Method and compound for regulating immune response

Abstract

The present disclosure provides a method for treating inflammatory conditions by administering a therapeutically effective amount of a small molecule compound that modulates Rac2 activity by interfering with the binding between Rac2 and RhoGDI to a subject requiring treatment. The small molecule compound reduces the expression of thymic stromal lymphocyte-stimulating factor (TSLP). The present method may be used to treat atopic dermatitis or asthma. The small molecule compound can bind to residues E171 and / or L155 of Rac2, has an EC50 for binding to Rac2 of less than 10 pM, and exhibits at least three times greater binding affinity for Rac2 compared to Rae1 and Rac3.

Description

Technology Field

[0001] Cross-reference regarding related applications

[0002] The present application claims priority under 35 USC § 119(e) to U.S. provisional application USSN 63 / 622,064 filed January 18, 2024, and U.S. provisional application USSN 63 / 745,795 filed January 16, 2025, the full text of each of which is incorporated herein.

[0003] Technology field

[0004] The present disclosure relates to a method and a compound for modulating an immune response, and more particularly to a small molecule compound that selectively targets Rac2 to reduce thymic stromal lymphocyte-stimulating factor (TSLP) expression for the treatment of inflammatory pathologies. Background Technology

[0005] Inflammatory conditions such as atopic dermatitis and asthma affect millions of people worldwide, resulting in significant morbidity and reduced quality of life. These conditions are characterized by dysregulation of immune responses involving various cytokines and signaling pathways. Thymic stromal lymphocyte-stimulating factor (TSLP) has emerged as a key mediator in the pathogenesis of allergic inflammatory disorders.

[0006] TSLP is an epithelial cell-derived cytokine that plays a crucial role in initiating and maintaining type 2 inflammatory responses. It acts on various cell types, including dendritic cells, mast cells, and T cells, to promote allergic inflammation. Elevated levels of TSLP have been observed in skin lesions of patients with atopic dermatitis and in the airways of asthmatic individuals.

[0007] Current treatments for inflammatory conditions such as atopic dermatitis and asthma include topical corticosteroids, systemic immunosuppressants, and biological therapies targeting specific cytokines or their receptors. While these approaches can be effective, they may be associated with side effects, particularly with long-term use. Additionally, some patients do not respond adequately to existing therapies, highlighting the need for new treatment strategies.

[0008] Recent research has focused on understanding the molecular mechanisms regulating TSLP expression and identifying potential targets for therapeutic intervention. Small GTPases of the Rho family, including Rac proteins, are involved in various cellular processes and signaling pathways related to inflammation. However, the specific roles of individual Rac proteins in regulating TSLP expression and inflammatory responses have not yet been fully elucidated.

[0009] Developing targeted approaches to modulate key signaling pathways involved in TSLP production and inflammatory responses could potentially provide new therapeutic options for patients with allergic inflammatory disorders. These strategies could offer improved efficacy and safety profiles compared to existing treatments.

[0010] Brief summary

[0011] This summary is provided to introduce a selection of concepts in a simplified form, which is further detailed in the detailed description below. This summary is not intended to identify the core or essential characteristics of the claimed subject matter, nor is it intended to be used to assist in determining the scope of the claimed subject matter.

[0012] According to one aspect of the present disclosure, a method for treating an inflammatory pathological condition is provided. The method comprises administering a therapeutically effective amount of a small molecule compound that modulates Rac2 activity by interfering with the binding between Rac2 and RhoGDI to a subject requiring treatment. The small molecule compound reduces the expression of thymic stromal lymphocyte-stimulating factor (TSLP).

[0013] According to another aspect of the present disclosure, the method may include one or more of the following characteristics. The inflammatory condition may be atopic dermatitis. Administration of the small molecule compound may result in a reduction in the dermatitis severity score. The inflammatory condition may be asthma. The small molecule compound may reduce TSLP expression by at least 50% compared to a placebo. The small molecule compound may bind to at least one of residues E171 and L155 of Rac2. The small molecule compound may have an EC50 for binding to Rac2 of less than 10 μM. The small molecule compound may exhibit at least three times greater binding affinity for Rac2 compared to Rac1 and Rac3.

[0014] According to another aspect of the present disclosure, a method for reducing thymic stromal lymphocyte-stimulating factor (TSLP) expression in a subject is provided. The method comprises administering to a subject a therapeutically effective amount of a small molecule compound that exhibits increased binding affinity to Rac2 compared to Rac1 and Rac3. The small molecule compound interferes with the interaction between Rac2 and RhoGDI.

[0015] According to another aspect of the present invention, the method may include one or more of the following characteristics. The small molecule compound may exhibit a binding affinity for Rac2 that is at least three times greater than that for Rac1 and Rac3. Administration of the small molecule compound may result in at least a 50% reduction in TSLP production compared to a placebo. The small molecule compound may have an EC50 for binding to Rac2 of less than 10 μM. The small molecule compound may bind to at least one of residues E171 and L155 of Rac2.

[0016] According to another aspect of the present disclosure, a method for regulating Rac2 activity is provided. The method comprises administering a therapeutically effective amount of a small molecule compound that binds to Rac2 to a subject.

[0017] According to another aspect of the present disclosure, the method may include one or more of the following characteristics. The small molecule compound may interfere with the interaction between Rac2 and RhoGDI. The small molecule compound may bind to at least one of residues E171 and L155 of Rac2. The small molecule compound may have an EC50 for binding to Rac2 of less than 10 μM. Administration of the small molecule compound may result in at least a 50% reduction in thymic stromal lymphocyte-stimulating factor (TSLP) expression compared to a placebo. The small molecule compound may exhibit increased binding affinity for Rac2 compared to Rac1 and Rac3. The small molecule compound may exhibit at least three times greater binding affinity for Rac2 compared to Rac1 and Rac3.

[0018] The foregoing general description of exemplary embodiments and the following detailed description thereof are merely illustrative of the teachings of the present disclosure and are not limiting. Brief explanation of the drawing

[0019] Non-limiting and non-comprehensive examples are described with reference to the drawings below. FIGS. 1a, 1b, and 1c illustrate system diagrams showing the binding characteristics and structural characteristics of compound II and RAC2 protein according to an embodiment of the present disclosure. FIGS. 2a, 2b, and 2c depict the study design and immune cell analysis from an experiment investigating therapeutic effects in mice according to an embodiment. FIGS. 3a, 3b, and 3c illustrate the study design and results from an experiment investigating the therapeutic effect of compound II in mice according to an embodiment of the present disclosure. Figures 4a and 4b illustrate the effects of different treatments on serum IgE and eotaxin concentrations in an inflammation model according to an embodiment. FIGS. 5a and 5b illustrate the effects of different treatments on MCP-1 levels and peribronchial inflammation in an inflammation model according to an embodiment of the present disclosure. FIG. 6 illustrates a heatmap visualization comparing gene expression patterns across different experimental conditions related to atopic dermatitis according to an embodiment. FIGS. 7a and 7b illustrate a study design using dermatitis severity scores over time and mouse skin photographs according to an embodiment of the present disclosure. Figures 8a and 8b depict a schematic diagram of the study design and the measurement of dermatitis severity in a mouse model according to an embodiment. FIG. 9 illustrates a bar graph of drainage change in response to different treatments according to an embodiment of the present disclosure. FIGS. 10a and 10b illustrate histological images and quantitative analysis of epidermal thickness across different treatment conditions according to an embodiment. FIGS. 11a, 11b, and 11c depict box plots showing Rac2 expression levels across different experimental conditions and disease states according to an embodiment of the present disclosure. Figures 12a and 12b illustrate a cell heat migration assay of FLAG-Rac2 protein stability according to an embodiment. FIGS. 13a, 13b, 13c, and 13d illustrate volcano plots showing differential protein expression analysis in human and mouse cell lines according to an embodiment of the present disclosure. FIG. 14 depicts a bar graph showing the relative abundance of Rac2 binding partners after ENV-294 treatment according to an embodiment. FIGS. 15a, 15b, 15c, and 15d illustrate peptide sequence analysis results and dose-response curves for RAC proteins according to aspects of the present disclosure. In FIG. 15a, the sequences indicated correspond (from top to bottom) to SEQ ID NOs: 22 (AKWFPEVR); 23 (HHCPSTPIILVGTK); 24 (KLAPITYPQGLALAK); 25 (LAPITYPQGLALAK); 26 (WFPEVR); and 27 (LDLRDDKDTIEK). In FIG. 15b, the sequences indicated correspond (from top to bottom) to SEQ ID NOs: 28 (TVFDEAIR); 22 (AKWFPEVR); 23 (HHCPSTPIILVGTK); 24 (KLAPITYPQGLALAK); 27 (LDLRDDKDTIEK); and 29 (AKWYPEVR). FIGS. 16a, 16b, 16c, and 16d illustrate volcano plots showing differential protein expression analysis in human and mouse cell lines according to an embodiment. FIGS. 17a and 17b depict cell type distribution in Th2 human explants and MC903 mouse skin samples treated with Compound II according to an aspect of the present disclosure. FIG. 18 illustrates the immunohistochemical staining of skin tissue sections under different experimental conditions according to an embodiment. FIGS. 19a, 19b, and 19c illustrate the effects of different treatments on serum IgE, tissue IL-31, and tissue IL-13 levels according to an embodiment of the present disclosure. FIGS. 20a and 20b depict box plots showing tissue levels of TSLP and Krt6a across different treatment groups according to an embodiment. FIGS. 21a, 21b, 21c, and 21d illustrate cell expression and functional characteristics across different cell types and tissues according to an embodiment of the present disclosure. FIG. 22 illustrates a system diagram showing the mechanism of action of compound II in regulating TSLP production in atopic dermatitis according to an embodiment. FIGS. 23a, 23b, 23c, 23d, 23e, 23f, and 23g depict box plots showing gene expression levels for JAK-STAT pathway components across therapeutic conditions according to an embodiment of the present disclosure. FIGS. 24a and 24b show volcano plots of differential gene expression analysis comparing Th2 stimulation with a vehicle control and compound II treatment according to an embodiment. Specific details for implementing the invention

[0020] The present disclosure relates to a method for treating inflammatory conditions by modulating the immune response through targeted intervention of intracellular signaling pathways. In particular, the method described herein includes the use of a small molecule compound that selectively modulates the activity of Rac2, a member of the Rho family of GTPases involved in various cellular processes including immune cell function and inflammatory response.

[0021] The small molecule compound described herein may act by interfering with the interaction between Rac2 and its regulatory protein, thereby causing alterations in the downstream signaling cascade. This regulation of Rac2 activity may result in a decrease in the expression of thymic stromal lymphocyte-stimulating factor (TSLP), a key cytokine associated with various inflammatory conditions.

[0022] In some cases, the method described herein may be particularly effective in treating atopic dermatitis, a chronic inflammatory skin condition characterized by severe itching and eczematous lesions. Modulation of Rac2 activity and subsequent reduction of TSLP expression may help alleviate symptoms and underlying inflammation associated with atopic dermatitis.

[0023] The method disclosed herein may also be applicable to other inflammatory conditions such as asthma. By targeting Rac2 and regulating TSLP expression, the small molecule compound may help reduce airway inflammation and improve respiratory function in individuals suffering from asthma.

[0024] The selective nature of this small molecule compound in targeting Rac2 may provide advantages over broader immunosuppressive therapies, potentially offering a more targeted approach to treating inflammatory conditions while reducing the risk of systemic side effects. The method described herein may represent a novel therapeutic strategy for managing chronic inflammatory diseases through the modulation of specific intracellular signaling pathways.

[0025] This small molecule compound is a novel therapeutic agent designed to regulate Rac2 activity and reduce thymic stromal lymphocyte-stimulating factor (TSLP) expression. The compound's chemical structure and physical properties contribute to its unique binding characteristics and selectivity for Rac2 compared to related proteins.

[0026] FIGS. 1a, 1b, and 1c illustrate the binding characteristics and structural properties of the small molecule compound with respect to the Rac2 protein. The compound may have a molecular weight in the range of about 250 Da to about 500 Da, alternatively about 300 Da to about 450 Da, and alternatively about 375 Da. In some cases, the compound may exist in various salt forms including, but not limited to, hydrochloride, sulfate, or sodium salts.

[0027] This small molecule compound demonstrates high affinity and selectivity for Rac2. As illustrated in Fig. 1a, the compound exhibits dose-dependent competition with the photoactivated probe for binding to FLAG-Rac2. The apparent EC50 for binding to Rac2 may range from about 1 μM to about 15 μM, alternatively from about 3 μM to about 10 μM, and alternatively about 7 μM. This high affinity binding is further supported by the data presented in Figs. 13a, 13b, 13c, and 13d, which show significant changes in Rac2 protein levels after treatment with this small molecule compound in both human dHL60 and mouse RAW264.7 cell lines.

[0028] The selectivity of this small molecule compound for Rac2 relative to related proteins is a key characteristic of its mechanism of action. The compound exhibits a greater binding preference for Rac2 compared to Rac1, with selectivity ratios ranging from about 3 to about 8, alternatively from about 4 to about 7, and alternatively from about 5.7. Similarly, the compound demonstrates selectivity for Rac2 relative to Rac3, with selectivity ratios ranging from about 2 to about 6, alternatively from about 3 to about 5, and alternatively from about 3.9.

[0029] Binding of the present small molecule compound to Rac2 can involve specific residues on the protein. As exemplified in FIGS. 1b and 1c, the compound can interact with at least one of residues E171 and L155 of Rac2. These residues are located outside the GTP binding domain and at the C-terminus of Rac2, which can contribute to the selectivity of the compound toward other Rac family members. In various embodiments, the amino acid sequence of Rac2 is MGSSHHHHHH (SEQ No. 1), SSGLVPRGSH (SEQ No. 2), MQAIKCVVVG (SEQ No. 3), DGAVGKTCLL (SEQ No. 4), ISYTTNAFPG (SEQ No. 5), EYIPTVFDNY (SEQ No. 6), SANVMVDSKP (SEQ No. 7), VNLGLWDTAG (SEQ No. 8), QEDYDRLRPL (SEQ No. 9), SYPQTDVFLI (SEQ No. 10), CFSLVSPASY (SEQ No. 11), ENVRAKWFPE (SEQ No. 12), VRHHCPSTPI (SEQ No. 13), ILVGTKLDLR (SEQ No. 14), DDKDTIEKLK (SEQ No. 15), EKKLAPITYP (Sequence No. 16), QGLALAKEID (Sequence No. 17), SVKYLECSAL (Sequence No. 18), TQRGLKTVFD (Sequence No. 19), EAIRAVLCPQ (Sequence No. 20), PTRQQKRAC (Sequence No. 21).

[0030] Cellular Thermal Shift Assay (CETSA) is a powerful technique used to evaluate the binding of small molecules to target proteins within a cellular environment. Figures 12a and 12b present CETSA data providing significant evidence regarding the interaction between this small molecule compound and Rac2. The compound demonstrates a significant thermal stabilizing effect on Rac2, inducing a thermal shift of more than 2°C upon binding. This shift, ranging from approximately 2°C to approximately 5°C with typical values ​​of 2.5°C to 4°C and an average of approximately 3°C, indicates a strong and specific interaction between the compound and Rac2.

[0031] Importantly, this heat transfer is observed only in cell lysates, not in purified Rac2 protein. This distinction suggests that the binding of the compound to Rac2 depends on specific cellular conditions or requires the formation of protein complexes present in the cellular environment. This context-dependent binding is an important characteristic of the compound's mechanism of action because it can contribute to its selectivity and efficacy in regulating Rac2 activity within physiologically relevant settings.

[0032] The magnitude of the heat shift provides quantitative information regarding the strength of the interaction between the compound and Rac2. A shift of 2–5°C is considered significant in CETSA experiments and typically indicates high-affinity binding events. This data confirms other binding assays, such as competitive binding studies and chemoprepetic analyses presented in the previous figure, thereby further validating Rac2 as the primary target of this small molecule compound.

[0033] The context-dependent nature of the binding holds significant implications for the therapeutic potential of the compound, as revealed by CETSA data. This suggests that the compound preferentially interacts with Rac2 in its natural cellular environment, potentially minimizing off-target effects and enhancing its specificity for regulating Rac2-dependent inflammatory pathways. This feature may contribute to the favorable efficacy and safety profiles of the compound observed in preclinical studies of inflammatory conditions such as atopic dermatitis and asthma.

[0034] The interaction between this small molecule compound and Rac2 results in a significant disruption of protein-protein interactions, particularly the binding between Rac2 and its regulatory protein RhoGDI (Rho GDP-dissociation inhibitor). This disruption is a key aspect of the compound's mechanism of action in regulating Rac2 activity. Figure 14 provides quantitative evidence of this effect, demonstrating that treatment with the compound leads to a substantial reduction in the formation of the Rac2-RhoGDI complex.

[0035] The magnitude of this reduction is noteworthy, with the decrease in the Rac2-RhoGDI complex ranging from approximately 5 to 12 times compared to the untreated state. More specifically, the data suggest that the reduction typically falls within a narrower range of about 6 to 10 times, indicating an average reduction of approximately 8 times. This significant reduction in complex formation demonstrates the compound's potent effect on the regulatory mechanism of Rac2.

[0036] Disruption of the Rac2-RhoGDI interaction has significant implications for Rac2 function and subsequent cellular processes. RhoGDI plays a crucial role in regulating Rho GTPases, including Rac2, by maintaining them in an inactive GDP-bound state within the cytoplasm. By disrupting this interaction, this small molecule compound can alter the normal cycle of Rac2 between its active (GTP-bound) and inactive (GDP-bound) states, as well as modify its intracellular localization.

[0037] This regulation of Rac2 activity through the disruption of RhoGDI and its interactions may contribute to downstream effects of the compound, including the decrease observed in thymic stromal lymphocyte-stimulating factor (TSLP) expression. Modified Rac2 activity can influence various signaling pathways and cellular processes involved in inflammatory responses, potentially leading to therapeutic effects observed in conditions such as atopic dermatitis and asthma.

[0038] The quantitative range of reduction in the Rac2-RhoGDI complex provides valuable information for understanding the efficacy of the compound and potentially optimizing administration strategies for future clinical applications. Furthermore, this data supports the specificity of the compound's action on Rac2, as the magnitude of the effect on the Rac2-RhoGDI interaction is substantial and consistent across multiple studies.

[0039] The chemoprequency analyses presented in Figures 16a, 16b, 16c, and 16d provide strong evidence of the high specificity of this small molecule compound for Rac2. This comprehensive analysis involves the systematic identification and quantification of proteins interacting with or affected by the compound across the whole proteome of both human and mouse cell lines. The generated volcano plots provide a visual representation of the statistical significance and magnitude of protein changes in response to compound treatment.

[0040] In these plots, each point represents a single protein, the x-axis typically shows a log2-fold change in protein abundance or interaction, and the y-axis represents the negative log10 of the p-value. This arrangement makes it easy to identify proteins that exhibit both large and statistically significant changes. The term "volcano" refers to the characteristic shape formed when many proteins show small or minor changes (clustered at the bottom), while a few proteins demonstrate large and significant changes (appearing at the top of the plot, resembling a volcanic eruption).

[0041] The fact that Rac2 stands out in these plots and exhibits statistically significant abundance scores in both human and mouse cell lines strongly supports the selectivity of the compound. This observation is particularly noteworthy considering that thousands of proteins were analyzed simultaneously. The consistency of these results across species (human and mouse) further validates the robustness of the compound's specificity for Rac2.

[0042] Furthermore, the use of both human and mouse cell lines in this analysis strengthens the clinical applicability of the compound. This suggests that specific interactions between the compound and Rac2 are conserved across species, which is often a critical factor in predicting the success of therapeutic approaches from preclinical models to human application.

[0043] The statistically significant abundance score for Rac2 indicates that the interaction between the compound and Rac2 is highly unlikely to be coincidental. This statistical rigor adds significant weight to the conclusion that Rac2 is indeed the primary target of this small molecule compound, further supporting its potential as a specific regulator of Rac2 activity in the treatment of inflammatory conditions such as atopic dermatitis and asthma.

[0044] In some cases, the small molecule compound may be modified to produce derivatives or analogs having similar or improved properties. These modifications may include, but are not limited to, the addition of functional groups to improve solubility, the incorporation of isotopes for tracking or imaging, or conjugation with targeting moiety for improved tissue-specific delivery. The molecular weight of such derivatives may be in the range of about 300 Da to about 1000 Da, alternatively about 400 Da to about 800 Da, and alternatively about 600 Da.

[0045] The small molecule compound may be formulated for various routes of administration, including but not limited to oral, topical, intravenous, subcutaneous, or inhalation. The compound may demonstrate stability in aqueous solutions having a pH in the range of about 4 to about 9, alternatively about 5 to about 8, and alternatively about 7. The solubility of the compound in an aqueous medium may be in the range of about 0.1 mg / mL to about 50 mg / mL, alternatively about 1 mg / mL to about 20 mg / mL, and alternatively about 10 mg / mL.

[0046] This small molecule compound regulates Rac2 activity through a specific mechanism that ultimately leads to a decrease in thymic stromal lymphocyte-stimulating factor (TSLP) expression. This regulation occurs primarily through the disruption of the binding between Rac2 and its regulatory protein, RhoGDI.

[0047] In some cases, this small molecule compound can cause an eightfold reduction in RhoGDI binding to Rac2. This disruption of the Rac2-RhoGDI interaction alters the membrane-cytoplasmic distribution of Rac2, which may affect its availability for downstream signaling. This altered distribution may play an important role in regulating the GTPase cycle of Rac2 without completely blocking its function.

[0048] The mechanism of action of this small molecule compound in regulating TSLP production in atopic dermatitis is a complex process involving various cell types and signaling pathways. As illustrated in Fig. 22, the compound exerts its effects by targeting Rac2, a small GTPase protein, in various cell types that play a significant role in the pathogenesis of atopic dermatitis. These cell types include Langerhans cells, which are specialized dendritic cells in the epidermis; dendritic cells, which are key antigen-presenting cells; basophils and mast cells, which are important effector cells in allergic reactions; and keratinocytes, which are the dominant cell type in the epidermis.

[0049] In these various cell types, the interaction between Rac2 and the compound leads to a series of events that ultimately result in reduced TSLP production. TSLP, or thymic stromal lymphocyte-stimulating factor, is a key cytokine in the initiation and maintenance of allergic inflammation in atopic dermatitis. By reducing TSLP levels, the compound effectively interrupts a key step in the disease-associated inflammatory cascade.

[0050] Regulation of Rac2 by this small molecule compound has the potential to affect various cellular processes in each of these cell types. For example, in Langerhans cells and dendritic cells, Rac2 inhibition can alter their migration, maturation, and ability to present antigens to T cells. In basophils and mast cells, this can affect degranulation and the release of inflammatory mediators. In keratinocytes, Rac2 regulation can affect barrier function and the production of pro-inflammatory cytokines.

[0051] This multicellular approach to reduce TSLP production represents a novel and potentially more comprehensive strategy for the management of atopic dermatitis. By targeting upstream regulators such as Rac2, the compound may offer advantages over therapies that target individual inflammatory mediators or specific cell types. The broad impact across multiple cell types involved in the disease process suggests that this approach can simultaneously address various aspects of the pathophysiology of atopic dermatitis, potentially leading to more effective disease control.

[0052] This small molecule compound can modulate Rac2 function without directly interfering with its underlying GTPase activity. This selective modulation allows for the regulation of specific downstream pathways while preserving essential cellular functions mediated by Rac2. In some cases, this may result in a more targeted therapeutic approach with potentially fewer side effects compared to broader GTPase inhibitors.

[0053] The effects of the present small molecule compound on the JAK-STAT pathway, a critical signaling cascade in inflammatory responses, are comprehensively illustrated in FIGS. 23a to 23g. These figures present a detailed analysis of changes in gene expression for key components of the pathway, including Janus kinases (Jak1, Jak2, Jak3), signaling and transcription activators (Stat1, Stat2, Stat3), as well as tyrosine kinase 2 (Tyk2). The JAK-STAT pathway is closely linked to TSLP signaling because TSLP receptor activation leads to phosphorylation of JAK proteins, which in turn activates STAT proteins, ultimately resulting in the transcription of pro-inflammatory genes.

[0054] Changes observed in the expression levels of these JAK-STAT pathway components across different therapeutic conditions provide valuable insights into the mechanisms by which these small molecule compounds regulate inflammatory responses. For example, changes in the expression of Jak1, Jak2, and Jak3 may affect the early stages of TSLP signaling, whereas changes in Stat1, Stat2, and Stat3 may affect downstream transcriptional responses. The inclusion of Tyk2 in this analysis is particularly relevant because it is known to play a role in IL-13 signaling, another key cytokine in the pathogenesis of atopic dermatitis.

[0055] The regulation of these JAK-STAT pathway components by this small molecule compound is likely to contribute significantly to the overall decrease in TSLP expression. This effect may occur through several mechanisms, including, but not limited to, direct interference with TSLP-induced JAK-STAT activation, alteration of basal expression levels of JAK-STAT proteins, potential changes in cellular responsiveness to TSLP, and regulation of feedback loops within the JAK-STAT pathway that regulate TSLP expression.

[0056] Furthermore, the effects of the compound on the JAK-STAT pathway extend beyond TSLP regulation and can potentially affect other inflammatory mediators and cellular processes. This broad influence on central signaling pathways highlights the potential of the compound as a multifaceted anti-inflammatory agent capable of addressing various aspects of inflammatory conditions such as atopic dermatitis and asthma.

[0057] A comprehensive analysis of the JAK-STAT pathway components presented in these figures not only supports the proposed mechanism of action of this small molecule compound but also provides a foundation for understanding its broader effects on inflammatory signaling networks. This detailed characterization of pathway regulation can prove valuable for predicting therapeutic efficacy, identifying potential biomarkers for therapeutic response, and guiding the future optimization of compounds targeting the Rac2-TSLP-JAK-STAT axis or the development of related therapies.

[0058] The specificity of the mechanism of this small molecule compound is further exemplified in Figures 24a and 24b. These volcano plots show differential gene expression analyses comparing Th2 stimulation at various doses with a vehicle control and with Th2 stimulation and treatment with this small molecule compound. The plots demonstrate a dose-dependent response to the compound in which specific genes associated with inflammatory pathways are significantly affected.

[0059] In some cases, the reduction in TSLP expression caused by this small molecule compound may range from about 30% to about 90%, alternatively from about 40% to about 80%, and alternatively from about 50% to about 70% compared to the control or placebo. The degree of TSLP reduction may vary depending on factors such as the specific inflammatory condition being treated, the dosage of the administered compound, and individual patient characteristics.

[0060] The regulation of Rac2 activity by this small molecule compound can affect various cellular processes in addition to TSLP production. In some cases, the compound may affect immune cell migration, cytokine production, and cell adhesion. These additional effects may contribute to the compound's overall anti-inflammatory action in pathological conditions such as atopic dermatitis and asthma.

[0061] The mechanism of action described herein represents a significant advance in the treatment of inflammatory conditions by specifically targeting the Rac2-TSLP signaling axis. This approach offers several advantages over traditional broad-spectrum immunosuppressive therapies. By selectively modulating Rac2 activity, this small molecule compound achieves more precise intervention in the inflammatory cascade, resulting in potentially improved efficacy and a reduced risk of systemic side effects.

[0062] The ability of compounds to interfere with the interaction between Rac2 and RhoGDI is a key characteristic of their mechanism. This interference alters the intracellular distribution and activation status of Rac2, ultimately leading to a decrease in TSLP expression. TSLP, a cytokine crucial for the initiation and maintenance of allergic inflammation, plays a central role in pathological conditions such as atopic dermatitis and asthma. By targeting the upstream regulator Rac2, the compounds indirectly regulate TSLP levels, potentially affecting multiple downstream inflammatory pathways simultaneously.

[0063] The increased binding affinity of this small molecule compound to Rac2 compared to the related proteins Rac1 and Rac3 is important for its targeted therapeutic approach. This selectivity, which can be as high as 5.7-fold for Rac2 and 3.9-fold for Rac3 compared to Rac1, specifically regulates Rac2-dependent inflammatory processes while minimizing interference with essential cellular functions mediated by Rac1 and Rac3. This selectivity profile contributes to the compound's potential for a favorable therapeutic range and a differentiated safety profile compared to less selective anti-inflammatory agents.

[0064] The effects of the compound extend beyond TSLP regulation to affect various cell types and inflammatory mediators. Studies have shown that it can reduce the levels of serum IgE as well as other key cytokines such as IL-13 and IL-31. This broad impact on the inflammatory network suggests that the compound provides a comprehensive approach to managing complex inflammatory conditions, capable of simultaneously addressing multiple aspects of disease pathophysiology.

[0065] Furthermore, as evidenced by changes in the gene expression of pathway components such as Jak1, Jak2, Jak3, and various STAT proteins, the ability of compounds to modulate the JAK-STAT pathway provides an additional mechanism for their anti-inflammatory effects. This modulation of the central signaling pathway of inflammation further underpins the potential of the compounds as multifaceted therapeutic agents.

[0066] In summary, the novel mechanism of action of this small molecule compound, which combines selective Rac2 modulation with downstream effects on TSLP and other inflammatory mediators, represents a promising approach for the treatment of various inflammatory conditions. Its targeted nature and potential for broad anti-inflammatory effects position it as a valuable addition to the treatment of conditions such as atopic dermatitis and asthma, potentially offering improved outcomes for patients who do not respond adequately to existing therapies.

[0067] In some cases, this small molecule compound may be particularly effective in treating atopic dermatitis. Administration of a therapeutically effective dose of the compound may result in a reduction in the dermatitis severity score. The reduction in the severity score may range from about 20% to about 80%, alternatively from about 30% to about 70%, and alternatively about 50% compared to baseline or placebo treatment.

[0068] The therapeutically effective dose of this small molecule compound may vary depending on factors such as the severity of the condition, patient characteristics, and the route of administration. For oral administration, the dosage may be in the range of about 0.1 mg / kg to about 50 mg / kg body weight per day, alternatively about 1 mg / kg to about 30 mg / kg body weight per day, and alternatively about 10 mg / kg body weight per day. For topical administration, the concentration of the compound in the formulation may be in the range of about 0.01% to about 10% w / w, alternatively about 0.1% to about 5% w / w, and alternatively about 1% w / w.

[0069] This small molecule compound may be administered via various routes, including but not limited to oral, topical, intravenous, subcutaneous, or inhalation. The choice of administration route may depend on the specific inflammatory condition being treated and the desired pharmacokinetic profile. In the case of atopic dermatitis, topical administration may be preferred to maximize local effects and minimize systemic exposure.

[0070] In some cases, this small molecule compound may be used in combination with other therapeutic agents to enhance efficacy or address various aspects of inflammatory conditions. Potential combination therapies may include corticosteroids, calcineurin inhibitors, or other immunomodulators. In combination therapy, the dosage of this small molecule compound may be in the range of about 50% to about 150% of the monotherapy dose, alternatively about 75% to about 125% of the monotherapy dose, and alternatively about 100% of the monotherapy dose.

[0071] This small molecule compound may also be effective in treating other inflammatory conditions in addition to atopic dermatitis. These conditions may include, but are not limited to, asthma, allergic rhinitis, and eosinophilic esophagitis. The dosage and route of administration for these conditions can be adjusted based on the specific pathophysiology and the target tissues involved.

[0072] For the treatment of asthma, the small molecule compound may be administered by inhalation. The inhaled dose may be in the range of about 50 μg to about 1000 μg daily, alternatively about 100 μg to about 500 μg daily, and alternatively about 250 μg daily. The compound may be formulated as a dry powder inhaler, a metered-dose inhaler, or a nebulizer solution.

[0073] When treating allergic rhinitis, this small molecule compound may be administered intranasally. The intranasal dose may be in the range of about 10 μg to about 500 μg per nostril daily, alternatively about 50 μg to about 250 μg per nostril daily, and alternatively about 100 μg per nostril daily.

[0074] In the case of eosinophilic esophagitis, oral administration of this small molecule compound may be preferred. Oral doses for this condition may range from about 0.5 mg / kg to about 20 mg / kg body weight daily, alternatively from about 1 mg / kg to about 10 mg / kg body weight daily, and alternatively from about 5 mg / kg body weight.

[0075] The duration of treatment with this small molecule compound may vary depending on the nature and severity of the inflammatory condition. In the case of chronic conditions such as atopic dermatitis, long-term administration may be necessary. The duration of treatment may range from about 1 week to about 52 weeks, alternatively from about 4 weeks to about 24 weeks, and alternatively about 12 weeks. In some cases, maintenance therapy with reduced administration frequency may be necessary to maintain therapeutic effects.

[0076] The efficacy of this small molecule compound in treating inflammatory conditions can be monitored through various clinical and laboratory parameters. In the case of atopic dermatitis, these parameters may include changes in dermatitis severity scores, reduction in itching, improvement in sleep quality, and reductions in inflammatory biomarkers such as serum IgE levels. A decrease in TSLP expression can serve as a pharmacodynamic indicator of therapeutic efficacy, and a reduction of at least 50% compared to baseline or placebo is considered significant.

[0077] In summary, this small molecule compound, which binds to Rac2 with increased affinity compared to Rac1 and Rac3, can provide a novel therapeutic approach for various inflammatory conditions. By modulating Rac2 activity and reducing TSLP expression, the compound can provide effective treatment for atopic dermatitis and other inflammatory disorders with the potential for improved targeting and reduced side effects compared to broader immunosuppressive therapies.

[0078] small molecule compounds

[0079] A compound of formula (I) or formula (II) is provided herein as the following formula or a pharmaceutically acceptable salt thereof:

[0080]

[0081] In the above formula,

[0082] R 1 alkyl, -CN, -NH2, -OH, -CH2OH, -CHO, -CH2R 8 , -COOH, -CONH2, -COR 8 , -N3, -SH -S(alkyl), halogen atom, aryl, heteroaryl, or R 3 Connected to form a 3- to 10-membered cyclic, heterocyclic, or heteroaromatic group, wherein the alkyl, aryl, heteroaryl, and cyclic or heterocyclic group is unsubstituted or one or more oxos or R8 Replaced with;

[0083] R 2 is oxygen or NR 9 And;

[0084] R 3 ε₀ = hydroxyl, =O, =N-OH, halogen, -SH, -S (alkyl), -NR₀ 9 R 10 , -COOH, -COOR 9 , -OCOR 9 , -CONH2, -CN, -N3, haloalkyl, or alkoxy;

[0085] R 4 , R 5 , and R 6 Each is independently hydrogen or an unsubstituted or substituted alkyl;

[0086] R 7 Silver is oxygen, nitrogen, or NR 9 And;

[0087] Each R 8 are independently hydroxyl, halogen, -SH, -S (alkyl), -NR 9 R 10 , -COOH, -COOR 9 , -OCOR 9 , -CONH2, -COR 9 , -COHR 9 R 10 , -CN, -N3, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, aryl, aralkyl, or heterocyclil;

[0088] R 9 and R 10 Each is independently hydrogen, cycloalkyl, or a C1-C5 unsubstituted or substituted alkyl;

[0089] However, the compound of chemical formula (I) is not isoandrogramolide.

[0090] In some implementations,

[0091] R 1C1-C6 alkyl, -CN, -NH2, -OH, -CH2OH, -CHO, -CH2R 8 , -COOH, -CONH2, -COR 8 , -N3, -SH -SMe, halogen atom, aryl, heteroaryl or R 3 Connected to form a 3- to 10-membered cyclic, heterocyclic, or heteroaromatic group, wherein the C1-C6 alkyl, aryl, heteroaryl, and cyclic or heterocyclic group is unsubstituted or one or more oxos or R 8 Replaced with;

[0092] R 2 is oxygen or NR 9 And;

[0093] R 3 ε₀ = hydroxyl, =O, =N-OH, halogen, -SH, -SMe, -NR₀ 9 R 10 , -COOH, -COOR 9 , -OCOR 9 , -CONH2, -CN, -N3, C1-C5 haloalkyl, or C1-C5 alkoxy;

[0094] R 4 , R 5 , and R 6 Each is independently hydrogen or a C1-C5 unsubstituted or substituted alkyl;

[0095] R 7 Silver is oxygen, nitrogen, or NR 9 And;

[0096] Each R 8 are independently hydroxyl, halogen, -SH, -SMe, -NR 9 R 10 , -COOH, -COOR 9 , -OCOR 9 , -CONH2, -COR 9 , -COHR 9 R 10, -CN, -N3, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, C1-C5 alkoxy, aryl, aralkyl, or heterocyclil;

[0097] R 9 and R 10 Each is independently hydrogen, a C3-C8 cycloalkyl, or a C1-C5 alkyl, wherein each C1-C5 alkyl is optionally substituted with a phenyl, and each phenyl is optionally substituted with one or more groups selected from C1-C5 alkyl and a halogen;

[0098] However, the compound of chemical formula (I) is not isoandrogramolide.

[0099] In some implementations,

[0100] R 1 alkyl, -CN, -NH2, -OH, -CH2OH, -CHO, -CH2R 8 , -COOH, -CONH2, -COR 8 , -N3, -COR 8 , -SH -S(alkyl), halogen, aryl, heteroaryl, or R 3 Connected to form a 3- to 10-membered cyclic or heterocyclic group, wherein the alkyl, aryl, heteroaryl, and cyclic or heterocyclic group are unsubstituted or one or more of R 8 Replaced with;

[0101] R 2 is oxygen or NR 9 And;

[0102] R 3 is a hydroxyl, =O, halogen, -SH, -S (alkyl), -NR 9 R 10 , -COOH, -COOR 9 , -CONH2, -CN, -N3, haloalkyl, or alkoxy;

[0103] R 4 , R 5 , and R 6Each is independently hydrogen or an unsubstituted or substituted alkyl;

[0104] R 7 Silver is oxygen, nitrogen, or NR 9 And;

[0105] Each R 8 are independently hydroxyl, halogen, -SH, -S (alkyl), -NR 9 R 10 , -COOH, -COOR 9 , -CONH2, -COR 9 , -COHR 9 R 10 , -CN, -N3, alkyl, haloalkyl, alkenyl, alkynyl, alkoxy, aryl, or aralkyl;

[0106] R 9 and R 10 Each is independently hydrogen or an unsubstituted or substituted alkyl;

[0107] However, the compound of chemical formula (I) is not isoandrogramolide.

[0108] In some implementations,

[0109] R 1 C1-C6 alkyl, -CN, -NH2, -OH, -CH2OH, -CHO, -CH2R 8 , -COOH, -CONH2, -COR 8 , -N3, -COR 8 , -SH -SMe, halogen atom, aryl, heteroaryl or R 3 Connected to form a 3- to 10-membered cyclic or heterocyclic group, and the C1-C6 alkyl, aryl, heteroaryl, and cyclic or heterocyclic groups are unsubstituted or one or more of R 8 Replaced with;

[0110] R 2 is oxygen or NR 9 And;

[0111] R 3 is hydroxyl, =O, halogen, -SH, -SMe, -NR9 R 10 , -COOH, -COOR 9 , -CONH2, -CN, -N3, C1-C5 haloalkyl, or C1-C5 alkoxy;

[0112] R 4 , R 5 , and R 6 Each is independently hydrogen or a C1-C5 unsubstituted or substituted alkyl;

[0113] R 7 Silver is oxygen, nitrogen, or NR 9 And;

[0114] Each R 8 are independently hydroxyl, halogen, -SH, -SMe, -NR 9 R 10 , -COOH, -COOR 9 , -CONH2, -COR 9 , -COHR 9 R 10 , -CN, -N3, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, C1-C5 alkoxy, aryl, or aralkyl;

[0115] R 9 and R 10 Each is independently hydrogen or a C1-C5 unsubstituted or substituted alkyl;

[0116] However, the compound of chemical formula (I) is not isoandrogramolide.

[0117] For the purposes of this disclosure, chemical elements are identified according to the literature [Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed]. Additionally, general principles of organic chemistry are described in the literature ["Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry," 5th Ed., Ed.: Smith, MB and March, J., John Wiley & Sons, New York: 2001], the entire contents of which are incorporated herein by reference.

[0118] As described herein, the compounds of the present disclosure may optionally be substituted with one or more substituents as generally exemplified above, or as exemplified by specific classes, subclasses, and species of the present disclosure. As will be recognized by those skilled in the art, the combinations of substituents conceived by the present disclosure are combinations that result in the formation of stable or chemically feasible compounds. As used herein, the term “stable” refers to a compound that does not substantially change when subjected to conditions that enable its production, detection, and preferably its recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, a stable compound or a chemically feasible compound is a compound that does not substantially change when maintained at a temperature of 40°C or lower for at least one week, optionally for at least four weeks, optionally for at least eight weeks, or optionally for at least twelve weeks in the absence of moisture or other chemically reactive conditions.

[0119] The phrase “optionally substituted” may be used interchangeably with the phrase “substituted or unsubstituted.” Generally, whether or not the term “optionally” precedes it, the term “substituted” refers to the replacement of a hydrogen radical of a given structure with a radical of a specific substituent. Specific substituents are described in the definition above and in the description of the compounds and examples thereof below. Unless otherwise indicated, the optionally substituted group may have a substituent at each substitutable position of the group, and if more than one position in any given structure may be substituted with more than one substituent selected from the specific group, the substituents may be the same or different at all positions. A ring substituent, such as a heterocycloalkyl, may be bonded to another ring, such as a cycloalkyl, to form a spiro-bicyclic ring system, for example, in which both rings share a single common atom. As will be recognized by those skilled in the art, the combinations of substituents conceived by this disclosure are combinations that result in the formation of a stable or chemically feasible compound.

[0120] As used herein, the phrase “maximum” refers to any integer equal to or less than 0 or the number following the phrase. For example, “maximum 4” means any one of 0, 1, 2, 3, and 4.

[0121] As used herein, the terms "aliphatic," "aliphatic group," or "alkyl" refer to a straight chain (which is completely saturated or contains one or more unsaturated units) in other wordsIt means a hydrocarbon chain that is unbranched or branched, substituted, or unsubstituted. Unless otherwise specified, the aliphatic group contains 1 to 20 aliphatic carbon atoms. In some embodiments, the aliphatic group contains 1 to 10 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1 to 8 aliphatic carbon atoms. In further other embodiments, the aliphatic group contains 1 to 6 aliphatic carbon atoms, and in further other embodiments, the aliphatic group contains 1 to 4 aliphatic carbon atoms. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted, or unsubstituted alkyl, alkenyl, or alkynyl groups.

[0122] The terms “alicyclic” or “cycloalkyl” mean a monocyclic hydrocarbon ring, or a polycyclic hydrocarbon ring system that is fully saturated or contains one or more unsaturated units but is not aromatic and has a single attachment point to the rest of the molecule. As used herein, the term “polycyclic ring system” comprises bicyclic and tricyclic 4- to 12-membered structures forming at least two rings, wherein the two rings have at least one common atom (e.g., two common atoms) comprising a fused, cross-linked, or spirocyclic ring system.

[0123] As used herein, the terms “halogen” or “halo” mean F, Cl, Br, or I. Unless otherwise specified, as used herein, the terms “heterocycle,” “heterocyclile,” “heterocycloalkyl,” or “heterocyclic” mean a non-aromatic, monocyclic, bicyclic, or tricyclic ring system in which one or more ring atoms in one or more ring members are independently selected heteroatoms. A heterocyclic ring may be saturated or may contain one or more unsaturated bonds. In some embodiments, the “heterocycle,” “heterocyclile,” “heterocycloalkyl,” or “heterocyclic” group has 3 to 14 ring members, wherein one or more ring members are heteroatoms independently selected from oxygen, sulfur, nitrogen, or phosphorus, and each ring of the ring system contains 3 to 7 ring members.

[0124] The term "heteroatom" refers to oxygen, sulfur, nitrogen, phosphorus, or silicon (any oxidized form of nitrogen, sulfur, phosphorus, or silicon; any quaternized form of basic nitrogen; a substitutable nitrogen of a heterocyclic ring, e.g., N (as in 3,4-dihydro-2H-pyrrolil), NH (as in pyrrolidinyl), or NR + It means containing (as in N-substituted pyrrolidinyl).

[0125] As used herein, the term "unsaturated" means that a moiety has one or more unsaturated units but is not aromatic.

[0126] The terms “alkoxy” or “thioalkyl” as used herein refer to an alkyl group as previously defined, attached to a main carbon chain through an oxygen (“alkoxy”) or sulfur (“thioalkyl”) atom.

[0127] The term "aryl," used alone or as part of a larger moiety as in "aralkyl," "aralkoxy," or "aryloxyalkyl," refers to a monocyclic, bicyclic, and tricyclic ring system having a total of 5 to 14 cyclic carbon atoms, wherein at least one ring in the system is aromatic and each ring in the system contains 3 to 7 cyclic carbon atoms. The term "aryl" may be used interchangeably with the term "aryl ring."

[0128] The term “heteroaryl,” used alone or as part of a larger moiety as in “heteroaralkyl” or “heteroarylalkoxy,” refers to a monocyclic, bicyclic, and tricyclic ring system having a total of 5 to 14 ring members, wherein at least one ring in the system is aromatic, at least one ring in the system contains one or more heteroatoms, and each ring in the system contains 3 to 7 ring members. The term “heteroaryl” may be used interchangeably with the term “heteroaryl ring” or the term “heteroaromatic.”

[0129] "D" and "d" both refer to deuterium.

[0130] In the structure, dotted lines represent connections that may or may not be present. For example, represents a bond that is a single bond or a double bond.

[0131] Unless otherwise noted, the structures described herein also mean all isomer forms of the structure (e.g., enantiomers, diastereomers, and geometric (or stereomorphic) forms); e.g., R and S coordinations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) stereomorphic isomers. Thus, not only the single stereochemical isomers of the compound, but also enantiomers, diastereomers, and geometric (or stereomorphic) mixtures are within the scope of this disclosure. Unless otherwise noted, all tautomeric forms of the compound of formula (I) of this disclosure are within the scope of this disclosure. Thus, tautomeric forms of the compound of formula (I) are included within the scope of this disclosure. The structures also include, where appropriate, zwitterionic forms of the compound or salt of formula (I).

[0132] Additionally, unless otherwise stated, the structures described herein are also meant to include compounds that differ only in the presence of one or more isotope-rich or isotope-labeled atoms. The isotope-labeled compounds may have one or more atoms replaced by atoms having atomic masses or mass numbers that are commonly found in nature. Examples of isotopes present in the compounds of formula (I) include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, e.g., 2H, 3H, 13C, 14C, 15N, 18O, 17O, 35S, and 18F, but not limited to. Specific isotope-labeled compounds of formula (I) are useful not only as therapeutic agents but also as analytical tools or probes in drug and / or substrate tissue distribution assays. In one aspect of the present disclosure, tritium (e.g., 3H) and carbon-14 (e.g., 14C) isotopes are useful considering their ease of detection. In another aspect of the present disclosure, replacing one or more hydrogen atoms with heavier isotopes, such as deuterium (e.g., 2H), may provide specific therapeutic benefits.

[0133] In a specific embodiment, the compound of formula (I) is formula (Ia) or a pharmaceutically acceptable salt thereof, and the compound of formula (II) is formula (IIa) or a pharmaceutically acceptable salt thereof:

[0134]

[0135] In the above formula,

[0136] R 1 C1-C6 alkyl, -CN, -NH2, -OH, -CH2OH, CHO, -CH2R 8 , -COOH, -CONH2, -COR 8 , -N3, -COR 8, -SH -SMe, a halogen atom, aryl, or heteroaryl, wherein the C1-C6 alkyl, aryl, and heteroaryl are unsubstituted or one or more R 8 Replaced with;

[0137] R 2 is oxygen or NR 9 And;

[0138] R 7 Silver is oxygen, nitrogen, or NR 9 And;

[0139] Each R 8 are independently hydroxyl, halogen, -SH, -SMe, -NR 9 R 10 , -COOH, -COOR 9 , -CONH2, -COR 9 , -COHR 9 R 10 , -CN, -N3, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, C1-C5 alkoxy, aryl, or aralkyl;

[0140] R 9 and R 10 Each is independently hydrogen or a C1-C5 unsubstituted or substituted alkyl;

[0141] However, the compound of chemical formula (Ia) is not isoandrogramolide.

[0142] In some embodiments, the compound of formula (Ia) is formula (Iaa) or a pharmaceutically acceptable salt thereof, and the compound of formula (IIa) is formula (IIaa) or a pharmaceutically acceptable salt thereof:

[0143]

[0144]

[0145] In the above formula,

[0146] R 1-1 -CN, -NH2, -OH, -COOH, CHO, -CH2R 8, -CONH2, -COR 8 , -N3, -COR 8 , -SH, -SMe, halogen atom, aryl, or heteroaryl, where the aryl and heteroaryl are unsubstituted or one or more R 8 Replaced with;

[0147] R 1-2 is C1-C6 alkyl, -CN, -NH2, -OH, -CH2OH, -COOH, CHO, -CH2R 8 , -CONH2, -COR 8 , -N3, -COR 8 , -SH, -SMe, halogen atom, aryl, or heteroaryl, wherein C1-C6 alkyl, aryl, and heteroaryl are unsubstituted or one or more R 8 Replaced with;

[0148] Each R 8 are independently hydroxyl, halogen, -SH, -SMe, -NR 9 R 10 , -COOH, -COOR 9 , -CONH2, -COR 9 , -COHR 9 R 10 , -CN, -N3, C1-C5 alkyl, C1-C5 haloalkyl, C2-C5 alkenyl, C2-C5 alkynyl, C1-C5 alkoxy, aryl, or aralkyl;

[0149] R 9 and R 10 Each is independently hydrogen or a C1-C5 unsubstituted or substituted alkyl.

[0150] In certain embodiments, the compound has the chemical formula (Iaa). In these and other embodiments, R 1-1 It is -CN. In an exemplary embodiment, the compound of chemical formula (Iaa) is as follows.

[0151]

[0152] In certain embodiments, the compound is of formula (IIaa). In some embodiments, the compound of formula (IIaa) is selected from the group consisting of the following formula or pharmaceutically acceptable salts or stereoisomers thereof:

[0153]

[0154]

[0155]

[0156] In the above formula,

[0157] R 9-1 It is hydrogen or methyl;

[0158] R 9-2 is methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, secondary butyl, or cyclobutyl.

[0159] In these and other embodiments, R 1-2 is -CH2OH or -CN. In some exemplary embodiments, the compound is the following formula or a pharmaceutically acceptable salt or stereoisomer thereof:

[0160] . In other exemplary embodiments, the compound is of the following formula or a pharmaceutically acceptable salt or stereoisomer thereof:

[0161] .

[0162] In various embodiments, the compound of formula (I) or formula (II) is provided as an inhibitor or modulator of one or more of the following therapeutic targets: neutrophil migration, cathepsin D (Cath D), surfactant protein C (SPC), glutathione (GSH), collagen, type I, alpha 1 (COL1A1), pyruvate carboxylase (PC), 90kD heat shock protein (Hsp90β), and transforming growth factor-beta (TGF-β1). In relation to or independently of such inhibitory or modulatory activity, the compound is useful for the treatment of diseases, disorders, and conditions including, but not limited to, fibrosis.

[0163] Pharmaceutically acceptable salts and compositions

[0164] In another aspect of the present disclosure, pharmaceutically acceptable compositions are provided, wherein the compositions comprise a compound of formula (I) or formula (II) as described herein and optionally comprise a pharmaceutically acceptable carrier, adjuvant, or vehicle. In certain embodiments, the compositions optionally further comprise one or more additional therapeutic agents.

[0165] It will also be understood that the specified compounds of the present disclosure may exist in a free form for therapeutic purposes, or, where appropriate, as pharmaceutically acceptable derivatives thereof. According to the present disclosure, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, salts of such esters, or any other adducts or derivatives that, when administered to a patient in need, may directly or indirectly provide the compounds otherwise described herein, or metabolites or residues thereof.

[0166] As used herein, the term “pharmaceuticalally acceptable salt” refers to such salt that, within the scope of appropriate medical judgment, is suitable for use in contact with human and lower animal tissues without excessive toxicity, irritation, allergic reactions, etc., and corresponds to a reasonable benefit / risk ratio. “Pharmaceuticalally acceptable salt” means any non-toxic salt or salt of an ester of the compound of the present disclosure that, when administered to a recipient, may provide, directly or indirectly, the compound of the present disclosure or its inhibitory active metabolite or residue. As used herein, the term “its inhibitory active metabolite or residue” means that its metabolite or residue is also an inhibitor or modifier of one or more of the therapeutic targets.

[0167] Pharmaceutically acceptable salts are widely known in the art. For example, pharmaceutically acceptable salts are described in the literature by SM Berge et al. [J. Pharmaceutical Sciences, 1977, 66, 1-19], the entirety of which is incorporated herein by reference. Pharmaceutically acceptable salts of Formula (I) or Formula (II) of the present disclosure include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, non-toxic acid addition salts are salts formed using inorganic acids, such as hydrochloric acid, hydrobromide, phosphoric acid, sulfuric acid, and perchloric acid, or organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by other methods used in the art, such as ion exchange. Other pharmaceutically accepted salts are adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphosulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, maleate, malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, It includes pectinates, persulfates, 3-phenylpropionate, phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerate salts, etc. Salts derived from suitable bases include alkali metals, alkaline earth metals, ammonium, and N + (C 1-4It includes alkyl)tetrasalts. The present disclosure also considers the quaternization of any basic nitrogen-containing group of the compound of formula (I) or formula (II) disclosed herein. By such quaternization, water or oil-soluble or dispersible products may be obtained. Representative alkali metal salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Additionally, pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfons, and aryl sulfons, where appropriate.

[0168] As described herein, the pharmaceutically acceptable compositions of the present disclosure additionally comprise pharmaceutically acceptable carriers, adjuvants, or vehicles, which, when used herein, comprise any and all solvents, diluents, or other liquid vehicles, dispersion or suspension aids, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc. suitable for a specific dosage form desired. The literature [Remington's Pharmaceutical Sciences, Sixteenth Edition, EW Martin (Mack Publishing Co., Easton, Pa., 1980)] discloses various carriers used in the formulation of pharmaceutically acceptable compositions and known techniques for their manufacture. Unless any conventional carrier is incompatible with the compound of Formula (I) or Formula (II) of the present disclosure by, for example, producing any undesirable biological effect or otherwise interacting in a harmful manner with any other component(s) of a pharmaceutically acceptable composition, its use is deemed to be within the scope of the present disclosure.Examples of some substances that can serve as pharmaceutically acceptable carriers include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, or potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes, protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-block copolymers, wool fats, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; Excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols such as propylene glycol or polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic brine; Ringer's solution; ethyl alcohol, and phosphate buffer solution, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, are included but not limited thereto, as well as coloring agents, release agents, coating agents, sweeteners, flavoring and fragrance agents, preservatives, and antioxidants may also be present in the composition at the discretion of the manufacturer.

[0169] In another aspect, the present disclosure features a pharmaceutical composition comprising a compound of the present disclosure and a pharmaceutically acceptable carrier.

[0170] In another aspect, the present disclosure features a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers or vehicles.

[0171] The compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof used in the composition to be administered may be obtained from any commercially available source or from any method or technique known in the art. For example, in some variations, the compound of formula (I) or formula (II) may be isolated from a natural source. In other variations, the compound of formula (I) or formula (II) may be synthesized according to any method known in the art.

[0172] In various embodiments, the composition comprises a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof, and optionally consists essentially of a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof. As used herein, the phrase “consists essentially” generally includes the elements / components specifically mentioned for a particular embodiment. Additionally, the phrase “consists essentially” generally encompasses and allows for the presence of additional or optional elements / components that do not substantially affect the basic and / or novel properties of the particular embodiment. In a particular embodiment, “consists essentially” allows for the presence of ≤10, ≤5, or ≤1 weight percent (weight%) of the additional or optional components based on the total weight of the composition.

[0173] In various embodiments, the composition comprises one or more pharmaceutically acceptable additives that are inert components. Examples of inert components include, but are not limited to, excipients, e.g., diluents and binders, granulators, lubricants (or flow aids), fillers, lubricants, preservatives, stabilizers, coatings, disintegrants, fragrances, pigments, preservatives, solvents (e.g., alcohols), and combinations thereof. When used to form a composition, the inert component(s) may be used in varying amounts and may be combined with a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof to form a composition suitable for topical application to human or animal skin. It should be further understood that the amount of active agent described herein may be normalized to 100 parts by weight of the composition to account for the presence of the inert component (when used).

[0174] Optionally, the composition may include one or more additional ingredients, such as additives. Suitable additives include, but are not limited to, humectants, emollients, emulsifiers, surfactants, oils, extracts, skin protectants, disinfectants, preservatives, drugs and pharmaceutical raw materials, analgesic compounds, antineurolytic compounds, antioxidants, blood circulation promoters, antidepressant compounds, anti-anxiety compounds, anti-stress compounds, sunscreens, insect repellents, preservatives, exfoliants, fragrances, colorants, fillers, solvents, vehicles, carriers, other types of additives known to those skilled in the art, and combinations thereof. These additives may be used alone or in combination. Generally, optional additives may be any type used in pharmaceutical, functional medicines, personal care products, and cosmetic products.

[0175] Examples of such carrier components include oils, fats, waxes, surfactants, moisturizers, thickeners, antioxidants, viscosity stabilizers, chelating agents, buffers, preservatives, perfumes, dyes, lower alkanols, etc. If desired, additional components, e.g., anti-inflammatory agents, antibacterial agents, antifungal agents, disinfectants, vitamins, sunscreens, antibiotics, skin whitening agents, healing promoters / fibroblast proliferation compounds, neuromuscular blockers, sunscreens, or other anti-acne agents may be incorporated into the composition.

[0176] Examples of oils as carrier agents include fats and oils such as olive oil and hydrogenated oil; waxes such as beeswax and lanolin; hydrocarbons such as liquid paraffin, ceresin, and squalene; fatty acids such as stearic acid and oleic acid; alcohols such as cetyl alcohol, stearyl alcohol, lanolin alcohol, and hexadecanol; and esters such as isopropyl myristate, isopropyl palmitate, and butyl stearate. Examples of surfactants as carrier agents include anionic surfactants, such as sodium stearate, sodium cetyl sulfate, polyoxyethylene lauryl ether phosphate, and sodium N-acyl glutamate; cationic surfactants, such as stearyldimethylbenzylammonium chloride and stearyltrimethylammonium chloride; amphoteric surfactants, such as alkylaminoethylglycine hydrochloride solution and lecithin; and nonionic surfactants, such as glycerin monostearate, sorbitan monostearate, sucrose fatty acid ester, propylene glycol monostearate, polyoxyethylene oleyl ether, polyethylene glycol monostearate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene coconut fatty acid monoethanolarnide, polyoxypropylene glycol (e.g., a substance sold under the trade name "Pluronic"), polyoxyethylene castor oil, and polyoxyethylene lanolin may be mentioned. Examples of wetting agents as carrier agents include glycerin, 1,3-butylene glycol, and propylene glycol; examples of lower alcohols include ethanol and isopropanol; and examples of thickening agents include xanthan gum, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyethylene glycol, and sodium carboxymethyl cellulose.Examples of antioxidants include butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate, citric acid, ethoxyquin, alpha-lipoic acid, vitamin C, vitamin E, coenzyme Q-10, and idebenone; plant-based antioxidants include carotenoids, such as lycopene; flavonoids, such as silylmarin (milk thistle), silylbin, silyldianin, silylcristin; soybeans (isoflavin), grape seed extract; polyphenols, such as green tea extract, rosmarinic acid (rosemary), hypersin (St. John's wort), oleurofin (olive leaf), curcumin (turmeric root), tetrahydrocurcumin, and picogenol (marine bark pine). Examples of anti-inflammatory agents include anti-inflammatory herbal medicines such as allantoin, aloe vera, ginkgo biloba, and green tea (also considered an antioxidant). Examples of skin bleaching agents are hydroquinone or kojic acid. Examples of healing promoters / fibroblast proliferation compounds include copper peptide or palmitoyl-pentapeptide (pal-KTTKS). Examples of neuromuscular blocking agents such as acetyl hexapeptide 3 (argiclean) or dimethylaminoethanol. Examples of chelating agents include disodium edetate and ethanehydroxy diphosphate. Examples of buffers acting as carrier agents include citric acid, sodium citrate, boric acid, borax, and disodium hydrogen phosphate; examples of preservatives are methyl parahydroxybenzoate, ethyl parahydroxybenzoate, dehydroacetic acid, salicylic acid, and benzoic acid.

[0177] It should be understood that specific components or additives may be classified under different technical terms, and that the classification of components or additives under such terms does not imply that they are limited to their function. When used, additives or additives may be present in the composition in varying amounts. Additional components for optional use in the composition are described in U.S. Patent No. 5,747,006 of Dornoff et al., U.S. Patents No. 5,980,904, 6,994,874, 7,060,304, 7,247,321, and 7,364,759 of Leverett et al., the entirety of which is incorporated herein by reference, and U.S. Publication No. 2017 / 0252293 of Brumbaugh et al.

[0178] The composition may be prepared using various methods. For example, the active agent of the composition, and optionally one or more inerts, may be mixed or blended and compressed or compounded using various techniques understood in the art. The composition of the present disclosure is not limited to a specific order of manufacturing steps or methods.

[0179] The composition may be in various forms. Examples of suitable forms include solids, gels, and liquids. For example, the composition may be formulated for application as a gel, cream, lotion, pomade, mousse, powder, or foam for application to the skin of a subject. In another example, the composition may be formulated for spraying onto the skin of a subject. The composition may be formulated to be sprayed as an aerosol spray or pump spray. In yet another example, the composition may be formulated for application using a pre-moistened toe towel. In yet another example, the composition may be formulated as a solid to be rubbed onto the skin of a subject. In yet another example, the composition is formulated for delivery via a patch attached to the skin of a subject.

[0180] The composition may comprise a compound of formula (I) or a pharmaceutically acceptable salt thereof in an amount of at least about 0.375% w / w, at least about 0.75% w / w, at least about 1% w / w, at least about 1.5% w / w, at least about 3% w / w, at least 4% w / w, at least 5% w / w, at least 6% w / w, at least 7% w / w, at least 8% w / w, at least 9% w / w, at least 10% w / w, at least 11% w / w, at least 12% w / w, at least 13% w / w, at least 14% w / w, at least 15% w / w, at least 16% w / w, at least 17% w / w, at least 18% w / w, at least 19% w / w, at least 20% w / w, or even greater than that. In certain variations, the composition comprises about 0.375%, about 0.75%, about 1%, about 1.5%, about 3% w / w, about 4% w / w, about 5% w / w, about 6% w / w, about 7% w / w, about 8% w / w, about 9% w / w, about 10% w / w, about 11% w / w, about 12% w / w, about 13% w / w, about 14% w / w, about 15% w / w, about 16% w / w, about 17% w / w, about 18% w / w, about 19% w / w, about 20% w / w, or even more than that of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0181] In a specific variation, the composition is about 0.1% to about 20% w / w, about 0.1% to about 15% w / w, about 0.1% to about 5% w / w, about 0.1% to about 3% w / w, about 0.1% to about 1.5% w / w, about 0.1% to about 1% w / w, about 0.1% to about 0.75% w / w, about 0.1% to about 0.375% w / w, about 0.375% to about 5% w / w, about 0.375% to about 3% w / w, about 0.375% to about 1.5% w / w, about 0.375% to about 1% w / w, about 0.375% to about 0.75% w / w, about 0.75% to about 5% w / w, about It comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof in an amount of 0.75% to about 3% w / w, about 0.75% to about 1.5% w / w, about 0.75% to about 1% w / w, about 1% to about 5% w / w, about 1% to about 3% w / w, about 1% to about 1.5% w / w, about 1.5% to about 5% w / w, about 1.5% to about 3% w / w, about 3% to about 5% w / w, about 1% to about 15% w / w, about 2% to about 10% w / w, about 4% to about 8% w / w, about 5% to about 15% w / w, or about 8% to about 12% w / w.

[0182] In another embodiment, a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof, is present in the composition and has a purity of at least about 50% w / w, at least about 60% w / w, at least about 70% w / w, at least about 75% w / w, at least about 80% w / w, at least about 90% w / w, at least about 95% w / w, at least about 96% w / w, at least about 97% w / w, at least about 98% w / w, at least about 99% w / w, at least about 99.9% w / w, at least about 99.99% w / w, or at least about 99.999% w / w; or about 100% w / w.

[0183] In other variations of the foregoing, the composition further comprises one or more additional ingredients, such as fragrances, colorants, and / or excipients.

[0184] Uses of compounds, pharmaceutically acceptable salts, and compositions

[0185] These pharmaceutically acceptable compositions are useful for treating or reducing the severity of various diseases, disorders, or conditions, including but not limited to fibrosis. In various embodiments, the pharmaceutically acceptable compositions are one or more inhibitors or modulators of the following therapeutic targets: neutrophil migration, cathepsin D (Cath D), surfactant protein C (SPC), glutathione (GSH), collagen, type I, alpha 1 (COL1A1), pyruvate carboxylase (PC), 90kD heat shock protein (Hsp90β), and transforming growth factor-beta (TGF-β1), and are thus useful for treating or reducing the severity of the various diseases, disorders, or conditions described herein.

[0186] In various embodiments, the terms “inhibit” and “modulate” are used interchangeably to refer to neutrophil migration modulators comprising agents that reduce or inhibit biological activity, e.g., agents that inhibit the activity of a therapeutic target, e.g., modulators having any combination of the structural and / or functional properties disclosed herein.

[0187] A method for treating, improving, or preventing a neutrophil-related condition or disease is provided herein, comprising the step of administering a therapeutically effective amount of a compound of formula (I) or formula (II). In various embodiments, the condition or disease is selected from the group consisting of a cardiovascular condition or disease, an endocrine condition or disease, a nervous system condition or disease, a gastrointestinal condition or disease, a dermatological condition or disease, or a combination thereof.

[0188] A method for treating, improving, or preventing a neutrophil-related condition or disease is also provided herein, comprising the step of administering a therapeutically effective amount of a pharmaceutical composition according to the present invention. In various embodiments, the condition or disease is selected from the group consisting of a cardiovascular condition or disease, an endocrine condition or disease, a nervous system condition or disease, a gastrointestinal condition or disease, a dermatological condition or disease, or a combination thereof.

[0189] Pharmaceutical compositions comprising a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt or stereoisomer thereof, for use in a method of treating, improving, or preventing a neutrophil-related condition or disease are also provided herein. In various embodiments, the condition or disease is selected from the group consisting of a cardiovascular condition or disease, an endocrine condition or disease, a nervous system condition or disease, a gastrointestinal condition or disease, a dermatological condition or disease, or a combination thereof.

[0190] The use of a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt or stereoisomer thereof, in the manufacture of a medicine for use in treating, improving, or preventing a fibrotic condition or disease is also provided herein. In various embodiments, the condition or disease is selected from the group consisting of a cardiovascular condition or disease, an endocrine condition or disease, a nervous system condition or disease, a gastrointestinal condition or disease, a dermatological condition or disease, or a combination thereof.

[0191] A method for treating, improving, or preventing a fibrotic condition or disease is provided herein, comprising the step of administering a therapeutically effective amount of a compound of formula (I) or formula (II). In various embodiments, the condition or disease is idiopathic pulmonary fibrosis (IPF).

[0192] A method for treating, improving, or preventing a fibrotic condition or disease is also provided herein, comprising the step of administering a therapeutically effective amount of a pharmaceutical composition according to the present invention. In various embodiments, the condition or disease is idiopathic pulmonary fibrosis (IPF).

[0193] Pharmaceutical compositions comprising a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt or stereoisomer thereof, for use in a method of treating, improving, or preventing a fibrotic condition or disease are also provided herein. In various embodiments, the condition or disease is idiopathic pulmonary fibrosis (IPF).

[0194] The use of a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt or stereoisomer thereof, in the manufacture of a medicine for use in treating, improving, or preventing a fibrotic condition or disease is also provided herein. In various embodiments, the condition or disease is idiopathic pulmonary fibrosis (IPF).

[0195] A method for treating, improving, or preventing a dermatological condition or disease is provided herein, comprising the step of administering a therapeutically effective amount of a compound of formula (I) or formula (II). In various embodiments, the condition or disease is atopic dermatitis.

[0196] A method for treating, improving, or preventing a dermatological condition or disease is also provided herein, comprising the step of administering a therapeutically effective amount of a pharmaceutical composition according to the present invention. In various embodiments, the condition or disease is atopic dermatitis.

[0197] A pharmaceutical composition comprising a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt or stereoisomer thereof, for use in a method of treating, improving, or preventing a dermatological condition or disease is also provided herein. In various embodiments, the condition or disease is atopic dermatitis.

[0198] The use of a compound of formula (I) or formula (II), or a pharmaceutically acceptable salt or stereoisomer thereof, in the manufacture of a medicine for use in treating, improving, or preventing a dermatological condition or disease is also provided herein. In various embodiments, the condition or disease is atopic dermatitis.

[0199] In some embodiments, “treating” or “treatment” refers to both curative and disease-modifying treatments, as well as treatment of patients who are at risk of developing or suspected of having the disease, as well as patients diagnosed with or with the disease or medical condition, and includes the suppression of clinical recurrence. In these and other embodiments, “treating” or “treatment” refers to obtaining a beneficial or desired result, e.g., clinical outcome, in a subject, including (1) improvement of one or more symptoms caused by or associated with the disease, disorder, or condition; (2) reduction in the degree of the disease, disorder, or condition; (3) slowing or stopping the development or progression of one or more symptoms caused by or associated with the disease, disorder, or condition (e.g., stabilization of the disease, disorder, or condition); and (4) alleviation of the disease by causing improvement of one or more clinical symptoms (e.g., improvement of the disease state, enhancement of another medicinal effect, delay or cessation of disease progression, and / or increase in quality of life).

[0200] In certain embodiments, the compound of formula (I) or formula (II), or a pharmaceutically acceptable salt thereof, has an IC of less than 50 micromolar (μM), optionally less than 40 μM, optionally less than 30 μM, optionally less than 20 μM, optionally less than 10 μM, or optionally less than 1 μM. 50 One or more of the therapeutic targets were inhibited or modified. Alternatively, the compound of formula (I) or a pharmaceutically acceptable salt thereof has an IC50 of about 0.01 μM to about 100 μM, optionally about 0.1 μM to about 75 μM, optionally about 1 μM to about 50 μM, optionally about 1 μM to about 25 μM, or optionally about 1 μM to about 10 μM. 50 It inhibits or modulates one or more therapeutic targets.

[0201] In some embodiments, the method comprises the step of administering a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof in excipients to a subject to form a composition. The composition may comprise 10 to 500 mg, or 10 to 800 mg, or 20 to 1,000 mg, or 30 to 1,200 mg, or 50 to 1,500 mg, or 100 to 2,000 mg, or even more than that of a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof in a single dose unit. In a different view, at least 20 wt% of the composition, or at least 30 wt%, or at least 40 wt%, or at least 50 wt%, or at least 60 wt%, or at least 70 wt%, or at least 80 wt%, or at least 90 wt%, will be a compound of formula (I) or a pharmaceutically acceptable salt thereof. Consequently, a preferred oral single dose unit (or recommended daily dose) will be 20 to 200 mg, or 40 to 400 mg, or 60 to 600 mg, or 80 to 800 mg, or 100 to 1,000 mg, or 200 to 2,000 mg, and in some cases much higher.

[0202] As used herein, the term “administering” of a pharmaceutical composition or drug refers to both direct and indirect administration of the pharmaceutical composition or drug, wherein direct administration of the pharmaceutical composition or drug is typically performed by a healthcare professional (e.g., physician, nurse, etc.), and indirect administration includes the step of providing or making available to a healthcare professional the pharmaceutical composition or drug for direct administration (e.g., via injection, infusion, oral delivery, topical delivery, etc.).

[0203] In some variations, the method comprises the step of forming a composition by administering a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof to a subject in a petroleum-based vehicle, e.g., petroleum jelly. The composition comprises at least about 0.375% w / w, at least about 0.75% w / w, at least about 1% w / w, at least about 1.5% w / w, at least about 3% w / w, at least 4% w / w, at least 5% w / w, at least 6% w / w, at least 7% w / w, at least 8% w / w, at least 9% w / w, at least 10% w / w, at least 11% w / w, at least 12% w / w, at least 13% w / w, at least 14% w / w, at least 15% w / w, at least 16% w / w, at least 17% w / w, at least 18% w / w, at least 19% w / w, at least 20% w / w, or even greater than that of formula (I) or a pharmaceutically acceptable salt thereof. In a specific variation, the composition comprises about 0.375%, about 0.75%, about 1%, about 1.5%, about 3% w / w, about 4% w / w, about 5% w / w, about 6% w / w, about 7% w / w, about 8% w / w, about 9% w / w, about 10% w / w, about 11% w / w, about 12% w / w, about 13% w / w, about 14% w / w, about 15% w / w, about 16% w / w, about 17% w / w, about 18% w / w, about 19% w / w, about 20% w / w, or even more than that of a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof.

[0204] In various embodiments, the amount of composition administered to a subject is the amount of the compound of formula (I) or formula (II) or the pharmaceutically acceptable salt thereof (square centimeters (cm²)). 2 It is based on milligrams (mg) per skin area. The composition is at least 0.01 mg / cm² 2 (skin cm 2 mg of the sugar compound), at least 0.02 mg / cm²2 . At least 0.03 mg / cm² 2 . At least 0.04 mg / cm² 2 . At least 0.05 mg / cm² 2 . At least 0.06 mg / cm² 2 . At least 0.07 mg / cm² 2 . At least 0.08 mg / cm² 2 . At least 0.09 mg / cm² 2 . At least 0.1 mg / cm² 2 . At least 0.12 mg / cm² 2 . At least 0.14 mg / cm² 2 . At least 0.16 mg / cm² 2 . At least 0.18 mg / cm² 2 . At least 0.2 mg / cm² 2 . At least 0.22 mg / cm² 2 . At least 0.24 mg / cm² 2 . At least 0.26 mg / cm² 2 At least 0.28 mg / cm² 2 . At least 0.3 mg / cm² 2 . At least 0.32 mg / cm² 2 . At least 0.34 mg / cm² 2 . At least 0.36 mg / cm² 2 . At least 0.3 8 mg / cm² 2 . At least 0.4 mg / cm² 2 , or even an amount exceeding that may be administered to the subject. The composition is about 0.01 mg / cm² 2 to about 0.4 mg / cm² 2 , approximately 0.05 mg / cm² 2 to about 0.3 mg / cm² 2 , approximately 0.1 mg / cm² 2 to about 3 mg / cm² 2 , or about 0.15 mg / cm² 2 to about 0.25 mg / cm² 2 It can be administered to the subject in an amount of

[0205] The composition may be applied daily, multiple times a day, or in any suitable manner as needed to achieve the desired result. In the method of the present disclosure, the frequency of administration (e.g., topical application) may vary depending on several factors, including the severity of the condition or symptoms, to achieve the same or desired level of relief from the symptoms. Generally, the regimen involves the application of the composition once or twice daily to include a morning administration and / or an evening administration. The amount and / or frequency of application of the composition may vary depending on several factors, including the level of the desired result and the specific composition. In some embodiments, the composition is administered once daily. In other embodiments, the composition is administered twice daily. In further other embodiments, the composition is administered three times daily. In further other embodiments, the composition is administered four times daily. However, it should be understood that the composition may be administered more than four times daily.

[0206] In some variations, the composition is administered for at least about 1 week, at least about 2 weeks, at least about 3 weeks, at least about 4 weeks, at least about 8 weeks, at least about 12 weeks, at least about 16 weeks, at least about 24 weeks, at least about 32 weeks, at least about 40 weeks, at least about 48 weeks, or at least about 1 year. In certain variations, the composition is administered for about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 8 weeks, about 12 weeks, about 16 weeks, about 24 weeks, about 32 weeks, about 40 weeks, about 48 weeks, or about 1 year. In one variant, the composition is administered for about 1 week to about 1 year, about 4 weeks to about 8 weeks, about 4 weeks to about 12 weeks, about 4 weeks to about 16 weeks, about 4 weeks to about 24 weeks, about 4 weeks to about 32 weeks, about 4 weeks to about 40 weeks, about 4 weeks to about 48 weeks, or about 4 weeks to about 1 year.

[0207] A compound of formula (I) or formula (II), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof may be administered as described herein. In some embodiments, the composition is administered topically to a subject. In other embodiments, the composition is administered orally.

[0208] In another aspect, the present disclosure features a method for inhibiting a therapeutic target, e.g., neutrophil migration, in a subject, comprising the step of administering to the subject a compound of formula (I) or formula (II), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof. In connection with or independently thereof, the present compound is useful for the treatment of diseases, disorders, and conditions, including but not limited to fibrotic diseases.

[0209] As used herein, the term "fibrosis" refers to a disease, disorder, or condition characterized wholly or partially by the excessive production of fibrous material, including the excessive production of fibrous material within the extracellular matrix, or by the replacement of normal tissue components by the abnormal and excessive accumulation of matrix-related components, i.e., non-functional or abnormal components. Acute and chronic, clinical or subclinical presentations of fibrotic diseases, disorders, or conditions are all possible, and all may be accompanied by clearly visible fibrotic biology or pathology.

[0210] Examples of fibrotic diseases, disorders, and conditions include systemic sclerosis, multifocal fibrosclerosis, nephrogenic systemic fibrosis, scleroderma (including Morpea, generalized Morpea, or linear scleroderma), sclerosing graft-versus-host disease, renal fibrosis (including glomerulosclerosis, renal tubulointerstitial fibrosis, progressive renal disease, or diabetic nephropathy), cardiac fibrosis (e.g., cardiomyopathy), pulmonary fibrosis (e.g., pulmonary fibrosis, glomerulosclerotic pulmonary fibrosis, idiopathic pulmonary fibrosis, silicosis, asbestosis, interstitial lung disease, interstitial fibrotic lung disease, and chemotherapy / radiation-induced pulmonary fibrosis), oral fibrosis, endocardial fibrosis, deltoid fibrosis, pancreatitis, inflammatory bowel disease, Crohn's disease, nodular fasciitis, eosinophilic fasciitis, general fibrosis syndrome characterized by the replacement of normal muscle tissue with varying degrees of fibrous tissue, retroperitoneal fibrosis, and hepatic fibrosis. Liver cirrhosis, chronic renal failure; myelofibrosis (myelofibrosis), drug-induced ergotitis, myelodysplastic syndrome, myeloproliferative syndrome, gynecological cancer, Kaposi's sarcoma, leprosy, collagenous colitis, acute fibrosis, and organ-specific fibrosis, etc.

[0211] For example, pulmonary fibrosis, pulmonary hypertension, cystic fibrosis, asthma, chronic obstructive pulmonary disease, hepatic fibrosis, renal fibrosis, NASH, and similar conditions are examples of organ-specific fibrotic diseases. The extracellular matrix is ​​abnormally and / or excessively deposited in many fibrotic diseases, disorders, and conditions. Fibrosis can be caused by surgery, the post-wound healing process, or underlying conditions. It may also be associated with inflammation. Uncontrolled fibrosis can lead to scarring, which is a deterioration of the structure of underlying organs or tissues.

[0212] Compounds of formula (I) or formula (II) may be used in some embodiments to treat or prevent pulmonary fibrosis. The list of pulmonary fibrosis conditions includes pulmonary fibrosis, pulmonary hypertension, chronic obstructive pulmonary disease (COPD), asthma, silicosis, asbestosis, cystic fibrosis, familial pulmonary fibrosis, sarcoidosis, carbon fibrosis, hypersensitivity pneumoconiosis, pulmonary fibrosis induced by inhalation of inorganic dust, and induced pulmonary fibrosis.

[0213] The progressive scarring of lung tissue, the excessive accumulation of extracellular matrix proteins, and abnormal alveolar structure are characteristics of pulmonary fibrosis. Stiff and swollen tissue makes it difficult for the lungs to function properly, causing respiratory problems such as shortness of breath and can be fatal. Acute lung injury, viral infections, toxic exposure, radiation exposure, chronic diseases, drugs, or idiopathic (unknown underlying cause) conditions can all induce pulmonary fibrosis.

[0214] The characteristic signs of idiopathic pulmonary fibrosis are diffuse peripheral lung scarring and small vesicles (called vesicles) located near the outer surface of the lungs, often around the base of the lungs. The course of idiopathic pulmonary fibrosis is often slow and relentless. Initially, patients often suffer from a dry and mysterious cough. Next, dyspnea (shortness of breath) develops and worsens over time as a result of reduced activity. Shortness of breath eventually limits all activity and can occur even while sitting quietly. In fewer cases, the fibrosis progresses rapidly, potentially causing dyspnea and disability within weeks to months of the onset of the disease. The term Hamman-Rich syndrome has been used to describe this type of pulmonary fibrosis.

[0215] An increase in blood pressure in the pulmonary vascular system, including the pulmonary arteries, pulmonary veins, and / or pulmonary capillaries, is a sign of pulmonary hypertension. The right ventricle of the heart becomes strained and enlarged due to abnormally high pressure. Heart failure can occur as the right ventricle loses the power and ability to pump adequate blood to the lungs over time. Chronic liver disease and cirrhosis, rheumatic diseases such as scleroderma or systemic lupus erythematosus (lupus), and lung diseases such as tumors, emphysema, chronic obstructive pulmonary disease (COPD), and pulmonary fibrosis can all contribute to pulmonary hypertension. Pulmonary fibrosis can cause pulmonary hypertension by narrowing the pulmonary vascular system.

[0216] A common lung disease known as chronic obstructive pulmonary disease (COPD) is often accompanied by emphysema or chronic bronchitis. Common symptoms include coughing, mucus buildup, exhaustion, wheezing, and respiratory infections.

[0217] Lung diseases such as chronic bronchitis and emphysema constrict the airways. Consequently, there is a restriction in airflow to and from the lungs, leading to dyspnea or shortness of breath. When performing pulmonary function tests, COPD is identified by its typical low airflow.

[0218] Chronic bronchitis is caused by inflammation of the major airways and lung damage. A characteristic feature of chronic bronchitis in the airways of the lungs is an increase in the volume (hyperplasia) and size (hypertrophy) of airway goblet cells and mucous glands. Consequently, there is more mucus than usual in the airways, which contributes to airway narrowing and leads to a cough that produces phlegm. Under a microscope, the walls of the airways are infiltrated by inflammatory cells. Remodeling and scarring occur after inflammation, which thickens the walls and further restricts the airways. Squamous metaplasia (abnormal changes in the tissue lining the inside of the airways) and fibrosis (additional thickening and scarring of the airway walls) occur when chronic bronchitis worsens. These changes have the effect of restricting airflow and making breathing difficult.

[0219] When someone develops asthma, a chronic lung disease, the airways become inflamed and constrict. Chronic wheezing, chest tightness, shortness of breath, and coughing are symptoms of asthma. Edema and excessive mucus production can further narrow the airways and worsen symptoms. According to the literature [Roberts et al. (1995) Chest 107:111 S-117S] (the entire text incorporated herein by reference), there is evidence that enhanced matrix degradation can occur in asthma and that this may contribute to mechanical changes in the airways. Asthma symptoms can be improved with extracellular matrix degradation therapy.

[0220] Cystic fibrosis, a multiline recessive genetic disorder, causes inappropriate chloride and sodium transport across the epithelium, resulting in thick, viscous secretions in the lungs, pancreas, liver, colon, and reproductive tract. Changes in the gene encoding the cystic fibrosis transmembrane conduction regulator (CFTR) protein cause the disease. Airway obstruction caused by mucus accumulation, reduced mucociliary clearance, and subsequent inflammation can lead to pulmonary fibrosis and anatomical abnormalities. Some patients with cystic fibrosis require a lung transplant because fibrotic lung damage worsens over time.

[0221] Only thick mucus accumulation, production of large amounts of sputum, frequent chest infections, cough, shortness of breath, inflammation, reduced exercise capacity, opportunistic infections of the lungs and sinuses (including, but not limited to, Staphylococcus aureus, Haemophilus influenzae, Mycobacterium avium, and Pseudomonas aeruginosa), pneumonia, tuberculosis, and bronchitis.

[0222] Other uses of compounds of formula (I) or formula (II) include the treatment or prevention of fibrotic diseases, disorders, or problems caused by the development of postoperative adhesions. Surgical complications, such as the development of postoperative adhesions, are frequent. Adhesions resulting from mechanical injury, ischemia, and infection can increase postoperative morbidity and mortality. Although skilled surgical techniques can reduce the severity of adhesion formation, adhesions are rarely removed, requiring the use of effective adjuvant therapy. Reducing fibrosis caused by this procedure can aid in healing and recovery by lowering discomfort, obstruction, and other surgical outcomes.

[0223] Mammalian tissue damage, such as lacerations and wounds, causes microvascular coagulation and tissue destruction at the wound surface. This tissue repair is an organized and well-controlled cellular response to injury. Regardless of their size, soft tissue injuries heal similarly. Cell proliferation and angiogenesis occur in the presence of an oxygen gradient in the biological systems responsible for tissue growth and repair. According to the literature [Hunt, TK, et al., "Coagulation and macrophage stimulation of angiogenesis and wound healing," in The Surgical Wound, pp. 1-18, ed. F. Dineen & G. Hildrick-Smith (Lea & Febiger, Philadelphia: 1981)], the sequential morphological and structural changes occurring during tissue repair have been characterized in detail and, in some cases, quantified. Cell morphology is divided into three distinct zones. The core avascular wound zone exhibits high lactate levels and is acidic, hypercarbonate, and oxygen-deficient. A gradient zone of local ischemia (ischemia) containing dividing fibroblasts is located next to the wound site. An area of ​​active collagen production, or neovascularization, may be found behind the leading zone. U.S. Patents No. 5,015,629 and No. 7,022,675 (each incorporated herein by reference) describe procedures and materials for accelerating wound healing.

[0224] By providing a compound of formula (I) or formula (II) to a patient requiring therapy, in some embodiments, the formation of scars in the patient may be reduced or stopped. Scarring is a typical side effect of healing. Excessive, thick, or raised scars can occur in wounds as a result of disordered collagen production and deposition. Generally, larger wounds take longer to heal and are more likely to leave bothersome scars.

[0225] In other embodiments, a compound of formula (I) or formula (II) may be used to reduce or stop the development of skin or sclerodermal scars. Scars on the skin can take various forms. A raised, pinkish-red area within the boundaries of the initial injury is known as a hypertrophic scar. Hypertrophic scars may sometimes fade and shrink naturally. A raised, intense red spot known as a keloid typically covers an area larger than the original injury. Keloids typically revert even after surgical removal. Skin depressions known as atrophic scars can sometimes occur due to severe acne. These are triggered by inflammation, which breaks down collagen during reconstruction and leaves a sunken area.

[0226] Compounds of formula (I) or formula (II) may be used in some embodiments to treat or prevent systemic sclerosis. Systemic connective tissue disease known as systemic sclerosis is characterized by changes in microvascular structure, alterations in the immune system, and significant accumulation of collagen and other matrix materials in connective tissues. Connective tissues of the skin and internal organs such as the gastrointestinal tract, lungs, heart, and kidneys are affected by systemic sclerosis, a clinically heterogeneous general pathological condition. Reducing fibrosis caused by systemic sclerosis can reduce symptoms and / or stop further problems in the affected tissues.

[0227] In other embodiments, a compound of formula (I) or formula (II) may be used to treat or prevent liver fibrosis. Liver fibrosis may result from chronic liver disease, virus-induced cirrhosis, hepatitis B virus infection, hepatitis C virus infection, hepatitis D virus infection, schistosomiasis, primary biliary cirrhosis, alcoholic liver disease or non-alcoholic steatohepatitis (NASH), NASH-related cirrhosis, obesity, diabetes, protein malnutrition, coronary artery disease, autoimmune hepatitis, cystic fibrosis, alpha-1-antitrypsin deficiency, primary biliary cirrhosis, drug reactions, and exposure to toxins.

[0228] NASH is a common liver disease. It is similar to alcoholic liver disease but occurs in people who drink little to no alcohol. Liver fat, along with inflammation and damage, is a major feature of NASH. However, NASH can progress to cirrhosis, a condition in which the liver is permanently scarred and damaged, rendering it unable to function normally.

[0229] NASH is generally an asymptomatic disease with few or no symptoms. Patients typically do not experience symptoms in the early stages of the disease and only begin to experience symptoms such as weakness, fatigue, and weight loss when the disease progresses or cirrhosis develops. NASH can develop over several years or even decades. Without therapy, the course may stop, and in some cases, it may even begin to reverse on its own. In some cases, NASH gradually worsens, leading to the development of liver fibrosis or scarring. As fibrosis progresses, cirrhosis appears, causing the liver to scar, harden, and become unable to perform normal functions. While cirrhosis does not always occur in NASH patients, once significant scarring or cirrhosis is present, there are few medications available to stop its progression. Patients with cirrhosis experience fluid retention, muscle atrophy, gastrointestinal bleeding, and liver failure. Progressive cirrhosis accompanied by liver failure can only be treated by liver transplantation, which is being performed increasingly frequently in NASH patients.

[0230] Compounds of formula (I) or formula (II) may be used to treat or prevent renal fibrosis in some embodiments. Dialysis after renal failure, catheter insertion, nephropathy, glomerulosclerosis, glomerulonephritis, chronic renal failure, acute renal injury, end-stage renal disease, or renal failure can all cause renal fibrosis.

[0231] Renal fibrosis is caused by the excessive accumulation of fibrous connective tissue. As a result of renal fibrosis, which has a major impact on morbidity and mortality, dialysis or a kidney transplant may be required. The filtration or reabsorption portions of the nephron, the functional unit of the kidney, can cause fibrosis. Scarring in the kidney can be caused by various factors, most notably physiological disorders affecting glomerular filtration autoregulation. This subsequently leads to the accumulation of extracellular matrix, which replaces normal structures. Numerous peptide and non-peptide fibrin sources are produced in individual cells as a result of various physiological changes, which alter the ratio of extracellular matrix synthesis to degradation in a manner favorable to scarring.

[0232] In some examples, inflammation may be one of the signs of tracheal fibrosis. According to these embodiments, a therapeutically effective amount of a compound of formula (I) or formula (II) provided to a subject requiring it may reduce or decrease the number of inflammatory cells in a tissue or organ. To determine whether the number of inflammatory cells has decreased or increased, an appropriate sample may be taken from the subject. In a non-limiting embodiment, a decrease in the number of neutrophils in bronchoalveolar lavage (BAL) fluid from a cystic fibrosis (CF) subject may be used to demonstrate a positive effect. A primary therapeutic target for CF patients is the excessive mobilization of neutrophils into their airways, which is a strong predictor of the severity of their lung disease.

[0233] In another embodiment, a reduction of one or more inflammatory biomarkers in a suitable sample from a subject may be used to measure the positive effect of a compound of formula (I) or formula (II). Inflammatory biomarkers may include one or more cytokines associated with fibrosis or inflammatory cytokines in various non-limiting embodiments. In BAL fluid, these cytokines may include, for example, IL1, MIP2 (e.g., CCL3 or CCL4), IFN, TGF, TNF, IL-6, MCP-1, IL2, and IL-10. ELISA is just one of many widely used techniques to determine the concentrations of these biomarkers. Thus, in one example, the procedure may further include lowering the levels of one or more inflammatory biomarkers in a subject sample compared to a control.

[0234] In other examples, a method for lowering or reducing collagen secretion or collagen deposition in tissues or organs such as the lungs, liver, skin, or heart of a subject may use a compound of Formula (I) or Formula (II). The procedure may involve providing a clinically effective dose of a compound of Formula (I) or Formula (II) to a patient requiring this. A person may have excessive collagen secretion or deposition in tissues or organs such as the kidneys, lungs, liver, intestines, colon, skin, or heart, or may be at risk of such pathological conditions. Generally, injury or damage is the cause of excessive collagen secretion or deposition in organs. These damage and harmful factors target specific organs. The amount of collagen deposition in tissues or organs may be reduced or lowered, either wholly or partially, by using a compound of Formula (I) or Formula (II) over a sufficiently long period of time. One week, one week to one month, one month to two months, or even more than two months may be considered a sufficient amount of time. Compounds of formula (I) or formula (II) can be used for the remainder of the subject's life when treating chronic conditions.

[0235] Atopic dermatitis

[0236] In some embodiments, a method for treating eczema in a subject requiring treatment for eczema is provided, comprising the step of administering a therapeutically effective amount of a compound of formula (I) or formula (II), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of a compound of formula (I) or formula (II).

[0237] In some variations, eczema is atopic dermatitis. Atopic dermatitis is a condition that causes redness and itching of the skin of the affected subject. Atopic dermatitis can be chronic and may recur periodically. Symptoms of atopic dermatitis may include, for example, dry skin; itching; red to brownish-gray spots; small, raised bumps that may cause fluid and / or crusting when scratched; scaly skin; and skin sensitive to scratching. Subjects with atopic dermatitis may also experience other conditions such as asthma or hay fever. Atopic dermatitis can affect children, adolescents, and adults.

[0238] In some variations, the composition of the present invention is used to treat eczema in humans and other animals with naturally occurring eczema, for example, including atopic dermatitis.

[0239] In some embodiments, the composition administered is a personal care product. In one variation, the composition administered is a cosmetic, a finished pharmaceutical product, or both. In certain variations, the composition administered meets U.S. Food and Drug Administration regulations that include requirements defining a cosmetic, a finished pharmaceutical product, or both.

[0240] In some variations, the method provided herein alleviates, slows down, or stops the development or progression of one or more symptoms caused by or associated with eczema, including, for example, atopic dermatitis. In other variations, the method provided herein reduces the severity of eczema or alleviates eczema by causing an improvement in one or more symptoms.

[0241] In some embodiments, “therapeutic effective dose” refers to an amount effective for treating a disease when administered to a subject, for example, the therapeutic effective dose may be an amount sufficient to treat eczema, for example, including atopic dermatitis. The therapeutic effective dose may be experimentally verified.

[0242] In some embodiments, the composition is administered to at least a part of the body where the subject may suffer from eczema or one or more symptoms of eczema. In some embodiments, the composition is administered to at least a part of the subject's hands, arms, legs, face and neck, and torso where the subject may suffer from eczema or one or more symptoms of eczema. In another embodiment, the composition is administered to at least a part of the entire body where the subject may suffer from eczema or one or more symptoms of eczema. In yet another embodiment, the composition is administered to at least a part of the entire body where the subject is expected to suffer from eczema or may be expected to have one or more symptoms of eczema but does not yet suffer from eczema or one or more symptoms of eczema.

[0243] In some variations, the composition is administered to a subject having mild to severe eczema, for example, including atopic dermatitis. In some variations, the composition is administered to a subject having mild to moderate eczema, for example, including atopic dermatitis. In certain variations, about 5% to about 20% of the subject's total body surface area, excluding the face, scalp, armpits, and interstitial areas, is affected by eczema, for example, including atopic dermatitis.

[0244] In some embodiments, administering the composition to a subject reduces redness, average lesion size, itching, or pain caused by eczema (e.g., including atopic dermatitis), or improves the general appearance of the affected area by eczema, or any combination of the above.

[0245] In one embodiment, a method for reducing redness in a subject suffering from eczema is provided, comprising the step of administering to a subject any composition comprising a compound of formula (I) or formula (II) as described herein or a pharmaceutical composition. In one variation of the above embodiment, the method reduces redness in a subject suffering from atopic dermatitis. In some variations, redness is evaluated on a scale of 0 to 10, where 0 corresponds to no redness and 10 corresponds to the worst conceivable redness. In some embodiments, the method comprises the step of reducing redness by about 10, 9, 8, 7, 6, 5, about 4, about 3, about 2, or about 1 point from the baseline redness of the subject evaluated before administration of the composition. In some embodiments, the method comprises the step of reducing redness by about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, or about 10% from the baseline redness of the subject evaluated before administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce redness by about 10 points, 9 points, 8 points, 7 points, 6 points, 5 points, about 4 points, about 3 points, about 2 points, or about 1 point from the baseline redness of the subject evaluated before administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce redness by about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, or about 10% from the baseline redness of the subject evaluated before administration of the composition.

[0246] In one embodiment, a method for reducing the severity of eczema in a subject suffering from eczema is provided, comprising the step of administering to a subject any composition comprising a compound of formula (I) or formula (II) as described herein or a pharmaceutical composition. In one variation of the above-described embodiment, the method reduces the severity of atopic dermatitis in a subject suffering from atopic dermatitis. In some embodiments, severity is evaluated on a scale of 0 to 4 points, where 0 corresponds to none and 4 corresponds to severe. In some embodiments, severity is evaluated based on one or more criteria selected from the group consisting of erythema, induration or papules, and oozing or crusts. In some embodiments, severity is evaluated on an investigator's comprehensive rating (IGA) scale of 0 to 4 points as described in detail in Table 5 of this invention. In some embodiments, severity is evaluated on a participant's comprehensive rating (PGA) scale of 0 to 4 points. In some embodiments described above, the severity of eczema is evaluated on an Eczema Area and Severity Index (EASI) score of 0 to 72. In some embodiments, the method comprises the step of reducing the severity on the IGA scale by about 4, about 3, about 2, or about 1 point from the baseline severity of the subject evaluated before administration of the composition. In some embodiments, the method comprises the step of reducing the severity on the PGA scale by about 4, about 3, about 2, or about 1 point from the baseline severity of the subject evaluated before administration of the composition. In some embodiments, the method comprises the step of reducing the severity on the EASI scale by about 72, about 64, about 56, about 48, about 40, about 32, about 24, about 26, or about 8 points from the baseline severity of the subject evaluated before administration of the composition.In some embodiments, the method comprises the step of reducing the severity by about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, or about 10% from the baseline severity of the subject assessed before administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce the severity on the IGA scale by about 4 points, about 3 points, about 2 points, or about 1 point from the baseline severity of the subject assessed before administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce the severity on the PGA scale by about 4 points, about 3 points, about 2 points, or about 1 point from the baseline severity of the subject assessed before administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce the severity on the EASI scale from the baseline severity of the subject assessed before administration of the composition by about 72, about 64, about 56, about 48, about 40, about 32, about 24, about 26, or about 8 points. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce the severity from the baseline severity of the subject assessed before administration of the composition by about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, or about 10%.

[0247] In one embodiment, a method for improving the appearance of the skin of a subject suffering from eczema is provided, comprising the step of administering to a subject any composition comprising a compound of formula (I) or formula (II) as described herein or a pharmaceutical composition. In one variation of the above-described embodiment, the method improves the appearance of the skin of a subject suffering from atopic dermatitis. In some embodiments, the improvement in the appearance of the skin is evaluated by the subject suffering from atopic dermatitis. In some embodiments, the improvement in the appearance of the skin is measured by a 1 to 7 point Composite Change Impression (GIC) scale. In some embodiments, the method results in skin that is significantly improved, significantly improved, minimally improved, or unchanged in appearance compared to the appearance of the skin immediately before administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to result in skin that is significantly improved, significantly improved, minimally improved, or unchanged in appearance compared to the appearance of the skin immediately before administration of the composition.

[0248] In one embodiment, a method for reducing the severity of a target lesion in a subject suffering from eczema is provided, comprising the step of administering to a subject any composition comprising a compound of formula (I) or formula (II) as described herein or a pharmaceutical composition. In one variation of the aforementioned embodiment, the method reduces the severity of a target lesion in a subject suffering from atopic dermatitis. In some embodiments, the severity of the target lesion is evaluated based on one or more criteria selected from the group consisting of erythema, edema or papules, oozing or crusting, abrasions, lichenification, and dryness. In some embodiments, the target lesion is at least 10 cm 2 It has an area of

[0249] In one embodiment, a method for reducing the average lesion size in a subject suffering from eczema is provided, comprising the step of administering to a subject any composition comprising a compound of formula (I) or formula (II) as described herein or a pharmaceutical composition. In one variation of the above-described embodiment, the method reduces the average lesion size in a subject suffering from atopic dermatitis. In some embodiments, the method comprises the step of reducing the average lesion size by at least about 100%, at least about 90%, at least about 80%, at least about 70%, at least about 60%, at least about 50%, at least about 40%, at least about 30%, at least about 20%, or at least about 10% from the baseline average lesion size of the subject as measured before administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce the average lesion size of the subject by at least about 100%, at least about 90%, at least about 80%, at least about 70%, at least about 60%, at least about 50%, at least about 40%, at least about 30%, at least about 20%, or at least about 10% from the baseline average lesion size of the subject as measured before administration of the composition.

[0250] In one embodiment, a method for reducing itching in a subject suffering from eczema is provided, comprising the step of administering to a subject any composition comprising a compound of formula (I) or formula (II) as described herein or a pharmaceutical composition. In one variation of the aforementioned embodiment, the method reduces itching in a subject suffering from atopic dermatitis. In some embodiments, itching is evaluated on a Likert scale of 0 to 10, where 0 corresponds to no itching and 10 corresponds to the worst conceivable itching. In some embodiments, itching is evaluated based on the subject's experience over the past 24 hours. In some embodiments, itching is evaluated on 7, 14, 21, or 28 days after the start of administration of the composition. In some embodiments, the method comprises the step of reducing itching assessed by about 10 points, about 9 points, about 8 points, about 7 points, about 6 points, about 5 points, about 4 points, about 3 points, about 2 points, or about 1 point from the itching baseline assessment of a subject prior to administration of the composition. In some embodiments, the method comprises the step of reducing itching by about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, or about 10% from the itching baseline assessment of a subject prior to administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce itching assessed by about 10 points, about 9 points, about 8 points, about 7 points, about 6 points, about 5 points, about 4 points, about 3 points, about 2 points, or about 1 point from the itching baseline assessment of the subject prior to administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce itching by about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, or about 10% from the itching baseline assessment of the subject prior to administration of the composition.

[0251] In one embodiment, a method for reducing pain in a subject suffering from eczema is provided, comprising the step of administering to a subject any composition comprising a compound of formula (I) or formula (II) as described herein or a pharmaceutical composition. In one variation of the aforementioned embodiment, the method reduces pain in a subject suffering from atopic dermatitis. In some embodiments, pain is evaluated on a Likert scale of 0 to 10, where 0 corresponds to no pain and 10 corresponds to the worst imaginable pain. In some embodiments, pain is evaluated based on the subject's experience over the past 24 hours. In some embodiments, pain is evaluated on days 7, 14, 21, or 28 after the start of administration of the composition. In some embodiments, the method comprises the step of reducing pain assessed by about 10 points, about 9 points, about 8 points, about 7 points, about 6 points, about 5 points, about 4 points, about 3 points, about 2 points, or about 1 point from a baseline assessment of pain in a subject prior to administration of the composition. In some embodiments, the method comprises the step of reducing pain assessed by about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, or about 10% from a baseline assessment of pain in a subject prior to administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce pain assessed by about 10 points, about 9 points, about 8 points, about 7 points, about 6 points, about 5 points, about 4 points, about 3 points, about 2 points, or about 1 point from the baseline pain assessment in a subject prior to administration of the composition. In some embodiments, the method comprises the step of administering the composition for a period sufficient to reduce pain by about 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, or about 10% from the baseline pain assessment in a subject prior to administration of the composition.

[0252] In some variations of the above description, the subject is formally or clinically diagnosed with eczema, for example, including atopic dermatitis.

[0253] manufacturing of medicine

[0254] A method for preparing a compound such as the compound of formula (Iaa) comprises the steps of: providing a solution of isoandrographolide and a solvent; forming a first intermediate by combining 2,2,6,6-tetramethylpiperidinyloxy (TEMPO), tetrabutylammonium iodide (TBAI), N-chlorosuccinimide (NCS), a salt, and a solution; forming a second intermediate by combining hydroxylamine hydrochloride, pyridine, and the first intermediate; and dehydrating the second intermediate with cyanide to form the compound of formula (Iaa).

[0255] In some embodiments, the dehydration step with cyanide comprises combining triethylamine, 4-toluenesulfonyl chloride, and a second intermediate to form a compound of formula (Iaa). In other embodiments, the dehydration step with cyanide comprises combining benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), 1,8-diazabicyclo[5.4.0]undec-7-en (DBU), and a second intermediate to form a compound of formula (Iaa).

[0256] In a specific embodiment, a method for preparing a compound such as the compound of formula (IIaa) comprises the steps of: providing a solution of isoandrogramolide and a solvent; combining tert-butyldimethylsilyl chloride (TBDMSCl) and the solution to form a first intermediate; combining pyridine, celite, pyridinium chlorochromate and the first intermediate to form a second intermediate; combining hydroxylamine hydrochloride, pyridine and the second intermediate to form a third intermediate; and combining 4-toluenesulfonyl chloride and the third intermediate and then treating with dioxane HCl to form the compound of formula (IIaa).

[0257] In another embodiment, a method for preparing a compound such as the compound of formula (IIaa) comprises the steps of: providing a solution of isoandrographolide and a solvent; combining dess-martin periodinane (DMP) and the solution to form a first intermediate; combining hydroxylamine hydrochloride, pyridine, and the first intermediate to form a second intermediate; and combining triethylamine, 4-methylbenzene-1-sulfonyl chloride, 4-dimethylaminopyridine (DMAP), and the second intermediate, and then treating with dioxane HCl to form the compound of formula (IIaa).

[0258] In another embodiment, a method for preparing a compound such as compound 3 as described below is illustrated in FIG. 17.

[0259] Administration of pharmaceutically acceptable salts and compositions

[0260] In specific embodiments of the present disclosure, the “effective amount” of a compound, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable composition is an amount effective for treating or reducing the severity of one or more of chronic pain, intestinal pain, neuropathic pain, musculoskeletal pain, acute pain, inflammatory pain, cancer pain, idiopathic pain, pruritus, atopic dermatitis, or psoriasis.

[0261] Compounds and compositions of Formula (I) or Formula (II) according to the method of the present disclosure may be administered in any amount and via any route of administration effective to treat or reduce the severity of one or more of the painful or non-painful conditions mentioned herein. The exact amount required will vary by subject depending on the species, age, and general condition of the subject, the severity of the infection, the specific agent, the mode of administration, etc. Compounds of Formula (I) or Formula (II) of the present disclosure may be formulated in a dosage unit form for ease of administration and dosage uniformity. The expression “supply unit form” refers to a physically distinct unit of the preparation suitable for the subject to be treated when used herein. However, it will be understood that the total daily dose of compounds and compositions of Formula (I) or Formula (II) of the present disclosure will be determined by the attending physician within the scope of appropriate medical judgment. The specific effective dose level for any specific subject or organism is determined by the disorder being treated and the severity of the disorder; the activity of the specific compound used; the specific composition used; It will depend on age, weight, the subject's general health, gender, and dietary habits; the time of administration, the route of administration, and the elimination rate of the specific compound used; the duration of treatment; various factors including drugs used in combination or simultaneously with the specific compound used, and similar factors known in the medical field. As used herein, the terms "subject" or "patient" mean animals, preferably mammals, and most preferably humans.

[0262] The pharmaceutically acceptable compositions of the present disclosure may be administered to humans and other animals orally, rectally, parenterally, intravaginally, vaginally, intraperitoneally, topically (as a powder, ointment, or mucous), or buccally as an oral or nasal spray, depending on the severity of the infection being treated. As used herein, the terms “parenteral administration” and “parenterally administered” mean a mode of administration other than enteric and topical administration by ordinary injection, and include, but are not limited to, intravenous, intramuscular, intra-arterial, intravertebral, intrasacral, intraorbital, intracardiac, intradermal, intraperitoneal, transtubercular, subcutaneous, subcutaneous, intra-articular, subcapsular, subarachnoid, intravertebral, and intrasternal injections and infusions. In certain embodiments, the compound of formula (I) or formula (II) of the present disclosure may be administered orally or parenterally at dose levels of about 0.01 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 50 mg / kg, and optionally about 1 mg / kg to about 25 mg / kg per day, at least once a day to obtain the desired therapeutic effect.

[0263] Liquid formulations for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, the liquid formulation may contain inert diluents commonly used in the art, e.g., water or other solvents, solubilizers and emulsifiers, e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (specifically cottonseed oil, peanut oil, corn oil, sprout oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, sorbitan fatty acid esters, and mixtures thereof. In addition to inert diluents, the oral composition may also include adjuvants, such as humectants, emulsifiers, suspending agents, sweeteners, flavoring agents, and fragrances.

[0264] In addition to the active compound, the suspension may contain, for example, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0265] It is known that sterols, such as cholesterol, form complexes with cyclodextrins. Therefore, in certain embodiments, when the inhibitor is a steroidal alkaloid, it may be formulated with cyclodextrins, such as α-, β-, and γ-cyclodextrins, dimethyl-β-cyclodextrin, and 2-hydroxypropyl-β-cyclodextrin.

[0266] Injectable formulations, e.g., sterile aqueous injectable solutions or oily suspensions, may be formulated according to techniques known in the art using suitable dispersants or wetting agents and suspending agents. Sterile injectable formulations may also be sterile injectable solutions, suspensions, or emulsions in diluents or solvents acceptable for non-toxic parenteral administration, e.g., solutions in 1,3-butanediol. Among the acceptable excipients and solvents that may be used are water, Ringer's solution, USP, and isotonic sodium chloride solutions. Additionally, sterile non-volatile oils are commonly used as solvents or suspension media. For this purpose, non-irritating bland fixed oils containing synthetic monoglycerides or diglycerides may be used. Furthermore, fatty acids such as oleic acid are used to manufacture injectable formulations.

[0267] The injectable formulation may be sterilized, for example, by filtration through a bacterial capture filter, by autoclave sterilization, or by incorporating a sterile agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable media before use. In various embodiments, a first portion of the formulation may be sterilized by filtration (e.g., a compound), and a second portion may be sterilized by autoclave sterilization (e.g., an excipient).

[0268] To prolong the effects of the compounds of the present disclosure, it is often desirable to slow the absorption of the compounds from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material that is poorly soluble. The absorption rate of the compound depends subsequently on its dissolution rate and, consequently, may depend on crystal size and crystal form. Alternatively, delayed absorption of parenterally administered compound forms is achieved by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are prepared by forming a microcapsule substrate of the compound in a biodegradable polymer such as polylactide-polyglycolide. The release rate of the compound can be controlled depending on the ratio of the compound to the polymer and the properties of the specific polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations can also be prepared by encapsulating the compound in liposomes or microemulsions that are compatible with biological tissues.

[0269] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound comprises at least one inert pharmaceutically acceptable excipient or carrier, e.g., sodium citrate, dicalcium phosphate and / or a) fillers or extenders, e.g., starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders, e.g., carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; c) humectants, e.g., glycerol; d) disintegrants, e.g., agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) dissolution retardants, e.g., paraffin; f) an absorption promoter, e.g., a quaternary ammonium compound, g) a wetting agent, e.g., cetyl alcohol and glycerol monostearate, h) an absorbent, e.g., kaolin and bentonite clay, and i) a lubricant, e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also include a buffer.

[0270] A similar type of solid composition may also be used as a filler within soft or hard-filled gelatin capsules using excipients such as lactose or lactose and high molecular weight polyethylene glycol, etc. Solid dosage forms such as tablets, coated tablets, capsules, pills, and granules may be manufactured with coatings and shells, such as enteric coatings and other coatings known in the field of pharmaceutical formulation. These may optionally contain an opacifying agent and may also be compositions that selectively release the active compound(s) alone or, preferentially, in a delayed manner in some part of the intestinal tract. Examples of embedding compositions that may be used include polymerizable materials and waxes. A similar type of solid composition may also be used as a filler within soft or hard-filled gelatin capsules using excipients such as lactose or lactose and high molecular weight polyethylene glycol, etc.

[0271] Tablets may be manufactured by compression or molding, optionally with one or more auxiliary components. Compressed tablets may be manufactured using a binder (e.g., gelatin or hydroxypropylmethylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (e.g., sodium glycolate starch or cross-linked sodium carboxymethylcellulose), a surfactant, or a dispersant. Molded tablets may be manufactured by molding a mixture of powdered compounds moistened with an inert liquid diluent in a suitable machine.

[0272] Tablets and other solid dosage forms of the pharmaceutical compositions of the present invention, such as coated tablets, capsules, pills, and granules, may optionally be scored or manufactured with coatings and shells such as enteric coatings and other coatings well known in the field of pharmaceutical formulation. Additionally, they may be formulated to provide sustained-release or controlled-release of the active ingredient using varying proportions of hydroxypropylmethylcellulose to provide desired release profiles, different polymer matrices, liposomes, and / or microspheres. They may be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating a sterile agent in the form of a sterile solid composition that can be dissolved in sterile water or some other sterile injectable medium immediately before use. These compositions may also optionally contain an opacifying agent and may have compositions that release only the active ingredient(s), or preferentially in a specific part of the gastrointestinal tract, optionally in a delayed manner. Examples of embedding compositions that may be used include polymerizable materials and waxes. The active ingredient may also be in a microencapsulated form having one or more of the excipients described above, if appropriate.

[0273] Formulations of the pharmaceutical composition of the present disclosure for rectal, vaginal, or urethral administration may be prepared by mixing one or more compounds with one or more suitable non-irritating excipients or carriers, for example, cocoa butter, polyethylene glycol, suppository wax, or salicylate, and may be provided as suppositories that are solid at room temperature but liquid at body temperature, and thus dissolve in the rectal or vaginal cavity to release the active compound.

[0274] Alternatively or additionally, the composition may be formulated for delivery via a catheter, stent, wire, or other intraductal device. Delivery via such devices may be particularly useful for delivery to the bladder, urethra, ureter, rectum, or small intestine.

[0275] Formulations suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams, or spray formulations containing a carrier as known in the art to be suitable.

[0276] The active compound may also be in a micro-encapsulated form with one or more of the excipients mentioned above. Solid dosage forms such as tablets, coated tablets, capsules, pills, and granules may be manufactured using coatings and sheaths, such as enteric coatings, controlled-release coatings, and other coatings well known in the field of pharmaceutical formulations. In such solid dosage forms, the active compound may be mixed with at least one inert diluent, such as sucrose, lactose, or starch. Such dosage forms may also, as in conventional practice, include additional materials other than the inert diluent, such as tablet lubricants and other tablet aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms may also include a buffer. These may optionally contain an opacifying agent and may also be compositions that release only the active ingredient(s) in a delayed manner, preferentially at specific sites within the intestinal tract. Examples of embedding compositions that may be used include polymerizable materials and waxes.

[0277] Dosage forms for topical or transdermal administration of the compounds of the present disclosure include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compounds may be mixed under sterile conditions with a pharmaceutically acceptable carrier and with any preservatives, buffers, or propellants that may be required.

[0278] Dosage forms for topical or transdermal administration of the compound include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier and with any preservatives, buffers, or propellants that may be required.

[0279] Ointments, pastes, creams, and gels, in addition to the active compound of the present invention, comprise excipients, such as animal and vegetable fats, oils, waxes, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, and zinc oxide, or mixtures thereof.

[0280] In addition to the compounds of the present invention, the powder and spray may contain excipients, such as lactose, talc, silica, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these materials. The spray may additionally contain conventional propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0281] Transdermal patches have the additional advantage of providing controlled delivery of compounds into the body. Such dosage forms can be prepared by dissolving or dispersing the compound in a suitable medium. Absorption promoters may also be used to increase the amount of compound penetrating the skin. The rate of this flow can be controlled by providing a rate control membrane or by dispersing the compound in a polymer matrix or gel.

[0282] Ophthalmic formulations, ophthalmic ointments, eye drops, and ear drops are also considered within the scope of this disclosure. A pharmaceutical composition suitable for parenteral administration comprises one or more compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions immediately before use, which may contain antioxidants, buffers, bacteriostatic agents, solutes that make the formulation isotonic with the blood of the intended recipient, or suspenders or thickeners.

[0283] Examples of suitable aqueous and non-aqueous carriers that can be used in pharmaceutical compositions include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils, e.g., olive oil, and injectable organic esters, e.g., ethyl oleate. Suitable fluidity can be maintained, for example, by the use of a coating material such as lecithin, by maintaining the particle size required in the case of a dispersion, and by the use of a surfactant.

[0284] These compositions may also contain auxiliary agents such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of microbial action can be ensured by the inclusion of various antimicrobial and antifungal agents, e.g., parabens, chlorobutanol, phenol sorbic acid, etc. It may also be desirable to include isotonic agents in the composition, e.g., sugars, sodium chloride, etc. Additionally, extended absorption in the injectable pharmaceutical form may occur by the incorporation of agents that delay absorption, such as aluminum monostearate and gelatin.

[0285] In some cases, to prolong the effect of the drug, it is desirable to slow down the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of a crystalline or amorphous material having poor water solubility. In this case, the rate of drug absorption depends on its dissolution rate, and thus on crystal size and crystalline form. Alternatively, delaying the absorption of parenteral drug forms is achieved by dissolving or suspending the drug in an oily excipient.

[0286] When a compound of formula (I) or formula (II) is administered to humans and animals as a pharmaceutical agent, it may be provided as a pharmaceutical composition containing 0.1 to 99.5% (more preferably 0.5 to 90%) of an active ingredient, either by itself or in combination with, for example, a pharmaceutically acceptable carrier.

[0287] Adding an active compound to animal feed is preferably achieved by preparing a suitable feed premix containing an effective amount of the active compound and incorporating the premix into the complete feed.

[0288] Alternatively, intermediate concentrates or feed supplements containing active ingredients may be blended into the feed. Methods for preparing and administering such feed premixes and complete feeds are described in reference books (e.g., literature ["Applied Animal Nutrition", WH Freedman and CO., San Francisco, USA, 1969 or "Livestock Feeds and Feeding" O and B books, Corvallis, Ore., USA, 1977]).

[0289] The delivery method may also be provided by rechargeable or biodegradable devices. Various sustained-release polymerizable devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including protein biopharmaceuticals. Various biocompatible polymers (including hydrogels), comprising both biodegradable and non-degradable polymers, can be used to form sustained-release implants of compounds at specific target sites.

[0290] The actual dosage level of the active ingredient in the pharmaceutical composition can be varied to obtain an amount of the active ingredient effective in achieving a desired therapeutic response for a specific patient, composition, and mode of administration without toxicity to the patient.

[0291] The selected dosage level will be determined by various factors including the activity of the specific compound, or its ester or salt or amide, used, the route of administration, the time of administration, the excretion rate of the specific compound being used, the duration of treatment, other drugs, compounds, and / or substances used in combination with the specific compound being used, the age, gender, weight, condition, general health, and previous medical history of the patient being treated, and other factors sufficiently known in the medical field.

[0292] A physician or veterinarian skilled in the art can easily determine and prescribe the effective amount of the necessary pharmaceutical composition. For example, a physician or veterinarian could start administering the compound of formula (I) or formula (II) used in the pharmaceutical composition at a level lower than necessary to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved.

[0293] Generally, the appropriate daily dose of the compound will be the amount of the compound that is the lowest effective dose to produce a therapeutic effect. This effective dose will generally depend on the factors described above. Generally, the intravenous, intraventricular, and subcutaneous doses of the compound of formula (I) or formula (II) for a patient will be in the range of about 0.0001 to about 100 mg per kilogram of body weight per day.

[0294] If desired, the effective daily dose of the active compound may be administered, optionally in unit dose form, divided into 2, 3, 4, 5, 6 or more sub-dose doses at appropriate intervals throughout the day.

[0295] Patients receiving such treatment include any animals as needed, e.g., primates, particularly humans and other mammals, e.g., horses, cattle, pigs, and sheep; and generally poultry and pets.

[0296] The compound may be administered in this manner, or as a mixture with a pharmaceutically acceptable and / or sterile carrier, and may also be administered with other antimicrobial agents such as penicillin, cephalosporin, aminoglycosides, and glycopeptides. Thus, combination therapy involves sequential, simultaneous, and separate administration of the active compound in such a manner that the therapeutic effects of the first administration are still detectable when the subsequent therapy is administered.

[0297] The present invention considers the formulation of the compound in any of the aforementioned pharmaceutical compositions and formulations. Additionally, the present invention considers administration via any of the aforementioned routes of administration. Those skilled in the art may select an appropriate formulation and route of administration based on the condition being treated and the overall health, age, and size of the patient being treated.

[0298] The activity of the compounds used in this disclosure as inhibitors of therapeutic targets can be tested according to the methods generally described in the embodiments of this invention, or according to methods available to those skilled in the art.

[0299] Additional treatment

[0300] Furthermore, it will be understood that the compounds of Formula (I) or Formula (II) and pharmaceutically acceptable compositions of the present disclosure may be used in combination therapy, that is, the compounds of Formula (I) or Formula (II) and pharmaceutically acceptable compositions may be administered simultaneously, prior to, or after one or more other desired therapeutic agents or medical procedures. The specific combination of therapies (treatments or courses) to be used in combination therapy will take into account the compatibility of the desired treatment and / or course and the desired therapeutic effect to be achieved. Additionally, it will be understood that the therapies used may achieve the desired effect for the same disorder (e.g., the compounds of the present invention may be administered simultaneously with other agents used to treat the same disorder) or achieve different effects (e.g., control of any side effects). Additional therapeutic agents orally administered to treat or prevent a specific disease or condition used herein are known as "suitable for the disease or condition being treated." For example, exemplary additional therapeutic agents include, but are not limited to, ursodeoxycholic acid (UDCA), rifampicin, cholestyramine, naltrexone, sertraline, phenobarbital, gabapentin, pregabalin, and ondansetron. Additionally, non-drug analgesic approaches may be used in conjunction with the administration of one or more of the compounds of this disclosure. For example, anesthetic (intrathecal injection, nerve block), neurosurgical (neurolysis of CNS pathways), neurostimulatory (transcutaneous electrical nerve stimulation, dorsal column stimulation), physical medicine (physical therapy, corrective devices, diathermy), or psychological (cognitive methods—hypnosis, biofeedback, or behavioral methods) approaches may also be used. Additional suitable therapeutic agents or approaches are generally [Merck Manual, Nineteenth Edition, Ed. Robert S. Porter and Justin L. Kaplan, Merck Sharp & Dohme Corp., the entire contents of which are incorporated herein by reference].[, a subsidiary of Merck & Co., Inc., 2011], and is listed on the Food and Drug Administration website (www.fda.gov).

[0301] In another embodiment, additional suitable therapeutic agents are selected from the following: (1) opioid analgesics, e.g., morphine, heroin, hydromorphone, oxymorphone, levorphanol, levalofane, methadone, meperidine, fentanyl, cocaine, codeine, dihydrocodeine, oxycodone, hydrocodone, propoxyphen, nalmephen, nallopin, naloxone, naltrexone, buprenorphine, butorphanol, nalbuphine or pentazocine; (2) Non-steroidal anti-inflammatory drugs (NSAIDs), e.g., aspirin, diclofenac, diflunisal, etodolac, fenbufen, fenoprofen, flufenisal, flurbiprofen, ibuprofen, indomethacin, ketoprofen, ketorolac, meclofenamic acid, mefenamic acid, meloxicam, nabumetone, naproxen, nimesulide, nitroflurbiprofen, olsalazine, oxaprozine, phenylbutazone, piroxicam, sulfasalazine, sulindac, tolmetine, or zomepirac; (3) Barbiturate sedatives, e.g., amobarbital, aprobarbital, butabarbital, butalbital, mefobarbital, metabarbital, methohexital, pentobarbital, phenobarbital, secobarbital, talbutal, thiamylal, or thiopental; (4) Benzodiazepines having a sedative effect, e.g., chlordiazepoxide, chlorazefate, diazepam, flurazepam, lorazepam, oxazepam, temazepam, or triazolam; (5) Histamine (H1) antagonists having a sedative effect, e.g., diphenhydramine, pyramine, promethazine, chlorpheniramine, or chlorcyclizine; (6) Sedatives, e.g., glutethimide, meprobamate, methaqualone, or dichlorphenazone; (7) Skeletal muscle relaxants, e.g., baclofen, carisoprodol, chlorzazone, cyclobenzaprine, methocarbamol, or orphenadine;(8) NMDA receptor antagonists, e.g., dextromethorphan ((+)-3-hydroxy-N-methylmorphinan) or its metabolite dextromethorphan ((+)-3-hydroxy-N-methylmorphinan), ketamine, memantine, pyrroloquinoline quinine, cis-4-(phosphonomethyl)-2-piperidinecarboxylic acid, budipine, EN-3231 (MorphiDex®), combination formulation of morphine and dextromethorphan), topiramate, neramexane or NR2B antagonists comprising fuzinpotel, e.g. ifenprodil, traxoprodil or (-)-(R)-6-{2-[4-(3-fluorophenyl)-4-hydroxy-1-piperidinyl]-1-hydroxyethyl-3,4-dihydro-2(1H)-quinolinone; (9) Alpha-adrenergic agents, e.g., doxazosin, tamsulosin, clonidine, guanfacine, dexmedetomidine, modafinil, or 4-amino-6,7-dimethoxy-2-(5-methane-sulfonamido-1,2,3,4-tetrahydroisonine-2-yl)-5-(2-pyridyl)quinazolin; (10) Tricyclic antidepressants, e.g., desipramine, imipramine, amitriptyline, or nortriptyline; (11) Anticonvulsants, e.g., carbamazepine (Tegretol®), lamotrigine, topiramate, lacosamide (Vimpat®), or valproate; (12) Tachykinin (NK) antagonists, particularly NK-3, NK-2, or NK-1 antagonists, e.g., (alpha-R,9R)-7-[3,5-bis(trifluoromethyl)benzyl]-8,9,10,11-tetrahydro-9-methyl-5-(4-methylphenyl)-7H-[1,4]diazosino[2,1-g][1,7]-naphthyridine-6-13-dione (TAK-637), 5-[[(2R,3S)-2-[(1R)-1-[3,5-bis(trifluoromethyl)phenyl]ethoxy-3-(4-fluorophenyl)-4-morpholinyl]-methyl]-1,2-dihydro-3H-1,2,4-triazole-3-one (MK-869), aprepitant, ranepitant, daptant, or 3-[[2-methoxy-5-(trifluoromethoxy)phenyl]-methylamino]-2-phenylpiperidine(2S,3S);(13) Muscarinic antagonists, e.g., oxybutinin, tolterodine, propiverine, tropsium chloride, darifenacin, solifenacin, temiverine, and ipratropium; (14) COX-2 selective inhibitors, e.g., celecoxib, rofecoxib, parecoxib, valdecoxib, deracoxib, etoricoxib, or lumiracoxib; (15) coal-tar analgesics, particularly paracetamol; (16) Neuroleptics, e.g. droperidol, chlorpromazine, haloperidol, perphenazine, thioridazine, mesoridazine, trifluoperazine, fluphenazine, clozapine, olanzapine, risperidone, ziprasidone, quetiapine, sertindol, aripiprazole, sonepiprazole, blonanserin, iloperidone, ferrospirone, raclopride, zotepine, bifefrunox, asenapine, lurasidone, amisulpride, valaperidone, palindor, eplibanserin, osanetant, limonabant, meclinertant, Miraxion® or sarizotan; (17) vanilloid receptor agonists (e.g., recepatoxin or cibamid) or antagonists (e.g., capsazepine, GRC-15300); (18) Beta-adrenergic agents such as propranolol; (19) Local anesthetics such as mexiletine; (20) Corticosteroids such as dexamethasone; (21) 5-HT receptor agonists or antagonists, in particular 5-HT1B / 1D agonists such as eletriptan, sumatriptan, naratriptan, zolmititan or rizatriptan; (22) 5-HT2A receptor antagonists, e.g., R(+)-alpha-(2,3-dimethoxy-phenyl)-1-[2-(4-fluorophenylethyl)]-4-piperidine methanol (MDL-100907); (23) Cholinergic (nicotine) analgesics such as ispronicline (TC-1734), (E)-N-methyl-4-(3-pyridinyl)-3-butene-1-amine (RJR-2403), (R)-5-(2-azetidinyl methoxy)-2-chloropyridine (ABT-594) or nicotine; (24) Tramadol®, Tramadol ER (Ultram ER®), tapentadol ER (Nucynta®);(25) 5-[2-ethoxy-5-(4-methyl-1-piperazinyl-sulfonyl)phenyl]-1-methyl-3-n-propyl-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one (sildenafil), (6R,12aR)-2,3,6,7,12,12a-hexahydro-2-methyl-6-(3,4-methylenedioxyphenyl)-pyrazino[2',1':6,1]-pyrido[3,4-b]indole-1,4-dione (IC-351 or tadalafil), 2-[2-ethoxy-5-(4-ethyl-piperazine-1-yl-1-sulfonyl)-phenyl]-5-methyl-7-propyl-3H-imidazo[5,1-f][1,2,4]triazine-4-one(vardenafil), 5-(5-acetyl-2-butoxy-3-pyridinyl)-3-ethyl-2-(1-ethyl-3-azetidinyl)-2,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one, 5-(5-acetyl-2-propoxy-3-pyridinyl)-3-ethyl-2-(1-isopropyl-3-azetidinyl)-2,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one, PDE5 inhibitors such as 5-[2-ethoxy-5-(4-ethylpiperazine-1-ylsulfonyl)pyridin-3-yl]-3-ethyl-2-[2-methoxyethyl]-2,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one, 4-[(3-chloro-4-methoxybenzyl)amino]-2-[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]-N-(pyrimidin-2-ylmethyl)pyrimidin-5-carboxamide, and 3-(1-methyl-7-oxo-3-propyl-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-5-yl)-N-[2-(1-methylpyrrolidin-2-yl)ethyl]-4-propoxybenzenesulfonamide;(26) Alpha-2-delta ligands, e.g., gabapentin (Neurontin®), gabapentin GR (Gralise®), gabapentin, enacarbyl (Horizant®), pregabalin (Lyrica®), 3-methyl gabapentin, (1[alpha],3[alpha],5[alpha]) (3-amino-methyl-bicyclo[3.2.0]hept-3-yl)-acetic acid, (3S,5R)-3-aminomethyl-5-methyl-heptanoic acid, (3S,5R)-3-amino-5-methyl-heptanoic acid, (3S,5R)-3-amino-5-methyl-octanoic acid, (2S,4S)-4-(3-chlorophenoxy)proline, (2S,4S)-4-(3-fluorobenzyl)-proline, [(1R,5R,6S)-6-(aminomethyl)bicyclo[3.2.0]hept-6-yl]acetic acid, 3-(1-aminomethyl-cyclohexylmethyl)-4H-[1,2,4]oxadiazole-5-one, C-[1-(1H-tetrazole-5-ylmethyl)-cycloheptyl]-methylamine, (3S,4S)-(1-aminomethyl-3,4-dimethyl-cyclopentyl)-acetic acid, (3S,5R)-3-aminomethyl-5-methyl-octanic acid, (3S,5R)-3-amino-5-methyl-nonanoic acid, (3S,5R)-3-amino-5-methyl-octanic acid, (3R,4R,5R)-3-amino-4,5-dimethyl-heptanoic acid and (3R,4R,5R)-3-amino-4,5-dimethyl-octanic acid; (27) Cannabinoids such as KHK-6188; (28) Metabolic glutamate subtype 1 receptor (mGluR1) antagonists; (29) Serotonin reuptake inhibitors, e.g., sertraline, sertraline metabolite demethylsertraline, fluoxetine, norfluoxetine (fluoxetine desmethyl metabolite), fluvoxamine, paroxetine, citalopram, citalopram metabolite desmethylcitalopram, escitalopram, d,l-fenfluramine, femoxetine, ipoxetine, cyanodotiepin, ritoxetine, dapoxetine, nefazodone, cericleamine, and trazodone;(30) Noradrenaline (norepinephrine) reuptake inhibitors, e.g., maprotiline, lopepramine, mirtazepine, oxaprotiline, pezolamine, tomoxetine, mianserine, bupropion, bupropion metabolites, hydroxybupropion, nomiphensine and viloxazine (Vivalan®), particularly selective noradrenaline reuptake inhibitors, e.g., levoxetine, particularly (S,S)-levoxetine; (31) Dual serotonin-noradrenaline reuptake inhibitors, e.g., venlafaxine, venlafaxine metabolite O-desmethylvenlafaxine, clomipramine, clomipramine metabolite desmethylclomipramine, duloxetine (Cymbalta®), milnacipran and imipramine; (32) Inducible nitric oxide synthase (iNOS) inhibitors, e.g., S-[2-[(1-iminoethyl)amino]ethyl]-L-homocysteine, S-[2-[(1-iminoethyl)amino]ethyl]-4,4-dioxo-L-cysteine, S-[2-[(1-iminoethyl)amino]ethyl]-2-methyl-L-cysteine, (2S,5Z)-2-amino-2-methyl-7-[(1-iminoethyl)amino]-5-heptene acid, 2-[[(1R,3S)-3-amino-4-hydroxy-1-(5-thiazolyl)-butyl]thio]-S-chloro-S-pyridine carbonitrile; 2-[[(1R,3S)-3-amino-4-hydroxy-1-(5-thiazolyl)butyl]thio]-4-chlorobenzonitrile, (2S,4R)-2-amino-4-[[2-chloro-5-(trifluoromethyl)phenyl]thio]-5-thiazolebutanol, 2-[[(1R,3S)-3-amino-4-hydroxy-1-(5-thiazolyl)butyl]thio]-6-(trifluoromethyl)-3-pyridinecarbonitrile, 2-[[(1R,3S)-3-amino-4-hydroxy-1-(5-thiazolyl)butyl]thio]-5-chlorobenzonitrile, N-[4-[2-(3-chlorobenzylamino)ethyl]phenyl]thiophene-2-carboxamidine, NXN-462 or Guanidinoethyl disulfide; (33) Acetylcholinesterase inhibitors such as donepezil;(34) Prostaglandin E2 subtype 4 (EP4) antagonists such as N-[({2-[4-(2-ethyl-4,6-dimethyl-1H-imidazo[4,5-c]pyridin-1-yl)phenyl]ethyl}amino)-carbonyl]-4-methylbenzenesulfonamide or 4-[(15)-1-({[5-chloro-2-(3-fluorophenoxy)pyridin-3-yl]carbonyl}amino)ethyl]benzoic acid; (35) Leukotriene B4 antagonists; e.g. 1-(3-biphenyl-4-ylmethyl-4-hydroxy-chroman-7-yl)-cyclopentanecarboxylic acid (CP-105696), 5-[2-(2-carboxyethyl)-3-[6-(4-methoxyphenyl)-5E-hexenyl]oxyphenoxy]-valeric acid (ONO-4057) or DPC-11870; (36) 5-lipoxygenase inhibitors, e.g., zilyutone, 6-[(3-fluoro-5-[4-methoxy-3,4,5,6-tetrahydro-2H-pyran-4-yl])phenoxy-methyl]-1-methyl-2-quinolone (ZD-2138), or 2,3,5-trimethyl-6-(3-pyridylmethyl)-1,4-benzoquinone (CV-6504); (37) sodium channel blockers, e.g., lidocaine, lidocaine plus tetracaine cream (ZRS-201) or esricarbazepine acetate; (38) NaV1.7 blockers, e.g., XEN-402 and those disclosed in, e.g., WO2011 / 140425; WO2012 / 106499; WO2012 / 112743; WO2012 / 125613 or PCT / US2013 / 21535 - the whole contents of each application are incorporated herein by reference -; (39) NaV1.8 blockers, e.g., those disclosed in WO2008 / 135826 and WO2006 / 011050 - the whole contents of each application are incorporated herein by reference -; (40) combined NaV1.7 and NaV1.8 blockers, e.g., DSP-2230 or BL-1021; (41) 5-HT3 antagonists, e.g., ondansetron; (42) TPRV 1 receptor agonists, e.g., capsaicin (NeurogesX®, Qutenza®); and pharmaceutically acceptable salts and solvates thereof; (43) nicotine receptor antagonists, e.g., varenicline;(44) N-type calcium channel antagonist, e.g. Z-160; (45) nerve growth factor antagonist, e.g. Tanezumab; (46) endopeptidase stimulator, e.g. Senlebotase; and (47) angiotensin II antagonist, e.g. EMA-401.;

[0302] In one embodiment, additional suitable therapeutic agents are selected from V-116517, pregabalin, controlled-release pregabalin, ezogabine (Potiga®), ketamine / amitriptyline topical cream (Amiket®), AVP-923, perampanel (E-2007), ralfinamide, transdermal bupivacaine (Eladur®), CNV1014802, JNJ-10234094 (carisbamate), BMS-954561 or ARC-4558.

[0303] The amount of additional therapeutic agent present in the composition of the present disclosure will not exceed the amount typically administered to a composition containing the therapeutic agent as a single active agent. In the composition disclosed herein, the amount of additional therapeutic agent will be in the range of about 10% to 100% of the amount typically present in a composition containing said therapeutic agent as a single therapeutic active agent.

[0304] Compounds of Formula (I) or Formula (II) of the present disclosure, or pharmaceutically acceptable compositions thereof, may also be incorporated into compositions for coating implantable medical devices, such as implants, artificial valves, vascular grafts, stents, and catheters. Accordingly, the present disclosure, in another aspect, comprises a composition for coating an implantable device comprising a compound of the present disclosure as generally described above and in the classifications and subclasses herein, and a carrier suitable for coating said implantable device. In a further aspect, the present disclosure comprises an implantable device coated with a composition comprising a compound of the present disclosure as generally described above and in the classifications and subclasses herein, and a carrier suitable for coating said implantable device. Suitable coatings and general manufactures of coated implantable devices are described in U.S. Patents No. 6,099,562; No. 5,886,026; and No. 5,304,121. The coating is typically a biocompatible polymer material, such as a hydrogel polymer, polymethyldisiloxane, polycaprolactone, polyethylene glycol, polylactic acid, ethylene vinyl acetate, and mixtures thereof. The coating may optionally be further covered by a suitable topcoat of fluorosilicone, polysaccharide, polyethylene glycol, phospholipid, or a combination thereof to impart controlled release features to the composition.

[0305] Another aspect of the present disclosure relates to inhibiting therapeutic target activity in a biological sample or subject, wherein the method comprises the step of administering to a subject or the step of contacting the biological sample with a compound of formula (I) or formula (II) or a composition comprising a compound. As used herein, the term “biological sample” includes, without limitation, cell cultures or extracts thereof; biopsy material or extracts thereof obtained from mammals; and blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.

[0306] Inhibition of therapeutic target activity in biological samples is useful for various purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, the study of therapeutic targets in biological and pathological phenomena; and the comparative evaluation of new therapeutic target inhibitors.

[0307] In some embodiments, the present disclosure relates to a kit comprising a composition and specific instructions describing a method of using the composition in a subject requiring it to treat, improve, or prevent one or more pathological conditions and diseases (e.g., pruritus).

[0308] In this document, references to “about” of a value or parameter include (and describe) embodiments of said value or parameter itself. For example, a description referring to “about X” includes a description of “X”. In some embodiments, when used in connection with a measurement, or when used to modify a value, unit, constant, or range of values, the term “about” refers to a variation of + / - 10%, 5%, 2%, or 1%.

[0309] References to "between" two values ​​or parameters in the present invention include (and describe) embodiments that include said two values ​​or parameters themselves. For example, a description referring to "x to y" includes a description of "x" and "y" themselves.

[0310] Examples

[0311] The subject matter disclosed herein will be better understood by referring to the following examples, which are provided as examples of the invention, not as a limitation.

[0312] As indicated in the figure, the exemplary compound I (also referred to herein as compound 1) is as follows:

[0313]

[0314] As indicated in the figure, the exemplary compound II (also referred to herein as compound 2) is as follows:

[0315]

[0316] As indicated in the figure, the exemplary compound III (also referred to herein as compound 3) is as follows:

[0317]

[0318] Example 1

[0319] Detailed synthesis of exemplary compounds and intermediates as disclosed above is presented below in detail:

[0320] Table 1: List of compounds of formula (I) excluding isoandrographolide and andrographolide

[0321]

[0322]

[0323]

[0324]

[0325]

[0326]

[0327]

[0328]

[0329]

[0330]

[0331]

[0332] Representative synthesis of Compound 1 (also referred to herein as Compound I):

[0333]

[0334] Compound 1

[0335]

[0336] Step 1: Synthesis of Intermediate 1

[0337] 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-6-(hydroxymethyl)-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one in DCM (100 mL) IsoandrographolideTo a solution of (10.0 g, 28.5 mmol, 1.0 eq.), 2,2,6,6-tetramethylpiperidinyloxy(TEMPO, 0.892 g, 5.71 mmol, 0.2 eq.), tetrabutylammonium iodide (TBAI, 2.11 g, 5.71 mmol, 0.2 eq.), N-chlorosuccinimide (NCS, 5.72 g, 42.8 mmol, 1.5 eq.), aqueous NaHCO3 (0.5 M, 200 mL), and aqueous K2CO3 (0.05 M, 200 mL), cooled in an ice bath, were added. The resulting mixture was stirred at rt for 3 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the organic layer was separated, and the aqueous layer was extracted with DCM Tris (3 x 100 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was treated with hexane (100 mL) under stirring at rt for 15 min. The hexane layer was separated by gradient separation, and the obtained residue was treated with methanol (30 mL) at 0°C, stirred for 15 min, filtered, washed with cold methanol (10 mL), and dried under vacuum to obtain (2S,3aR,5aS,6S,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphthor[2,1-b]furan-6-carbaldehydride( Intermediate 1 ) was obtained as a grayish-white solid. Yield: 6.1 g (61.4%).

[0338] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 9.76 (d, J =2.0 Hz, 1H), 7.27 (q, J=2.0 Hz, 1H), 4.82-4.80 (m, 2H), 4.75-4.65 (m, 1H), 3.25-3.15 (m, 2H), 2.49-2.44 (m, 1H), 2.30-2.25 (m, 1H), 2.1-2.05 (m, 1H), 1.90-1.70 (m, 5H), 1.57-1.50 (m, 2H), 1.32 (s, 3H), 1.14-1.13 (m, 1H), 1.1 (s, 3H), 1.07-1.06 (m, 1H), 0.91 (s, 3H). 13C NMR (100 MHz, CDCl 3 , δ): 207.9, 172.5, 143.2, 138.3, 82.3, 73.2, 70.5, 59.2, 56.4, 53.6, 39.1, 36.3, 36.0, 32.9, 31.2, 27.8, 24.3, 19.8, 18.1, 13.7.

[0339] Step 1: Synthesis of Intermediate 1

[0340] DCM(5 L) 중 IsoandrographolideTo a solution of (500.0 g, 1.42 mol, 1.0 eq.), TEMPO (44.5 g, 0.28 mol, 0.2 eq.), TBAI (105.0 g, 0.28 mol, 0.2 eq.), NCS (285.0 g, 2.14 mol, 1.5 eq.), aqueous NaHCO3 (0.5 M, 10 L, 20 vol.), and aqueous K2CO3 (0.05 M, 10 L, 20 vol.), cooled under ice, were added. The resulting reaction mixture was stirred at rt for 3 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the organic layer was separated, and the aqueous layer was extracted twice (2 x 5 L) with dichloromethane. The combined organic layer was washed with a brine solution (5 L) and concentrated under reduced pressure. Acetonitrile (1 L) was added to the unpurified material and stirred at rt for 1 h. The precipitated solid was filtered, washed with acetonitrile (0.5 L) followed by n-heptane (1 L), and dried under vacuum to obtain (2S,3aR,5aS,6S,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carval( Intermediate 1 ) was provided as a grayish-white solid. Yield: 420.0 g (84.5%)

[0341] Step 2: Synthesis of Intermediate 2

[0342] Hydroxylamine hydrochloride (8.38 g, 121 mmol, 3.5 eq.) and pyridine (27.7 mL, 344 mmol, 10.0 eq.) in methanol (50 mL) were mixed with (2S,3aR,5aS,6S,7R, 9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbaldehyde ( Intermediate 1)(12.0 g, 34.4 mmol, 1.0 eq.) was added dropwise at rt to the solution. The resulting mixture was stirred at rt for 4 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated under reduced pressure, and ice-cold water (150 mL) was added under stirring. The precipitated product was filtered and dried under vacuum to obtain (E)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbaldehyde oxime( Intermediate 2 ) was provided as a grayish-white solid (9.0 g). The unrefined product was taken directly for the next step without further purification.

[0343] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.61 (brs, 1H), 7.44 (brs, 1H), 7.30-7.28 (m, 1H), 4.9-4.8 (m, 2H), 4.7-4.6 (m, 1H), 3.9-3.75 (m, 1H), 3.75-3.20 (m, 1H), 2.50 (t, J = 9.2 Hz, 1H), 2.26 (d, J =10.4 Hz, 1H), 2.15-2.0 (m, 1H), 1.98-1.75 (m, 4H), 1.73-1.5 (m, 7H), 1.33 (s, 3H), 1.14 (s, 3H), 1.10-1.05 (m, 2H), 0.97 (s, 3H). 13C NMR (100 MHz, CDCl 3 , δ): 172.5, 156.1, 143.1, 138.5, 82.5, 78.8, 73.3, 70.5, 56.9, 53.0, 44.4, 39.6, 36.0, 35.9, 32.8, 31.3, 27.6, 24.2, 18.1, 14.8.

[0344] Step 2: Synthesis of Intermediate 2

[0345] (2S,3aR,5aS,6S,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbaldehyde( Intermediate 1 )(420.0 g, 1.2 mol., 1.0 eq.) was added in fractions at rt to a solution of hydroxylamine hydrochloride (294.0 g, 4.21 mol., 3.5 eq.) and pyridine (0.97 L, 12.0 mol., 10.0 eq.) in methanol (5 L, 12 vol.). The resulting reaction mixture was stirred at rt for 4 h. The progress of the reaction was monitored by TLC and LC-MS. After the reaction was complete, ice-cold water (4.2 L) was added. The precipitated product was filtered and washed with heptane (1.26 L). The compound was exchanged with toluene under vacuum below 48°C (E)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbaldehyde oxime ( Intermediate 2 ) was provided as a grayish-white solid. Yield: 325.0 g. The unrefined product was taken directly for the next step without further purification.

[0346] Step 3: Synthesis of Compound 1

[0347] (E)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbaldehyde oxime in DCM (15 mL) Intermediate 2Triethylamine (1.96 mL, 14 mmol, 3.0 eq.) was added to a solution of )(1.7 g, 4.68 mmol, 1.0 eq.) at 0°C, followed by the addition of 4-toluenesulfonyl chloride (1.34 g, 7.02 mmol, 1.5 eq.) and a catalytic amount of 4-dimethylaminopyridine (57.1 mg, 0.468 mmol, 0.1 eq.). The resulting reaction mixture was heated to rt and stirred at rt for 3 hours. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with DCM (100 mL) and washed three times with water (3 x 30 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (100 to 200 mesh) using a mixture of ethyl acetate and hexane as the eluent (2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbonitrile( Compound 1 ) was provided as a white solid. Yield: 0.6 g (37.1%).

[0348] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.32 (q, J= 1.6Hz, 1H), 4.82 (q, J = 1.6 Hz, 2H), 4.8-4.7 (m, 1H), 3.3-3.2 (m, 1H), 2.50 (q, J = 8.0 Hz, 1H), 2.35-2.25 (m, 1H), 2.1-1.94 (m, 2H), 1.92-1.70 (m, 4H), 1.68-1.56 (m, 5H), 1.53 (s, 3H), 1.27 (s, 3H), 1.09-0.98 (m, 2H). 13C NMR (100 MHz, CDCl 3 , δ):172.9, 146.4, 136.2, 123.2, 82.4, 74.8, 72.5, 71.1, 56.8, 48.5, 42.8, 38.0, 35.8, 34.6, 32.3, 31.4, 28.2, 23.8, 19.4, 14.4. LC-MS: m / z 346.2 (M+H).

[0349] Step 3: Synthesis of Compound 1

[0350] (E)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbaldehyde oxime in dichloromethane (4.2 L, 12 vol.) Intermediate 2 PyBOP (1.0 kg, 1.92 mol., 2.0 eq.) was added to a solution of )(350.0 g, 0.96 mol., 1.0 eq.) at rt, followed by (at 5 min intervals) DBU (337.0 g, 2.21 mol., 2.3 eq.) (Observation: The reaction mixture became a clear, homogeneous solution). The resulting mixture was stirred at rt for 3 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was poured into water (10 vol.) and extracted twice (2 x 500 mL) with dichloromethane. The combined organic layer was washed with a saturated brine solution (3.5 L) and concentrated under reduced pressure. The obtained product was treated with ethyl acetate (3.5 L, 10 vol.) and stirred at rt for 2 h. The product was filtered, washed with ethyl acetate (700 mL), and dried under vacuum to obtain (2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbonitrile( Compound 1 ) was provided as a white solid. Yield : 215.0 g (64.7%).

[0351] Representative synthesis of Compound 2 (also referred to herein as Compound II):

[0352]

[0353] Compound 2

[0354]

[0355] Step 1: Synthesis of Intermediate 3

[0356] In pyridine (4.8 L, 2.4 vol.), 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-6-(hydroxymethyl)-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one( Isoandrographolide TBDMS-Cl (3.44 kg, 22.8 mol., 4.0 eq.) was added to a solution of )(2.0 kg, 5.7 mol., 1.0 eq.) at rt. The resulting mixture was stirred at rt for 3 h. The progress of the reaction was monitored by TLC and LCMS. After the reaction was complete, cold water (40 L) and heptane (20 L) were added to the reaction mixture and stirred at rt for 1 h, filtered, and washed with heptane (4 L). The compound was dried under vacuum to obtain 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-6-(((tert-butyl dimethylsilyl) oxy) methyl)-7-hydroxy-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one( Intermediate 3 )(1.1 kg) was provided as a grayish-white solid. The organic layer was combined, washed with cold water (20 L, 10 vol.) and a brine solution (20 L, 10 vol.), and concentrated under reduced pressure. The obtained crude product was treated with heptane (4 L) under stirring at 0°C for 1 h. The solid was filtered, washed with cold heptane (0.5 L), and dried under vacuum to obtain 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-6-(((tert-butyldimethylsilyl)oxy)methyl)-7-hydroxy-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one( Intermediate 3)(0.62 kg) was provided. Total yield: 1.72 kg(1.10 kg and 0.62 kg, 65.0%).

[0357] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.28 (t, J = 2.0 Hz, 1H), 4.82 (t, J = 2.4 Hz, 2H), 4.70 (m, 1H), 4.29-4.24 (m, 2H), 3.46 (d, J = 10.0 Hz, 1H), 3.35-3.25 (m, 1H), 2.46 (q, J =8.0 Hz, 1H), 2.21-2.17 (m, 1H), 2.05-2.01 (m, 1H), 1.82-1.68 (m, 4H), 1.57-1.40 (m, 4H), 1.25 (s, 3H), 1.13 (s, 3H), 0.97 (s, 3H), 0.91 (s, 9H), 0.1 (s, 6H).

[0358] Step 2: Synthesis of Intermediate 4

[0359] 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-6-(((tert-butyldimethylsilyl)oxy)methyl)-7-hydroxy-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one in DCM(20 L) Intermediate 3Pyridine (0.255 kg, 3.22 mol., 1.5 eq.), Celite (2 kg, 2 w / w), and pyridinium chlorochromate (0.927 kg, 4.3 mol, 2.0 eq.) were added at rt to a solution of )(1.0 kg, 2.15 mol., 1.0 eq.). The resulting mixture was stirred at rt for 24 h. The progress of the reaction was monitored by TLC and LCMS. After the reaction was complete, the reaction mixture was filtered over a Celite bed (2.0 kg), and the bed was washed with dichloromethane (5 L, 5 vol.). The filtrate (mL) was washed with water (10 L, 10 vol.) and a saturated saline solution (10 L, 10 vol.) and concentrated under reduced pressure. The obtained residue was dissolved in ethyl acetate (10 L, 10 vol.), charcoal (100.0 g, 10%) was added at rt, and heated at 50°C for 1 h. The resulting mixture was filtered and washed with ethyl acetate (5 L, 5 vol.). The filtrate was concentrated under reduced pressure. The unrefined product (brown solid) was treated with methanol (2 L, 2 vol.) under stirring at rt for 20 min, then cooled to 0°C and maintained for 2 h. The product was filtered, washed with cold methanol (0.5 L, 0.5 vol.), and dried under vacuum to obtain (2S,3aR,5aS,6R,9aR,9bS)-6-(((tert-butyldimethylsilyl)oxy)methyl)-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)decahydronaphtho[2,1-b]furan-7(3aH)-one( Intermediate 4) It was provided as a grayish-white solid. Yield: 650.0 g (65.2%).

[0360] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.31 (d, J =2.0 Hz, 1H), 4.83-4.77 (m, 3H), 3.84 (d, J = 10 Hz, 1H), 3.59 (d, J=10 Hz, 1H), 2.68-2.50 (m, 2H), 2.37-2.21 (m, 2H), 2.15-2.03 (m, 1H), 2.02-1.92 (m, 1H), 1.83-1.68 (m, 1H), 1.66-1.48 (m, 4H), 1.43-1.33 (m, 2H), 1.28 (s, 3H), 1.15 (s, 3H), 1.01 (s, 3H), 0.86 (s, 9H). 13C NMR (100 MHz, CDCl 3 , δ): 214.1, 172.5, 143.1, 138.4, 82.7, 73.1, 70.5, 66.5, 57.2, 53.9, 53.2, 39.1, 36.1, 35.9, 35.3, 33.0, 31.6, 25.8, 22.3, 19.3, 18.2, 15.5.

[0361] Step 3: Synthesis of Intermediate 5

[0362] (2S,3aR,5aS,6R,9aR,9bS)-6-(((tert-butyldimethylsilyl)oxy)methyl)-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)decahydronaphtho[2,1-b]furan-7(3aH)-one( Intermediate 4)(650.0 g, 1.4 mol., 1.0 eq.) was added dropwise at rt to a mixture of hydroxylamine hydrochloride (341.0 g, 4.9 mol., 3.5 eq.) and pyridine (1.14 L, 14.0 mol., 10.0 eq.) in methanol (7.8 L, 12 vol.). The resulting mixture was stirred at rt for 4 h. The progress of the reaction was monitored by TLC and LCMS. After the reaction was complete, ice-cold water (6.5 L) was added. The precipitated solid was filtered and washed with heptane (2.6 L, 4 vol.). The compound was distilled twice (2.6 L, 2 x 1 vol.) with toluene under reduced pressure. The crude product was treated with heptane (6.5 L, 10 vol.) while stirring at rt for 2 h. The product was filtered under vacuum, washed with heptane (1.3 L, 2 vol.), and dried to obtain 3-((2S,3aR,5aS,6S,9aR,9bS,E)-6-(((tert-butyldimethylsilyl)oxy)methyl)-7-(hydroxyimino)-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one( Intermediate 5 ) was provided as a grayish-white solid (598.0 g). The unrefined product was taken directly for the next step without further purification.

[0363] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.28 (d, J =2Hz, 1H), 4.81 (brs, 2H), 4.75 (t, J =9.6 Hz, 1H), 3.76 (d, J =6 Hz, 1H), 3.58 (d, J=10 Hz, 1H), 3.25-3.15 (m, 1H), 2.51-2.45 (m, 1H), 2.24-2.19 (m, 1H), 2.1-1.95 (m, 2H), 1.8-1.7 (m, 1H), 1.58-1.48 (m, 4H), 1.2 (s, 3H), 1.15 (s, 3H), 1.1 (s, 3H), 0.86 (s, 9H), 0.06 (s, 6H). 13C NMR (100 MHz, CDCl 3 , δ): 172.5, 163.4, 143.1, 138.4, 82.8, 73.1, 70.5, 66.9, 57.6, 53.6, 39.1, 36.3, 35.8, 32.9, 31.6, 25.8, 22.2, 19.0, 18.2, 17.8, 15.8.

[0364] Steps 4 and 5: Synthesis of Intermediate 6, Compound 2, and Intermediate 7

[0365] 3-((2S,3aR,5aS,6S,9aR,9bS,E)-6-(((tert-butyldimethylsilyl)oxy)methyl)-7-(hydroxyimino)-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one( Intermediate 5 4-toluenesulfonyl chloride (0.377 kg, 1.97 mol., 1.5 eq.) and potassium carbonate (0.546 kg, 3.95 mol., 3.0 eq.) were added to a solution of )(0.630 kg, 1.31 mol., 1.0 eq.) at 0°C. The reaction mixture was stirred at the same temperature for an additional 4 hours. The progress of the reaction was monitored by TLC and LCMS. After the reaction was complete, cold water (6.5 L, 10 vol.) was added to the reaction mixture and extracted with ethyl acetate (13 L, 2 x 10 vol.). The organic extracts were combined, washed with a brine solution (6.5 L, 10 vol.), and concentrated under reduced pressure. The obtained crude product was a light brown, thick syrup (0.6 kg). Intermediate 6 , Compound 2 , and Intermediate 7It was a mixture. The obtained crude product was dissolved in ethyl acetate (6 L, 10 vol.), and 4 M hydrochloric acid (3.14 L, 12.5 mol., 10 eq.) in 1,4-dioxane was added dropwise at 0°C. The resulting mixture was stirred at rt for 16 h. The progress of the reaction mixture was monitored by TLC. After the reaction was complete, ice-cold water (6 L, 10 vol.) was added, the organic layer was separated, and the aqueous layer was extracted with ethyl acetate (6 L, 10 vol.). The aqueous layer was stored separately. The combined organic layer was concentrated under reduced pressure. The obtained product was treated with ethyl acetate (1.2 L, 2 vol.) under stirring at rt for 1 h, the product was filtered, and washed with ethyl acetate (0.6 L, 1 vol.). The compound was dried under vacuum to obtain 3-((3aS,3bS,5aR,7S,8aS,8bR)-3a,5a,8b-trimethyl-3,3a,3b,4,5,5a,7,8,8a,8b,9,10-dodecahydrofuro[3',2':5,6]naphtho[2,1-c]isoxazole-7-yl)furan-2(5H)-one( Intermediate 7 ) was provided as a grayish-white solid; yield: 80.0 g (17.6%). The aqueous layer described above was adjusted to a pH of 8.5 to 9.0 using solid sodium carbonate (2.1 kg) and extracted with 10% methanol (6 L, 10 vol.) in dichloromethane. The organic layer was concentrated under reduced pressure. The obtained crude product was treated with ethyl acetate (3 L, 5 vol.) at 50°C under stirring for 2 h. The resulting mixture is cooled to rt, filtered, the obtained product is washed with ethyl acetate (1.2 L, 2 vol.) followed by heptane (1.2 L, 2 vol.), and dried under vacuum to obtain (2S,3aR,5aS,6R,10aR,10bS)-6-(hydroxymethyl)-3a,6,10a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydro-8H-benzofuro[5,4-c]azepine-8-one( Compound 2) was provided as a grayish-white solid. Yield: 142.0 g (31%).

[0366] Intermediate 7 : 1 ¹H NMR (400 MHz, DMSO-d 6 , δ): 7.46 (q, J =2.0 Hz, 1H), 4.85 (t, J =2.4 Hz, 2H), 4.54-4.50 (m, 1H), 3.94 (d, J =8.4 Hz, 1H), 3.83 (d, J =8.4 Hz, 1H), 2.62-2.53 (m, 1H), 2.46-2.40 (m, 1H), 2.34-2.27 (m, 1H), 2.13-2.01 (m, 2H), 1.85-1.76 (m, 1H), 1.61 (d, J =7.2 Hz, 1H), 1.56-1.45 (m, 2H), 1.43-1.32 (m, 2H), 1.26-1.18 (m, 1H), 1.13 (s, 3H), 1.05 (s, 3H), 1.05 (s, 3H). 13C NMR (100 MHz, DMSO-d 6 , δ): 172.4, 164.9, 136.3, 81.9, 76.9, 71.9, 70.6, 54.3, 52.0, 49.9, 38.8, 35.9, 35.3, 34.0, 32.2, 30.8, 25.3, 18.8, 16.9, 13.5. LC-MS: m / z 346.2 (M+H).

[0367] Compound 2 : 1 ¹H NMR (400 MHz, DMSO-d 6 , δ): 7.48 (d, J =2.0 Hz, 1H), 6.47 (brs, 1H), 4.86 (brs, 2H), 4.82 (t, J = 5.2 Hz, 1H), 4.50 (t, J =7.6 Hz, 1H), 3.59-3.45 (m, 2H), 2.34-2.29 (m, 2H), 2.20 (q, J=7.6 Hz, 1H), 2.02-1.92 (m, 2H), 1.79-1.74 (m, 1H), 1.72-1.59 (m, 2H), 1.58-1.46 (m, 3H), 1.42-1.33 (m, 1H), 1.18 (s, 3H), 1.13 (s, 3H), 1.04 (s, 3H). 13C NMR (100 MHz, DMSO-d 6 , δ): 172.2, 172.4, 146.1, 135.4, 82.2, 71.6, 70.7, 65.6, 60.2, 54.7, 50.3, 38.5, 35.7, 34.8, 32.7, 30.9, 30.9, 28.2, 20.4, 17.53. LC-MS: m / z 364.2 (M+H)

[0368] Representative synthesis of Compound 3 (also referred to herein as Compound III):

[0369]

[0370] Compound 3

[0371]

[0372] Step 1: Synthesis of Intermediate 8:

[0373] (2S,3aR,5aS,6R,10aR,10bS)-6-(hydroxymethyl)-3a,6,10a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydro-8H-benzofuro[5,4-c]azepine-8-one in (600 mL) of DCM Compound 2DMP (87.5 g, 206 mmol, 2.5 eq.) was added at rt to a solution of )(30.0 g, 82.5 mmol, 1.0 eq.). The resulting mixture was stirred at rt for 3 h. The progress of the reaction mixture was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with DCM (100 mL) and washed twice with saturated hypo solution (2 x 100 mL), saturated NaHCO3 solution (100 mL), and water (100 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was ground in methanol (90 mL) under stirring at rt for 30 min. The product is filtered, washed with cold methanol (10 mL), and dried under reduced pressure to obtain (2S,3aR,5aS,6R,10aR,10bS)-3a,6,10a-trimethyl-8-oxo-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydro-2H-benzofuro[5,4-c]azepine-6-carbaldehyde( Intermediate 8 ) was provided as a white solid. Yield: 29.0 g (97.2%).

[0374] Intermediate 8 : 1 1H NMR (400 MHz, CDCl₃ 3 , δ) : 9.76 (s, 1H), 7.31 (q, J =2.0 Hz, 1H), 5.81 (brs, 1H), 4.83 (t, J =2.4 Hz, 2H), 4.72-4.68 (m, 1H), 2.45 (q, J =8.0 Hz, 1H), 2.40-2.35 (m, 2H), 2.32-2.25 (m, 1H), 2.15-1.85 (m, 4H), 1.78-1.72 (m, 1H), 1.7-1.6 (m, 3H), 1.43 (s, 3H), 1.2 (s, 3H), 1.05 (s, 3H). 13C (NMR 100 MHz, CDCl 3 , δ):199.0, 178.6, 172.4, 143.4, 137.7, 82.2, 72.9, 70.5, 66.2, 56.4, 55.0, 39.6, 36.4, 35.5, 33.4, 31.2, 30.5, 25.3, 20.4, 16.2.

[0375] Step 2: Synthesis of Intermediate 9

[0376] (2S,3aR,5aS,6R,10aR,10bS)-3a,6,10a-trimethyl-8-oxo-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydro-2H-benzofuro[5,4-c]azepine-6-carbaldehyde in methanol (350 mL) Intermediate 8 ) A pre-dissolved solution of hydroxylamine hydrochloride (8.07 g, 116 mmol, 1.5 eq.) and pyridine (62.4 mL, 775 mmol, 10.0 eq.) in methanol (200 mL) was added dropwise at RT to a solution of (28.0 g, 77.5 mmol, 1.0 eq.). The resulting mixture was stirred at RT for 3 h. The progress of the reaction mixture was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated under reduced pressure, and ice-cold water (500 mL) was added. The precipitated product was filtered and dried under vacuum to obtain (E)-3a,6,10a-trimethyl-8-oxo-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydro-2H-benzofuro[5,4-c]azepine-6-carbaldehyde oxime ( Intermediate 9 ) It was provided as a grayish-white solid (19.0 g). The unrefined product was taken directly for the next step without further purification.

[0377] 1 1H NMR (400 MHz, CDCl₃ 3 , δ) : 8.79 (d, J =2.8 Hz, 1H), 7.53 (s, 1H), 7.30 (q, J =2.0 Hz, 1H), 5.59 (brs, 1H), 4.82 (t, J=2.4 Hz, 2H), 4.68 (t, J =2.8Hz, 1H), 2.65-2.53 (m, 1H), 2.46 (q, 7.6 Hz, 1H), 2.35 (q, J =7.6 Hz, 1H), 2.25-2.17 (m, 1H), 2.10-1.90 (m, 1H), 1.7-1.6 (m, 5H), 1.55-1.5 (m, 1H), 1.48-1.38 (m, 4H), 1.19 (s, 3H), 0.9 (s, 3H). 13C NMR (100 MHz, CDCl 3 , δ): 179.7, 172.3, 151.6, 143.4, 137.7, 82.3, 72.8, 70.5, 58.7, 56.5, 54.2, 39.7, 36.6, 35.0, 33.2, 31.2, 30.8, 30.7, 20.3, 15.6.

[0378] Step 3: Synthesis of Compound 3

[0379] (E)-3a,6,10a-trimethyl-8-oxo-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydro-2H-benzofuro[5,4-c]azepine-6-carbaldehyde oxime in DCM (600 mL) Intermediate 9Et3N (21.1 mL, 151 mmol, 3.0 eq.) was added to a solution of )(19.0 g, 50.5 mmol, 1.0 eq.), the reaction mixture was cooled to 0°C, and then 4-methylbenzene-1-sulfonyl chloride (14.4 g, 75.7 mmol, 1.5 eq.) and DMAP (0.617 g, 5.05 mmol, 0.1 eq.) were added. The resulting mixture was brought to rt and stirred at rt for 3 h. The progress of the reaction mixture was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with DCM (300 mL) and washed with water (3 x 100 mL). The separated organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was treated with methanol (100 mL) and stirred at 0°C for 30 min. The product was filtered, washed with cold methanol (10 mL), and dried under vacuum to obtain (2S,3aR,5aS,6R,10aR,10bS)-3a,6,10a-trimethyl-8-oxo-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydro-2H-benzofuro[5,4-c]azepine-6-carbonitrile( Compound 3 ) was provided as a grayish-white solid. Yield: 17.0 g (93.97%).

[0380] 1 1H NMR (400 MHz, CDCl₃ 3, δ) : 7.33 (q, 2.0 Hz, 1H), 5.95 (brs, 1H), 4.83-4.82 (m, 2H), 4.70-4.66 (m, 1H), 2.98-2.91 (m, 1H), 2.47-2.41 (m, 2H), 2.20-2.15 (m, 1H), 2.05-1.85 (m, 2H), 1.80-1.63 (m, 7H), 1.42-1.38 (m, 5H), 1.24 (s, 3H). 13C NMR (100 MHz, CDCl 3 , δ):177.7, 172.3, 144.0, 136.5, 120.5, 82.1, 72.6, 70.5, 58.0, 52.1, 51.5, 40.0, 36.9, 33.2, 32.8, 31.1, 30.3, 29.6, 20.9, 14.0. LC-MS: m / z 359.2 (M+H).

[0381] Representative synthesis of reductive amination products (compounds 4 to 14):

[0382]

[0383] Representative synthesis of Compound 4:

[0384]

[0385] Compound 4

[0386] (2S,3aR,5aS,6S,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbaldehyde in THF (10 mL) Intermediate 1 Benzylamine (533 μL, 4.8 mmol, 1.7 eq.) and acetic acid (catalyst amount, 82 μL) were added at rt to a stirred solution of )(1 g, 2.87 mmol, 1 eq.). After 30 min, NaCNBH3 (361 mg, 5.74 mmol, 2 eq.) was added, and the resulting reaction mixture was stirred at rt for 12 h. The progress of the reaction was monitored by TLC, the reaction mixture was diluted with water (50 mL), and extracted with EtOAc (3 x 40 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in DCM (100 to 200 silica gel mesh) using MeOH as the eluent to obtain 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-6-((benzylamino)methyl)-7-hydroxy-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one( Compound 4) was provided as a grayish-white solid. Yield: 430 mg (34%).

[0387] 1 ¹H NMR (400 MHz, CDCl3, δ): 7.36-7.27(m, 6H), 4.80 (d, J = 4.4 Hz, 2H), 4.67(t, J = 8.0 Hz, 1H), 3.76(q, J = 12.8 Hz, 2H), 3.31( dd, J =11.6, 3.2 , 1H), 3.11(d, J = 12.0 Hz, 1H), 2.72(dd, J = 12.0, 0.8, Hz, 1H), 2.41(q, J = 8.0 Hz, 1H), 2.15(d, J = 12.4, 5.2 Hz, 1H), 2.05- 1.96(m, 1H), 1.75-1.67(m, 2H), 1.53- 1.45(m, 5H), 1.24(bs, 3H), 1.10(bs, 3H), 1.04 (d, J = 3.6 Hz, 1H), 0.99 (d, J = 3.6 Hz, 1H), 0.96 (d, J = 3.2 Hz, 1H), 0.90 (s, 3H). 13C NMR (100 MHz, CDCl3, δ): 175.1, 172.6, 161.2, 158.5, 143.2, 138.3, 137.3, 128.6, 127.5, 82.72, 80.4, 73.1, 70.8, 58.33, 54.3, 53.4, 50.7, 46.3, 40.5, 39.2, 35.8, 32.8, 31.5, 27.8, 23.1, 18.4, 16.3.

[0388] Representative synthesis of Compound 5:

[0389]

[0390] Compound 5

[0391] (2S,3aR,5aS,6S,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbaldehyde in THF (10 mL) Intermediate 1 2-chlorobenzylamine (417 μL, 3.44 mmol, 1.2 eq.) and acetic acid (catalyst amount) (49 mg) were added at rt to a stirred solution of ) (1 g, 2.87 mmol, 1 eq.). After 30 min, NaCNBH3 (361 mg, 5.74 mmol, 2 eq.) was added at rt, and the resulting reaction mixture was stirred at rt for 12 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with water (30 mL) and extracted with EtOAc (3 x 40 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in DCM (100 to 200 silica gel mesh) using MeOH as the eluent to obtain 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-6-(((2-chlorobenzyl)amino)methyl)-7-hydroxy-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one( Compound 5 ) was provided as a grayish-white solid. Yield: 50 mg (3%).

[0392] 1 ¹H NMR (400 MHz, CDCl3, δ): 7.38 ( d, J =4 Hz, 2H ), 7.28 - 7.25 (m, 3H), 4.81 (s, 2H), 4.69 (t, J = 7.2 Hz, 1H), 3.91 (q, J =13.6, 2H), 3.33 (dd, J =11.6, 3.6Hz, 2H), 3.12 (d, J =12 Hz, 1H), 2.72 (d, J=10.8Hz, 1H), 2.43(q, J = 8.0, 1H), 2.17 (dt, J =0 3.6 Hz, Hz, 1H), 2.05-2.01 (m, 1H), 1.78-1.69 (m, 2H), 1.54- 1.46 (m, 5H), 1.26 (s, 3H), 1.11 (s, 3H), 1.00 (d, J = 4.4 Hz, 1H) 0.94 (s, 3H), 0.90 (dd, J =10.2, 3.2Hz, 2H). 13C NMR (100 MHz, CDCl3, δ): 172.5, 143.1, 138.2, 135.0, 133.9, 130.8, 129.6, 129.1, 127.02, 82.6, 73.0, 70.4, 58.2, 53.1, 51.4, 50.4, 50.4, 39.0, 35.9, 35.7, 32.7, 31.45, 27.6, 23.0, 18.3, 16.2.

[0393] Representative synthesis of Compound 6:

[0394]

[0395] Compound 6

[0396] To a solution of intermediate 1 (1 g, 2.87 mmol, 1 eq.) in THF (10 mL), 3-fluorobenzylamine (0.4 mL, 2.87 mmol, 1.2 eq.) and acetic acid (172 mg) were added at rt. After 4 h, methanol (5 mL) and NaCNBH3 (361 mg, 5.74 mmol, 2 eq.) were added. The resulting mixture was stirred at rt for 12 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with water (15 mL) and extracted with EtOAc (2 x 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in DCM (60 to 120 silica gel mesh) using MeOH as the eluent to obtain the desired product. Compound 6 It was provided as a grayish-white solid. Yield: 210 mg (15%).

[0397] 1 ¹H NMR (400 MHz, CDCl3, δ ): 7.32-7.27 (m, 1H), 7.27(s, 1H), 7.06 (d, J =8Hz, 1H), 7.00-6.94 (m, 2H), 4.80 (t, J = 2.8Hz, 2H), 4.68 (t, J =7.6 Hz, 1H), 3.73 (d, J = 2.8 Hz, 2H), 3.30(dd, J =10.4, 4.0 Hz, 1H), 3.09(d, J = 12.0 Hz, 1H), 2.70(d, J = 10.4 Hz, 1H), 2.42(q, J =8.0 Hz, 1H), 2.16(dt, J = 14.4, 3.2 Hz, 1H), 2.04-1.96 (m, 1H), 1.79-1.68(m, 2H), 1.61-1.45(m, 6H), 1.24(s, 3H), 1.21(d, J = 5.2 Hz, 1H), 1.16(d, J = 3.2 Hz, 1H), 1.10 (s, 3H), 1.00(td, J = 12.8, 2.4 Hz, 1H), 0.93(s, 3H), 0.89(t, J = 4.8 Hz, 1H), 0.86(d, J = 4.0 Hz, 1H), 13C NMR (100 MHz, CDCl3, δ): 177.2, 172.4, 164.0, 161.5, 143.0, 141.3, 138.3, 130.0, 123.7, 115.1, 114.4, 82.6, 80.4, 73.0, 70.4, 58.2, 54.0, 53.2, 50.9, 46.6, 40.6, 39.6, 36.0, 35.8, 32.9, 31.4, 27.8, 23.0, 18.3, 16.4.

[0398] Representative synthesis of Compound 7:

[0399]

[0400] Compound 7

[0401] 2-fluorobenzylamine (431 mg, 3.44 mmol, 1.2 eq.) and acetic acid (172 mg) were added at rt to a solution of intermediate 1 (1 g, 2.87 mmol, 1 eq.) in THF (10 mL). After 4 h, methanol (5 mL) and NaBH4 (271 mg, 5.74 mmol, 2 eq.) were added. The resulting mixture was stirred at rt for an additional 12 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with water (15 mL) and extracted with DCM (3 x 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography using MeOH as the eluent in DCM (60 to 120 silica gel mesh) to provide compound 7 as a grayish-white solid, yield: 300 mg (21%).

[0402] 1 ¹H NMR (400 MHz, CDCl3, δ): 7.30-7.27 (m, 2H), 7.244 (d, J =3.2Hz, 1H), 7.12 (dd, J =12.0Hz, 7.6Hz, 1H), 7.09 (m, 1H), 4.80 (t, J =2.8Hz, 2H), 4.68 (t, J =2.8Hz, 1H), 3.76 (s, 2H), 3.29 (dt, J =8.4Hz, 3.2Hz, 1H), 3.08 (d, J =12Hz, 1H), 2.69 (dd, J =12Hz, 1H), 2.43 (q, J =8.0Hz, 1H), 2.15 (dt, J=14.4Hz, 4.4Hz, 1H), 2.03-1.97 (m, 1H), 1.77-1.70 (m, 3H), 1.53 (t, J =5.6Hz, 2H), 1.48-1.41 (m, 3H), 1.25 (d, J =3.2Hz, 1H), 1.22 (s, 3H), 1.09 (s, 3H), 1.00 (d, J =0Hz, 1H), 0.95 (s, 3H), 0.89 (q, J =4.8Hz, 1H) . 13C NMR (100 MHz, CDCl3, δ): 172.5, 162.3, 159.9, 143.0, 138.4, 130.5, 129.2, 125.8, 124.2, 115.4, 82.6, 80.4, 73.1, 70.4, 58.3, 53.3, 50.8, 47.8, 40.6, 37.2, 36.1, 32.8, 31.5, 29.6, 27.9, 23.0, 18.40, 16.2.

[0403] Representative synthesis of Compound 8:

[0404]

[0405] Compound 8

[0406] 2-methylbenzylamine (417 mg, 3.44 mmol, 1.2 eq.) and acetic acid (172 mg) were added at rt to a solution of intermediate 1 (1 g, 2.87 mmol, 1 eq.) in THF (10 mL). After 4 h, methanol (5 mL) and NaBH4 (217 mg, 5.74 mmol, 2 eq.) were added. The resulting mixture was stirred at rt for an additional 12 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with water (10 mL) and then extracted with DCM (2 x 10 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in DCM (60 to 120 silica gel mesh) using MeOH as the eluent. Compound 8It was provided as a grayish-white solid. Yield: 300 mg (21%).

[0407] 1 ¹H NMR (400 MHz, CDCl3, δ): 7.27 (t, J =2.0Hz, 1H), 7.21-7.16 (m, 4H), 4.80 (t, J =7.6, 1H), 3.72 (s, 2H), 3.28 (dd, J =11.6Hz, 2.4Hz, 1H), 3.14 (d, J =12.0Hz, 1H), 2.75 (d, J =11.6Hz, 1H), 2.43 (q, J =7.6Hz, 1H), 2.34 (s, 3H), 2.18 (dd, J =10.8Hz, 3.6 Hz, 1H), 2.04-1.96 (m, 1H), 1.77-1.69 (m, 3H), 1.55-1.45 (m, 6H), 1.25 (t, J =2.0, 1H), 1.23 (s, 3H), 1.10 (s, 3H), 1.01 (dd, J =13.2Hz, 3.2Hz, 1H), 0.96 (s, 3H). 13C NMR (100 MHz, CDCl3, δ): 172.5, 143.0, 138.3, 136.5, 136.4, 130.4, 128.9, 127.5, 125.9, 82.6, 80.3, 73.1, 70.4, 58.2, 53.2, 52.2, 51.2, 40.6, 39.1, 36.1, 35.8, 32.8, 31.5, 27.8, 23.0, 18.9, 18.4, 16.3.

[0408] Representative synthesis of Compound 9:

[0409]

[0410] Compound 9

[0411] In THF (10 mL) Intermediate 13-chlorobenzylamine (610 mg, 4.34 mmol, 1.5 eq.) and acetic acid (172 mg) were added at rt to a solution of (1 g, 2.87 mmol, 1 eq.). After 30 min, NaBH3CN (361 mg, 5.74 mmol, 2 eq.) was added. The resulting mixture was stirred at rt for an additional 12 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with water (10 mL) and then extracted with DCM (2 x 10 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in DCM (60 to 120 silica gel mesh) using MeOH as the eluent. Compound 9 It was provided as a grayish-white solid. Yield: 340 mg (24%).

[0412] 1 ¹H NMR (400 MHz, CDCl3, δ): 7.28 (m, 2H), 7.24 (d, J =2.0Hz, 1H), 7.20-7.16 (m, 1H), 4.80 (t, J =2.8Hz, 2H), 4.70 (t, J =7.6Hz, 1H), 3.71 (s, 2H), 3.30 (dd, J =10.4Hz, 2.8Hz, 1H), 3.08 (d, J =12.0Hz, 1H), 2.70 (dd, J =12.0Hz, 1.2Hz, 1H), 2.43 (q, J =8.0Hz, 1H), 2.19 (dd, J=6.4Hz, 3.2Hz, 1H), 2.04- 1.96 (m,1H), 1.79-1.69 (m, 3H), 1.61- 1.41 (m, 6H), 1.29 (d, J =2.8Hz, 1H), 1.24 (s, 3H), 1.10 (s, 3H), 1.02(dd, J =2.8Hz, 1H), 0.97 (s, 3H), 0.93 (d, J=6.4Hz, 1H). 13C NMR (100 MHz, CDCl3, δ): 172.5, 143.0, 140.8, 138.3, 134.3, 129.8, 128.3, 127.5, 126.3, 82.6, 80.4, 73.1, 58.2, 54.2, 53.2, 51.0, 40.6, 39.2, 36.1, 35.8, 32.8, 31.5, 27.8, 23.08, 18.4, 16.3.

[0413] Representative synthesis of Compound 10:

[0414]

[0415] Compound 10

[0416] In THF (10 mL) Intermediate 1 4-fluorobenzylamine (431 mg, 3.44 mmol, 1.2 eq.) and acetic acid (0.164 mL, 2.87 mmol, 1 eq.) were added at rt to a solution of (1 g, 2.87 mmol, 1 eq.). After 30 min, NaBH3CN (361 mg, 5.74 mmol, 2 eq.) was added. The resulting mixture was stirred at rt for an additional 12 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with water (10 mL) and then extracted with DCM (2 x 10 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in DCM (60 to 120 silica gel mesh) using MeOH as the eluent. Compound 10 It was provided as a grayish-white solid. Yield: 310 mg (23%).

[0417] 1 ¹H NMR (400 MHz, CDCl3, δ): 7.26-7.22(m, 3H), 7.00 (t, J = 8.8 Hz, 2H), 4.79(t, J = 2Hz, 2H), 4.67(t, J = 8.0 Hz, 1H), 3.68(q, J= 12.8 Hz, 2H), 3.28( dd, J =11.2, 2.4 , 1H), 3.07(d, J = 12.0 Hz, 1H), 2.68 (d, J = 12.0 Hz, 1H), 2.41(q, J = 8.0 Hz, 1H), 2.16(dd, J = 10.4, 3.2 Hz, 1H), 2.00- 1.97( m, 1H), 1.75-1.87( m, 2H), 1.52- 1.45( m, 5H), 1.22(s, 3H), 1.09(s, 3H), 1.04 (d, J = 3.6 Hz, 1H), 0.99 (d, J = 3.6 Hz, 1H), 0.96 (d, J = 3.2 Hz, 1H), 0.91 (s, 3H), 0.87(d, J = 3.6 Hz, 1H). 13C NMR (100 MHz, CDCl3, δ): 172.5, 163.3, 160.9, 143.1, 138.4, 134.6, 134.5, 129.9, 115.4, 115.2, 82.7, 80.5, 73.1, 70.5, 58.3, 53.3, 50.8, 40.66 39.25 36.15 35.87 32.87 31.57 29.69 27.95 23.12 18.47 16.35.

[0418] Representative synthesis of Compound 11:

[0419]

[0420] Compound 11

[0421] THF(8 mL) 중 Intermediate 1Cyclohexylamine (0.123 mL, 1.08 mmol, 1.5 eq.) and acetic acid (catalyst amount) (0.02 mL) were added at rt to a solution of (250 mg, 0.717 mmol, 1 eq.). After 1 h, NaCNBH3 (90.2 mg, 1.43 mmol, 2 eq.) was added. The resulting mixture was stirred at rt for 5 h. The progress of the reaction was monitored by TLC. After the reaction was complete, water (25 mL) was added to the reaction mixture and extracted with EtOAc (3 x 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in DCM (100 to 200 silica gel mesh) using methanol as the eluent. Compound 11 It was provided as a white solid. Yield: 70 mg (22%).

[0422] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.29 (t, J=4.0 Hz, 1H), 4.83 (d, 2H), 4.71 (d, J=15.2 Hz, 1H), 3.34-3.31 (dd, 1H), 3.14 (d, 1H), 2.72 (d, J=11.6 Hz, 1H), 2.72 (d, 1H), 2.48-2.38 (m, 2H), 2.21-2.17 (t, 1H), 2.03-2.00 (m, 2H), 1.95-1.48 (m, 10H), 1.27-1.25 (d, 4H), 1.12 (s, 3H), 1.01 (s, 3H), 0.96-0.89 (m, 1H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ): 172.5, 143.1, 138.4, 82.7, 80.5, 73.1, 70.5, 58.4, 57.1, 53.3, 48.2, 40.3, 39.3, 36.1, 35.8, 32.2, 33.0, 32.8, 31.6, 28.1, 25.9, 24.8, 24.8, 23.2, 18.4, 16.5.

[0423] Representative synthesis of Compound 12:

[0424]

[0425] Compound 12

[0426] In THF (8 mL) Intermediate 1 Cyclopropylamine (0.0597 mL, 0.861 mmol, 1.5 eq.) was added to a solution of (200 mg, 0.574 mmol, 1 eq.), and acetic acid (catalyst amount) (0.01 mL) was added at rt. After 1 h, NaCNBH3 (72.1 mg, 1.15 mmol, 2 eq.) was added. The resulting mixture was stirred at rt for 5 h. The progress of the reaction was monitored by TLC. After the reaction was complete, water (25 mL) was added to the reaction mixture and extracted with EtOAc (3 x 30 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in DCM (100 to 200 silica gel mesh) using methanol as the eluent. Compound 12 was provided as a white solid. Yield: 70 mg (31%).

[0427] 1 ¹H NMR (400 MHz, DMSO, δ): 7.46 (d, J =1.2 Hz, 1H), 4.86 (s, 2H), 4.51 (t, J =15.2 Hz, 1H), 3.35-3.23 (m, 3H), 2.88 (d, 1H), 2.60-2.50 (brs, 1H), 2.25-2.20 (m, 1H), 2.08-1.97 (m, 2H), 1.67-1.59 (m, 3H), 1.55-1.41 (m, 3H), 1.14 (s, 3H), 1.06 (s, 3H), 1.03-0.99 (m, 2H), 0.88 (s, 4H), 0.72-0.66 (m, 3H). 13 13C NMR (100 MHz, DMSO, δ):172.9, 146.2, 136.5, 82.4, 78.4, 72.5, 71.2, 57.6, 52.2, 50.8, 38.4, 35.9, 35.5, 32.6, 31.7, 31.4, 27.1, 23.1, 18.3, 16.1.

[0428] Representative synthesis of Compound 13:

[0429]

[0430] Compound 13

[0431] In THF (8 mL) Intermediate 1 Butane-1-amine (0.111 mL, 1.12 mmol, 1.3 eq.) and acetic acid (catalyst amount) (0.03 mL) were added at rt to a solution of (300 mg, 0.861 mmol, 1 eq.). After 1 h, NaCNBH3 (108 mg, 1.72 mmol, 2 eq.) was added. The resulting mixture was stirred at rt for 5 h. The progress of the reaction was monitored by TLC. After the reaction was complete, water (25 mL) was added to the reaction mixture and extracted with EtOAc (3 x 25 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by recrystallization using MeOH:DCM (9:1) (5 mL). The solid was filtered and dried under vacuum Compound 13 It was provided as a white solid. Yield: 110 mg (31%).

[0432] 1 ¹H NMR (400 MHz, DMSO, δ): 7.47 (s, 1H). 4.68 (d, 2H0, 3.19 (q, 1H), 2.93 (q, 2H0, 2.81 (d, 1H), 2.25-2.20 (q, 1H), 2.08-1.98 (m, 2H), 1.64-1.47 (m, 8H), 1.39 (br, s, 2H), 1.35-1.28 (m, 2H), 1.17 (s, 3H), 1.06-1.07 (s, 4H), 0.9-0.8 (q, 6H). 13C NMR (100 MHz, DMSO, δ):172.9, 146.2, 136.5, 82.4, 78.2, 72.5. 71.1, 57.5, 52.0, 49.2, 48.1, 38.2, 35.8, 35.4, 32.5, 31.4, 27.5, 26.8. 22.8, 19.7, 18.2, 16.1, 13.9.

[0433] Representative synthesis of Compound 14:

[0434]

[0435] Compound 14

[0436] In THF (3 mL) Intermediate 1 1-(4-chlorophenyl)methaneamine (48.8 mg, 0.344 mmol, 1.2 eq.) and acetic acid (catalyst amount) (0.01 mL) were added at rt to a solution of (100 mg, 0.287 mmol, 1 eq.). After 30 min, NaCNBH3 (36.1 mg, 0.574 mmol, 2 eq.) was added. The resulting mixture was stirred at rt for an additional 12 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with water (15 mL) and extracted with EtOAc (3 x 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in DCM (100 to 200 silica gel mesh) using MeOH as the eluent. Compound 14 It was provided as a grayish-white solid. Yield: 70 mg (51%).

[0437] 1 1H NMR (400 MHz, CDCl₃ 3 , δ):7.35-7.28 (m, 4H), 4.05 (d, 1 H), 3.85 (d, 1H), 3.44 (dd, 1H), 3.19 (d, 1 H), 2.88 (d, 1H), 2.41 (q, 1 H), 2.19 (t, 1H), 2.06-1.98 (m, 2H), 1.71 (d, 2H), 1.56-1.38 (m, 6H), 1.36-1.27 (s, 6H), 1.25-1.18 (m, 4H), 1.16-1.02 (m, 2H), 0.99-0.81 (m, 4H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ): 172.5, 143.2, 138.2, 134.9, 131.6, 130.8, 129.2, 82.5, 79.5, 73.1, 70.5, 58.1, 52.8, 52.0, 49.5, 40.3, 38.7, 35.9, 35.6, 32.8, 31.4, 29.6, 26.7, 22.6, 18.2, 16.1.

[0438] Representative synthesis of Compound 15:

[0439]

[0440] Compound 15

[0441]

[0442] THF(8 mL) 중 Intermediate 8Cyclopropylamine (0.072 mL, 1.04 mmol, 1.5 eq.) and acetic acid (catalyst amount) (0.1 mL) were added at rt to a solution of (250 mg, 0.692 mmol, 1 eq.). After 1 h, NaCNBH3 (86.9 mg, 1.38 mmol, 2 eq.) was added. The resulting mixture was stirred at rt for 12 h. The progress of the reaction was monitored by TLC. After the reaction was complete, water (25 mL) was added to the reaction mixture and extracted with EtOAc (3 x 25 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by flash column chromatography using MeOH in DCM as the eluent, Compound 15 It was provided as a white solid. Yield: 80 mg (28%).

[0443] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.36-7.28 (q, 1H), 5.92 (s, 1H), 4.82 (s, 2H), 4.69 (t, 3H), 3.08 (d, 1H), 2.83 (d, 1H), 2.49-2.42 (m, 4H), 2.16-1.99 (m, 2H), 1.79-1.51 (m, 9H), 1.27-1.19 (m, 10H), 0.42 (d, 2H), 0.31 (t, 2H). 13C NMR (100 MHz, CDCl 3 , δ): 176.8, 172.5, 143.6, 137.6, 82.6, 72.7, 70.5, 60.0, 56.3, 5b4.9, 51.5, 39.3, 36.8, 35.1, 33.5, 31.3, 31.2, 30.8, 29.5, 20.9, 18.4, 6.8, 6.6.

[0444] Representative synthesis of reductive amination products (compounds 16 and 17):

[0445]

[0446] Representative synthesis of Compound 16:

[0447]

[0448] Compound 16

[0449] Step 1: Synthesis of Intermediate 10

[0450] In pyridine (10 mL) at rt Intermediate 1 Acetic anhydride (6 mL) was added dropwise at rt to a solution of (6.0 g, 17.2 mmol, 1 eq.). The resulting mixture was stirred at rt for 3 h. The progress of the reaction was monitored by TLC. After the reaction was complete, ice-cold water (100 mL) was added, the precipitated solid was filtered, washed with water (100 mL), and dried under vacuum. Intermediate 10 It was provided as a grayish-white solid. Yield: 5.6 g (83.29%).

[0451] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 10.10 (s,1H), 7.28 (t, J=3.2 Hz, 1H), 4.82 (s, 2H), 4.73-4.69 (q, 2H), 2.50-2.44 (q, 1H), 2.24-2.19 (t, 1H), 2.07-2.04 (t, 4H), 2.01-1.96 (m, 1H), 1.90-1.86 (dd, 1H), 1.75-1.70 (t, 1H), 1.63-1.61 (d, 1H), 1.57-1.39 (m, 2H), 1.24-1.21 (d, 2H), 1.30 (s, 3H), 0.93 (s, 3H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ): 203.9, 172.5, 170.4, 143.2, 138.2, 82.4, 78.5, 73.1, 70.5, 56.6, 54.4, 51.5, 38.3, 35.7, 35.6, 33.1, 31.3, 23.8, 2123, 21.0, 18.3, 16.9.

[0452] Step 2: Synthesis of Intermediate 11

[0453] In tert-butanol (180 mL), 2-methyl-2-butene (20.9 mL, 197 mmol, 14 eq.) and water (140 mL) Intermediate 10 NaOCl2 (5.1 g, 56.3 mmol, 4 eq.) and NaH2PO4 (6.65 g, 56.3 mmol, 4 eq.) were added at rt to a solution of (5.5 g, 14.1 mmol, 1 eq.). The resulting mixture was stirred at rt for 48 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with EtOAc (200 mL). The organic layer was separated and washed with water (3 x 80 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in hexane using EtOAc (100 to 200 silica gel mesh) as the eluent. Intermediate 11 It was provided as a white solid. Yield: 3.7 g (64.62%).

[0454] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.29 (t, J=6.8 Hz, 1H), 4.79 (d, 3H), 4.59-4.55 (dd, 1H), 2.52-2.45 (m, 1H), 2.42-2.35 (q, 1H), 2.28-2.24 (d, 1H), 2.15-1.14 (t, 1H), 2.09 (s, 3H), 1.87-1.80 (m, 2H), 1.77-1.68 (m, 2H), 1.56-1.51 (m, 2H), 1.30 (s, 3H), 1.16-1.14 (d, 2H), 1.12 (s, 3H), 1.01 (s, 3H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ):177.5, 172.3, 171.0, 143.4, 138.1, 82.9, 79.2, 73.2, 70.5, 57.2, 53.3, 47.7, 39.1, 36.3, 36.1, 32.6, 31.3, 24.2, 23.8, 21.3, 19.6, 13.9.

[0455] Step 3: Synthesis of Compound 16

[0456] In DCM (15 mL) Intermediate 11 Oxalyl chloride (1.27 mL, 14.8 mmol, 3 eq.) was added to a solution of (2 g, 4.92 mmol, 1 eq.), then a catalytic amount of DMF (0.1 mL) was added at rt, and the resulting mixture was stirred at rt for an additional 3 h. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The obtained residue was dissolved in dry THF (12 mL) and added at 0°C to a mixture of 1-methylpiperazine (0.819 mL, 7.38 mmol, 1.5 eq.) and triethylamine (2.06 mL, 14.08 mmol, 3 eq.) in dry THF (3 mL), and the resulting mixture was stirred at rt for 1 h. The progress of the reaction was monitored by TLC. After the reaction was completed, water (80 mL) was added and extracted with EtOAc (4 x 40 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (100 to 200 mesh) using a mixture of MeOH in DCM as the eluent. Compound 16 was provided as a yellow solid. Yield: 1.5 g (62.39%).

[0457] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.28 (s, 1H), 4.82 (s, 2H), 4.74 (t, J=14.8 Hz, 1H), 3.65 (s, 4H), 3.22 (s, 1H), 2.82-2.74 (m, 1H), 2.53-2.48 (m, 1H), 2.42-2.38 (m, 4H), 2.30 (s, 3H), 2.08 (s, 5H), 1.86 (d, 1H), 1.73-1.70 (q, 2H), 1.66-1.57 (m, 3H), 1.40 (s, 3H), 1.20 (s, 3H), 1.13 (s, 3H), 1.08 (s, 3H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ): 173.0, 172.4, 171.2, 143.1, 138.3, 85.2, 82.2, 73.3, 70.4, 58.0, 55.4, 53.6, 50.1, 46.0, 45.7, 39.6, 36.5, 35.9, 33.0, 31.5, 26.9, 25.9, 24.8, 21.5, 20.2, 13.9.

[0458] Representative synthesis of Compound 17:

[0459]

[0460] Compound 17

[0461] DCM(8 mL) 중 Intermediate 11Oxalyl chloride (0.253 mL, 2.95 mmol, 3.0 eq.) was added to a solution of (400 mg, 0.984 mmol, 1 eq.), then a catalytic amount of DMF (0.02 mL) was added at rt, and the resulting mixture was stirred at rt for an additional 3 h. The progress of the reaction was monitored by TLC. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The obtained residue was dissolved in dry THF (6 mL) and added at 0°C to a mixture of morpholine (0.127 mL, 1.48 mmol, 1.5 eq.) and triethylamine (0.411 mL, 2.95 mmol, 3.0 eq.) in dry THF (2 mL), and the resulting mixture was stirred at rt for 1 h. The progress of the reaction was monitored by TLC. After the reaction was complete, water (80 mL) was added and extracted with EtOAc (3 x 30 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (100 to 200 mesh) using a mixture of MeOH in DCM as the eluent. Compound 17 It was provided as a yellow solid. Yield: 220 mg (47.01%).

[0462] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.28 (s, 1H), 4.82 (t, J=4.0 Hz, 2H), 4.74 (t, 1H), 4.59-4.55 (dd, 1H), 3.70-3.65 (m, 7H), 2.75-2.11 (m, 3H), 2.09 (s, 3H), 2.07-2.03 (m, 1H), 1.90-1.85 (m, 1H), 1.77-1.56 (m, 9H), 1.40 (s, 3H), 1.16 (s, 3H), 1.11 (s, 3H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ):173.2, 172.4, 171.1, 143.1, 138.3, 85.1, 82.1, 73.3, 70.4, 67.1, 58.0, 53.6, 50.1, 46.8, 39.6, 36.5, 35.9, 33.0, 31.5, 25.8, 24.8, 21.4, 20.2, 13.9.

[0463] Representative synthesis of compound 18:

[0464]

[0465] Compound 18

[0466]

[0467] (2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)-dodecahydronaphtho[2,1-b]furan-6-carbonitrile in DCM (15 mL) Compound 1 DMP (2.15 g, 5.07 mmol, 2.5 eq.) was added to a solution of (0.7 g, 2.03 mmol, 1 eq.) at rt. The mixture was stirred at rt for 3 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with DCM (80 mL) and washed with a saturated NaHCO3 solution (50 mL), a saturated hypochlorite solution (2 x 80 mL), and a saline solution (80 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product is purified by flash column chromatography using EtOAc in hexane (100 to 200 silica gel mesh) as the eluent to obtain the desired product (2S,3aR,5aS,6R,9aR,9bS)-3a,6,9a-trimethyl-7-oxo-2-(2-oxo-2,5-dihydrofuran-3-yl)-dodecahydronaphtho[2,1-b]furan-6-carbonitrile( Compound 18 ) was provided as a white solid. Yield: 510 mg (73.28%).

[0468] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.33 (d, J=1.6 Hz, 1H), 4.81 (t, J=15.6 Hz, 2H), 2.99-2.98 (dd, 1H), 2.50-2.10 (m, 2H), 2.47-2.01 (m, 4H), 1.73-1.59 (m, 6H), 1.51 (d, 5H), 1.44-1.27 (m, 3H), 1.16 (s, 3H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ): 202.4, 143.5, 137.9,119.7, 82.3, 73.2, 70.5, 56.4, 53.9, 46.5, 40.4, 36.0, 35.2, 34.5, 32.7, 31.3, 21.0, 20.6, 14.0.

[0469] Representative synthesis of compound 19:

[0470]

[0471] Compound 19

[0472]

[0473] A mixture of hydroxylamine hydrochloride (1.98 g, 28.5 mmol, 3.5 eq.) and pyridine (6.57 mL, 81.5 mmol, 10.0 eq.) in methanol (25 mL) of (2S,3aR,5aS, 6R,9aR,9bS)-3a,6,9a-trimethyl-7-oxo-2-(2-oxo-2,5-dihydrofuran-3-yl)-dodecahydronaphtho[2,1-b]furan-6-carbonitrile ( Compound 18)(2.8 g, 8.15 mmol, 1.0 eq.) was added dropwise at RT to a solution. The resulting mixture was stirred at RT for 3 h. The progress of the reaction was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated under reduced pressure, and the obtained crude product was washed with ice-cold water (200 mL). The precipitated solid was filtered and dried under vacuum to obtain the crude product (2S,3aR,5aS,6S,7E,9aR,9bS)-7-(hydroxyimino)-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)-dodecahydronaphtho[2,1-b]furan-6-carbonitrile Compound 19 It was provided as a grayish-white solid. The unrefined product was taken directly for the next step without further purification. Yield: 2.0 g (68.44%).

[0474] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.41 (s, 1H), 7.31 (q, 1H), 4.813-4.82 (q, 2H), 4.79-4.74 (m, 1H), 3.32-3.26 (m, 1H), 2.54-2.32 (q, 1H), 2.31-2.02 (m, 5H), 1.94-1.86 (m, 2H), 1.78-1.71 (m, 5H), 1.56 (s, 3H), 1.54-1.21 (m, 5H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ): 172.5, 157.3, 143.5, 137.8, 121.5, 82.4, 73.1, 70.5, 56.5, 52.4, 39.8, 38.4, 36.0, 34.4, 32.6, 31.2, 22.8, 20.1, 18.2, 13.8.

[0475] Representative synthesis of Compound 20:

[0476]

[0477] Compound 20

[0478]

[0479] (2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)-dodecahydronaphtho[2,1-b]furan-6-carbonitrile( Compound 1 Pd / C (200 mg) was added at rt to a solution of )(0.5 g, 1.45 mmol, 1 eq.). The resulting mixture was stirred at rt for 12 h under hydrogen gas pressure at rt. The reaction mixture was monitored by TLC. After the reaction was complete, the reaction mixture was filtered through a Celite layer, and the layer was washed with EtOH (20 mL). The combined filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography in hexane using EtOAC (100 to 200 silica gel mesh) as the eluent to obtain (2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxoxolane-3-yl)-dodecahydronaphth[2,1-b]furan-6-carbonitrile( Compound 20) Provided. Yield: 300 mg (59.65%).

[0480] 1 H NMR (400 MHz, DMSO, δ): 5.42 (d, 1H), 4.31-4.23 (m, 1H), 4.2-4.1 (m, 1H), 4.0-3.9 (m, 1H), 3.32 (s, 3H), 3.11 (m, 1H), 2.81-2.70 (m, 1H), 2.30-2.21 (m, 1H), 2.2-2.1 (m, 1H), 2.2-1.9 (m, 3H), 1.75-1.56 (m, 5H), 1.15-1.39 (m, 4H), 1.40-1.30 (s, 4H), 1.28-1.20 (m, 2H), 1.20-1.15 (m, 1H), 1.14 (s, 3H), 1.14-1.10 (m, 1H), 1.05 (s, 3H), 1.00 (m, 1H). 13C NMR (100 MHz, DMSO, δ):177.3, 122.7, 81.6, 76.1, 74.2. 70.5, 66.6, 57.3, 51.9, 46.1, 44.5., 43.2, 41.9, 38.4, 35.7, 32.5, 31.7, 30.9, 30.3, 29.5, 27.9. 24.4, 22.8, 19.2, 14.2.

[0481] Representative synthesis of compound 21:

[0482]

[0483] Compound 21

[0484]

[0485] 3-[(2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-6-(hydroxymethyl)-3a,6,9a-trimethyl-dodecahydronaphtho[2,1-b]furan-2-yl]-2,5-dihydrofuran-2-one in pyridine (2 mL) Isoandrographolide Acetic anhydride (4 mL) was added dropwise at rt to a solution of )(1 g, 2.85 mmol, 1 equivalent). The resulting mixture was maintained at rt for an additional 16 h. The reaction was monitored by TLC. After the reaction was complete, the reaction mixture was poured onto crushed ice (100 g), the solid was filtered, and dried under vacuum to yield a crude product (850 mg). The obtained crude product was purified by column chromatography on EtOAc (100 to 200 silica gel mesh) in hexane as the eluent to [(2S,3aR,5aS,6R,7R,9aR,9bS)-7-(acetyloxy)-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)-dodecahydronaphtho[2,1-b]furan-6-yl]methyl acetate ( Compound 21 ) was provided as a white solid. Yield: 0.34 g (27.42%).

[0486] 1 1H NMR (400 MHz, CDCl₃ 3 ): δ 7.28 (t, J=2.0Hz, 1H), 4.82 (s, 2H), 4.73 (t, J =9.2Hz, 1H), 4.59 (q, J =5.6Hz, 1H), 4.41 (d, J =11.6Hz, 1H), 4.20 (d, J =11.6Hz, 1H), 2.47 (q, J =8.0Hz, 1H), 2.22 (dd, J =11.2Hz, 3.2Hz, 1H), 2.05 (s, 6H), 1.82 (d, J =3.2Hz, 1H), 1.79 (d, J =3.2Hz, 1H), 1.71-1.62 (m, 5H), 1.58-1.42 (m, 2H), 1.28 (d, J =13.6Hz, 1H), 1.12 (s, 3H), 1.10 (d, J =4.4Hz, 1H), 1.06 (s, 3H), 1.03 (s, 3H). 13C NMR (100 MHz, CDCl 3 ): δ 172.5, 170.8, 170.5, 143.1, 138.4, 82.6, 80.12, 73.2, 70.5, 65.0, 57.8, 52.9, 40.9, 39.0, 36.4, 35.7, 32.8, 31.4, 23.5, 23.0, 21.1, 21.0, 18.8, 15.8.

[0487] Representative synthesis of compound 22:

[0488]

[0489] Compound 22

[0490]

[0491] Step 1: Synthesis of Intermediate 12

[0492] 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-6-(hydroxymethyl)-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one in DCM (150 mL) Isoandrographolide Triethylamine (7.56 mL, 54.2 mmol, 2 eq.) and trityl chloride (8.31 g, 29.8 mmol, 1.1 eq.) were added in fractions at rt to a solution of )(9.5 g, 27.1 mmol, 1 eq.). The resulting mixture was maintained at rt for an additional 16 h. The progress of the reaction mixture was monitored by TLC. After the completion of the reaction, the reaction mixture was diluted with DCM (150 mL) and washed with water (3 x 100 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (100 to 200 mesh) using a mixture of ethyl acetate and hexane as the eluent to obtain 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-6-((trityloxy)methyl)dodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one( Intermediate 12 ) was provided as a white solid. Yield: 1.2 g (7.47%).

[0493] 1 1H NMR (400 MHz, CDCl₃ 3 ): δ 7.47(t, J = 7.2 Hz, 5H), 7.34 (t, J = 7.2 Hz, 5H), 7.27(t, J = 4.8 Hz, 5H), 7.24(t, J = 2.0 Hz, 1H), 4.79(t, J = 2.0 Hz, 1H), 4.57 (t, J = 7.6 Hz, 1H), 4.14 (q, J = 7.2 Hz, 1H), 3.47(d, J = 9.6 Hz, 1H), 3.20 (dd, J = 22.0, 9.2 Hz, 1H ), 2.31(dd, J = 13.6, 7.6 Hz, 8.0 Hz, 2H), 2.21(dd,J = 10.0, 5.2 Hz, 2H), 1.96-1.88(m, 2H), 1.75-1.67(m, 2H), 1.55 (d, J = 11.2 Hz, 1H), 1.53(s, 3H), 1.46(t, J = 8.0 Hz, 2H), 1.28(t, J = 7.2 Hz, 2H), 1.08 ( s, 3H), 1.04-0.90 ( m, 3H). 13C NMR (100 MHz, CDCl 3 ): δ 172.5, 143.4, 143.0, 138.3, 128.5, 128.0, 127.2, 87.7, 82.6, 80.6, 73.0, 70.4, 64.8, 60.4, 52.7, 42.4, 39.3, 35.8, 35.6, 33.0, 31.5, 27.4, 23.3, 18.2, 16.1, 14.2.

[0494] Step 2: Synthesis of Intermediate 13

[0495] In pyridine (3 mL), 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-6-((trityloxy)methyl)dodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one( Intermediate 12Acetic acid solvent (90.4 mg, 0.886 mmol, 1.5 eq.) was added at rt to a solution of )(350 mg, 0.590 mmol, 1 eq.). The resulting mixture was stirred at rt for an additional 3 h. The progress of the reaction mixture was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with DCM (50 mL) and washed with water (3 x 20 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (100 to 200 mesh) using a mixture of ethyl acetate and hexane as the eluent to obtain (2S,3aR,5aS,6R,7R,9aR,9bS)-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)-6-((trityloxy)methyl)dodecahydronaphtho[2,1-b]furan-7-yl acetate ( Intermediate 13 ) was provided as a grayish-white solid. Yield: 0.3 g (80.04%).

[0496] 1 1H NMR (400 MHz, CDCl₃ 3 ): δ 7.52-7.49 (m, 5H), 7.32-7.24 (m, 10H), 7.23 (t, J =2.4Hz, 1H), 4.81 (t, J =2.0Hz, 2H), 4.62 (t, J =7.6Hz, 1H), 4.56 (dd, J =12Hz, 4.0Hz, 12.0Hz, 1H), 3.42 (d, J =9.6Hz, 1H), 3.15 (d, J =9.6Hz, 1H), 2.36 (q, J =9.0Hz, 1H), 2.10 (dt, J =13.2Hz, 4.4Hz, 1H), 2.04 (s, 3H), 2.00-1.93 (m, 2H), 1.67 (dt, J =13.6Hz, 3.6Hz, 1H), 1.64 (dd, J=13.6Hz, 3.6Hz, 1H), 1.50 (d, J =7.6Hz, 2H), 1.46-1.39 (m, 2H), 1.28 (s, 3H), 1.18 (s, 3H), 1.10 (s, 3H), 0.92-0.85 (m, 2H), 0.61 (s, 3H). 13C NMR (100 MHz, CDCl 3 ): δ 172.5, 170.8, 144.2, 143.0, 138.4, 128.8, 127.6, 126.8, 87.0, 82.7, 80.6, 73.1, 70.5, 62.6, 58.1, 52.8, 42.2, 39.2, 36.1, 35.6, 32.9, 31.5, 29.7, 23.6, 23.3, 21.3, 18.9, 15.5.

[0497] Step 3: Synthesis of Compound 22

[0498] (2S,3aR,5aS,6R,7R,9aR,9bS)-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)-6-((trityloxy)methyl)dodecahydronaphtho[2,1-b]furan-7-yl acetate in DCM(5 mL) Intermediate 13Formic acid (5 mL) was added dropwise at rt to a solution of )(280 mg, 0.441 mmol, 1 eq.). The resulting mixture was stirred at rt for an additional 7 h. The progress of the reaction mixture was monitored by TLC. After the reaction was complete, the reaction mixture was basicized with a saturated NaHCO3 (15 mL) solution. The organic layer was separated, and the aqueous layer was extracted with DCM (2 x 20 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (100 to 200 mesh) using a mixture of ethyl acetate and hexane as the eluent to obtain (2S,3aR,5aS,6R,7R,9aR,9bS)-6-((formyloxy)methyl)-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-7-yl acetate ( Compound 22 ) was provided as a grayish-white solid. transference number: 0.14 g (75.48%).

[0499] 1 1H NMR (400 MHz, CDCl₃ 3 ): δ 8.08( d, J = 8.0 Hz, 1H), 4.82(s, 2H), 4.72 (t, J = 7.6 Hz, 1H), 4.62-4.54 (m, 2H), 4.40 (d, J = 11.6 Hz, 1H), 4,23(t, J = 11.6 Hz, 1H), 2.46( q, J = 8.0 Hz, 1H), 2. 22(d, 14.4 Hz, 1H), 2.09(t, J = 4.4 Hz, 2H), 2.04(s, 3H), 1.82-1.51( m, 8H), 1.27( d, 7.6 Hz, 1H), 1.12(s, 3H), 1.08(s, 3H), 1.03(s, 3H). 13C NMR (100 MHz, CDCl 3 ):δ 172.5, 170.5, 161.0, 143.1, 138.4, 82.5, 79.9, 73.2, 70.5, 64.2, 57.8, 52.8, 40.9, 38.9, 36.2, 35.7, 32.8, 31.4, 23.5, 22.9, 21.1, 18.7, 15.8.

[0500] Representative synthesis of Compound 23:

[0501]

[0502] Compound 23

[0503]

[0504] (3-[(2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-6-(hydroxymethyl)-3a,6,9a-trimethyl-dodecahydronaphtho[2,1-b]furan-2-yl]-2,5-dihydrofuran-2-one) in 2,2-dimethoxypropane (2.5 mL) Isoandrographolide A catalytic amount of camphorsulfonic acid (16.6 mg, 0.0713 mmol, 0.05 eq.) was added to a solution of )(500 mg, 1.43 mmol, 1 eq.) at rt. The resulting mixture was maintained at rt for an additional 5 h. The reaction was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated under reduced pressure, the residue was diluted with EtOAc (30 mL), and washed with a saturated NaHCO3 solution (15 mL) and water (15 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to provide a crude product. The obtained crude product (100 to 200 silica gel mesh) was purified by column chromatography using EtOAc:hexane as the eluent (3-[(1S,2R,7R,10R,11S,13S,15R)-2,5,5,10,15-pentamethyl-4,6,14-trioxatetracyclo[8.7.0.0 2 , 7 .0 11 , 15 ]heptadecan-13-yl]-2,5-dihydrofuran-2-one)( Compound 23) was provided as a white solid. Yield: 0.33 g (59.23%)

[0505] 1 1H NMR (400 MHz, CDCl₃ 3 ): δ 7.29(t, J = 1.6 Hz, 1H), 4.82(d, J = 1.6 Hz, 2H), 4.74 (t, J = 9.2 Hz, 1H), 4.10 (d, J = 12 Hz, 1H), 3.48 (dd, J = 10.4, 4.0 Hz, 1H), 3.27(d, J = 12.0 Hz, 1H), 2.50 (q, J = 8.0 Hz, 1H), 2.21(dt, J = 13.2, 4.4 Hz, 2H), 2.08-1.99( m, 2H), 1.59(d, 7.6 Hz, 1H), 1.54 (t, J= 8.4 Hz, 1H), 1.48(t, J= 8.0 Hz, 2H), 1.46(s, 3H), 1.39( s, 3H), 1.25( s, 3H), 1.17(s, 3H), 1.12 ( s, 3H), 1.07-0.95(m, 3H). 13C NMR (100 MHz, CDCl 3 ): δ 172.5, 143.1, 138.5, 98.7, 82.8, 77.6, 73.1, 70.5, 63.7, 57.9, 50.5, 37.1, 37.0, 36.0, 35.1, 32.8, 31.6, 27.7, 26.6, 25.9, 25.1, 17.5, 17.2.

[0506] Representative synthesis of Compound 24:

[0507]

[0508] Compound 24

[0509]

[0510] Step 1: Synthesis of Intermediate 14

[0511] 3-{(1R,2S,4S,6R,9S,10R,11R)-11-hydroxy-10-(hydroxymethyl)-1,6,10-trimethyl-5-oxatricyclo[7.4.0.0 in DCM(10 mL) 2 , 6 ]Tridek-4-il}-2(5H)-Furanon( Isoandrographolide PCC (615 mg, 2.85 mmol, 2 eq.) was added at rt to a solution of )(500 mg, 1.43 mmol, 1 eq.). The resulting mixture was stirred at rt for an additional 3 h. The reaction was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated under reduced pressure. The obtained crude product was purified by column chromatography using EtOAc (100 to 200 silica gel mesh) in hexane as the eluent, and (((1R,2S,4S,6R,9S,10R)-1,6,10-trimethyl-11-oxo-4-(2-oxo-3-furyl)-5-oxatricyclo[7.4.0.0 2 , 6 ]tridecane-10-carbaldehyde)( Intermediate 14 ) was provided as a grayish-white solid. Yield: 140 mg, (28.33%).

[0512] 1 1H NMR (400 MHz, CDCl₃ 3 ) δ 9.70 (s, 1H), 7.30 (d, J = 1.6Hz, 1H), 4.83(d, J = 6.0 Hz, 2H), 2.67(td, J = 14.8, 8.8 Hz, 1H), 2.54(q, J = 8.0 Hz, 1H), 2.42(dt, J = 14.2, 2.8 Hz, 1H), 2.33(dt, J = 14.2, 2.8, 1H), 2.18-2.13(m, 1H), 2.11-2.03(m, 1H), 1.96(td, J = 12.8, 4.4 Hz, 1H), 1.80(dt, J= 13.2, 2.8 Hz, 2H), 1.64(d, J = 7.6 Hz, 1H), 1.58(q, J = 7.6 Hz, 1H), 1.50(dd, J = 12.8, 2.0 Hz, 1H), 1.38(td, J = 18.4, 4.4 Hz, 1H), 1.31(s, 3H), 1.17(s, 3H), 1.15(s, 3H). 13C NMR (100 MHz, CDCl 3 ) δ 208.5, 201.3, 172.4, 143.2, 138.2, 82.3, 73.2, 70.5, 63.3, 55.7, 55.3, 39.6, 36.2, 36.1, 35.9, 32.9, 31.1, 19.2, 17.2, 14.8.

[0513] Step 2: Synthesis of Compound 24

[0514] ((2S,3aR,5aS,6R,9aR,9bS)-3a,6,9a-trimethyl-7-oxo-2-(2-oxo-2,5-dihydrofuran-3-yl)-dodecahydronaphtho[2,1-b]furan-6-carbaldehyde)( Intermediate 14 Hydrazine hydrate (21.7 mg, 0.433 mmol, 1.5 eq.) was added at rt to a solution of )(100 mg, 0.289 mmol, 1 eq.). The resulting mixture was stirred at rt for an additional 3 h. The reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with water (20 mL) and extracted with DCM (2 x 30 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography using MeOH (100 to 200 silica gel mesh) in DCM as the eluent to obtain 3-[(1S,2R,9R,10S,12S,14R)-2,9,14-trimethyl-13-oxa-4,5-diazatetracyclo[7.7.0.0 2 , 6 .0 10 , 14]hexadeca-3,5-dien-12-yl]-2,5-dihydrofuran-2-one( Compound 24 ) was provided as a white solid. Yield: 50 mg, (50.58%).

[0515] 1 1H NMR (400 MHz, CDCl₃ 3 ): δ 8.05 (s, 1H), 4.82 (d, J =6.0Hz, 2H), 4.68 (t, J =8.4Hz, 1H), 3.08-2.99 (m, 1H), 2.82 (q, J =10.4Hz, 1H), 2.48 (q, J =8.4Hz, 1H), 2.10 (dd, J =8.8Hz, 2.8Hz, 1H), 2.04-1.98 (m, 3H), 1.88-1.80 (m, 1H), 1.7 (dd, J =12.4Hz, 2.0Hz, 2H), 1.63-1.52 (m, 3H), 1.31 (s, 3H), 1.13 (s, 3H), 0.59 (s, 3H). 13C NMR (100 MHz, CDCl 3 ): δ 172.4, 170.9, 143.2, 138.3, 82.4, 70.5, 64.4, 54.8, 54.7, 37.1, 36.5, 35.5, 33.1, 31.6, 23.7, 22.7, 20.7, 13.5.

[0516] Representative synthesis of Compound 25:

[0517]

[0518] Compound 25

[0519]

[0520] Step 1: Synthesis of Intermediate 15

[0521] ((3E,4S)-3-{2-[(1R,4aS,5R,6R,8aS)-6-hydroxy-5-(hydroxymethyl)-5,8a-dimethyl-2-methylidene-decahydronaphthalene-1-yl]ethylidene}-4-hydroxyoxolan-2-one)( Andrographolide )(50 g, 143 mmol, 1 eq.) and concentrated HCl (250 mL, 5 volumes) were mixed while stirring until the solution was completely dissolved. The resulting mixture was left aside for 48 h. The reaction was monitored by TLC. After the reaction was complete, the reaction mixture was slowly poured onto crushed ice, and the precipitated or separated solid was stirred at rt for 2 h. The solid was filtered, washed with water (2000 mL), and dried under vacuum. The filtrate is concentrated under reduced pressure, and the residue is purified by column chromatography on silica gel (100 to 200 mesh) using a mixture of acetone in DCM as the eluent ([(2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)-dodecahydronaphtho[2,1-b]furan-6-yl]methyl acetate)( Intermediate 15 ) provided. Yield: 1.9 g, 3.39%.

[0522] 1 1H NMR (400 MHz, CDCl₃ 3 ): δ 7.28(t, J = 1.6 Hz, 1H), 4.81(s, 2H), 4.71(t, J = 9.2 Hz, 1H), 4.39 (d, J = 11.6 Hz, 1H), 4.18(d, J = 11.6 Hz, 1H), 3.30(dd, J = 10.8, 4.8 Hz, 1H), 2.46 (q, J = 8.0 Hz, 1H), 2.21(dd, J = 13.2, 3.2 Hz, 1H), 2.06(s, 3H), 2.01(dd,J = 8.0, 3.2 Hz, 2H), 1.79(dd, J = 13.2, 3,2 Hz, 1H), 1.76-1.67(m, 2H), 1.61(q, J = 7.2 Hz, 2H), 1.55( q, J = 5.6 Hz, 2H), 1.17(s, 3H), 1.12(s, 3H), 1.03(dd, J = 16.8, 3.6 Hz, 2H), 0.98(s, 3H). 13C NMR (100 MHz, CDCl 3 ): δ 172.5, 171.1, 143.2, 138.4, 82.6, 79.2, 73.2, 70.5, 65.1, 57.9, 52.9, 42.1, 39.3, 36.3, 35.8, 32.8, 31.4, 27.0, 22.7, 21.1, 18.5, 16.0.

[0523] Step 2: Synthesis of Compound 25

[0524] (in DCM (10 mL) [(2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)-dodecahydronaphtho[2,1-b]furan-6-yl]methyl acetate) Intermediate 15 PCC (275 mg, 1.27 mmol, 2 eq.) was added at rt to a solution of )(250 mg, 0.637 mmol, 1 eq.). The resulting mixture was stirred at rt for an additional 3 h. The reaction was monitored by TLC. After the reaction was complete, the reaction mixture was concentrated under reduced pressure. The obtained crude product was purified by column chromatography in hexane (100 to 200 silica gel mesh) using EtOAc as the eluent to obtain ([(2S,3aR,5aS,6R,9aR,9bS)-3a,6,9a-trimethyl-7-oxo-2-(2-oxo-2,5-dihydrofuran-3-yl)-dodecahydronaphtho[2,1-b]furan-6-yl]methyl acetate)( Compound 25) was provided as a white solid. Yield: 170 mg, (68.35%).

[0525] 1 1H NMR (400 MHz, CDCl₃ 3 ): δ 7.31(t, J = 1.6 Hz, 1H), 4.84(d, J = 1.6 Hz, 2H), 4.78 (t, J = 9.2 Hz, 1H), 4.62 (d, J = 11.2 Hz, 1H), 4.99(d, J = 11.6 Hz, 1H), 2.79(td, J = 9.2, 6.0 Hz, 1H), 2.54 (q, J = 8.0 Hz, 1H), 2.34(dt, J = 13.6, 1.6 Hz, 1H), 2.27 (dd, J = 12.0, 6.2 Hz, 1H), 2.13- 2.03(m, 2H), 2.01(s, 3H), 1.65-154 (m, 4H), 1.43- 1.48(m, 2H), 1.27 (s, 3H), 1.18 (s, 3H), 1.14 (s, 3H). 13C NMR (100 MHz, CDCl 3 ): δ 212.6, 172.4, 170.7, 143.2, 138.3, 82.5, 73.1, 70.5, 66.2, 57.0, 54.5, 51.4, 39.4, 36.0, 35.5, 34.9, 32.9, 31.4, 21.3, 20.7, 19.2, 15.6.

[0526] Representative synthesis of compound 26:

[0527]

[0528] Compound 26

[0529]

[0530] 3-((2S,3aR,5aS,6R,7R,9aR,9bS)-7-hydroxy-6-(hydroxymethyl)-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one in THF(5 mL) Isoandrographolide Pyridine (0.069 mL, 0.856 mmol, 3 eq.) was added at rt to a solution of )(100 mg, 0.285 mmol, 1 eq.). The reaction mixture was cooled to 0°C, and then triphosgene (127 mg, 0.428 mmol, 1.5 eq.) was added in portions. The resulting mixture was stirred at 0°C for 1 h. The reaction mixture was quenched with a saturated NaHCO3 solution (20 mL) and extracted with ethyl acetate (3 x 25 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography using acetone in DCM (100 to 200 silica gel mesh) as the eluent to obtain (4aR,4bS,6aR,8S,9aS,9bR,11aR)-4a,6a,9b-trimethyl-8-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydro-4H-furo[3',2':5,6]naphtho[2,1-d][1,3]dioxin-2-one( Compound 26 ) was provided as a grayish-white solid. Yield: 60 mg (55.86%).

[0531] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.29 (d, J =6.4 Hz, 1H), 4.83-4.68 (m, 4H), 4.08 (t, 1H), 3.90 (d, J = 11.2 Hz, 1H), 2.48-2.43 (q, 1H), 2.27-2.23 (q, 1H), 2.10-2.04 (m, 2H), 1.88-1.81 (q, 4H), 1.46-1.44 (m, 1H), 1.32 (s, 3H), 1.27 (d, 1H), 1.20-1.10 (m, 4H), 1.08-1.03 (m, 5H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ): 172.4, 148.3, 143.3, 137.9, 73.0, 58.0, 50.5, 37.2, 35.5, 35.4, 34.7, 32.7, 31.6, 25.4, 23.9, 17.8, 16.3.

[0532] Representative synthesis of Compound 27:

[0533]

[0534] Compound 27

[0535]

[0536] (2S,3aR,5aS,6S,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbaldehyde in ACN (20 mL) Intermediate 10 Ag2O (3.35 g, 14.3 mmol, 5 eq.), CaSO4 (2.34 g, 17.2 mmol, 6 eq.), and CH3I (8.15 g, 57.4 mmol, 20 eq.) were added at rt to a solution of )(1 g, 2.87 mmol, 1 eq.). The resulting mixture was stirred at rt for 30 h, and then heated at 60°C for an additional 12 h. The reaction mixture was monitored by TLC. After the reaction was complete, the reaction mixture was filtered through a silica gel pad and washed with ethyl acetate (100 mL). The combined filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography in hexane (100 to 200 silica gel mesh) using EtOAc as the eluent to obtain (2S,3aR,5aS,6S,7R,9aR,9bS)-7-methoxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carboxylic acid ( Compound 27 ) was provided as a grayish-white solid. Yield: 350 mg (32.22%).

[0537] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.28 (s, 1H), 4.81 (s, 2H), 4.72 (brs, 1H), 3.67 (s, 3H), 3.39 (d, J =12 Hz, 1H), 3.11 (t, 1H), 2.50-2.45 (q, 1H), 2.22 (d, J=14.8 Hz, 1H), 2.09-1.99 (m, 2H), 1.79 (s, 5H), 1.54 (d, J=7.2 Hz, 1H), 1.43 (s, 3H), 1.39 (d, 1H), 1.10 (s, 4H), 1.04 (d, 1H), 0.84 (s, 3H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ): 178.2, 172.5, 143.1, 138.5, 82.5, 78.4, 73.3, 70.5, 56.8, 53.8, 51.3, 49.0, 39.9, 36.6, 36.1, 32.9, 31.2, 27.9, 23.9, 19.7, 14.2.

[0538] Representative synthesis of compound 28:

[0539]

[0540] Compound 28

[0541]

[0542] 3-((2S,3aR,5aS,6S,9aR,9bS,E)-6-(((tert-butyldimethylsilyl)oxy)methyl)-7-(hydroxyimino)-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one in DCM (10 mL) Intermediate 54N HCl (2.61 mL, 5 eq.) in a dioxane solution was added to a solution of )(1 g, 2.09 mmol, 1 eq.) at 0°C. The resulting reaction mixture was stirred at rt for 2 h. The reaction was monitored by TLC. After the reaction was complete, the reaction mixture was diluted with DCM (50 mL) and basicized (P) with a saturated NaHCO3 solution (50 mL). H =8) and then extracted with DCM (3 x 40 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain 3-((2S,3aR,5aS,6S,9aR,9bS,E)-7-(hydroxyimino)-6-(hydroxymethyl)-3a,6,9a-trimethyldodecahydronaphtho[2,1-b]furan-2-yl)furan-2(5H)-one( Compound 28 ) was provided as a white solid. Yield: 650 mg (85.43%).

[0543] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.85 (brs, 1H), 7.30 (d, J =2.0 Hz, 1H), 4.83 (d, J=1.6 Hz, 2H), 4.75 (t, 1H), 3.84 (d, J=10.8 Hz, 1H), 3.48 (d, J =8.4 Hz, 1H), 3.05-2.98 (dd, 1H), 2.76 (brs, 1H), 2.54-2.49 (q, 1H), 2.32-2.23 (m, 2H), 2.13-2.07 (m, 1H), 1.57-1.53 ​​(m, 1H), 1.36-1.32 (t, 1H), 1.27 (s, 3H), 1.22-1.15 (m, 1H), 1.13 (s, 3H), 1.05 (s, 3H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ):172.5, 164.7, 143.1, 138.5, 82.5, 73.1, 70.5, 65.4, 56.7, 53.0, 44.6, 36.9, 36.2, 35.4, 32.9, 31.4, 23.6, 18.5, 18.0, 15.5.

[0544] Representative synthesis of compound 29:

[0545]

[0546] Compound 29

[0547]

[0548] (E)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxo-2,5-dihydrofuran-3-yl)dodecahydronaphtho[2,1-b]furan-6-carbaldehyde oxime in THF (10 ml) Intermediate 2 SOCl2 (239 μL, 3.3 mmol, 1.5 eq.) was added dropwise to a solution of )(800 mg, 2.2 mmol, 1.0 eq.) at 0°C. The resulting reaction mixture was stirred at 0°C for 30 min. The reaction was monitored by TLC. After the reaction was complete, a saturated NaHCO3 solution was added to the reaction mixture, extracted with ethyl acetate (2 x 20 mL), the combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude material was purified by column chromatography using silica (100 to 120 mesh) and ethyl acetate in hexane as the eluent to obtain 3-((3aR,3bS,5aR,7S,8aS,8bR,10aR)-3a,5a,8b-trimethyl-3a,3b,4,5,5a,7,8,8a,8b,9,10,10a-dodecahydrofuro[3',2':5,6]naphtho[1,2-d]isoxazole-7-yl)furan-2(5H)-one( Compound 29 ) was provided as a grayish-white solid. Yield: 70 mg (9.21%).

[0549] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 7.26 (d,J =5.2 Hz, 1H), 7.12 (s, 1H), 4.80 (s, 2H), 4.71 (t, 1H), 4.15-4.11 (q, 1H), 2.49-2.43 (q, 1H), 2.27 (d, J=10.8 Hz, 1H), 2.08-2.00 (m, 1H), 1.84 (m, 1H), 1.61-1.42 (m, 7H), 1.20 (m, 1H), 1.15 (s, 3H), 1.11 (s, 3H), 0.93 (s, 3H), 0.87 (d, 1H). 13 13C NMR (100 MHz, CDCl₃ 3 , δ): 172.4, 153.6, 143.2, 138.4, 84.2. 82.6, 73.1, 70.5, 56.0, 53.6, 48.7, 35.8, 35.1, 34.8, 32.5, 31.5, 25.1, 24.2, 19.9, 14.2.

[0550] Representative synthesis of Compound 30:

[0551]

[0552] Compound 30

[0553]

[0554] 3-{(1R,2S,4S,6R,9S,10R,11R)-11-hydroxy-10-(hydroxymethyl)-1,6,10-trimethyl-5-oxatricyclo[7.4.0.0 in methanol (80 mL) 2 , 6Pd / C (1 g) was added at rt to a solution of ]tridec-4-yl}-2(5H)-furanone(isoandrographolide) (2 g, 5.71 mmol, 1 eq.). The resulting mixture was stirred at rt for 12 h under hydrogen gas pressure at rt. The reaction mixture was monitored by TLC. After the reaction was complete, the reaction mixture was filtered through a Celite layer, and the layer was washed with MeOH (100 mL). The combined filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography using MeOH in DCM (100 to 200 silica gel mesh) as the eluent to obtain the desired product 3-{(1R,2S,6R,9S,10R,11R)-11-hydroxy-10-(hydroxymethyl)-1,6,10-trimethyl-5-oxatricyclo[7.4.0.0 2 , 6 ]tridec-4-yl}-4,5-dihydro-2(3H)-furanone (compound 30) was provided. Yield: 850 mg (42.26%).

[0555] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 5.04 (d, J = 6.0 Hz, 1H), 4.3-4.2 (m, 1H), 4.18-4.10 (m, 2H), 3.95-3.71 (m, 2H), 3.21-3.19 (m, 1H), 2.78-2.71 (m, 1H), 2.32-2.24 (m, 1H), 2.14-1,82 (m, 4H), 1.62-1.41 (m, 6H), 1.10 (s, 3H), 1.06 (s, 3H), 1.02-0.91 (m, 2H), 0.83 (s, 3H).

[0556] Representative synthesis of Compound 31:

[0557]

[0558] Compound 31

[0559]

[0560] 3-{(1R,2S,6R,9S,10R,11R)-11-hydroxy-10-(hydroxymethyl)-1,6,10-trimethyl-5-oxatricyclo[7.4.0.0 in DCM(10 mL) 2 , 6 Solutions of TEMPO (35.5 mg, 0.2227 mmol, 0.2 eq.), TBAI (83.8 mg, 0.227 mmol, 0.2 eq.), NCS (227 mg, 1.7 mmol, 1.5 eq.), 0.5 M aqueous NaHCO3 (5 mL), and 0.05 M aqueous K2CO3 (5 mL) were added to a solution of ]tridec-4-yl-4,5-dihydro-2(3H)-furanone (compound 30) (0.4 g, 1.13 mmol, 1 eq.) at 0°C. The resulting mixture was stirred at rt for 2 h. The reaction mixture was monitored by TLC. After the reaction was complete, the reaction mixture was quenched with water (50 ml), extracted with DCM (3 x 30 mL), and the organic layer was washed with a saturated sodium thiosulfate solution (3 x 40 mL). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure. The obtained crude product was purified by column chromatography in hexane using EtOAc (100 to 200 silica gel mesh) as the eluent to obtain (2S,3aR,5aS,6S,7R,9aR,9bS)-7-hydroxy-3a,6,9a-trimethyl-2-(2-oxoxolane-3-yl)-dodecahydronaphtho[2,1-b]furan-6-carbaldehyde( Compound 31 ) was provided as a yellow solid. Yield: 300 mg (75.43%).

[0561] 1 1H NMR (400 MHz, CDCl₃ 3 , δ): 9.92 (s, 1H), 5.03= (d, J = 4.0 Hz, 1H), 4.27 (dt, J1 = 2.4 Hz, J2 = 8.4 Hz, 1H ), 4.13 (dd, J1 = 6.8 Hz, J2= 9.2 Hz, 1H), 3.76 (brs, 1H), 3.32-3.25 (m, 1H), 2.62-2.53 (m, 1H), 2-41-2.32 (m, 1H), 1.98-1.82 (m, 2H), 1.76-1.52 (m, 4H), 1.48-1.44 (m, 2H), 1.47-1.40 (m, 4H), 1.10-1.01 (m, 6H), 0.45 (s, 3H), 0.23-0.26 (m, 1H).

[0562] Example 2: Effect of Compound II on Immune Cell Populations in Ovalbumin-Induced Inflammation Model

[0563] The effects of Compound II on immune cell populations were investigated using an ovalbumin-induced inflammation model in female BALB / c mice. As shown in Figure 2, the experimental protocol involved sensitization with ovalbumin followed by treatment with dexamethasone or various doses of Compound II (0.1, 1, 10, and 30 mg / kg BID). This study aimed to evaluate the effects of Compound II on lymphocyte, eosinophil, and neutrophil counts in comparison to a control group.

[0564] The results demonstrated that treatment with Compound II resulted in a dose-dependent reduction in the number of lymphocytes, eosinophils, and neutrophils. At the maximum dose of 30 mg / kg BID, Compound II showed efficacy similar to dexamethasone in reducing these immune cell populations. These results suggest that Compound II can regulate several immune cell types known to express Rac2, including T cells and neutrophils, thereby supporting its potential as a broad-spectrum anti-inflammatory agent.

[0565] Example 3: Effect of Compound II on Body Weight and White Blood Cell Count

[0566] To evaluate the safety and systemic effects of Compound II, changes in body weight and white blood cell counts were monitored in an ovalbumin-induced inflammation model. Figures 3a, 3b, and 3c illustrate the study design and results for measurements of body weight, WBC / white blood cell count, and monocyte count.

[0567] Treatment with Compound II did not have a significant effect on body weight across all tested doses, suggesting good tolerability. WBC / white blood cell count and monocyte count showed a dose-dependent decrease with treatment with Compound II, and the highest dose (30 mg / kg BID) demonstrated an effect similar to dexamethasone. These results indicate that Compound II can regulate various white blood cell populations, including monocytes known to express Rac2, without causing significant adverse effects on body weight.

[0568] Example 4: Effect of Compound II on Serum IgE and Eotaxin Levels

[0569] The effect of Compound II on key inflammatory markers was evaluated by measuring serum IgE and eotaxin (BALF) concentrations in an ovalbumin-induced inflammation model. Figures 4a and 4b present the results of this analysis comparing the effects of Compound II at various doses to the control group and dexamethasone treatment.

[0570] Compound II demonstrated a dose-dependent reduction in both serum IgE and eotaxin levels. At the peak dose of 30 mg / kg BID, Compound II showed efficacy similar to dexamethasone in reducing these inflammatory markers. The reduction in serum IgE levels suggests that Compound II may affect B cells, which are known to express Rac2 and play a crucial role in IgE production. These results support the potential of Compound II as a therapeutic agent for allergic and inflammatory conditions.

[0571] Example 5: Effects of Compound II on MCP-1 Levels and Peribronchial Inflammation

[0572] To further characterize the anti-inflammatory properties of Compound II, its effects on MCP-1 (CCL2) levels in lung tissue and peribronchial inflammation were evaluated. Figures 5a and 5b illustrate the results of this analysis in an ovalbumin-induced inflammation model.

[0573] Treatment with Compound II resulted in a dose-dependent reduction in both MCP-1 levels and peribronchial inflammation scores. The maximum dose of Compound II (30 mg / kg BID) demonstrated efficacy similar to dexamethasone in reducing these inflammatory markers. These results suggest that Compound II may regulate macrophage function, as MCP-1 is a key chemokine involved in macrophage recruitment and activation. These findings are consistent with the known expression of Rac2 in macrophages and support the broad anti-inflammatory effects of Compound II.

[0574] Example 6: Transcriptive analysis of the effect of Compound II in an atopic dermatitis model

[0575] To elucidate the molecular mechanism underlying the therapeutic effects of Compound II, a comprehensive transcriptional analysis was performed using both human atopic dermatitis (AD) samples and an MC-903 mouse model. Figure 6 presents a heatmap visualization comparing gene expression patterns across different experimental conditions.

[0576] The analysis revealed that treatment with Compound II reduced the expression of genes upregulated in AD and restored the expression of genes downregulated in the disease state. In particular, pathway analysis demonstrated benefits in inflammatory cytokine signaling and barrier function. These results suggest that Compound II can modulate various aspects of AD pathophysiology through its effects on various Rac2-expressing cell types potentially involved in the disease process.

[0577] Example 7: Efficacy of Compound II in Reduction of Dermatitis Severity in MC-903 Mouse Model

[0578] The therapeutic potential of Compound II in treating atopic dermatitis was evaluated using an MC-903 mouse model. Figures 7a and 7b illustrate the study design and results, tracking dermatitis severity scores over time and presenting representative photographs of the back skin of the mice.

[0579] Treatment with Compound II resulted in a dose-dependent reduction in dermatitis severity scores, and higher doses demonstrated efficacy similar to the topical corticosteroid clobetasol. Gross examination of the skin in mice confirmed these results, demonstrating significant improvement in skin appearance with Compound II treatment. These findings support the potential of Compound II as a therapeutic agent for atopic dermatitis, which is likely achieved through the modulation of Rac2 activity in various cell types involved in the disease process.

[0580] Example 8: Dose-response analysis of Compound II in MC-903 atopic dermatitis model

[0581] A comprehensive dose-response analysis of Compound II was performed using the MC-903 atopic dermatitis model. Figures 8a and 8b present the study design and results, plotting the dermatitis severity score over time and the dose-response curve for Compound II.

[0582] Compound II demonstrated a clear dose-dependent effect on the severity of dermatitis, and higher doses (10 and 30 mg / kg BID) showed efficacy similar to clobetasol. The dose-response curves revealed a well-defined therapeutic range supporting the potential for once-daily administration in future clinical applications. These results further validate the therapeutic potential of Compound II in treating atopic dermatitis through the modulation of Rac2 activity.

[0583] Example 9: Comparative analysis of Compound II with standard therapy

[0584] To evaluate the relative efficacy of Compound II, a comparative analysis was performed against standard therapies including clobetasol and upadacitinib. Figure 9 presents a bar graph showing the fold change in response to different treatments in the MC-903 atopic dermatitis model.

[0585] Compound II demonstrated dose-dependent efficacy, and higher doses (10 and 30 mg / kg BID) showed effects similar to or superior to clobetasol and upadacitinib in reducing inflammatory markers. These results suggest that Compound II may provide a novel therapeutic approach for atopic dermatitis, potentially offering advantages over existing treatments through the specific modulation of Rac2 activity.

[0586] Example 10: Histological analysis of the effect of Compound II on epidermal thickness

[0587] To evaluate the effect of Compound II on skin histology, epidermal thickness measurements were performed in the MC-903 atopic dermatitis model. Figures 10a and 10b present microscopic images of skin sections and a quantitative analysis of epidermal thickness across different treatment conditions.

[0588] Treatment with Compound II resulted in a dose-dependent reduction in epidermal thickness, and higher doses demonstrated effects similar to clobetasol. These results suggest that Compound II can regulate keratinocyte function and proliferation, potentially through its effect on Rac2 activity in these cells. The ability to normalize skin histology supports the therapeutic potential of Compound II in treating atopic dermatitis.

[0589] Example 11: Analysis of Rac2 expression in atopic dermatitis

[0590] To establish the validity of Rac2 as a therapeutic target, Rac2 expression levels were analyzed across different experimental conditions and disease states. Figures 11a, 11b, and 11c present box plots illustrating the correlation between Rac2 expression and disease severity in mouse models and human AD samples.

[0591] Rac2 expression was significantly upregulated in both mouse and human AD samples compared to the control group. Furthermore, Rac2 expression levels were positively correlated with disease severity in human AD patients. Treatment with Compound II reduced Rac2 expression in the mouse model, suggesting a potential feedback mechanism. These results support the rationale for targeting Rac2 in the treatment of atopic dermatitis and validate the approach of Compound II in regulating Rac2 activity.

[0592] Example 12: Analysis of cell type distribution in human and mouse skin samples

[0593] To characterize the cell targets of Compound II, cell type distribution analysis was performed on Th2 human explants and MC903 mouse skin samples. Figures 17a and 17b present the results of this analysis, illustrating the distribution of various cell types affected by Compound II treatment.

[0594] The analysis revealed that Compound II affects various cell types known to express Rac2, including dendritic cells, innate lymphoid cells, T cells, and keratinocytes. This broad cellular influence supports the mechanism of action of Compound II in regulating Rac2 activity across various immune and tissue-resident cells involved in the pathogenesis of atopic dermatitis.

[0595] Example 13: Immunohistochemical analysis of Rac2 expression in atopic dermatitis

[0596] To visualize changes in Rac2 expression in atopic dermatitis, immunohistochemical staining was performed on skin tissue sections under different experimental conditions. Figure 18 presents microscopic views of these stained sections comparing control and Rac2-stained samples across different treatment conditions.

[0597] The analysis demonstrated increased Rac2 staining intensity in AD-stimulated tissues compared to control conditions, particularly in the epidermal and upper dermal regions. These results further support the upregulation of Rac2 in atopic dermatitis and validate Compound II as a relevant therapeutic target.

[0598] Example 14: Effect of Compound II on TSLP and Krt6a Levels in Atopic Dermatitis

[0599] To evaluate the effect of Compound II on key inflammatory mediators in atopic dermatitis, tissue levels of TSLP and Krt6a were measured across different treatment groups. Figures 20a and 20b present box-and-whisker plots illustrating the results of this analysis.

[0600] Treatment with Compound II resulted in a dose-dependent reduction in both TSLP and Krt6a levels. At the maximum dose of 30 mg / kg BID, Compound II reduced TSLP expression by more than 50% compared to the vehicle control, demonstrating its potent effect on this key inflammatory mediator. These results support the mechanism of action of Compound II in regulating Rac2 activity to reduce TSLP expression, a key factor in the pathogenesis of atopic dermatitis.

[0601] Example 15: Effect of Compound II on Serum IgE and Tissue Cytokine Levels

[0602] To further characterize the anti-inflammatory effects of Compound II, its effects on serum IgE, tissue IL-31, and tissue IL-13 levels were evaluated. Figures 19a, 19b, and 19c present bar graphs illustrating the effects of different treatments on these inflammatory markers.

[0603] Compound II demonstrated a dose-dependent reduction in serum IgE, tissue IL-31, and tissue IL-13 levels. At higher doses (10 and 30 mg / kg BID), Compound II showed similar or superior efficacy to upadacitinib in reducing these inflammatory markers. The ability of Compound II to regulate various inflammatory mediators supports its potential as a comprehensive therapeutic approach for atopic dermatitis through Rac2 modulation.

[0604] Example 16: Comprehensive Analysis of the Effects of Compound II on Various Cell Types and Tissues

[0605] To provide a holistic view of the effects of Compound II, a comprehensive analysis of its impact on different cell types and tissues was performed. Figures 21a, 21b, 21c, and 21d present system diagrams illustrating cell expression and functional characteristics across different cell types, accompanied by tissue-specific expression data.

[0606] Analysis revealed that Compound II affects several cell types known to express Rac2, including neutrophils, T cells, B cells, macrophages, mast cells, and Langerhans cells. By observing the effects of the compound across blood, skin, and lung tissues, its broad impact on inflammatory processes was demonstrated. These results support the mechanism of action of Compound II in regulating Rac2 activity across various cell types and tissues associated with inflammatory conditions such as atopic dermatitis and asthma.

Claims

Claim 1 A method for treating an inflammatory pathological condition comprising administering a therapeutically effective amount of a small molecule compound that regulates Rac2 activity by interfering with the binding between Rac2 and RhoGDI to a subject requiring treatment, wherein the small molecule compound reduces the expression of thymic stromal lymphocyte-producing factor (TSLP). Claim 2 A method according to claim 1, wherein the inflammatory condition is atopic dermatitis. Claim 3 A method according to paragraph 2, wherein administering the small molecule compound results in a reduction in the dermatitis severity score. Claim 4 A method according to claim 1, wherein the inflammatory condition is asthma. Claim 5 A method according to claim 1, wherein the small molecule compound reduces TSLP expression by at least 50% compared to a placebo. Claim 6 A method according to claim 1, wherein the small molecule compound is bonded to at least one of residues E171 and L155 of Rac2. Claim 7 A method according to claim 1, wherein the small molecule compound has an EC50 for binding to Rac2 of less than 10 μM. Claim 8 A method according to claim 1, wherein the small molecule compound exhibits at least three times greater binding affinity for Rac2 compared to Rac1 and Rac3. Claim 9 A method for reducing thymic stromal lymphocyte-stimulating factor (TSLP) expression in a subject, comprising administering a therapeutically effective amount of a small molecule compound to the subject that exhibits increased binding affinity for Rac2 compared to Rac1 and Rac3, wherein the small molecule compound interferes with the interaction between Rac2 and RhoGDI. Claim 10 A method according to claim 9, wherein the small molecule compound exhibits at least three times greater binding affinity for Rac2 compared to Rac1 and Rac3. Claim 11 A method according to claim 9, wherein administering the small molecule compound results in at least a 50% reduction in TSLP production compared to a placebo. Claim 12 In claim 9, the method wherein the small molecule compound has an EC50 for binding to Rac2 of less than 10 μM. Claim 13 A method according to claim 9, wherein the small molecule compound is bonded to at least one of residues E171 and L155 of Rac2. Claim 14 A method for regulating Rac2 activity, comprising administering a therapeutically effective amount of a small molecule compound that binds to Rac2 to a subject. Claim 15 A method according to claim 14, wherein the small molecule compound interferes with the interaction between Rac2 and RhoGDI. Claim 16 A method according to claim 14, wherein the small molecule compound is bonded to at least one of residues E171 and L155 of Rac2. Claim 17 In claim 14, the method wherein the small molecule compound has an EC50 for binding to Rac2 of less than 10 μM. Claim 18 A method according to claim 14, wherein administering the small molecule compound results in at least a 50% reduction in thymic stromal lymphocyte-producing factor (TSLP) expression compared to a placebo. Claim 19 A method according to claim 14, wherein the small molecule compound exhibits increased binding affinity to Rac2 compared to Rac1 and Rac3. Claim 20 A method according to claim 19, wherein the small molecule compound exhibits at least three times greater binding affinity for Rac2 compared to Rac1 and Rac3.

Images