Semi-synthetic saponin compounds

Abstract

Semi-synthetic saponin mimetic compounds and methods for use for modulating an immune system of subject and a disease, disorder, or condition are provided.

Description

SEMI-SYNTHETIC SAPONIN COMPOUNDSFEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0001] This invention was made with government support under Contract 75N93023C00042 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND

[0002] Immunomodulating compounds comprising Quillaic acid are well known and are utilized in pharmaceutical treatments. QS21, for example is a fraction of naturally-occurring saponin derived from the bark of the Qiiillaja saponaria Molina tree (Quillaja saponins). QS21 has been approved for use as an adjuvant component in several human vaccines but supply constraints of its natural source materials have limited wider use. Accordingly, there is a need for synthetic and semi-synthetic compounds with saponin-like structures for use in pharmaceutical treatments.SUMMARY

[0003] In some aspects, the presently disclosed subject matter provides a compound of formula (la) or formula (lb):

[0005] wherein:

[0006] the compound of formula (la) or formula (lb) can be in the (R) configuration, the (S) configuration, or a mixture of (R, S) configurations;

[0007] Ri is selected from H, a monosaccharide, a disaccharide, and a moiety selected from:1INIM-44840.601

[0009] each n is independently an integer selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, and 15;

[0010] R is selected from an aromatic, a saturated or unsaturated, branched or straight chain alkyl;

[0011] R2 is selected from H, a carboxylic acid, an amide, phenyl, halogen, and an aromatic;

[0012] W is O or N;

[0013] X is O, N, or S;

[0014] Y is selected from -C(O)(CH2)nCOOH, -(O)(CH2)CONH2, Ar, biaryl, branched alkyl chain, saturated alkyl chain, unsaturated alkyl chain, and halogen;

[0015] Z is selected from D-Fucose, L-Fucose, and galactose; and

[0016] pharmaceutically acceptable salts thereof.

[0019] wherein:

[0020] Ri is selected from a monosaccharide, a disaccharide, H, and a moiety selected from:R2INIM-44840.601

[0022] R is selected from an aromatic, a saturated, or unsaturated, branched or straight alkyl chain;

[0023] R2 is selected from a carboxylic acid, an amide, phenyl, halogen, an aromatic, and H;

[0024] each n is independently an integer selected from 0, 1, 2, 3, 4, 5, 6, 7. 8, 9, 10. 11. 12, 13, 14, and 15;

[0025] W is O or N;

[0026] X is O. N, or S;

[0027] Y is selected from -C(O)(CH2)nCOOH, -(O)(CH2)CONH2, Ar, biaryl, branched alkyl chain, saturated alkyl chain, unsaturated alkyl chain, and halogen;

[0028] Z is selected from D-Fucose, L-Fucose, and galactose; and

[0029] pharmaceutically acceptable salts thereof.

[0030] In certain aspects, the compound is a compound of formula (lb):

[0032] In particular aspects, the compound of formula (la) is:

[0034] wherein: R is selected from -OCH3, -OH, -NH2, -O(CH2)nCOOH, and -NH(CH2)nCOOH.

[0035] In more particular aspects, the compound of formula (la) is selected from:3INIM-44840.601

[0036] IN IM-44840.601

[0043] In more particular aspects, the compound of formula (la) is selected from:

[0044]

[0045]

[0046] 5I IM-44840.601

[0047]

[0050]

[0051] 6INIM-44840.601

[0052]

[0053]

[0054]

[0055]

[0056] In other aspects, the presently disclosed subject matter provides a method for modulating an immune system of subject, the method comprising administering a compound of formula (la) or formula (lb) to the subject.

[0057] In other aspects, the presently disclosed subject matter provides a method for treating or preventing a disease, disorder, or condition, the method comprising administering an effective amount of a compound of a compound of formula (la) or formula (lb) to a subject in need of treatment thereof.7INIM-44840.601

[0058] In certain aspects, the disease, disorder, or condition is selected from cancer, a bacterial infection, a viral infection, a fungal infection, a parasitic infection, an immune-mediated disorder, a central nervous system disease, a peripheral nervous system disease, a neurodegenerative disease, a mood disorder, a sleep disorder, a cerebrovascular disease, a peripheral artery disease, and a cardiovascular disease.

[0059] In certain aspects, the disease, disorder, or condition is cancer. In particular aspects, the cancer is selected from colorectal cancer, aero-digestive squamous cancer, lung cancer, brain cancer, liver cancer, stomach cancer, sarcoma, leukemia, lymphoma, multiple myeloma, ovarian cancer, uterine cancer, breast cancer, melanoma, prostate cancer, bladder cancer, pancreatic carcinoma, and renal carcinoma.

[0060] In certain aspects, the disease, disorder, or condition is an infection. In particular aspects, the infection comprises a bacterial, fungal, protozoal or viral infection. In more particular aspects, the bacterial, fungal, protozoal, or viral infection is selected from tuberculosis and mycobacterium avium, leprosy; pneumocystis carnii, cryptosporidiosis, histoplasmosis, toxoplasmosis, trypanosome infection, leishmaniasis, and infections caused by bacteria of the genus Escherichia, Enterobacter, Salmonella, Staphylococcus, Klebsiella, Proteus, Pseudomonas, Streptococcus, and Chlamydia, and fungal infections, including candidiasis, aspergillosis, histoplasmosis, cryptococcal meningitis, a viral disease, including respiratory syncytial virus (RSV), hepatitis B, hepatitis C, Dengue virus, herpes simplex virus, including HSV-I, HSV-II, CMV, and VZV, molluscum contagiosum, vaccinia, variola, lentivirus, human immunodeficiency virus (HIV), human papilloma virus (HPV), cytomegalovirus (CMV), varicella zoster virus (VZV), rhinovirus, enterovirus, adenovirus, coronavirus, including SARS, influenza, para-influenza, mumps virus, measles virus, papovavirus, hepadnavirus, flavivirus, retrovirus, arenavirus, including LCM, Junin virus, Machupo virus, Guanarito virus and Lassa Fever, and filovirus, including Ebola virus and Marburg virus.

[0061] In certain aspects, the method further comprises administering a therapeutic agent. In particular aspects, the therapeutic agent is selected from an immunomodulating agent, antiviral agent, an anti-inflammation agent, a chemotherapeutic agent, an anti-cancer vaccine, and hormonal therapy.

[0062] In certain aspects, the immunomodulating agent comprises a vaccine adjuvant. In certain aspects, the immunomodulating agent is selected from a liposome, alum, Freund's complete or8INIM-44840.601incomplete adjuvant, a detoxified endotoxin, and a Toll-Like Receptor agonist. Tn particular aspects, the Toll-like Receptor agonist is selected from TLR2, TLR3, TLR2 / 3, TLR4, TLR5, TLR7, TLR8, TLR7 / 8, and a TLR 9 agonist. In more particular aspects, the Toll-like Receptor agonist comprises a TLR4 agonist. In certain aspects, the TLR4 agonist is selected from natural and synthetic Lipid A derivatives and mimetics. In particular aspects, the natural and synthetic Lipid A derivatives and mimetics are selected from monophosphoryl lipid A, MPL, MPLA, 3D-MPL, GLA INI-2002, and INI-2004.

[0063] In certain aspects, the method further comprises administering an antigen. In particular aspects, the antigen is an antigen associated with Varicella zoster virus (VZV). In particular aspects, the antigen comprises an allergen. In more particular aspects, the allergen is a respiratory allergen or a food allergen. In more particular aspects, the respiratory antigen is selected from pollen, dander, and dust. In more particular aspects, the food allergen comprises a nut allergen.

[0064] Certain aspects of the presently disclosed subject matter having been stated hereinabove, which are addressed in whole or in part by the presently disclosed subject matter, other aspects will become evident as the description proceeds when taken in connection with the accompanying Examples and Drawings as best described herein below.BRIEF DESCRIPTION OF THE FIGURES

[0065] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawings will be provided by the Office upon request and payment of the necessary fee.

[0066] Having thus described the presently disclosed subject matter in general terms, reference will now be made to the accompanying Figures, which are not necessarily drawn to scale, and wherein:

[0067] FIG. 1 shows INF-y activity, a cytokine that activates, primarily targeting intracellular pathogens, promoting macrophage activation, enhancing cytotoxic T cell activity, and stimulates Thl immune responses to fight intracellular pathogens, for compounds INI-5025, INL5026, INI-5027, and INI-5028.

[0068] FIG. 2 shows IL-12p70 activity, a cytokine that regulates the immune response by stimulating T cells and producing interferon-gamma (INF-y), for compounds INI-5025, INI-5026, INI-5027, and INI-5028.9INIM-44840.601

[0069] FIG. 3 shows INF-a activity, a cytokine that activates immune cells like dendritic cells, macrophages, and T cells, and stimulates antibody class switching, for compounds INI-5025, INI-5026, INI-5027, and INI-5028.

[0070] FIG. 4 shows INF-y, a cytokine that regulates immune responses by both suppressing and enhancing certain immune cell activities, which can suppress the production of pro-inflammatory cytokines and reduce the activation of immune cells involved in autoimmune diseases, activity for compounds INI-5025, INI-5026, INI-5027, and INI-5028.

[0071] FIG. 5 shows IP- 10, chemokine that attracts activated T cells to site of inflammation, activity for compounds INI-5025, INI-5026, INI-5027, and INI-5028.

[0072] FIG. 6 shows MCP-1, a chemokine that helps recruit monocytes and macrophages to inflamed tissue, activity for compounds INI-5025, INI-5026. INI-5027, and INI-5028.

[0073] FIG. 7 shows MiP-la, a chemokine that recruits inflammatory cells to sites of infection and / or inflammation, activity for compounds INI-5025, INI-5026, INI-5027, and INI-5028.

[0074] FIG. 8 shows TNFa, which activates immune cells, neutrophils and macrophages, promoting inflammation and phagocytosis, activity for compounds INI-5025, INI-5026, INI-5027, and INI-5028.

[0075] FIG. 9 shows IL-10, a cytokine produced by macrophages and monocytes, in response to infections, injuries, or other inflammatory stimuli, activity for compounds INI-5025, INI-5026, INI-5027, and INI-5028.

[0076] FIG. 10 shows IL-6, a cytokine produced by immune cells, in response to infection, injury, or tissue damage, which stimulates the production of other inflammatory cytokines, promotes the recruitment of immune cells to the site of inflammation, and increases the expression of acutephase proteins, which help to fight infection and repair tissues, activity for compounds INI-5025, INI-5026, INI-5027, and INI-5028 and a graph of redosing with INI-5026.

[0077] FIG. 11 shows N2-specific serum antibody titers at 14 days post-secondary immunization. Statistical analysis was conducted by one-way ANOVA (GraphPad Prism). ** p>0.01, *** p>0.001.

[0078] FIG.12 shows N2-specific splenic T cell cytokine responses. Statistical analysis was conducted by one-way ANOVA (GraphPad Prism). * p>0.05, *** p>0.001, **** p>0.0001.

[0079] FIG. 13 shows N2-specific serum IgGl, IgG2 and IgG total antibody titers at 14 days postsecondary immunization.10INIM-44840.601

[0080] FIG. 14 shows N2-specific T-cell associated cytokine responses.DETAILED DESCRIPTION

[0081] The presently disclosed subject matter now will be described more fully hereinafter with reference to the accompanying Figures, in which some, but not all embodiments of the inventions are shown. Like numbers refer to like elements throughout. The presently disclosed subject matter may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Therefore, it is to be understood that the presently disclosed subject matter is not to be limited to the specific embodiments disclosed and that modifications and other embodiments are intended to be included within the scope of the appended claims.

[0082] I. COMPOSITIONS

[0083] A. Representative Compounds of Formula (la) and Formula (lb)

[0084] In some embodiments, the presently disclosed subject matter provides a compound of formula (la) or formula (lb) according to the following scheme:X = O; N; SY = C(O)(CH2)nCOOH; C(O)(CH2)nCONH2; Ar; biaryl; branched alkyl chain; saturated alkyl chain; unsaturated alkyl chain; halogens Z = D-Fucose; L- Fucose; Galactose

[0085] In some embodiments, the presently disclosed subject matter provides a compound of formula (la) or formula (lb):11INIM-44840.601

[0086]

[0087] wherein:

[0088] the compound of formula (la) or formula (lb) can be in the (R) configuration, the (S) configuration, or a mixture of (R, S) configurations;

[0089] Ri is selected from H, a monosaccharide, a disaccharide, and a moiety selected from:

[0091] each n is independently an integer selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, and 15;

[0092] R is selected from an aromatic, a saturated or unsaturated, branched or straight chain alkyl;

[0093] R2 is selected from H, a carboxylic acid, an amide, phenyl, halogen, and an aromatic;

[0094] W is O or N;

[0095] X is O, N, or S;

[0096] Y is selected from -C(O)(CH2)nCOOH, -(O)(CH2)CONH2, Ar, biaryl, branched alkyl chain, saturated alkyl chain, unsaturated alkyl chain, and halogen;

[0097] Z is selected from D-Fucose, L-Fucose, and galactose; and

[0098] pharmaceutically acceptable salts thereof.

[0099] In certain embodiments, the compound is a compound of formula (la):12INIM-44840.601(R), (S), (R. S)

[0101] wherein:

[0102] Ri is selected from a monosaccharide, a disaccharide, H, and a moiety selected from:

[0104] R is selected from an aromatic, a saturated, or unsaturated, branched or straight alkyl chain;

[0105] R2 is selected from a carboxylic acid, an amide, phenyl, halogen, an aromatic, and H;

[0106] each n is independently an integer selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, and 15;

[0107] W is O or N;

[0108] X is O, N, or S;

[0109] Y is selected from -C(O)(CH2)nCOOH, -(O)(CH2)CONH2, Ar, biaryl, branched alkyl chain, saturated alkyl chain, unsaturated alkyl chain, and halogen;

[0110] Z is selected from D-Fucose, L-Fucose, and galactose; and

[0111] pharmaceutically acceptable salts thereof.

[0112] In certain embodiments, the compound is a compound of formula (lb):13INIM-44840.601

[0114] In particular embodiments, the compound of formula (la) is:

[00115]

[0116] wherein: R is selected from -OCH3, -OH, -NH2, -O(CH2)nCOOH, and -NH(CH2)nCOOH.

[0117] In more particular embodiments, the compound of formula (la) is selected from:14INIM-44840.601

[0125] In more particular embodiments, the compound of formula (la) is selected from:15INIM-44840.601

[0126]

[0127]

[0128]

[0129]

[0130] 16INIM-44840.601

[0132]

[0133]

[00135] 17INIM-44840.601

[0136]

[00137]

[0138] Representative compounds of formula (la) and formula (lb) are provided in Table 1.Table 1. Representative Compounds of Formula (la) and Formula (lb).Compound StructureINI-5025 (R)INI 5026 (R)AKA \0-18INIM-44840.601Table 1. Representative Compounds of Formula (la) and Formula (lb).Compound StructureINI- 5027(7?, S)^- HO INI-5028 ( / ?,£)H°AP UHO INI-5029 ( / ?)PHHO^A^-O ^NNHOX-^° ^J A VN" YYHO'VINI-5030 (A)Y0y Ag $HOX*-T**X-OH0JHINI-5031 (A)nX^5V = > Y^O'~S / ^O'X^^ / ^^^- / X^X^X-X^XX<OHp Pyyy^HOXX-'^'YOY-'YYH07HINI-5032 (5) Onkyp a^-H0JH19INIM-44840.601Table 1. Representative Compounds of Formula (la) and Formula (lb).Compound StructureINI-5033 (A)y^-INI-5034 ( / ?)U-INI-5035 (A) f lRo ><S °k k°' w- - A " T-^oJHINI-5036 (R)5kkHO*X — -°XH° JHINI-5037 (7?)k k OHOA^--H0JH

[0139] Further, a structure represented generally by the formula:20INIM-44840.601- HR)n — n“(R)n

[0140] or

[0141] as used herein refers to a ring structure comprising a substituent R group, wherein the R group can be present or absent, and when present, one or more R groups can each be substituted on one or more available carbon atoms of the ring structure. The presence or absence of the R group and number of R groups is determined by the value of the variable “n,” which is an integer generally having a value ranging from 0 to the number of carbon atoms on the ring available for substitution. Each R group, if more than one, is substituted on an available carbon of the ring structure rather than on another R group. For example, the structure above where n is 2 would comprise compound groups including, but not limited to:R2

[0142]

[0143] A dashed line representing a bond in a cyclic ring structure indicates that the bond can be either present or absent in the ring. That is, a dashed line representing a bond in a cyclic ring structure indicates that the ring structure is selected from the group consisting of a saturated ring structure, a partially saturated ring structure, and an unsaturated ring structure.

[0144] The symbol ( 'WW'W' ) denotes the point of attachment of a moiety to the remainder of the molecule.

[0145] As used herein, the term “alkyl” refers to a univalent group derived from an alkane by removal of a hydrogen atom from any carbon atom resulting in a substituent group with a formula of -CnH2n+i. An alkyl group derived by removal of a hydrogen atom from a terminal carbon atom of an unbranched alkane form a subclass of normal alkyl (n- alkyl) groups having a formula - H(CH2)n. The groups RCH2, R2CH (R H), and R3C (R H) are primary, secondary and tertiary alkyl groups, respectively.

[0146] The number of carbon atoms designated in the alkyl group (i.e., C1-C10 means one to ten carbons, including 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 carbons). In particular embodiments, the term “alkyl” refers to C1-C20 inclusive, including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17,21INIM-44840.60118, 19, and 20 carbons. An alkyl group can be linear (i.e., “straight-chain”) or branched. More particularly, as used herein the term “C1-C4 alkyl” refers to an alkyl group having 1, 2, 3, or 4 carbon atoms. Representative C1-C4 alkyl groups include methyl, ethyl, w-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl. “Branched” refers to an alkyl group in which a lower alkyl group, such as methyl, ethyl or propyl, is attached to a linear alkyl chain. “Lower alkyl” refers to an alkyl group having 1 to about 8 carbon atoms (i.e.. a C1-8 alkyl), e.g., 1. 2, 3, 4, 5, 6. 7, or 8 carbon atoms.

[0147] In some embodiments, the alkyl group can be substituted. As used herein, the term “substituted alkyl” includes alkyl groups, as defined herein, in which one or more atoms of the alkyl group are replaced with another atom or functional group, including for example, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino, dialkylamino, sulfate, cyano, and mercapto.

[0148] The terms “halo,” “halide,” or “halogen” as used herein refer to fluoro, chloro, bromo, and iodo groups. In some embodiments, the halogen is F. In some embodiments, the halogen is Cl.

[0149] Throughout the specification and claims, a given chemical formula or name shall encompass all tautomers, congeners, and optical- and stereoisomers, as well as racemic mixtures where such isomers and mixtures exist.

[0150] As used herein the term "biaryl" means two aromatic rings or ring systems linked together via single bond.

[0151] Certain compounds of the present disclosure may possess asymmetric carbon atoms (optical or chiral centers) or double bonds; the enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisometric forms that may be defined, in terms of absolute stereochemistry, as (R)-or (S)- or, as D- or L- for amino acids, and individual isomers are encompassed within the scope of the present disclosure. The compounds of the present disclosure do not include those which are known in art to be too unstable to synthesize and / or isolate. The present disclosure is meant to include compounds in racemic, scalemic, and optically pure forms. Optically active (R)- and (S)-, or D- and L-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When the compounds described herein contain olefenic bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers.22INIM-44840.601

[0152] Unless otherwise stated, structures depicted herein are also meant to include all stereochemical forms of the structure; i.e., the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the disclosure.

[0153] It will be apparent to one skilled in the art that certain compounds of this disclosure may exist in tautomeric forms, all such tautomeric forms of the compounds being within the scope of the disclosure. The term “tautomer,” as used herein, refers to one of two or more structural isomers which exist in equilibrium and which are readily converted from one isomeric form to another.

[0154] Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures with the replacement of a hydrogen by a deuterium or tritium, or the replacement of a carbon by13C- or14C-enriched carbon are within the scope of this disclosure.

[0155] The compounds of the present disclosure may also contain unnatural proportions of atomic isotopes at one or more of atoms that constitute such compounds. For example, the compounds may be radiolabeled with radioactive isotopes, such as for example tritium (3H), iodine-125 (125I) or carbon- 14 (14C). All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are encompassed within the scope of the present disclosure.

[0156] In addition to salt forms, the present disclosure provides compounds, which are in a prodrug form. Prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present disclosure. Additionally, prodrugs can be converted to the compounds of the present disclosure by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of the present disclosure when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0157] The term “protecting group” refers to chemical moieties that block some or all reactive moieties of a compound and prevent such moieties from participating in chemical reactions until the protective group is removed, for example, those moieties listed and described in T. W. Greene, P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd ed. John Wiley & Sons (1999). It may be advantageous, where different protecting groups are employed, that each (different) protective group be removable by a different means. Protective groups that are cleaved under totally disparate23INIM-44840.601reaction conditions allow differential removal of such protecting groups. For example, protective groups can be removed by acid, base, and hydrogenolysis. Groups such as trityl, dimethoxytrityl, acetal and tert-butyldimethylsilyl are acid labile and may be used to protect carboxy and hydroxy reactive moieties in the presence of amino groups protected with Cbz groups, which are removable by hydrogenolysis, and Fmoc groups, which are base labile. Carboxylic acid and hydroxy reactive moieties may be blocked with base labile groups such as. without limitation, methyl, ethyl, and acetyl in the presence of amines blocked with acid labile groups such as tert-butyl carbamate or with carbamates that are both acid and base stable but hydrolytically removable.

[0158] Carboxylic acid and hydroxy reactive moieties may also be blocked with hydrolytically removable protective groups such as the benzyl group, while amine groups capable of hydrogen bonding with acids may be blocked with base labile groups such as Fmoc. Carboxylic acid reactive moieties may be blocked with oxidatively-removable protective groups such as 2,4-dimethoxybenzyl, while co-existing amino groups may be blocked with fluoride labile silyl carbamates.

[0159] Allyl blocking groups are useful in the presence of acid- and base- protecting groups since the former are stable and can be subsequently removed by metal or pi-acid catalysts. For example, an allyl-blocked carboxylic acid can be deprotected with a palladium(O)- catalyzed reaction in the presence of acid labile t-butyl carbamate or base-labile acetate amine protecting groups. Yet another form of protecting group is a resin to which a compound or intermediate may be attached. As long as the residue is attached to the resin, that functional group is blocked and cannot react. Once released from the resin, the functional group is available to react.

[0160] Typical blocking / protecting groups include, but are not limited to the following moieties:24INIM-44840.601H2C oAlloc MeCH3H3C -:CH3Teoc Boc t-butyl TBDMS

[00161] acetyl

[0162] B. Methods for Treating a Disease, Disorder, or Condition Associated with Immune Modulation

[0163] In some embodiments, the presently disclosed subject matter provides a method for modulating a subject’s immune system, the method comprising administering to the subject a compound of formula (la) and formula (lb).

[0164] As used herein, the term “contacting” refers to placement under conditions in which direct physical association occurs, including contacting of a solid with a solid, a liquid with a liquid, a liquid with a solid, or either a liquid or a solid with a cell or tissue, whether in vitro or in vivo. Contacting can occur in vitro with isolated cells or tissue or in vivo by administering to a subject.

[0165] In some embodiments, the presently disclosed compounds can be administered in combination with TLR modulators. As used herein, the term “TLR modulator” refers to an agent that is capable of either activating or downregulating one or more of the TLR pathways. A “TLR modulator” can be an agonist or an antagonist.

[0166] In certain embodiments the TLR modulator is the TLR4 agonist INI-2002 or INI-2004:25INIM-44840.601

[0167] As used herein, the term “agonist” and grammatical variations thereof refer to an agent, such as a small molecule or protein, that binds to a protein and causes, enhances, or augments (to a statistically significant degree) a particular biological effect of the protein. In certain embodiments, an agonist activates a receptor. The activation can be full, partial, or inverse. A full agonist has high efficacy, producing a full response while occupying a relatively low proportion of receptors. A partial agonist has lower efficacy than a full agonist. It produces sub-maximal activation even when occupying the total receptor population, therefore cannot produce the maximal response, irrespective of the concentration applied. An inverse agonist produces an effect opposite to that of an agonist, yet binds to the same receptor binding-site as an agonist. Agonists can be naturally occurring or artificially synthesized compounds.

[0168] In some embodiments, the presently disclosed subject matter provides a method for treating a disease, disorder, or condition associated with immune function, the method comprising administering a therapeutically effective amount of a compound of formula (la) or formula (lb) to a subject in need of treatment thereof.

[0169] In certain embodiments, the disease, disorder, or condition is selected from cancer, a bacterial infection, a viral infection, a fungal infection, a parasitic infection, an immune-mediated disorder, a central nervous system disease, a peripheral nervous system disease, a neurodegenerative disease, a mood disorder, a sleep disorder, a cerebrovascular disease, a peripheral artery disease, and a cardiovascular disease.

[0170] In particular embodiments, the disease, disorder, or condition is cancer. In more particular embodiments, the cancer is selected from colorectal cancer, aero-digestive squamous cancer, lung cancer, brain cancer, liver cancer, stomach cancer, sarcoma, leukemia, lymphoma, multiple26INIM-44840.601myeloma, ovarian cancer, uterine cancer, breast cancer, melanoma, prostate cancer, bladder cancer, pancreatic carcinoma, and renal carcinoma.

[0171] In some embodiments, the presently disclosed method further comprises administering a second therapeutic agent. In certain embodiments, the second therapeutic agent is selected from an antiviral agent, an anti-inflammation agent, a chemotherapeutic agent, an anti-cancer vaccine, and hormonal therapy.

[0172] As used herein, the term “treating” can include reversing, alleviating, inhibiting the progression of, preventing or reducing the likelihood of the disease, disorder, or condition to which such term applies, or one or more symptoms or manifestations of such disease, disorder or condition. Preventing refers to causing a disease, disorder, condition, or symptom or manifestation of such, or worsening of the severity of such, not to occur. Accordingly, the presently disclosed compounds can be administered prophylactically to prevent or reduce the incidence or recurrence of the disease, disorder, or condition.

[0173] The “subject” treated by the presently disclosed methods in their many embodiments is desirably a human subject, although it is to be understood that the methods described herein are effective with respect to all vertebrate species, which are intended to be included in the term “subject.” Accordingly, a “subject” can include a human subject for medical purposes, such as for the treatment of an existing condition or disease or the prophylactic treatment for preventing the onset of a condition or disease, or an animal subject for medical, veterinary purposes, or developmental purposes. Suitable animal subjects include mammals including, but not limited to, primates, e.g., humans, monkeys, apes, and the like; bovines, e.g., cattle, oxen, and the like; ovines, e.g.. sheep and the like; caprines, e.g., goats and the like; porcines, e.g., pigs, hogs, and the like; equines, e.g., horses, donkeys, zebras, and the like; felines, including wild and domestic cats; canines, including dogs; lagomorphs, including rabbits, hares, and the like; and rodents, including mice, rats, and the like. An animal may be a transgenic animal. In some embodiments, the subject is a human including, but not limited to, fetal, neonatal, infant, juvenile, and adult subjects. Further, a “subject” can include a patient afflicted with or suspected of being afflicted with a condition or disease. Thus, the terms “subject” and “patient” are used interchangeably herein. The term “subject” also refers to an organism, tissue, cell, or collection of cells from a subject.

[0174] In general, the “effective amount” of an active agent or drug delivery device refers to the amount necessary to elicit the desired biological response. As will be appreciated by those of27INIM-44840.601ordinary skill in this art, the effective amount of an agent or device may vary depending on such factors as the desired biological endpoint, the agent to be delivered, the makeup of the pharmaceutical composition, the target tissue, and the like.

[0175] The term “combination” is used in its broadest sense and means that a subject is administered at least two agents, more particularly a compound described herein and at least one other therapeutic agent. More particularly, the term “in combination” refers to the concomitant administration of two (or more) active agents for the treatment of a, e.g., single disease state. As used herein, the active agents may be combined and administered in a single dosage form, may be administered as separate dosage forms at the same time, or may be administered as separate dosage forms that are administered alternately or sequentially on the same or separate days. In one embodiment of the presently disclosed subject matter, the active agents are combined and administered in a single dosage form. In another embodiment, the active agents are administered in separate dosage forms (e.g., wherein it is desirable to vary the amount of one but not the other). The single dosage form may include additional active agents for the treatment of the disease state.

[0176] Further, the compounds described herein can be administered alone or in combination with adjuvants that enhance stability of the compounds, alone or in combination with one or more therapeutic agents, facilitate administration of pharmaceutical compositions containing them in certain embodiments, provide increased dissolution or dispersion, increase inhibitory activity, provide adjunct therapy, and the like, including other active ingredients. Advantageously, such combination therapies utilize lower dosages of the conventional therapeutics, thus avoiding possible toxicity and adverse side effects incurred when those agents are used as monotherapies.

[0177] The timing of administration of a compound described herein and at least one additional therapeutic agent can be varied so long as the beneficial effects of the combination of these agents are achieved. Accordingly, the phrase “in combination with” refers to the administration of a compound described herein and at least one additional therapeutic agent either simultaneously, sequentially, or a combination thereof. Therefore, a subject administered a combination of a compound described herein and at least one additional therapeutic agent can receive a compound and at least one additional therapeutic agent at the same time (i.e., simultaneously) or at different times (i.e., sequentially, in either order, on the same day or on different days), so long as the effect of the combination of both agents is achieved in the subject.28INIM-44840.601

[0178] When administered sequentially, the agents can be administered within 1, 5, 10, 30, 60, 120, 180, 240 minutes or longer of one another. In other embodiments, agents administered sequentially, can be administered within 1, 5, 10, 15, 20 or more days of one another. Where the compound described herein and at least one additional therapeutic agent are administered simultaneously, they can be administered to the subject as separate pharmaceutical compositions, each comprising either a compound or at least one additional therapeutic agent, or they can be administered to a subject as a single pharmaceutical composition comprising both agents.

[0179] When administered in combination, the effective concentration of each of the agents to elicit a particular biological response may be less than the effective concentration of each agent when administered alone, thereby allowing a reduction in the dose of one or more of the agents relative to the dose that would be needed if the agent was administered as a single agent. The effects of multiple agents may, but need not be, additive or synergistic. The agents may be administered multiple times.

[0180] In some embodiments, when administered in combination, the two or more agents can have a synergistic effect. As used herein, the terms “synergy,” “synergistic,” “synergistically” and derivations thereof, such as in a “synergistic effect” or a “synergistic combination” or a “synergistic composition” refer to circumstances under which the biological activity of a combination of a compound described herein and at least one additional therapeutic agent is greater than the sum of the biological activities of the respective agents when administered individually.

[0181] For example, synergy can be expressed in terms of a “Synergy Index (SI),” which generally can be determined by the method described by F. C. Kull et al., Applied Microbiology 9, 538 (1961), from the ratio determined by:

[0182] Qa / QA+ Qb / QB= Synergy Index (SI)

[0183] wherein:

[0184] QA is the concentration of a component A, acting alone, which produced an end point in relation to component A;

[0185] Qais the concentration of component A, in a mixture, which produced an end point;

[0186] QB is the concentration of a component B, acting alone, which produced an end point in relation to component B; and

[0187] Qb is the concentration of component B, in a mixture, which produced an end point.29INIM-44840.601

[0188] Generally, when the sum of Qa / QAand QH / QB is greater than one, antagonism is indicated. When the sum is equal to one, additivity is indicated. When the sum is less than one, synergism is demonstrated. The lower the SI, the greater the synergy shown by that particular mixture. Thus, a “synergistic combination” has an activity higher that what can be expected based on the observed activities of the individual components when used alone. Further, a “synergistically effective amount” of a component refers to the amount of the component necessary to elicit a synergistic effect in, for example, another therapeutic agent present in the composition.

[0189] C. Pharmaceutical Compositions and Administration

[0190] In another aspect, the present disclosure provides a pharmaceutical composition including one compound described herein alone or in combination with one or more additional therapeutic agents in admixture with a pharmaceutically acceptable excipient. One of skill in the art will recognize that the pharmaceutical compositions include the pharmaceutically acceptable salts of the compounds described above. Pharmaceutically acceptable salts are generally well known to those of ordinary skill in the art, and include salts of active compounds which are prepared with relatively nontoxic acids or bases, depending on the particular substituent moieties found on the compounds described herein. When compounds of the present disclosure contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent or by ion exchange, whereby one basic counterion (base) in an ionic complex is substituted for another. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salt, or a similar salt.

[0191] When compounds of the present disclosure contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent or by ion exchange, whereby one acidic counterion (acid) in an ionic complex is substituted for another. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids like hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydriodic, or phosphorous acids and the like, as well as the salts derived from relatively nontoxic organic acids like acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, methanesulfonic,30INIM-44840.601trifluoroacetic acid (TFA), and the like. Also included are salts of amino acids such as arginate and the like, and salts of organic acids like glucuronic or galactunoric acids and the like (see, for example, Berge et al, “Pharmaceutical Salts”, Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present disclosure contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0192] Accordingly, pharmaceutically acceptable salts suitable for use with the presently disclosed subject matter include, by way of example but not limitation, acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsylate, carbonate, citrate, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, mucate, napsylate, nitrate, pamoate (embonate), pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, or teoclate. Other pharmaceutically acceptable salts may be found in, for example, Remington: The Science and Practice of Pharmacy (20thed.) Lippincott, Williams & Wilkins (2000). In therapeutic and / or diagnostic applications, the compounds of the disclosure can be formulated for a variety of modes of administration, including systemic and topical or localized administration. Techniques and formulations generally may be found in Remington: The Science and Practice of Pharmacy (20thed.) Lippincott, Williams & Wilkins (2000).

[0193] Depending on the specific conditions being treated, such agents may be formulated into liquid or solid dosage forms and administered systemically or locally. The agents may be delivered, for example, in a timed- or sustained-slow release form as is known to those skilled in the art. Techniques for formulation and administration may be found in Remington: The Science and Practice of Pharmacy (20thed.) Lippincott, Williams & Wilkins (2000). Suitable routes may include oral, buccal, by inhalation spray, sublingual, rectal, transdermal, vaginal, transmucosal, nasal or intestinal administration; parenteral delivery, including intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intra-articullar, intra -sternal, intra-synovial, intra- hepatic, intralesional, intracranial, intraperitoneal, intranasal, or intraocular injections or other modes of delivery.

[0194] For injection, the agents of the disclosure may be formulated and diluted in aqueous solutions, such as in physiologically compatible buffers such as Hank’s solution, Ringer’s solution,31INIM-44840.601or physiological saline buffer. For such transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0195] Use of pharmaceutically acceptable inert carriers to formulate the compounds herein disclosed for the practice of the disclosure into dosages suitable for systemic administration is within the scope of the disclosure. With proper choice of carrier and suitable manufacturing practice, the compositions of the present disclosure, in particular, those formulated as solutions, may be administered parenterally, such as by intravenous injection. The compounds can be formulated readily using pharmaceutically acceptable carriers well known in the art into dosages suitable for oral administration. Such carriers enable the compounds of the disclosure to be formulated as tablets, pills, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject (e.g., patient) to be treated.

[0196] For nasal or inhalation delivery, the agents of the disclosure also may be formulated by methods known to those of skill in the art, and may include, for example, but not limited to, examples of solubilizing, diluting, or dispersing substances, such as saline; preservatives, such as benzyl alcohol; absorption promoters; and fluorocarbons.

[0197] In particular embodiments, the compound of formulae (I- II) is administered intranasally in a form selected from the group consisting of a nasal spray, a nasal drop, a powder, a granule, a cachet, a tablet, an aerosol, a paste, a cream, a gel, an ointment, a salve, a foam, a paste, a lotion, a cream, an oil suspension, an emulsion, a solution, a patch, and a stick.

[0198] As used herein, the term administrating via an "intranasal route" refers to administering by way of the nasal structures.

[0199] As used herein, the term "peripheral nervous system" includes the part of the nervous system comprising the nerves and ganglia on the outside of the brain and spinal cord. The peripheral nervous system connects the central nervous system to the limbs and organs and acts as a communication relay between the brain and the extremities. The presently disclosed small molecule compounds of formula (la) or formula (lb) can access the peripheral nervous system through the blood.

[0200] Intranasal administration generally allows the active agent to bypass first pass metabolism, thereby enhancing the bioavailability of the active agent. Such delivery can offer several advantages over other modes of drug delivery, including, but not limited to, increasing the onset32INIM-44840.601of action, lowering the required dosage, enhancing the efficacy, and improving the safety profile of the active agent. For example, tablet dosage forms enter the bloodstream through the gastrointestinal tract, which subjects the drug to degradation from stomach acid, bile, digestive enzymes, and other first pass metabolism effects. As a result, tablet formulations often require higher doses and generally have a delayed onset of action. Nasal administration of a drug also can facilitate compliance, especially for pediatric patients, geriatric patients, patients suffering from a neurodegenerative disease, or other patients for which swallowing is difficult, e.g., patients suffering from nausea, such as patients undergoing chemotherapy, or patients with a swallowing disorder.

[0201] Intranasal (“i.n.” or “IN”) delivery of an agent to a subject can facilitate delivery of the agent to the brain and / or peripheral nervous system. Such administration is non-invasive and offers several advantages including avoidance of hepatic first pass clearance, rapid onset of action, frequent self-administration and easy dose adjustments. Small molecules have an added advantage of being absorbed paracellularly through the nasal epithelium after which, these molecules can then directly enter the CNS through the olfactory or the trigeminal nerve associated pathway and can be directly transported to the brain upon intranasal administration.

[0202] For intranasal delivery, in addition to the active ingredients, pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. The agents of the disclosure may be formulated by methods known to those of skill in the art, and may include, for example, but not limited to, examples of solubilizing, diluting, or dispersing substances, such as saline, preservatives, such as benzyl alcohol, absorption promoters, and fluorocarbons. Optimized formulations for intranasal delivery may include addition of permeability enhancers (mucoadhesives, nanoparticles, and the like) as well as combined use with an intranasal drug delivery device (for example, one that provides controlled particle dispersion with particles aerosolized to target the upper nasal cavity).

[0203] In particular, polymer-based nanoparticles, including chitosan, maltodextrin, polyethylene glycol (PEG), polylactic acid (PLA), polylactic-co-glycolic acid (PLGA). and PAMAM dendrimer; gels, including poloxamer; and lipid-based formulations, including glycerol monocaprate (Capmul™), mixtures of mono-, di-, and triglycerides and mono- and di- fatty esters33INIM-44840.601of PEG (Labrafil™), palmitate, glycerol monostearate, and phospholipids can be used to administer the presently disclosed compounds of formula (Ia) or formula (lb) intranasally.

[0204] The presently disclosed compounds of formula (la) or formula (lb) also can be administered intranasally via mucoadhesive agents. Mucoadhesion is commonly defined as the adhesion between two materials, at least one of which is a mucosal surface. More particularly, mucoadhesion is the interaction between a mucin surface and a synthetic or natural polymer. Mucoadhesive dosage forms can be designed to enable prolonged retention at the site of application, providing a controlled rate of drug release for improved therapeutic outcome. Application of dosage forms to mucosal surfaces may be of benefit to drug molecules not amenable to the oral route, such as those that undergo acid degradation or extensive first-pass metabolism. Mucoadhesive materials suitable for use with nasal administration of the presently disclosed compounds of formula (la) or formula (lb) include, but are not limited to, soluble cellulose derivatives, such as hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), methylcellulose (MC), and carboxymethyl cellulose (CMC), and insoluble cellulose derivatives, such as ethylcellulose and microcrystalline cellulose (MCC), starch (e.g., Amioca®), polyacrylates, such as poly(acrylic acid) (e.g., Carbopol®974P), functionalized mucoadhesive polymers, such as polycarbophil, hyaluronan, and amberlite resin, and chitosan (2-amino-2-deoxy-(l— >4)-P-d- glucopyranan) formulations and derivatives thereof.

[0205] In some embodiments, the formulation also includes a permeability enhancer. As used herein, the term "permeability enhancer" refers to a substance that facilitates the delivery of a drug across mucosal tissue. The term encompasses chemical enhancers that, when applied to the mucosal tissue, render the tissue more permeable to the drug. Permeability enhancers include, but are not limited to, dimethyl sulfoxide (DMSO), hydrogen peroxide (H2O2), propylene glycol, oleic acid, cetyl alcohol, benzalkonium chloride, sodium lauryl sulphate, isopropyl myristate. Tween 80, dimethyl formamide, dimethyl acetamide, sodium lauroylsarcosinate, sorbitan monolaurate, methylsulfonylmethane, Azone, terpenes, phosphatidylcholine dependent phospholipase C, triacyl glycerol hydrolase, acid phosphatase, phospholipase A2, concentrated saline solutions (e.g., PBS and NaCl), polysorbate 80, polysorbate 20, sodium dodecanoate (C12), sodium caprate (C10) and / or sodium palmitate (C16), tert-butyl cyclohexanol (TBCH), and alpha-terpinol.

[0206] In some embodiments, the intranasal administration is accomplished via a ViaNase™ device (Kurve Technology, Inc.).34INIM-44840.601

[0207] Pharmaceutical compositions suitable for use in the present disclosure include compositions wherein the active ingredients are contained in an effective amount to achieve its intended purpose. Determination of the effective amounts is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein. Generally, the compounds according to the disclosure are effective over a wide dosage range. For example, in the treatment of adult humans, dosages from 0.01 to 1000 mg, from 0.5 to 100 mg, from 1 to 50 mg per day, and from 5 to 40 mg per day are examples of dosages that may be used. A nonlimiting dosage is 10 to 30 mg per day. The exact dosage will depend upon the route of administration, the form in which the compound is administered, the subject to be treated, the body weight of the subject to be treated, the bioavailability of the compound(s), the adsorption, distribution, metabolism, and excretion (ADME) toxicity of the compound(s), and the preference and experience of the attending physician.

[0208] In addition to the active ingredients, these pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. The preparations formulated for oral administration may be in the form of tablets, dragees, capsules, or solutions.

[0209] Pharmaceutical preparations for oral use can be obtained by combining the active compounds with solid excipients, optionally grinding a resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethyl-cellulose (CMC), and / or polyvinylpyrrolidone (PVP: povidone). If desired, disintegrating agents may be added, such as the cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.

[0210] Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol (PEG), and / or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dye-stuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active compound doses.35INIM-44840.601

[0211] Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin, and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and / or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols (PEGs). In addition, stabilizers may be added.

[0212] Following long-standing patent law convention, the terms “a,” “an,” and “the” refer to “one or more” when used in this application, including the claims. Thus, for example, reference to “a subject” includes a plurality of subjects, unless the context clearly is to the contrary (e.g., a plurality of subjects), and so forth.

[0213] Throughout this specification and the claims, the terms “comprise,” “comprises,” and “comprising” are used in a non-exclusive sense, except where the context requires otherwise. Likewise, the term “include” and its grammatical variants are intended to be non-limiting, such that recitation of items in a list is not to the exclusion of other like items that can be substituted or added to the listed items.

[0214] For the purposes of this specification and appended claims, unless otherwise indicated, all numbers expressing amounts, sizes, dimensions, proportions, shapes, formulations, parameters, percentages, quantities, characteristics, and other numerical values used in the specification and claims, are to be understood as being modified in all instances by the term “about” even though the term “about” may not expressly appear with the value, amount or range. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are not and need not be exact, but may be approximate and / or larger or smaller as desired, reflecting tolerances, conversion factors, rounding off, measurement error and the like, and other factors known to those of skill in the art depending on the desired properties sought to be obtained by the presently disclosed subject matter.

[0215] The term “about,” when used in connection with one or more numbers or numerical ranges, should be understood to refer to all such numbers, including all numbers in a range and modifies that range by extending the boundaries slightly above and slightly below the numerical values set forth by, for example, in some embodiments, + / -20%, + / - 15%, + / -10%, + / -5%. + / -4%, + / -3%, + / -2%, and + / -1%. The recitation of numerical ranges by endpoints includes all numbers, e.g., whole36INIM-44840.601integers, including fractions thereof, subsumed within that range (for example, the recitation of 1 to 5 includes 1, 2, 3, 4, and 5, as well as fractions thereof, e.g., 1.5, 2.25, 3.75, 4.1, and the like) and any range within that range.

[0216] Further, the term “about” when used in connection with one or more numbers or numerical ranges, should be understood to refer to all such numbers, including all numbers in a range and modifies that range by extending the boundaries above and below the numerical values set forth. The recitation of numerical ranges by endpoints includes all numbers, e.g., whole integers, including fractions thereof, subsumed within that range (for example, the recitation of 1 to 5 includes 1, 2, 3, 4, and 5, as well as fractions thereof, e.g., 1.5, 2.25, 3.75, 4.1, and the like) and any range within that range.EXAMPLES

[0217] The following Examples have been included to provide guidance to one of ordinary skill in the art for practicing representative embodiments of the presently disclosed subject matter. In light of the present disclosure and the general level of skill in the art, those of skill can appreciate that the following Examples are intended to be exemplary only and that numerous changes, modifications, and alterations can be employed without departing from the scope of the presently disclosed subject matter. The synthetic descriptions and specific examples that follow are only intended for the purposes of illustration, and are not to be construed as limiting in any manner to make compounds of the disclosure by other methods.37INIM-44840.601EXAMPLE 1Synthetic Schemes and Experimental Data

[0218] Scheme 1: Synthesis of disaccharide donor.

[0219] Scheme 2: Synthesis of functionalized D-fucose.

[0220] Scheme 3: Synthesis of fully deprotected trisaccharide.

[0221] Scheme 4: Synthesis of lipid-glycerol conjugate.38INIM-44840.601

[0222] Scheme 5: Quillaic acid and coupling with lipid-glycerol conjugate.

[0223] Scheme 6: Synthesis of fully deprotected semi-synthetic saponins.39INIM-44840.601

[0224] Scheme 7: Synthesis of glucuronic acid (INI-5029) analog of INI-5026.

[0225] Scheme 8: Synthesis of INI-5034.40INIM-44840.601

[0226] Scheme 9: Synthesis of lipidated derivative (INI-5036) of INI-5026.

[0227] Scheme 10: Synthesis of INI-5026 derivatives with varying acyl chain length, including INI-5037 (n = 2), INI-5026 (n = 5), and INI-5032 (n = 8).41INIM-44840.601

[0228] Scheme 11: Synthesis of ether analog (INI-5035) of INI-5026.Scheme 12: Synthesis of galactose analog (INI-5030) of INI-5026.42INIM-44840.601

[0229] Synthesis of (2S,3 R,4R, 5S, 6S )-6-methyltetrahydro-2H-pyran-2, 3, 4, 5 -tetrayl tetraacetate (4): To a stirring solution of L- Rhamnose (1) in acetic anhydride (25 mL) in ice-cooled temperature, pyridine (20 eq.) was added and allowed to stir at room temperature for 6 hours until the complete consumption of 1 as confirmed by TLC (EtOAc / Hep; 1:1). The crude mixture was concentrated under reduced pressure and dissolved in ethyl acetate (100 mL) and washed with IN HC1 (50 mL), brine (50 mL) and dried over Na2SC>4 and concentrated under reduced pressure. The residue was dried in high vacuum for 2 hours to afford 4 (95%) as a colorless liquid. Rf = 0.6 (EtOAc / Hep; 1:2).

[0230] Synthesis of (2S,3S,4R, 5R, 6S )-2-methyl-6-( p-tolylthio )tetrahydro-2H-pyran-3,4,5-triyl triacetate (5): To a stirred solution of 4 in anhydrous DCM was added 4-methylbenzenethiol (1.1 eq.) at 0 °C and allowed to stir for 15 minutes. BFs-EtzO (1.1 eq.) was added to this solution dropwise and allowed to stir at this temperature. Upon the completion of reaction after 6 hours, the reaction was quenched by addition of TEA (1 mL) at 0 °C. The crude mixture was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 4 to 1 / 2) to afford 5 (65%) as a white solid: Rf = 0.4 (EtOAc / Hep=l / 2).

[0231] Synthes is of (2S,3R,4R,5R,6S)-2-methyl-6-(p-tolylthio)tetrahydro-2H-pyran-3,4,5-triol (6): To a stirred solution of 5 in methanol was added K2CO3 (4 eq.) and allowed to stir at room temperature for 2 hours. After the completion of reaction, the reaction was concentrated under reduced pressure, acidified with IN HC1 (50 mL) and extracted with EtOAc (100 mL), washed with brine (100 mL) and dried over N 2SC>4. The organic layer was concentrated under reduced pressure and dried in high vacuum for 2 hours to afford 6 (65%) as a white foam: Rf = 0.3 (DCM / MeOH=9 / l).

[0232] Synthesis of ( 3aR, 4S,6S, 7S, 7aR )-2,2, 6-trimethyl-4-( p-tolylthio )tetrahydro-4H- [l,3]dioxolo[4,5-c]pyran-7-ol (7): To a stirred suspension of 6 in 50 mL of DCM / Acetone (3 / 1) was added 2,2 dimethoxy propane (20 eq.) and p-Toluenesulfonic acid (0.1 eq.). After stirring for 5 hours at room temperature, the reaction was quenched by adding sat. NaHCCh and extracted with DCM (100 mL), washed with brine (100 mL) and dried over Na2SC>4. The crude mixture was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 4 to 1 / 2) to afford 7 (65%) as a white solid: Rf = 0.4 (EtOAc / Hep=l / 2).

[0233] Synthesis of protected xylose (8): To a stirring solution of Xylose (2) in acetic anhydride (25 mL) in ice-cooled temperature, pyridine (20 eq.) was added and allowed to stir at room43INIM-44840.601temperature for 6 hours until the complete consumption of 2 as confirmed by TLC (EtoAC / Hep=l / l). The crude mixture was concentrated under reduced pressure and dissolved in ethyl acetate (100 mL) and washed with IN HC1 (50 mL), brine (50 mb) and dried over Na2SC>4 and concentrated under reduced pressure. The residue was dried in high vacuum for 2 hours to afford 8 (95%) as a colorless liquid. Rf = 0.6 (EtOAc / Hep=l / 2).

[0234] Synthesis of protected xylose tricloroacetamide (9): To a stirred solution of 8 in DMF (100 mL) was added hydrazine acetate (1.2 eq.) and allowed to stir at room temperature for 12 hours. After the completion of reaction, it was quenched by adding ice cooled water, extracted with EtOAc (250 mL), washed with brine (100 mL) and dried over Na2SC>4. The crude mixture was then concentrated and used for next step without purification. Rf = 0.4 (EtOAc / Hep=l / l). The crude concentrate was then dissolved in anhydrous DCM (200 mL) and allowed to stir for 5 minutes. Trichloro acetonitrile (1.2 eq.) and DBU (1.1 eq.) was then added to the reaction mixture subsequently and allowed to stir for 6 hours until the complete consumption of starting material. The reaction mixture was concentrated and quickly purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 9 (65%) as a colorless liquid: Rf = 0.6 (EtOAc / Hep=l / 2).

[0235] Synthesis of protected Di saccharides (10): To a stirred solution of 7 in anhydrous DCM (200 mL) was added 9 (1.1 eq.) and cooled down to -78 °C and allowed to stir for 15 minutes. BF3-Et2O (1.1 eq.) was added to this solution dropwise and allowed to stir at this temperature. Upon the completion of reaction after 1 hour, the reaction was quenched by addition of TEA (1 mL) at -30 °C. The crude mixture was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 10 (55%) as a white solid: Rf = 0.4 (EtOAc / Hep=l / 2).

[0236] Synthesis of (3R,4S,5S,6R)-6-methyltetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate (11):To a stirring solution of D-fucose (3) in acetic anhydride (25 mL) in ice-cooled temperature, pyridine (20 eq.) was added and allowed to stir at room temperature for 6 hours until the complete consumption of 3 as confirmed by TLC (EtoAC / Hep=l / l). The crude mixture was concentrated under reduced pressure and dissolved in ethyl acetate (100 mL) and washed with IN HC1 (50 mL), brine (50 mL) and dried over Na2SO4 and concentrated under reduced pressure. The residue was dried in high vacuum for 2 hours to afford 11 (95%) as a colorless liquid. Rf = 0.6 (EtOAc / Hep=l / 2).44INIM-44840.601

[0237] Synthesis of (2R,3R,4S,5S,6R)-2-(2-hromoethoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyl triacetate (12): To a stirred solution of 11 in anhydrous DCM at 0 °C was added bromoethanol (1.1 eq.) and allowed to stir for 15 minutes. BF3-Et2O (1.1 eq.) was added to this solution dropwise and allowed to stir at this temperature. Upon the completion of reaction after 3 hours, the reaction was quenched by addition of TEA (1 mL) at 0 °C. The crude mixture was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 4 to 1 / 2) to afford 12 (65%) as a colorless foam: Rf = 0.4 (EtOAc / Hep=l / l).

[0238] Synthesis of(2R,3R,4S,5S,6R)-2-(2-azidoethoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyl triacetate (13): To a stirred solution of 12 in DMSO (100 mL) was added Na? (1.1 eq.) slowly portion wise and allowed to stir at room temperature for 5 hours until the complete consumption of 12 and formation of 13 as identified from TLC and LC-MS. The crude reaction mixture was quenched by adding ice cooled water and extracted with EtOAc (250 mL), washed with brine (100 mL) and dried over NazSCL. The organic layer was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 2 / 10 to 5 / 10) to afford 13 (55%) as a white solid. Rf = 0.4 (EtOAc / Hep=l / l).

[0239] Synthesis of ( 3aS,4R,6R, 7R, 7aR )-6-(2-azidoethoxy)-2,2,4-trimethyltetrahydro-4H-[ 1,3 / dioxolol 4,5-c lpyran-7-ol (14): To a stirred suspension of 13 in methanol (100 mL) was added K2CO3 (4.5 eq.) at room temperature and allowed to stir for 2 hours. After the completion of reaction, the crude mixture was filtered, concentrated and dried over high vacuum for 5 hours and used for next step without further purification. The crude concentrate was then suspended in 100 mL of DCM / Acetone (3 / 1). 2,2 dimethoxy propane (20 eq.) and p-Toluenesulfonic acid (0.1 eq.) was then added subsequently. After stirring for 5 hours at room temperature, the reaction was quenched by adding sat. NaHCCL and extracted with DCM (200 mL), washed with brine (100 mL) and dried over Na2SC>4. The crude mixture was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 4 to 1 / 2) to afford 14 (65%) as an off-white solid: Rf - 0.5 (EtOAc / Hep=2 / 3).

[0240] Synthesis of protected trisaccharide (15): The stirred solution of 14 and 10 (1.1 eq.) in anhydrous DCM (150 mL) was cooled down to -78 °C and allowed to stir for 15 minutes. BF3. Et2O (1.1 eq.) and NIS (1.1 eq.) was added to this solution and allowed to stir at this temperature. Upon the completion of reaction after 1 hours, the reaction was quenched by addition of TEA (1 mL) at45INIM-44840.601-30 °C. The crude mixture was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 15 (55%) as a white solid: Rf = 0.5 (EtOAc / Hep=l / 2).

[0241] Synthesis of protected trisaccharide (18): The stirred solution of 23 and 10 (1.1 eq.) in anhydrous DCM (150 mL) was cooled down to -78 °C and allowed to stir for 15 minutes. BF3. Et2O (1.1 eq.) and NIS (1.1 eq.) was added to this solution and allowed to stir at this temperature. Upon the completion of reaction after 1 hour, the reaction was quenched by addition of TEA (1 mL) at -30 °C. The crude mixture was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 2 / 5) to afford 18 (60%) as a white solid: Rf = 0.6 (EtOAc / Hep=l / l).

[0242] Synthesis of trisaccharide (16): A stirred solution of 15 in 90% acetic acid (100 mL) was heated at 70 °C for 3 hours until the complete deprotection of di-actinides. The crude mixture was concentrated under reduced pressure and dried over high vacuum for 3 hours. The crude concentrate was then dissolved in methanol and K2CO3 (4.5 eq.) was added to it. The reaction was completed in 2 hours, and the solvent was evaporated under reduced pressure. The concentrate was then purified by column chromatography (silica gel; MeOH / DCM = 1 / 100 to 1 / 10) to afford 16 (55% for 2 steps) as a white solid: Rf = 0.5 (MeOH / DCM=l / 10).

[0243] Synthesis of trisaccharide (19): A stirred solution of 18 in 90% acetic acid (100 mL) was heated at 70 °C for 3 hours until the complete deprotection of di-actinides. The crude mixture was concentrated under reduced pressure and dried over high vacuum for 3 hours. The crude concentrate was then dissolved in methanol and K2CO3 (4.5 eq.) was added to it. The reaction was completed in 2 hours, and the solvent was evaporated under reduced pressure. The concentrate was then purified by column chromatography (silica gel; MeOH / DCM = 1 / 100 to 1 / 10) to afford 19 (55% for 2 steps) as a white solid: Rf = 0.5 (MeOH / DCM=l / 10).

[0244] Synthesis of (R)-tert-butyl((2,2-dimethyl-l,3-dioxolan-4-yl)methoxy)diphenylsilane (21 ):To a stirred solution of 20 in DMF (100 mL) was added ter / -Butyl(chloro)diphenylsilane (1.2 eq.) and Imidazole (1.2 eq.) at room temperature. The reaction was allowed to stir for 12 hours and quenched by adding ice cooled water, extracted with EtOAC (250 mL), washed with brine (100 mL) and dried over NaiSCL. The organic layer was concentrated under reduced pressure and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 1 / 5) to afford 21 (85%) as a white solid: Rf = 0.7 (EtOAc / Hep=l / 5).

[0245] Synthesis of (R, S)-tert-butyl((2,2-dimethyl-l,3-dioxolan-4-yl)methoxy)diphenylsilane (28):To a stirred solution of 27 in DMF (100 mL) was added tert-Butyl(chloro)diphenylsilane (1.2 eq.)46INIM-44840.601and Imidazole (1.2 eq.) at room temperature. The reaction was allowed to stir for 12 hours and quenched by adding ice cooled water, extracted with EtOAc (250 mL), washed with brine (100 mb) and dried over Na2SC>4. The organic layer was concentrated under reduced pressure and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 1 / 5) to afford 28 (85%) as a white solid: Rt = 0.7 (EtOAc / Hep=l / 5).

[0246] Synthesis of (R)-3-((tert-butyldiphenylsilyl)oxy)propane-l,2-diol (22): Compound 21 was dissolved in 90% acetic acid in water (100 mL) and allowed to stir for 30 minutes at 80 °C. After the completion of reaction, the crude mixture was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 2 / 5) to afford 22 (55%) as an off-white solid: Rf= 0.5 (EtOAc / Hep=l / l).

[0247] Synthesis of (R, S)-3-((tert-butyldiphenylsilyl)oxy)propane-l,2-diol (22): Compound 28 was dissolved in 90% acetic acid in water (100 mL) and allowed to stir for 30 minutes at 80 °C. After the completion of reaction, the crude mixture was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 2 / 5) to afford 29 (55%) as an off-white solid: Rf= 0.5 (EtOAc / Hep=l / l).

[0248] Synthesis of (R)-l-( tert-butyl) 12-( 3-( ( tert-butyldiphenylsilyl)oxy)-2-hydroxypropyl) dodecanedioate (24): To a stirred solution of 22 in anhydrous DCM (200 mL) at 0 °C was added EDC methiodide (1.2 eq.) and DMAP (0.1 eq.) and allowed to stir for 15 minutes. Tert-butoxy dodecanoic acid (1.1 eq.) was then added to the reaction mixture portion wise and allowed to stir at room temperature for 2 hours. After the completion of the reaction, it was quenched by adding water, extracted with DCM (200 mL), washed with brine (100 mL) and dried over Na2SC>4. The organic layer was then concentrated under reduced pressure and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 24 (55%) as a white solid: Rf = 0.5 (EtOAc / Hep=l / 2).

[0249] Synthesis of (R, S)-1 -(tert-butyl) 12-(3-((tert-butyldiphenylsilyl)oxy)-2-hydroxypropyl) dodecanedioate (30): To a stirred solution of 29 in anhydrous DCM (200 mL) at 0 °C was added EDC methiodide (1.2 eq.) and DMAP (0.1 eq.) and allowed to stir for 15 minutes. Tert-butoxy dodecanoic acid (1.1 eq.) was then added to the reaction mixture portion wise and allowed to stir at room temperature for 2 hours. After the completion of the reaction, it was quenched by adding water, extracted with DCM (200 mL), washed with brine (100 mL) and dried over Na2SO4. The organic layer was then concentrated under reduced pressure and purified by column47INIM-44840.601chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 30 (55%) as a white solid: Rf = 0.5 (EtOAc / Hep=l / 2).

[0250] Synthesis of (R)-l-( tert-butyl) 12-(3-( ( tert-butyldiphenylsilyl)oxy)-2-(prop-2-yn-l-yloxy)propyl) dodecanedioate (25): To a stirred solution of 24 in DMF (100 mL) at 0 °C was added NaH (1.2 eq.) and allowed to stir for 30 minutes. Propargyl bromide (1.1 eq.) was then added dropwise into the reaction mixture and allowed to stir for 3 hours at room temperature. The reaction was quenched by adding ice cooled water and extracted with EtOAc (200 mL), washed with brine (100 mL) and dried over NazSCU. The organic layer was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 25 (55%) as a white solid: Rf = 0.5 (EtOAc / Hep=l / l).

[0251] Synthesis of (R)-l -(tert-butyl) 12-(3-((tert-butyldiphenylsilyl)oxy)-2-(prop-2-yn-l-yloxy)propyl) dodecanedioate (31 ): To a stirred solution of 30 in DMF (100 mL) at 0 °C was added NaH (1.2 eq.) and allowed to stir for 30 minutes. Propargyl bromide (1.1 eq.) was then added dropwise into the reaction mixture and allowed to stir for 3 hours at room temperature. The reaction was quenched by adding ice cooled water and extracted with EtOAc (200 mL), washed with brine (100 mL) and dried over NazSO4. The organic layer was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 31 (55%) as a white solid: Rf = 0.5 (EtOAc / Hep=l / l).

[0252] Synthesis of(R)-l-( 3-bromo-2-( rop-2-yn-l -yloxy)propyl) 12-( tert-butyl) dodecanedioate (26): To a stirred solution of 25 in THF (100 mL) was added IM TBAF (0.5 eq.) and allowed to stir for 6 hours until the complete deprotection of silyl group. The reaction mixture was concentrated under reduced pressure and dried in high vacuum for 3 hours. The crude concentrate was then dissolved in DCM (150 mL) and CBr4 (2.5 eq.) and TPP (2 eq.) were subsequently added to it. The reaction was stirred for 1 until the formation of 26 as dictated by TLC. The crude mixture was then concentrated under reduced pressure and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 4 to 1 / 2) to afford 26 (60%) as a colorless liquid: Rf = 0.4 (EtOAc / Hep=l / 15).

[0253] Synthesis of (R, S)-l-(3-bromo-2-(prop-2-yn-l-yloxy)propyl) 12-(lerl-bulyl) dodecanedioate (32): To a stirred solution of 31 in THF (100 mL) was added IM TBAF (0.5 eq.) and allowed to stir for 6 hours until the complete deprotection of silyl group. The reaction mixture was concentrated under reduced pressure and dried in high vacuum for 3 hours. The crude concentrate was then dissolved in DCM (150 mL) and CBr4 (2.5 eq.) and TPP (2 eq.) were48INIM-44840.601subsequently added to it. The reaction was stirred for 1 hour until the formation of 26 as dictated by TLC. The crude mixture was then concentrated under reduced pressure and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 4 to 1 / 2) to afford 32 (60%) as a colorless liquid: Rf= 0.4 (EtOAc / Hep=l / 15).

[0254] Extraction of Quillaic acid (33): Saponin was subjected to hydrolytic conditions to generate the corresponding quillaic acid. The reaction mixture was processed using conventional workup procedures, and the resulting product was isolated and purified using standard purification techniques (silica gel; EtOAc / Heptane = 1 / 6 to 2 / 1) to afford 33 (2%) as an off-white solid: Rf = 0.4 (EtOAc / DCM=l / l).

[0255] Synthesis of 1 -(tert-butyl) 12-((lR)-2-(((4aR,5R,6aS,6bR,8aR,9S,10S,12aR,14bR)-9-formyl-5, 10-dihydroxy-2,2, 6a, 6b, 9,12a-hexamethyl- l,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-l- (prop-2-yn-l-yloxy)ethyl) dodecanedioate (34): To a stirred solution of 33 in DMF (100 mL) was added CS2CO3 (1.2 eq.) and allowed to stir at 85 °C for 15 min. 26 (1.1 eq.) was then added to it and allowed to stir at this temperature for next 1 hour until the complete conversion of 33. The reaction mixture was then allowed to cool down to room temperature, filtered, poured with ice cooled water and extracted with EtOAc (250 mL). The organic layer was washed with brine (100 mL), dried over Na2SC>4 and concentrated under reduced pressure. The concentrated crude was then purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 6 to 1 / 3) to afford 34 (55%) as white solid: Rf = 0.4 (EtOAc / Hep=l / l).

[0256] Synthesis of 1 -( tert-butyl ) 12-(( lR, S)-2 -((( 4aR, 5R, 6aS,6bR, 8aR, 9S, 10S,12aR,14bR )-9-formyl-5, 10-dihydroxy-2,2, 6a, 6b, 9,12a-hexamethyl- l,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-l- (prop-2-yn-l-yloxy)ethyl) dodecanedioate (36): To a stirred solution of 33 in DMF (100 mL) was added CS2CO3 (1.2 eq.) and allowed to stir at 85 °C for 15 min. 32 (1.1 eq.) was then added to it and allowed to stir at this temperature for next 1 hour until the complete conversion of 33. The reaction mixture was then allowed to cool down to room temperature, filtered, poured with ice cooled water and extracted with EtOAc (250 mL). The organic layer was washed with brine (100 mL), dried over Na2SO4 and concentrated under reduced pressure. The concentrated crude was then purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 6 to 1 / 3) to afford 36 (55%) as white solid: Rf = 0.4 (EtOAc / Hep=l / l).49INIM-44840.601

[0257] Synthesis of 12-( ((1R)-2-(((4aR,5R,6aS,6bR,8aR,9S,10S,12aR,14bR)-9-formyl-5,10-dihydroxy-2,2,6a,6b,9,12a-hexamethyl-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-1-(prop-2-yn-1-yloxy)ethoxy)-12-oxododecanoic acid (35):To a stirred solution of 34 in DCM (10 mL) was added TFA (2 mL) and allowed to stir for 2 hours at room temperature until the complete deprotection of tert-butyl group. The crude mixture was then concentrated and purified by column chromatography (silica gel: EtOAc / hexanes = 1 / 6 to 4 / 1) to afford 35 (30%) as white foam: Rf = 0.4 (EtOAc / Hep-2 / l ).

[0258] Synthesis of 12-( ( lR, S)-2-( ( ( 4aR, 5R, 6aS,6bR, 8aR, 9S, 10S,12aR, 14bR )-9-formyl-5,10-dihydroxy-2,2,6a,6b,9,12a-hexaniethyl-l,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl )oxy)-l -( prop-2 -yn-1 -yloxy)ethoxy )-12-oxododecanoic acid (37): To a stirred solution of 36 in DCM (10 mL) was added TFA (2 mL) and allowed to stir for 2 hours at room temperature until the complete deprotection of tert-butyl group. The crude mixture was then concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 6 to 4 / 1) to afford 37 (30%) as white foam: Rf = 0.4 (EtOAc / Hep=2 / l).

[0259] Synthesis of INI-5025: To a stirred solution of 19 and 35 (1.1 eq.) in 100 mL of dioxane / water (5:1) was added sodium ascorbate (0.3 eq.) and CuSO4 (0.15 eq.) and allowed to stir at room temperature for 12 hours. After complete consumption of 19, the reaction mixture was filtered, concentrated and purified by column chromatography (silica gel; Methanol / DCM = 1 / 10 to 3 / 10) to afford INI-5025 (40%) as white solid. Rf = 0.4 (MeOH / DCM=3 / 10).

[0260] 5,y / 7t / ?e. A of INI-5026: To a stirred solution of 16 and 35 (1.1 eq.) in 100 mL of dioxane / water (5:1) was added sodium ascorbate (0.3 eq.) and CuSCL (0.15 eq.) and allowed to stir at room temperature for 12 hours. After complete consumption of 16, the reaction mixture was filtered, concentrated and purified by column chromatography (silica gel; Methanol / DCM = 1 / 10 to 3 / 10) to afford INI-5026 (40%) as white solid. Rf = 0.3 (MeOH / DCM=2 / 5).

[0261] Synthesis of INI-5027: To a stirred solution of 16 and 37 (1.1 eq.) in 100 mL of dioxane / water (5:1) was added sodium ascorbate (0.3 eq.) and CuSO4 (0.15 eq.) and allowed to stir at room temperature for 12 hours. After complete consumption of 16, the reaction mixture was filtered, concentrated and purified by column chromatography (silica gel; Methanol / DCM = 1 / 10 to 3 / 10) to afford INI-5027 (40%) as white solid. Rf = 0.3 (MeOH / DCM=2 / 5).

[0262] Synthesis of INI-5028: To a stirred solution of 19 and 37 (1.1 eq.) in 100 mL of dioxane / water (5:1) was added sodium ascorbate (0.3 eq.) and CuSO4 (0.15 eq.) and allowed to50INIM-44840.601stir at room temperature for 12 hours. After complete consumption of 19, the reaction mixture was filtered, concentrated and purified by column chromatography (silica gel; Methanol / DCM = 1 / 10 to 3 / 10) to afford INI-5028 (40%) as white solid. Rf = 0.4 (MeOH / DCM=3 / 10).

[0263] Synthesis of (2S, 3R, 4S,5S, 6S )-6-( methoxy carbonyl)tetrahydro-2H-pyran-2, 3,4,5-tetrayl tetraacetate (38): To a stirred solution of D-Glucurono-6,3-lactone in methanol was added NaOMe (0.1 equiv.) and allowed to stir at room temperature for 2 hours. After the completion of reaction, the reaction was concentrated under reduced pressure, acidified with IN HC1 (50 mL) and extracted with EtOAc (100 mL), washed with brine (100 mL) and dried over NaiSCL. The organic layer was concentrated under reduced pressure and dried in high vacuum for 2 hours. To this crude intermediate pyridine (20 equiv.) and acetic anhydride was added and allowed to stir at room temperature for 5 hours until the complete consumption of intermediate as confirmed by TLC (EtOAc / Hep=l / 2). The crude mixture was concentrated under reduced pressure and dissolved in DCM (100 mL) and washed with water (100 mL) and Brine (50 mL) and dried over NaiSCL. The crude product was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 4 to 1 / 2) to afford 2 (80%) over 2 steps as a white solid: Rf = 0.3 Rf = 0.4 (EtOAc / Hep=l / 2).

[0264] Synthesis of (3R,4S,5S,6S)-2-hydroxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (39): To a stirred solution of 38 in DMF (100 mL) was added Hydrazine acetate (1.1 equiv.) and allowed to stir at room temperature for 3 hours until the complete conversion of 2 into hemiacetal as confirmed by TLC (EtOAc / Hep=l / l). The crude reaction mixture was quenched by adding sat. NaHCOs (100 mL) and extracted with EtOAc, washed with brine and dried over Na2SO4. The organic layer was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 2 to 2 / 1) to afford 3 (55%) as a white solid (anomeric mixture): Rf = 0.4 (EtOAc / Hep=l / l).

[0265] Synthesis of 40: The anomeric mixture (39) was dissolved in anhydrous DCM (200 mL) and allowed to stir for 5 minutes. Trichloro acetonitrile (1.2 equiv.) and DBU (1.1 equiv.) was then added to the reaction mixture subsequently and allowed to stir for 3 hours until the complete consumption of starting material. The reaction mixture was concentrated and quickly purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 40 (65%) as a colorless liquid: Rf = 0.6 (EtOAc / Hep=l / 2).51INIM-44840.601

[0266] Synthesis of (2R,3R,4S,5S,6S)-2-(((3S,4S,4aR,6aR,6bS,8R,8aR, 12aR, 14bR)-8a- ( ( allyloxy )carbonyl)-4-formyl-8-hydroxy-4, 6a, 6b,ll,ll, 14b-hexamethyl- 1,2, 3, 4, 4a, 5, 6,6a, 6b, 7, 8,8a, 9,10,11,12,12a, 14,14a,14b-icosahydropicen-3-yl )oxy )-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyl triacetate (42): To a stirred solution of 40 (1.2 equiv.) and 41 (1 equiv.) in anhydrous DCM (50 mL) was added B(PhFs)3 and allowed to stir at room temperature for 1 hour until the complete consumption of 41 as confirmed by TLC (EtOAc / Hep-l / l) and LCMS. The crude mixture was quenched by addition of TEA (1.2 equiv.) and concentrated. It was purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 6 (30%) as a white solid (0 anomer): Rf = 0.6 (EtOAc / Hep=l / 2).

[0267] Synthesis of compound 42a: To a stirred solution of 42 in ethanol (50 mL) was added IN NaOH (25 mL) and allowed to stir at room temperature for 2 hours and quenched by the addition of IM HC1 and lower the pH around 5. The crude mixture was extracted by EtOAc, washed with brine and dried over Na SCL. The organic phase was concentrated to afford 42a (90% crude) as off white solid without further purification.

[0268] Synthesis of compound 42b: The crude acid 42a was dissolved in DMF (50 mL) and potassium carbonate (1.5 equiv.) was added and stirred for 10 min. Benzyl bromide (1.1 equiv.) was added to the crude mixture and allowed to stir for 8 hours at room temperature. The reaction mixture was quenched by adding ice cooled water (50 mL). It was extracted with EtOAc (100 mL) and washed with brine and dried over NazSO4 and concentrated to afford 42b (65% crude) as a colorless oil without further purification.

[0269] Synthesis of benzyl (2S,3R,4S,5R,6R)-6-(((3S,4aR,6aR,6bS,8R,8aR,12aR,14bR)-8a-((allyloxy)carbonyl)-4,4,6a.6b, 11,11,14b-heptamethyl-8-((triethylsilyl)oxy)-l,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-3,4,5-tris((triethylsilyl)oxy)tetrahydro-2H-pyran-2-carboxylate (43): To a stirred solution of 42b in DMF (20 mL) was added 2,6 Lutidine and stirred for 10 min. TES Triflate was then added to the reaction mixture drop wise and allowed to stir for 6 hours at room temperature. The reaction mixture was quenched by the addition of H2O, extracted with DCM, washed with brine, dried over Na2SOr, and then concentrated under reduced pressure. The residue was purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 43 (60%) as a white solid: Rf = 0.6 (EtOAc / Hep=l / 2).52INIM-44840.601

[0270] Synthesis of (4aR,5R,6aS,6bR,8aR,9S,10S,12aRJ4bR)-10-(((2R,3R,4S,5R,6S)-6- ( ( benzyloxy)carbonyl)-3,4, 5-tris( ( triethylsilyl )oxy)tetrahydro-2H-pyran-2-yl )oxy)-9-formyl-2,2, 6a, 6b, 9,12a-hexamethyl-5-( ( triethylsilyl )oxy)-l,3,4,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylic acid (44): To a stirred solution of 43 in dioxane (10 mL) was added formic acid (25 equiv.), Triphenylphosphine (3 equiv.). Triethylamine (24 equiv.), Palladium acetate (1 equiv.) and stirred for 12 hours at room temperature. After the completion of reaction as confirmed by TLC and LCMS, the crude mixture was filtered, concentrated under reduced pressure and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 9 (60%) as a white solid: Rf= 0.6 (EtOAc / Hep=l / 2).

[0271] Synthesis of 1-benzyl 12-((lS)-2-(((4aR,5R,6aS,6bR,8aR,9S,10S,12aR,14bR)-10- ( ( ( 2R,3R,4S, 5R, 6S )-6-( (benzyloxy )carbonyl)-3,4,5-tris( ( triethylsilyl )oxy )tetrahydro-2H-pyran-2-yl )oxy )-9-formyl-2,2, 6a, 6b, 9, 12a-hexamethyl-5-( ( triethylsilyl )oxy)-l,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,12b,13,14b-icosahydropicene-4a-carbonyl)oxy)-l-(prop-2-yn-l-yloxy)ethyl) dodecanedioate (46): To a stirred solution of 44 in DMF (100 mL) was added CS2CO3 (1.2 equiv.) and allowed to stir at 85 °C for 15 min. 45 (1.1 equiv.) was then added to it and allowed to stir at this temperature for next 1 hour until the complete conversion of 44.The reaction mixture was then allowed to cool down to room temperature, filtered, poured with ice cooled water and extracted with EtOAc (250 mL). The organic layer was washed with brine (100 mL), dried over Na2SC>4 and concentrated under reduced pressure. The concentrated crude was then purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 6 to 1 / 2) to afford 46 (55%) as white solid: Rf = 0.4 (EtOAc / Hep=l / l).

[0272] Synthesis of (2S,3S,4S,5R,6R)-6-(((3S,4S,4aR,6aR,6bS,8R,8aR,12aR,14bR)-8a-(((S)-2-((11 -carboxy undecanoy I )oxy )-2-(prop-2-yn-l -yloxy) ethoxy) carbonyl )-4-fonnyl-8-hydroxy-4,6a,6b,ll,ll,14b-hexamethyl-l,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (47): To a stirred solution of 46 in THF (100 mL) was added IM TBAF (0.5 equiv.) and allowed to stir for 6 hours until the complete deprotection of silyl group. The reaction mixture was concentrated under reduced pressure and dried in high vacuum for 3 hours. The crude concentrate was then dissolved in THF (50 mL) and Pd / C (10%) was added under H2 atmosphere (balloon) at room temperature. Upon completion of the reaction after 4 hours, the reaction mixture was filtered, concentrated53INIM-44840.601under reduced pressure, and purified by column chromatography (silica gel, MeOH / DCM = 1 / 10 to 1 / 4) to afford debenzylated intermediate 47 (65%) as a colorless syrup: Rf = 0.4 (MeOH / DCM =1 / 5 ).

[0273] Synthesis of (2S,3S,4S,5R,6R)-6-(((3S,4S,4aR,6aR,6bS,8R,8aR,12aR,14bR)-8a-(((S)-3-((ll-carboxyundecanoyl)oxy)-2-((l-(2-(((2R,3R,4S,5R,6R)-3-(((2S,3R,4S,5R,6S)-3,4-dihydroxy-6-methyl-5-( ((2S, 3R,4S, 5R )-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl )oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,5-dihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)ethyl)-lH-l,2,3-triazol-4-yl )methoxy )propoxy)carbonyl )-4-fonnyl-8-hydroxy-4, 6a, 6b, ll,ll,14b-hexamethyl-l,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,14,14a,14b-icosahydropicen-3-yl)oxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid (INI-5029): To a stirred solution of 16 or 19 and 47 (1.1 equiv.) in 100 mL of dioxane / water (5:1) was added sodium ascorbate (0.3 equiv.) and CuSO4 (0.15 equiv.) and allowed to stir at room temperature for 12 hours. After complete consumption of 16 or 19, the reaction mixture was filtered, concentrated and purified by column chromatography (silica gel; Methanol / DCM = 1 / 10 to 1 / 3) to afford 48 (55%) as white solid. Rf = 0.3 (MeOH / DCM=3 / 10).

[0274] Synthesis of tert-butyl (R)-( 3-( ( tert-butyldimethylsilyl)oxy)-2-hydroxypropyl)carbaniate (50): To a stirred solution of 49 in DMF (100 mL) was added te -Butyl(chloro)dimethylsilane (1.2 equiv.) and Imidazole (1.2 equiv.) at room temperature. The reaction was allowed to stir for 12 hours and quenched by adding ice cooled water, extracted with EtOAC (250 mL), washed with brine (100 mL) and dried over NazSCU. The organic layer was concentrated under reduced pressure and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 1 / 5) to afford targeted compound (85%) as a white solid: Rf = 0.4 (EtOAc / Hep=l / 4).

[0275] Synthesis of tert-butyl (R)-(3-((tert-butyldiniethylsilyl)oxy)-2-(prop-2-yn-l-yloxy)propyl)carbamate (51): To a stirred solution of 50 in DMF (100 mL) at 0 °C was added NaH (1.2 equiv.) and allowed to stir for 30 minutes. Propargyl bromide (1.1 equiv.) was then added dropwise into the reaction mixture and allowed to stir for 3 hours at room temperature. The reaction was quenched by adding ice cooled water and extracted with EtOAc (200 mL), washed with brine (100 mL) and dried over Na2SO4. The organic layer was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 3 / 10) to afford 25 (50%) as a white solid: Rf = 0.5 (EtOAc / Hep=l / l).54INIM-44840.601

[0276] Synthesis of tert-butyl (R)-(3-bromo-2-(prop-2-yn-l-yloxy)propyl)carbamate (52): To a stirred solution of 51 in THF (100 mL) was added IM TBAF (0.5 equiv.) and allowed to stir for 6 hours until the complete deprotection of silyl group. The reaction mixture was concentrated under reduced pressure and dried in high vacuum for 3 hours. The crude concentrate was then dissolved in DCM (150 mL) and CBr4 (2.5 equiv.) and TPP (2 equiv.) were subsequently added to it. The reaction was stirred for Ihour until the formation of 52 as dictated by TLC (EtOAc / Hep=l / 15). The crude mixture was then concentrated under reduced pressure and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 4 to 1 / 2) to afford 52 (60%) as a colorless liquid: Rf= 0.4 (EtOAc / Hep=l / 15).

[0277] Synthesis of ( R)-3-( ( tert-butoxycarbonyl )amino)-2-(prop-2-yn- 1 -yloxy )propyl (4aR,5R,6aS,6bR,8aR,9S,10S,12aR,14bR)-9-formyl-5,10-dihydroxy-2,2,6a,6b,9,12a-hexamethyl-l,3,4,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate (53): To a stirred solution of Quillaic acid in DMF (50 mL) was added CS2CO3 (1.2 equiv.) and allowed to stir at 85 °C for 15 min. 52 (1.1 equiv.) was then added to it and allowed to stir at this temperature for next 1 hour until the complete conversion of 52. The reaction mixture was then allowed to cool down to room temperature, filtered, poured with ice cooled water and extracted with EtOAc (100 mL). The organic layer was washed with brine (100 mL), dried over Na2SO4 and concentrated under reduced pressure. The concentrated crude was then purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 3 to 1 / 2) to afford 53 (55%) as white solid: Rf = 0.5 (EtOAc / Hep=l / l).

[0278] Synthesis of 55: Compound 53 was dissolved in a premixed solution of DCM / TFA (4:1) and allowed to stir 12 hours for room temperature. The crude mixture was then concentrated under reduced pressure and dried under high vacuum for 2 hours and used for next step without further purification. To a stirred solution of dried crude intermediate in anhydrous DMF (50 mL) at 0 °C was added HATU (1.2 equiv.) and TEA (3 equiv.) and allowed to stir for 15 minutes. Compound 6 (1.1 equiv.) was then added to the reaction mixture portion wise and allowed to stir at room temperature for 2 hours. After the completion of the reaction, it was quenched by adding water, extracted with DCM (100 mL), washed with brine (100 mL) and dried over Na2SO4. The organic layer was then concentrated under reduced pressure and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 10 to 1 / 5) to afford 55 (55%) as a white solid: Rf = 0.6 (EtOAc / Hep=l / 2).55INIM-44840.601

[0279] Synth esis of 3-( 12-(benzyloxy)-12-oxododecanamido)-2-((l-(2-(((2R,3R,4S,5R,6R)-3- (((2S, 3R, 4S, 5R, 6S )-3,4-dihydroxy-6-methyl-5-( ((2S, 3R, 4S, 5R)-3, 4, 5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,5-dihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)ethyl)-lH-l,2,3-triazol-4-yl)methoxy)propyl (4aR,5R,6aS,6bR,8aR,9S,10S,12aR,14bR)-9-formyl-5, 10-dihydroxy-2, 2, 6a, 6b, 9,12a-hexamethyl- l,3,4,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,12b,13,14b-octadecahydropicene-4a(2H)-carboxylate (56): To a stirred solution of Compound 55 (1 equiv.) and 16 or 19 (1.1 equiv.) in 100 mL of dioxane / water (5:1) was added sodium ascorbate (0.3 equiv.) and CuSO4 (0.15 equiv.) and allowed to stir at room temperature for 24 hours. After complete consumption of 16 or 19, the reaction mixture was filtered, concentrated and purified by column chromatography (silica gel; Methanol / DCM = 1 / 20 to 1 / 10) to afford 9 (55%) as white solid. Rf= 0.4 (MeOH / DCM=3 / 10).

[0280] Synthesis of 12-((2-((l-(2-(((2R,3R,4S,5R,6R )-3-(((2S,3R,4S,5R,6S)-3,4-dihydroxy-6- methyl-5-(((2S,3R,4S,5R)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,5-dihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)ethyl)-lH-l,2,3-triazol-4-yl )methoxy)-3-( ( ( 4aR, 5R, 6aS,6bR, 8aR, 9S, 1 OS, 12aR,14bR )-9-formyl-5, 1 O-dihydroxy-2,2,6a,6b,9,12a-hexamethyl-l,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,12b,13,14b- icosahydropicene-4a-carbonyl)oxy)propyl)amino)-12-oxododecanoic acid (INI-5034): To a stirred solution of 56 in THF (50 mL) was added Pd / C (10%) under H2 atmosphere (balloon) at room temperature. Upon completion of the reaction after 12 hours, the reaction mixture was filtered, concentrated under reduced pressure, and purified by column chromatography (silica gel, MeOH / DCM = 1 / 10 to 1 / 5) to afford the final product INI-5034 (65%) as a white powder: Rf = 0.5 (MeOH / DCM =1 / 4 ).

[0281] Compound (S)-2-((l-(2-(((2R,3R,4S,5R,6R)-3-(((2S,3R,4S,5R,6S)-3,4-dihydroxy-6- methyl-5-(((2S,3R,4S,5R)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran- 2-yl)oxy)-4,5-dihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)ethyl)- 1 H- 1,2,3-triazol-4- yl)methoxy)-3-(dodecanoyloxy)propyl (4aR,5R,6aS,6bR,8aR,9S,10S,12aR,14bR)-9-formyL 5,10-dihydroxy-2,2,6a,6b,9,12a-hexamethyl-l,3,4,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,12b,13,14b- octadecahydropicene-4a(2H)-carboxylate (INI-5036; 62 (55%; white as solid)) and compound INI-5037 (65%; white as solid); INI- 5032 (61%; white as solid) were prepared by the methods method above.56INIM-44840.601

[0282] Compounds 77-80 were synthesized using the methods described above. Compound 77 was purified by column chromatography (silica gel; EtOAc / hexanes = 1:4 to 1:2) to afford 77 (95%) as a colorless liquid (Rf = 0.4, EtOAc / hexanes = 1:2). Compound 78 was purified by column chromatography (silica gel; EtOAc / hexanes = 1:4 to 1:3) to afford 78 (60% over two steps) as a colorless liquid (Rf = 0.4, EtOAc / hexanes = 1:2). Compound 79 was purified by column chromatography (silica gel; EtOAc / hexanes = 1:10 to 1:5) to afford 79 (55%) as a white solid (0-anomer) (Rf = 0.7, EtOAc / hexanes = 1:3). Compound 80 was purified by column chromatography (silica gel; EtOAc / hexanes = 2:10 to 5:10) to afford 80 (75%) as a white solid (Rf = 0.3, EtOAc / hexanes = 1:1).

[0283] Synthesis of (3aS,4R, 6R, 7R, 7aR )-6-( 2-azidoethoxy)-4-( ( ( tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydro-4H-[l,3]dioxolo[4,5-c]pyran-7-ol (81):Using the general procedure of deacetylation as mentioned above, the crude mixture was filtered, concentrated and dried over high vacuum for 5 hours and used for next step without further purification to afford compound (60% crude) as a white solid. The crude was then dissolved in DMF (100 mL), TBSC1 (1.2 equiv.) and Imidazole (1.2 equiv.) were added at room temperature. The reaction was allowed to stir for 12 hours and quenched by adding ice cooled water, extracted with EtOAC (250 mL), washed with brine (100 mL) and dried over Na2SO4. The organic layer was concentrated under reduced pressure to afford compound 7 (85% crude) as a white solid (Lehtila, Reko L., et al. Tetrahedron, vol. 60, no. 16). The white solid was suspended in 100 mL of DCM / Acetone (3 / 1), and 2,2 dimethoxy propane (20 eq.) and / 2-Toluenesulfonic acid (0.1 eq.) was added subsequently. After stirring for 5 hours at room temperature, the reaction was quenched by adding sat. NaHCCh and extracted with DCM (200 mL), washed with brine (100 mL) and dried over NaiSOi. The crude mixture was concentrated and purified by column chromatography (silica gel; EtOAc / hexanes = 1 / 4 to 1 / 2) to afford 81 (55%) as an off-white solid: Rf = 0.6 (EtOAc / Hep=l / 3).

[0284] Synthesis of 82: (purified by column chromatography (silica gel; EtOAc / hep = 1 / 10 to 1 / 5) to afford 82 (50%) as a white solid (0-anomer): Rf = 0.6 (EtOAc / Hep=l / 3)) and 83 (purified by column chromatography (silica gel; MeOH / DCM = 1 / 100 to 1 / 10) to afford 83 (65% for 2 steps) as a white solid: Rf = 0.6 (MeOH / DCM=l / 10)) was completed by the above mentioned methods.

[0285] Synthesis of 1-benzyl 12-((2S)-2-((l-(2-(((2R,3R,4S,5R,6R)-3-(((2S,3R,4S,5R,6S)-3,4-dihydroxy-6-methyl-5-(((2S,3R,4S,5R)-3,4,5-trihydroxytetrahydro-2H-pyran-2-57INIM-44840.601yl )oxy )tetrahydro-2 H-pyran-2 -yl )oxy )-4, 5 -dihydroxy-6-(hydroxymethyl)tetrahydro-213-pyran-2 -yl)oxy)ethyl)-lH-l,2,3-triazol-4-yl)methoxy)-3-(((4aR,5R,6aS,6bR,8aR,9S,10S,12aR,14bR)-9-formyl-5, 10-dihydroxy-2,2, 6a, 6b, 9,12a-hexamethyl- l,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,ll,12,12a,12b,13,14b-icosahydropicene-4a- carbonyl)oxy)propyl) dodecanedioate (84): To a stirred solution of 83 and 74 (1.1 eq.) in 100 mL of dioxane / water (5:1) was added sodium ascorbate (0.3 eq.) and CuSO4 (0.15 eq.) and allowed to stir at room temperature for 24 hours. After complete consumption of 83, the reaction mixture was filtered, concentrated and purified by column chromatography (silica gel; Methanol / DCM = 1 / 20 to 1 / 3) to afford 84 (55%) as white solid. Rf = 0.4 (MeOH / DCM=l / 3).

[0286] Synthesis of 12-((2S)-2-((l-(2-(((2R,3R,4S,5R,6R)-3-(((2S,3R,4S,5R,6S)-3,4-dihydroxy- 6-methyl-5-(((2S,3R,4S,5R)-3,4,5-trihydroxytetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H- pyran-2-yl)oxy)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)ethyl)-lH- 1,2,3-triazol-4-yl)methoxy)-3-(((4aR,5R,6aS,6bR,8aR,9S, 1 OS, 12aR, 14bR)-9-formyl-5, 10- dihydroxy-2,2,6a,6b,9,12a-hexamethyl-l,2,3,4,4a,5,6,6a.6b,7,8,8a,9,10,11.12,12a,12b,13.14b- icosahydropicene-4a-carbonyl)oxy)propoxy)-12-oxododecanoic acid (INI-5030): To a stirred solution of 84 in THF (50 mL) was added Pd / C (10%) under H2 atmosphere (balloon) at room temperature. Upon completion of the reaction after 6 hours, the reaction mixture was filtered, concentrated under reduced pressure, and purified by column chromatography (silica gel, MeOH / DCM = 1 / 10 to 1 / 5) to afford the final product 85 (65%) as a white powder: Rf = 0.3 (MeOH / DCM =1 / 4 ).

[0287] Salting of the Compounds

[0288] The unsalted INI compound was weighed into a vial. 1,4-Dioxane and water are added to facilitate dissolving of the compound. Choline bicarbonate or TEA (2 equiv.) is added to the vial and the mixture is vortexed to complete the dissolution of the compound. If the compound is not dissolved after vortex then more 1,4-Dioxane and water is added until it is dissolved and the solution is clear. The vial contents are frozen at -80 °C and then lyophilized until dry.EXAMPLE 2

[0289] Cytokine Activity’

[0290] Compounds of formula I in aqueous formulation, alone and in combination with TLR4 agonist INI-2002 in liposomal formulation, were administered in vivo to mice and changes in58INIM-44840.601activity of cytokines / chemokines (IFN-y, TL-12p70, TNF-a, TNF-0, TP- 10, MCP-1, MIP-1, TNF-a, IL-ip, and IL-6) were measured in comparison to changes induced by QS-21 alone and in combination with INI-2002.

[0291] As shown in Tables 2 and 3, compounds were administered by i.v. An initial blood draw was taken 4 hours post-injection and a final blood draw 24 hours post-injection.

[0292] Referring now to FIG. 1 to FIG. 10. initial results demonstrated that representative compounds of formula (I) are able to modulate a subject’s immune system as shown by the increase of cytokine and chemokine activity.

[0293] Table 2. Novel Semi-Synthetic Saponin in vivo study designCompound Dose 1.15 1.3Qh 1.4ih;2H 2.5hr 3hr ' *4 4.5 Shr IS. Sh 6hr h * i hr hr hr I Q821 {Qvant; S 1 1 1 1 1 1 1 1 1 2 1 1 2002 liposome S 1 2 2 iill ijiioiiiiiiii:: o 1N1502S AO 50 1 1 2 1 1 1 2 1 1 2 1 1 iN!3026A« S||Sg|||S / 5f)|SSg / St 1 2 2 2 2 2 2 2 2 2 1 lNl5027Aq SO 1 1 1 1 2 1 1IBOI ioi ioisii<i’i 5825Aq / 2000po 50 / 5 1 I 2(2 / 3) 1 1 1 2 2 2 2 1 1 5026 Aq / 2002 lipo 50 / 5 1 1 2 2 2 2 2 2 5027 Aq / 2002 lipa 50 / 5 1 I I 1 2 1 2 2 2 2. 1 1 5028 Aq / 2002 lipo / 50 / 5 2 2 2 2 2 1 1 filliOioflOisi <2521 / 2032 lipa 5 / 5 1 1 212 / 3) 2 2 2 2 2 2 2 1 1t Blood collected at4 hours per orders As per AUP 012-22 Day of Dosing Gross Observation page 1 of 1 Noted Grimace, ruffling59INIM-44840.601

[0294] Table 3. Novel Semi-Synthetic Saponin in vivo study designCompound Dose; Route N i CageQS21 (Qvant) 5 IV 3 1 2. Q02 Liposome 5 IV 3 2 INI5025 Aq 50 IV 3 3 INI5Q26 Aq 50 IV 3 4 INI5027 Aq 50 IV 3 5 INI5028 Aq 50 IV 3 6 S025Aq / 2002 lipo 50 / 5 IV 3 7 5026 Aq / 2002 lipo 50 / 5 IV 3 8 5027 Aq / 2082 Lipo 50 / 5 IV 3 9 5028 Aq / 2002. Upo 50 / 5 IV 3 10QS21 / 2Q02 Upo 5 / 5 IV 3 11

[0295] Reactogenicity of the compounds also was measured. Results compiled in Tables 2 and 3 provided immediately hereinabove suggest that at the doses given, the compounds were reasonably-well tolerated.EXAMPLE 3Representative Data

[0296] Evaluation of the immunogenicity of salted formulations of INI-5026 and its analog, INI-5034.

[0297] This study demonstrates the immunogenicity of INI-5026 (salted and unsalted) as well as an analog, INI-5034. Mice (n=8) were immunized and boosted intramuscularly 14 days apart with each formulation (Table 4). Blood was collected 14 days post-primary injection (14dp 1) and again 14 days post-secondary injection (14dp2) for analysis of N2-specific serum antibody titers. At 14dp2, spleens were harvested for MSD cytokine analysis. Spleens were mechanically disaggregated, RBC lysed, and plated at 5 x 106cells / well. Splenocytes were stimulated with N2 (20 pg / mL) for 72 h, after which supernatants were collected to quantify secreted T cell-associated cytokines by MSD assay.60INIM-44840.601

[0298] Table 4. Salted formulation of novel saponins study design.! or w Ia1 | 32 j 1 3 a [ 1 34 ) 1 34 I S3 JHI-5325 / 110’2303 | 5 / 17 Owlme tok | §a Choice I IHI «23O | 5 / 13 JSl-5034 | 5 3| 5 / 1 511 | 5 a12 | 5 / 1 313 TBA 1 514 Wb502BT£A / IH^2W2 | 5 / 1

[0299] Analysis of N2-specific titers revealed that INI-5026 (salted an unsalted) and 5034 (unsalted) induced, on average, increased IgG2a (Thl-biased) responses compared to QS ■>- -21 alone, with INI-5034 (choline bicarb.) reaching statistical significance (FIG. 11 ). In contrast, INI-5026 (choline bicarb.) resulted in significantly higher IgGl (Th2-biased) responses, while all other monotherapies induced comparable or slightly decreased levels of IgGl.

[0300] All T cell- associated cytokines were similar between QS-21 and each monotherapy, with the exception of TNF-a and IL-5 (FIG. 12). Notably, the expression of IFN-y, IL-17A, and TNF-a for formulations of INI-5026 (salted and unsalted) were increased compared to QS-21 alone. Importantly, the monotherapy formulations did not demonstrate synergistic activity when combined with INI-2002 regarding humoral or cell-mediated responses.

[0301] Evaluation of the immunogenicity of INI-5026 and INI-5028.

[0302] The impact of 1:1 and 5:1 dose ratios of the novel semi-synthetic saponins, INI-5026 and INI-5028, and the TLR4 agonist, INI-2002, on synergy was assessed. Female BALB / c mice (n=7 / group) were immunized and boosted intramuscularly 14 days apart with each formulation61INIM-44840.601(Table 5). Blood was collected at day 14 post-secondary injection (14dp2) for analysis of N2-specific serum antibody titers. At 14dp2, spleens were harvested, mechanically disaggregated, RBC-lysed, and plated at 5 x 106cells / well in a 96-well plate. Splenocytes were then stimulated with the influenza antigen, N2 (20 pg / mL), for 72 h, after which supernatants were collected to quantify secreted T cell-associated cytokines.

[0303] Table 5. Novel saponins study design.A / Vic N2 INI-2002 QS-21 Novel SapGroup Compound M M M W (Pg) n1 iliiJJJJJJ 0.3 |||||||| 6 2 N2 1 1 1 6 3 IlJIlJJJIJJIJjJJ |Jj «* Jill lllilll >• 7 4 INI-2002 liposome 1 1 7 III j| '•C III i ll III II III 76 INI-5026 1 5 7 7 ■liiijjjjii IJJIIJ^ lllll 5 7 8 INI-5026 / INI-2002 1 1 5 7 9 INI-5028 / INI-2002 jiijj liiiliiii 5 7 19 INI-5026 / INi-2002 * t 5 5 7 11 INI-5028 / INI-2002 -lllll 5 7

[0304] Analysis of N2-specific serum antibody titers revealed that, while INI-5026 and INI-5028 alone induced IgG2a (Th 1 -biased) and IgGl (Th2-biased) responses comparable to that induced by QS-21 alone, combination with INI-2002 at either dose ratio did not result in observable synergy (FIG. 13). All T cell-associated cytokines were similarly comparable between INI-5026, INI-5028, and QS-21 (FIG. 14).

[0305] Although the foregoing subject matter has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be understood by those skilled in the art that certain changes and modifications can be practiced within the scope of the appended claims.62INIM-44840.601

Claims

WHAT IS CLAIMED IS:

1. A compound of formula (la) or formula (lb):wherein:the compound of formula (la) or formula (lb) can be in the (R) configuration, the (S) configuration, or a mixture of (R, S) configurations;Ri is selected from H, a monosaccharide, a disaccharide, and a moiety selected from:Reach n is independently an integer selected from 0, 1, 2, 3, 4, 5, 6, 7. 8, 9.

10.

11. 12, 13, 14, and 15;R is selected from an aromatic, a saturated or unsaturated, branched or straight chain alkyl; R2 is selected from H, a carboxylic acid, an amide, phenyl, halogen, and an aromatic; W is O or N;X is O, N, or S;Y is selected from -C(O)(CH2)nCOOH, -(O)(CH2)CONH2, Ar, biaryl, branched alkyl chain, saturated alkyl chain, unsaturated alkyl chain, and halogen;Z is selected from D-Fucose, L-Fucose, and galactose; andpharmaceutically acceptable salts thereof.

2. The compound of claim 1, wherein the compound is a compound of formula (la):63INIM-44840.601(R), (S), (R, S)(la);wherein:Ri is selected from a monosaccharide, a disaccharide, H, and a moiety selected from:RR is selected from an aromatic, a saturated, or unsaturated, branched or straight alkyl chain; R2 is selected from a carboxylic acid, an amide, phenyl, halogen, an aromatic, and H; each n is independently an integer selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, and 15;W is O or N;X is O, N, or S;Y is selected from -C(O)O(CH2)nCOOH, -(O)(CH2)CONH2. Ar, biaryl, branched alkyl chain, saturated alkyl chain, unsaturated alkyl chain, and halogen;Z is selected from D-Fucose, L-Fucose, and galactose; andpharmaceutically acceptable salts thereof.

3. The compound of claim 1, wherein the compound is a compound of formula (lb):64INIM-44840.6014. The compound of claim 1, wherein the compound of formula (la) is:wherein: R is selected from -OCH3, -OH, -NH2, -O(CH2)nCOOH, and -NH(CH2)nCOOH.

5. The compound of claim 1, wherein the compound of formula (la) is selected from:65INIM-44840.601INIM-44840.6016. The compound of claim 1, wherein the compound of formula (Ia) is selected from:67INIM-44840.601INIM-44840.6017. A method for modulating an immune system of subject, the method comprising administering a compound of any one of claims 1 to 6 to the subject.

8. A method for treating or preventing a disease, disorder, or condition, the method comprising administering an effective amount of a compound of any one of claims 1 to 6 to a subject in need of treatment thereof.

9. The method of claim 8, wherein the disease, disorder, or condition is selected from cancer, a bacterial infection, a viral infection, a fungal infection, a parasitic infection, an immune-mediated disorder, a central nervous system disease, a peripheral nervous system disease, a neurodegenerative disease, a mood disorder, a sleep disorder, a cerebrovascular disease, a peripheral artery disease, and a cardiovascular disease.

10. The method of claim 9, wherein the disease, disorder, or condition is cancer.

11. The method of claim 10, wherein the cancer is selected from colorectal cancer, aero-digestive squamous cancer, lung cancer, brain cancer, liver cancer, stomach cancer, sarcoma,69INIM-44840.601leukemia, lymphoma, multiple myeloma, ovarian cancer, uterine cancer, breast cancer, melanoma, prostate cancer, bladder cancer, pancreatic carcinoma, and renal carcinoma.

12. The method of claim 9, wherein the disease, disorder, or condition is an infection.

13. The method of claim 12, wherein the infection comprises a bacterial, fungal, protozoal or viral infection.

14. The method of claim 13, wherein the bacterial, fungal, protozoal, or viral infection is selected from tuberculosis and mycobacterium avium, leprosy; pneumocystis carnii, cryptosporidiosis, histoplasmosis, toxoplasmosis, trypanosome infection, leishmaniasis, and infections caused by bacteria of the genus Escherichia, Enterobacter, Salmonella, Staphylococcus, Klebsiella, Proteus, Pseudomonas, Streptococcus, and Chlamydia, and fungal infections, including candidiasis, aspergillosis, histoplasmosis, cryptococcal meningitis, a viral disease, including respiratory syncytial virus (RSV), hepatitis B, hepatitis C, Dengue virus, herpes simplex virus, including HSV-I, HSV-II, CMV, and VZV, molluscum contagiosum, vaccinia, variola, lentivirus, human immunodeficiency virus (HIV), human papilloma virus (HPV), cytomegalovirus (CMV), varicella zoster virus (VZV), rhinovirus, enterovirus, adenovirus, coronavirus, including SARS, influenza, para-influenza, mumps virus, measles virus, papovavirus, hepadnavirus. flavivirus, retrovirus, arenavirus, including LCM, Junin virus, Machupo virus, Guanarito virus and Lassa Fever, and filovirus, including Ebola virus and Marburg virus.

15. The method of any one of claims 8 to 14, further comprising administering a therapeutic agent.

16. The method of claim 15, wherein the therapeutic agent is selected from an immunomodulating agent, antiviral agent, an anti-inflammation agent, a chemotherapeutic agent, an anti-cancer vaccine, and hormonal therapy.

17. The method of claim 16, wherein the immunomodulating agent comprises a vaccine adjuvant.70INIM-44840.60118. The method of claim 16, wherein the immunomodulating agent is selected from a liposome, alum, Freund's complete or incomplete adjuvant, a detoxified endotoxin, and a Toll-Like Receptor agonist.

19. The method of claim 18. wherein the Toll-like Receptor agonist is selected from TLR2, TLR3, TLR2 / 3, TLR4, TLR5, TLR7, TLR8, TLR7 / 8, and a TLR 9 agonist.

20. The method of claim 19, wherein Toll-like Receptor agonist comprises a TLR4 agonist.

21. The method of claim 20, wherein the TLR4 agonist is selected from natural and synthetic Lipid A derivatives and mimetics.

22. The method of claim 21, wherein the natural and synthetic Lipid A derivatives and mimetics are selected from monophosphoryl lipid A, MPL, MPLA, 3D-MPL, GLA INI-2002, and INI-2004.

23. The method of any one of claims 8 to 14, further comprising administering an antigen.

24. The method of claim 23, wherein the antigen is an antigen associated with Varicella zoster virus (VZV).

25. The method of claim 23, wherein the antigen comprises an allergen.

26. The method of claim 25, wherein the allergen is a respiratory allergen or a food allergen.

27. The method of claim 26, wherein the respiratory antigen is selected from pollen, dander, and dust.71INIM-44840.60128. The method of claim 26, wherein the food allergen comprises a nut allergen.72INIM-44840.601

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