Saponin-based immunostimulatory agent, pharmaceutical composition comprising said immunostimulatory agent, therapeutic use thereof

Abstract

The present disclosure provides compounds including modified saponin compounds, pharmaceutical compositions including modified saponin compounds, methods of using the modified saponin compounds and the pharmaceutical compositions, methods of making modified saponin compounds, and the like. The compounds and pharmaceutical compositions of the present disclosure may be used in combination with one or more other therapeutic agents for the treatment of viral infections and other diseases.

Description

[0001] Claiming priority from a related application

[0002] This application claims priority to the co-pending U.S. provisional application entitled “Saponin-based immunostimulants, pharmaceutical composition comprising said immunostimulants, therapeutic use thereof” (serial number: 63 / 364,897), filed on May 18, 2022, which is incorporated herein by reference in its entirety.

[0003] Statement on U.S. Government Funding

[0004] This invention was made with government support under Contract R01 GM120159 awarded by the National Institutes of Health. The government has certain rights in this invention. Background Art

[0005] Vaccine adjuvants are substances used with vaccines to enhance the host's immune response to specific antigens 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).

[0006] Vaccine adjuvants also adjust the immune system to the desired response for certain pathogens. For example, QS-21 (a mixture of two isomers) is an FDA-approved adjuvant known for its ability to enhance the balance of Th1 / Th2 responses using antigen-specific CTL production, which is very valuable 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 a wide range of clinical application potential, and therefore is in great 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 the soap tree (QS), an evergreen tree native to the warm temperate zone of central Chile. However, even under existing demand, overexploitation of natural resources has already caused ecological and economic consequences. In addition, the abundance of QS-21 in QS bark extract is low, and its separation is time-consuming and 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 chemical instability problems due to two hydrolysis-unstable ester parts that complicate its preparation; its dose-limiting toxicity also prevents it from exerting its full effectiveness.

[0007] Derivatization of Momordica saponins (MS) I and II has been shown to be a potential viable approach to obtain 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 fruit (MC) grown in China and Southeast Asia. Derivatization of MS I / II at the C3 glucuronic acid can produce immunostimulants with adjuvant effects similar to those of QS-21. Summary of the invention

[0008] The present disclosure provides compounds including modified saponin compounds, pharmaceutical compositions including modified saponin compounds, methods of using the modified saponin compounds and the pharmaceutical compositions, methods of preparing modified saponin compounds, and the like.

[0009] The present disclosure provides modified saponins having the formula:

[0010]

[0011] in:

[0012] q1 can be H or OH;

[0013] q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a component of an acetal group or an imine group;

[0014] f3 and f4 are each independently OH or acetyl or C3 and C4 of a fucosyl unit, wherein f3 and f4 may form a cyclic ketal ring or a cyclic carbonate;

[0015] f5 is selected from H, a methyl group, a -CH2OH group, R4-NR5-C(O)-, R4-OC(O)- or R4-O-CH2-;

[0016] wherein R4 and R5 are each independently a compound having the structure R6[(CX2) 0-20 O 0-1 (CY2) 0-20 ] 0-20 A linear chain of

[0017] wherein X and Y are each independently H or a halogen atom;

[0018] wherein R6 is COOR7, C(O)NR7R8, NR7R8 or OR7;

[0019] Where R7 is Ar[(CZ2) 0-20 O 0-1 (CL2) 0-20 ] 0-20 , wherein R8 is H or alkyl;

[0020] wherein Z and L are each independently H or a halogen atom;

[0021] wherein Ar is a substituted or unsubstituted aromatic system;

[0022] ga5 is selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-;

[0023] Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12)0-20 ] 0-20 A linear chain of

[0024] wherein X1 and Y1 are each independently H or a halogen atom;

[0025] Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ;

[0026] Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl;

[0027] Wherein Z1 and L1 are each independently H or a halogen atom;

[0028] wherein Ar2 is a substituted or unsubstituted aromatic group;

[0029] r3 is H, a monosaccharide, a disaccharide or a trisaccharide;

[0030] x3 is H, a monosaccharide (except xylose) or a disaccharide; and

[0031] ga3 is H, monosaccharide or disaccharide.

[0032] The present disclosure provides modified saponins that may have Formula I:

[0033]

[0034] in:

[0035] q1 is H or OH;

[0036] q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a component of an acetal group or an imine group;

[0037] ga5 is selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-;

[0038] Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O0-1 (CY12) 0-20 ] 0-20 A linear chain of

[0039] wherein X1 and Y1 are each independently H or a halogen atom;

[0040] Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ;

[0041] Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl;

[0042] wherein Z1 and L1 are each independently H or a halogen atom; and

[0043] Wherein Ar2 is a substituted or unsubstituted aromatic group.

[0044] The present disclosure provides a modified saponin, which can be selected from the group consisting of formulas 2D to 2H:

[0045]

[0046] The present disclosure provides pharmaceutical compositions comprising a therapeutically effective amount of a modified saponin having the formula:

[0047]

[0048] in:

[0049] q1 is H or OH;

[0050] q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a component of an acetal group or an imine group;

[0051] f3 and f4 are each independently OH or acetyl or C3 and C4 of a fucosyl unit, wherein f3 and f4 may form a cyclic ketal ring or a cyclic carbonate;

[0052] f5 is selected from H, a methyl group, a -CH2OH group, R4-NR5-C(O)-, R4-OC(O)- or R4-O-CH2-;

[0053] wherein R4 and R5 are each independently a compound having the structure R6[(CX2) 0-20 O 0-1 (CY2) 0-20 ] 0-20 A linear chain of

[0054] wherein X and Y are each independently H or a halogen atom;

[0055] wherein R6 is COOR7, C(O)NR7R8, NR7R8 or OR7;

[0056] Where R7 is Ar[(CZ2) 0-20 O 0-1 (CL2) 0-20 ] 0-20 , wherein R8 is H or alkyl;

[0057] wherein Z and L are each independently H or a halogen atom;

[0058] wherein Ar is a substituted or unsubstituted aromatic system;

[0059] ga5 is selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-;

[0060] Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 A linear chain of

[0061] wherein X1 and Y1 are each independently H or a halogen atom;

[0062] Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ;

[0063] Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl;

[0064] Wherein Z1 and L1 are each independently H or a halogen atom;

[0065] wherein Ar2 is a substituted or unsubstituted aromatic group;

[0066] r3 is H, a monosaccharide, a disaccharide or a trisaccharide;

[0067] x3 is H, a monosaccharide (except xylose) or a disaccharide; and

[0068] ga3 is H, monosaccharide or disaccharide.

[0069] The present disclosure provides pharmaceutical compositions comprising a therapeutically effective amount of a modified saponin having Formula I to treat a condition:

[0070]

[0071] in:

[0072] q1 is H or OH;

[0073] q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a component of an acetal group or an imine group;

[0074] ga5 is selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-;

[0075] Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 A linear chain of

[0076] wherein X1 and Y1 are each independently H or a halogen atom;

[0077] Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ;

[0078] Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R18 is H or alkyl;

[0079] wherein Z1 and L1 are each independently H or a halogen atom; and

[0080] Wherein Ar2 is a substituted or unsubstituted aromatic group.

[0081] The present disclosure provides a therapeutically effective amount of a modified saponin having one of Formulae 2D to 2H to treat a disorder:

[0082]

[0083] The present disclosure provides pharmaceutical compositions (such as those described above), which may also include at least one immunogen, a pharmaceutically acceptable carrier, at least one therapeutic agent (eg, at least one cancer therapeutic agent). The pharmaceutical compositions may be formulated for administration to an animal or human subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0084] Other aspects of the present disclosure will be more readily understood when the detailed description of various embodiments of the present disclosure described below is reviewed in conjunction with the accompanying drawings.

[0085] Figure 1 The preparation of MS derivatives 1D to 1H from MS II is shown.

[0086] Figure 2 The synthesis of side chains to be incorporated into 1D to 1H is shown.

[0087] Figure 3 The preparation of MS derivatives 2D to 2H from MS II is shown.

[0088] Figure 4 Derivatives 2D to 2H of the present disclosure are shown.

[0089] Figures 5A to 5C Serum IgG, IgG1 and IgG2a anti-OVA responses in mice immunized by subcutaneous route with ovalbumin (OVA) alone, or with QS-21 or VSA-2, or with one of the saponin adjuvants 1D to 1H are shown. Statistical significance was assessed by t-test (using unpaired, nonparametric and Mann-Whiteny test). *P<0.05, **P<0.01.

[0090] Figures 6A to 6C Shown are serum IgG, IgG1 and IgG2a anti-OVA responses in mice immunized by subcutaneous route with ovalbumin (OVA) alone, or with QS-21 or VSA-2, or with one of the saponin adjuvants 2D to 2H. DETAILED DESCRIPTION

[0091] The present disclosure is not limited to the particular embodiments described, and as such may, of course, vary.The terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.

[0092] When a range of values ​​is provided, each intervening value between the upper and lower limits of the range (one-tenth of the unit of the lower limit unless the context clearly indicates otherwise) and any other value or intervening value in the stated range are encompassed within the disclosure. The upper and lower limits of these smaller ranges may independently be included in smaller ranges and are also encompassed within the disclosure, subject to any explicitly excluded limits in the stated ranges. Where the stated range includes one or both of the limits, ranges excluding either or both of the included limits are also encompassed within the disclosure.

[0093] Unless otherwise indicated, the embodiments of the present disclosure will employ techniques of medicine, organic chemistry, biochemistry, molecular biology, pharmacology, etc., which are within the skill of the art and are fully explained in the literature.

[0094] The following examples are presented to provide a person of ordinary skill in the art with a complete disclosure and description of how to perform methods and use the compositions and compounds disclosed and claimed herein. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperatures, etc.), but some errors and deviations should be considered. Unless otherwise indicated, parts are parts by weight, temperatures are in ° C, and pressures are at or near atmospheric pressure. Standard temperature and pressure are defined as 20 ° C and 1 atmosphere.

[0095] Before describing the embodiments of the present disclosure in detail, it should be understood that, unless otherwise indicated, the present disclosure is not limited to specific materials, reagents, reaction materials, manufacturing processes, sizes, frequency ranges, applications, etc., because these can change. It should also be understood that the terms used herein are only for the purpose of describing specific embodiments and are not intended to be restrictive. In the present disclosure, steps may also be performed in different orders where logically possible. The embodiments of the present disclosure may also be applied to additional embodiments involving measurements beyond the examples described herein, which are not intended to be restrictive. In addition, the embodiments of the present disclosure may be combined or integrated with other measurement techniques beyond the examples described herein, which are not intended to be restrictive.

[0096] It should be noted that, as used in the specification and the appended claims, the singular forms "a and 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, reference will be made to a number of terms, which will be defined to have the following meanings, unless a contrary intention is apparent.

[0097] Each of the applications and patents cited in this text, and each document or reference cited in each of the applications and patents (including during the prosecution of each issued patent; "application cited documents"), and one of the PCT and foreign applications or patents corresponding to and / or claiming priority from any of these applications and patents, and each of the documents cited or referenced in each of the application cited documents, are hereby expressly incorporated herein by reference. In addition, documents or references cited in this text, in the reference list preceding the claims, or in the text itself; each of these documents or references ("herein cited references"), and each document or reference cited in each of the references cited herein (including any manufacturer's specifications, instructions, etc.) are hereby expressly incorporated herein by reference.

[0098] Before describing various embodiments, the following definitions are provided and shall apply unless otherwise indicated.

[0099] definition

[0100] It should be understood that "substitution" or "substituted" includes the implicit precondition that such substitution is consistent with the allowed valencies of the substituted atom and the substituent and that the substitution results in a stable compound, ie, a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or the like.

[0101] Those skilled in the art will appreciate that, if appropriate, the substituted part itself may also be substituted. For example, the substituent of the substituted alkyl may include halogen, hydroxyl, nitro, thiol, amino, azido, imino, amide, phosphoryl (including phosphonate and phosphinate), sulfonyl (including sulfate, sulfonamido, sulfamoyl and sulfonate) and silyl, as well as ether, alkylthio, carbonyl (including ketone, aldehyde, carboxylate and ester), -CF , -CN, etc. Cycloalkyl may be substituted in the same manner.

[0102] The term "acyl" as used herein, alone or in combination, refers to a carbonyl or thiocarbonyl group bonded to a radical selected from, for example, an optionally substituted hydrogen radical, an alkyl (e.g., a haloalkyl), an alkenyl, an alkynyl, an alkoxy radical ("acyloxy" includes acetoxy, butyryloxy, isovaleryloxy, phenylacetyloxy, benzoyloxy, p-methoxybenzoyloxy, and substituted acyloxy radicals such as alkoxyalkyl and haloalkoxy), an aryl, halo, heterocyclyl, heteroaryl, sulfonyl (e.g., allylsulfinylalkyl), sulfonyl (e.g., alkylsulfonylalkyl), cycloalkyl, cycloalkenyl, thioalkyl, thioaryl, amino (e.g., alkylamino or dialkylamino), and aralkyloxy radicals. Illustrative examples of "acyl" radicals are formyl, acetyl, 2-chloroacetyl, 2-bromoacetyl, benzoyl, trifluoroacetyl, phthaloyl, malonyl, nicotinoyl, and the like. As used herein, the term "acyl" refers to the group -C(O)R 26 , where 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, benzylcarbonyl and the like.

[0103] As used herein, the term "adjuvant molecule" refers to a surface protein that is capable of eliciting an immune response in a host. In a specific embodiment, the adjuvant molecule is a "membrane-anchored form" of the adjuvant molecule, which indicates that the adjuvant molecule has been engineered to include a signal peptide (SP) and a membrane anchoring sequence to direct the transport and membrane orientation of the protein. Thus, in an embodiment, the membrane-anchored form of the adjuvant molecule is a recombinant protein that includes a portion of the protein fused to the SP and the membrane anchoring sequence.

[0104] As used herein, the terms "administering" and "administration" refer to the introduction of a composition of the present disclosure (e.g., a vaccine, adjuvant, or immunogenic composition) into a subject. As used herein, "administration" may refer to oral, topical, intravenous, subcutaneous, transdermal, transdermal, intramuscular, intraarticular, parenteral, intraarteriole, intradermal, intraventricular, intraosseous, intraocular, intracranial, intraperitoneal, intralesional, intranasal, intracardiac, intraarticular, intracavernous sinus, intrathecal, intravitreal, intracerebral and intraventricular, intratympanic, intracochlear, rectal, vaginal, by inhalation, by catheter, stent, or via implanted reservoir or other device, the composition is actively or passively administered (e.g., by diffusion) to the perivascular space and adventitia. The preferred route of administering the vaccine composition is intravenous.

[0105] As used herein, the term "alkoxy" or "alkoxyalkyl" refers to an alkyl-O- group, wherein the alkyl group is as previously described. As used herein, the term "alkoxy" may refer to a C1-20 The term "oxy-hydrocarbon" refers to any of a linear, branched or cyclic, saturated or unsaturated oxy-hydrocarbon chain, including, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy and pentoxy.

[0106] As used herein, the term "alkyl", whether alone or in other terms (such as "thioalkyl" and "arylalkyl"), refers to a monovalent saturated hydrocarbon radical, which can be a straight chain (i.e., a linear chain) or a branched chain. The alkyl radicals used in the present disclosure generally contain about 1 to 20 carbon atoms, particularly about 1 to 10, 1 to 8, or 1 to 7, more particularly about 1 to 6 carbon atoms or 3 to 6. Exemplary alkyl groups include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, isopentyl, pentyl, 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, nonadecanyl, eicosyl, docosyl, n-tetracosyl, the like, and branched variants thereof. In certain aspects of the present disclosure, the alkyl radical is a C1-C6 low alkyl group, which includes or is selected from the group consisting of: methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, isopentyl, pentyl, tributyl, sec-butyl, tert-butyl, tert-pentyl and n-hexyl. The alkyl radical may optionally be substituted by a substituent 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 efficacy of the compounds. In some aspects of the present disclosure, the alkyl radical is replaced by one to five substituents, and the one to five substituents include halo, low alkoxy, low aliphatic, substituted low aliphatic, hydroxyl, cyano, nitro, thio, amino, keto, aldehyde, ester, amide, substituted amino, carboxyl, sulfonyl, sulfonyl, sulfinyl, sulfate, sulfoxide, substituted carboxyl, halogenated low alkyl (e.g., CF3), halogenated low alkoxy, hydroxycarbonyl, low alkoxycarbonyl, low alkyl carbonyl oxygen base, low alkyl carbonyl amino, alicyclic, substituted alicyclic or aryl (e.g., phenylmethylbenzyl)), heteroaryl (e.g., pyridyl) and heterocyclic (e.g., piperidyl, morpholinyl). The substituents on the alkyl group themselves can be substituted.

[0107] Ar (e.g., Ar1, Ar2, etc.) groups are aromatic systems or groups, such as aryl groups. As used herein, "aryl" refers to C5-C 20In one aspect, "aryl" can include 5-, 6-, 7-, 8-, 9- and 10-membered monocyclic aromatic groups, which can include zero to four heteroatoms, for example, functional groups corresponding to benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine and pyrimidine, etc. Those aryl groups with heteroatoms in the ring structure can also be referred to as "aryl heterocycles" or "heteroaromatic groups". The aromatic ring may be substituted at one or more ring positions with one or more substituents including, but not limited to, halogen, azido, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxy, alkoxy, amino (or quaternized amino), nitro, sulfhydryl, imino, amide, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclic, aromatic or heteroaromatic moiety, -CF3, -CN; and combinations thereof.

[0108] The term "aryl" also includes polycyclic ring systems (C5-C 30), wherein two or more carbons are common to two adjacent rings (i.e., "fused rings"), wherein at least one of the rings is an aromatic group, for example, the other one or more cyclic rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aryl and / or heterocyclic groups. Examples of heterocyclic rings include, but are not limited to, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzophenylthio, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benzotetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH carbazolyl, carbolinyl, chromanyl, benzopyranyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofurfural [2,3,b]tetrahydrofuran, furanyl ... oxazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenine, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinyl, isobenzofuranyl, isobenzodihydrofuranyl, isoindazolyl, isoindolyl, 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, phenanthroline, phenazinyl, phenothiazinyl, phenoxathiyl, phenoxazine, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidine The term "aryl" refers to any of the following groups: 1,2,3,4-thiazolyl, 1,2,5-thiadiazinyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, phenylthio and xanthenyl. One or more of the rings may be substituted as defined above for "aryl".

[0109] As used herein, the term "antibody" refers to polyclonal and monoclonal antibody preparations as well as preparations including hybrid antibodies, altered 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.

[0110] As used herein, the term "antibody" also refers to an immunoglobulin that specifically binds to the specific spatial and polar organization of another molecule and is defined as complementary to the specific spatial and polar organization of another molecule. Antibodies can be monoclonal, polyclonal or recombinant antibodies, and can be prepared by techniques well known in the art, such as immunizing a host and collecting serum (polyclonal) or by preparing a continuous hybrid cell line and collecting secreted proteins (monoclonal) or by cloning and expressing nucleotide sequences or their mutagenic forms, at least encoding the amino acid sequence required for the specific binding of natural antibodies. Antibodies can include complete immunoglobulins or fragments thereof, which include various classes and isotypes, such as IgA, IgD, IgE, IgG1, IgG2a, IgG2b and IgG3, IgM, IgY, etc. Its fragments can include Fab, Fv and F(ab')2, Fab', scFv, etc. In addition, aggregates, polymers and conjugates of immunoglobulins or their fragments can be used where appropriate, as long as the binding affinity to a specific molecule is maintained.

[0111] As used herein, the term "antigen" refers to a molecule having one or more epitopes that stimulate the host's immune system to produce a secretory, humoral and / or cellular antigen-specific response, or a DNA molecule capable of producing such an antigen in a vertebrate. The term may also be used interchangeably with "immunogen." For example, a specific antigen may 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 to trigger a host immune response.

[0112] As used herein, the term "antigenic component" refers to a component derived from an organism that can stimulate an immune response in an animal, preferably a mammal (including mice and humans). The antigenic component can be an immunogenic agent. The antigenic component can include subcellular components, including organelles, membranes, proteins, lipids, glycoproteins, and other components derived from an organism. The antigenic component can be derived from a whole organism, for example, a whole parasite, or a part of an organism, for example, a cell or tissue of an organism. In addition, a subset of proteins can be purified and recombined, for example, by size fractionation or affinity purification.

[0113] As used herein, the terms "sugar" and "saccharide" refer to polyhydroxy aldehydes, polyhydroxy ketones and their derivatives. The term includes monosaccharides such as erythrose, arabinose, allose, altrose, glucose, mannose, threose, xylose, gulose, idose, galactose, talose, aldohexose, fructose, ketohexose, ribose and aldopentose. The term also includes carbohydrates composed of monosaccharide units, including disaccharides, oligosaccharides or polysaccharides. Examples of disaccharides are sucrose, lactose and maltose. Oligosaccharides typically contain 3 to 9 monosaccharide units, while 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 the embodiments of the present disclosure where the carbohydrate is a hexose, the hexose is glucose, galactose or mannose, or a substituted hexose 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 pentoses include arabinose, fucose and ribose. The sugar residue can be connected to the compound of the present disclosure from a 1,1 bond, a 1,2 bond, a 1,3 bond, a 1,4 bond, a 1,5 bond or a 1,6 bond. The connection can be via an oxygen atom of a compound of the present disclosure. The oxygen atom can be substituted one or more times by a --CH2- or --S-- group.

[0114] The term "carboxyl" as used herein, alone or in combination, refers to -C(O)OR 25 -OR-C(-O)OR 25 , where R 25 is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, amino, thiol, aryl, heteroaryl, thioalkyl, thioaryl, thioalkoxy, heteroaryl or heterocycle, which may be optionally substituted. In aspects of the present disclosure, the carboxyl group is in esterified form and may contain a lower alkyl group as an esterifying group. In specific aspects of the present disclosure, -C(O)OR 25Provide ester or amino acid derivative.Esterified form is also specifically referred to as "carboxylate" in this article.In aspects of the present disclosure, "carboxyl" can be substituted, particularly substituted by allyl, which is optionally substituted by one or more of amino, amine, halo, alkylamino, aryl, carboxyl or heterocycle.The example of carboxyl group is methoxycarbonyl, butoxycarbonyl, tert-alkoxycarbonyl (such as tert-butoxycarbonyl), arylmethoxycarbonyl with one or two aryl radicals including but not limited to phenyl optionally substituted by, for example, lower alkyl, lower alkoxy, hydroxyl, halo and / or nitro, such as benzyloxycarbonyl, methoxybenzyloxycarbonyl, diphenylmethoxycarbonyl, 2-bromoethoxycarbonyl, 2-iodoethoxycarbonyl tert-butylcarbonyl, 4-nitrobenzyloxycarbonyl, diphenylmethoxycarbonyl, phenylhydroxycarbonyl, di-(4-methoxyphenylmethoxycarbonyl, 2-bromoethoxycarbonyl, 2-iodoethoxycarbonyl, 2-trimethylsilane The carboxyl group of esterified form is siloxycarbonyl group, including organosiloxycarbonyl. The silicon substituent in this compound can be replaced by low alkyl (e.g., methyl), alkoxy (e.g., methoxy) and / or halo (e.g., chlorine). The example of silicon substituent includes trimethylsilyl and dimethyl tert-butylsilyl. In the aspect of the present disclosure, the carboxyl group can be alkoxycarbonyl, particularly methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, tert-butoxycarbonyl, tert-pentyloxycarbonyl, heptyloxycarbonyl, especially methoxycarbonyl or ethoxycarbonyl.

[0115] The term "imine" refers to R'1-N=CR"R"', wherein R', R" and R"' are each independently selected from an alkyl, alkenyl, carbocyclic, heterocyclic, aryl or heteroaryl group.

[0116] As used herein, the term "composition" refers to a product comprising specified ingredients in specified amounts, as well as any product resulting directly or indirectly from the combination of specified ingredients in specified amounts. This term in relation to pharmaceutical compositions is intended to encompass products comprising active ingredients and inert ingredients constituting carriers, as well as any product resulting directly or indirectly from the combination, complexation or aggregation of any two or more ingredients, or from the dissociation of one or more ingredients, or from other types of reactions or interactions of one or more ingredients. Thus, the pharmaceutical compositions of the present disclosure encompass any composition prepared by mixing a compound of the present disclosure and a pharmaceutically acceptable carrier.

[0117] 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 considered. Therefore, pharmaceutical compositions of the present disclosure include those containing 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 can vary and will depend on the effective dose of each ingredient. Generally, respective effective doses will be used. Therefore, for example, but not intended to be limiting, when the compounds of the present disclosure are combined with another medicament, the weight ratio of the compounds of the present disclosure to the other medicament 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 with other active ingredients is also generally within the above range, but in each case, the effective dose of each active ingredient should be used. In such combinations, the compounds of the present disclosure and other active agents can be administered alone or in combination. In addition, the administration of one ingredient can be before, at the same time, or after the administration of other medicaments.

[0118] The compositions of the present disclosure can be liquid solutions, suspensions, emulsions, tablets, pills, capsules, sustained release formulations or powders. The compositions can be formulated into suppositories together with conventional binders and carriers such as triglycerides. Oral formulations can include standard carriers such as pharmaceutical grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc. Various delivery systems are known and can be used to administer the compositions of the present disclosure, for example, encapsulated in liposomes, microparticles, microcapsules, etc.

[0119] The therapeutic composition of the present disclosure may include a carrier, such as one or more of a polymer, a carbohydrate, a peptide or a derivative thereof, which may be covalently attached to the compound directly or indirectly. The carrier may be substituted by a substituent as described herein, including but not limited to one or more alkyl, amino, nitro, halogen, thiol, alkylthio, sulfate, sulfonyl, sulfinyl, sulfoxide, hydroxyl group. In aspects of the present disclosure, the carrier is an amino acid, including alanine, glycine, proline, methionine, serine, threonine, asparagine, alanyl-alanyl, prolyl-methionyl or glycyl-glycyl. The carrier may also include a molecule that targets the compound of the present disclosure to a specific tissue or organ.

[0120] In view of this knowledge and the disclosure of the present application (including examples), the compounds of the present disclosure can be prepared using reactions and methods generally known to those of ordinary skill in the art. The reaction is carried out in a solvent that is compatible with the reagents and materials used and is suitable for realizing these reactions. It should be understood by those skilled in the art of organic synthesis that the functional groups present on the compound should be consistent with the proposed reaction steps. Sometimes, in order to obtain the desired compounds of the present disclosure, it is necessary to adjust the order of the synthesis steps, or select a specific process scheme rather than another. It should also be recognized that when developing synthetic routes, another major consideration is the selection of protective groups for protecting the reactive functional groups present in the compounds described in the present disclosure. An authoritative description of many alternatives describing technical personnel is Greene and Wuts (Protective Groups In Organic Synthesis, Wiley and Sons, 1991).

[0121] Compounds of the present disclosure can be formulated into pharmaceutical compositions for application to subjects by appropriate methods known in the art. Pharmaceutical compositions of the present disclosure or parts thereof include suitable pharmaceutically acceptable carriers, excipients and carriers selected based on the expected form of administration and consistent with conventional drug practice. Suitable pharmaceutical carriers, excipients and carriers are described in standard text Remington: The Science and Practice of Pharmacy (21st edition 2005, University of the Sciences in Philadelphia (editor), Mack Publishing Company) and in the United States Pharmacopeia: National Formulary (The United States Pharmacopeia: The National Formulary) (USP 24NF19) (published in 1999). For example, for oral administration in the form of capsules or tablets, active ingredients can be combined with oral, nontoxic pharmaceutically acceptable inert carriers (such as lactose, starch, sucrose, methylcellulose, magnesium stearate, glucose, calcium, sulfate, dicalcium phosphate, mannitol, sorbitol, etc.). For oral administration in liquid form, the tablet component can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier (such as ethanol, glycerol, water, etc.). Suitable binders (e.g., gelatin, starch, corn sweeteners, natural sugars (including glucose); natural and synthetic gums and waxes), lubricants (e.g., sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate and sodium chloride), disintegrants (e.g., starch, methylcellulose, agar, bentonite and xanthan gum), flavoring agents and coloring agents can also be combined in the composition or its components. The composition as described herein may further contain a wetting agent or emulsifier or a pH buffer.

[0122] As used herein, the term "immunogenic compositions" are those that, when injected into a host, result in the production of specific antibodies or cellular immunity.

[0123] The immunogenic compositions and / or vaccines disclosed herein can be formulated by any of the methods known in the art. They can generally be prepared as injections or formulations for intranasal administration, whether liquid solutions or suspensions. Solid forms suitable for being dissolved or suspended in liquids prior to injection or other administration can also be prepared. The formulations can also be, for example, emulsified, or the protein / peptide can be encapsulated in liposomes.

[0124] The active immunogenic component is usually mixed with a pharmaceutically acceptable excipient or carrier that is compatible with the active ingredient. Suitable excipients include, but are not limited to, water, saline, dextrose, glycerol, ethanol, and the like, and combinations thereof. The concentration of the immunogenic polypeptide in the injectable, aerosol, or nasal formulation is usually in the range of about 0.2 to 5 mg / ml. Similar doses may be applied to other mucosal surfaces.

[0125] In addition, if necessary, the vaccine may contain a small amount of auxiliary substances, such as wetting agents or emulsifiers, pH buffers and / or other agents, which enhance the effectiveness of the vaccine. Examples of potentially effective agents include, but are not limited to, aluminum hydroxide; N-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP); N-acetyl-normal-muramyl-L-alanyl-D-isoglutamine (CGP 11637, referred to as nor-MDP); N-acetylmuramyl-L-alanyl-D-isoglutamido-L-alanine-2-(1'-2'-dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)-ethylamine (CGP 19835A, referred to as MTP-PE); and RIBI, which contains three components extracted from bacteria: monophosphoryl lipid A, trehalose dimycolate and cell wall skeleton (MPL+TDM+CWS) in 2% squalene / Tween 80 emulsion. The effectiveness of an adjuvant can be determined by measuring the amount of antibodies (particularly IgG, IgM or IgA) against the immunogen produced by administration of the immunogen in a vaccine comprising the adjuvant in question.Alternative formulations and modes of administration may also be used.

[0126] As is known in the art, the immunogenic compositions and / or vaccines of the present disclosure can be administered in a manner compatible with the dosage formulation and in an amount and manner that is effective for prevention and / or treatment. The amount to be administered is typically in the range of about 1 to 1,000 micrograms of viral surface envelope glycoproteins and / or adjuvant molecules per dose, more typically in the range of about 5 to 500 micrograms of glycoproteins and / or adjuvant molecules per dose, depending on the nature of the antigen and / or adjuvant molecules undergoing treatment, the ability of the host's immune system to synthesize antibodies, and the degree of protection required. The precise amount of active ingredient that needs to be administered may depend on the judgment of the physician or veterinarian, and may be unique to each individual, but such determination is within the skill of such practitioners.

[0127] The vaccine or immunogenic composition can be administered in a single dose; a two-dose regimen, e.g., two to eight weeks apart; or a multiple-dose regimen. A multiple-dose regimen is a regimen in which the primary vaccination course may include 1 to 10 or more individual doses, followed by other doses administered at subsequent time intervals as needed to maintain and / or enhance the immune response (e.g., a second dose is administered 1 to 4 months apart, and subsequent doses are administered several months later if necessary).

[0128] As used herein, the term "immunogenic fragment" refers to a fragment of an immunogen that includes one or more epitopes and can therefore modulate an immune response or can serve as an adjuvant for a co-administered antigen. Such fragments can be identified using any number of epitope mapping techniques well known in the art (see, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66 (Morris, GE, ed., 1996) Humana Press, Totowa, NJ).

[0129] The length of an immunogenic fragment can be at least about 2 amino acids, 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 strict upper limit to the length of the fragment, which can include almost the entire length of the protein sequence or even a fusion protein containing two or more epitopes.

[0130] 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 divided into five categories: IgM, IgG, IgA, IgD and IgE. IgG is the most abundant antibody class in the body and is assumed to be a distorted "Y" shaped configuration. Except for IgM, immunoglobulins are composed of four peptide chains, which are connected by intrachain and interchain disulfide bonds. IgG consists of two polypeptide heavy chains (H chains) and two polypeptide light chains (L chains) coupled by non-covalent disulfide bonds.

[0131] As used herein, the term "immunological response" refers to the production of a humoral and / or cellular immune response in a subject against an antigen present in a composition of interest. For purposes of this disclosure, a "humoral immune response" refers to an immune response mediated by antibody molecules, while a "cellular immune response" is an immune response mediated by T lymphocytes and / or other white blood cells.

[0132] One aspect of cellular immunity involves the antigen-specific response of cytolytic T cells ("CTL"). CTLs are specific for peptide antigens that are associated with proteins encoded by the major histocompatibility complex (MHC) and expressed on the cell surface. CTLs help induce and promote the destruction of intracellular microorganisms or the lysis of cells infected by such microorganisms. Another aspect of cellular immunity involves the antigen-specific response of helper T cells. The role of helper T cells is to help stimulate the function of nonspecific effector cells and focus their activity on cells that display peptide antigens associated with MHC molecules on their surfaces. "Cellular immune response" also refers to the production of cytokines, chemokines and other such molecules produced by activated T cells and / or other white blood cells (including those derived from CD4+ and CD8+ T cells). Therefore, the immunological response may include one or more of the following effects: B cells produce antibodies; and / or the activation of suppressive T cells and / or γδ T cells specifically for one or more antigens present in the composition or vaccine of interest. These responses can be used to neutralize infectivity and / or mediate antibody complement or antibody-dependent cellular cytotoxicity (ADCC), thereby providing protection for the immune host. Such responses can be determined using standard immunoassays and neutralization assays well known in the art.

[0133] As used herein, the term "immunogenic amount" refers to an amount that is capable of eliciting the production of antibodies against a virus in a host to which the vaccine has been administered.

[0134] As used herein, the term "immunogenic carrier" refers to a composition that enhances the immunogenicity of virions from any of the viruses discussed herein. Such carriers include, but are not limited to, proteins and polysaccharides and microspheres formulated using, for example, biodegradable polymers (such as DL-lactide-glycolide), liposomes, and bacterial cells and membranes. Protein carriers can be connected to proteases or peptides derived therefrom to form fusion proteins by recombinant or synthetic techniques or by chemical coupling. Useful carriers and ways to couple such carriers to polypeptide antigens are known in the art.

[0135] As used herein, the term "immunoenhancer" means a substance that, when mixed with an immunogen, induces a stronger immune response than the immunogen alone. For example, an immunoenhancer can enhance immunogenicity and provide a superior immune response. An immunoenhancer can, for example, work by enhancing the expression of co-stimulators on macrophages and other antigen presenting cells.

[0136] As used herein, the terms "subject," "individual," or "patient" are used interchangeably and refer to an animal, preferably a warm-blooded animal, such as a mammal. Mammals include, but are not limited to, any member of the class Mammalia. The mammals that are subjects or patients in the present disclosure may be from the orders Primates, Carnivora, Proboscidea, Perissodactyla, Artiodactyla, Rodentia, and Lagomorpha. In specific embodiments, the mammal is a human. In other embodiments, animals may be treated; these animals may be vertebrates, including both birds and mammals. In aspects of the present disclosure, the term includes livestock raised for food or as pets, including equines, bovines, sheep, poultry, fish, pigs, canines, felines, and zoo animals, goats, apes (e.g., gorillas or chimpanzees), and rodents, such as rats and mice.

[0137] As used herein, the term "pharmaceutically acceptable carrier" refers to a diluent, adjuvant, excipient or carrier that is used with the probe of the present disclosure and approved by a federal or state government regulatory agency or listed in the U.S. Pharmacopeia or other recognized pharmacopoeia for use in animals and more specifically in humans. Such pharmaceutical carriers can be liquids, such as water and oils, including oils of petroleum, animal, plant or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Pharmaceutical carriers can be salts, gum arabic, gelatin, starch paste, talc, keratin, colloidal silicon dioxide, urea, etc. When applied to a patient, the probe and pharmaceutically acceptable carrier can be sterile. When the probe is administered intravenously, water is a useful carrier. Saline solutions and aqueous glucose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical carriers also include excipients, such as glucose, lactose, sucrose, glyceryl monostearate, sodium chloride, glycerol, propylene glycol, water, ethanol, etc. If desired, the composition may also contain a small amount of a wetting or emulsifying agent or a pH buffering agent. The composition of the present invention may advantageously take the form of a solution, emulsion, sustained release formulation or any other form suitable for use.

[0138] As used herein, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0139] As used herein, the term "vaccine" refers to an immunogenic amount of one or more virions, fragments or subunits thereof. Such vaccines may include one or more viral surface envelope glycoproteins and portions thereof on the surface of the virion and adjuvant molecules and portions thereof, or in combination with another protein or other immunogen, such as one or more additional viral components naturally associated with the viral particle or an epitope peptide derived therefrom.

[0140] As used herein, the terms "effective amount" and "amount effective" refer to an amount sufficient to achieve a desired result, or to act on the prevention of an undesirable condition or disease or condition, or to enhance and / or modulate the subject's immune system to a desired response to certain pathogens (e.g., viruses). For example, a "therapeutically effective amount" refers to an amount sufficient to achieve a desired therapeutic result, or to act on the prevention of an undesirable symptom or disease or condition, and / or to modulate the subject's immune system to a desired response to certain pathogens, but generally insufficient to cause adverse side effects. The specific therapeutically effective dosage level for any particular patient will depend on a variety of factors, including: the specific composition employed; the patient's age, weight, general health, sex, and diet; the time of administration; the route of administration; the excretion rate of the specific compound employed; the duration of treatment; drugs used in combination or concurrently with the specific compound employed, as well as similar factors well known in the medical field. For example, it is within the skill of the art to start the dose of the compound at a level lower than that required to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved. If necessary, the effective daily dose can be divided into multiple doses for the purpose of administration. Therefore, a single dose composition may contain such amounts or submultiples thereof to constitute the daily dose. In the case of any contraindications, the dosage may be adjusted by the individual physician. The dosage may vary and may be administered in one or more doses per day for one or more days. Guidance on appropriate dosages for a given class of drug products can be found in the literature.

[0141] As used herein, the terms "treating" and "treatment" may generally refer to obtaining a desired pharmacological and / or physiological effect. The effect may be, but need not necessarily be, preventive in terms of preventing or partially preventing a disease, its symptoms or conditions (such as infection and its consequences) and / or adjusting the subject's immune system to a desired response to certain pathogens. The effect may be therapeutic, i.e., partially or completely curing a disease, condition, symptom, or side effects attributed to the disease, illness, or condition. As used herein, the term "treatment" may include any treatment of an infection in a subject, particularly a human, and may include any one or more of the following: (a) preventing the occurrence of the disease or infection in a subject who may be susceptible to the disease or infection but has not yet been diagnosed with the disease or infection; (b) inhibiting the disease or infection, i.e., preventing its development; and (c) alleviating the disease or infection, i.e., alleviating or ameliorating the disease and / or its symptoms or conditions, (d) and / or adjusting the subject's immune system to a desired response to certain pathogens. As used herein, the term "treatment" may refer to a single therapeutic treatment, a single preventive treatment, or both a therapeutic treatment and a preventive treatment. Those in need of treatment (subjects in need thereof) may include those already suffering from the condition and / or those in which the condition is to be prevented. As used herein, the term "treatment" may include inhibiting a disease, illness, or condition, such as preventing its progression; and alleviating a disease, illness, or condition, such as causing the disease, illness, and / or condition to subside, and / or adjusting the subject's immune system to a desired response to certain pathogens. Treating a disease, illness, or condition may include improving at least one symptom of a particular disease, illness, or condition, even if the underlying pathophysiology is not affected, for example, such as treating pain in a subject by administering an analgesic, even if such a drug cannot treat the cause of the pain.

[0142] As used herein, "therapeutic" may refer to treating, curing and / or ameliorating a disease, disorder, condition or side effect, or to reducing the rate of progression of a disease, disorder, condition or side effect and / or modulating a subject's immune system to a desired response against certain pathogens.

[0143] The term "pharmaceutically acceptable" describes a material that is not biologically or otherwise undesirable, ie, does not induce unacceptable levels of undesirable biological effects or interact in a deleterious manner.

[0144] The term "pharmaceutically acceptable prodrug" or "prodrug" means those prodrugs of the compounds of the present disclosure that are, within the scope of sound medical judgment, suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, etc., 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 to the parent compound having the structure of the disclosed compound, for example, by hydrolysis in the blood. A comprehensive discussion is provided in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, ACS Symposium Series, Vol. 14, and Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press (1987).

[0145] In 1974, Dalsgaard et al. first described saponin preparations isolated from the South American soap tree as having adjuvant activity ("Saponin adjuvants", Archiv. fur die gesamte Virusforschung, Vol. 44, Springer Verlag, Berlin, pp. 243-254). Purified fragments of Quil A have been separated by HPLC, which retain 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 the soap tree that can induce CD8+ cytotoxic T cells (CTL), Th1 cells and a major IgG2a antibody response.

[0146] The saponins of the present disclosure may be used in an amount of between 1 and 100 μg per human dose of the adjuvant composition (at a level of about 50 μg, e.g., between 40 and 60 μg, suitably between 45 and 55 μg or between 49 and 51 μg or 50 μg). In some embodiments, a human dose of the adjuvant composition may comprise QS21 at a level of about 25 μg, e.g., between 20 and 30 μg, suitably between 21 and 29 μg or between 22 and 28 μg or between 28 and 27 μg or between 24 and 26 μg or 25 μg.

[0147] When adjuvant is combined with antigen composition in liquid form, adjuvant composition will be in the volume suitable for human dose, which is approximately half of the expected final volume of human dose.For example, the adjuvant of 500 μ l volume is used for the expected final human dose of 1 μ l, or the adjuvant of 250 μ l volume is used for the expected final human dose of 0.5 ml.When combined with antigen composition, adjuvant composition is diluted to provide the final human dose of vaccine.Certainly, the final volume of such dosage will change according to the initial volume of adjuvant composition and the volume of the antigen composition added to the adjuvant composition.In an alternative embodiment, aqueous adjuvant is used to reconstruct lyophilized antigen composition.In this embodiment, the adjuvant composition of human dose suitable volume is approximately equal to the final volume of human dose.Liquid adjuvant composition is added in the vial containing lyophilized antigen composition and is used to reconstruct the lyophilized antigen composition.

[0148] abbreviation

[0149] 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; sc, subcutaneous; ESI-TOF, electrospray ionization time-of-flight mass spectrometry; ELISA, enzyme-linked immunosorbent assay;

[0150] discuss

[0151] The present disclosure provides compounds including modified saponin compounds, pharmaceutical compositions including modified saponin compounds, methods of using the modified saponin compounds and the pharmaceutical compositions, methods of preparing modified saponin compounds, etc. The compounds and pharmaceutical compositions of the present disclosure can be used in combination with one or more other therapeutic agents for the treatment of 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 to treat viral infections.

[0152] One aspect of the present disclosure encompasses embodiments of modified saponins having the formula:

[0153]

[0154] in:

[0155] q1 can be H or OH;

[0156] q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a component of an acetal group or an imine group;

[0157] f3 and f4 are each independently OH or acetyl or C3 and C4 of a fucosyl unit, wherein f3 and f4 may form a cyclic ketal ring or a cyclic carbonate;

[0158] f5 is selected from H, a methyl group, a -CH2OH group, R4-NR5-C(O)-, R4-OC(O)- or R4-O-CH2-;

[0159] wherein R4 and R5 are each independently a compound having the structure R6[(CX2) 0-20 O 0-1 (CY2) 0-20 ] 0-20 A linear chain of

[0160] wherein X and Y are each independently H or a halogen atom;

[0161] wherein R6 is COOR7, C(O)NR7R8, NR7R8 or OR7;

[0162] Where R7 is Ar[(CZ2) 0-20 O 0-1 (CL2) 0-20 ] 0-20 , wherein R8 is H or alkyl;

[0163] wherein Z and L are each independently H or a halogen atom;

[0164] wherein Ar is a substituted or unsubstituted aromatic system;

[0165] ga5 is selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-;

[0166] Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 A linear chain of

[0167] wherein X1 and Y1 are each independently H or a halogen atom;

[0168] Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ;

[0169] Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl;

[0170] Wherein Z1 and L1 are each independently H or a halogen atom;

[0171] wherein Ar2 is a substituted or unsubstituted aromatic group;

[0172] r3 is H, a monosaccharide, a disaccharide or a trisaccharide;

[0173] x3 is H, a monosaccharide (except xylose) or a disaccharide; and

[0174] ga3 is H, monosaccharide or disaccharide.

[0175] In some embodiments, the modified saponin may have Formula I:

[0176]

[0177] in:

[0178] q1 is H or OH;

[0179] q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a component of an acetal group or an imine group;

[0180] ga5 is selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-;

[0181] Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 A linear chain of

[0182] wherein X1 and Y1 are each independently H or a halogen atom;

[0183] Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18OR 17 ;

[0184] Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl;

[0185] wherein Z1 and L1 are each independently H or a halogen atom; and

[0186] Wherein Ar2 is a substituted or unsubstituted aromatic group.

[0187] In other embodiments, the modified saponin can be selected from the group consisting of formulas 2D to 2H:

[0188]

[0189] Another aspect of the present disclosure encompasses embodiments of pharmaceutical compositions comprising modified saponins having the formula:

[0190]

[0191] in:

[0192] q1 can be H or OH;

[0193] q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a component of an acetal group or an imine group;

[0194] f3 and f4 are each independently OH or acetyl or C3 and C4 of a fucosyl unit, wherein f3 and f4 may form a cyclic ketal ring or a cyclic carbonate;

[0195] f5 is selected from H, a methyl group, a -CH2OH group, R4-NR5-C(O)-, R4-OC(O)- or R4-O-CH2-;

[0196] wherein R4 and R5 are each independently a compound having the structure R6[(CX2) 0-20 O 0-1 (CY2) 0-20 ] 0-20 A linear chain of

[0197] wherein X and Y are each independently H or a halogen atom;

[0198] wherein R6 is COOR7, C(O)NR7R8, NR7R8 or OR7;

[0199] Where R7 is Ar[(CZ2) 0-20O 0-1 (CL2) 0-20 ] 0-20 , wherein R8 is H or alkyl;

[0200] wherein Z and L are each independently H or a halogen atom;

[0201] wherein Ar is a substituted or unsubstituted aromatic system;

[0202] ga5 is selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-;

[0203] Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 A linear chain of

[0204] wherein X1 and Y1 are each independently H or a halogen atom;

[0205] Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ;

[0206] Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl;

[0207] Wherein Z1 and L1 are each independently H or a halogen atom;

[0208] wherein Ar2 is a substituted or unsubstituted aromatic group;

[0209] r3 is H, a monosaccharide, a disaccharide or a trisaccharide;

[0210] x3 is H, a monosaccharide (except xylose) or a disaccharide; and

[0211] ga3 is H, monosaccharide or disaccharide.

[0212] Another aspect of the present disclosure encompasses embodiments of pharmaceutical compositions comprising modified saponins having Formula I:

[0213]

[0214] in:

[0215] q1 is H or OH;

[0216] q2 and q3 are each independently selected from CHO, CH3, CH2OH, H, COOH, a component of an acetal group or an imine group;

[0217] ga5 is selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-;

[0218] Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 A linear chain of

[0219] wherein X1 and Y1 are each independently H or a halogen atom;

[0220] Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ;

[0221] Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl;

[0222] wherein Z1 and L1 are each independently H or a halogen atom; and

[0223] Wherein Ar2 is a substituted or unsubstituted aromatic group.

[0224] Another aspect of the present disclosure encompasses embodiments of pharmaceutical compositions comprising modified saponins having Formulae 2D to 2H:

[0225]

[0226] In some embodiments of this aspect of the disclosure, the pharmaceutical composition comprises a therapeutically effective amount of a modified saponin to treat a condition (e.g., an infection, a disease, 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), a pharmaceutical agent (e.g., a cancer agent), a pharmaceutically acceptable carrier, etc. The disclosure provides for administering a pharmaceutical composition comprising a therapeutically effective amount of a modified saponin of an active agent or a pharmaceutically acceptable salt of a modified saponin of an active agent, and a pharmaceutically acceptable carrier to a subject in need thereof.

[0227] The condition to be treated in a subject (e.g., a mammal) in need of treatment may include those conditions that the vaccine is directed against. The condition may be an infection (e.g., a pathogen infection, such as a viral infection, such as a coronavirus infection, HIV infection, influenza infection, hepatitis), a disease (such as cancer, etc.), an infection or a non-viral hyper-inflammatory / immune hyper-activation condition.

[0228] In some embodiments of this aspect of the present disclosure, the pharmaceutical composition may further comprise: at least one cancer therapeutic agent. The at least one chemotherapeutic agent and the saponin derivative may be mixed or covalently linked to each other in a pharmaceutically acceptable formulation; and a pharmaceutically acceptable carrier. The cancer therapeutic agent may be present in a therapeutically effective amount to treat the condition.

[0229] Yet another aspect of the present disclosure encompasses embodiments of methods of increasing the immunogenicity of an immunogen when administered to an animal or human subject, the method comprising the step of administering to the subject a vaccine comprising at least a pharmaceutical composition according to the present disclosure.

[0230] Yet another aspect of the present disclosure encompasses embodiments of synthetic routes for synthesizing saponin derivatives, the synthetic routes comprising coupling natural saponins with functionalized side chain molecules, wherein the functionalized side chains comprise amino groups or hydroxyl groups.

[0231] In some embodiments of this aspect of the disclosure, natural saponins can be obtained from Gac fruit.

[0232] In some embodiments of this aspect of the disclosure, natural saponins can be coupled to functionalized side chain molecules via an amide formation reaction or an ester formation reaction.

[0233] Drug formulations and routes of administration

[0234] Embodiments of the present disclosure include medicaments identified herein (e.g., modified saponins) and can be formulated with one or more pharmaceutically acceptable excipients, diluents, carriers, and / or adjuvants. In addition, embodiments of the present disclosure include medicaments formulated with one or more pharmaceutically acceptable auxiliary substances. In particular, medicaments can be formulated with one or more pharmaceutically acceptable excipients, diluents, carriers, and / or adjuvants to provide embodiments of compositions of the present disclosure.

[0235] A variety of pharmaceutically acceptable excipients are known in the art. Pharmaceutically acceptable excipients have been fully described in a variety of 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) HC Ansel et al., ed., 7th edition, Lippincott, Williams, & Wilkins; and Handbook of Pharmaceutical Excipients (2000) AK Kibbe et al., ed., 3rd edition, Amer. Pharmaceutical Assoc.

[0236] Pharmaceutically acceptable excipients, such as vehicles, adjuvants, carriers or diluents, are readily available to the public. In addition, pharmaceutically acceptable auxiliary substances, such as pH adjusters and buffers, tonicity adjusters, stabilizers, wetting agents, etc., are readily available to the public.

[0237] In embodiments of the present disclosure, the medicament can be administered to the subject in any manner that produces the desired effect. Thus, the binding agent can be incorporated into a variety of formulations for therapeutic administration. For example, the medicament can be formulated into a pharmaceutical composition by combining with an appropriate, pharmaceutically acceptable carrier or diluent, and can be formulated into a solid, semisolid, liquid or gaseous form of preparation, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants and aerosols.

[0238] In pharmaceutical dosage forms, the medicament may be administered in the form of its pharmaceutically acceptable salt, or the subject active composition may be used alone or in appropriate association and combination with other pharmaceutically active compounds. The following methods and excipients are merely exemplary and are not limiting in any way.

[0239] For oral preparations, the medicament can be used alone or in combination with appropriate additives to make tablets, powders, granules or capsules, for example, with conventional additives such as lactose, mannitol, corn starch or potato starch; with binders such as crystalline cellulose, cellulose derivatives, gum arabic, corn starch or gelatin; with disintegrants such as corn starch, potato starch or sodium carboxymethyl cellulose; with lubricants such as talc or magnesium stearate; and, if necessary, with diluents, buffers, wetting agents, preservatives and flavoring agents.

[0240] Embodiments of the medicament can be formulated into injection preparations by dissolving, suspending or emulsifying it in an aqueous or non-aqueous solvent (such as vegetable oil or other similar oils, synthetic fatty acid glycerides, higher fatty acid esters or propylene glycol); and if necessary, formulated with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers and preservatives.

[0241] Embodiments of the agents may be used in aerosol formulations for administration via inhalation. Embodiments of the agents may be formulated into pressurized acceptable propellants such as dichlorodifluoromethane, propane, nitrogen, and the like.

[0242] In addition, embodiments of the medicament can be made into suppositories by mixing with various bases such as emulsifying bases or water-soluble bases. Embodiments of the medicament can be administered rectally via suppositories. Suppositories can contain carriers such as cocoa butter, carbowax, and polyethylene glycol, which melt at body temperature but solidify at room temperature.

[0243] Unit dosage forms such as syrups, elixirs and suspensions for oral or rectal administration can be provided, wherein each dosage unit, e.g., teaspoon, tablespoon, tablet or capsule, contains a predetermined amount of a 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, saline or another pharmaceutically acceptable carrier.

[0244] According to the present disclosure, embodiments of the medicaments can be formulated in injectable compositions. Typically, injectable compositions are prepared as liquid solutions or suspensions; solid forms suitable for dissolving or suspending in a liquid carrier prior to injection can also be prepared. According to the present disclosure, the formulations can also be emulsified or the active ingredient (triamino-pyridine derivative and / or labeled triamino-pyridine derivative) can be encapsulated in a liposome carrier.

[0245] In embodiments, the medicament can be formulated for delivery by a continuous delivery system. The term "continuous delivery system" is used interchangeably herein with "controlled delivery system" and encompasses a continuous (e.g., controlled) delivery device (e.g., pump) in combination with a catheter, injection device, etc. (which are known in the art).

[0246] Mechanical or electromechanical infusion pumps may also be suitable for use with the present disclosure. Examples of such devices include those described in, for example, 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. In general, delivery of a medicament may be accomplished using any of a variety of refillable pump systems. The pump provides consistent, controlled release over time. In some embodiments, the medicament may be a liquid formulation in a drug-impermeable reservoir and delivered to an individual in a continuous manner.

[0247] In one embodiment, the drug delivery system is an at least partially implantable device. Methods and devices well known in the art can be used to implant the implantable device into any suitable implantation site. The implantation site is a site in the subject's body where the drug delivery device is introduced and positioned. The implantation site includes but is not necessarily limited to other suitable sites in the subdermal, subcutaneous, intramuscular or subject's body. In certain embodiments, a subcutaneous implantation site is used because the implantation and removal of the drug delivery device are convenient.

[0248] The drug release device suitable for use in the present disclosure can be based on any of a variety of operating modes. For example, the drug release device can be based on a diffusion system, a convection system, or an erosion system (e.g., an erosion-based system). For example, the drug release device can be an electrochemical pump, an osmotic pump, an electroosmotic pump, a vapor pressure pump, or an osmotic explosion matrix, for example, wherein the drug is incorporated into a polymer and the polymer provides release of the drug formulation as the drug-impregnated polymeric material (e.g., a biodegradable, drug-impregnated polymeric material) degrades. In other embodiments, the drug release device is based on an electrical diffusion system, an electrolytic pump, an effervescent pump, a piezoelectric pump, a hydrolysis system, etc.

[0249] Drug release devices based on mechanical or electromechanical infusion pumps may also be suitable for use with the present disclosure. Examples of such devices include those described below: for example, U.S. Patent Nos. 4,692,147; 4,360,019; 4,487,603; 4,360,019; 4,725,852, etc. In general, any one of a variety of refillable, non-replaceable pump systems can be used to complete the subject treatment method. Pumps and other convective systems are generally preferred because they are generally more consistent and controlled release over time. Osmotic pumps are used in some embodiments (see, for example, PCT published application No. WO 97 / 27840 and U.S. Patent Nos. 5,985,305 and 5,728,396) due to their more consistent controlled release and relatively small size combination advantages. Exemplary osmotically driven devices suitable for use in the present disclosure include, but are not necessarily limited to, those described in: 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;etc.

[0250] In some embodiments, the drug delivery device is an implantable device. The drug delivery device can be implanted into any suitable implantation site using methods and devices well known in the art. As noted herein, an implantation site is a site in a subject's body where a drug delivery device is introduced and positioned. Implantation sites include but are not necessarily limited to subdermal, subcutaneous, intramuscular, or other suitable sites in a subject's body.

[0251] In some embodiments, an implantable drug delivery system can be used to deliver the agent, for example, a system that can be programmed to provide for administration of the agent. Exemplary programmable implant systems include implantable infusion pumps. Exemplary implantable infusion pumps or devices used in conjunction with such pumps are described, for example, in U.S. Pat. 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. Another exemplary device that may be suitable for use in the present disclosure is the Synchromed infusion pump (Medtronic).

[0252] Suitable excipient carriers for the medicament are, for example, water, saline, dextrose, glycerol, ethanol, etc. and combinations thereof. In addition, if desired, the carrier may contain a small amount of auxiliary substances, such as a wetting agent or emulsifier or pH buffer. Methods for preparing such dosage forms are known to those skilled in the art or will be apparent after considering this disclosure. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania, 17th edition, 1985. In any case, the composition or formulation to be administered will contain an amount of the medicament sufficient to achieve the desired state in the subject being treated.

[0253] Compositions of the present disclosure may include those comprising sustained release or controlled release matrix. In addition, embodiments of the present disclosure may be used in combination with other treatments using sustained release formulations. As used herein, sustained release matrix is ​​a matrix made of a material (usually a polymer) that can be hydrolyzed by enzymes or acid groups or by dissolving and degrading. Once inserted into the body, the matrix will be subject to the effects of enzymes and body fluids. Sustained release matrix is ​​ideally selected from biocompatible materials, such as liposomes, polylactide (polylactic acid), polyglycolide (polymer of glycolic acid), polylactide co-glycolide (copolymer of lactic acid and glycolic acid), polyanhydrides, poly (original) 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, polyethylene propylene, polyvinyl pyrrolidone and silicones. Exemplary biodegradable matrixes include polylactide matrix, polyglycolide matrix and polylactide co-glycolide (copolymer of lactic acid and glycolic acid) matrix.

[0254] In another embodiment, the pharmaceutical composition of the present disclosure (and the composition of the combination) can be delivered in a controlled release system. For example, intravenous infusion, implantable osmotic pumps, transdermal patches, liposomes or other modes of administration can be used to administer the agent. In one embodiment, a pump can be used (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). In another embodiment, a polymeric material can be used. In yet another embodiment, the controlled release system is placed near the therapeutic target, so only a portion of the systemic dose is required. In yet another embodiment, the controlled release system is placed near the therapeutic target, so only a portion of the systemic dose is required. Other controlled release systems are discussed in the review of Langer (1990). Science 249: 1527-1533.

[0255] In another embodiment, the compositions of the present disclosure (and the compositions of the combinations alone or together) include those formed by impregnating the agents described herein into absorbent materials (such as sutures, bandages, and gauze), or those coated onto the surface of solid phase materials (such as surgical staples, zippers, and catheters for delivery of the compositions. Other delivery systems of this type will be apparent to those skilled in the art in light of this disclosure.

[0256] dose

[0257] Embodiments of medicaments (e.g., modified saponins) can be used to subjects with one or more dosages. Technicians will readily appreciate that dosage levels can vary with the function of the particular medicament administered, the severity of the symptoms, and the sensitivity of the subject to side effects. Those skilled in the art can easily determine the preferred dosage of a given compound in a variety of ways.

[0258] In an embodiment, a plurality of doses of the medicament are administered. The frequency of administration of the medicament may vary according to any one of a variety of factors, such as the severity of the symptoms, etc. For example, in an embodiment, the medicament may 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), every day (qd), twice a day (qid), or three times a day (tid). As discussed above, in an embodiment, the medicament is administered continuously.

[0259] The duration of administration of an agent, e.g., the period of time over which the agent is administered, can vary depending on any of a variety of factors, e.g., patient response, etc. For example, the combination or individual agents can be administered for a period of about one day to one week, about two weeks to four weeks, about one month to two months, about two months to four months, about four months to six months, about six months to eight months, about eight months to 1 year, about 1 year to 2 years, or about 2 years to 4 years, or longer.

[0260] Doses up to 60 μg / kg are non-toxic. Lower concentrations, such as 1-4 μg / kg, have also shown biological activity in in vivo systems. -9 -10 -6 M is effective and is expected to be achieved in the cellular environment. (See 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 TK.,Janjetovic Z,Tuckey RC,Bieniek,R,Yue Y,Li W,Chen J,Miller D,Chen T,Holick M(2011)20-hydroxyvitaminD2 is a non-calcemic analog of vitamin D with potent antiproliferative andprodifferentiation activities in normal and malignant cells.Am JPhysiol:CellPhysiol 300:C526-C541; Wang J,Slominski AT,Tuckey RC,Janjetovic Z,Kulkarni A,Chen J,Postlethwaite A,Miller D,Li W(2012)20-Hydroxylvitamin D3 possesseshigh efficacy against proliferation of cancer cells while being non-toxic.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.JClin 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 asPotential Therapeutics:The Metabolism,Toxicity Profiling,andAntiproliferative Activity.Anticancer Res 34:2153-2163. ).

[0261] In one aspect, the dosage of any single agent disclosed herein for administration to a subject (e.g., a mammal, such as a human) having a condition (e.g., COVID-19) is about 2 to 60 micrograms / kg, or a combination of agents: each agent can be about 2 to 60 micrograms / kg.

[0262] Route of administration

[0263] Embodiments of the present disclosure provide methods and compositions for administering pharmaceutical agents (e.g., modified saponins) to a subject (e.g., a human) using any available methods and routes suitable for drug delivery, including in vivo and ex vivo methods, as well as systemic and local administration routes.

[0264] Administration routes include intranasal, intramuscular, intratracheal, subcutaneous, intradermal, topical, intravenous, rectal, nasal, oral and other enteral and parenteral administration routes. If desired, administration routes can be combined or adjusted according to the antibody and / or desired effect. The medicament can be administered in single dose or multiple dose form.

[0265] Embodiments of the pharmaceutical agents may be administered to an individual using available conventional methods and routes suitable for delivery of conventional drugs, including systemic or topical routes. In general, routes of administration contemplated by the present disclosure include, but are not limited to, enteral, parenteral, or inhalation routes.

[0266] Parenteral routes of administration other than inhalation administration include, but are not limited to, topical, transdermal, subcutaneous, intramuscular, intraorbital, intracapsular, intraspinal, intrasternal, and intravenous routes, i.e., any route of administration other than through the digestive tract. Parenteral administration can be performed to achieve systemic or local delivery of the medicament. When systemic delivery is desired, administration generally involves invasive or systemically absorbed local or mucosal administration of the drug preparation.

[0267] In embodiments, the agent may also be delivered to the subject by enteral administration. Enteral administration routes include, but are not limited to, oral and rectal (eg, using a suppository) delivery.

[0268] Methods for administering agents through the skin or mucous membranes include, but are not limited to, topical application of suitable pharmaceutical preparations, transdermal delivery, injection, and epidermal administration. For transdermal delivery, absorption enhancers or iontophoresis are suitable methods. Iontophoresis delivery can be achieved using commercially available "patches" that deliver their products via electrical pulses through unbroken skin for several days or longer.

[0269] Although the embodiments of the present disclosure are described in conjunction with examples and corresponding text and drawings, it is not intended to limit the present disclosure to the embodiments described in these descriptions. On the contrary, it is intended to cover all alternatives, modifications and equivalents included in the spirit and scope of the embodiments of the present disclosure.

[0270] Although the embodiments of the present disclosure are described in conjunction with examples and corresponding text and drawings, it is not intended to limit the present disclosure to the embodiments described in these descriptions. On the contrary, it is intended to cover all alternatives, modifications and equivalents included in the spirit and scope of the embodiments of the present disclosure.

[0271] Example 1

[0272]

[0273] Reagents and conditions: (a) Boc2O, NaHCO3, DMF, 0℃-room temperature; (b) BzCl, Et3N, DCM; (c) BnBr, NaH, DMF, 0℃-room temperature;

[0274] (d) TFA, DCM; (e) NMM, HOBt, EDC.HCl, R'NH2, H2O / EtOH, room temperature; (f) Pd / C, H2 (55 psi), MeOH / H2O;

[0275] (g) Benzoyl NHS ester, methanol.

[0276] 2 synthesis.

[0277] At 0°C, (BOC)2O (33 mg, 0.15 mmol) was added to 11-amino-1-undecanol (20 mg, 0.1 mmol) stirred in N,N-dimethylformamide (0.5 mL) and NaHCO3 (5 mg, 20%, v / v). The reaction mixture was stirred at room temperature overnight. Afterwards, the reaction mixture was filtered by column chromatography filled with silica gel and ethyl acetate to give 2, which was used directly in the synthesis of 3a and 3b.

[0278] Synthesis of 3a.

[0279] Benzoyl chloride (230 μl, 2 mmol) was added to intermediate 2 (30 mg, 0.1 mmol) stirred in a mixture of dichloromethane (2.25 mL) and triethylamine (1.39 mL, 10 mmol) at room temperature. The mixture was stirred for 48 hr and then concentrated using a rotary evaporator to extract with EtOAc. The organic layer was washed three times with NaHCO3 and dried over Na2SO4. The collected organic layer was then purified by silica gel column chromatography (eluted with petroleum ether and ethyl acetate gradient) to give 3b (15 mg, 38%). Rf (PE / EtOAc=9:1)=0.49.1 H 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 C 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: for C 18 H 30 [M+H-Boc] of NO2 + Calculated value, 292.2277; measured value, 292.2269.

[0280] Synthesis of 3b.

[0281] 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 an ice bath in dry N,N-dimethylformamide (1.5 mL) for 15 min. The mixture was stirred at room temperature overnight. The mixture was then extracted with EtOAc and washed three times with deionized water. The collected organic layer was dried over sodium sulfate. Afterwards, the organic layer was concentrated by rotary evaporator, and the mixture was then purified by silica gel column chromatography (eluted with petroleum ether and ethyl acetate gradient) to produce the desired compound 3b (29 mg, 50%). f (PE / EtOAc=9:1)=0.71 1 H NMR (700MHz, CD3OD) (characteristic proton): δ7.35-7.32 (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); 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: for C 18 H 32 [M+H-Boc] of NO + Calculated value, 278.2484; measured value, 278.2480.

[0282] Synthesis of 4a.

[0283] Trifluoroacetic acid (85 μl, 30%, v / v) was added to a mixture of 3a (20 mg, 0.07 mmol) and anhydrous dichloromethane (1 mL) to deprotect the boc protecting group. The mixture was stirred for 3 hr and quenched with Na2CO3. The mixture was concentrated by rotary evaporator to give the product (6.5 mg, 33%)

[0284] Synthesis of 4b.

[0285] 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 by rotary evaporator to give product 4b (12 mg, 82%).

[0286] General procedure for derivatization MS II (not optimized):

[0287] To MS II in ethanol / water was added side chains, N-methylmorpholine (NMM), hydroxybenzotriazole (HOBt), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl) at room temperature. The reaction mixture was stirred for 3 days and then purified by using a semi-preparative C 18 , 250x10 mm, 5 micron column and H2O / MeCN gradient (90%-10% H2O in 45 minutes, flow rate 3 mL / min) were directly purified by RP HPLC. The product fractions were concentrated on a rotary evaporator at room temperature to remove MeCN, and then the remaining water was removed on a freeze dryer to provide the derivative as a white solid.

[0288] For 1D (12.4 mg, 35%): 1H NMR (500 MHz, CD3OD) (characteristic protons): δ9.50 (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. .9Hz,1H),4.56(d,J=7.8Hz,1H),4.52(b,1H),4.50-4.40(m,3H),4.35(t,J=6.6Hz,2H),4.24(dd,J=2.8,1.9Hz,1H),4. 06-4.01(m,2H),2.30(t,J=13.6Hz,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); 13 CNMR (176.0MHz, CD3OD): δ209.6,175.5,169.8,166.7,143.6,132.8,130.2,129.1,128.2,121.5,104.6,103.7,103.5,102.8, 101.9,99.3,94.0,87.4,84.6,84.5,82.2,77.3,76.8,76.5,76.0,75.4,75.1,74.2,74.0,73.6,73.3,73.0,72.4,72.3,71.5,7 0.8,70.6,70.5,70.0,68.0,67.4,65.7,64.9,60.8,60.6,54.8,41.5,41.0,39.7,38.7,38.0,35.7,35.2,32.0,30.5,29.9,29.5,29.3,29.2,29.0,28.9,28.4,26.6,26.0,25.8,24.6,23.3,23.0,20.0,17.0,16.5,16.4,15.1,9.6; HRMS (ESI-TOF) m / z: for C 94 H 148 NO 42 [M+H] + Calculated value, 1962.9476; Found, 1962.9467. For 1E (8.6 mg, 56% from 1H and benzoyl N-hydroxysuccinimide ester): 1HNMR (600 MHz, CD3OD) (characteristic protons): δ9.49 (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, 1 H),4.50-4.46(m,2H),4.45(d,J=7.7Hz,1H),4.25(dd,J=2.9,2.2Hz,1H),4.06-4.01(m,2H),3.18(t,J=11.3Hz,1H),3.14(dd,J=9.4 ,8.1Hz,1H),2.90(dd,J=14.6,4.5Hz,1H),2.30(t,J=13.7Hz,1H),1.18(s,3H),1.03(s,3H),0.93(s,3H),0.88(s,3H),0.81(s,3H); 13 CNMR (150.9MHz, CD3OD): δ209.6,175.6,169.8,168.8,143.5,134.5,131.1,128.2,126.8,121.6,104.7,103.8,103.5,102.8,102.7,10 1.9,99.3,94.0,87.4,84.6,84.5,82.2,77.3,76.8,76.4,76.0,75.4,75.1,74.2,74.0,73.6,73.4,73.0,72.4,72.3,71.6,71.5,70.8, 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: for C 93 H 147 N2O 41 [M+H] +Calculated value, 1947.9479; measured value, 1947.9386.

[0289] For 1F (8.3 mg, 36%): 1 HNMR (600MHz, CD3OD) (characteristic proton): δ9.50 (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. 1Hz,1H),4.56(d,J=8.0Hz,1H),4.25(dd,J=2.9,1.7Hz,1H),2.91(dd,J=14.2,4.5Hz,1H) ,2.30(t,J=13.6Hz,1H),1.19(s,3H),1.03(s,3H),0.94(s,3H),0.88(s,3H),0.81(s,3H); 13 C NMR (176.0MHz, CD3OD): δ209.6,175.5,169.7,143.6,138.5,121.5,104.7,103.8,103.5,102.8,102.7,101.9,99.3,94 .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 (ESITOF) m / z: for C 94 H 150 NO 41 [M+H] + Calculated value, 1948.9383; measured value, 1948.9684.

[0290] For 1G (7.4 mg, 67% from 1F): 1HNMR (600MHz, CD3OD) (characteristic proton): δ9.49 (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=2.8,1.8Hz,1H),3.17(t,J=10.9Hz,1H),3.12(dd,J=8.2,8.1Hz,1H),2.90(dd,J=14.0,4.4Hz, 1H), 2.30 (t, J = 13.6Hz, 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 C NMR (213.8MHz, CD3OD): δ209.6,175.6,169.8,143.5,121.5,104.7,103.8,103.6,102.8,102.7,101.9,99.3,94.0,87.4,84.6,84.5, 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: for C 87 H 144 NO 41 [M+H]+ calculated, 1858.9214; found, 1858.9211.

[0291] For 1H (20.4 mg, 27%): 1H NMR (600MHz, CD3OD) (characteristic proton): δ9.50 (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=7.7Hz,1H),4.25(dd,J=3.1,1.9Hz,1H),4.05-4.01(m,2H),3.18(t,J=11.1Hz,1H),3.14(dd,J= 9.4,8.1Hz,1H),2.31(t,J=13.6Hz,1H),1.20(s,3H),1.03(s,3H),0.96(s,3H),0.89(s,3H),0.81(s,3H); 13 C NMR (150.9MHz, CD3OD): δ209.6,175.6,169.8,143.6,121.5,104.7,103.8,103.5,102.8,101.7,101.9,99.3,94.0,87.4,84.6,82 .1,77.3,76.9,76.4,76.0,75.4,75.1,74.3,74.0,73.6,73.3,73.0,72.4,72.3,71.7,71.5,70.8,70.6,70.5,70.1,70.0,69.6,6 9.2,69.1,68.0,67.4,65.7,60.8,60.7,54.8,48.5,46.6,41.5,41.0,39.7,39.4,38.7,37.9,35.7,35.2,32.7,32.0,30.5,29.9,29.2,29.0,28.9,28.8,27.2,26.4,26.0,25.9,24.5,23.2,23.1,20.0,17.0,16.5,16.4,15.1,15.0,9.6; HRMS (ESI-TOF) m / z: for C 86 H 143 N2O 40 [M+H] + Calculated value, 1843.9217; measured value, 1843.9216.

[0292] Example 2

[0293]

[0294] General procedure for preparation of side chains 5D to 5H (not optimized):

[0295] First, a mixture of 11-aminoundecanoic acid (50 mg, 0.25 mmol) and thionyl chloride (36 μL, 0.5 mmol) was stirred at room temperature, and then heated to 70 ° C in an oil bath and stirred for about 1 hr. 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 DCM / MeOH gradient) to produce the desired side chain.

[0296] 5D synthesis.

[0297] 3,5-Dimethoxybenzyl alcohol (84.0 mg, 0.5 mmol) was used and purified by silica column chromatography (eluting with a DCM / MeOH gradient) to give 5D (41.0 mg, 47%). f (DCM / MeOH 9:1) 0.43; 1 H 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 C 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: for C 20 H 34 [M+H] of NO4 + Calculated value, 352.2488; measured value, 352.2486.

[0298] Synthesis of 5E.

[0299] 3-Methoxybenzyl alcohol (63.0 mg, 0.5 mmol) was used and purified by silica gel column chromatography (eluting with a DCM / MeOH gradient) to give 5E (65.0 mg, 81%). f(DCM / MeOH 9:1) 0.56; 1H NMR (700MHz, CD3OD): δ7.26 (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 C 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: for C 19 H 32 [M+H] of NO3 + Calculated value, 322.2382; measured value, 322.2380.

[0300] Synthesis of 5F:

[0301] 3-Phenoxyphenylmethanol (87.0 μL, 0.5 mmol) was used and purified by silica gel column chromatography (eluting with a DCM / MeOH gradient) to give 5F (50.0 mg, 52%). f (DCM / MeOH 9:1) 0.43; 1 H NMR (700 MHz, CD3OD) δ7.32 (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); 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: for C 24 H 34 [M+H] of NO3 + Calculated value, 384.2539; measured value, 384.2535.

[0302] 5G Synthesis:

[0303] Purification using silica gel column chromatography (eluting with a DCM / MeOH gradient) using 3-nitrobenzyl alcohol (60.0 mg, 0.5 mmol) gave 5G (64.0 mg, 76%). f (DCM / MeOH 9:1) 0.40; 1 H NMR (700 MHz, CD3OD): δ8.21 (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); 13 C 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: for C 18 H 29 [M+H] of N2O4 + Calculated value, 337.2127; measured value, 337.2126.

[0304] Synthesis of 5H:

[0305] 5H (49.0 mg, 64%) was obtained using 4-fluorobenzyl alcohol (55.0 μL, 0.5 mmol) and purified by silica gel column chromatography (eluting with a DCM / MeOH gradient). f (DCM / MeOH 9:1) 0.37; 1 H NMR (700 MHz, CD3OD): δ7.31 (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 C 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: for C 18 H 29 [M+H] of FNO2 + Calculated value, 310.2182; measured value, 310.2179.

[0306] General procedure for derivatization MS II (not optimized):

[0307] At room temperature, add side chain (5D to 5H) (0.018mmol), N-methylmorpholine (NMM) (10.0μl, 0.09mmol), hydroxybenzotriazole (HOBt) (8.3mg, 0.054mmol) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC HCl) (11.0mg, 0.054mmol) in MS II (15mg, 0.009mmol) in ethanol / water (1:2, 0.5mL). The reaction mixture is stirred for 3 days, and then directly purified with RP HPLC by using semi-preparative C18, 250x10mm, 5 micron column and H2O / MeCN gradient (90%-10% H2O in 45 minutes, flow rate 3mL / min). The product fraction is concentrated on a rotary evaporator at room temperature to remove MeCN, and then the remaining water is removed on a freeze dryer to provide a derivative in the form of a white solid.

[0308] For 2D, 5D (6.3 mg, 0.018 mmol) was used to provide the final product 2D (10 mg, 56%); 1 H NMR (600 MHz, CD3OD) (characteristic proton): δ9.47 (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=7.6Hz,1H),4.23(dd,J=3.0,1.8Hz,1H),3.15(t,J=11.0Hz,1H),3.13(dd,J=9.4,8.1Hz,1H),2.88(dd,J= 14.1,4.5Hz,1H),2.38(t,J=7.3Hz,2H),2.27(t,J=13.4Hz,1H),1.16(s,3H),0.99(s,3H),0.92(s,3H),0.86(s,3H),0.77(s,3H); 13 CNMR (176.0MHz, CD3OD): δ209.6,175.5,173.8,169.8,161.0,143.5,138.6,121.5,105.5,104.7,103.8,103.5,102.8,101.9,99.5,99 .3,94.0,87.4,84.6,84.5,82.2,77.3,76.8,76.4,76.0,75.4,75.1,74.2,74.0,73.6,73.4,73.0,72.4,72.3,71.5,70.8,70.7,70.5,7 0.1,70.0,69.6,69.2,69.1,68.0,67.4,65.7,65.6,60.8,60.6,54.8,54.4,41.5,41.0,39.7,38.7,38.0,35.7,35.3,33.7,32.8,32.0,30.5,29.9,29.4,29.2,29.1,29.0,28.9,28.7,26.5,25.9,24.7,23.3,23.1,20.0,17.0,16.5,16.4,15.1,9.6; HRMS (ESI-TOF) m / z: for C 96 H 152 NO44 [M+H] + Calculated value, 2022.9687; measured value, 2022.9675.

[0309] For 2E, 5E (6 mg, 0.018 mmol) was used to provide the final product 2E (8 mg, 45%); 1 H NMR (600MHz, CD3OD): δ9.47(s,1H),7.26(t,J=7.9Hz,1H),6.93-6.89(m,2H),6.87(dd,J=8.2,2.7Hz,1H),5.40(d,J=1.8Hz,1H),5.30(t,J=3.7 Hz,1H),5.20(d,J=8.2Hz,1H),5.09(s,2H),5.01(d,J=1.6Hz,1H),4.71(d,J=7.9Hz,1H),4.53(d,J=7.9Hz,1H),4.49(b,1H),4.42(d,J=7.7Hz, 1H),4.22(dd,J=3.1,1.9Hz,1H),3.15(t,J=10.8Hz,1H),3.11(dd,J=9.3,8.0Hz,1H),2.88(dd,J=14.3,4.1Hz,1H),2.37(t,J=7.3Hz,2H),2.27 (t,J=13.6Hz,1H),1.39(s,3H),1.22(d,J=6.3Hz,3H),1.20(d,J=6.5Hz ,3H),1.16(s,3H),1.00(s,3H),0.91(s,3H),0.85(s,3H),0.78(s,3H); 13C NMR (176.0MHz, CD3OD): δ209.6,175.5,173.8,169.8,161.4,160.0,143.5,137.8,129.2,121.5,119.9,113.2,104.7,103.8,103. 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: for C 95 H 149 NO 43 [M+H]+ calculated, 1992.9582; found, 1992.9662.

[0310] For 2F, 5F (7 mg, 0.018 mmol) was used to provide the final product 2F (10.0 mg, 54%): 1H NMR(600MHz,CD3OD):δ9.51(s,1H),7.42-7.30(m,3H),7.17(tt,J=7.4,2.1Hz,1H),7.14(d,J=3.7Hz,1H),7.05-7.01(m,3H),6.97(dd,J=8.1,2.3Hz,1H),5.45(d,J=1.6Hz,1H),5.34(t,J=3.5Hz,1H),5.25(d,J=8.3Hz,1H),5.14(s,2H),5.06(d,J=1.5Hz,1H),4.76(d,J=7.9Hz,1H),4.58(d,J=7.9Hz,1H),4.55(b,1H),4.47(d,J=7.7Hz,1H),4.27(dd,J=3.0,1.9Hz,1H),4.07-4.04(m,2H),3.20(t,J=11.0Hz,1H),3.16(dd,J=9.4,8.0Hz,1H),2.93(dd,J=14.1,4.4Hz,1H),2.40(t,J=7.3Hz,2H),2.32(t,J=13.6Hz,1H),1.44(s,3H),1.27(d,J=6.2Hz,3H),1.25(d,J=6.4Hz,3H),1.21(s,3H),1.04(s,3H),0.96(s,3H),0.90(s,3H),0.82(s,3H); 13C NMR (150.9MHz, CD3OD): δ209.6,175.5,173.7,169.8,157.6,157.1,138.6,129.7,129.6,123.2,122.4,121.5,118.7,117.9,117.7,104.7,103 .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: for C 100 H 152 NO 43 [M+H] + Calculated value, 2054.9738; Found, 2054.9714. For 2G, 5G (6 mg, 0.018 mmol) was used to provide the final product 2G (10.0 mg, 56%): 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: For C 94 H 147 N2O44 [M+H] + Calculated value, 2007.9327; measured value, 2007.9337.

[0311] For 2H, 5H (5.6 mg, 0.018 mmol) was used to provide the final product 2H (10.0 mg, 56%): 1 H NMR (600MHz, CD3OD): δ9.51(s,1H),7.45-7.40(m,2H),7.11(tt,J=8.7,2.1Hz,1H),5.44(d,J=1.6Hz,1H),5.34(t,J=3.4Hz,1H),5.24(d, J=8.3Hz,1H),5.13(s,2H),5.05(d,J=1.6Hz,1H),4.75(d,J=8.0Hz,1H),4.57(d,J=7.8Hz,1H),4.54(b,1H),4.46(d,J=7.6Hz,1H),4.26( dd,J=3.0,2.1Hz,1H),3.19(t,J=11.0Hz,1H),3.15(dd,J=9.2,8.3Hz,1H),2.92(dd,J=14.5,4.4Hz,1H),2.39(t,J=7.5Hz,2H),2.31(t,J =13.6Hz,1H),1.44(s,3H),1.26(d,J=6.3Hz,3H),1.25(d,J=6.4Hz,3H),1.21(s,3H),1.04(s,3H),0.95(s,3H),0.89(s,3H),0.82(s,3H); 13C NMR (176.0MHz, CD3OD): δ209.6,175.5,173.7,169.8,163.3,161.9,143.6,132.5,130.2,130.1,114.9,114.8,104.7,103.7,103.5,102.8 ,101.9,99.3,94.0,87.4,84.6,84.5,82.2,77.3,76.8,76.4,76.0,7 5.4,75.1,74.2,74.0,73.6,73.3,73.0,72.4,72.3,71.5,70.8,70.7, 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: for C 94 H 147 FNO 42 [M+H] + Calculated value, 1980.9382; measured value, 1980.9463.

[0312] antigen:

[0313] Chicken egg albumin (Vac-pova) for in vivo use was purchased from InvivoGen.

[0314] Mice and immunization:

[0315] The BALB / c mice used in this study were from Frederick Cancer Research (Fredrick, MD). In order to evaluate the adjuvant activity of the immunoadjuvant based on MS saponin, female mice groups (8-10 weeks old; 6 mice per group) were immunized by subcutaneous (sc) route using OVA (20 μg) alone or using antigen plus appropriate adjuvant (such as QS-21 (20 μg) or MS adjuvant (50-100 μg)) on day 0, day 14 and day 28. Before each immunization and two weeks after the last immunization, mice were weighed and blood samples were collected from the lateral tail vein using a heparinized capillary pipette. Serum was obtained after centrifugation and stored at -20°C until determination.

[0316] Assessment of Antibody Response:

[0317] The levels of anti-OVA-specific serum IgG and IgG subclasses in each group were determined by enzyme-linked immunosorbent assay (ELISA). Maxisorp microtiter plates (NUNC International, Roskilde, DK) were incubated with rHagB (1 μg / ml), OVA (0.1 μg / ml), or borate-buffered saline (BBS; 100 mM NaCl, 50 mM boric acid, 1.2 mM Na2B4O 7, , pH 8.2) in the optimal amount of goat anti-mouse IgG, IgG1 or IgG2a at 4 ° C overnight coating. The plate is blocked at room temperature for 2h with 1% bovine serum albumin (BSA) and 0.02% sodium azide in BBS. Serum samples diluted twice continuously are added to the plate in duplicate. In order to generate a standard curve, serially diluted mouse immunoglobulin reference serum (MP Biomedicals, Solon, OH) is added to two rows of holes on each plate, which have been coated with appropriate anti-mouse IgG or IgG subclass reagents. After incubation (4 ° C overnight) and washing the plate, goat anti-mouse IgG or IgG subclass antibodies conjugated with horseradish peroxidase are added to appropriate holes. After incubation at room temperature for 4h, the plate is washed and developed with o-phenylenediamine substrate and hydrogen peroxide. Color development is recorded at 490nm. Antibody concentrations were determined by interpolation of 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).

[0318] Figures 5A to 5C Serum IgG, IgG1 and IgG2a anti-OVA responses in mice immunized by subcutaneous route with ovalbumin (OVA) alone, or with QS-21 or VSA-2, or with one of the saponin adjuvants 1D to 1H are shown. Statistical significance was assessed by t-test (using unpaired, nonparametric and Mann-Whiteny test). *P<0.05, **P<0.01.

[0319] Figures 6A to 6C Serum IgG, IgG1 and IgG2a anti-OVA responses in mice immunized by subcutaneous route with ovalbumin (OVA) alone, or with QS-21 or VSA-2, or with one of the saponin adjuvants 2D to 2H are shown. Statistical Analysis:

[0320] The statistical significance of antibody responses was assessed by t-test (using unpaired, nonparametric and Mann-Whiteny tests) using GraphPad Prism 8. Differences were considered significant when P values ​​< 0.05.

[0321] in conclusion:

[0322] Many MS saponin-derived immunostimulants have been prepared and evaluated. These MS derivatives are prepared by incorporating terminal functionalized side chains into the C3 glucuronic acid unit of the natural saponin MS II via an amide formation reaction. These non-natural semisynthetic saponins show potent immunostimulatory activity comparable to that of the FDA-approved saponin immunostimulant QS-21.

[0323] References for Example 1

[0324] 1.Brunner R, Jensen-Jarolim E, Pali-Scholl I.The ABC of clinical and experimental adjuvants-a brief overview.Immunol Lett.2010;128(1):29-35.doi:10.1016 / j.imlet.2009.10.005.PubMed PMID:19895847; PMCID:2999744.

[0325] 2. Kensil CR, Mo AX, Truneh A. Current vaccine adjuvants: An overview of adiverse class. Frontiers in Bioscience. 2004; 9: 2972-88.

[0326] 3. Leroux-Roels G. Unmet needs in modern vaccinology: adjuvants to improve the immune response. Vaccine. 2010; 28 Suppl 3: C25-36. doi: 10.1016 / j.vaccine.2010.07.021. PubMed PMID: 20713254.

[0327] 4. Sharp FA, Lavelle EC. Discovery of vaccine adjuvants. In: Gad SC, ed. Development of Therapeutic Agents Handbook. 1st ed. Hoboken, NJ: John Wiley & Sons, Inc; 2012. pp. 533-46.

[0328] 5. Wang W, Singh M. Selection of adjuvants for enhanced vaccine potency. World J Vaccines. 2011;01(02):33-78. doi:10.4236 / wjv.2011.12007.

[0329] 6. Weeratna RD, McCluskie MJ. Recent advances in vaccine adjuvants. Source: Miller AA, Miller PF, editors. Emerging Trends in Antibacterial Discovery: Answering the Call to Arms. Great Britain: Caister Academic Press; 2011. pp. 303-22.

[0330] 7. Coffman RL, Sher A, Seder RA. Vaccine adjuvants: putting innate immunity to work. Immunity. 2010;33(4):492-503. doi:10.1016 / j.immuni.2010.10.002. PubMed PMID: 21029960; PMCID: 3420356.

[0331] 8. Banday AH, Jeelani S, Hruby VJ. Cancer vaccine adjuvants--recent clinical progress and future perspectives. Immunopharmacol Immunotoxicol. 2015;37(1):1-11. Epub 2014 / 10 / 17. doi:10.3109 / 08923973.2014.971963. PubMed PMID: 25318595.

[0332] 9.Bastola R,Noh G,Keum T,Bashyal S,Seo JE,Choi J,Oh Y,Cho Y,LeeS.Vaccine adjuvants:smart components to boost the immune system.Archives ofpharmacal research.2017;40(11):1238-48.Epub 2017 / 10 / 14.doi:10.1007 / s12272-017-0969-z.PubMed PMID:29027637。

[0333] 10.Di Pasquale A,Preiss S,Tavares Da Silva F,Garcon N.VaccineAdjuvants:from 1920 to 2015 and Beyond.Vaccines(Basel).2015;3(2):320-43.Epub2015 / 09 / 08.doi:10.3390 / vaccines3020320.PubMed PMID:26343190;PMCID:PMC4494348。

[0334] 11.Temizoz B,Kuroda E,Ishii KJ.Vaccine adjuvants as potential cancerimmunotherapeutics.Int Immunol.2016;28(7):329-38.Epub 2016 / 03 / 24.doi:10.1093 / intimm / dxw015.PubMed PMID:27006304;PMCID:PMC4922024。

[0335] 12.Bonam SR,Partidos CD,Halmuthur SKM,Muller S.An overview of noveladjuvants designed for improving vaccine efficacy.Trends in PharmacologicalSciences.2017;38(9):771-93。

[0336] 13. O’Hagan DT, Friedland LR, Hanon E, Didierlaurent AM. Towards an evidence based approach for the development of adjuvanted vaccines. Current Opinion in Immunology. 2017;47:93–102.

[0337] 14. N, Leroux-Roels G, Cheng W-F. Vaccine Adjuvants. Source: N, Stern PL, Cunningham AL, eds. Understanding Modern Vaccines Perspectives in Vaccinology Amsterdam: Elsevier; 2011. pp. 89-113.

[0338] 15. Giudice GD, Rappuoli R, Didierlaurent AM. Correlates of adjuvanticity: A review on adjuvants in licensed vaccines. Semin Immunol. 2018;2018:14-21.

[0339] 16. Shi S, Zhu H, Xia X, Liang Z, Ma X, Sun B. Vaccine adjuvants: Understanding the structure and mechanism of adjuvanticity. Vaccine. 2019;37:3167-78.

[0340] 17.Rivera-Hernandez T,Pandey M,Henningham A,Cole J,Choudhury B,CorkAJ,GillenCM,Ghaffar KA,West NP,Silvestri G,Good MF,Moyle PM,Toth I,Nizet V,Batzloff MR,Walker MJ.Differing Efficacies of Lead Group A StreptococcalVaccine Candidates and Full-Length M Protein in Cutaneous and InvasiveDisease Models.mBio.2016;7(3):e00618-16.doi:10.1128 / mBio.00618-16。

[0341] 18.Rivera-Hernandez T,Rhyme MS,Cork AJ,Jones S,Segui-Perez C,BrunnerL,Richter J,Petrovsky N,Lawrenz M,Goldblatt D,Collin N,Walker MJ.Vaccine-Induced Th1-TypeResponse Protects against Invasive Group A StreptococcusInfection in the Absence ofOpsonizing Antibodies.mBio.2020;11(2).Epub 2020 / 03 / 12.doi:10.1128 / mBio.00122-20.PubMed PMID:32156809;PMCID:PMC7064752。

[0342] 19.Didierlaurent AM,Laupeze B,Di Pasquale A,Hergli N,Collignon C,Garcon N.Adjuvantsystem AS01:helping to overcome the challenges of modernvaccines.Expert Rev Vaccines.2017;16(1):55-63.Epub 2016 / 07 / 28.doi:10.1080 / 14760584.2016.1213632.PubMed PMID:27448771。

[0343] 20.James SF,Chahine EB,Sucher AJ,Hanna C.Shingrix:the new adjuvantedrecombinant herpes zoster vaccine.Annals of Pharmacotherapy.2018;52(7):673-80。

[0344] 21.Cunningham AL,Lal H,Kovac M,Chlibek R,Hwang SJ,Díez-Domingo J,Godeaux O,Levin MJ,McElhaney JE,Puig-BarberaJ,VandenAbeele C,Vesikari T,Watanabe D,Zahaf T,Ahonen-Ahanen-Gomezarba,B. JF,Campora L,deLooze F,DowneyHJ,Ghesquiere W,Gorfinkel I,Korhonen T,Leung E,McNeil SA,Oostvogels L,RomboL,Smetana J,Weckx L,Yeo W,Heineman TC. New England Journal ofMedicine.2016;375(11):1019-32。

[0345] 22.Lacaille-Dubois MA.Updated insights into the mechanism of actionand clinical profileof the immunoadjuvant QS-21:A review.Phytomedicine.2019;60:152905.Epub 2019 / 06 / 12.doi:10.1016 / j.phymed.2019.152905.PubMed PMID:31182297;PMCID:PMC7127804。

[0346] 23.Martin RS,Briones R.Industrial uses and sustainable supply ofQuillaja Saponaria(Rosaceae)Saponins.Economic Botany.1999;53(3):302-11。

[0347] 24.Ragupathi G,Gardner JR,Livingston PO,Gin DY.Natural and syntheticsaponin adjuvantQS-21 for vaccines against cancer Expert Rev Vaccines.2011;10:463-70。

[0348] 25.Wang P,Dai Q,Thogaripally P,Zhang P,Michalek SM.Synthesis of QS-21-basedimmunoadjuvants.J Org Chem.2013;78:11525-34。

[0349] 26.Wang P,Devalankar DA,Dai Q,Zhang P,Michalek SM.Synthesis andEvaluation of QS-21-Based Immunoadjuvants with a Terminal-Functionalized SideChain Incorporated in the West Wing Trisaccharide.J Org Chem.2016;81:9560-6。

[0350] 27.Wang P,Ding X,Kim H, Michalek SM,Zhang P.Vaccineadjuvants derivatized from Momordica saponins I and II.J Med Chem.2019;62:9976-82。

[0351] 28.Wang P, Sui X,Zhang P,Michalek SM.Synthesis andevaluation of a QS-17 / 18-based vaccine adjuvant.J Med Chem.2019;62:1669-76.doi:10.1021 / acs.jmedchem.8b01997。

[0352] 29.Wang P, Sui X,Zhang P,Michalek SM.Synthesis andevaluation of QS-7-based vaccine adjuvants.ACS Infectious Diseases.2019;5:974-81.doi:10.1021 / acsinfecdis.9b00039。

[0353] 30.Wang P.Natural and Synthetic Saponins as VaccineAdjuvants.Vaccines(Basel).2021;9(3):222.doi: https: / / doi.org / 10.3390 / vaccines9030222 。

[0354] 31.Wang P,Ding X,Kim H,Michalek SM,Zhang P.Structural Effect onAdjuvanticity of Saponins.J Med Chem.2020;63:3290-7。

Claims

1. A modified saponin having the formula: in: 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 fucosyl unit, wherein f3 and f4 may form a cyclic ketal ring or a cyclic carbonate; f5 is selected from H, a methyl group, a -CH2OH group, R4-NR5-C(O)-, R4-OC(O)- or R4-O-CH2-; wherein R4 and R5 are each independently a compound having the structure R6[(CX2) 0-20 O 0-1 (CY2) 0-20 ] 0-20 A linear chain of wherein X and Y are each independently H or a halogen atom; wherein R6 is COOR7, C(O)NR7R8, NR7R8 or OR7; Where R7 is Ar[(CZ2) 0-20 O 0-1 (CL2) 0-20 ] 0-20 , wherein R8 is H or alkyl; wherein Z and L are each independently H or a halogen atom; wherein Ar is a substituted or unsubstituted aromatic system; ga5 is selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-; Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 A linear chain of wherein X1 and Y1 are each independently H or a halogen atom; Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ; Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl; Wherein Z1 and L1 are each independently H or a halogen atom; wherein Ar2 is a substituted or unsubstituted aromatic group; r3 is H, a monosaccharide, a disaccharide or a trisaccharide; x3 is H, a monosaccharide (except xylose) or a disaccharide; and ga3 is H, monosaccharide or disaccharide.

2. A modified saponin having formula I: in: 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 selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-; Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 A linear chain of wherein X1 and Y1 are each independently H or a halogen atom; Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ; Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl; wherein Z1 and L1 are each independently H or a halogen atom; and Wherein Ar2 is a substituted or unsubstituted aromatic group.

3. A modified saponin having one of formula 2D to 2H:

4. A pharmaceutical composition comprising a therapeutically effective amount of a modified saponin having the formula: in: 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 fucosyl unit, wherein f3 and f4 may form a cyclic ketal ring or a cyclic carbonate; f5 is selected from H, a methyl group, a -CH2OH group, R4-NR5-C(O)-, R4-OC(O)- or R4-O-CH2-; wherein R4 and R5 are each independently a compound having the structure R6[(CX2) 0-20 O 0-1 (CY2) 0-20 ] 0-20 A linear chain of wherein X and Y are each independently H or a halogen atom; wherein R6 is COOR7, C(O)NR7R8, NR7R8 or OR7; Where R7 is Ar[(CZ2) 0-20 O 0-1 (CL2) 0-20 ] 0-20 , wherein R8 is H or alkyl; wherein Z and L are each independently H or a halogen atom; wherein Ar is a substituted or unsubstituted aromatic system; ga5 is selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-; Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 A linear chain of wherein X1 and Y1 are each independently H or a halogen atom; Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ; Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl; Wherein Z1 and L1 are each independently H or a halogen atom; wherein Ar2 is a substituted or unsubstituted aromatic group; r3 is H, a monosaccharide, a disaccharide or a trisaccharide; x3 is H, a monosaccharide (except xylose) or a disaccharide; and ga3 is H, monosaccharide or disaccharide.

5. A pharmaceutical composition comprising a therapeutically effective amount of a modified saponin having formula I for treating a condition: in: 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 selected from R 14 -NR 15 -C(O)-、R 14 -OC(O)- or R 14 -O-CH2-; Where R 14 and R 15 Each independently has the structure R 16 [(CX12) 0-20 O 0-1 (CY12) 0-20 ] 0-20 A linear chain of wherein X1 and Y1 are each independently H or a halogen atom; Where R 16 For COOR 17 、C(O)NR 17 R 18 NR 17 R 18 OR 17 ; Where R 17 Ar2[(CZ12) 0-20 O 0-1 (CL12) 0-20 ] 0-20 , where R 18 is H or alkyl; wherein Z1 and L1 are each independently H or a halogen atom; and Wherein Ar2 is a substituted or unsubstituted aromatic group.

6. A pharmaceutical composition comprising a therapeutically effective amount of a modified saponin having one of Formulae 2D to 2H to treat a disorder:

7. The pharmaceutical composition according to any one of claims 4 to 6, wherein the composition further comprises at least one immunogen.

8. The pharmaceutical composition according to any one of claims 4 to 6, wherein the composition further comprises a pharmaceutically acceptable carrier.

9. The pharmaceutical composition according to any one of claims 4 to 6, formulated for administration to an animal or human subject.

10. The pharmaceutical composition according to any one of claims 4 to 6, wherein the composition further comprises: at least one cancer therapeutic agent, wherein at least one chemotherapeutic agent and the saponin derivative are mixed or covalently linked to each other in a pharmaceutically acceptable formulation; and a pharmaceutically acceptable carrier.

11. A method of increasing the immunogenicity of an immunogen when administered to an animal or human subject, comprising the step of administering to the subject a vaccine comprising at least the pharmaceutical composition according to any one of claims 4 to 10.

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

13. The synthetic route for synthesizing saponin derivatives according to claim 12, wherein the natural saponin is obtained from Momordica cochinchinensis.

14. The synthetic route for synthesizing saponin derivatives according to claim 12, wherein the natural saponin is coupled with the functionalized side chain molecule via an amide formation reaction or an ester formation reaction.

15. A method of treating a disease, comprising: Administering a pharmaceutical composition to a subject in need thereof, wherein the pharmaceutical composition comprises a therapeutically effective amount of the composition or pharmaceutical composition according to any one of claims 1 to 10.

Images