Hydrazone compounds, pharmaceutical compositions thereof, and uses thereof

Abstract

A hydrazone compound, a pharmaceutical composition thereof and application thereof. Specifically, provided are compounds as shown in formula (III) or pharmaceutically acceptable salts thereof. The compounds have one or more of the following advantages: good water solubility; lower cytotoxicity; good inhibitory activity on Arf1 activation; strong anti-tumor immunity and induction thereof.

Description

Hydrazone compounds, pharmaceutical compositions and applications thereof

[0001] This application claims the benefit of Chinese Patent Application No. 2023118526190, filed December 28, 2023. This application incorporates the entirety of the aforementioned Chinese Patent Application. Technical Field

[0002] The present disclosure relates to hydrazone compounds, pharmaceutical compositions and applications thereof. Background Art

[0003] Cancer is the leading cause of death worldwide, killing nearly 10 million people in 2021. Despite advances in treating some types of cancer through treatments like surgery, radiation, and chemotherapy, many types are essentially incurable. Even when effective treatments are available for a particular cancer, the side effects of that treatment can be severe, significantly impairing quality of life.

[0004] Immunotherapy using immune checkpoint blockade (ICBs) and adoptive T cell therapy (ACT) has achieved landmark clinical efficacy in several advanced cancers, including melanoma, renal cell carcinoma, and lung cancer. ICBs promote PD-1 + TCF1 + CD8 + Tumor-infiltrating stem-like T cells proliferate (Im et al., 2016; Siddiqui et al., 2019) and act on different immune cell types within the tumor microenvironment (TME) to promote CD8 + T cells have an anti-tumor effect (Kurtulus et al., 2019). However, many patients do not benefit from these interventions, especially those with tumors with poor immunogenicity, such as liver cancer and triple-negative breast cancer (TNBC) (Schmid et al., 2020; Yu et al., 2021; Zhao et al., 2020). This is due to a series of potential factors, including CD8 +Systemic T cell loss, suppressed T cell activation, and poor T cell persistence or exhaustion (Bruni et al., 2020; Chen and Mellman, 2017; et al., 2019; Sharma et al., 2017; Spranger et al., 2018; Thorsson et al., 2018; Tumeh et al., 2014). These immunosuppressive factors are primarily influenced by the TME (Gupta et al., 2022). Tumor communities are composed of a heterogeneous mix of tumor cells, stromal cells (such as adipocytes and fibroblasts), immune cells, and other cells. A major effort is currently underway to improve immunotherapy by modifying the TME (Binnewies et al., 2018). Cancer cells are major players in regulating the TME (Wellenstein and de Visser, 2018). Tumors exhibit tremendous intratumoral heterogeneity. Pioneering research by John Dick and colleagues revealed that leukemia, as well as many other solid cancers, retains a hierarchical structure, with a small number of leukemia stem cells (LSCs) or cancer stem cells (CSCs), also known as tumor-initiating cells, at the apex, exhibiting unlimited self-renewal capacity, which allows tumors to regenerate and relapse despite seemingly successful treatment (Trumpp and Haas, 2022). CSCs may be the source of cells driving tumor relapse because they have the ability to enter a reversible quiescent / dormant state, resist standard chemotherapy, and form a cellular reservoir. The TME is considered the niche for CSCs and regulates their phenotypic plasticity. The TME forms an interactive and immunosuppressive environment for tumor cells and immune cells such as tumor-associated macrophages (TAMs) (Hass et al., 2020). A stem cell-like subpopulation within the tumor hemisphere of solid tumors has been found to maintain a tumor niche that allows for immunosuppression and therapy evasion (Jain et al., 2021). In TNBC, quiescent cancer stem cells (QCSCs) were recently found to constitute an immunosuppressive niche by orchestrating a local hypoxic immunosuppressive environment with dysfunctional dendritic cells (DCs), suppressive fibroblasts, reduced T cell infiltration, and enhanced T cell exhaustion (Baldominos et al., 2022). +For T cells, tumor-specific antigen peptides must be presented to the cell surface via human leukocyte antigen (HLA) class I molecules. Studies have found that several types of CSCs downregulate the expression of HLA class I molecules (Muller et al., 2020). CSCs also express CD47α, which inhibits macrophage phagocytosis by interacting with signal-regulating proteins (Muller et al., 2020). Therefore, developing new strategies to eradicate CSCs and alter their immunosuppressive environment will be key to improving immunotherapy responses in cancer treatment.

[0005] For many years, the inventors have been dedicated to the study of stem cells and cancer stem cells. Several years ago, the inventors performed genome-wide RNAi screening in Drosophila intestinal stem cells (ISCs) and discovered a set of genes (including genes in the Arf1-mediated lipid metabolism pathway) whose knockdown induces cytokine release, affects their microenvironment, and ultimately leads to stem cell death. The specific mechanisms of action of these genes vary, but a common feature is that their knockdown first releases factors from stem and cancer stem cells, alters the microenvironment, activates neighboring cells or immune cells, and then feeds back to kill the stem cells or CSCs (Aggarwal et al., 2022; Singh et al., 2016; Wang et al., 2020; Zeng et al., 2015). Deletion of these genes in CSCs releases cytokines, transforming the TME from an immunosuppressive to an immunostimulatory microenvironment (Wang et al., 2020).

[0006] ADP-ribosylation factor 1 (Arf1), a member of the human Arf gene family, encodes a small guanine nucleotide-binding protein that plays an important role in vesicle trafficking and lipid metabolism (D'Souza-Schorey and Chavrier, 2006; Donaldson and Jackson, 2011; Kazmalek et al., 2017; Wilfling et al., 2014). Previous studies have shown that Arf1 is highly expressed in a variety of human cancers, including breast, hepatocellular, colorectal, and prostate cancers (Casalou et al., 2016; Wang et al., 2020). Upregulation of Arf1 is negatively correlated with prognosis in cancer patients (Lang et al., 2017; Wang et al., 2020).

[0007] Knocking out the Arf1 pathway in CSCs triggers a metabolic stress chain reaction, starting with mitochondrial damage and endoplasmic reticulum (ER) stress, and then exposing damage-associated molecular patterns (DAMPs: CALR, HMGB1, ATP), thereby changing the tumor microenvironment and activating a systemic antitumor immune response, thereby destroying the tumor (Wang et al., 2020). The inventors recently tested two new Arf1 inhibitors (DU101, DU102; described in our previous patents: WO2022 / 028429A1, PCT / CN2021 / 110373) and revealed their novel antitumor mechanisms. The inventors demonstrated that the two newly designed Arf1 inhibitors have low toxicity and potent antitumor activity in mouse and patient-derived liver and melanoma tumor xenograft (PDX) models. Treatment with the inhibitors significantly increased CD8 + Infiltration and activation of T cells and induction of tumor-associated stem cell-like TCF1 through the S1P-S1PR pathway + CD4 + CD8 + Formation of double-positive T (DPT) cells.

[0008] Two recent studies have found that tumor-associated CD4 / CD8 double-positive T (DPT) cells have potent antitumor activity in hepatocellular carcinoma (HCC) and other tumors (Schad et al., 2022; Zheng et al., 2020). Since early DPTs are mainly present in the thymus, tumor-associated DPT cells may be derived from infiltrating CD4 + or CD8 + Single-positive T cells express both exhaustion and activation markers. DPT cells demonstrate a robust immune response and are in an active anti-tumor state. Furthermore, HCC patients enriched in DPT cells are positively correlated with better treatment outcomes, suggesting that DPT cells are superior anti-tumor T cells.

[0009] If the tumor persists, active CD8 + T cells are typically exhausted, so maintaining a functional tumor-targeting CD8 + The T cell pool is crucial. In cancer patients receiving immunotherapy, the duration of functional anti-tumor T cells is directly correlated with better treatment outcomes (Robbins et al., 2004). Studies have shown that ICBs inhibit tumors not by reversing T cell exhaustion, but by promoting the proliferation of stem-like T cell subsets. Stem-like T cells then generate fresh, effective T cells to replenish the exhausted T cells in the tumor. Stem-like CD8 +T cells maintain superior responses to ICBs and ACT in animal models of cancer and in patients with cancer (Baharon et al., 2021; Eberhardt et al., 2021; Galletti et al., 2020; Hashimoto et al., 2022; Im et al., 2016; Jansen et al., 2019; Kurtulus et al., 2019; Krishna et al., 2020; Mo et al., 2021; Prokhnevska et al., 2023; Siddiqui et al., 2019; Wu et al., 2020; Yost et al., 2019).

[0010] Our data suggest that treatment with an Arf1 inhibitor enriches stem cell-like CD4 + CD8 + DPT cell population, which combines stem cell-like T cells and CD4 + CD8 + The superior properties of these two T cell subsets of DPT cells may have more powerful anti-tumor activity. Exploring the properties of this special T cell population may further improve the efficiency of T cell immunotherapy. Therefore, treatment with a single agent can overcome the three obstacles of cancer immunotherapy: increasing CD8 + Arf1 infiltration promotes T cell activation and blocks T cell dysfunction or exhaustion. Treatment with these inhibitors shifts the tumor immune microenvironment from cold to warm by reprogramming lipid metabolism. These inhibitors may provide new cancer immunotherapy strategies and significantly expand the proportion of cancer patients suitable for immunotherapy. New Arf1 inhibitors can not only be used to treat tumors with poor immunogenicity, but also develop more effective T cell-based immunotherapies by combining the inhibitors with ICBs or ACT.

[0011] However, the poor water solubility and permeability of DU101 and DU102 greatly limit their transformative potential. Therefore, there is an urgent need to develop and identify new Arf1 inhibitors with good water solubility and induce strong anti-tumor immunity. Summary of the Invention

[0012] This application is based on the inventors' unexpected discovery that ablation of the COPI / Arf1-lipolysis β-oxidation pathway can eradicate cancer stem cells and induce DAMPs-mediated anti-tumor immune responses, and the development and identification of a new class of hydrazone compounds, their pharmaceutical compositions, and their uses. The hydrazone compounds of this application have good water solubility and induce strong anti-tumor immunity. Therefore, the hydrazone compounds of this application are particularly suitable for treating and / or preventing diseases related to Arf1 pathway activity; treating refractory, recurrent, or metastatic cancers; selectively killing cancer cells with specific dosing regimens; and targeting cancer stem cells (CSCs) to induce anti-tumor immune responses by inhibiting the Arf1 pathway, particularly the COPI / Arf1-lipolysis β-oxidation pathway.

[0013] In one aspect, the present application provides a compound of formula (III) or a pharmaceutically acceptable salt thereof:

[0014] in, represents a double bond, which is in Z form, E form, or a mixture of Z and E forms;

[0015] X3, X4, X5, X6 and X9 are each independently C(R x ) or N, R x independently H, D (deuterium), CN, OH, NH2, NHSO2C 1- 6 alkyl, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy or C 1-6 haloalkoxy;

[0016] X7 and X8 are each independently C(R x ) or N, R x independently H, D (deuterium), CN, OH, NH2, NHSO2C 1-6 Alkyl, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy or C 1-6 haloalkoxy;

[0017] Alternatively, X7 is C(R x ), X8 is C(R x ), R in X7 x and R in X8 x Together with the atoms to which they are attached, they form a group optionally substituted by 1, 2 or 3 R eA substituted ring D, wherein the ring D is a C5-C6 cycloalkene, a 5-6 membered heterocycloalkene, a benzene ring or a 5-6 membered heteroaromatic ring, wherein the heteroatoms in the 5-6 membered heterocycloalkene and the 5-6 membered heteroaromatic ring are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3;

[0018] Ring A is a C5-C6 cycloalkene, a 5-6 membered heterocycloalkene, a benzene ring or a 5-6 membered heteroaromatic ring, wherein the heteroatoms in the 5-6 membered heterocycloalkene and the 5-6 membered heteroaromatic ring are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3;

[0019] n is 1, 2, or 3;

[0020] R1 is independently H, D, CN, OH, halogen, biphenyl, -C(O)-OC 1-6 Alkyl, optionally substituted by 1, 2 or 3 R a Substituted C 1-6 Alkyl, optionally substituted by 1, 2 or 3 R b Substituted C 1-6 Alkoxy, optionally substituted by 1, 2 or 3 R a Substituted C 3-10 Cycloalkyl, optionally substituted by 1, 2 or 3 R a substituted 3-10 membered heterocycloalkyl, optionally substituted by 1, 2 or 3 R c Substituted C 6-14 Aryl or optionally substituted by 1, 2 or 3 R c A substituted 5-14 membered heteroaryl group, wherein the heteroatoms in the 5-14 membered heteroaryl group are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3;

[0021] Or R1 on two adjacent carbon atoms and the carbon atoms to which they are connected together form a group optionally substituted by 1, 2 or 3 R d Substituted C 5-8 Cycloalkene or optionally 1, 2 or 3 R d Substituted 5- to 8-membered heterocyclic olefins, wherein the heteroatoms in the 5- to 8-membered heterocyclic olefins are selected from one or more of N, O, and S, and the number of heteroatoms is 1, 2, or 3;

[0022] R a 、R b 、R c 、R d and R e are independently D, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C1- 6-haloalkoxy, -C(O)-C 1-6 Alkyl, -C(O)-OC 1-6 Alkyl, -C(O)-C 3-6 Cycloalkyl, oxo (=O), -NH2, optionally substituted by 1, 2 or 3 R a-1 Substituted-C(O)-C 6-14 Aryl or any 1, 2 or 3 R a-1 substituted-C(O)-5-14 membered heteroaryl;

[0023] R a-1 are each independently halogen;

[0024] The compound of formula (I) is not

[0025] In some embodiments, each of the C 1-6 Alkyl and substituted C 1-6 C in alkyl 1-6 Alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl or isopentyl; for example, methyl, ethyl, tert-butyl, isopropyl, n-propyl, n-butyl or n-pentyl; preferably methyl.

[0026] In some embodiments, each of the C 1-6 Alkoxy and substituted C 1-6 C in alkoxy 1-6 Alkoxy is independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy; for example, methoxy or ethoxy.

[0027] In some embodiments, the 5-6 membered heteroaromatic rings in each of the 5-6 membered heteroaromatic rings and the substituted 5-6 membered heteroaromatic rings are independently 5 membered heteroaromatic rings having 1 or 2 heteroatoms; for example,

[0028] In some embodiments, each of the 5-6 membered heterocyclic olefins is a 6 membered heterocyclic olefin wherein the heteroatom is N and the number of heteroatoms is 1; for example,

[0029] In some embodiments, each of the C 3-10 Cycloalkyl and substituted C 3-10 C in cycloalkyl 3-10 Cycloalkyl is independently cyclohexyl or

[0030] In some embodiments, the 3-10 membered heterocycloalkyl and substituted 3-10 membered heterocycloalkyl groups are independently 3-6 membered heterocycloalkyl groups; preferably, the heteroatom is O and the number of heteroatoms is 1 6 membered heterocycloalkyl group; for example,

[0031] In some embodiments, each of the C 6-14 Aryl and substituted C 6-14 C in aromatic group 6-14 Aryl is independently C 6-10 Aryl; for example,

[0032] In some embodiments, the 5-14 membered heteroaryl and the substituted 5-14 membered heteroaryl are each independently a 5-10 membered heteroaryl; preferably a 5-10 membered heteroaryl having 1 heteroatom; for example,

[0033] In some embodiments, each of the C 5-8 Cycloolefins

[0034] In some embodiments, the 5-8 membered heterocyclic olefins and substituted 5-8 membered heterocyclic olefins are independently 5-6 membered heterocyclic olefins having a heteroatom of N and 1 heteroatom; for example,

[0035] In some embodiments, each said halogen is independently fluoro, chloro, bromo, or iodo; for example, fluoro.

[0036] In some embodiments, each said halo is independently fluoro, chloro, bromo, or iodo.

[0037] In some embodiments, each of the C 2-6 Alkenyl and substituted C 2-6 C in alkenyl 2-6 Alkenyl is independently C 2-4 alkenyl; for example,

[0038] In some embodiments, X3 is N.

[0039] In some embodiments, X4 is N.

[0040] In some embodiments, R x is independently H, OH or NHSO2Me; preferably H.

[0041] In some embodiments, X5 is C(R x ); for example, CH.

[0042] In some embodiments, X6 is C(R x ); for example, CH.

[0043] In some embodiments, X7 is C(R x );For example,

[0044] In some embodiments, X8 is C(R x ); for example, CH.

[0045] In some embodiments, X7 is C(R x ), X8 is C(R x ), R in X7 x and R in X8 x Together with the atoms to which they are attached, they form a ring D, wherein the ring D is a 5-6 membered heteroaromatic ring; for example, Among them, 1 is connected to X6, and 2 is connected to X9.

[0046] In some embodiments, X9 is C(R x ); for example, CH.

[0047] In some embodiments, Ring A is a 5-6 membered heterocycloalkene or a 5-6 membered heteroaryl ring.

[0048] In some embodiments, n is 1 or 2.

[0049] In some embodiments, R c are independently halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 1-6 Haloalkyl or C 1-6 Alkoxy; for example, methyl, methoxy, ethyl, trifluoromethyl, tert-butyl, fluoro, isopropyl, n-propyl or

[0050] In some embodiments, R d Independently C 1-6 Alkyl, -C(O)-5-14 membered heteroaryl, -C(O)-C 1-6 Alkyl, -C(O)-OC 1-6 Alkyl, -C(O)-C 3-6 Cycloalkyl or optionally substituted by 1, 2 or 3 R a-1 Substituted-C(O)-C 6-14 Aryl; for example, methyl, ethyl, n-propyl,

[0051] In some embodiments, each R1 is independently H, biphenyl, -C(O)-OC 1-6 Alkyl, C1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocycloalkyl, optionally substituted by 1, 2 or 3 R c Substituted C 6-14 Aryl or optionally substituted by 1, 2 or 3 R c substituted 5-14 membered heteroaryl; for example, H, methyl, Propyl, butyl, pentyl, Ethyl,

[0052] In some embodiments, R1 on two adjacent carbon atoms together with the carbon atom to which they are attached form a group optionally substituted by 1, 2 or 3 R d Substituted C 5-8 Cycloalkene or optionally 1, 2 or 3 R d substituted 5-8 membered heterocyclic olefins; for example, optionally substituted with 1, 2 or 3 R d Replaced For example,

[0053] Preferably, 1, 2 or 3 R d substituted 5-8 membered heterocyclic olefins; for example, optionally substituted with 1, 2 or 3 R d Replaced Further example,

[0054] In some embodiments, for in for The definition of R1 is as described in any embodiment of the present invention, for When R1 on two adjacent carbon atoms does not form a ring with the atoms to which they are attached; for example,

[0055] In some embodiments, for in for Wherein, R1 is defined as described in any embodiment of the present invention, and R1 on two adjacent carbon atoms forms a ring with the atoms to which they are connected; for example,

[0056] In some embodiments, for For example,

[0057] In another aspect, the present application provides a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0058] in,

[0059] X1, X2 are selected from C, N, O or S;

[0060] represents a single bond or a double bond;

[0061] represents a double bond, which can be of Z or E form;

[0062] X3, X4, X5, X6, X7, X8 and X9 are each independently selected from C(R x ) or N, R x independently selected from H, D (deuterium), CN, OH, NH2, NHSO2Me, halogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy and C 1-6 a group consisting of a haloalkoxy group;

[0063] R1, R2, R3 are each independently absent or selected from H, D, CN, OH, halogen, biphenyl, -C(O)-OC 1-6 Alkyl, optionally substituted with 1-3 R a C 1-6 Alkyl, optionally substituted with 1-3 R b C 1-6 Alkoxy, optionally substituted with 1-3 R a C 3- 6-cycloalkyl, optionally substituted with 1-3 R a C 3-6 Heterocycloalkyl, optionally substituted with 1-3 R c C 6-14 Aryl, optionally substituted with 1-3 R c C 6-14 or R1 and R2 together with the carbon atom to which they are attached form a group optionally substituted with 1-3 R d C 5-8 a carbocyclic ring or a 5- to 8-membered heterocyclic ring, the 5- to 8-membered heterocyclic ring optionally containing one or more heteroatoms selected from N, O and S;

[0064] Ring D is absent or optionally substituted with 1-3 Re C 5-6 Carbocyclic or 5- to 6-membered heterocyclic ring;

[0065] Each time it appears, each R a 、R b 、R c 、R d 、R e Each independently selected from D, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, -C(O)-C 1-6 Alkyl, -C(O)-OC 1-6 Alkyl, -C(O)-C 1-6 Cycloalkyl, -C(O)-C 6-14 The group consisting of aryl, oxo (=O) and -NH2.

[0066] When used in this article, Represents a single bond or a double bond. Those skilled in the art will understand that when “ When the "substitution" is a single bond or a double bond, the resulting compound molecule should satisfy the valence bond theory.

[0067] In some embodiments, Ring A is selected from the following ring structures:

[0068] In some embodiments, R1, R2, and R3 are each independently absent or selected from H, D, CN, OH, halogen, biphenyl, -C(O)-OC 1-6 Alkyl, optionally substituted with 1-3 R a C 1-6 Alkyl, optionally substituted with 1-3 R b C 1-6 Alkoxy, optionally substituted with 1-3 R a C 3-10 Cycloalkyl, optionally substituted with 1-3 R a C 3-6 Heterocycloalkyl, optionally substituted with 1-3 R c C 6-14 Aryl and optionally substituted with 1-3 R c C 6-14The group consisting of heteroaryl, for example, H, D, CN, OH, F, Cl, Br, I, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, adamantyl, cyclohexyl, tetrahydrofuranyl, methoxy, ethoxy, propoxy, butoxy, methoxycarbonyl, ethoxycarbonyl, phenyl, tolyl, ethylphenyl, methoxyphenyl, trifluoromethylphenyl, fluorophenyl, dimethylphenyl, isopropylphenyl, isobutenylphenyl, epoxybutylphenyl, cyclohexylphenyl, biphenyl or naphthyl.

[0069] In some embodiments, Ring A is selected from the following ring structures:

[0070] In some embodiments, R1 and R2 together with the carbon atom to which they are attached form a C 5-8 A carbocyclic ring or a 5- to 8-membered heterocyclic ring, for example, selected from the following ring structures:

[0071] Preferred

[0072] in, represents a single bond or a double bond;

[0073] The C 5-8 Carbocyclic or 5- to 8-membered heterocyclic rings are optionally substituted with 1-3 R d .

[0074] In some embodiments, R1 and R2 together with the carbon atom to which they are attached form a C 5-8 When it is a carbocyclic ring or a 5- to 8-membered heterocyclic ring, the fused ring structure formed together with Ring A is selected from the following structures:

[0075] It is optionally substituted with 1-3 R d .

[0076] In some embodiments, when R1 and R2 together with the carbon atom to which they are attached form C 5-8 When it is a carbocyclic ring or a 5- to 8-membered heterocyclic ring, the fused ring structure formed together with Ring A is selected from the following structures:

[0077] In some embodiments, Ring B is selected from the following structures:

[0078] For example,

[0079] In some embodiments, X5, X6, X7, X8 and X9 are each independently selected from C(R x ).

[0080] In some embodiments, when Ring D is absent, Ring C is optionally substituted with 1-5 R x ; Ring C, for example, 1, 2, 3, 4 or 5 R x Substituted benzene ring.

[0081] In some embodiments, when Ring D is present, Ring C is optionally substituted with 1-3 R x ; Ring C, for example, 1, 2 or 3 R x Substituted benzene ring.

[0082] In some embodiments, any one of X5, X6, X7, X8 and X9 is N, and the others are C(R x ).

[0083] In some embodiments, when Ring D is absent, Ring C is optionally substituted with 1-4 R x ; Ring C, for example, 1, 2, 3 or 4 R x Substituted pyridine ring.

[0084] In some embodiments, when Ring D is present, Ring C is optionally substituted with 1-2 R x ; Ring C, for example, 1 or 2 R x Substituted pyridine ring.

[0085] In some embodiments, Ring C is selected from the following ring structures:

[0086] For example,

[0087] In some embodiments, Ring D is optionally substituted with 1-3 R e , such as 1, 2 or 3 R e Substituted ring structures:

[0088] Where "1" indicates the connection direction with X7 of ring C, and "2" indicates the connection direction with X8 of ring C; for example,

[0089] In some specific embodiments, R e C 1-4 Alkyl, C 1-4 Halogenated alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy or oxo (=O), such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, trifluoromethyl, trifluoroethyl (such as -CH2CF3), methoxy, ethoxy, propoxy, butoxy, C 1-4 Haloalkoxy or oxo (=O).

[0090] In some embodiments, Ring D is selected from the following ring structures:

[0091] For example,

[0092] In some embodiments, Rings C and D collectively form the following fused ring structure:

[0093] Ring C is optionally substituted with 1-3 R x , Ring D is optionally substituted with 1-3 R e .

[0094] In some embodiments, Rings C and D collectively form the following fused ring structure:

[0095] In some embodiments, the present application provides a compound of formula (II) or a pharmaceutically acceptable salt thereof:

[0096] wherein R1, R2, X5, X6 and X9 are as defined above.

[0097] In some embodiments, the compound of formula (I) or formula (III) is selected from the following compounds:

[0098] in, represents a double bond, representing Z-form, E-form or a mixture of Z-form and E-form; preferably, is of formula E; the compound represented by formula (I) or formula (III) is any of the following compounds:

[0099] In one aspect, the present application provides a pharmaceutical composition comprising a compound of formula (I) or formula (III) described herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0100] In one aspect, the present application provides a method for inhibiting intracellular Arf1 pathway activity in a cell, comprising administering an effective amount of a compound of formula (I) or formula (III), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above to the cell, such that the Arf1 pathway activity in the cell is reduced.

[0101] In some embodiments, the cell is a progenitor cell, a stem cell, a cancer stem cell, or a cancer cell.

[0102] In some embodiments, the methods induce cell death at the cellular level.

[0103] In some embodiments, the method is performed in vitro.

[0104] In some embodiments, the method is performed in vivo.

[0105] In one aspect, the present application provides use of a compound of formula (I) or formula (III), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above in the preparation of an agent for inhibiting intracellular Arf1 pathway activity in a cell.

[0106] In some embodiments, the cell is a progenitor cell, a stem cell, a cancer stem cell, or a cancer cell.

[0107] In some embodiments, the agent induces cell death at the cellular level.

[0108] In some embodiments, the agent is administered to the cell in vitro.

[0109] In some embodiments, the method is administered to the cells in vivo.

[0110] In one aspect, the present application provides a compound of formula (I) or formula (III), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above for use in treating or preventing a disease associated with Arf1 pathway activity in a subject.

[0111] In some embodiments, the disease associated with Arf1 pathway activity is cancer or a tumor.

[0112] In some embodiments, the cancer or tumor is selected from the group consisting of breast cancer, head and neck cancer, lung cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, renal cell carcinoma, melanoma, hepatocellular carcinoma, cervical cancer, sarcoma, brain tumor, gastric cancer, multiple myeloma, leukemia, and lymphoma.

[0113] In some embodiments, the disease associated with Arf1 pathway activity is selected from the group consisting of autoimmune diseases, inflammatory diseases, inflammatory bowel disease, arthritis, autoimmune demyelinating disorders, Alzheimer's disease, amyotrophic lateral sclerosis, stroke, ischemia-reperfusion injury, multiple sclerosis and other neurodegenerative diseases.

[0114] In some embodiments, the subject is a mammal, such as a human.

[0115] In some embodiments, the compound of formula (I) or formula (III), their pharmaceutically acceptable salts, or the pharmaceutical composition described above are administered in combination with at least one immune checkpoint inhibitor, such as an anti-PD-1 antibody.

[0116] In some embodiments, the compound of formula (I) or formula (III) described herein, their pharmaceutically acceptable salts, or the pharmaceutical composition as described above are administered simultaneously with at least one immune checkpoint inhibitor, such as an anti-PD-1 antibody, or administered sequentially in any order.

[0117] In one aspect, the present application provides the use of the compound of formula (I) or formula (III), their pharmaceutically acceptable salts, or the pharmaceutical composition as described above in the preparation of a medicament for treating or preventing a disease associated with Arf1 pathway activity in a subject.

[0118] In some embodiments, the disease associated with Arf1 pathway activity is cancer or a tumor.

[0119] In some embodiments, the cancer or tumor is selected from the group consisting of breast cancer, head and neck cancer, lung cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, renal cell carcinoma, melanoma, hepatocellular carcinoma, cervical cancer, sarcoma, brain tumor, gastric cancer, multiple myeloma, and leukemia and lymphoma.

[0120] In some embodiments, the disease associated with Arf1 pathway activity is selected from the group consisting of autoimmune diseases, inflammatory diseases, inflammatory bowel disease, arthritis, autoimmune demyelinating disorders, Alzheimer's disease, amyotrophic lateral sclerosis, stroke, ischemia-reperfusion injury, multiple sclerosis and other neurodegenerative diseases.

[0121] In some embodiments, the subject is a mammal, such as a human.

[0122] In one aspect, the present application provides a kit for treating or preventing a disease associated with Arf1 pathway activity in a subject, the kit comprising a compound of formula (I) or formula (III) as described herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above; a container; and optionally a package insert or label indicating treatment or prevention.

[0123] In some embodiments, the disease associated with Arf1 pathway activity is cancer or a tumor.

[0124] In some embodiments, the cancer or tumor is selected from the group consisting of breast cancer, head and neck cancer, lung cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, renal cell carcinoma, melanoma, hepatocellular carcinoma, cervical cancer, sarcoma, brain tumor, gastric cancer, multiple myeloma, leukemia, and lymphoma.

[0125] In some embodiments, the disease associated with Arf1 pathway activity is selected from the group consisting of autoimmune diseases, inflammatory diseases, inflammatory bowel disease, arthritis, autoimmune demyelinating disorders, Alzheimer's disease, amyotrophic lateral sclerosis, stroke, ischemia-reperfusion injury, multiple sclerosis and other neurodegenerative diseases.

[0126] In some embodiments, the subject is a mammal, such as a human.

[0127] In some embodiments, the kit further comprises at least one immune checkpoint inhibitor, such as an anti-PD-1 antibody.

[0128] In one aspect, the present application provides a kit for diagnosing a disease associated with Arf1 pathway activity in a subject, comprising at least one compound of formula (I) or formula (III) described herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above as a test reagent.

[0129] In some embodiments, the kit detects at least one biomarker indicative of the presence of a disease associated with Arf1 pathway activity.

[0130] In some embodiments, the kit can detect Arf1 GTPase activity, serve as a reporter gene for detecting lipolytic activity, detect lipid droplet formation, detect autophagic activity, or serve as an upstream or downstream surrogate regulator of Arf1 activity or function.

[0131] In some embodiments, the disease associated with Arf1 pathway activity is cancer or a tumor.

[0132] In some embodiments, the cancer or tumor is selected from the group consisting of breast cancer, head and neck cancer, lung cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, renal cell carcinoma, melanoma, hepatocellular carcinoma, cervical cancer, sarcoma, brain tumor, gastric cancer, multiple myeloma, leukemia, and lymphoma.

[0133] In some embodiments, the disease associated with Arf1 pathway activity is selected from the group consisting of autoimmune diseases, inflammatory diseases, inflammatory bowel disease, arthritis, autoimmune demyelinating disorders, Alzheimer's disease, amyotrophic lateral sclerosis, stroke, ischemia-reperfusion injury, multiple sclerosis and other neurodegenerative diseases.

[0134] In some embodiments, the subject is a mammal, such as a human.

[0135] In one aspect, the present application provides the use of the compound of formula (I) or formula (III), their pharmaceutically acceptable salts, or the pharmaceutical composition as described above in the preparation of a therapeutic vaccine for blocking tumor development.

[0136] In some embodiments, the tumor is selected from the group consisting of breast cancer, head and neck cancer, lung cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, renal cell carcinoma, melanoma, hepatocellular carcinoma, cervical cancer, sarcoma, brain tumor, gastric cancer, multiple myeloma, leukemia, and lymphoma.

[0137] In one aspect, the present application provides a method for preparing a therapeutic vaccine for blocking tumor development, comprising contacting a compound of formula (I) or formula (III) described herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above with tumor stem cells, thereby converting the tumor stem cells into a therapeutic vaccine.

[0138] In one aspect, the present application provides a vaccination method, comprising administering the therapeutic vaccine as described above to a subject.

[0139] In some embodiments, the subject is a mammal, such as a human.

[0140] In one aspect, the present application provides a method for killing cells and inducing an anti-tumor immune response, the method comprising inhibiting the activity of at least one Arf1 in the cell, particularly the activity of the COPI / Arf1-lipolysis β-oxidation pathway, by an Arf1 pathway inhibitor.

[0141] In some embodiments, the cell is a progenitor cell, a stem cell, a cancer stem cell, or a cancer cell.

[0142] In some embodiments, the Arf1 pathway inhibitor is selected from the group consisting of a small molecule Arf1 inhibitor, an RNAi agent directed against Arf1, an antisense agent directed against Arf1, a peptidomimetic Arf1 inhibitor, and a DNA binding deoxynucleotide inhibitor containing the target protein Arf1.

[0143] In some embodiments, the small molecule Arf1 inhibitor is a compound of Formula (I) or Formula (III) as described herein, or a pharmaceutically acceptable salt thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0144] Figures 1-3 show the inhibition of Arf1 activation by the compounds of the present application

[0145] Figures 4-8 show the inhibitory effects of the compounds of the present application on tumor growth in Drosophila.

[0146] Figures 9-10 show the inhibitory effects of the compounds of the present application on CT26 colon cancer in BALB / c mice. DETAILED DESCRIPTION

[0147] Certain embodiments will now be described in detail, and examples are illustrated in the accompanying specific embodiments. Although only the embodiments of enumeration are described, it should be understood that they are not intended to limit the present invention to these embodiments. On the contrary, the present invention is intended to encompass all alternatives, modifications, and equivalents, which may be included within the scope of this application. Those skilled in the art will recognize that many methods and materials similar or equivalent to those described herein can be used for practice of the present invention. The present invention is in no way limited to described methods and materials. If one or more incorporated documents and similar materials differ from or contradict the present disclosure, including but not limited to defined terms, term usage, described technology, etc., the present disclosure shall prevail.

[0148] It will be appreciated that certain features of the present invention, which, for clarity, are described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the present invention, which, for brevity, are described in the context of a single embodiment, may also be provided separately or in any suitable subcombination.

[0149] definition

[0150] Terms used but not defined herein have their ordinary meanings, and the meanings of such terms are independent at each occurrence thereof. In addition, unless otherwise indicated, the following definitions apply to the meaning and scope of the various terms of the present invention.

[0151] As used herein, the terms “comprising” and “including” are intended to specify the presence of stated features, integers, components or steps, but they do not preclude the presence or addition of one or more other features, integers, components, steps or groups thereof.

[0152] The definitions of specific functional groups and chemical terms are described in more detail below. For the purposes of the present invention, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition, inside cover, and specific functional groups are generally defined as described therein. In addition, the general principles of organic chemistry as well as specific functional moieties and reactivities are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March, March's Advanced Organic Chemistry, 5th edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; Carruthers, Some Modem Methods of Organic Synthesis, 3rd edition, Cambridge University Press, Cambridge, 1987.

[0153] All ranges cited herein are inclusive unless expressly stated otherwise.

[0154] When a range of values ​​is listed, it is intended that every value and subrange within that range be included. For example, “C 1-6 "Aims to cover C1, C2, C3, C4, C5, C6, C 1-6 、C 1-5 、C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-5 、C 2-4 、C 2-3 、C 3-6 、C 3-5 、C 3-4 、C 4-6 、C 4-5 and C 5-6 .

[0155] When any variable occurs more than one time in any constituent, for example, in Formula (I), Formula (III), or any other formula depicting and describing the compounds of the present application, its definition on each occurrence is independent of its definition at every other occurrence. Furthermore, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0156] As used herein, the terms "cancer stem cell" and "CSC" are interchangeable. The CSC is of mammalian origin, and in some embodiments, the CSC is of human origin, but is not limited thereto. The definition and functional characteristics of cancer stem cells are: 1) a population of tumor cells with extensive proliferative capacity; 2) the ability to undergo asymmetric cell division to produce one or more differentiated offspring with reduced proliferation or developmental potential; 3) the ability to undergo symmetric cell division that is self-renewal or self-sustaining. Other common methods for characterizing CSCs include examining morphology, cell surface markers, transcriptional profiles, and drug responses. In the research literature, CSCs are also referred to as tumor / cancer initiating cells, cancer stem-like cells, stem-like cancer cells, highly tumorigenic cells, tumor stem cells, solid tumor stem cells, drug-surviving cells (DSCs), drug-resistant cells (DRCs), or super malignant cells.

[0157] As used herein, the terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals in which a cell population is characterized by unregulated cell growth. As used herein, the terms "cancer cell" and "tumor cell" refer to the total number of cells from a tumor, including tumorigenic stem cells (cancer stem cells) and non-tumorigenic cells that make up the majority of the tumor cell population. Examples of cancer include, but are not limited to, carcinomas, lymphomas, blastomas, sarcomas, and leukemias. More specific examples of such cancers include squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer, lung adenocarcinoma, squamous cell lung cancer, peritoneal cancer, hepatocellular carcinoma, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, liver cancer, breast cancer, colon cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer, liver cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, and various types of head and neck cancer.

[0158] As used herein, the term "tumor" refers to any mass of tissue resulting from excessive growth or proliferation of cells, whether benign (non-cancerous) or malignant (cancerous), including precancerous lesions.

[0159] As used herein, the term "metastasis" refers to the process by which cancer spreads or metastasizes from its site of origin to other areas of the body as similar cancerous lesions develop in new locations. "Metastatic" cells are cells that have lost their adhesive contacts with neighboring cells and migrated from the primary site of the disease via the blood or lymph to invade neighboring body structures.

[0160] As used herein, the term "subject" refers to any animal (e.g., mammal), including but not limited to humans, non-human primates, rodents, etc., that is to be the recipient of a particular treatment. Generally, the terms "subject" and "patient" are used interchangeably herein to refer to a human subject.

[0161] As used herein, the terms "treat" or "alleviate" refer to: 1) a cure, slowdown, relief of symptoms and / or halting of progression of a diagnosed pathological condition or disease; 2) a prophylactic or therapeutic method that prevents or slows the development of a target pathological condition or disease. Therefore, those in need of treatment include those already suffering from the disease; those susceptible to such disease; and those in need of prevention. A subject is successfully "treated" according to the methods of the present application if the patient exhibits one or more of the following: a decrease or complete absence of cancer cells; a reduction in tumor size; inhibition or absence of cancer cell infiltration into peripheral organs (these include the spread of cancer cells to soft tissue and bone); inhibition or absence of tumor metastasis; inhibition or absence of tumor growth; relief of one or more symptoms associated with a particular cancer; reduction in morbidity and mortality; and improvement in quality of life.

[0162] As used herein, the term "inhibit," when used in the context of a biological activity, refers to downregulation of a biological activity that may reduce or eliminate a target function, such as the production of a protein or the phosphorylation of a molecule. In some embodiments, inhibition may refer to a reduction in the targeted activity by about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95%. When used in the context of a disorder or disease, the term may refer to an effect achieved in preventing the onset of symptoms, alleviating symptoms, or eliminating the disease, condition, or disorder.

[0163] As used herein, the term "lipolysis" refers to the breakdown of lipids and involves the hydrolysis of triglycerides into glycerol and free fatty acids. Lipolysis occurs primarily in adipose tissue and is used to mobilize stored energy during fasting or exercise.

[0164] As used herein, the term "β-oxidation" refers to the metabolic process by which fatty acid molecules are broken down in the cytosol of prokaryotes and the mitochondria of eukaryotes to produce acetyl-CoA, which enters the citric acid cycle, and NADH2 and FADH, which serve as coenzymes in the electron transport chain. β-oxidation is so named because the β carbon of the fatty acid is oxidized to a carbonyl group. β-oxidation is primarily catalyzed by the mitochondrial trifunctional protein, a multienzyme complex associated with the inner mitochondrial membrane, although some fatty acids are also oxidized in peroxisomes.

[0165] As used herein, the term "deuterated" refers to a compound or group in which one or more hydrogen atoms are replaced by deuterium. When a compound or group is deuterated, one, two, three, or even more hydrogen atoms on the compound or group may be replaced by deuterium, until all hydrogen atoms on the compound or group are replaced by deuterium. In this case, the compound or group is referred to as a "perdeuterated compound or group."

[0166] In some embodiments, at deuterated positions, the deuterium isotope abundance is greater than the natural deuterium isotope abundance (0.015%), preferably greater than 50%, more preferably greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, greater than 99%, greater than 99.5%, or 100%.

[0167] In some cases, such as when "hydrogen" and "deuterium" appear in a statement as parallel alternatives or when "hydrogen" is replaced by "deuterium", the term "hydrogen" refers to the hydrogen isotope " 1 hydrogen( 1 H)", while the term "deuterium" refers to the hydrogen isotope " 2 hydrogen( 2 H)"; or it is understood that at that position of the compound, hydrogen in a state where its various isotopic abundances are naturally present is replaced by deuterium in a state where deuterium is present at a greater abundance than the natural deuterium isotopic abundance (e.g., a deuterium abundance of greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, greater than 99%, greater than 99.5%, or 100%).

[0168] As used herein, the term "hydrocarbyl" refers to a chemical group comprising hydrogen and carbon. A hydrocarbyl group may be substituted or unsubstituted. A hydrocarbyl group may be unsaturated, saturated, branched, unbranched, cyclic, polycyclic or heterocyclic, and includes alkyl, alkenyl and alkynyl groups. A hydrocarbyl group may be fully saturated, monounsaturated or polyunsaturated, and may include divalent and polyvalent groups, for example, when a divalent group, it may be referred to as a "hydrocarbylene group". A hydrocarbyl group has a specified number of carbon atoms (i.e., C1-C 10 "alkyl" means 1 to 10 carbon atoms, including 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 carbon atoms. In some embodiments, the hydrocarbon group contains one or more, for example, 1, 2, 3, or 4, unsaturated carbon-carbon double bonds (-C=C-), carbon-carbon triple bonds (-C≡C-), and / or any combination thereof. In some embodiments, the hydrocarbon group may be deuterated, also known as a "deuterated hydrocarbon group," i.e., one, two, three, or even more hydrogen atoms on the hydrocarbon group may be replaced by deuterium, until all hydrogen atoms on the hydrocarbon group are replaced by deuterium, in which case it may be referred to as a "perdeuterated hydrocarbon group."

[0169] As used herein, the term "heteroalkyl" refers to an alkyl group containing heteroatoms such as N, O, S, etc. The heteroalkyl group may be substituted or unsubstituted. The heteroalkyl group may be unsaturated, saturated, branched, unbranched, cyclic, polycyclic or heterocyclic, and includes heteroalkyl, heteroalkenyl and heteroalkynyl groups. The heteroalkyl group may be fully saturated, monounsaturated or polyunsaturated, and may include divalent and polyvalent groups, for example, when it is a divalent group, it may be referred to as a "heteroalkylene group". The heteroalkyl group has a specified number of carbon atoms (i.e., C1-C 10 In some embodiments, the heteroatom contained in the heteroalkyl group can constitute the main chain of the heteroalkyl group together with the carbon atoms, such as, but not limited to, -CNC-, -COC-, -COOC, -CSC-, -CSSC and other group structures or any combination thereof. In some embodiments, the heteroatom contained in the heteroalkyl group can be a substituent attached to the carbon atom, such as, but not limited to, -C≡N, -C=N-, -CN=, -C=O, -C-OH, -C=S, -C-SH and other substitution structures. In some of the embodiments, the heteroatom contained in the heteroalkyl group can be any combination of the above-listed group structures. In some embodiments, the heteroalkyl group contains one or more, for example, 1, 2, 3, or 4, unsaturated carbon-carbon double bonds (-C=C-), carbon-carbon triple bonds (-C≡C-), -NH-, -O-, -C(O)-, -S-, -C(S)-, and / or any combination thereof. In some embodiments, the heteroalkyl group may be deuterated, also referred to as a "deuterated heteroalkyl group," i.e., one, two, three, or even more hydrogen atoms on the heteroalkyl group may be replaced by deuterium, until all hydrogen atoms on the heteroalkyl group are replaced by deuterium, in which case it may be referred to as a "perdeuterated heteroalkyl group."

[0170] As used herein, the term "alkyl" refers to a straight or branched chain saturated hydrocarbon group. i-j "alkyl" refers to an alkyl group having i to j carbon atoms. Unless otherwise specified, an alkyl group may contain 1 to 10 carbon atoms. In certain embodiments, the alkyl group contains 1 to 6 carbon atoms, such as 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n- and isopropyl, n-, sec-, iso- and tert-butyl, neopentyl, and the like. As used herein, the term "alkylene" refers to a divalent substituent that is a monovalent alkyl group having one hydrogen atom replaced by a valency.

[0171] In some embodiments, an alkyl group can be deuterated, also known as a "deuterated alkyl group."

[0172] As used herein, the term "deuterated alkyl" is a substituent obtained by replacing one or more hydrogen atoms on an alkyl group with deuterium. When an alkyl group is deuterated, one, two, three, or even more hydrogen atoms on the alkyl group may be replaced by deuterium, until all hydrogen atoms on the alkyl group are replaced by deuterium, in which case it may be referred to as a "perdeuterated alkyl group." In some embodiments, non-limiting examples of deuterated alkyl groups include deuterated methyl groups, such as monodeuterated methyl groups, dideuterated methyl groups, trideuterated methyl groups (perdeuterated methyl groups), monodeuterated ethyl groups, dideuterated ethyl groups, trideuterated ethyl groups, tetradeuterated ethyl groups, pentadeuterated ethyl groups (perdeuterated ethyl groups), and the like.

[0173] Those skilled in the art will appreciate that, in some embodiments, other types of groups mentioned herein, as long as they contain hydrogen atoms, may be deuterated. When a group is deuterated, one, two, three, or even more hydrogen atoms therein may be replaced by deuterium, until all hydrogen atoms in the group are replaced by deuterium, in which case it may be referred to as a "perdeuterated group."

[0174] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0175] The term "haloalkyl" refers to an alkyl group substituted with a halogen, wherein the halogen and alkyl are as defined above.

[0176] As used herein, term " alkenyl " refers to the straight or branched hydrocarbon radical with at least one carbon-carbon double bond, and includes the group with " cis " and " trans " orientation, or alternatively, " E " and " Z " direction.Unless otherwise indicated, alkenyl can contain 2 to 10 carbon atoms.In certain embodiments, alkenyl can contain 2 to 6 carbon atoms, for example, 2 to 5 carbon atoms, 2 to 4 carbon atoms, 2 to 3 carbon atoms.In certain embodiments, alkenyl group contains 2 carbon atoms.Non-limiting examples of alkenyl include ethylene (vinyl), propenyl, butenyl, pentenyl, 1- methyl -2- butene-1-yl, 5- hexenyl etc.

[0177] As used herein, the term "alkoxy" refers to the group -O-alkyl, wherein alkyl has the meaning defined herein.

[0178] The term "haloalkoxy" refers to an alkoxy group substituted with a halogen group, wherein halogen and alkoxy are as defined above for halogen and alkyl.

[0179] The term "heterocycloalkyl" refers to a cyclic group having a specified number of ring atoms (e.g., 3 to 10 members), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified type of heteroatom (one or more of N, O, and S), which is a monocyclic, bridged, or spirocyclic group, and each ring is saturated. A bridged ring refers to a polycyclic ring in which two or more atoms are shared between monocyclic rings. A spirocyclic ring refers to a polycyclic ring in which one atom is shared between monocyclic rings. Heterocycloalkyl includes, but is not limited to, azetidinyl, oxetane, tetrahydropyrrolyl, tetrahydrofuranyl, morpholinyl, piperidinyl, and the like; for example,

[0180] As used herein, the term "cycloalkyl" refers to non-aromatic, saturated monocyclic and polycyclic ring systems in which all ring atoms are carbon. Unless otherwise specified, a cycloalkyl group may contain 3 to 10 ring carbon atoms (i.e., C 3-10 Cycloalkyl). In certain embodiments, the cycloalkyl group can contain 3 to 9, 3 to 8, 3 to 7, 3 to 6, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 4 to 5, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6 ring-forming carbon atoms, etc. In particular, the cycloalkyl can be monocyclic or bicyclic. Alternatively, the bicyclic cycloalkyl group can include fused, spirocyclic and bridged cycloalkyl structures.

[0181] On the other hand, also include the cycloalkyl ring that wherein 1,2 or 3 heteroatoms replace into ring carbon atom.This type of group is referred to as " heterocyclic radical " or " heterocycle ", and it refers to as defined above but with at least one heteroatom selected from N, O and S as the cycloalkyl group of ring-forming atoms.Unless otherwise indicated, heterocyclic radical group can contain 3 to 10 ring-forming atoms (i.e. 3 to 10 yuan of heterocyclic radicals).In certain embodiments, heterocyclic radical group can contain 3 to 9,3 to 8,3 to 7,3 to 6,4 to 10,4 to 9,4 to 8,4 to 7,4 to 6,4 to 5,5 to 10,5 to 9,5 to 8,5 to 7,5 to 6 ring-forming atoms etc.Especially, heterocyclic radical group can be monocycle or bicyclic.Alternatively, bicyclic heterocyclic radical group can include condensation, spirocycle and bridged heterocyclic radical structure. Non-limiting examples of heterocyclyl groups include oxiranyl, pyrrolidinyl, piperidinyl, tetrahydropyranyl, piperazinyl, pyrrolidinyl, and morpholinyl. Heterocyclyl groups can also be described by using the number of carbon atoms. For example, C 3-6 Heterocyclyl refers to a heterocyclyl group containing three to six ring-forming carbon atoms, and can also contain at least one heteroatom, such as 1,2 or 3 heteroatoms as ring-forming atoms. In certain embodiments, the heterocyclyl group or heterocycle contains 1 or 2 heteroatoms as ring-forming atoms. In certain embodiments, the heterocyclyl group can be monocyclic or bicyclic, such as fused bicyclic and spiro bicyclic. In the context of the present application, the terms "heterocyclyl" and "heterocycle" can be used interchangeably.

[0182] The term "cycloalkene" refers to a cycloalkene having at least one site of unsaturation, ie, a carbon-carbon sp2 double bond (eg, a C5-C8 cycloalkene); for example,

[0183] The term "heterocyclic alkene" refers to a cyclic alkene having at least one unsaturated site, i.e., a carbon-carbon sp2 double bond, wherein at least one carbon atom is replaced by a heteroatom selected from N, O and S (e.g., 5-8 membered heterocyclic alkene, 5-6 membered heterocyclic alkene), for example,

[0184] As used herein, the term "aryl" or "aromatic ring" refers to a monocyclic, bicyclic, or polycyclic carbocyclic ring system having at least one aromatic ring. Unless otherwise indicated, an aryl group can be 6 to 10 members. In certain embodiments, an aryl group can contain 6 ring carbon atoms. All atoms within a carbocyclic aryl group are carbon atoms. Non-limiting examples of aryl groups include phenyl, naphthyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indanyl, indenyl, and the like. In the context of the present invention, the terms "aryl" and "aromatic ring" can be used interchangeably.

[0185] As used herein, the term "heteroaryl" or "heteroaromatic ring" refers to a monocyclic ring system, or a fused or bridged bicyclic ring system, wherein the ring system contains one, two, three, or four heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur; and at least one ring is an aromatic ring. Unless otherwise specified, a heteroaryl group can be 5 to 10 members. In certain embodiments, a heteroaryl group can be 5-membered or 6-membered. In certain embodiments, a heteroaryl group can contain one, two, or three heteroatoms. In certain embodiments, a heteroaryl group can contain one or two heteroatoms. The limiting examples of heteroaryl groups include benzimidazolyl, benzofuranyl, benzothiazolyl, benzothienyl, benzoxazolyl, furyl, imidazolyl, indolyl, isoindazolyl, isoquinolyl, isothiazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, purinyl, pyrrolyl, pyridyl, pyrazinyl, pyrimidinyl, quinolyl, quinolyl, thiadiazolyl, thiazolyl, thienyl, triazolyl, tetrazolyl, indolinyl, tetrahydroquinolyl, tetrahydroisoquinolyl etc.Heteroaryl groups include at least one heteroatomic ring and at least one aromatic ring with at least one as above.For example, there is at least one heteroatomic ring can be fused to one, two or three carbocyclic rings, for example aryl ring, cyclohexane ring, cyclohexene ring, cyclopentane ring, cyclopentene ring or another monocyclic heterocycle. Non-limiting examples of fused heteroaryl groups include 1,2,3,5,8,8a-hexahydroindolizine, 2,3-dihydrobenzofuran, 2,3-dihydroindole, 2,3-dihydrobenzothiophene, etc. In the context of the present invention, the terms "heteroaryl" and "heteroaromatic ring" are used interchangeably.

[0186] In one aspect, the term "aryl" may also refer to a group having a specified number of carbon atoms (e.g., C6-C 14 ) is a cyclic group consisting only of carbon atoms, which is monocyclic or polycyclic, and at least one ring is aromatic (in accordance with Huckel's rule). Aromatic rings include but are not limited to phenyl, naphthyl, or wait.

[0187] In one aspect, the term "heteroaryl" may also refer to a cyclic group having a specified number of ring atoms (e.g., 5-14 members), a specified number of heteroatoms (e.g., 1, 2, or 3), a specified heteroatom species (one, two, or three of N, O, and S), which is monocyclic or polycyclic, and at least one ring is aromatic (in accordance with Huckel's rule). A heteroaryl group is attached to the rest of the molecule via a carbon atom or a heteroatom; a heteroaryl group is attached to the rest of the molecule via a ring having heteroatoms or a ring having no heteroatoms. Heteroaryl groups include, but are not limited to, furan, pyrrole, thiophene, pyrazole, imidazole, oxazole, thiazole, pyridine, pyrimidine, indole, benzopyrrole, or wait.

[0188] As used herein, the term "oxo" refers to a divalent oxygen atom and the structure of oxo may be shown as =0.

[0189] As used herein, the term "halo" or "halogen" refers to fluoride, chloride, bromide and iodide. In certain embodiments, non-limiting examples of halo include fluoride, chloride and bromide, more particularly fluoride and chloride.

[0190] As used herein, the term "heteroatom" refers to nitrogen (N), oxygen (O), and sulfur (S), and can include any oxidized forms of nitrogen and sulfur, and any quaternized forms of basic nitrogen, unless otherwise specified.

[0191] As used herein, the term "substituted" when referring to a chemical group means that the chemical group has one or more hydrogen atoms that are removed and replaced by a substituent. As used herein, the term "substituent" has its ordinary meaning as known in the art and refers to a chemical moiety that is covalently attached to a parent group or, if appropriate, fused to a parent group. It will be understood that substitution of a given atom is limited by valence. It will be understood that a substituent may be further substituted.

[0192] When it is stated in Formula (I), Formula (III), or any embodiment thereof that a moiety is "optionally" substituted, this means that Formula (I), Formula (III), or an embodiment thereof encompasses compounds that are substituted with the indicated substituents on that moiety and compounds that do not contain the indicated substituents on that moiety (i.e., wherein the moiety is unsubstituted).

[0193] The compounds provided herein are described with reference to general formulae and specific compounds. In addition, the compounds of the present application may exist in a variety of different forms or derivatives, all of which are within the scope of the present invention. These include, for example, pharmaceutically acceptable salts, etc.

[0194] As used herein, unless otherwise indicated, the term "pharmaceutically acceptable salt" includes salts that maintain the biological effectiveness of the free acid / alkaline form of a particular compound and are not undesirable in biology or other aspects. Pharmaceutically acceptable salts can include salts formed with inorganic bases or acids and organic bases or acids. In the case where the compound of the present application contains one or more acidic or basic groups, the present invention also includes their corresponding pharmaceutically acceptable salts. Therefore, the compound of the present application containing an acidic group (such as a carboxyl group) can exist in salt form and can be used according to the present invention, for example, alkali metal salts, alkaline earth metal salts, aluminum salts or ammonium salts. More non-limiting examples of such salts include lithium salts, sodium salts, potassium salts, calcium salts, magnesium salts, barium salts or salts with ammonia or organic amines (such as ethylamine, ethanolamine, diethanolamine, triethanolamine, piperidine, N-methylglutamine or amino acids). For example, by reacting a compound with an acidic group with a suitable base (such as lithium hydroxide, sodium hydroxide, sodium propoxide, potassium hydroxide, potassium ethoxide, magnesium hydroxide, calcium hydroxide or barium hydroxide), these salts are easily obtained. Other alkali salts of compound of the present application include but are not limited to copper (I), copper (II), iron (II), iron (III), manganese (II) and zinc salt.Compound of the present application contains one or more basic groups, such as can be protonated group, can exist in the form of salt, and can be used according to the present invention with the form of addition salt of them and inorganic acid or organic acid.The example of suitable acid includes hydrogen chloride, hydrogen bromide, hydrogen iodide, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-methyl benzenesulfonic acid, naphthalenedisulfonic acid, sulfoacetic acid, trifluoroacetic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, carbonic acid, formic acid, propionic acid, pivalic acid, diethyl acetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, malonic acid, maleic acid, malic acid, pamoic acid, mandelic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, taurocholic acid, glutaric acid, stearic acid, glutamic acid or aspartic acid, and other acids well known by persons skilled in the art. The salt formed is especially hydrochloride, chloride, hydrobromide, bromide, iodide, sulfate, phosphate, methylsulfonate (mesylate), toluenesulfonate, carbonate, bicarbonate, formates, acetate, sulfoacetate, trifluoromethanesulfonate, oxalate, malonate, maleate, succinate, tartrate, malate, pamoate, mandelate, fumarate, lactate, citrate, glutarate, stearate, aspartate and glutamate. In addition, the stoichiometry of the salt formed by the compound of the application can be an integer multiple or non-integer multiple of 1.

[0195] The compounds of the present application containing basic nitrogen groups can be treated with reagents such as C 1-4 Quaternization of alkyl halides, for example, methyl, ethyl, isopropyl and tert-butyl chloride, bromine and iodine; di-C 1-4 Alkyl sulfates, such as dimethyl sulfate, diethyl sulfate, and dipentyl sulfate; C 10-18 Alkyl halides, such as decyl, dodecyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides; and aryl C 1-4 Alkyl halides, such as benzyl chloride and phenethyl bromide.

[0196] If the compound of the present application contains both an acidic group and a basic group in the molecule, the present invention also includes, in addition to the above-mentioned salt forms, inner salts or betaines (zwitterions). The corresponding salts can be obtained by conventional methods known to those skilled in the art, for example, by contacting them with organic or inorganic acids or bases in a solvent or dispersant, or by anion exchange or cation exchange with other salts. The present invention also includes all salts of the compounds of the present application, which are not directly suitable for use in medicines due to their low physiological compatibility, but can be used, for example, as intermediates in chemical reactions or for the preparation of pharmaceutically acceptable salts. For a review of more suitable salts, see Stahl and Wermuth, Handbook of Pharmaceutical Salts: Properties, Selection and Use (Wiley-VCH, 2002).

[0197] The compound of formula (I), the compound of formula (III) and pharmaceutically acceptable salts thereof may exist in unsolvated and solvated forms.

[0198] Compounds of formula (I) or (III) may have one or more chiral (asymmetric) centers. All stereoisomeric forms of compounds of formula (I) or (III) are encompassed by the present invention. Asymmetric centers present in compounds of formula (I) or (III) may have (R) or (S) configurations independently of one another. When the bond of a chiral carbon is described as a straight line in the structural formula of the present invention, or when the compound name is described in the absence of (R) or (S) chiral names of chiral carbons, it is understood that (R) and (S) configurations of each such chiral carbon and therefore each enantiomer or diastereomer and mixtures thereof are included in the formula or name. The generation of a specific stereoisomer or mixture thereof can be identified in the example of obtaining such stereoisomers or mixtures, but this in no way limits all stereoisomers and mixtures thereof to being included within the scope of the present invention. When the bond of a chiral carbon is depicted as a triangular solid line or a dashed line in the structural formula of the present invention, or when the name of a compound is depicted with a (R) or (S) chiral designation of a chiral carbon, it should be understood that the compound represented by the structural formula or name has a definite stereo configuration at the chiral carbon position and will be distinguished from other stereoisomers, enantiomers, diastereomers, or mixtures thereof.

[0199] The present invention includes all possible enantiomers and diastereomers and mixtures of two or more stereoisomers, for example, mixtures of enantiomers and / or diastereomers of all ratios. Therefore, enantiomers are enantiomerically pure forms (as left-handed and right-handed enantiomers), racemic forms and two enantiomers of the present invention's subject matter in the form of mixtures of all ratios. In the case of cis / trans isomers, the present invention includes mixtures of all ratios of cis-form and trans-form and these forms. If necessary, single stereoisomers can be prepared by conventional methods (for example, by chromatography or crystallization, by using stereochemically uniform synthesis starting materials or by stereoselective synthesis). Optionally, derivatization can be carried out before stereoisomer separation. The separation of stereoisomer mixtures can be carried out in an intermediate step during the synthesis of formula (I) compound or formula (III) compound, or can be carried out on final racemic product. Absolute stereochemistry can be determined by X-ray crystallography of crystalline products or crystalline intermediates which have been derivatized, if necessary, with reagents containing stereocenters of known configuration. Alternatively, absolute stereochemistry can be determined by vibrational circular dichroism (VCD) spectroscopy.

[0200] When describing the structures of compounds herein, the undesignated hydrogen atoms in the shown formula are D (i.e., 2 H), it is generally understood that the hydrogen at this position is a hydrogen isotope " 1 hydrogen( 1H)" or in a form with natural isotopic abundance in its natural state. The structures shown indicate that the hydrogen atoms are D (i.e., 2 H, deuterium), it should be understood that the hydrogen at this position is a hydrogen isotope " 2 hydrogen( 2 H, D, deuterium)" or in a form in which deuterium is present at a greater isotopic abundance than natural deuterium (e.g., a deuterium abundance of greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, greater than 95%, greater than 96%, greater than 97%, greater than 98%, greater than 99%, greater than 99.5%, or 100%).

[0201] Use and application

[0202] The compounds of the present application (compounds of formula (I) or formula (III)) or pharmaceutically acceptable salts thereof, including mixtures thereof in all ratios, can be used as pharmaceuticals. They exhibit pharmacological activity that inhibits the COPI / Arf1-lipolysis β-oxidation pathway. Utilizing this activity, the compounds of the present application can kill progenitor cells, stem cells, cancer stem cells, or cancer cells and induce anti-tumor immune responses. They can be used to treat conditions or diseases associated with Arf1 pathway activity; treat refractory, recurrent, or metastatic cancers; and selectively kill cancer cells using specific dosing regimens.

[0203] Therefore, the compounds of the present application are particularly suitable as Arf1 inhibitors for the treatment of diseases and conditions associated with the Arf1 pathway, particularly the COPI / Arf1-lipolysis β-oxidation pathway, such as cancer, including but not limited to the following: breast cancer, head and neck cancer, lung cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, renal cell carcinoma, melanoma, hepatocellular carcinoma, cervical cancer, sarcoma, brain tumor, gastric cancer, multiple myeloma, leukemia and lymphoma, etc.

[0204] The compound of the present application can be used in an amount effective to treat a disease or condition as described herein. The compound of the present application can be used as the compound itself, or alternatively, as a pharmaceutically acceptable salt. For the purpose of administration and administration, the compound of the present application itself (compound of formula (I) or formula (III) compound) or its pharmaceutically acceptable salt is referred to as the compound of the present application or the compound of the present disclosure.

[0205] The compounds of the present application are administered by any suitable route in the form of pharmaceutical compositions suitable for such routes and at a dose effective for the intended treatment. The compounds of the present application can be administered orally, rectally, vaginally, parenterally or topically.

[0206] As used herein, the term "administer" refers to absorbing, ingesting, injecting, inhaling, implanting or otherwise introducing a compound of the present application or its pharmaceutical composition. The term "treating" refers to reversing, alleviating, delaying the onset of "pathological conditions" (e.g., diseases, disorders or conditions, or one or more signs or symptoms thereof) as described herein or inhibiting their progression. In certain embodiments, treatment may be administered after one or more signs or symptoms of the disease or condition have developed or have been observed. In other embodiments, treatment may be performed in the absence of signs or symptoms of the disease or condition. For example, susceptible individuals may be treated before the onset of symptoms (e.g., according to a history of symptoms and / or according to genetic or other susceptibility factors). Treatment may also be continued after the symptoms subside, for example, to delay or prevent recurrence. As used herein, the terms "disease," "disorder," "condition," and "pathological condition" are used interchangeably.

[0207] Those skilled in the art can determine the dosage level to be administered by routine experiments. The dosage regimen of the compound of the present application and / or the composition comprising the compound is based on a variety of factors, including the type, age, weight, sex and medical condition of the patient; the severity of the condition; the route of administration; and the activity of the specific compound used. Therefore, the dosage regimen can be very different. For example, the dosage level of the compound of the present application can be about 0.001 to about 100 mg / kg (i.e., mg / kg body weight) per day. In certain embodiments, the total daily dose of the compound of the present application administered in a single or divided dose can be about 0.001 to about 10 mg / kg. It is not uncommon for the administration of the compound of the present application to be repeated multiple times in one day.

[0208] In some embodiments, the compounds of the present disclosure may be used in combination with one or more other therapeutic agents. In some embodiments, non-limiting examples of such other therapeutic agents include immune checkpoint inhibitors, such as anti-PD-1 antibodies, etc. These therapeutic agents may be administered before, after, or simultaneously with the administration of the compounds of the present disclosure.

[0209] As used herein, the term "immune checkpoint inhibitor" refers to a molecule that fully or partially reduces, inhibits, interferes with, or modulates one or more immune checkpoint proteins.

[0210] As used herein, the term "immune checkpoint protein" has its general meaning in this area, and refers to molecules expressed by T cells, wherein or open signal (stimulatory checkpoint molecules) or weaken signal (inhibitory checkpoint molecules). Immune checkpoint molecules are well known in the art, to constitute immune checkpoint pathways similar to CTLA-4 and PD-1 dependent pathways (see, for example, Pardoll, 2012.Nature Rev Cancer 12: 252-264; Mellman et al., 2011.Nature 480: 480-489). The example of stimulatory checkpoints includes CD27, CD28, CD40, CD122, CD137, OX40, GITR and ICOS. The example of inhibitory checkpoint molecules includes A2AR, B7-H3, B7-H4, BTLA, CTLA-4, CD277, IDO, KIR, PD-1, LAG-3, TIM-3 and VISTA. PD-1, the programmed death 1 (PD-1) receptor, has two ligands, PD-L1 and PD-L2. One advantage of targeting PD-1 is that it can restore immune function within the tumor microenvironment. Tumor cells often exploit these checkpoints to evade detection by the immune system. Therefore, inhibiting checkpoint proteins on the immune system can enhance anti-tumor T cell responses.

[0211] In some embodiments, an immune checkpoint inhibitor refers to any compound that inhibits the function of an immune checkpoint protein. Inhibition includes both reduced function and complete blockade. In some embodiments, an immune checkpoint inhibitor can be an antibody, synthetic or natural sequence peptide, small molecule, or aptamer that binds to an immune checkpoint protein and its ligand.

[0212] In some embodiments, the immune checkpoint inhibitor is an antibody. Typically, the antibody is directed against A2AR, B7-H3, B7-H4, BTLA, CTLA-4, CD277, IDO, KIR, PD-1, LAG-3, TIM-3, or VISTA.

[0213] In some embodiments, the immune checkpoint inhibitor is an anti-PD-1 antibody. Examples of commercialized anti-PD-1 antibodies include nivolumab (BMS) and pembrolizumab (also known as lambrolizumab, or MK-3475, MERCK).

[0214] Pharmaceutical composition

[0215] In some aspects, the present disclosure relates to pharmaceutical compositions comprising a compound of Formula (I) or a compound of Formula (III), a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient as provided herein.

[0216] As used herein, the term "pharmaceutically acceptable carrier and / or excipient" refers to a carrier or excipient that can be used to prepare a pharmaceutical composition, which is generally safe, non-toxic, and not biologically or otherwise undesirable, and includes carriers or excipients that are acceptable for veterinary use as well as human pharmaceutical use. As used herein, a pharmaceutically acceptable carrier or excipient includes one or more such carriers or excipients. The specific carrier or excipient used will depend on the manner and purpose of using the compound of the present application. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, for example, Ansel, Howard C et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R. et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. One or more of buffers, stabilizers, surfactants, wetting agents, lubricants, emulsifiers, suspending agents, preservatives, antioxidants, opacifiers, glidants, processing aids, colorants, sweeteners, flavorings, flavorings, diluents and other known additives may also be included to provide a refined presentation of the drug (i.e., the compound or pharmaceutical composition provided herein) or to aid in the production of a pharmaceutical product (i.e., the drug).

[0217] The compositions of the present invention can be formulated into a variety of forms. These include, for example, liquid, semisolid and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes, suppositories, etc. The form depends on the intended mode of administration and therapeutic application.

[0218] The pharmaceutical compositions of the present invention can be prepared by any well-known pharmaceutical techniques (e.g., effective formulations and administration procedures). The above considerations regarding effective formulations and administration procedures are well known in the art and are described in standard textbooks. For example, the formulation of drug products is discussed in Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania, 1975; Liberman et al., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Kibbe et al., eds., Handbook of Pharmaceutical Excipients, 3rd ed., American Pharmaceutical Association, Washington, 1999.

[0219] In another aspect, the present disclosure relates to a kit for treating diseases and conditions associated with Arf1 pathway activity, such as cancer, comprising a compound of Formula (I) or Formula (III), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, a container, and optionally a package insert or label indicating treatment or prevention.

[0220] Treatment

[0221] In another aspect, the present disclosure relates to a method for treating diseases and conditions associated with Arf1 pathway activity in a subject in need thereof, such as a method for treating cancer, wherein the compound of the present application has Arf1 inhibitory activity, and the method comprises administering to the subject a therapeutically effective amount of a compound of formula (I) or formula (III) as provided herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above.

[0222] As used herein, the term "subject in need thereof" refers to a subject who has a disease or condition associated with Arf1 pathway activity, such as cancer, or a subject who is at increased risk of developing a disease or condition associated with Arf1 pathway activity relative to the population as a whole. In certain embodiments, the subject is a warm-blooded animal. In certain embodiments, the warm-blooded animal is a mammal. In certain embodiments, the warm-blooded animal is a human.

[0223] As used herein, the term "diseases and conditions associated with Arf1 pathway activity" refers to any pathophysiological condition in which inhibition of Arf1 would be beneficial. In certain embodiments, the diseases and conditions associated with Arf1 pathway activity are cancers. In certain embodiments, the diseases and conditions associated with Arf1 pathway activity are cancers or tumors selected from the group consisting of breast cancer, head and neck cancer, lung cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, renal cell carcinoma, melanoma, hepatocellular carcinoma, cervical cancer, sarcoma, brain tumor, gastric cancer, multiple myeloma, leukemia, and lymphoma. In certain embodiments, the disease associated with Arf1 pathway activity is selected from the group consisting of autoimmune diseases, inflammatory diseases, inflammatory bowel disease, arthritis, autoimmune demyelinating disorders, Alzheimer's disease, amyotrophic lateral sclerosis, stroke, ischemia-reperfusion injury, multiple sclerosis, and other neurodegenerative diseases.

[0224] In yet another aspect, the present disclosure relates to compounds of Formula (I) or Formula (III), their pharmaceutically acceptable salts, or pharmaceutical compositions as described herein for use in treating diseases and conditions associated with Arf1 pathway activity, such as cancer or tumors.

[0225] In another aspect, the present disclosure relates to the use of a compound of formula (I) or formula (III), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, as provided herein, in the preparation of a medicament for treating diseases and conditions associated with Arf1 pathway activity, such as cancer or tumors.

[0226] The compounds of formula (I) or (III) as provided herein, their pharmaceutically acceptable salts, or the pharmaceutical compositions as described above increase the expression of MHC-I and MHC-II, or the compounds of formula (I) or (III) as provided herein, their pharmaceutically acceptable salts, or the pharmaceutical compositions as described above enhance the infiltration and activation of T cells into tumors compared to DMSO; or the compounds of formula (I) or (III) as provided herein, their pharmaceutically acceptable salts, or the pharmaceutical compositions as described above increase the expression of T cell activation markers, such as GzmA, GzmB, and Perforin.

[0227] The compounds of formula (I) or formula (III), their pharmaceutically acceptable salts, or the pharmaceutical compositions described above as provided herein increase the expression of at least one inflammatory cytokine or chemokine among IFNγ, IL-1β, CCL5, CXCL10, CXCL11, and CCL22.

[0228] The compounds of formula (I) or formula (III), their pharmaceutically acceptable salts, or the pharmaceutical compositions described above as provided herein increase the expression of at least one inflammatory cytokine or chemokine among IFNγ, IL-1β, CCL5, CXCL10, CXCL11, and CCL22.

[0229] Detection and diagnosis

[0230] In yet another aspect, the present disclosure relates to a compound of formula (I) or (III), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as provided herein for use as a test reagent for diagnosing a disease associated with Arf1 pathway activity in a subject.

[0231] synthesis

[0232] The compounds of the present invention can be prepared by the general and specific methods described below using the common knowledge of those skilled in the art of synthetic organic chemistry. Such common knowledge can be found in standard reference books, for example, Barton and Ollis (eds.), Comprehensive Organic Chemistry, Elsevier; Richard Larock, Comprehensive Organic Transformations: A Guide to Functional Group Preparation, John Wiley and Sons; and Compendium of Organic Synthesis Methods, Volumes I-XII, Wiley-Interscience.

[0233] The schemes described below are intended to provide general methods for preparing the compounds of the present application.

[0234] Some compounds of the present disclosure may contain single or multiple chiral centers with stereochemical designations (R) or (S). It will be apparent to those skilled in the art that all synthetic transformations can be performed in a similar manner, regardless of whether the material is enantiomerically enriched or racemic. In addition, the resolution of optically active materials can be performed at any desired point in the experiment using well-known methods, such as those described herein and in the chemical literature.

[0235] Example

[0236] To describe the present invention in more detail, the following examples are provided. The examples described herein are used to illustrate the compounds, methods, and compositions provided herein and their applications as Arf1 inhibitors and in anti-tumor applications, and should not be interpreted in any way as limiting their scope.

[0237] During the synthesis process, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules involved. This can be achieved by conventional protecting groups, such as those described in TW Greene and PGM Wutts, Protective Groups in Organic Synthesis, 4th Edition, John Wiley and Sons. The protecting groups are optionally removed at a convenient subsequent stage using methods well known in the art.

[0238] The compounds of the present application can be easily prepared using readily available starting materials, reagents and conventional synthesis procedures according to the reaction schemes familiar to those skilled in the art or their modifications. In these reactions, variants known to those skilled in the art but not mentioned in more detail can also be used. In addition, according to the reaction schemes and embodiments described herein, other methods for preparing the compounds of the present application will be apparent to those skilled in the art. Unless otherwise indicated, all variables are as defined above. In general, in chemical procedures, all reagents and starting materials can be purchased from commercial suppliers or can be easily prepared by those skilled in the art.

[0239] The compounds used in the examples are summarized in Table 1 below.

[0240] Example 1

[0241] Synthesis of compound 1

[0242] Step 1: General method for the preparation of compound 1-2 series

[0243] Compound 1-1 (1 eq), arylboronic acid (1.1 eq), potassium carbonate (1.2 eq) and PdCl2(dppf) (0.1 eq) were dissolved in a THF / H2O mixed solvent (v / v = 4 / 1). The reaction solution was replaced with nitrogen three times, then heated to 65°C and stirred for 16 h. LCMS or TLC analysis showed that compound 1-1 was substantially completely converted. The reaction solution was cooled to room temperature, concentrated under reduced pressure, extracted three times with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a residue, which was separated and purified by column chromatography (silica gel, petroleum ether / ethyl acetate = 20 / 1 to 10 / 1) to obtain compound 1-2.

[0244] Steps 2 and 3: General method for the preparation of compound 1 series

[0245] Compound 1-2 (1.0 eq) was dissolved in MeOH, and 85% hydrazine hydrate (1.5 eq) and triethylamine (1.5 eq) were added. The reaction mixture was heated to 50°C and stirred for 4 h. LCMS or TLC analysis revealed that compound 1-2 was essentially completely converted. The reaction mixture was concentrated under reduced pressure, and the residue was washed three times with water, then dissolved in methanol. 5-Indolecarboxaldehyde (1.2 eq) was added, and the final mixture was allowed to react at room temperature for 4 h. The reaction was complete as determined by LCMS or TLC. The reaction mixture was filtered, and the filter cake was collected and recrystallized from methanol or a DMSO / H2O mixture to obtain the target compound 1.

[0246] Compounds 2 to 54 were synthesized by using the synthesis method of compound 1 and using the corresponding arylboronic acid or alkylboronic acid instead of phenylboronic acid.

[0247] Table 1. Example compound information

[0248] Example 2

[0249] Synthesis of compound 55

[0250] Step 1: General method for the preparation of compound 55-2 series

[0251] Compound 55-1 (1.0 eq) was dissolved in MeOH, and formaldehyde aqueous solution (1.2 eq) was added. The reaction solution was stirred at room temperature for 30 minutes, and then sodium cyanoborohydride (1.5 eq) was added. The mixture was stirred at room temperature for 8 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was separated and purified by column chromatography (silica gel, dichloromethane / methanol = 40 / 1 to 10 / 1) to obtain the target compound 55-2.

[0252] Steps 2 and 3: General method for the preparation of compound 55 series

[0253] Compound 55-2 (1.0 eq) was dissolved in MeOH, and 85% hydrazine hydrate (1.5 eq) and triethylamine (1.5 eq) were added. The reaction mixture was heated to 50°C and stirred for 4 h. LCMS or TLC analysis revealed that compound 55-2 was essentially completely converted. The reaction mixture was concentrated under reduced pressure, and the residue was washed three times with water, then dissolved in methanol. 5-Indolecarboxaldehyde (1.2 eq) was added, and the final mixture was allowed to react at room temperature for 4 h. The reaction was complete as determined by LCMS or TLC. The reaction mixture was filtered, and the filter cake was collected and recrystallized from methanol or a DMSO / H2O mixture to obtain the target compound 55.

[0254] Compounds 56 to 58 were synthesized by using the synthetic method of compound 55 and replacing formaldehyde with the corresponding alkyl aldehyde or aryl aldehyde.

[0255] Table 2. Example compound information

[0256] Example 3

[0257] Synthesis of compound 59

[0258] Step 1: General method for the preparation of compound 59-2 series

[0259] Compound 55-1 (1.0 eq) was dissolved in DCM, and triethylamine (3.0 eq) and p-fluorobenzoyl chloride (1.2 eq) were added. The reaction solution was stirred at room temperature for 4 h. LCMS or TLC detected that the reaction was complete. Water was added to the reaction system, and the mixture was extracted three times with DCM. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a residue. The residue was separated and purified by column chromatography (silica gel, petroleum ether / ethyl acetate) to obtain the target compound 59-2.

[0260] Steps 2 and 3: General method for the preparation of compound 55 series

[0261] Compound 59-2 (1.0 eq) was dissolved in MeOH, and 85% hydrazine hydrate (1.5 eq) and triethylamine (1.5 eq) were added. The reaction mixture was heated to 50°C and stirred for 4 h. LCMS or TLC analysis revealed that compound 59-2 was essentially completely converted. The reaction mixture was concentrated under reduced pressure, and the residue was washed three times with water, then dissolved in methanol. 5-Indolecarboxaldehyde (1.2 eq) was added, and the final mixture was allowed to react at room temperature for 4 h. The reaction was complete as determined by LCMS or TLC. The reaction mixture was filtered, and the filter cake was collected and recrystallized from methanol or a DMSO / H2O mixture to obtain the target compound 59.

[0262] Compounds 60 to 65 were synthesized by using the synthetic method of compound 59 and replacing p-fluorobenzoyl chloride with the corresponding arylcarbonyl chloride or alkylcarbonyl chloride.

[0263] Table 3. Example compound information

[0264] Effect Example 1. Solubility of the compounds of the present invention in DMSO and H2O.

[0265] In this example, approximately 10 mg of a finely powdered compound (measured using an electronic balance) was placed in a volume of solvent at room temperature (the volume was gradually increased in 1 μL increments). The solution was shaken vigorously for 30 seconds every 5 minutes, and the solution was observed for 30 minutes. Complete dissolution was considered complete if no visible solute particles were present. This method was used to measure the solubility of the compounds synthesized in the Preparation Examples in DMSO and HO, and the results are shown in Table 4. The results demonstrate that the compounds of the present invention exhibit high solubility in DMSO, and some also exhibit high solubility in water, surpassing the control compound DU101.

[0266] Table 4. Solubility of the compounds of the present invention in DMSO and H2O

[0267] ND: Indicates not tested.

[0268] Effect Example 2. Cytotoxicity study of the compounds of the present invention.

[0269] CCK8-assay for cytotoxicity IC 50 In a 96-well plate, the amount of cells per well was determined based on cell size and proliferation rate. 100 μL of CT-26 cell suspension (1×10 3 / well), cultured at 37 ° C, 5% CO2 for 24 hours. The next day, the culture medium was discarded, and 100 μL of culture medium containing different concentrations (0.01, 0.1, 1.0, 10.0, 100.0 μM) of the drug to be tested was added to the well plate. Incubated in a 37 ° C incubator for 24 hours, 48 ​​hours, and 96 hours. Along the side walls of the wells, 10 μL of CCK-8 solution (Shanghai Biyuntian Biotechnology Co., Ltd. Product No. C0038) was added to each well, and the culture plate was placed in a 37 ° C incubator and incubated for 4 hours. The absorbance at 450 nm was measured using an enzyme marker. The cytotoxicity results are shown in Table 5, which indicate that the novel Arf1 inhibitor obtained by the present invention has low cytotoxicity and good safety.

[0270] Table 5. Cytotoxicity of the compounds of the present invention

[0271] N / A means not tested.

[0272] Example 3. Inhibition assay of Arf1 activation.

[0273] In this example, the Arf1 inhibitors DU101 or DU102 reported (Wang et al., 2023) were used. As a positive control, the compounds synthesized in the above examples were tested for their ability to inhibit Arf1 activation. Activated Arf1 was detected using the Pierce Active Arf1 Pull-Down and Detection Kit (Thermo Scientific, Cat#16121) with Huh-7 liver cancer cell lysate. This comprehensive kit selectively enriches and detects GTP-bound Arf1 GTPase through specific protein interactions with the GGA3 protein-binding domain.

[0274] The test results, shown in Figures 1-3, demonstrate that the compounds of the present application are effective inhibitors of Arf1 activation. Compared with the control drug DU101, compounds 1, 3, 7, 17, 19, 24, 27, 43, and 44 have stronger abilities to inhibit Arf1 activation in Huh-7 cells. Compared with the control drug DU102, compound 55 is more effective in inhibiting Arf1 activation in Huh-7 cells.

[0275] Example 4. Inhibition of Drosophila tumor growth.

[0276] In this example, the reported Arf1 inhibitor DU101 was used as a positive control to test the inhibitory effect of the compounds synthesized in the above examples on the growth of orthotopic tumors in Drosophila.

[0277] As previously described, the Drosophila kidney stem cells (Ras V12 -PMML) clones expressing Ras induced the generation of renal stem cell tumor clusters (see Extended Data Fig. 4 in Singh et al., Nature. 2016 10, 06; 538(7623): 109–113). V12 -PMML cloned flies were cultured for 4 days to allow tumor growth, then switched to food containing the indicated drug for another 4 days, and then given a Ras V12 Flies fed with PMML were fed normal food containing DMSO or 5 μM of the compound. In the indicated groups, 5 × 10 3 μm 2The area of ​​esg>GFP+ tumor cells is only expressed in Drosophila intestinal stem cells and enteroblasts and can be used as a marker gene to characterize tumor size.

[0278] The test results are shown in Figures 4-8 and Table 6 (using T-test statistics, where * represents statistically significant differences, i.e., * represents P < 0.05, ** represents P < 0.01, and *** represents P < 0.001), indicating that the compounds of the present application can effectively inhibit the growth of tumors in Drosophila.

[0279] Compared with the control compound DU102 or DU101, compounds 7, 8, 18, 19, 21, 28, 35, 49, 50, 51, 55 and 64 of the present application have stronger in vivo anti-tumor effects.

[0280] Table 6. Activity of the compounds of the present invention in inhibiting tumor growth in Drosophila

[0281] Example 5. Inhibition of CT26 colon cancer in BALB / c mice.

[0282] In this example, the reported Arf1 inhibitor DU102 was used as a positive control to test the inhibitory effect of compound 55 synthesized in the above example on the growth of colon cancer.

[0283] CT26 cells were inoculated into the left or right abdomen of 6-week-old male BALB / c mice (Jiangsu Jicui Yaokang Biotechnology Co., Ltd.) at a dose of 5×10 5 cells / mouse. The mice were divided into two groups: a control group and a compound 55 treatment group. After about 7 days, the mice were given ddH2O or 5 mg / kg compound 55 (dissolved in ddH2O) daily by gavage for two consecutive weeks. The tumor volume was measured every 2-3 days using a digital caliper (ULINE, Cat#H-7352), and then calculated using the formula 1 / 2×longitudinal diameter (length)×maximum transverse diameter (width). When the maximum tumor volume reached 2000mm 3 When , the mice were euthanized.

[0284] The test results are shown in Figures 9-10 (wherein, T-test statistics are used, * represents statistically significant difference, i.e., P < 0.05; the number of “·” represents the number of samples), indicating that compound 55 of the present application can effectively inhibit CT26 colon cancer in BALB / c mice.

[0285] The foregoing description is intended to be merely illustrative of the principles of the invention. Furthermore, since many modifications and variations will be apparent to those skilled in the art, it is not intended to limit the invention to the exact configuration and process described above. Therefore, all suitable modifications and equivalents are intended to fall within the scope of the invention as defined by the appended claims.

[0286] All publications, patents, and patent applications cited herein are hereby incorporated by reference into this disclosure in their entirety.

Claims

1. A compound of formula (III) or a pharmaceutically acceptable salt thereof, characterized in that, Among them, represents a double bond, which is in the Z-form, E-form or a mixture of Z-form and E-form; X3, X4, X5, X6, and X9 are each independently C(R x ), or N, and R x is independently H, D, CN, OH, NH2, NHSO2C 1- 6-alkyl, halogen, C 1-6 -alkyl, C 1-6 -haloalkyl, C 1-6 -alkoxy, or C 1-6 -haloalkoxy; X7 and X8 are each independently C(R x ), or N, and R x is independently H, D, CN, OH, NH2, NHSO2C 1-6 alkyl, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy or C 1-6 haloalkoxy; Alternatively, X7 is C(R x ), X8 is C(R x ), R in X7 x and R in X8 x together with the atoms to which they are attached form an optionally 1-, 2- or 3-R e substituted ring D, said ring D being a C5-C6 cycloalkene, 5-6 membered heteroalkene, benzene ring or 5-6 membered heteroaromatic ring, and in the 5-6 membered heteroalkene and 5-6 membered heteroaromatic ring, the heteroatoms are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3; Ring A is a C5-C6 cycloalkene, 5-6 membered heteroalkene, benzene ring or 5-6 membered heteroaromatic ring. In the 5-6 membered heteroalkene and 5-6 membered heteroaromatic ring, the heteroatoms are independently selected from one or more of N, O and S, and the number of heteroatoms is independently 1, 2 or 3; n is 1, 2 or 3; Each R1 is independently H, D, CN, OH, halogen, biphenyl, -C(O)-OC 1-6 alkyl, optionally substituted with one, two or three R a alkyl, optionally substituted with one, two or three R 1-6 alkoxy, optionally substituted with one, two or three R b alkyl, optionally substituted with one, two or three R 1-6 cycloalkyl, optionally substituted with one, two or three R a alkyl, optionally substituted with one, two or three R 3-10 3- to 10-membered heteroalkyl, optionally substituted with one, two or three R a aryl or 5- to 14-membered heteroaryl, optionally substituted with one, two or three R c alkyl, optionally substituted with one, two or three R 6-14 wherein, in the 5- to 14-membered heteroaryl, the heteroatoms are independently selected from one or more of N, O and S, and the number of heteroatoms is independently one, two or three; c ​ or R1 on two adjacent carbons and the carbon atoms to which they are attached together form an optionally 1-, 2- or 3-R d substituted C 5-8 cycloalkene or an optionally 1-, 2- or 3-R d substituted 5- to 8-membered heteroalkene, wherein in the 5- to 8-membered heteroalkene, the heteroatoms are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; R a 、R b 、R c 、R d and R e are each independently D, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1- 6-haloalkoxy, -C(O)-C 1-6 alkyl, -C(O)-OC 1-6 alkyl, -C(O)-C 3-6 cycloalkyl, oxo(=O), -NH2, optionally substituted by 1, 2 or 3 R a-1 substituted -C(O)-C 6-14 aryl or optionally 1, 2 or 3 R a-1 substituted -C(O)-5- to 14-membered heteroaryl; R a-1 each independently represents a halogen; The compound of formula (I) is not 2. A compound of formula (I) or a pharmaceutically acceptable salt thereof, characterized in that, wherein, X1 and X2 are selected from C, N, O or S; represents a single bond or a double bond; represents a double bond, which can be in the Z or E form; X3, X4, X5, X6, X7, X8 and X9 are each independently selected from C(R x ), or N, where R x is independently selected from the group consisting of H, D, CN, OH, NH2, NHSO2Me, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, and C 1-6 haloalkoxy; R1, R2, and R3 are each independently absent or selected from the group consisting of H, D, CN, OH, halogen, biphenyl, -C(O)-OC 1-6 alkyl, optionally substituted with 1 - 3 R a of C 1-6 alkyl, optionally substituted with 1 - 3 R b of C 1-6 alkoxy, optionally substituted with 1 - 3 R a of C 3- 10 cycloalkyl, optionally substituted with 1 - 3 R a of C 3-6 heterocycloalkyl, optionally substituted with 1 - 3 R c of C 6-14 aryl, optionally substituted with 1 - 3 R c of C 6-14 heteroaryl, or R1 and R2 together with the carbon atom to which they are attached form an optionally substituted C d with 1 - 3 R 5-8 carbocycle or a 5 - to 8 - membered heterocycle, the 5 - to 8 - membered heterocycle optionally containing one or more heteroatoms selected from N, O, and S; Ring D is absent or is optionally substituted with 1 - 3 R e C 5-6 carbocyclic ring or 5 - to 6 - membered heterocyclic ring; Each R, each time it appears a 、R b 、R c 、R d 、R e is independently selected from the group consisting of D, halogen, C 1-6 alkyl, C 2-6 alkenyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, -C(O)-C 1-6 alkyl, -C(O)-OC 1-6 alkyl, -C(O)-C 1-6 cycloalkyl, -C(O)-C 6-14 aryl, oxo(=O) and -NH2 3. The compound represented by formula (III) according to claim 1 or a pharmaceutically acceptable salt thereof, or the compound represented by formula (I) according to claim 2 or a pharmaceutically acceptable salt thereof, characterized in that, it satisfies one or more of the following conditions; (1) In the compound represented by formula (III), each of the C 1-6 alkyl and substituted C 1-6 alkyl, the C 1-6 alkyl is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl or isopentyl; for example, methyl, ethyl, tert-butyl, isopropyl, n-propyl, n-butyl or n-pentyl; preferably methyl; (2) In the compound represented by formula (III), each of the C 1-6 alkoxy group and the substituted C 1-6 alkoxy group, the C 1-6 alkoxy group is independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy or tert-butoxy; for example, methoxy or ethoxy; (3) In the compound represented by formula (III), each of the 5-6 membered heteroaryl rings and the substituted 5-6 membered heteroaryl rings is a 5-membered heteroaryl ring having 1 or 2 heteroatoms; for example, (4) In the compound represented by the formula (III), each of the 5- to 6-membered heteroalkenes is a 6-membered heteroalkene with N as the heteroatom and the number of heteroatoms being 1; for example, (5) In the compound represented by formula (III), each of the C 3-10 cycloalkyl and substituted C 3-10 in cycloalkyl, the C 3-10 cycloalkyl is independently cyclohexyl or In the compound represented by formula (III), each of the 3- to 10-membered heterocycloalkyl and the substituted 3- to 10-membered heterocycloalkyl is independently a 3- to 6-membered heterocycloalkyl; preferably a 6-membered heterocycloalkyl having O as the heteroatom and 1 heteroatom; for example, In the compound represented by formula (III), each of the C 6-14 aryl and substituted C 6-14 aryl, the C 6-14 aryl is independently C 6-10 aryl; for example, In the compound represented by the formula (III), each of the 5- to 14-membered heteroaryl groups and the substituted 5- to 14-membered heteroaryl groups is independently a 5- to 10-membered heteroaryl group; preferably a 5- to 10-membered heteroaryl group having 1 heteroatom; for example, In the compound represented by formula (III), each of the C 5-8 cycloalkene is In the compound represented by formula (III), each of the 5- to 8-membered heteroalkene and the substituted 5- to 8-membered heteroalkene is independently a 5- to 6-membered heteroalkene having N as the heteroatom and one heteroatom; for example, (11) In the compound represented by formula (III), each of the halogens is independently fluorine, chlorine, bromine or iodine; for example, fluorine; (12) In the compound represented by formula (III), each of the halo groups is independently fluoro, chloro, bromo or iodo; (13) In the compound represented by formula (III), each of the C 2-6 alkenyl and substituted C 2-6 alkenyl, the C 2-6 alkenyl is independently C 2-4 alkenyl; for example, In the compound represented by formula (I), ring A is selected from the following ring structures: In the compound represented by formula (I), R1, R2, and R3 each independently do not exist or are selected from the group consisting of H, D, CN, OH, halogen, biphenyl, -C(O)-OC 1-6 alkyl, optionally substituted with 1 - 3 R a of C 1-6 alkyl, optionally substituted with 1 - 3 R b of C 1-6 alkoxy, optionally substituted with 1 - 3 R a of C 3-10 cycloalkyl, optionally substituted with 1 - 3 R a of C 3-6 heterocycloalkyl, optionally substituted with 1 - 3 R c of C 6-14 aryl and optionally substituted with 1 - 3 R c of C 6-14 heteroaryl, such as H, D, CN, OH, F, Cl, Br, I, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert - butyl, pentyl, adamantyl, cyclohexyl, tetrahydrofuranyl, methoxy, ethoxy, propoxy, butoxy, methoxycarbonyl, ethoxycarbonyl, phenyl, tolyl, ethylphenyl, methoxyphenyl, trifluoromethylphenyl, fluorophenyl, dimethylphenyl, isopropylphenyl, isobutenylphenyl, epoxybutanyl - phenyl, cyclohexyl - phenyl, biphenyl or naphthyl; In the compound represented by formula (I), represents an E-form double bond; In the compound represented by formula (I), when ring D is absent, ring C is optionally substituted with 1-5 R x ; for example, ring C is substituted with 1, 2, 3, 4 or 5 R x substituted benzene ring; or When ring D is present, ring C is optionally substituted with 1 - 3 R x ; ring C is, for example, a benzene ring substituted with 1, 2 or 3 R x ; In the compound represented by formula (I), when ring D is present, ring D is optionally substituted with 1-3 Rs e ; for example, the following ring structures optionally substituted with 1, 2 or 3 Rs e substituted: where "1" indicates the connection direction to X7 of ring C, and "2" indicates the connection direction to X8 of ring C; for example, optionally substituted by 1, 2, and / or 3 Rs e substituted and In the compound represented by formula (I), R e is C 1-4 alkyl, C 1-4 haloalkyl, C 1-4 alkoxy, C 1-4 haloalkoxy or oxo (=O), such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, trifluoromethyl, trifluoroethyl (e.g., -CH2CF3), methoxy, ethoxy, propoxy, butoxy, C 1-4 haloalkoxy or oxo (=O).

4. The compound represented by formula (III) according to claim 1 or a pharmaceutically acceptable salt thereof, or the compound represented by formula (I) according to claim 2 or a pharmaceutically acceptable salt thereof, characterized in that, it satisfies one or more of the following conditions; (1) In the compound represented by formula (III), X3 is N; (2) In the compound represented by formula (III), X4 is N; (3) In the compound represented by the formula (III), R x is independently H, OH or NHSO2Me; preferably H; (4) In the compound represented by formula (III), X5 is C(R x ); for example, CH; (5) In the compound represented by formula (III), X6 is C(R x ); for example, CH; (6) In the compound represented by formula (III), X7 is C(R x ); for example, (7) In the compound represented by formula (III), X8 is C(R x ); for example, CH; (8) In the compound represented by formula (III), X9 is C(R x ); for example, CH; (9) In the compound represented by formula (III), ring A is a 5-6 membered heteroalkene or 5-6 membered heteroaromatic ring; (10) In the compound represented by formula (III), n is 1 or 2; (11) In the compound represented by formula (III), R c is independently halogen, C 1-6 alkyl, C 2-6 alkenyl, C 1-6 haloalkyl or C 1-6 alkoxy; for example, methyl, methoxy, ethyl, trifluoromethyl, tert-butyl, fluorine, isopropyl, n-propyl or In the compound represented by the formula (III), R d is independently C 1-6 alkyl, -C(O)-5- to 14-membered heteroaryl, -C(O)-C 1- 6-alkyl, -C(O)-OC 1-6 alkyl, -C(O)-C 3-6 cycloalkyl or -C(O)-C a-1 optionally substituted by one, two or three R 6-14 aryl; for example, methyl, ethyl, n-propyl, (13) In the compound represented by formula (III), R1 is any one of the following: Case 1: Each R1 is independently H, biphenyl, -C(O)-OC 1-6 alkyl, C 1-6 alkyl, C 3-10 cycloalkyl, 3- to 10-membered heteroalkyl, optionally substituted by 1, 2 or 3 R c substituted C 6-14 aryl or optionally substituted by 1, 2 or 3 R c substituted 5- to 14-membered heteroaryl; for example, H, methyl, propyl, butyl, pentyl, ethyl Case 2: R1 on two adjacent carbons and the carbon atoms to which they are attached together form an optionally 1-, 2- or 3-R d substituted C 5-8 cycloalkene or an optionally 1-, 2- or 3-R d substituted 5- to 8-membered heterocycloalkene; for example, an optionally 1-, 2- or 3-R d substituted For another example, Preferably, R1 on two adjacent carbons and the carbon atoms to which it is attached together form a 5- to 8-membered heteroalkene optionally substituted with 1, 2, or 3 R d substituents; for example, a 5- to 8-membered heteroalkene optionally substituted with 1, 2, or 3 R d substituents Further for example In the compound represented by formula (I), the C formed by R1 and R2 together with the carbon atom to which they are attached 5-8 carbocyclic ring or 5- to 8-membered heterocyclic ring, such as selected from the following ring structures: Preferably Said C 5-8 The carbocyclic ring or 5- to 8-membered heterocyclic ring is optionally substituted with 1-3 R d ; and In the compound represented by the formula (I), when ring D does not exist, ring C is optionally substituted with 1 to 4 Rs x ; Ring C is, for example, a pyridine ring having 1, 2, 3 or 4 Rs x ; or When ring D is present, ring C is optionally substituted with 1 - 2 R x ; Ring C is, for example, a pyridine ring with 1 or 2 R x .

5. The compound represented by formula (III) according to claim 1 or a pharmaceutically acceptable salt thereof, or the compound represented by formula (I) according to claim 2 or a pharmaceutically acceptable salt thereof, characterized in that it satisfies one or more of the following conditions; (1) Among the compounds represented by the formula (III), For Among them For R1 is defined as described in any one of claims 1, 3 or 4, the For When, the R1 on two adjacent carbon atoms does not form a ring with the atoms to which they are attached; for example, (2) In the compound represented by formula (III), X7 is C(R x ), X8 is C(R x ), R x in X7 and R x in X8 together with the atoms to which they are attached form ring D, and the said ring D is a 5- or 6-membered heteroaryl ring; for example, wherein, 1 is connected to X6 and 2 is connected to X9; (3) In the compound represented by formula (I), when R1 and R2 together with the carbon atom to which they are attached form a C 5-8 carbocyclic ring or a 5- to 8-membered heterocyclic ring, the fused ring structure formed by it and ring A is selected from the following structures: Optionally, it is substituted with 1-3 R's d ; for example, In the compound represented by formula (I), ring B is selected from the following structures: For example, and (5) In the compound represented by formula (I), X5, X6, X7, X8 and X9 are any one of the following: Case 1, X5, X6, X7, X8, and X9 are each independently selected from C(R x ); Case 2, any one of X5, X6, X7, X8, and X9 is N, and the rest are C(R x ); Preferably, ring C is selected from the following ring structures: For example, 6. The compound represented by formula (III) according to claim 1 or a pharmaceutically acceptable salt thereof, or the compound represented by formula (I) according to claim 2 or a pharmaceutically acceptable salt thereof, characterized in that it satisfies one or more of the following conditions; (1) Among the compounds represented by the formula (III), For Among them For wherein, R1 is defined as described in any one of claims 1, 3 or 4, and R1 on two adjacent carbons forms a ring with the atoms to which they are attached; for example, (2) Among the compounds represented by formula (III), For For example, (3) In the compound represented by formula (I), ring A is selected from the following ring structures: In the compound represented by formula (I), rings C and D together form the following fused ring structure: Ring C is optionally substituted with 1 - 3 R's x and ring D is optionally substituted with 1 - 3 R's e ; for example, and In the compound represented by formula (I) as described in (5), the compound represented by formula (I) is 7. The compound represented by formula (III) according to claim 1 or a pharmaceutically acceptable salt thereof, or the compound represented by formula (I) according to claim 2 or a pharmaceutically acceptable salt thereof, characterized in that The compound of formula (I) or formula (III) is selected from the following compounds: Among them, represents a double bond, indicating the Z-form, E-form, or a mixture of the Z-form and E-form; preferably, is of the E form; then the compound represented by formula (I) or formula (III) is any of the following compounds:

8. A pharmaceutical composition, which comprises a compound represented by formula (III) as described in any one of claims 1, 3-7 or a pharmaceutically acceptable salt thereof, or a compound represented by formula (I) as described in any one of claims 2-7 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

9. Use of a compound of formula (III) as described in any one of claims 1, 3 - 7 or a pharmaceutically acceptable salt thereof, or a compound of formula (I) as described in any one of claims 2 - 7 or a pharmaceutically acceptable salt thereof, in the preparation of a reagent for inhibiting the activity of the intracellular Arf1 pathway in cells; wherein, The cell can be a progenitor cell, a stem cell, a cancer stem cell or a cancer cell; the reagent can induce cell death at the cellular level; the reagent can be administered to the cell in vitro or in vivo.

10. Use of a compound of formula (III) as described in any one of claims 1, 3 - 7 or a pharmaceutically acceptable salt thereof, a compound of formula (I) as described in any one of claims 2 - 7 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 8 for treating or preventing a disease associated with Arf1 pathway activity in a subject; wherein, The disease related to the Arf1 pathway activity can be cancer or tumor; the cancer or tumor can be selected from the group consisting of breast cancer, head and neck cancer, lung cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, renal cell carcinoma, melanoma, hepatocellular carcinoma, cervical cancer, sarcoma, brain tumor, gastric cancer, multiple myeloma, leukemia and lymphoma; Alternatively, the disease related to the Arf1 pathway activity is selected from the group consisting of autoimmune diseases, inflammatory diseases, inflammatory bowel disease, arthritis, autoimmune demyelinating disorders, Alzheimer's disease, amyotrophic lateral sclerosis, stroke, ischemia-reperfusion injury, multiple sclerosis and other neurodegenerative diseases; The subject can be a mammal, such as a human; Preferably, the compound represented by formula (III) or a pharmaceutically acceptable salt thereof, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition is co-administered with at least one immune checkpoint inhibitor, such as an anti-PD-1 antibody; for example, co-administered simultaneously with the anti-PD-1 antibody or administered successively in any order.

11. A kit for treating or preventing a disease associated with Arf1 pathway activity in a subject, the kit comprising the compound represented by formula (III) or a pharmaceutically acceptable salt thereof as described in any one of claims 1, 3-7, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof as described in any one of claims 2-7, or the pharmaceutical composition as described in claim 8; a container; and optionally a package insert or label indicating treatment or prevention, and the kit may further comprise at least one immune checkpoint inhibitor, such as an anti-PD-1 antibody; The disease associated with Arf1 pathway activity and the subject are as described in claim 10.

12. A kit for diagnosing a disease associated with Arf1 pathway activity in a subject, comprising at least one of the compound represented by formula (III) or a pharmaceutically acceptable salt thereof as described in any one of claims 1, 3-7, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof as described in any one of claims 2-7, or the pharmaceutical composition as described in claim 8 as a test reagent; Among them, The test reagent can detect at least one biomarker indicating the presence of a disease associated with Arf1 pathway activity; for example, the activity of Arf1 GTPase, a reporter gene for detecting lipolytic activity, detecting the formation of lipid droplets, detecting autophagy activity, or an upstream or downstream alternative regulator of Arf1 activity or function; the disease associated with Arf1 pathway activity and the subject are as described in claim 10.

13. Use of the compound represented by formula (III) or a pharmaceutically acceptable salt thereof as described in any one of claims 1, 3-7, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof as described in any one of claims 2-7, or the pharmaceutical composition as described in claim 8 in the preparation of a therapeutic vaccine for blocking tumor development; Among them, The tumors are selected from the group consisting of breast cancer, head and neck cancer, lung cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, renal cell carcinoma, melanoma, hepatocellular carcinoma, cervical cancer, sarcoma, brain tumor, gastric cancer, multiple myeloma, leukemia, and lymphoma.

14. A method for killing cells and inducing an anti-tumor immune response, the method comprising inhibiting the activity of at least one Arf1 in a cell, preferably the activity of the COPI / Arf1-lipolytic β-oxidation pathway, by an Arf1 pathway inhibitor, wherein, The cell is a progenitor cell, a stem cell, a cancer stem cell, or a cancer cell; the Arf1 pathway inhibitor is the compound represented by formula (III) or a pharmaceutically acceptable salt thereof as described in any one of claims 1, 3-7, or the compound represented by formula (I) or a pharmaceutically acceptable salt thereof as described in any one of claims 2-7.

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