethoxycarbonyl, or 2-triphenylsilylethoxycarbonyl, etc., arylmethyoxycarbonyl having one or two aryl radicals. An additional carboxyl group in the esterified form is a silyloxycarbonyl group including an organosilyloxycarbonyl. The silicon substituents in such compounds may be substituted with lower alkyl (e.g., methyl), alkoxy (e.g., methoxy), and / or halo (e.g., chlorine). Examples of silicon substituents include trimethylsilyl and dimethyl tert. butylsilyl. In aspects of the present disclosure, the carboxyl group may be alkoxycarbonyl, particularly methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, t-butoxycarbonyl, t-pentyloxycarbonyl, cyanoheptyloxycarbonyl, particularly methoxycarbonyl or ethoxycarbonyl.

[0048] The term "imine" refers to R’1-N=CR”R’”, where R’, R”, and R’” are each independently selected from alkyl, alkenyl, carbocyclic group, heterocycle, aryl, or heteroaryl.

[0049] As used herein, the term "composition" refers to any product directly or indirectly obtained from a product containing a specified component in a specified amount and any combination of specified components in specified amounts. Such a term with respect to a pharmaceutical composition refers to a product containing the active ingredient(s) and the inert ingredient(s) constituting the carrier, as well as any product directly or indirectly resulting from the combination, complexation, or aggregation of any two or more components, or the dissociation of one or more components, or other types of reaction or interaction of one or more components. Accordingly, the pharmaceutical compositions of the present disclosure include any composition prepared by mixing the compounds of the present disclosure and a pharmaceutically acceptable carrier.

[0050] When the compounds of the present disclosure are used simultaneously with one or more other drugs, pharmaceutical compositions containing such other drugs in addition to the compounds of the present disclosure are contemplated. Accordingly, the pharmaceutical compositions of the present disclosure include those that also contain one or more other active ingredients in addition to the compounds of the present disclosure. The weight ratio of the compounds of the present disclosure to the second active ingredient may vary and will depend on the effective dosage of each component. Generally, the effective dosage of each will be used. Thus, for example, without intending to be limiting, when the compounds of the present disclosure are combined with another agent, the weight ratio of the compounds of the present disclosure to the other agent will generally be in the range of about 1000:1 to about 1:1000, preferably about 200:1 to about 1:200. The combination of the compounds of the present disclosure and other active ingredients will also generally be within the aforementioned range, but in each case, the effective dosage of each active ingredient should be used. In such combinations, the compounds of the present disclosure and the other active agents may be administered separately or together. In addition, the administration of one element may be before, simultaneous with, or after the administration of the other agent(s).

[0051] The compositions of the present disclosure can be a liquid solution, suspension, emulsion, tablet, pill, capsule, sustained release formulation, or powder. The compositions can be formulated as suppositories, along with conventional binders and carriers such as triglycerides. Oral formulations can include standard carriers such as, for example, pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like. Various delivery systems are known, and for example, encapsulation into liposomes, microparticles, and microcapsules can be used to administer the compositions of the present disclosure.

[0052] The therapeutic compositions of the present disclosure can include carriers such as one or more of polymers, carbohydrates, peptides, or derivatives thereof, which can be covalently bonded directly or indirectly to the compounds. The carriers can be substituted with substituents described herein including, but not limited to, one or more alkyl, amino, nitro, halogen, thiol, thioalkyl, sulfate, sulfonyl, sulfinyl, sulfoxide, hydroxyl groups. In an aspect of the present disclosure, the carrier is an amino acid including alanine, glycine, praline, methionine, serine, threonine, asparagine, alanyl-alanine, prolyl-methionyl, or glycyl-glycyl. The carrier can also include a molecule that targets the compounds of the present disclosure to a particular tissue or organ.

[0053] The compounds of the present disclosure can be prepared using reactions and methods that are generally known to those skilled in the art in view of the knowledge of compounds and the disclosure of this application including examples. The reactions are carried out in a solvent that is appropriate for the reagents and materials used and suitable for the reaction to be accomplished. It will be understood by those skilled in the art of organic synthesis that the functionality present in the compounds should be consistent with the proposed reaction steps. This may require modification of the order of synthetic steps or selection of a particular process scheme over another process scheme in order to obtain the desired compounds of the disclosure. It will also be recognized that another major consideration in the development of synthetic routes is the selection of protecting groups used for the protection of reactive functional groups present in the compounds described in the present disclosure. An authoritative treatise that describes many alternatives for those skilled in the art is Greene and Wuts (Protective Groups In Organic Synthesis, Wiley and Sons, 1991).

[0054] The disclosed compounds of the present disclosure can be formulated into pharmaceutical compositions for administration to a subject by suitable methods known in the art. The pharmaceutical compositions of the present disclosure, or fractions thereof, are selected based on the intended dosage form and include suitable pharmaceutically acceptable carriers, excipients, and vehicles consistent with conventional pharmaceutical practice. Suitable pharmaceutical carriers, excipients, and vehicles are described in the standard text Remington: The Science and Practice of Pharmacy (21st Edition, 2005, University of the Sciences in Philadelphia (Editor), Mack Publishing Company), and The United States Pharmacopeia: The National Formulary (USP 24 NF19) published in 1999. As an example of oral administration in the form of capsules or tablets, the active ingredient can be combined with an orally non-toxic pharmaceutically acceptable inert carrier such as lactose, starch, sucrose, methylcellulose, magnesium stearate, glucose, calcium sulfate, dicalcium phosphate, mannitol, and sorbitol. For oral administration in liquid form, the chug ingredient can be combined with any orally non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, and water. Suitable binders (such as gelatin, starch, corn sweeteners, natural sugars including glucose, natural and synthetic gums, and waxes), lubricants (such as sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride), disintegrants (such as starch, methylcellulose, agar, bentonite, and xanthan gum), flavoring agents, and coloring agents can also be combined with the compositions or their components. The compositions described herein may further include wetting agents or emulsifying agents, or pH buffering agents.

[0055] As used herein, the term "immunogenic composition" is one that, when injected into a host, elicits specific antibody production or cellular immunity.

[0056] The immunogenic compositions and / or vaccines of the present disclosure can be formulated by any of the methods known in the art. They can typically be prepared as injectable formulations, either as liquid solutions or suspensions, or as formulations for nasal administration. Prior to injection or other administration, a suitable solid form may be prepared as a solution in a liquid or a suspension in a liquid. The preparations may also, for example, be emulsified or be proteins / peptides encapsulated in liposomes.

[0057] The active immunogenic components are often mixed with excipients or carriers that are pharmaceutically acceptable and compatible with the active components. Suitable excipients include, but are not limited to, water, saline, dextrose, glycerol, or ethanol, etc., and combinations thereof. The concentration of the immunogenic polypeptide in an injectable aerosol formulation or a nasal formulation is usually in the range of about 0.2 to 5 mg / ml. Similar doses can be administered to other mucosal surfaces.

[0058] In addition, if desired, the vaccine may contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, and / or other agents that enhance the effectiveness of the vaccine. Examples of agents that may be effective include, but are not limited to, aluminum hydroxide, N-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-acetyl-nor-muramyl-L-alanyl-D-isoglutamine (CGP 11637, referred to as nor-MDP), N-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-(1'-2'-dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)-ethylamine (CGP 19835A, referred to as MTP-PE), and RIBI containing three components extracted from bacteria in a 2% squalene / Tween 80 emulsion: monophosphoryl lipid A, trehalose dimycolate, and cell wall skeleton (MPL+TDM+CWS). The effectiveness of the auxiliary substance can be determined by measuring the amount of antibody (particularly IgG, IgM, or IgA) directed against the immunogen resulting from administration of the immunogen in the vaccine containing the adjuvant in question. Additional formulations and modes of administration may also be used.

[0059] The immunogenic compositions and / or vaccines of the present disclosure can be administered in a manner compatible with dosage formulations, in a manner known in the art, and in such amounts and in such a manner as to be prophylactically and / or therapeutically effective. Generally, it is within the range of about 1 to 1,000 micrograms of viral surface envelope glycoprotein per dose and / or adjuvant molecule per dose, and more generally within the range of about 5 to 500 micrograms of glycoprotein per dose and / or adjuvant molecule per dose. The amount administered depends on the nature of the antigen and / or adjuvant molecule, the subject being treated, the ability of the host immune system to synthesize antibodies, and the degree of protection desired. The exact amount of active ingredient that needs to be administered can depend on the judgment of a physician or veterinarian and can be specific to each individual, but such determinations are within the scope of the skills of such a professional.

[0060] The vaccine or immunogenic composition can be administered as a single dose, a two-dose schedule, for example, at intervals of 2 to 8 weeks, or a multiple-dose schedule. The multiple-dose schedule can include that the primary step of vaccination can include 1 to 10 or more separate doses, followed by other doses (for example, the second dose at 1 to 4 months and, if necessary, subsequent doses (plural) several months later) administered at subsequent time intervals necessary to maintain and / or enhance the immune response.

[0061] As used herein, the term "immunogenic fragment" refers to a fragment of an immunogen that contains one or more epitopes and thus can modulate an immune response or act as an adjuvant to an antigen administered concomitantly. Such fragments can be identified using any number of epitope mapping techniques well known in the art (see, for example, Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66 (Morris, G.E., Ed., 1996) Humana Press, Totowa, NJ).

[0062] The immunogenic fragment can be at least about 2 amino acids in length, more preferably about 5 amino acids in length, and most preferably at least about 10 to about 15 amino acids in length. There is no critical upper limit to the length of the fragment, which can include substantially the full-length of the protein sequence or even a fusion protein containing two or more epitopes.

[0063] As used herein, the term "immunoglobulin" refers to a class of proteins that exhibit antibody activity and bind to other molecules (e.g., antigens and certain cell surface receptors) with a high degree of specificity. Immunoglobulins can be classified into five classes: IgM, IgG, IgA, IgD, and IgE. IgG is the most abundant class of antibodies in the body and is envisioned to have a twisted "Y" shape structure. Except for IgM, immunoglobulins are composed of four peptide chains linked by intra-chain and inter-chain disulfide bonds. IgG is composed of two polypeptide heavy chains (H chains) and two polypeptide light chains (L chains) coupled by non-covalent disulfide bonds.

[0064] As used herein, the term "immunological response" refers to the development of a humoral and / or cellular immune response in a subject to an antigen present in a composition of interest. For the purposes of the present disclosure, a "humoral immune response" refers to an immune response mediated by antibody molecules, and a "cellular immune response" is an immune response mediated by T lymphocytes and / or other white blood cells.

[0065] One aspect of cellular immunity involves an antigen-specific response by cytotoxic T cells (“CTLs”). CTLs associate with and are presented by proteins encoded by the major histocompatibility complex (MHC) and have specificity for peptide antigens expressed on the surface of cells. CTLs serve to induce and facilitate the destruction of intracellular microorganisms or the lysis of cells infected with such microorganisms. Another aspect of cellular immunity involves an antigen-specific response by helper T cells. Helper T cells act to stimulate the functions of non-specific effector cells and to focus the activity of non-specific effector cells on cells that associate with MHC molecules on their surface and present peptide antigens. “Cellular immune response” also refers to the production of cytokines, chemokines, and other such molecules produced by activated T cells and / or other leukocyte cells, including those derived from CD4+ and CD8+ T cells. Thus, an immunological response can include one or more of the following effects: production of antibodies by B cells and / or activation of suppressor T cells and / or γδ T cells specifically directed against one or more antigens present in the composition or vaccine of interest. These responses can function to neutralize infectivity and / or provide protection to the immunized host by mediating antibody complement or antibody-dependent cell cytotoxicity (ADCC). Such responses can be determined using standard immunoassays and neutralization assays well known in the art.

[0066] As used herein, the term “immunogenic amount” refers to an amount capable of inducing the production of antibodies directed against a virus in a host to which the vaccine has been administered.

[0067] As used herein, the term "immunogenic carrier" refers to a composition that enhances the immunogenicity of virosomes from any of the viruses discussed herein. Such carriers include, but are not limited to, proteins and polysaccharides, and microspheres formulated using biodegradable polymers such as, for example, DL-lactide-coglycolide, liposomes, and bacterial cells and membranes. Protein carriers can be conjugated to proteases or peptides derived therefrom to form fusion proteins by recombinant or synthetic techniques or by chemical coupling. Useful carriers and methods of coupling such carriers to polypeptide antigens are known in the art.

[0068] As used herein, the term "immunopotentiator" is intended to mean a substance that, when mixed with an immunogen, elicits a higher immune response than the immunogen alone. For example, an immunopotentiator can enhance immunogenicity and provide an excellent immune response. An immunopotentiator can act, for example, by enhancing the expression of co-stimulators on macrophages and other antigen-presenting cells.

[0069] As used herein, the terms "subject", "individual", or "patient" are used interchangeably and refer to animals, preferably warm-blooded animals such as mammals. Mammals include, but are not limited to, any member of the class Mammalia. Mammals as subjects or patients in the present disclosure can be from the families of primates, carnivores, proboscideans, perissodactyls, artiodactyls, rodents, and lagomorphs. In certain embodiments, the mammal is a human. In other embodiments, animals can be treated, and the animals can be vertebrates including both birds and mammals. In aspects of the present disclosure, the term includes food or pet livestock including horses, cows, sheep, poultry, fish, pigs, dogs, cats, and zoo animals, goats, apes (e.g., gorillas or chimpanzees), and rodents such as rats and mice.

[0070] As used herein, the term "pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient, or vehicle for use in animals and more particularly in humans, which is approved by a regulatory agency of the Federal or State government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeias, and with which the disclosed probes are administered together. Such pharmaceutical carriers can be liquids such as, for example, water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. The pharmaceutical carrier can be, for example, physiological saline solution, acacia gum, gelatin, starch paste, talc, keratin, colloidal silica, and urea. When administered to a patient, the probe and the pharmaceutically acceptable carrier can be sterilized. When the probe is administered intravenously, water is a useful carrier. Aqueous saline solutions, as well as aqueous dextrose and glycerol solutions, can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical carriers can also include excipients such as, for example, glucose, lactose, sucrose, glycerol monostearate, sodium chloride, glycerol, propylene glycol, water, and ethanol. Optionally, the composition can also contain small amounts of wetting or emulsifying agents, or pH buffering agents. The composition can advantageously be in the form of a solution, an emulsion, a sustained release formulation, or any other form suitable for use.

[0071] As used herein, the term "pharmaceutically acceptable" is used to refer to compounds, materials, compositions, and / or dosage forms that are suitable for use in contact with the tissues of humans and animals within the scope of sound medical judgment, without undue toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0072] As used herein, the term "vaccine" refers to an immunogenic amount of one or more virosomes, fragments thereof, or subunits thereof. Such vaccines can include them in combination with one or more additional viral components, such as other proteins or other immunogens, that naturally associate with one or more viral surface envelope glycoproteins and portions thereof, as well as adjuvant molecules and portions thereof on the surface of the virosomes, or viral particles or epitope peptides derived therefrom.

[0073] As used herein, the terms "effective amount" and "an effective amount" refer to an amount that achieves a desired result, is effective in preventing an undesirable condition or disease or condition, or enhances and / or modulates the immune system of a subject to a desired response against a particular pathogen (e.g., a virus), or is sufficient to prevent a disease or condition. For example, a "therapeutically effective amount" is an amount sufficient to achieve a desired therapeutic result, is effective against an undesirable symptom, or is sufficient to prevent a disease or symptom and / or modulate the immune system of a subject to a desired response against a particular pathogen, but generally an amount insufficient to cause harmful side effects. Any specific therapeutically effective dosage level for a particular patient will depend on a variety of factors including the specific composition used; the age, weight, general health, sex, and diet of the patient; the time of administration; the route of administration; the rate of excretion of the specific compound used; the duration of the treatment; drugs used in combination with or concurrently with the specific compound used, and like factors well known in the medical arts. For example, it is well within the skill of the art to start the dosage of a compound at a level lower than that required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. If desired, an effective daily dosage can be divided into multiple dosages for administration. Consequently, a single-dose composition can contain such an amount or an approximation thereof to constitute a daily dosage. In the event of any contraindications, the dosage can be adjusted by the individual physician. Dosages can vary and can be administered in one or more dosages per day for one or more days. Guidance can be found in the literature for appropriate dosages for a given class of pharmaceutical products.

[0074] As used herein, the terms "treating" and "treatment" can generally refer to obtaining a desired pharmacological and / or physiological effect. The effect may be prophylactic with respect to preventing or partially preventing a disease, symptom, or condition, such as an infection and its consequences, and / or modulating the subject's immune system for a desired response against a particular pathogen, but need not necessarily be so. The effect can be therapeutic in terms of partial or complete cure of a disease, condition, symptom, or adverse effect resulting from a disease, disorder, or condition. The term "treatment" as used herein can include any treatment of an infectious disease in a subject, particularly a human, and can include any one or more of the following: (a) preventing the development of a disease in a subject who has a predisposition to the disease or infection but has not yet been diagnosed as having the disease, (b) suppressing a disease or infection, i.e., preventing its onset, and (c) alleviating a disease or infection, i.e., alleviating or improving the disease and / or its symptoms or condition, and (d) and / or modulating the subject's immune system for a desired response against a particular pathogen. The term "treatment" as used herein can refer to either therapeutic treatment alone, prophylactic treatment alone, or both therapeutic treatment and prophylactic treatment. Those in need of treatment (subjects in need of treatment) can include those who already have a disorder and / or those who should prevent a disorder. The term "treating" as used herein can include suppressing a disease, disorder, or condition, e.g., preventing its progression, and alleviating a disease, disorder, or condition, e.g., causing regression of the disease, disorder, and / or condition. And / or modulating the subject's immune system for a desired response against a particular pathogen. Treating a disease, disorder, or condition can include improving at least one symptom of a particular disease, disorder, or condition, even if the underlying pathophysiology is not affected, e.g., treating a subject's pain by administration of an analgesic even though such an agent does not treat the cause of the pain.

[0075] As used herein, "treatment" can refer to the treatment, cure, and / or amelioration of a disease, disorder, condition, or side effect, or the reduction in the rate of progression of a disease, disorder, condition, or side effect, and / or the modulation of the immune system of a subject to a desired response to a particular pathogen.

[0076] The term "pharmaceutically acceptable" describes a material that is not biologically or otherwise undesirable, i.e., a material that does not cause unacceptable levels of undesirable biological effects or interact in a harmful manner.

[0077] The term "pharmaceutically acceptable prodrug" or "prodrug" refers to prodrugs of the compounds of the present disclosure that are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., within the scope of sound medical judgment, commensurate with a reasonable benefit / risk ratio, and effective for their intended use. The prodrugs of the present disclosure can be rapidly converted in vivo, for example, by hydrolysis in the blood, to the parent compound having the structure of the disclosed compound. Detailed discussions are provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, V. 14 of the A.C.S. Symposium Series, and Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press (1987).

[0078] In 1974, Dalsgaard et al. ("Saponin adjuvants", Archiv. fur die gesamte Virusforschung, Vol. 44, Springer Verlag, Berlin, p243-254) first described that a saponin preparation isolated from the South American tree Quillaja saponaria Molina has adjuvant activity. The purified fragment of Quil A has been isolated by HPLC, and this fragment retains adjuvant activity without the toxicity associated with Quil A (EP 0 362 278), for example, QS7 and QS21 (also known as QA7 and QA21). QS-21 is a natural saponin derived from the bark of Quillaja sapanaria Molina that induces CD8+ cytotoxic T cells (CTLs), Th1 cells, and a predominant IgG2a antibody response.

[0079] The saponins of the present disclosure can be used at a level of 1 to 100 μg, about 50 μg, for example 40 to 60 μg, preferably 45 to 55 μg, or 49 to 51 μg, or 50 μg per human dose of the adjuvant composition. In some embodiments, the human dose of the adjuvant composition can contain QS21 at a level of about 25 μg, for example 20 to 30 μg, preferably 21 to 29 μg, or 22 to 28 μg, or 28 to 27 μg, or 24 to 26 μg, or 25 μg.

[0080] When an adjuvant is combined with the liquid form of an antigenic composition, the adjuvant composition is a suitable volume for a human dose, which is approximately half of the intended final dose of the human dose. For example, for an intended final human dose of 1 μl, the volume of the adjuvant is 500 μl, or for an intended final human dose of 0.5 ml, the volume is 250 μl. When combined with the antigen composition, the adjuvant composition is diluted to provide a vaccine of the final human dose. The final volume of such a dose will of course vary depending on the initial volume of the adjuvant composition and the volume of the antigen composition added to the adjuvant composition. In an alternative embodiment, the aqueous adjuvant is used to reconstitute the lyophilized antigen composition. In this embodiment, the volume suitable for a human dose of the adjuvant composition is approximately equal to the final volume of the human dose. The liquid adjuvant composition is added to a vial containing the lyophilized antigen composition and is used to reconstitute the lyophilized antigen composition.

[0081] Abbreviations IgG, immunoglobulin G; Th, T helper cell; CTL, cytotoxic T lymphocyte; rha, rhamnose; xyl, xylose; OVA, ovalbumin; NMM, N-methylmorpholine; HOBt, hydroxybenzotriazole; EDC HCl, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; DCM, dichloromethane; MeCN, acetonitrile; THF, tetrahydrofuran; rHagB, recombinant hemagglutinin B; s.c., subcutaneous; ESI-TOF, electrospray ionization time-of-flight mass spectrometer; ELISA, enzyme-linked immunosorbent assay;

[0082] Discussion The present disclosure provides compounds containing modified saponin compounds, pharmaceutical compositions containing modified saponin compounds, methods of using modified saponin compounds and pharmaceutical compositions, and methods of making modified saponin compounds. The compounds and pharmaceutical compositions of the present disclosure can be used in combination with one or more other therapeutic agents for treating viral infections and other diseases. For example, the compounds and pharmaceutical compositions of the present disclosure can be used in combination with other antiviral agents for treating viral infections.

[0083] One aspect of the present disclosure encompasses embodiments of modified saponins having the following formula:

[0084] [Chemical formula]

[0085] (wherein, q1 can be H or OH, q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a constituent of an acetal group, or an imine group, f3 and f4 are each independently OH, or acetyl, or C3 and C4 of a fuocsyl unit, and f3 and f4 can form a cyclic ketal ring or a cyclic carbonate ester, f5 is selected from H, a methyl group, -CH2OH group, R4-NR5-C(O)-, R4-O-C(O)-, or R4-O-CH2-, R4 and R5 are each independently a straight chain having the structure R6[(CX2) 0-20 O 0-1 (CY2) 0-20 0-20 and X and Y are each independently H or a halogen atom, R6 is COOR7, C(O)NR7R8, NR7R8, or OR7, R7 is Ar[(CZ2) 0-20 O 0-1 (CL2) 0-20 0-20 and R8 is H or alkyl,​​ Z and L are each independently H or a halogen atom, Ar is a substituted or unsubstituted aromatic system, ga5 is R 14 -NR 15 -C(O)-, R 14 -O-C(O)-, or R 14 -O-CH2-, and is selected from R 14 and R 15 are each independently a straight chain having the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 0-20 and X1 and Y1 are each independently H or a halogen atom, R 16 is COOR 17 C(O)NR 17 R 18 NR 17 R 18 or OR 17 and R 17 is Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 0-20 and R 18 is H or alkyl, Z1 and L1 are each independently H or a halogen atom, Ar2 is a substituted or unsubstituted aromatic group, r3 is H, a monosaccharide, a disaccharide, or a trisaccharide, x3 is H, a monosaccharide (excluding xylose), or a disaccharide, and ga3 is H, a monosaccharide, or a disaccharide).

[0086] In some embodiments, the modified saponin may have the following formula I:

[0087]

Chemical formula

[0088] (In the formula, q1 is H or OH; q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a component of an acetal group, or an imine group; ga5 is R 14 -NR 15 -C(O)-, R 14 -OC(O)- or R 14 -O-CH2-, R 14 and R 15 each independently represents a group of the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 is a straight chain having X1 and Y1 are each independently H or a halogen atom; R 16 But COOR 17 , C(O)NR 17 R 18 , N.R. 17 R 18 OR 17 and R 17 But Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 and R 18 is H or alkyl, Z1 and L1 are each independently H or a halogen atom; and Ar2 is a substituted or unsubstituted aromatic group.

[0089] In other embodiments, the modified saponin may be selected from the group consisting of formulas 2D-2H below.

[0090] [ka]

[0091] Another aspect of the present disclosure encompasses embodiments of a pharmaceutical composition comprising a modified saponin having the following formula:

[0092] [Chemical formula]

[0093] (wherein, q1 can be H or OH, q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a constituent of an acetal group, or an imine group, f3 and f4 are each independently OH, or acetyl, or C3 and C4 of a fuocsyl unit, and f3 and f4 can form a cyclic ketal ring or a cyclic carbonate ester, f5 is selected from H, a methyl group, -CH2OH group, R4-NR5-C(O)-, R4-O-C(O)-, or R4-O-CH2-, R4 and R5 are each independently a straight chain having the structure R6[(CX2) 0-20 O 0-1 (CY2) 0-20 0-20 and X and Y are each independently H or a halogen atom, R6 is COOR7, C(O)NR7R8, NR7R8, or OR7, R7 is Ar[(CZ2) 0-20 O 0-1 (CL2) 0-20 0-20 and R8 is H or alkyl, Z and L are each independently H or a halogen atom, Ar is a substituted or unsubstituted aromatic system, ga5 is selected from R 14 -NR 15 -C(O)-, R 14 -O-C(O)-, or R 14 -O-CH2-, R 14 and R 15 are each independently a structure R 16 ​​[(CX12) 0-20 O 0-1 (CY12) 0-20 0-20 is a straight chain having X1 and Y1 are each independently H or a halogen atom, R 16 is COOR 17 , C(O)NR 17 R 18 , NR 17 R 18 , or OR 17 and R 17 is Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 0-20 and R 18 is H or alkyl, Z1 and L1 are each independently H or a halogen atom, Ar2 is a substituted or unsubstituted aromatic group, r3 is H, a monosaccharide, a disaccharide, or a trisaccharide, x3 is H, a monosaccharide (excluding xylose), or a disaccharide, and ga3 is H, a monosaccharide, or a disaccharide).

[0094] Another aspect of the present disclosure encompasses embodiments of a pharmaceutical composition comprising a modified saponin having the following formula I:

[0095] [Chemical formula]

[0096] (wherein q1 is H or OH, q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a constituent of an acetal group, or an imine group, ga5 is R 14 -NR 15 -C(O)-, R 14 -O-C(O)-, or R​​14 selected from -O-CH2- R 14 and R 15 are each independently a straight chain having the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 0-20 wherein X1 and Y1 are each independently H or a halogen atom, and R R 16 is COOR 17 , C(O)NR 17 R 18 , NR 17 R 18 , or OR 17 ; and R 17 is Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 0-20 where R 18 is H or alkyl, Z1 and L1 are each independently H or a halogen atom, and Ar2 is a substituted or unsubstituted aromatic group).

[0097] Another aspect of the present disclosure encompasses embodiments of a pharmaceutical composition comprising a modified saponin having the following formulas 2D - 2H.

[0098]

Chemical formula

[0099] ​​In some embodiments of this aspect of the present disclosure, the pharmaceutical composition comprises a therapeutically effective amount of a modified saponin for treating a condition (e.g., an infectious disease, a disorder, etc.) of a subject (e.g., an animal or a human subject). The pharmaceutical composition may further comprise at least one immunogen (e.g., an immunogenic amount), an agent (e.g., an anti-cancer agent), and a pharmaceutically acceptable carrier, etc. The present disclosure provides administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a modified saponin or a pharmaceutically acceptable salt of an active agent modified saponin, and a pharmaceutically acceptable carrier.

[0100] The condition to be treated of a subject (e.g., a mammal) in need of treatment can include the condition that the vaccine directly targets. This condition can be an infection (e.g., a pathogen infection such as a viral infection such as coronavirus infection, HIV infection, influenza infection, hepatitis), or a disorder such as cancer, an infectious or non-viral excessive inflammation / immune hyperactivity state.

[0101] In some embodiments of this aspect of the present disclosure, the pharmaceutical composition may further comprise at least one anti-cancer agent. At least one chemotherapeutic agent and a saponin derivative may be mixed in a pharmaceutically acceptable formulation, or may be covalently bonded to each other together with a pharmaceutically acceptable carrier. The anti-cancer agent can be present in a therapeutically effective amount to treat the condition.

[0102] Yet another aspect of the present disclosure encompasses embodiments of a method for increasing the immunogenicity of an immunogen when administered to an animal or a human subject, the method comprising at least the step of administering to the subject a vaccine comprising a pharmaceutical composition according to the present disclosure.

[0103] Yet another aspect of the present disclosure encompasses embodiments of a synthetic route for synthesizing a saponin derivative, the synthetic route comprising coupling a natural saponin with a functionalized side chain molecule, the functionalized side chain comprising an amino group or a hydroxyl group.

[0104] In some embodiments of this aspect of the present disclosure, the natural saponin can be obtained from Momordica cochinchinensis Spreng.

[0105] In some embodiments of this aspect of the disclosure, the natural saponin can be coupled to a functionalized side chain molecule via an amide formation reaction or an ester formation reaction.

[0106] Pharmaceutical Formulations and Routes of Administration Embodiments of the present disclosure include the agents (e.g., modified saponins) identified herein and can be formulated with one or more pharmaceutically acceptable excipients, diluents, carriers, and / or adjuvants. In addition, embodiments of the present disclosure include agents formulated with one or more pharmaceutically acceptable auxiliary substances. In particular, the agent can be formulated with one or more pharmaceutically acceptable excipients, diluents, carriers, and / or adjuvants to provide embodiments of the compositions of the present disclosure.

[0107] A wide variety of pharmaceutically acceptable excipients are known in the art. Pharmaceutically acceptable excipients are described in detail in various publications, including, for example, A. Gennaro (2000) "Remington: The Science and Practice of Pharmacy" 20th edition, Lippincott, Williams, & Wilkins, Pharmaceutical Dosage Forms and Drug Delivery Systems (1999) H.C. Ansel et al., eds., 7 th ed., Lippincott, Williams, & Wilkins, and Handbook of Pharmaceutical Excipients (2000) A.H. Kibbe et al., eds., 3 rd ed. Amer. Pharmaceutical Assoc.

[0108] Pharmaceutically acceptable excipients such as vehicles, adjuvants, carriers, or diluents are generally readily available. Furthermore, pharmaceutically acceptable auxiliary substances such as, for example, pH adjusters and buffers, tonicity adjusters, stabilizers, and wetting agents are generally readily available.

[0109] In one embodiment of the present disclosure, the agent can be administered to a subject using any means capable of producing the desired effect. Thus, the agent can be incorporated into various formulations for therapeutic administration. For example, the agent can be formulated into a pharmaceutical composition by combination with a suitable pharmaceutically acceptable carrier or diluent, and can be formulated into preparations in solid, semi-solid, liquid, or gaseous forms such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, and aerosols.

[0110] In pharmaceutical formulations, the agent may be administered in the form of its pharmaceutically acceptable salts, or the subject active composition may be used alone, or in suitable combinations, and further in combination with other pharmaceutically active compounds. The following methods and excipients are merely illustrative and in no way limiting.

[0111] In the case of oral preparations, the agent can be used alone or in combination with suitable additives, for example, conventional additives such as lactose, mannitol, corn starch, potato starch, and binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch, or gelatin, and disintegrants such as corn starch, potato starch, or sodium carboxymethyl cellulose, and lubricants such as talc or magnesium stearate, and, if desired, diluents, buffers, wetting agents, preservatives, and flavorings, in combination to produce tablets, powders, granules, or capsules.

[0112] Embodiments of the agent can be formulated into injectable preparations, if desired, by dissolving, suspending, or emulsifying them in vegetable oils or other similar oils, synthetic fatty acid glycerides, higher fatty acid esters, or aqueous or non-aqueous solvents such as propylene glycol, together with conventional additives such as solubilizing agents, isotonic agents, suspending agents, emulsifying agents, stabilizers, and preservatives.

[0113] Embodiments of the agent can be utilized in aerosol formulations administered via inhalation. Embodiments of the agent can be formulated, for example, with pressurized acceptable propellants such as dichlorodifluoromethane, propane, and nitrogen.

[0114] Furthermore, embodiments of the agent can be made into suppositories by mixing with various bases such as emulsifying bases or water-soluble bases. Embodiments of the agent can be administered rectally via suppositories. Suppositories can contain vehicles such as cocoa butter, cacao wax, polyethylene glycol, etc., which melt at body temperature but solidify at room temperature.

[0115] Unit dosage forms for oral or rectal administration such as syrups, elixirs, and suspensions can be provided, and each dosage unit, for example, one cup of teaspoon, one cup of tablespoon, tablets, or suppositories, contains a predetermined amount of the composition containing one or more compositions. Similarly, unit dosage forms for injection or intravenous administration can contain the agent in the composition as a solution in sterile water, physiological saline, or another pharmaceutically acceptable carrier.

[0116] Embodiments of the agent can be formulated into injectable compositions according to the present disclosure. Usually, injectable compositions can be prepared as liquid solutions or suspensions, and prior to injection, suitable solid forms can also be prepared as solutions or suspensions in liquid vehicles. The preparations can also be emulsified according to the present disclosure, or the active ingredient (triaminopyridine derivative and / or labeled triaminopyridine derivative) can be encapsulated in liposomal vehicles.

[0117] In an embodiment, the agent can be formulated for delivery by a continuous delivery system. The term "continuous delivery system" is used interchangeably herein with "controlled delivery system" and includes continuous (e.g., controlled) delivery devices (e.g., pumps) combined with catheters, injection devices, etc., and various types thereof are known in the art.

[0118] Mechanical or electrically powered infusion pumps may also be suitable for use in the present disclosure. Examples of such devices include, for example, those described in U.S. Patent Nos. 4,692,147, 4,360,019, 4,487,603, 4,360,019, 4,725,852, 5,820,589, 5,643,207, 6,198,966, etc. Generally, delivery of the agent can be achieved using any of a variety of refillable pump systems. The pump provides consistent, controlled release over a long period of time. In some embodiments, the agent can be present in a liquid formulation within a drug-impermeable reservoir and delivered continuously to the subject.

[0119] In one embodiment, the drug delivery system is at least partially an implantable device. The implantable device can be implanted at any suitable implantation site using methods and devices well known in the art. The implantation site is a site within the body of the subject into which the drug delivery device is introduced and placed. The implantation site includes, but is not necessarily limited to, subdermal, subcutaneous, intramuscular, or other suitable sites within the body of the subject. Subcutaneous implantation sites are used in some embodiments because they are easy to implant and remove the drug delivery device.

[0120] Drug delivery devices suitable for use in the present disclosure may be based on any of a variety of modes of operation. For example, the drug delivery device may be based on a diffusion system, a convection system, or an erosion system (e.g., an erosion-based system). For example, the drug delivery device can be an electrochemical pump, an osmotic pump, an electroosmotic pump, a vapor pressure pump, or an osmotic burst matrix. For example, the drug is incorporated into a polymer, and the polymer provides release of the pharmaceutical formulation simultaneously with the degradation of the drug-impregnated polymer material (e.g., a biodegradable drug-impregnated polymer material). In other embodiments, the drug delivery device is based on an electro-diffusion system, an electrolytic pump, a foam pump, a piezoelectric pump, a hydrolysis system, and the like.

[0121] Drug delivery devices based on mechanical or electromechanical infusion pumps may also be suitable for use in the present disclosure. Examples of such devices include, for example, U.S. Patent Nos. 4,692,147, 4,360,019, 4,487,603, 4,360,019, and 4,725,852, among others. In general, the subject treatment methods can be achieved using any of a variety of refillable and non-replaceable pump systems. Pumps and other convective systems are generally preferred because they generally provide more consistent, long-term controlled release. Osmotic pumps are used in some embodiments due to the combined advantages of more consistent controlled release and relatively small size (see, for example, PCT Publication No. WO97 / 27840 and U.S. Patent Nos. 5,985,305 and 5,728,396). Examples of osmotic drive devices suitable for use in the present disclosure include, but are not necessarily limited to, U.S. Patent Nos. 3,760,984, 3,845,770, 3,916,899, 3,923,426, 3,987,790, 3,995,631, 3,916,899, 4,016,880, 4,036,228, 4,111,202, 4,111,203, 4,203,440, 4,203,442, 4,210,139, 4,327,725, 4,627,850, 4,865,845, 5,057,318, 5,059,423, 5,112,614, 5,137,727, 5,234,692, 5,234,693, 5,728,396, and the like.

[0122] In some embodiments, the drug delivery device is an implantable device. The drug delivery device can be implanted at any suitable implantation site using methods and devices well known in the art. As described herein, an implantation site is a site within the body of a subject into which the drug delivery device is introduced and placed. Implantation sites include, but are not necessarily limited to, subdermal, subcutaneous, intramuscular, or other suitable sites within the body of the subject.

[0123] In some embodiments, the agent can be delivered using an implantable drug delivery system, such as a programmable system for administering the agent. Exemplary programmable implantable systems include implantable infusion pumps. Exemplary implantable infusion pumps, or devices useful in connection with such pumps, are described, for example, in U.S. Patent Nos. 4,350,155; 5,443,450; 5,814,019; 5,976,109; 6,017,328; 6,171,276; 6,241,704; 6,464,687; 6,475,180; and 6,512,954. A further exemplary device that can be adapted to the present disclosure is the SynchroMed infusion pump (Medtronic).

[0124] Excipient vehicles suitable for the agent are, for example, water, saline, dextrose, glycerol, ethanol, etc., and combinations thereof. Additionally, if desired, the vehicle may contain trace amounts of auxiliary substances such as wetting agents, emulsifying agents, or pH buffering agents. Methods for preparing such dosage forms are known to those of skill in the art or will be apparent in view of the present disclosure. See, for example, Remington’s Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania, 17th edition, 1985. In any event, the composition or formulation to be administered will contain an amount of the agent sufficient to achieve the desired condition in the subject being treated.

[0125] The compositions of the present disclosure can include those that contain a sustained-release or controlled-release matrix. Additionally, embodiments of the present disclosure can be used in combination with other treatments that use sustained-release formulations. As used herein, a sustained-release matrix is a matrix made of a material, usually a polymer, that is degradable by enzymatic hydrolysis or acid-base hydrolysis, or by dissolution. When inserted into the body, enzymes and body fluids act on this matrix. The sustained-release matrix is preferably selected from biocompatible materials such as liposomes, polylactide (polylactic acid), polyglycolide (a polymer of glycolic acid), polylactide-co-glycolide (a copolymer of lactic acid and glycolic acid), polyanhydrides, poly(ortho)esters, polypeptides, hyaluronic acid, collagen, chondroitin sulfate, carboxylic acids, fatty acids, phospholipids, polysaccharides, nucleic acids, polyamino acids, amino acids such as phenylalanine, tyrosine, isoleucine, polynucleotides, polyvinylpropylene, polyvinylpyrrolidone, and silicones. Exemplary biodegradable matrices include a polylactic acid matrix, a polyglycolic acid matrix, and a polylactic acid-glycolic acid (lactic acid-glycolic acid copolymer) matrix.

[0126] In another embodiment, the pharmaceutical compositions (and combination compositions) of the present disclosure can be delivered by a controlled release system. For example, the agent can be administered using intravenous infusion, an implantable osmotic pump, a transdermal patch, liposomes, or other modes of administration. In one embodiment, a pump (Sefton (1987). CRC Crit. Ref. Biomed. Eng. 14:201, Buchwald et al. (1980). Surgery 88:507, Saudek et al. (1989). N. Engl. J. Med. 321:574) can be used. In another embodiment, a polymeric material is used. In yet another embodiment, the controlled release system is placed near the therapeutic target and thus requires only a fraction of the systemic dose. In yet another embodiment, the controlled release system is placed near the therapeutic target and thus requires only a fraction of the body. Other controlled release systems are discussed in a review by Langer (1990). Science 249:1527-1533.

[0127] In another embodiment, the compositions of the present disclosure (and combination compositions individually or together) include those formed by impregnating the agents described herein into an absorbent material, such as sutures, bandages, and gauze, or coating the surface of a solid phase material such as surgical staples, zippers, and catheters, for delivering the composition. Other delivery systems of this type will be readily apparent to those skilled in the art in view of the present disclosure.

[0128] Dosage Embodiments of the agent (e.g., modified saponin) can be administered to a subject in one or more doses. One skilled in the art will readily appreciate that the dosage level can vary depending on the function of the particular agent being administered, the severity of the condition, and the subject's susceptibility to side effects. The preferred dosage of a given compound can be readily determined by one skilled in the art by various means.

[0129] In the embodiment, the drug is administered multiple times. The dosing frequency of the drug can vary depending on various factors, such as the severity of the symptoms, for example. For example, in the embodiment, the drug can be administered once a month, twice a month, three times a month, every other week (qow), once a week (qw), twice a week (biw), three times a week (tiw), four times a week, five times a week, six times a week, every other day (qod), daily (qd), twice a day (qid), or three times a day (tid). As discussed above, in the embodiment, the drug is administered continuously.

[0130] The administration period of the drug, for example, the period during which the drug is administered, can vary depending on any of various factors, such as the patient's response, for example. For example, the drug can be administered in combination or individually for a period of about 1 day to 1 week, about 2 weeks to 4 weeks, about 1 month to 2 months, about 2 months to 4 months, about 4 months to 6 months, about 6 months to 8 months, about 8 months to 1 year, about 1 year to 2 years, or about 2 years to 4 years, or longer.

[0131] A dosage at a high concentration of 60 micrograms / kilogram is non-toxic. Also, even at lower concentrations, such as 1 to 4 micrograms / kilogram, it exhibits biological activity in an in vivo system. 10 established in vitro -9 ~10 -6The concentration of M is active and this concentration is expected to be achieved even in the cellular environment (Slominski AT, Janjetovic Z, Fuller BE, Zmijewski MA, Tuckey RC, et al. (2010) Products of vitamin D3 or 7-dehydrocholesterol metabolism by cytochrome P450scc show anti-leukemia effects, having low or absent calcemic activity. PLoS ONE 5(3):e990; Slominski AT, Kim T-K., Janjetovic Z, Tuckey RC, Bieniek, R, Yue Y, Li W, Chen J, Miller D, Chen T, Holick M(2011) 20-hydroxyvitamin D2 is a non-calcemic analog of vitamin D with potent antiproliferative and prodifferentiation activities in normal and malignant cells. Am J Physiol:Cell Physiol 300:C526-C541; Wang J, Slominski AT, Tuckey RC, Janjetovic Z, Kulkarni A, Chen J, Postlethwaite A, Miller D, Li W(2012) 20-Hydroxylvitamin D3possesses high efficacy against proliferation of cancer cells while being non-toxic.See Anticancer Res 32:739-746; Slominski A, Janjetovic Z, Tuckey RC, Nguyen MN, Bhattacharya KG, Wang J, Li W, Jiao Y, Gu W, Brown M, Postlethwaite AE (2013) 20-hydroxyvitamin D3, noncalcemic product of CYP11A1 action on vitamin D3, exhibits potent antifibrogenic activity in vivo. J Clin Endocrinol Metab 98, E298-E30; Chen, J, J. Wang, T. Kim, E. Tieu, E. Tamg, Lin Z, D. Kovacic, D. Miller, A. Postlethwaite, R. Tuckey, A. Slominski and W. Li (2014). Novel Vitamin D Analogs as Potential Therapeutics: The Metabolism, Toxicity Profiling, and Antiproliferative Activity. Anticancer Res 34:2153-2163).

[0132] In one embodiment, the dosage for administering any single agent of the present disclosure to a subject (e.g., a mammal such as a human) having a condition (e.g., COVID-19) is about 2 to 60 micrograms / kilogram, or in the case of a combination of agents, each agent can be about 2 to 60 micrograms / kilogram.

[0133] Route of administration Embodiments of the present disclosure provide methods and compositions for administering an agent (e.g., a modified saponin) to a subject (e.g., a human) using any available method and route suitable for drug delivery (including in vivo and in vitro methods, as well as systemic and local administration routes).

[0134] The administration routes include intranasal, intramuscular, intratracheal, subcutaneous, intradermal, topical application, intravenous, rectal, transnasal, oral, and other enteral and parenteral administration routes. The administration routes can be combined if desired or adjusted according to the drug and / or the desired effect. The drug can be administered as a single dose or multiple doses.

[0135] Embodiments of the drug can be administered to a subject using conventional methods and routes available for the delivery of conventional drugs, including systemic or local routes. Generally, the administration routes contemplated by the present disclosure include, but are not limited to, enteral routes, parenteral routes, or inhalation routes.

[0136] The parenteral administration route includes, as routes other than inhalation administration, but is not limited to, local, transdermal, subcutaneous, intramuscular, intraorbital, intraarticular, intraspinal, intrathoracic, and intravenous routes, that is, any administration route other than those through the digestive tract. Parenteral administration can be carried out to achieve systemic or local delivery of the drug. If systemic delivery is desired, the administration typically involves invasive or systemic absorption of a pharmaceutical preparation by local or mucosal administration.

[0137] In embodiments, the drug can also be delivered to the subject by enteral administration. The enteral administration route includes, but is not limited to, oral and rectal (e.g., using suppositories) delivery.

[0138] Methods for administering the drug through the skin or mucosa include, but are not limited to, topical application of a suitable pharmaceutical preparation, transdermal delivery, injection, and epidermal administration. For transdermal delivery, penetration enhancers or iontophoresis are suitable methods. Iontophoresis can be performed using commercially available "patches" that continuously deliver the product through intact skin via electrical pulses over a period of several days or more.

[0139] Embodiments of the present disclosure are described in connection with examples and corresponding text and drawings, but the present disclosure is not intended to be limited to these described embodiments. On the contrary, the intention is to cover all alternatives, modifications, and equivalents included within the spirit and scope of the embodiments of the present disclosure.

[0140] Embodiments of the present disclosure are described in connection with examples and corresponding text and drawings, but the present disclosure is not intended to be limited to these described embodiments. On the contrary, the intention is to cover all alternatives, modifications, and equivalents included within the spirit and scope of the embodiments of the present disclosure.

[0141] Example 1

[0142] [Chemical Formula]

[0143] Synthesis of 2. (BOC)2O (33 mg, 0.15 mmol) was added to 11-amino-1-undecanol (20 mg, 0.1 mmol) which was being stirred at 0 °C in N,N-dimethylformamide (0.5 mL) and NaHCO3 (5 mg, 20%, v / v). The reaction mixture was stirred at room temperature overnight. Then, the reaction mixture was filtered by column chromatography packed with silica gel and ethyl acetate to obtain 2, which was used directly for the synthesis of 3a and 3b.

[0144] Synthesis of 3a. Benzoyl chloride (230 μl, 2 mmol) was added to intermediate 2 ((30 mg, 0.1 mmol) while stirring in a mixture of dichloromethane (2.25 mL) and triethylamine (1.39 mL, 10 mmol) at room temperature. The mixture was stirred for 48 hours and then concentrated on a rotary evaporator and extracted with EtOAc. The organic layer was washed three times with NaHCO3 and dried over Na2SO4. Then, the collected organic layer was purified by silica gel column chromatography (eluting with a gradient of petroleum ether and ethyl acetate) to obtain 3b (15 mg, 38%). Rf (PE / EtOAc = 9:1) = 0.49.1 1H NMR (700 MHz, CDCl3): δ 8.04 (dd, J = 8.0, 1.0 Hz, 2H), 7.56 (tt, J = 7.4, 1.2 Hz, 1H), 7.43 (td, J = 8.0 Hz, 2H), 4.53 (b, 1H), 4.31 (t, J = 6.7 Hz, 2H), 3.10 (b, 2H), 1.76 (quintet, J = 7.2 Hz, 2H), 1.44 (b, 11H), 1.35 (quintet, J = 7.2 Hz, 2H), 1.27 (b, 12H); 13 13C NMR (176 MHz, CD3OD): δ 166.6, 155.9, 132.7, 130.5, 129.5, 128.3, 65.1, 40.6, 30.0, 29.7, 29.5, 29.4, 29.2, 28.7, 28.4, 26.8, 26.0; HRMS (ESI-TOF) m / z: [M + H - Boc] + C 18 H 30 Calculated value for C18H26N2O2: 292.2277, found 292.2269.

[0145] Synthesis of 3b. Benzyl bromide (25 μL, 0.21 mmol) was added to a mixture of sodium hydride (5 mg, 0.21 mmol) and intermediate 2 (50 mg, 0.175 mmol) which had been stirred in dry N,N-dimethylformamide (1.5 mL) in an ice bath for 15 minutes. The mixture was stirred at room temperature overnight. Then, the mixture was extracted using EtOAc and washed three times with deionized water. The collected organic layer was dried over sodium sulfate. Thereafter, the organic layer was concentrated via a rotary evaporator and then the mixture was purified by silica gel column chromatography (eluting with a gradient of petroleum ether and ethyl acetate) to yield the desired compound 3b (29 mg, 50%). Rf (PE / EtOAc = 9:1) = 0.71 f( PE / EtOAc = 9:1) = 0.71 11H NMR (700 MHz, CD3OD) (characteristic protons): δ 7.35 - 7.32 (m, 4H), 7.28 (m, 1H), 5.30 (s, 1H), 4.50 (s, 2H), 3.46 (t, J = 6.7 Hz, 2H); 3.10 (q, J = 6.1 Hz, 2H); 1.61 (quintet, J = 7.1 Hz, 2H); 1.44 (s, 11H), 1.35 (quintet, J = 7.0 Hz, 2H), 1.31 - 1.22 (m, 12H); 13 13C NMR (176 MHz, CD3OD): δ 155.9, 138.7, 128.3, 127.6, 127.4, 72.8, 70.5, 53.4, 40.6, 30.0, 29.73, 29.67, 29.52, 29.50, 29.47, 29.44, 29.3, 28.4, 26.8, 26.2; HRMS (ESI-TOF) m / z: [M + H - Boc] + C 18 H 32 Calculated value for C18H28NO is 278.2484, found 278.2480.

[0146] Synthesis of 4a. Trifluoroacetic acid (85 μl, 30%, v / v) was added to a mixture of 3a (20 mg, 0.07 mmol) and dry dichloromethane (1 mL) to deprotect the Boc group. The mixture was stirred for 3 hours and neutralized with Na2CO3. The mixture was concentrated on a rotary evaporator to give the product (6.5 mg, 33%).

[0147] Synthesis of 4b. Trifluoroacetic acid (25 μl, 30% (v / v)) was added to a mixture of 3b (6 mg, 0.016 mmol) and dry dichloromethane (0.5 mL). The reaction mixture was concentrated and washed with Na2CO3. It was concentrated on a rotary evaporator to give the product 4b (12 mg, 82%).

[0148] General procedure for derivatization of MS II (not optimized): Ethanol / water was added to MS II, and the side chain, N-methylmorpholine (NMM), hydroxybenzotriazole (HOBt), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl) were added at room temperature. The reaction mixture was stirred for 3 days and then semi-Prep C 18 , 250 x 10 mm, 5 micron column, and RP HPLC was used for direct purification by using an H2O / MeCN gradient (90% - 10% H2O over 45 minutes at a flow rate of 3 mL / min). The product fraction was concentrated at room temperature on a rotary evaporator to remove MeCN, and then the remaining water was removed by a freeze dryer to provide the derivative as a white solid.

[0149] For 1D, (12.4 mg, 35%): 1 H NMR (500 MHz, CD3OD) (characteristic protons): δ 9.50 (s, 1H), 8.03 (dd, J = 8.4, 1.3 Hz, 2H), 7.87 (t, J = 5.7 Hz, 1H), 7.63 (tt, J = 7.4, 1.3 Hz, 1H), 7.50 (t, J = 7.8 Hz, 2H), 5.43 (d, J = 1.5 Hz, 1H), 5.33 (b, 1H), 5.23 (d, J = 8.2 Hz, 1H), 5.04 (d, J = 1.4 Hz, 1H), 4.73 (d, J = 7.9 Hz, 1H), 4.56 (d, J = 7.8 Hz, 1H), 4.52 (b, 1H), 4.50 - 4.40 (m, 3H), 4.35 (t, J = 6.6 Hz, 2H), 4.24 (dd, J = 2.8, 1.9 Hz, 1H), 4.06 - 4.01 (m, 2H), 2.30 (t, J = 13.6 Hz, 1H), 1.42 (s, 3H), 1.19 (s, 3H), 1.03 (s, 3H), 0.94 (s, 3H), 0.88 (s, 3H), 0.80 (s, 3H); 1313C NMR (176.0 MHz, CD3OD): δ 209.6, 175.5, 169.8, 166.7, 143.6, 132.8, 130.2, 129.1, 128.2, 121.5, 104.6, 103.7, 103.5, 102.8, 101.9, 99.3, 94.0, 87.4, 84.6, 84.5, 82.2, 77.3, 76.8, 76.5, 76.0, 75.4, 75.1, 74.2, 74.0, 73.6, 73.3, 73.0, 72.4, 72.3, 71.5, 70.8, 70.6, 70.5, 70.0, 68.0, 67.4, 65.7, 64.9, 60.8, 60.6, 54.8, 41.5, 41.0, 39.7, 38.7, 38.0, 35.7, 35.2, 32.0, 30.5, 29.9, 29.5, 29.3, 29.2, 29.0, 28.9, 28.4, 26.6, 26.0, 25.8, 24.6, 23.3, 23.0, 20.0, 17.0, 16.5, 16.4, 15.1, 9.6; HRMS (ESI-TOF) m / z: [M+H] + C 94 H 148 NO 42 Calculated value for 1962.9476, measured value 1962.9467.

[0150] At 1E (8.6 mg, 56% from 1H including benzoyl N-hydroxysuccinimide ester): 11H NMR (600 MHz, CD3OD) (Characteristic protons): δ 9.49 (s, 1H), 7.83 (d, J = 7.4 Hz, 2H), 7.54 (tt, J = 7.4, 1.2 Hz, 1H), 7.48 (d, J = 7.4 Hz, 2H), 5.43 (d, J = 1.6 Hz, 1H), 5.33 (t, J = 3.7 Hz, 1H), 5.23 (d, J = 8.1 Hz, 1H), 5.04 (d, J = 1.7 Hz, 1H), 4.74 (d, J = 7.9 Hz, 1H), 4.56 (d, J = 7.7 Hz, 1H), 4.53 (b, 1H), 4.50 - 4.46 (m, 2H), 4.45 (d, J = 7.7 Hz, 1H), 4.25 (dd, J = 2.9, 2.2 Hz, 1H), 4.06 - 4.01 (m, 2H), 3.18 (t, J = 11.3 Hz, 1H), 3.14 (dd, J = 9.4, 8.1 Hz, 1H), 2.90 (dd, J = 14.6, 4.5 Hz, 1H), 2.30 (t, J = 13.7 Hz, 1H), 1.18 (s, 3H), 1.03 (s, 3H), 0.93 (s, 3H), 0.88 (s, 3H), 0.81 (s, 3H); 13 13C NMR (150.9 MHz, CD3OD): δ 209.6, 175.6, 169.8, 168.8, 143.5, 134.5, 131.1, 128.2, 126.8, 121.6, 104.7, 103.8, 103.5, 102.8, 102.7, 101.9, 99.3, 94.0, 87.4, 84.6, 84.5, 82.2, 77.3, 76.8, 76.4, 76.0, 75.4, 75.1, 74.2, 74.0, 73.6, 73.4, 73.0, 72.4, 72.3, 71.6, 71.5, 70.8, 70.7, 70.5, 70.1, 70.0, 69.6, 69.2, 69.1, 68.0, 67.4, 65.7, 60.8, 54.8, 48.5, 46.9, 46.6, 41.5, 41.0, 39.7, 38.7, 38.0, 35.7, 35.2, 32.8, 31.9, 30.5, 29.9, 29.4, 29.3, 29.1, 28.9, 26.7, 26.5, 25.9, 24.5, 23.3, 23.1, 20.0, 17.0, 16.5, 16.4, 15.1, 9.6; HRMS (ESI-TOF) m / z: [M + H] + C 93 H 147 N2O 41Calculated value for it is 1947.9479, measured value is 1947.9386.

[0151] At 1F (8.3 mg, 36%): 1 HNMR (600 MHz, CD3OD) (characteristic protons): δ 9.50 (s, 1H), 7.37 - 7.34 (m, 4H), 7.30 (m, 1H), 5.43 (d, J = 1.5 Hz, 1H), 5.33 (t, J = 3.3 Hz, 1H), 5.30 (d, J = 8.3 Hz, 1H), 5.04 (d, J = 1.6 Hz, 1H), 4.74 (d, J = 8.1 Hz, 1H), 4.56 (d, J = 8.0 Hz, 1H), 4.25 (dd, J = 2.9, 1.7 Hz, 1H), 2.91 (dd, J = 14.2, 4.5 Hz, 1H), 2.30 (t, J = 13.6 Hz, 1H), 1.19 (s, 3H), 1.03 (s, 3H), 0.94 (s, 3H), 0.88 (s, 3H), 0.81 (s, 3H); 13 C NMR (176.0 MHz, CD3OD): δ 209.6, 175.5, 169.7, 143.6, 138.5, 121.5, 104.7, 103.8, 103.5, 102.8, 102.7, 101.9, 99.3, 94.0, 87.4, 84.6, 84.5, 82.2, 77.2, 76.8, 76.4, 76.0, 75.4, 75.1, 74.2, 74.0, 73.6, 73.3, 73.0, 72.5, 72.4, 72.3, 71.5, 71.4, 70.8, 70.7, 70.5, 70.0, 69.6, 69.2, 69.1, 68.0, 67.4, 65.7, 60.8, 60.6, 54.8, 48.4, 41.5, 41.0, 39.7, 38.7, 35.7, 35.2, 32.7, 32.0, 29.9, 29.5, 29.4, 29.3, 29.2, 28.9, 26.6, 25.9, 23.3, 17.0, 16.5, 16.4, 15.1, 9.6; HRMS (ESI TOF) m / z: [M + H] + C 94 H 150 N O 41 Calculated value for it is 1948.9383, measured value is 1948.9684.

[0152] At 1G (7.4 mg from 1F, 67%): 11H NMR (600 MHz, CD3OD) (characteristic protons): δ 9.49 (s, 1H), 5.42 (d, J = 1.6 Hz, 1H), 5.32 (t, J = 3.5 Hz, 1H), 5.22 (d, J = 8.3 Hz, 1H), 5.03 (d, J = 1.4 Hz, 1H), 4.73 (d, J = 7.8 Hz, 1H), 4.55 (d, J = 7.8 Hz, 1H), 4.52 (b, 1H), 4.44 (d, J = 7.6 Hz, 1H), 4.23 (dd, J = 2.8, 1.8 Hz, 1H), 3.17 (t, J = 10.9 Hz, 1H), 3.12 (dd, J = 8.2, 8.1 Hz, 1H), 2.90 (dd, J = 14.0, 4.4 Hz, 1H), 2.30 (t, J = 13.6 Hz, 1H), 1.42 (s, 3H), 1.19 (s, 3H), 1.03 (s, 3H), 0.95 (s, 3H), 0.88 (s, 3H), 0.80 (s, 3H); 13 13C NMR (213.8 MHz, CD3OD): δ 209.6, 175.6, 169.8, 143.5, 121.5, 104.7, 103.8, 103.6, 102.8, 102.7, 101.9, 99.3, 94.0, 87.4, 84.6, 84.5, 82.1, 77.2, 76.9, 76.4, 76.0, 75.4, 75.1, 74.2, 74.0, 73.6, 73.3, 73.0, 72.4, 72.3, 71.7, 71.5, 70.8, 70.7, 70.5, 70.1, 70.0, 69.6, 69.2, 69.1, 68.0, 67.4, 65.7, 61.6, 60.8, 60.6, 54.8, 46.6, 41.5, 41.0, 39.7, 38.7, 38.0, 35.7, 35.2, 32.7, 32.3, 32.0, 31.7, 30.5, 29.9, 29.5, 29.4, 29.3, 29.2, 29.1, 28.9, 26.5, 25.9, 25.6, 24.5, 23.3, 23.1, 22.3, 20.0, 17.0, 16.5, 16.4, 15.1, 9.6; HRMS (ESI-TOF) m / z: [M+H]+ C 87 H 144 NO 41 Calculated value for 1858.9214, measured value 1858.9211.

[0153] For 1H (20.4 mg, 27%): 11H NMR (600 MHz, CD3OD) (characteristic protons): δ 9.50 (s, 1H), 5.42 (d, J = 1.1 Hz, 1H), 5.33 (t, J = 3.3 Hz, 1H), 5.24 (d, J = 8.4 Hz, 1H), 5.04 (d, J = 1.4 Hz, 1H), 4.74 (d, J = 8.0 Hz, 1H), 4.56 (d, J = 8.0 Hz, 1H), 4.53 (b, 1H), 4.50 - 4.46 (m, 2H), 4.45 (d, J = 7.7 Hz, 1H), 4.25 (dd, J = 3.1, 1.9 Hz, 1H), 4.05 - 4.01 (m, 2H), 3.18 (t, J = 11.1 Hz, 1H), 3.14 (dd, J = 9.4, 8.1 Hz, 1H), 2.31 (t, J = 13.6 Hz, 1H), 1.20 (s, 3H), 1.03 (s, 3H), 0.96 (s, 3H), 0.89 (s, 3H), 0.81 (s, 3H); 13 13C NMR (150.9 MHz, CD3OD): δ 209.6, 175.6, 169.8, 143.6, 121.5, 104.7, 103.8, 103.5, 102.8, 101.7, 101.9, 99.3, 94.0, 87.4, 84.6, 82.1, 77.3, 76.9, 76.4, 76.0, 75.4, 75.1, 74.3, 74.0, 73.6, 73.3, 73.0, 72.4, 72.3, 71.7, 71.5, 70.8, 70.6, 70.5, 70.1, 70.0, 69.6, 69.2, 69.1, 68.0, 67.4, 65.7, 60.8, 60.7, 54.8, 48.5, 46.6, 41.5, 41.0, 39.7, 39.4, 38.7, 37.9, 35.7, 35.2, 32.7, 32.0, 30.5, 29.9, 29.2, 29.0, 28.9, 28.8, 27.2, 26.4, 26.0, 25.9, 24.5, 23.2, 23.1, 20.0, 17.0, 16.5, 16.4, 15.1, 15.0, 9.6; HRMS (ESI-TOF) m / z: [M+H] + C 86 H 143 N2O 40 Calculated value for 1843.9217, measured value 1843.9216.

[0154] Example 2

[0155] [Chemistry]

[0156] General procedure for preparing side chains 5D - 5H (not optimized): A mixture of 11 - aminoundecanoic acid (50 mg, 0.25 mmol) and thionyl chloride (36 μL, 0.5 mmol) was first stirred at room temperature, then heated to 70 °C in an oil bath and stirred for about 1 hour. Substituted benzyl alcohol (0.5 mmol) was added to the reaction mixture and stirred overnight. The mixture was purified by silica gel column chromatography (eluted with a DCM / MeOH gradient) to produce the desired side chain.

[0157] Synthesis of 5D. Using 3,5 - dimethoxybenzyl alcohol (84.0 mg, 0.5 mmol) and purified by silica gel column chromatography (eluted with a DCM / MeOH gradient) to produce 5D (41.0 mg, 47%). R f (DCM / MeOH 9:1) 0.43; 1 1H NMR (700 MHz, CD3OD): δ 6.48 (d, J = 2.2 Hz, 2H), 6.40 (t, J = 2.2 Hz, 1H), 5.03 (s, 2H), 3.77 (s, 6H), 2.97 (t, J = 7.9 Hz, 2H), 2.34 (t, J = 7.6 Hz, 2H), 1.75 (quintet, J = 7.8 Hz, 2H), 1.63 (quintet, J = 7.8 Hz, 2H), 1.35 (quintet, J = 7.3 Hz, 2H), 1.32 - 1.22 (m, 10H); 13 13C NMR (176.0 MHz, CD3OD): δ 173.5, 160.9, 138.4, 105.9, 100.1, 65.9, 55.3, 40.0, 34.3, 29.4, 29.3, 29.2, 29.1, 29.0, 27.6, 26.6, 24.9; HRMS (ESI - TOF) m / z: [M + H] + C 20 H 34 Calculated for C19H31NO4 352.2488, found 352.2486.

[0158] Synthesis of 5E. 3-Methoxybenzyl alcohol (63.0 mg, 0.5 mmol) and purification by silica gel column chromatography (eluting with a DCM / MeOH gradient) yielded 5E (65.0 mg, 81%). R f (DCM / MeOH 9:1) 0.56; 1H NMR (700 MHz, CD3OD): δ 7.26 (d, J = 7.9 Hz, 1H), 6.92 (d, J = 7.3 Hz, 1H), 6.88 (s, 1H), 6.85 (dd, J = 7.7, 2.1 Hz, 1H), 5.08 (s, 2H), 3.80 (s, 3H), 2.97 (t, J = 7.7 Hz, 2H), 2.34 (t, J = 7.9 Hz, 2H), 1.76 (quintet, J = 7.7 Hz, 2H), 1.63 (quintet, J = 7.3 Hz, 2H), 1.37 (quintet, J = 7.3 Hz, 2H), 1.33 - 1.23 (m, 10H); 13 13C NMR (176.0 MHz, CD3OD): δ 173.6, 159.8, 137.7, 129.6, 120.3, 113.6, 65.9, 55.2, 40.0, 34.3, 29.6, 29.4, 29.3, 29.1, 28.9, 27.7, 26.5, 24.9; HRMS (ESI-TOF) m / z: [M + H] + C 19 H 32 Calculated for C19H30NO3 322.2382, found 322.2380.

[0159] Synthesis of 5F: 3-Phenoxyphenylmethanol (87.0 μL, 0.5 mmol) and purification by silica gel column chromatography (eluting with a DCM / MeOH gradient) yielded 5F (50.0 mg, 52%). R f (DCM / MeOH 9:1) 0.43; 11H NMR (700 MHz, CD3OD) δ 7.32 (t, J = 17.0 Hz, 2H), 7.29 (t, J = 7.9 Hz, 1H), 7.10 (t, J = 7.4 Hz, 1H), 7.06 (d, J = 7.7 Hz, 1H), 7.00 - 6.98 (m, 3H), 6.93 (dd, J = 8.2, 2.2 Hz, 1H), 5.07 (s, 3H), 2.97 (t, J = 7.9 Hz, 2H), 2.33 (t, J = 7.5 Hz, 2H), 1.76 (quintet, J = 7.7 Hz, 2H), 1.61 (quintet, J = 7.4 Hz, 2H), 1.37 (quintet, J = 7.4 Hz, 2H), 1.33 - 1.23 (m, 10H); 13 13C NMR (176.0 MHz, CD3OD): δ 173.4, 157.5, 157.0, 138.2, 129.8, 129.6, 123.4, 122.6, 119.0, 118.3, 118.2, 65.5, 40.0, 34.3, 29.7, 29.3, 29.2, 29.1, 29.0, 27.7, 26.6, 24.9; HRMS (ESI-TOF) m / z: [M + H] + C 24 H 34 Calculated for C21H28NO3: 384.2539, Found: 384.2535.

[0160] Synthesis of 5G: 5G (64.0 mg, 76%) was produced by purification of 3-nitrobenzyl alcohol (60.0 mg, 0.5 mmol) by silica gel column chromatography (eluting with a DCM / MeOH gradient). R f (DCM / MeOH 9:1) 0.40; 1 1H NMR (700 MHz, CD3OD): δ 8.21 (s, 1H), 8.13 (d, J = 7.9 Hz, 1H), 7.67 (d, J = 7.6 Hz, 1H), 7.54 (t, J = 7.9 Hz, 1H), 5.20 (s, 2H), 2.96 (t, J = 7.7 Hz, 2H), 2.38 (t, J = 7.6 Hz, 2H), 1.74 (quintet, J = 7.6 Hz, 2H), 1.64 (quintet, J = 7.4 Hz, 2H), 1.36 (quintet, J = 7.3 Hz, 2H), 1.33 - 1.22 (m, 10H); 1313C NMR (176.0 MHz, CD3OD): δ 173.3, 148.4, 138.3, 133.8, 129.5, 123.0, 122.7, 64.5, 40.0, 39.5, 36.8, 34.1, 29.6, 29.4, 29.3, 29.1, 29.0, 27.7, 26.9, 26.5, 26.4, 25.7, 24.8; HRMS (ESI-TOF) m / z: [M+H] + C 18 H 29 Calculated value for N2O4: 337.2127, measured value: 337.2126.

[0161] Synthesis of 5H: 4-Fluorobenzyl alcohol (55.0 μL, 0.5 mmol) and purification by silica gel column chromatography (eluting with a DCM / MeOH gradient) yielded 5H (49.0 mg, 64%). R f (DCM / MeOH 9:1) 0.37; 1 1H NMR (700 MHz, CD3OD): δ 7.31 (dd, J = 8.5, 5.6 Hz, 1H), 7.02 (t, J = 8.5 Hz, 1H), 5.06 (s, 2H), 2.96 (t, J = 7.8 Hz, 2H), 2.31 (t, J = 7.4 Hz, 2H), 1.73 (quintet, J = 7.5 Hz, 2H), 1.60 (quintet, J = 7.2 Hz, 2H), 1.34 (quintet, J = 7.0 Hz, 2H), 1.31 - 1.21 (m, 10H); 13 13C NMR (176.0 MHz, CD3OD): δ 173.5, 163.3, 161.9, 132.0, 130.1, 115.5, 115.3, 65.2, 40.1, 39.5, 36.8, 34.2, 29.6, 29.4, 29.3, 29.2, 29.1, 28.8, 27.7, 26.9, 26.6, 26.4, 25.7, 24.9; HRMS (ESI-TOF) m / z: [M+H] + C 18 H 29 Calculated value for FNO2: 310.2182, measured value: 310.2179.

[0162] General procedure for derivatization of MS II (not optimized): To MS II (15 mg, 0.009 mmol), ethanol / water (1:2, 0.5 mL) was added, and the side chain (5D - 5H) (0.018 mmol), N-methylmorpholine (NMM) (10.0 μl, 0.09 mmol), hydroxylbenzotriazole (HOBt) (8.3 mg, 0.054 mmol), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC HCl) (11.0 mg, 0.054 mmol) were added at room temperature. The reaction mixture was stirred for 3 days and then directly purified by RP HPLC using a semi-Prep C18, 250x10 mm, 5 micron column, and an H2O / MeCN gradient (90% - 10% H2O over 45 minutes at a flow rate of 3 mL / min). The product fraction was concentrated at room temperature on a rotary evaporator to remove MeCN, and then the remaining water was removed by freeze dryer to afford the derivative as a white solid.

[0163] In 2D, 5D (6.3 mg, 0.018 mmol) was used to afford the final product 2D (10 mg, 56%). 1 H NMR (600 MHz, CD3OD) (characteristic protons): δ 9.47 (s, 1H), 6.50 (d, J = 2.3 Hz, 2H), 6.42 (t, J = 2.3 Hz, 1H), 5.41 (d, J = 1.5 Hz, 1H), 5.30 (t, J = 3.6 Hz, 1H), 5.20 (d, J = 8.8 Hz, 1H), 5.05 (s, 2H), 5.02 (d, J = 1.4 Hz, 1H), 4.72 (d, J = 7.8 Hz, 1H), 4.56 (d, J = 8.0 Hz, 1H), 4.53 (d, J = 8.1 Hz, 1H), 4.50 (b, 1H), 4.44 (d, J = 7.6 Hz, 1H), 4.23 (dd, J = 3.0, 1.8 Hz, 1H), 3.15 (t, J = 11.0 Hz, 1H), 3.13 (dd, J = 9.4, 8.1 Hz, 1H), 2.88 (dd, J = 14.1, 4.5 Hz, 1H), 2.38 (t, J = 7.3 Hz, 2H), 2.27 (t, J = 13.4 Hz, 1H), 1.16 (s, 3H), 0.99 (s, 3H), 0.92 (s, 3H), 0.86 (s, 3H), 0.77 (s, 3H); 1313C NMR (176.0 MHz, CD3OD): δ 209.6, 175.5, 173.8, 169.8, 161.0, 143.5, 138.6, 121.5, 105.5, 104.7, 103.8, 103.5, 102.8, 101.9, 99.5, 99.3, 94.0, 87.4, 84.6, 84.5, 82.2, 77.3, 76.8, 76.4, 76.0, 75.4, 75.1, 74.2, 74.0, 73.6, 73.4, 73.0, 72.4, 72.3, 71.5, 70.8, 70.7, 70.5, 70.1, 70.0, 69.6, 69.2, 69.1, 68.0, 67.4, 65.7, 65.6, 60.8, 60.6, 54.8, 54.4, 41.5, 41.0, 39.7, 38.7, 38.0, 35.7, 35.3, 33.7, 32.8, 32.0, 30.5, 29.9, 29.4, 29.2, 29.1, 29.0, 28.7, 26.5, 25.9, 24.7, 23.3, 23.1, 20.0, 17.0, 16.5, 16.4, 15.1, 9.6; HRMS (ESI-TOF) m / z: [M+H] + C 96 H 152 NO 44 Calculated value for 2022.9687, measured value 2022.9675.

[0164] In 2E, 5E (6 mg, 0.018 mmol) was used to afford the final product 2E (8 mg, 45%). 11H NMR (600 MHz, CD3OD): δ 9.47 (s, 1H), 7.26 (t, J = 7.9 Hz, 1H), 6.93 - 6.89 (m, 2H), 6.87 (dd, J = 8.2, 2.7 Hz, 1H), 5.40 (d, J = 1.8 Hz, 1H), 5.30 (t, J = 3.7 Hz, 1H), 5.20 (d, J = 8.2 Hz, 1H), 5.09 (s, 2H), 5.01 (d, J = 1.6 Hz, 1H), 4.71 (d, J = 7.9 Hz, 1H), 4.53 (d, J = 7.9 Hz, 1H), 4.49 (b, 1H), 4.42 (d, J = 7.7 Hz, 1H), 4.22 (dd, J = 3.1, 1.9 Hz, 1H), 3.15 (t, J = 10.8 Hz, 1H), 3.11 (dd, J = 9.3, 8.0 Hz, 1H), 2.88 (dd, J = 14.3, 4.1 Hz, 1H), 2.37 (t, J = 7.3 Hz, 2H), 2.27 (t, J = 13.6 Hz, 1H), 1.39 (s, 3H), 1.22 (d, J = 6.3 Hz, 3H), 1.20 (d, J = 6.5 Hz, 3H), 1.16 (s, 3H), 1.00 (s, 3H), 0.91 (s, 3H), 0.85 (s, 3H), 0.78 (s, 3H); 13 13C NMR (176.0 MHz, CD3OD): δ 209.6, 175.5, 173.8, 169.8, 161.4, 160.0, 143.5, 137.8, 129.2, 121.5, 119.9, 113.2, 104.7, 103.8, 103.5, 102.8, 102.7, 101.9, 99.3, 94.0, 87.4, 84.6, 84.5, 82.2, 77.3, 76.8, 76.4, 76.0, 75.5, 75.4, 75.1, 74.2, 74.0, 73.6, 73.3, 73.0, 72.4, 72.3, 71.5, 79.8, 70.7, 70.5, 70.1, 70.0, 69.6, 69.2, 69.1, 68.0, 65.7, 65.6, 60.8, 54.8, 54.3, 41.5, 39.7, 38.8, 35.7, 35.3, 33.7, 32.0, 29.9, 29.4, 29.2, 29.1, 29.0, 28.9, 28.7, 26.5, 25.9, 24.7, 23.3, 17.0, 16.5, 16.4, 15.1, 9.6; HRMS(ESI-TOF) m / z: [M + H]+ C 95 H 149 NO 43Calculated value for it is 1992.9582, measured value is 1992.9662.

[0165] In 2F, 5F (7 mg, 0.018 mmol) was used to produce the final product 2F (10.0 mg, 54%): 1 H NMR (600 MHz, CD3OD): δ 9.51 (s, 1H), 7.42 - 7.30 (m, 3H), 7.17 (tt, J = 7.4, 2.1 Hz, 1H), 7.14 (d, J = 3.7 Hz, 1H), 7.05 - 7.01 (m, 3H), 6.97 (dd, J = 8.1, 2.3 Hz, 1H), 5.45 (d, J = 1.6 Hz, 1H), 5.34 (t, J = 3.5 Hz, 1H), 5.25 (d, J = 8.3 Hz, 1H), 5.14 (s, 2H), 5.06 (d, J = 1.5 Hz, 1H), 4.76 (d, J = 7.9 Hz, 1H), 4.58 (d, J = 7.9 Hz, 1H), 4.55 (b, 1H), 4.47 (d, J = 7.7 Hz, 1H), 4.27 (dd, J = 3.0, 1.9 Hz, 1H), 4.07 - 4.04 (m, 2H), 3.20 (t, J = 11.0 Hz, 1H), 3.16 (dd, J = 9.4, 8.0 Hz, 1H), 2.93 (dd, J = 14.1, 4.4 Hz, 1H), 2.40 (t, J = 7.3 Hz, 2H), 2.32 (t, J = 13.6 Hz, 1H), 1.44 (s, 3H), 1.27 (d, J = 6.2 Hz, 3H), 1.25 (d, J = 6.4 Hz, 3H), 1.21 (s, 3H), 1.04 (s, 3H), 0.96 (s, 3H), 0.90 (s, 3H), 0.82 (s, 3H); 1313C NMR (150.9 MHz, CD3OD): δ 209.6, 175.5, 173.7, 169.8, 157.6, 157.1, 138.6, 129.7, 129.6, 123.2, 122.4, 121.5, 118.7, 117.9, 117.7, 104.7, 103.8, 103.5, 102.8, 101.9, 99.3, 94.0, 87.4, 84.6, 84.5, 82.2, 77.3, 76.8, 76.4, 76.0, 75.4, 75.1, 74.3, 74.2, 74.0, 73.6, 73.4, 73.0, 72.4, 72.3, 71.5, 70.8, 70.7, 70.5, 70.1, 70.0, 69.6, 69.2, 69.1, 68.0, 67.4, 65.7, 65.2, 60.8, 60.6, 54.8, 41.5, 41.0, 39.7, 38.7, 38.0, 35.7, 35.3, 33.7, 32.0, 30.5, 29.9, 29.4, 29.2, 29.1, 29.0, 28.9, 28.7, 26.5, 26.0, 24.7, 23.3, 23.1, 20.0, 17.0, 16.5, 16.4, 15.1, 9.6; HRMS (ESI-TOF) m / z: [M+H] + C 100 H 152 NO 43 Calculated value for 2054.9738, measured value 2054.9714.

[0166] In 2G, using 5G (6 mg, 0.018 mmol), the final product 2G (10.0 mg, 56%) was produced: 11H NMR (600 MHz, CD3OD): δ 9.51 (s, 1H), 8.29 (s, 1H), 8.24 (dd, J = 8.2, 1.9 Hz, 1H), 7.82 (d, J = 7.5 Hz, 1H), 7.68 (t, J = 7.9 Hz, 1H), 5.44 (d, J = 1.5 Hz, 1H), 5.33 (b, 1H), 5.29 (s, 2H), 5.23 (d, J = 8.2 Hz, 1H), 5.05 (d, J = 1.2 Hz, 1H), 4.75 (d, J = 7.8 Hz, 1H), 4.57 (d, J = 7.8 Hz, 1H), 4.54 (b, 1H), 4.46 (d, J = 7.7 Hz, 1H), 4.26 (b, 1H), 4.07 - 4.02 (m, 2H), 3.19 (t, J = 11.7 Hz, 1H), 3.15 (dd, J = 9.4, 8.0 Hz, 1H), 2.91 (dd, J = 14.0, 4.4 Hz, 1H), 2.45 (t, J = 7.3 Hz, 2H), 2.30 (t, J = 13.9 Hz, 1H), 1.43 (s, 3H), 1.26 (d, J = 6.3 Hz, 3H), 1.24 (d, J = 6.5 Hz, 3H), 1.20 (s, 3H), 1.03 (s, 3H), 0.94 (s, 3H), 0.89 (s, 3H), 0.81 (s, 3H); 13 13C NMR (176.0 MHz, CD3OD): δ 209.6, 175.5, 173.5, 169.6, 148.3, 143.6, 138.8, 133.9, 129.6, 124.4, 122.6, 122.3, 121.5, 104.7, 103.7, 103.5, 102.8, 101.9, 99.3, 94.0, 87.4, 84.6, 84.5, 82.1, 77.3, 76.8, 76.4, 76.0, 75.4, 75.1, 74.2, 74.0, 73.6, 73.3, 73.0, 72.4, 72.3, 71.5, 70.8, 70.7, 70.5, 70.1, 70.0, 69.6, 69.2, 69.1, 68.0, 67.4, 65.7, 64.4, 60.8, 60.6, 54.8, 41.5, 41.0, 39.7, 38.7, 38.0, 35.7, 35.3, 33.6, 32.8, 32.0, 30.5, 29.9, 29.4, 29.2, 29.1, 28.9, 28.8, 26.5, 25.9, 24.7, 23.3, 23.1, 20.0, 17.0, 16.5, 16.4, 15.1, 9.6; HRMS (ESI-TOF) m / z: [M + H]+ C 94 H 147 N2O 44 Calculated value for N2O: 2007.9327, measured value: 2007.9337.

[0167] For 2H, 5H (5.6 mg, 0.018 mmol) was used to produce the final product 2H (10.0 mg, 56%): 1 H NMR (600 MHz, CD3OD): δ 9.51 (s, 1H), 7.45 - 7.40 (m, 2H), 7.11 (tt, J = 8.7, 2.1 Hz, 1H), 5.44 (d, J = 1.6 Hz, 1H), 5.34 (t, J = 3.4 Hz, 1H), 5.24 (d, J = 8.3 Hz, 1H), 5.13 (s, 2H), 5.05 (d, J = 1.6 Hz, 1H), 4.75 (d, J = 8.0 Hz, 1H), 4.57 (d, J = 7.8 Hz, 1H), 4.54 (b, 1H), 4.46 (d, J = 7.6 Hz, 1H), 4.26 (dd, J = 3.0, 2.1 Hz, 1H), 3.19 (t, J = 11.0 Hz, 1H), 3.15 (dd, J = 9.2, 8.3 Hz, 1H), 2.92 (dd, J = 14.5, 4.4 Hz, 1H), 2.39 (t, J = 7.5 Hz, 2H), 2.31 (t, J = 13.6 Hz, 1H), 1.44 (s, 3H), 1.26 (d, J = 6.3 Hz, 3H), 1.25 (d, J = 6.4 Hz, 3H), 1.21 (s, 3H), 1.04 (s, 3H), 0.95 (s, 3H), 0.89 (s, 3H), 0.82 (s, 3H); 1313C NMR (176.0 MHz, CD3OD): δ 209.6, 175.5, 173.7, 169.8, 163.3, 161.9, 143.6, 132.5, 130.2, 130.1, 114.9, 114.8, 104.7, 103.7, 103.5, 102.8, 101.9, 99.3, 94.0, 87.4, 84.6, 84.5, 82.2, 77.3, 76.8, 76.4, 76.0, 75.4, 75.1, 74.2, 74.0, 73.6, 73.3, 73.0, 72.4, 72.3, 71.5, 70.8, 70.7, 70.5, 70.1, 70.0, 69.6, 69.2, 69.1, 68.0, 67.4, 65.7, 65.0, 60.8, 60.6, 54.8, 48.5, 41.5, 41.0, 39.7, 38.7, 38.0, 35.7, 35.3, 33.7, 32.8, 32.0, 30.5, 29.9, 29.4, 29.2, 29.1, 29.0, 28.9, 28.7, 26.5, 25.9, 24.7, 23.3, 23.1, 20.0, 17.0, 16.5, 16.4, 15.1, 9.6; HRMS (ESI-TOF) m / z: [M+H] + C 94 H 147 FNO 42 Calculated value for 1980.9382, measured value 1980.9463.

[0168] Antigen: Chicken ovalbumin (Vac-pova) for in vivo use was purchased from InvivoGen.

[0169] Mice and immunization: The BALB / c mice used in this study were from Frederick Cancer Research (Frederick, MD). To evaluate the adjuvant activity of the MS saponin-based immunoadjuvant, female mouse groups (8 - 10 weeks old, 6 mice per group) were immunized via the subcutaneous (s.c.) route on days 0, 14, and 28 with OVA (20 μg) alone, or with an appropriate adjuvant such as QS-21 (20 μg) or the MS adjuvant (50 - 100 μg) as antigen. Before each immunization and 2 weeks after the last immunization, the mice were weighed and blood samples were collected from the lateral tail vein by using heparinized capillary pipettes. Serum was obtained after centrifugation and stored at -20 °C until assay.

[0170] Evaluation of antibody response: The levels of specific serum IgG and IgG subclasses against OVA in each group were determined by enzyme-linked immunosorbent assay (ELISA). Maxisorpmicrotiter plates (NUNC International, Roskilde, DK) were coated overnight at 4°C with rHagB (1 μg / ml), OVA (0.1 μg / ml), or optimal amounts of goat anti-mouse IgG, IgG1, or IgG2a in borate buffered saline (BBS; 100 mM NaCl, 50 mM boric acid, 1.2 mM Na2B4O7, pH 8.2). The plates were blocked for 2 hours at room temperature with 1% bovine serum albumin (BSA) and 0.02% sodium azide in BBS. Serial two-fold dilutions of serum samples were added in duplicate to the plates. To generate a standard curve, serial dilutions of mouse immunoglobulin reference serum (MP Biomedicals, Solon, OH) were added to two columns of wells of each plate coated with the appropriate anti-mouse IgG or IgG subclass reagent. After incubation (overnight at 4°C) and washing of the plates, horseradish peroxidase-conjugated goat anti-mouse IgG or IgG subclass antibody was added to the appropriate wells. After incubation for 4 hours at room temperature, the plates were washed and developed with o-phenylenediamine substrate containing hydrogen peroxide. Color development was recorded at 490 nm. The antibody concentration was determined by interpolation against a standard curve generated using mouse immunoglobulin reference serum and constructed by a computer program based on a four-parameter logistic algorithm (Softmax / Molecular Devices Corp., Menlo Park, CA).

[0171] Figures 5A - 5C illustrate the serum IgG, IgG1, and IgG2a anti-OVA responses in mice immunized via the subcutaneous route with ovalbumin (OVA) alone, or with QS-21 or VSA-2, or one of saponin adjuvants 1D - 1H. Statistical significance was evaluated by t-test (using non-parametric and Mann-Whiteny tests where appropriate). *P < 0.05, **P < 0.01.

[0172] Figures 6A - 6C illustrate the serum IgG, IgG1, and IgG2a anti - OVA responses in mice immunized subcutaneously with ovalbumin (OVA) alone or with one of QS - 21 or VSA - 2, or saponin adjuvants 2D - 2H.

[0173] Statistical analysis: Statistical significance in antibody responses was evaluated by t - test using GraphPad Prism 8 (using non - parametric and Mann - Whiteny tests where applicable). A difference with a P - value < 0.05 was considered significant.

[0174] Conclusion: A number of MS saponin - derived immunostimulants were prepared and evaluated. These MS derivatives were prepared by incorporating terminal functionalized side chains into the C3 glucuronic acid unit of natural saponin MSII through an amide - forming reaction. These non - natural semi - synthetic saponins showed an immunostimulatory activity profile as potent as that of the FDA - approved saponin immunostimulant QS - 21.

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Claims

1. Modified saponins having the following formula: 【Chemistry 1】 (In the formula, q １ However, it is either H or OH, q ２ and q ３ However, each operates independently as CHO, CH ３ ,CH ２ Selected from OH, H, COOH, acetal group components, or imine groups, f ３ and f ４ are each independently OH, or acetyl, or C3 and C4 of a fucosyl unit, and f ３ and f ４ can form a cyclic ketal ring or a cyclic carbonate ester f ５ However, H, methyl group, -CH ２ OH group, R ４ -NR ５ -C(O)-, R ４ -O-C(O)-, or R ４ -O-CH ２ - Selected from, R ４ and R ５ However, each independently, Structure R ６ [(CX ２ ) ０－２０ O ０－１ (CY ２ ) ０－２０ ] ０－２０ It is a linear chain having, X and Y are each independently H or a halogen atom. R ６ However, COOR ７ , C(O)NR ７ R ８ , NR ７ R ８ , or OR ７ And, R ７ However, Ar[(CZ ２ ) ０－２０ O ０－１ (CL ２ ) ０－２０ ] ０－２０ And R ８ However, it is H or alkyl, Z and L are each independently H or a halogen atom. Ar is a substituted or unsubstituted aromatic system. ga ５ However, R １４ -NR １５ -C(O)-, R １４ -O-C(O)-, or R １４ -O-CH ２ - Selected from, R １４ and R １５ However, each independently, Structure R １６ [(CX1 ２ ) ０－２０ O ０－１ (CY1 ２ ) ０－２０ ] ０－２０ It is a linear chain having, X1 and Y1 are each independently H or a halogen atom. R １６ However, COOR １７ , C(O)NR １７ R １８ , NR １７ R １８ , or OR １７ And, R １７ However, Ar2[(CZ1 ２ ) ０－２０ O ０－１ (CL1 ２ ) ０－２０ ] ０－２０ And R １８ However, it is H or alkyl, Z1 and L1 are each independently H or a halogen atom. Ar2 is a substituted or unsubstituted aromatic group. r3 is H, monosaccharide, dissaccharide, or trisaccharide. x3 is H, monosaccharide (excluding xylose), or dissaccharide, and (ga3 is H, a monosaccharide, or a dissaccharide.)

2. Modified saponins having the following formula I: 【Chemistry 2】 (In the formula, q １ However, it is either H or OH, q ２ and q ３ However, each operates independently as CHO, CH ３ ,CH ２ Selected from OH, H, COOH, acetal group components, or imine groups, ga ５ However, R １４ -NR １５ -C(O)-, R １４ -O-C(O)-, or R １４ -O-CH ２ - Selected from, R １４ and R １５ are each independently a straight chain having a structure R １６ [(CX1 ２ ) ０－２０ O ０－１ (CY1 ２ ) ０－２０ ０－２０ and are straight chains​ X1 and Y1 are each independently H or a halogen atom. R １６ is COOR １７ C(O)NR １７ R １８ NR １７ R １８ or OR １７ and R １７ However, Ar2[(CZ1 ２ ) ０－２０ O ０－１ (CL1 ２ ) ０－２０ ] ０－２０ And R １８ However, it is H or alkyl, Z1 and L1 are each independently H or a halogen atom, and Ar2 is a substituted or unsubstituted aromatic group.

3. A modified saponin having one of the following formulas 2D to 2H. 【Transformation 3】

4. A pharmaceutical composition comprising a therapeutically effective amount of a modified saponin having the following formula for treating a medical condition: 【Chemistry 4】 (In the formula, q １ However, it is either H or OH, q ２ and q ３ However, each operates independently as CHO, CH ３ ,CH ２ Selected from OH, H, COOH, acetal group components, or imine groups, f ３ and f ４ However, each is independently a C3 and C4 unit of OH, or acetyl, or fuocsyl, and f ３ and f ４ However, it can form a cyclic ketal ring or a cyclic carbonate ester. f ５ However, H, methyl group, -CH ２ OH group, R ４ -NR ５ -C(O)-, R ４ -O-C(O)-, or R ４ -O-CH ２ - Selected from, R ４ and R ５ However, each independently, Structure R ６ [(CX ２ ) ０－２０ O ０－１ (CY ２ ) ０－２０ ] ０－２０ It is a linear chain having, X and Y are each independently H or a halogen atom. R ６ However, COOR ７ , C(O)NR ７ R ８ , NR ７ R ８ , or OR ７ And, R ７ However, Ar[(CZ ２ ) ０－２０ O ０－１ (CL ２ ) ０－２０ ] ０－２０ And R ８ However, it is H or alkyl, Z and L are each independently H or a halogen atom. Ar is a substituted or unsubstituted aromatic system. ga ５ However, R １４ -NR １５ -C(O)-, R １４ -O-C(O)-, or R １４ -O-CH ２ - Selected from, R １４ and R １５ However, each independently, Structure R １６ [(CX1 ２ ) ０－２０ O ０－１ (CY1 ２ ) ０－２０ ] ０－２０ It is a linear chain having, X1 and Y1 are each independently H or a halogen atom. R １６ However, COOR １７ , C(O)NR １７ R １８ , NR １７ R １８ , or OR １７ And, R １７ However, Ar2[(CZ1 ２ ) ０－２０ O ０－１ (CL1 ２ ) ０－２０ ] ０－２０ And R １８ However, it is H or alkyl, Z1 and L1 are each independently H or a halogen atom. Ar2 is a substituted or unsubstituted aromatic group. r3 is H, monosaccharide, dissaccharide, or trisaccharide. x3 is H, monosaccharide (excluding xylose), or dissaccharide, and (ga3 is H, a monosaccharide, or a dissaccharide.)

5. A pharmaceutical composition comprising a therapeutically effective amount of a modified saponin having the following formula I for treating a medical condition: 【Transformation 5】 (In the formula, q １ However, it is either H or OH, q ２ and q ３ However, each operates independently as CHO, CH ３ ,CH ２ Selected from OH, H, COOH, acetal group components, or imine groups, ga ５ However, R １４ -NR １５ -C(O)-, R １４ -O-C(O)-, or R １４ -O-CH ２ - Selected from, R １４ and R １５ However, each independently, Structure R １６ [(CX1 ２ ) ０－２０ O ０－１ (CY1 ２ ) ０－２０ ] ０－２０ It is a linear chain having, X1 and Y1 are each independently H or a halogen atom. R １６ However, COOR １７ , C(O)NR １７ R １８ , NR １７ R １８ , or OR １７ And, R １７ However, Ar2[(CZ1 ２ ) ０－２０ O ０－１ (CL1 ２ ) ０－２０ ] ０－２０ And R １８ However, it is H or alkyl, Z1 and L1 are each independently H or a halogen atom, and Ar2 is a substituted or unsubstituted aromatic group.

6. A pharmaceutical composition comprising a therapeutically effective amount of a modified saponin having one of formulas 2D to 2H for treating a medical condition. 【Transformation 6】

7. A pharmaceutical composition according to any one of claims 4 to 6, further comprising at least one immunogen.

8. A pharmaceutical composition according to any one of claims 4 to 6, further comprising a pharmaceutically acceptable carrier.

9. A pharmaceutical composition according to any one of claims 4 to 6, formulated for administration to animals or humans.

10. The pharmaceutical composition for cancer treatment according to any one of claims 4 to 6, further comprising a pharmaceutically acceptable carrier, wherein at least one chemotherapeutic agent and the modified saponin are mixed in or covalently bonded to each other in a pharmaceutically acceptable formulation.

11. A vaccine for use in methods for increasing the immunogenicity of an immunogen in animals or humans, The vaccine comprises at least one pharmaceutical composition according to any one of claims 4 to 6. The method includes the step of administering at least the vaccine to the subject, vaccine.

12. The pharmaceutical composition further comprises at least one immunogen, The pharmaceutical composition further comprises a pharmaceutically acceptable carrier. The aforementioned pharmaceutical composition is formulated for administration to animals or humans, or The pharmaceutical composition further comprises a pharmaceutically acceptable carrier, wherein at least one chemotherapeutic agent and the modified saponin are mixed in or covalently bonded to each other within a pharmaceutically acceptable formulation, and is a pharmaceutical composition for the treatment of cancer. The vaccine according to claim 11.

13. A method for synthesizing a unique compound according to any one of claims 1 to 3, comprising a synthetic route that includes coupling a natural saponin with a functionalized side chain molecule to provide a ga5 group of the unique compound according to claim 1, 2, or 3.

14. The method according to claim 13, wherein the natural saponin is obtained from Momordica cochinchinensis Spreng.

15. The method according to claim 13, wherein the natural saponin is coupled to the functionalized side chain molecule via an amide formation reaction or an ester formation reaction.

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