USE OF 2-(2-CHLOROPHENYL)-4-[3-(DIMETHYLAMINO)PHENYL]-5-METHYL-1H-PYRAZOLO[4,3-C]PYRIDINO-3,6(2H,5H)-DIONE, AND PHARMACEUTICAL COMPOSITION

Abstract

The present invention relates to compounds, methods, compositions and uses that are capable of restoring responsiveness to immunotherapy, in particular immune checkpoint inhibitors or cancer vaccines or anti-angiogenesis treatment.

Description

51 USE OF 2-(2-CHLOROPHENYL)-4-[3-(DIMETHYLAMINO)PHENYL]-5-METHYL-1H-PYRAZOLO[4,3-C]PYRIDINO-3,6(2H,5H)-DIONE, AND PHARMACEUTICAL COMPOSITION Field of Invention

[001] The present invention relates to the use of NADPH oxidase (NOX) inhibitors, in particular NOX4 or NOX4 / 1 dual or NOX1 inhibitors, for the treatment of solid cancers in combination with cancer immunotherapy or antiVEGF treatment and related combined formulations and regimens. Fundamentals of the Invention

[002] Cancer cells face multiple cellular stresses such as hypoxia, increased metabolic demand, genomic instability, immune surveillance, lack of nutrients, changing environment after metastasis and stresses resulting from treatments such as radiotherapy, chemotherapy and targeted therapies.

[003] NADPH oxidases (NOX) are a family of enzymes that harbor the transmembrane domain 6 and transfer electrons across biological membranes. These enzymes are dedicated reactive oxygen species-generating enzymes that broadly and specifically regulate redox-sensitive signaling pathways involved in cancer development and progression, acting on specific cell membranes and microdomains through the activation of oncogenes and the inactivation of tumor suppressor proteins. NOX enzymes are considered to be an essential part of the adaptive stress response, particularly for cancer cells, thus enabling these cells to adapt and survive (Block et al., 2012, Nature Reviews, 627-637).

[004] Marked induction of NOX expression has been reported in cancer cells and host cells within the tumor environment.

[005] The interaction between the tumor microenvironment and cancer cells Petition 870250096065, dated 10 / 21 / 2025, page 17 / 90 / 51, recognizes a major role in tumor growth and metastasis. Cancer-associated fibroblasts (CAFs) are the most abundant cells found in the tumor stroma. CAFs and their transdifferentiation from fibroblast to myofibroblast lead to tumor growth and generally correlate with poor prognosis in multiple cancer types. Although CAFs promote “many of the hallmarks of neoplasia,” recent studies have highlighted a role in promoting tumor immune evasion with CAF-rich cancers that are designated as “cold immune” due to their poor therapeutic response to cancer immunotherapies such as immune checkpoint inhibitors and cancer vaccines, and their propensity to evolve to metastasis.

[006] Furthermore, the high CAF content induces a dense stroma and dense tumor microenvironment that increases interstitial fluid pressure and thus acts as a barrier to drug delivery, leading to insufficient accumulation of chemotherapies in tumors.

[007] In particular, melanoma is known as an exceptionally aggressive and treatment-resistant human cancer. Although progress has been made in the past decade, including the development of immunotherapy using immune checkpoint inhibitors, treatment for unresectable stage III, stage IV, and recurrent melanoma remains challenging with limited response rates, severe side effects, and poor prognosis. Melanoma is not driven solely by malignant melanocytes but also by altered communication between neoplastic cells and non-malignant cell populations, including fibroblasts, endothelial cells, and inflammatory cells, in the tumor stroma. CAFs remodel the extracellular matrix (ECM) and diseased tissue architecture and secrete chemical factors, which together promote the transformation process by encouraging tumor growth, angiogenesis, inflammation, and metastasis, and contribute to drug resistance. If it was Petition 870250096065, dated 10 / 21 / 2025, pp. 18 / 90 / 51 recently showed that NOX4 regulates the differentiation of myofibroblastic CAFs in multiple cancers (Hanley et al., 2018, J Natl Cancer Inst., 110), the origin of CAFs and the precise mechanisms by which CAFs contribute to cancer progression and drug resistance remain insufficiently understood. Furthermore, Hanley et al., 2018 did not suggest any specific anticancer immunotherapeutic agent as adjunctive treatment with NOX4 inhibition.

[008] Immunotherapy continues to generate interest as an effective therapeutic strategy across various types of cancer such as melanoma, non-small cell lung cancer, small cell lung cancer, head and neck cancer, renal cell carcinoma, bladder cancer, ovarian cancer, uterine endometrial cancer, uterine cervical cancer, uterine sarcoma, gastric cancer, esophageal cancer, colon cancer, hepatocellular carcinoma, breast cancer, Merkel cell carcinoma, thyroid cancer, Hodgkin lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, mycosis fungoides, peripheral T-cell lymphoma and includes various approaches, ranging from stimulating effector mechanisms to counteracting inhibitory and suppressive mechanisms.Strategies to activate effector immune cells include vaccination with tumor antigens or augmentation of antigen presentations to increase the capacity of the patient's own immune system to enhance the effectiveness of the immune response against neoplastic cells (Yaddnapudi et al., 2013, Cancer vaccines, Oncoimmunology, 2(3), e23403). Additional stimulatory strategies include adoptive cell therapy (ACT), administration of oncolytic viruses (OVs) to initiate systemic antitumor immunity, and the use of antibody-targeting members of the tumor necrosis factor receptor superfamily to enhance T cell activity. Strategies to neutralize immunosuppressive mechanisms include chemotherapy (cyclophosphamide), antibodies to decrease regulatory T cells (CD25-targeted antibodies), and antibodies. Petition 870250096065, dated 10 / 21 / 2025, page 19 / 90 / 51 against immune checkpoint molecules such as CTLA-4, PD1 and PD-L1.

[009] The field of cancer immunotherapy has recently been encouraged primarily by the approval of autologous cellular immunotherapy, sipuleucel-T, for the treatment of prostate cancer in 2010 (Topalian et al., 2011, J. Clin. Oncol., 29: 4828-36) and the approval of anti-cytotoxic T lymphocyte antibody-associated protein 4 (CTLA-4), ipilimumab, and anti-programmed cell death protein 1 (PD1) for the treatment of melanoma in 2011 and 2014 (Sharma et al., 2015, Cell, 161: 205-14).

[0010] Successful anticancer effects have been demonstrated through the use of the immune checkpoint blockade-associated cytotoxic T lymphocyte-associated protein 4 (CTLA-4) and programmed death 1 (PD-1) / PD-1 ligand (PD-L1), with the highest objective response rates observed in cancer types with a high mutational burden such as melanoma and non-small cell lung cancer (Andrews et al., 2017, Journal for ImmunoTherapy of Cancer, 25: 10). However, significant limitations exist with these therapeutic agents, with objective responses to PD-1 blockade observed in only 30–40% of patients, and most patients demonstrating innate resistance. Acquired resistance to anti-PD-1 therapy is also a problem, with approximately one-quarter of responders later demonstrating disease progression (Ribas et al., 2016, JAMA, 315: 1600–9).

[0011] Furthermore, resistance of solid tumors to anticancer treatment has also been observed for antiangiogenic therapies and has become a major concern for the use of anti-VEGF therapies (Gardner et al., 2017, Chapter 19, AntiVEGF Therapy in Cancer: A Double-Edged Sword, http: / / dx.doi.org / 10.5772 / 66763, anti-PDGF agents) since, despite their encouraging beneficial effects, patients inevitably develop resistance and frequently fail to demonstrate Petition 870250096065, dated 10 / 21 / 2025, page 20 / 90 / 51 significantly improved global survival.

[0012] Therefore, in view of recent developments in various strategies in cancer immunotherapy such as cancer vaccines, adoptive cellular immunotherapy, immune checkpoint blockade and oncolytic viruses and antiangiogenic therapies, but also the limitations encountered for their effectiveness, there is a growing need to develop efficient anticancer therapies for solid tumor cancers, in particular for cancers prone to developing resistance to immunotherapy or antiangiogenic therapies, which would allow restoring sensitivity to immunotherapy or antiangiogenic treatments or potentiating cancer vaccine treatments. Summary of the Invention

[0013] The present invention is directed towards the unexpected findings that the recently discovered ability of pharmacological inhibition of NOX4 to reverse the myofibroblastic CAF phenotype in different cancer cell lines and suppress tumor growth in multiple CAF-rich tumor models (TC1+CAF [HNSCC model], 4T1+CAF [breast cancer], MMTV-PyVT (breast cancer), MMTVHer2 / neu (breast cancer) both in vitro and / or in vivo (Hanley et al., 2018, J Natl Cancer Inst., 110) is useful for synergistically enhancing cancer immunotherapy or reversing resistance evoked by antiVEGF treatment.

[0014] The present invention is directed towards the unexpected findings that dual NOX4 / 1 inhibitors are able to restore sensitivity to immunotherapy and / or improve response to immunotherapy and antiangiogenic therapies.

[0015] The present invention is directed to compositions and methods useful for restoring responsiveness to immunotherapy, in particular for restoring responsiveness to cancer vaccines such as HPV and checkpoint blockade such as with inhibitors of Petition 870250096065, dated 10 / 21 / 2025, p. 21 / 90 / 51 PD-1 inhibitors, PD-L1 inhibitors, and CTLA-4 inhibitors.

[0016] In particular, the present invention is directed to the unexpected findings that NOX4 inhibitors are able to restore sensitivity to antitumor immunotherapy and / or improve the response to immunotherapy.

[0017] In particular, the present invention is directed towards the unexpected findings that NOX1 inhibitors are able to improve the response to antiangiogenic therapies.

[0018] The present invention is further directed to compositions and methods useful for restoring responsiveness to anti-angiogenic therapies, in particular for restoring responsiveness to anti-VEGF treatment and / or reducing or preventing the appearance of resistance to anti-VEGF treatment.

[0019] A first aspect of the invention provides a NOX4 inhibitor or a dual NOX4 / 1 inhibitor or a NOX1 inhibitor for use in the treatment of solid tumor cancers exhibiting or susceptible to exhibiting resistance to immunotherapy or an antiangiogenic agent, in particular to antiVEGF treatment, wherein said NOX4 inhibitor (or NOX4 / 1 or NOX1) must be administered in combination with an anticancer immunotherapeutic agent or an antiangiogenic agent.

[0020] Another aspect of the invention provides for the use of one or more NOX4 or NOX4 / 1 dual or NOX1 inhibitors for the preparation of a pharmaceutical composition for the treatment of solid tumor cancers exhibiting or likely to exhibit resistance to immunotherapy or an antiangiogenic agent, in particular to antiVEGF treatment, wherein said one or more NOX4 or NOX4 / 1 or NOX1 inhibitors must be administered in combination with an anticancer immunotherapeutic agent or an antiangiogenic agent.

[0021] Another aspect of the invention relates to a composition Petition 870250096065, dated 10 / 21 / 2025, page 22 / 90 / 51 pharmaceutical containing at least one NOX4 or NOX4 / 1 or NOX1 inhibitor according to the invention, as well as tautomers, geometric isomers, optically active forms and pharmaceutically acceptable salts thereof combined with at least one anticancer immunotherapeutic agent or at least one additional antiangiogenic agent and at least one pharmaceutically acceptable carrier, diluent or excipient thereof.

[0022] Another aspect of the invention relates to a method for treating an individual suffering from a solid tumor cancer exhibiting or susceptible to exhibiting resistance to immunotherapy or an antiangiogenic agent, in particular to an antiVEGF treatment, said method comprising administering an effective amount of one or more NOX4 or NOX4 / 1 or NOX1 inhibitors, in combination with an anticancer immunotherapeutic agent or an antiangiogenic agent in an individual in need thereof.

[0023] Another aspect of the invention relates to a method for restoring or increasing responsiveness to anticancer immunotherapy, in particular restoring sensitivity to immunotherapeutic treatment, notably changing cold tumors to a changing state, in an individual, said method comprising administering an effective amount of one or more NOX4 or NOX4 / 1 or NOX1 inhibitors or a pharmaceutical formulation thereof in combination with an anticancer immunotherapeutic agent in an individual in need thereof.

[0024] Another aspect of the invention relates to a method for restoring or increasing responsiveness to anticancer antiangiogenesis, in particular restoring sensitivity to antiVEGF treatment or preventing resistance to antiVEGF treatment in an individual, said method comprising administering an effective amount of one or more NOX4 or NOX4 / 1 or NOX1 inhibitors or a pharmaceutical formulation of Petition 870250096065, dated 10 / 21 / 2025, page 23 / 90 / 51, the same in combination with an antiangiogenic agent in an individual in need of the same.

[0025] Other features and advantages of the invention will become apparent from the following detailed description. Description of the figures

[0026] Figure 1 shows the effects of treatment with a NOX4 inhibitor (GKT) on the relocation of CD8+ T cells within 4T1 tumors when cancer cells were co-injected with cancer-associated fibroblasts (CAFs) orthotopically within the breast fat pad as described in Example 1. A: Increase in tumor volume expressed in mm3 versus days after injection (arrow) of the combination of tumor cells with CAFs and vehicle (1) or the combination of tumor cells with CAFs and NOX4 inhibitor (2); B: Immunochemistry and quantification thereof showing the efficacy of treatment with the NOX4 inhibitor in reducing CAF-positive SMA tumors; C: Immunochemistry (and quantification thereof) showing that treatment with the NOX4 inhibitor results in the relocation of CD8+ T cells from the tumor edge to the tumor center.

[0027] Figure 2 shows the effects of a combination of aPD1 with a NOX4 inhibitor (GKT) on the therapeutic response in CAF-rich tumors where MC38 cancer cells were co-injected with cancer-associated fibroblasts (CAF) in mice treated as described in Example 1 and the effects of a vehicle alone (Ctl), aPD1, NOX4 inhibitor (GKT) alone or a combination of aPD1 + NOX4 inhibitor (GKT) are compared in terms of tumor growth after injection (A); B: Immunochemistry and quantification thereof showing that treatment with the aPD1 / NOX4 inhibitor combination results in the relocation of CD8+ T cells from the tumor edge to the tumor center compared to aPD1 alone; C: Kaplan-Meier survival curves Petition 870250096065, dated 10 / 21 / 2025, page 24 / 90 / 51 in the various groups.

[0028] Figure 3 shows the effects of a combination of an antitumor vaccination with a NOX4 inhibitor (GKT) as described in Example 2. A: Tumor growth after injection in mice treated with a combination vaccine / GKT compared with the vaccine alone and controls; B: Immunochemistry and quantification of the same showing that treatment with the vaccine / NOX4 inhibitor combination results in the relocation of CD8+ T cells from the tumor edge to the tumor center compared with the vaccine alone; C: Kaplan-Meier survival curves in the various groups.

[0029] Figure 4 shows the efficacy of the combination of an antiangiogenic agent and a selective NOX1 inhibitor (GKT2) in inhibiting angiogenesis as measured by CD45- / CD31+ / GP38- cells as described in Example 3 when compared to controls (*p<0.05; **p<0.01; ***p<0.005; ****p<0.001).

[0030] Figure 5 shows tumor size growth in NOX1-KO mice when compared to WT mice and the effect of an anti-VEGFR2 antibody (DC101) on decreasing tumor growth in these mice. Detailed Description of the Invention

[0031] The term “NOX inhibitor” as used herein refers to any substance capable of totally or partially inhibiting, blocking, attenuating, or interfering with NOX4 and / or NOX1. The term is directly defined as the compound affecting the enzyme's enzymatic activity, cellular localization, protein stability, messenger RNA, or protein expression. Preferably, a NOX4 / NOX1 inhibitor should be able to decrease enzyme activity and ROS production in a cell-free assay using a membrane expressing only the NOX isoform of the NOX4 / 1 protein, such as recombinant NOX4 / 1 protein. Thus, Petition 870250096065, dated 10 / 21 / 2025, page 25 / 90 / 51, the term “inhibitors” is intended to include, but is not limited to, molecules that completely or partially inhibit the activity of NADPH oxidase 4 and / or NADPH oxidase 1. According to a particular embodiment, NOX4 / 1 inhibitors have a higher NOXy inhibitor activity component for NOX4 and / or NOX1 compared to other NOX proteins, for example, for NOX2 and / or NOX3 / 5. According to a particular embodiment, NOX4 / 1 inhibitors have a higher NOXy inhibitor activity at NOX4 / 1 that is at least five times higher than at other NOX proteins.

[0032] For example, NOX4 / 1 inhibitors include small molecules, peptides, peptidomimetics, chimeric proteins, natural or non-natural proteins, nucleic acid-derived polymers (such as DNA and RNA aptamers, siRNAs, shRNAs, PNAs or LNAs), fusion proteins with NOX4 / 1 antagonistic activities, antibody antagonists such as anti-NOX4 / 1 neutralizing antibodies or gene therapy vectors directing the expression of such NOX4 / 1 antagonists.

[0033] In particular, NOX4 / 1 inhibitors are agents that exhibit a Ki inhibitory constant of less than 5 micromolars in a functional ROS production assay such as that described in Gaggini et al., 2011, Bioorganic and Medicinal Chemistry, Vol. 19(23), 69896999. For example, NOX4 / 1 inhibitors are agents that inhibit ROS production in a range of about less than 1 microM, such as between about 30 and 300 nanomolars in a cell-free assay using membrane expression only of the NOX isoform, NOX4 protein or NOX1, such as recombinant NOX4 or NOX1 protein.

[0034] The term “siRNA” refers to small interfering RNA, which is double-stranded RNA (approximately 19 to 23 nucleotides) capable of knocking down or silencing a targeted mRNA of a target gene. Artificial siRNAs can be chemically synthesized as oligonucleotides or Petition 870250096065, of 10 / 21 / 2025, page 26 / 90 / 51 cloned within a plasmid or a viral vector (adenovirus, retrovirus or lentivirus) as short hairpin RNAs to generate a transient or stable transfection in any type of cells (Martin et al., 2007, Ann. Rev. Genomics Hum. Genet., 8: 81-108; Huang et al., 2008, Expert. Opin. Ther. Targets, 12(5), 637-645).

[0035] The term “solid tumor cancer” includes glioblastoma, lung cancer (small cell and non-small cell), breast cancer, ovarian cancer, cervical cancer, uterine cancer, head and neck cancer, melanoma, hepatocellular carcinoma, colon cancer, rectal cancer, colorectal carcinoma, kidney cancer, prostate cancer, gastric cancer, bronchial cancer, pancreatic cancer, bladder cancer, liver cancer and brain cancer, in particular glioblastoma.

[0036] As used herein, “treatment” and “treating” and the like generally mean to obtain a desired pharmacological and physiological effect. The term “treatment” as used herein encompasses any treatment of a disease in a mammal, particularly a human being, and includes inhibiting the disease, i.e., stopping its development, or alleviating the disease, i.e., causing the regression of the disease and / or its symptoms or conditions, such as stopping tumor growth or tumor regression.

[0037] The term “individual” as used herein refers to mammals. For example, mammals considered by the present invention include humans, primates, domesticated animals such as cattle, sheep, pigs, horses, laboratory rodents, dogs and the like.

[0038] The term “effective quantity” as used herein refers to an amount of at least one particle or a pharmaceutical formulation thereof according to the invention that evokes the biological or medicinal response in a tissue, system, animal or human being that is being sought. In one embodiment, the effective quantity is a “therapeutically effective quantity” for the relief of the symptoms of the disease or condition. Petition 870250096065, dated 10 / 21 / 2025, page 27 / 90 / 51 being treated. Typically, an effective amount can be used to inhibit the growth of cancer cells, that is, any decrease in the rate of proliferation and / or migration of cancer cells, cessation of proliferation and / or migration of cancer cells, or extermination of cancer cells, such that the growth rate of cancer cells is reduced compared to the observed or predicted growth rate of an untreated control cancer cell. The term "inhibits growth" can also refer to a reduction in the size or disappearance of a cancer cell or tumor, as well as a reduction in its metastatic potential. Preferably, such inhibition at the cellular level can reduce the size, halt the growth, reduce the aggressiveness, or prevent or inhibit the metastasis of a cancer in a patient.Those skilled in the art can easily determine, by any one of a variety of suitable distinctive markers, whether cancer cell growth is inhibited.

[0039] The term “efficacy” of a treatment according to the invention can be measured based on changes in the course of a disease in response to a use or method according to the invention. The efficacy of a cancer treatment according to the invention can be measured by a reduction in tumor volume and / or an increase in progression-free survival time and / or increased health and well-being of the individual (e.g., suppression of a cancer). Inhibition of cancer cell growth can be evidenced, for example, by the arrest of cancer cells at a particular phase of the cell cycle, for example, arrest at the G2 / M phase of the cell cycle. Inhibition of cancer cell growth can also be evidenced using well-known imaging methods such as magnetic resonance imaging, computed tomography, PET, SPECT, photoacoustic imaging, X-rays, and fluorescence imaging / detection.Cancer cell growth can also be determined indirectly, for example by... Petition 870250096065, dated 10 / 21 / 2025, page 28 / 90 / 51 determination of circulating carcinoembryonic antigen levels, prostate-specific antigen, or other cancer-specific antigens that are correlated with cancer cell growth.

[0040] In particular, the effectiveness of a combined treatment according to the invention can be evaluated by the reduction in tumor size or the disappearance of the tumor or any biomarker relevant to a type of cancer.

[0041] Unless otherwise required by the definition of the individual substituent, the term “substituted” refers to groups substituted with 1 to 5 substituents selected from the group consisting of “C1-C6 alkyl”, “C2-C6 alkenyl”, “C2-C6 alkynyl”, “C3-C8 cycloalkyl”, “heterocycloalkyl”, “C1-C6 aryl alkyl”, “C1-C6 heteroaryl alkyl”, “C1-C6 cycloalkyl alkyl”, “C1-C6 heterocycloalkyl”, “amino”, “alkyl amino”, “aminosulfonyl”, “ammonium”, “alkoxy”, “acyl”, “acyl amino”, “amino carbonyl”, “aryl”, “heteroaryl”, “sulfinyl”, “sulfonyl”, “sulfonamide”, “alkoxy”, “alkoxy carbonyl”, “carbamate”, “sulfanyl”, “halogen”, trihalomethyl, cyano, hydroxy, Mercapto, nitro and similar substances.

[0042] The term “pharmaceutically acceptable salts or complexes” refers to salts or complexes of the compounds specified below of the invention. Examples of such salts include, but are not limited to, base addition salts formed by the reaction of compounds of the invention with organic or inorganic bases such as hydroxide, carbonate, bicarbonate or the like, of a metallic cation such as those selected from the group consisting of alkali metals (sodium, potassium or lithium), alkaline earth metals (e.g., calcium or magnesium) or with a primary, secondary or tertiary organic alkylamine. Other examples of such salts include, but are not limited to, acid addition salts formed by the reaction of compounds of the invention with organic or inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, para-toluenesulfonic acid, 2-naphthalenesulfonic acid, Petition 870250096065, dated 10 / 21 / 2025, page 29 / 90 / 51 camphorsulfonic acid, benzenesulfonic acid, oxalic acid or similar.

[0043] “Pharmaceutically active derivative” refers to any compound that, upon administration to the recipient, is capable of providing, directly or indirectly, the activity disclosed herein. NOX4 / NOX1 inhibitors according to the invention

[0044] In one embodiment, the invention provides a NOX4 or NOX4 / 1 inhibitor or a NOX1 inhibitor having an inhibitory constant (Ki) for NOX4 and / or NOX1 ranging from 60 nM or lower to 300 nM in a ROS production functional assay and wherein the inhibitory activity against other selected NOXs of NOX2, 3 and 5 is higher than 1 micromolar.

[0045] According to a particular embodiment, the NOX4 or NOX4 / NOX1 or NOX1 inhibitors according to the invention are pyrazolopyridine compounds, pyrazolinodione compounds or thiazole starch compounds, as described in WO 2008 / 113856, WO 10 / 035217, WO 10 / 035219, WO 10 / 035220, WO 10 / 035221, WO 11 / 036651, WO 2013 / 068972, WO 2015 / 049655 and WO 2016 / 098005.

[0046] According to another particular embodiment, the NOX4 inhibitors according to the invention are 2,5-disubstituted benzoxazole and benzothiazole derivatives as described in WO 2016 / 207785.

[0047] In one embodiment, the invention provides a NOX4 inhibitor of Formula (I) (I) wherein Gi is selected from H, optionally substituted alkyl such as aminocarbonyl alkyl (e.g., phenylacetamide), optionally substituted C3-C8 cycloalkyl alkyl, heterocycloalkyl alkyl Petition 870250096065, of 21 / 10 / 2025, p. 30 / 90 / 51 optionally substituted, optionally substituted aryl alkyl such as optionally substituted phenyl alkyl such as optionally substituted phenyl methyl (for example, phenyl methyl or 3-methyl phenyl methyl or 4-fluorobenzyl or 2-chlorobenzyl or 4-chlorobenzyl or 4-methyl benzyl or 4-bromobenzyl) and optionally substituted heteroaryl alkyl such as optionally substituted pyridino alkyl such as pyridino-2-yl methyl; G2 is selected from H; optionally substituted alkyl; optionally substituted alkenyl; optionally substituted alkynyl; optionally substituted aryl groups such as optionally substituted phenyl groups (for example, phenyl or 4-fluorophenyl or 4-methoxyphenyl or 4-nitrophenyl or 2-chlorophenyl or 2-methylphenyl or 4-(trifluoromethyl)phenyl or 4-(trifluoromethoxy)phenyl or 2,5-difluorophenyl or 2-methoxyphenyl); optionally substituted alkyl aryl groups; optionally substituted aryl alkyl groups;optionally substituted heteroaryl, such as optionally substituted benzothiazolyl (for example, 1,3-benzothiazol-2-yl) or optionally substituted pyridinyl (for example, pyridin-2-yl); optionally substituted alkyl heteroaryl; optionally substituted heteroaryl alkyl; optionally substituted alkenyl aryl; optionally substituted aryl alkenyl; optionally substituted alkenyl heteroaryl; optionally substituted heteroaryl alkenyl; optionally substituted C3-C8 cycloalkyl; optionally substituted heterocycloalkyl; optionally substituted C3-C8 alkyl cycloalkyl; optionally substituted C3-C8 cycloalkyl alkyl; optionally substituted alkyl heterocycloalkyl and optionally substituted heterocycloalkyl alkyl; G3 is selected from H; optionally substituted alkyl such as methyl or ethyl; optionally substituted alkenyl; optionally substituted alkynyl;optionally substituted aryl such as optionally substituted phenyl (e.g., phenyl); optionally substituted alkyl aryl; optionally substituted aryl alkyl; optionally substituted heteroaryl; optionally substituted alkyl heteroaryl; optionally substituted heteroaryl alkyl; Petition 870250096065, of 10 / 21 / 2025, page 31 / 90 / 51 substituted; optionally substituted alkenyl aryl; optionally substituted aryl alkenyl; optionally substituted alkenyl heteroaryl; optionally substituted heteroaryl alkenyl; optionally substituted C3-C8 cycloalkyl; optionally substituted heterocycloalkyl; optionally substituted C3-C8 alkyl cycloalkyl; optionally substituted C3-C8 cycloalkyl; optionally substituted alkyl heterocycloalkyl and optionally substituted heterocycloalkyl alkyl; G4 is selected from H, optionally substituted alkyl such as optionally substituted pentyl (e.g., isopentyl) or optionally substituted heteroalkyl such as optionally substituted methoxy (e.g., 2-methoxyethyl); optionally substituted alkenyl; optionally substituted alkynyl; optionally substituted aryl; optionally substituted alkyl aryl;optionally substituted aryl alkyl such as optionally substituted phenyl methyl (e.g., methyl or benzyl benzoic acid) or optionally substituted phenyl ethyl (e.g., 2-phenyl ethyl, 4-methoxyphenyl ethyl); optionally substituted heteroaryl; optionally substituted alkyl heteroaryl;optionally substituted heteroaryl alkyl such as optionally substituted thiophenyl alkyl such as optionally substituted thiophenyl methyl (e.g., thiophen-2-yl methyl) or optionally substituted imidazolyl alkyl such as optionally substituted imidazolyl ethyl (e.g., imidazol-4-yl ethyl) or optionally substituted indolyl alkyl such as optionally substituted indolyl ethyl (e.g., indole-3-yl ethyl) or optionally substituted furanyl alkyl such as optionally substituted furanyl methyl (e.g., furan-2-yl methyl) or optionally substituted benzodioxolyl alkyl such as optionally substituted benzodioxolyl methyl (e.g., 1,3-benzodioxol-5-yl methyl) or optionally substituted pyridinyl alkyl such as optionally substituted pyridinyl methyl (e.g., pyridino-3-yl methyl or pyridin-2-yl methyl); optionally substituted alkenyl aryl; aryl alkenyl optionally; Petition 870250096065, dated 10 / 21 / 2025, page 32 / 90 / 51 substituted; optionally substituted alkenyl heteroaryl; optionally substituted heteroaryl alkenyl; optionally substituted C3-C8 cycloalkyl; optionally substituted heterocycloalkyl such as optionally substituted morpholinyl (e.g., 5-morpholinnyl-4-yl) or optionally substituted piperazinyl (e.g., 4-methylpiperazinyl) or optionally substituted piperidinyl (e.g., 4-methylbenzyl)piperidin-4-yl); optionally substituted C3-C8 alkyl cycloalkyl and optionally substituted C3-C8 cycloalkyl;optionally substituted alkyl heterocycloalkyl and optionally substituted heterocycloalkyl alkyl such as optionally substituted morpholinyl alkyl such as optionally substituted morpholinyl propyl (e.g., 3-(morpholin-4-yl)propyl) optionally substituted morpholinyl ethyl (e.g., 2-morpholin-4-ylethyl) or optionally substituted piperazinyl alkyl such as optionally substituted piperazinyl ethyl (e.g., 2-(4-acetylpiperazin-1-yl)ethyl or 2-(4-hexanoylpiperazin-1-yl)ethyl) or optionally substituted pyrrolidinyl alkyl such as optionally substituted pyrrolidinyl propyl (e.g., 3-(2-oxopyrrolidin-1-yl)propyl) or optionally substituted tetrahydrofuranyl alkyl such as optionally substituted tetrahydrofuranyl methyl (e.g., tetrahydrofuran-2-yl methyl); G5 is selected from H, optionally substituted alkyl; optionally substituted alkenyl; optionally substituted alkynyl;optionally substituted aryl; optionally substituted alkyl aryl; optionally substituted aryl alkyl; optionally substituted heteroaryl; optionally substituted alkyl heteroaryl; optionally substituted heteroaryl alkyl; optionally substituted alkenyl aryl; optionally substituted aryl alkenyl; optionally substituted heteroaryl alkenyl; optionally substituted heteroaryl alkenyl; optionally substituted C3-C8 cycloalkyl; optionally substituted heterocycloalkyl; optionally substituted C3-C8 alkyl cycloalkyl; optionally substituted C3-C8 cycloalkyl alkyl; alkyl; Petition 870250096065, dated 10 / 21 / 2025, page 33 / 90 / 51 optionally substituted heterocycloalkyl and optionally substituted heterocycloalkyl alkyl; as well as pharmaceutically acceptable salts and pharmaceutically active derivatives thereof.

[0048] In another embodiment, the invention provides a NOX4 / 1 inhibitor Formula (II) wherein Ar is optionally substituted phenyl such as halogen-substituted phenyl such as chlorine (e.g., 2-chlorophenyl) or alkoxy-substituted phenyl (e.g., methoxy); Gi and G4 are H; G2 is selected from optionally substituted C1-C6 alkyl (e.g., methyl) and optionally substituted phenyl (such as halogen-substituted phenyl such as 3-chlorophenyl, 4-chlorophenyl, 2-fluorophenyl, 3-fluorophenyl, 4-fluorophenyl, 4-chloro-2-fluorophenyl, 5-chloro-2-fluorophenyl, amino or alkyl amino-substituted phenyl or alkoxy-substituted phenyl such as 3-dimethylaminophenyl, 2-trimethylaminophenyl, 3-methylaminophenyl, 3-aminophenyl, 4-methoxyphenyl);G3 is selected from H, optionally substituted C1-C0 alkyl (e.g., methyl, alkoxy-substituted C1-C6 alkyl such as methoxyethyl or 2-methoxyethyl), optionally substituted C1-C0 heteroaryl alkyl such as optionally substituted C1-C0 pyridinyl alkyl (e.g., optionally substituted pyridinyl methyl such as 2-pyridinyl-2-ylmethyl, 3-pyridinyl-3-ylmethyl, o-methoxypyridin-3-ylmethyl, 2-methoxypyridin-4-ylmethyl) or optionally substituted C1-C0 pyrazinyl alkyl (e.g., 2-pyrazinyl-2-ylmethyl) and optionally substituted C1-C0 alkoxy alkyl such as methoxyethyl (e.g., 2-methoxyethyl) or G2 and G3 together form an optionally substituted 7-membered heterocycloalkyl ring comprising; Petition 870250096065, dated 10 / 21 / 2025, p. 34 / 90 / 51 two nitrogen atoms and where the two nitrogens are joined through an optionally substituted C1-C3 alkyl moiety, as well as tautomers, geometric isomers, optically active forms and pharmaceutically acceptable salts thereof.

[0049] In a particular embodiment, the invention provides a NOX4 / 1 inhibitor of Formula (II) wherein G2 and G3 together form an optionally substituted 7-membered heterocycloalkyl ring comprising two nitrogen atoms to form the following compound of Formula (I'): where Ar, G1 and G5 are as defined here; G6, G8 to G10 are H; G7 is selected from optionally substituted C1-C6 alkyl such as optionally substituted C1-C6 alkyl with optionally substituted phenyl (e.g., optionally substituted methyl with optionally substituted phenyl such as benzyl, optionally substituted methyl with halogen-substituted phenyl such as 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, optionally substituted methyl with alkoxy-substituted phenyl such as 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl), optionally substituted C1-C6 aryl alkyl such as optionally substituted C1-C6 phenyl alkyl (e.g., benzyl, 2-chlorobenzyl, 3-chlorobenzyl, 4-chlorobenzyl, 2-methoxybenzyl, 3-methoxybenzyl, 4-methoxybenzyl) and optionally substituted C1-C6 heteroaryl alkyl such as pyridinyl alkyl. Optionally substituted C1-Cô (e.g., optionally substituted pyridinyl methyl such as pyridinyl-2-ylmethyl,pyridinyl-3-ylmethyl) or optionally substituted C1-C6 furanyl alkyl (e.g., optionally substituted methyl furanyl such as furan-3-ylmethyl) thus, Petition 870250096065, dated 10 / 21 / 2025, p. 35 / 90 / 51 as tautomers, geometric isomers, optically active forms and pharmaceutically acceptable salts thereof.

[0050] In a particular embodiment, the invention provides a compound of Formula (II) for use in accordance with the invention wherein G2 is optionally substituted C1-C6 alkyl.

[0051] In another particular embodiment, the invention provides a compound of Formula (II) for use in accordance with the invention wherein G2 is optionally substituted phenyl.

[0052] In another particular embodiment, the invention provides a compound of Formula (II) for use in accordance with the invention wherein G3 is optionally substituted C1-C6 alkyl.

[0053] In another particular embodiment, the invention provides a compound of Formula (II) for use in accordance with the invention wherein G3 is optionally substituted C1-C6 heteroaryl alkyl such as optionally substituted C1-C6 pyridinyl alkyl.

[0054] In another particular embodiment, the invention provides a compound of Formula (II) for use in accordance with the invention wherein G2 and G3 together form an optionally substituted 7-membered heterocycloalkyl ring comprising two nitrogen atoms to form the following compound of Formula (I'), wherein G7 is an optionally substituted C1-C6 alkyl.

[0055] In another embodiment, the invention provides a compound of Formula (II) for use in accordance with the invention wherein G2 and G3 together form an optionally substituted 7-membered heterocycloalkyl ring comprising two nitrogen atoms to form the following compound of Formula (I'), wherein G7 is an optionally substituted C1-C6 aryl alkyl.

[0056] In another embodiment, the invention provides a compound of Formula (I) for use according to the invention wherein G2 and G3 together form an optionally substituted 7-membered heterocycloalkyl ring comprising Petition 870250096065, dated 10 / 21 / 2025, p. 36 / 90 21 / 51 two nitrogen atoms to form the following compound of Formula (Γ), where G? is optionally substituted C1-C1 heteroaryl alkyl.

[0057] According to another particular embodiment, NOX1 inhibitors according to the invention are thiazole starch derivatives as described in WO 2016 / 098005.

[0058] In another embodiment, a NOX1 inhibitor of Formula (III) is provided: (III) wherein X is selected from CR1 and N; Y is selected from CH or N; Ai is selected from -OCHR5-, -NR4-CHR5-, -CH2NR4- and -CH2-O-; R1 is selected from H, halogen and optionally substituted C1-C1 alkyl; R2 is selected from H, halogen (e.g., chlorine, fluorine), optionally substituted alkoxy such as optionally substituted methoxy (e.g., methoxy, (tetrahydro-2H-pyran-4-yl)methoxy, piperidin-4-ylmethoxy) or optionally substituted ethoxy (e.g., 2-(dimethylamino)ethoxy, 2-hydroxyethoxy, 1-phenylethoxy, 2-methoxyethoxy), optionally substituted C1-C1 alkyl alkoxy, optionally substituted C1-C1 alkyl such as optionally substituted methyl, optionally substituted amino such as optionally substituted C1-C1 alkyl amino (e.g., methyl amino, tetrahydro-2H-pyran-4-yl)methyl)amino, (1-methylpiperidin-4-yl)methyl)amino, dimethyl amino, optionally substituted ethyl amino such as 2-morpholino ethyl amino or 2-(dimethylamino) ethyl amino or methoxy ethyl amino,optionally substituted methyl amino such as 1-methyl-1H-imidazol-4-yl methyl amino or 2-hydroxyethyl)amino, optionally substituted propyl amino such as dimethylamino propyl amino), optionally substituted heterocycloalkyl such as optionally substituted piperazine (for example, methylpiperazin-1, Petition 870250096065, dated 10 / 21 / 2025, p. 37 / 90 / 51 ila), optionally substituted C1-C1 alkyl heterocycloalkyl such as optionally substituted C1-C1 alkyl piperazine (e.g., methylpiperazin-l-ila), optionally substituted C1-C1 amino alkyl, optionally substituted C1-C1 alkoxy alkyl, -O-R8 and -NR9R10; R3 is a group of Formula -(CHR6)n-A2 or R3 forms with the CHR5 portion of Ai an optionally substituted ring selected from optionally substituted aryl such as an optionally substituted phenyl (e.g., phenyl or halogen-substituted phenyl such as fluorine, alkoxy-substituted phenyl such as methoxy) and optionally substituted heteroaryl such as optionally substituted 1,3-dihydro-1H-indenyl (e.g., 1-(dimethylamino)-2,3-dihydro-1H-inden-2-yl, 2,3-dihydro-1H-inden-2-yl, 2,3-dihydro-1H-inden-1yl) or optionally substituted 6,7-dihydro-5H-cyclopentapyridinyl (e.g., 6,7-dihydro-5H-cyclopenta[b]pyridin-5-yl,2-methylpyridin-3-yl, 5-methylpyridin-2-yl) or optionally substituted 1,2,3,4-tetrahydronaphthalenyl (e.g., 1,2,3,4-tetrahydronaphthalen-1-yl) or optionally substituted 2,3-dihydrobenzofuranyl (e.g., 2,3-dihydrobenzofuran-3-yl, 2,3-dihydro-1H-inden-1-yl) or optionally substituted thiadiazolyl (e.g., 1,3,4-thiadiazol-2-yl) or optionally substituted isoxazolyl (e.g., 5-methylisoxazol-3-yl) or optionally substituted pyrazolyl (e.g., 1-methyl-1H-pyrazol-3-yl) or optionally substituted imidazolyl (e.g., 1-methyl-1H-imidazol-2-yl) or R3form with the NR4 moiety A1 is an optionally substituted ring selected from optionally substituted aryl and optionally substituted heteroaryl such as optionally substituted isoindolinyl (e.g., isoindolin-2-yl, 1H-indol-1-yl); n is an integer from 0 to 4 (such as 0, 1, 2,3 or 4); R4 is selected from H and optionally substituted alkyl such as optionally substituted methyl; A2 is an optionally substituted ring selected from optionally substituted aryl such as optionally substituted phenyl (e.g., methoxyphenyl, fluorophenyl, chlorophenyl), optionally substituted heteroaryl such as, Petition 870250096065, dated 10 / 21 / 2025, page. 38 / 90 / 51 as optionally substituted pyridine (e.g., pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, 2-methylpyridin-3-yl, 5-methylpyridin-2-yl) or optionally substituted pyrazolyl (e.g., 1,3-dimethyl-1H-pyrazol-5-yl, 1-methyl-1H-pyrazol-3-yl) or optionally substituted thiadiazolyl (e.g., 1,3,4-thiadiazol-2-yl) or optionally substituted imidazolyl (e.g., 1H-imidazol-4-yl, 1-methyl-1H-imidazol-2-yl, 1-methyl-1H-imidazol-5-yl) or optionally substituted 1,2,4-triazolyl (e.g., 1-methyl-1H-1,2,4-triazol-5-yl) or optionally substituted isoxazolyl (e.g., 1-cyclopropylisoxazol-3-yl) or optionally substituted oxadiazolyl (e.g., 5-methyl-1,2,4-oxadiazol-3-yl) or optionally substituted pyrimidinyl (e.g., pyrimidinyl-2-yl); R5 is selected from optionally substituted C1-C1 alkyl such as optionally substituted methyl (e.g., methoxymethyl, 3-C1 ...3-difluoropyrrolidin-1-yl methyl, 4-methylpiperazin-1-yl methyl, hydroxyl methyl) or optionally substituted ethyl or optionally substituted propyl (e.g., methyl, hydroxymethyl, hydroxyethyl, 2-propanolyl, hydroxyl isopropyl), optionally substituted C1-C6 amino alkyl such as optionally substituted amino methyl (e.g., dimethylamino methyl, methylamino methyl), optionally substituted C1-C6 alkoxy alkyl, optionally substituted C1-C6 heterocycloalkyl alkyl such as optionally substituted heterocycloalkyl methyl, e.g., optionally substituted C1-C6 pyrrolidin alkyl (e.g., 3,3-difluoropyrrolidin-1-yl methyl) or substituted C1-C6 piperazine alkyl (e.g., 4-methylpiperazin-1-yl methyl) or ethyl heterocycloalkyl, e.g., C1-C6 morpholino alkyl optionally substituted (e.g., methyl morpholino, ethyl morpholino) or optionally substituted C1-C6 alkyl pyrrolidin (e.g., methyl pyrrolidin, ethyl pyrrolidin),optionally substituted aminocarbonyl (e.g., dimethyl aminocarbonyl), optionally substituted C2-C8 cycloalkyl such as optionally substituted cyclopropyl, and optionally substituted C1-C6 aminoalkyl such as amino, Petition 870250096065, dated 10 / 21 / 2025, page 39 / 90 / 51 optionally substituted ethyl (e.g., dimethyl amino ethyl) or optionally substituted amino methyl (e.g., dimethyl amino methyl); R6 is selected from H, optionally substituted C1-C6 alkyl such as optionally substituted methyl, optionally substituted C1-C6 amino alkyl, optionally substituted amino (e.g., dimethyl amino) and hydroxy and wherein R6 groups are independently selected for each repeating unit (CHR6); R7 is selected from H, halogen (e.g., fluorine) and optionally substituted C1-C0 alkyl such as methyl;R8 is selected from H, optionally substituted C1-C6 alkyl such as optionally substituted methyl or optionally substituted ethyl (e.g., methoxyethyl, 2-(dimethylamino)ethyl, hydroxyethyl), optionally substituted C1-C0 amino alkyl, optionally substituted heterocycloalkyl, optionally substituted C2-C8 cycloalkyl, optionally substituted C1-C0 heterocycloalkyl alkyl such as optionally substituted methyl heterocycloalkyl, e.g., optionally substituted tetrahydropyran C1-C0 alkyl (e.g., tetrahydro-2H-pyran-4-yl) or optionally substituted piperidin alkyl (e.g., 1-methylpiperidin-4-yl), optionally substituted C2-C8 cycloalkyl C1-C0 alkyl, optionally substituted alkoxy, optionally substituted C1-C0 amino alkyl such as optionally substituted amino ethyl (e.g., 2-(dimethylamino)ethyl); optionally substituted aryl C1-C6 alkyl and optionally substituted heteroaryl C1-C0 alkyl;R9 and R10 are independently selected from H, optionally substituted C1-C6 alkyl such as an optionally substituted methyl (e.g., 1-methyl-1H-imidazol-4-yl)methyl) or optionally substituted ethyl (e.g., 2-methoxyethyl), optionally substituted C1-C0 aminoalkyl such as optionally substituted aminoethyl (e.g., dimethylaminoethyl) or optionally substituted aminopropyl (e.g., dimethylamino)propyl), optionally substituted heterocycloalkyl such as optionally substituted piperidine (e.g., 1; Petition 870250096065, dated 10 / 21 / 2025, page. 40 / 90 / 51 methylpiperidine), optionally substituted C2-C8 cycloalkyl, optionally substituted C1-C6 heterocycloalkyl alkyl such as optionally substituted ethyl heterocycloalkyl, for example optionally substituted C1-C6 morpholino alkyl (e.g., 2-morpholino ethyl) or optionally substituted methyl heterocycloalkyl, for example optionally substituted C1-C6 tetrahydrofuran alkyl (e.g., tetrahydro-2H-pyran-4-yl methyl) or optionally substituted C1-C6 piperidin alkyl (e.g., 1-methylpiperidin-4-yl) methyl or optionally substituted C1-C6 imidazolyl alkyl (e.g., 1-methyl-1H-imidazol-4-yl) methyl), optionally substituted C2-C8 cycloalkyl, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 alkoxy alkyl such as optionally substituted ethyl alkoxy (for example, 2-methoxyethyl),Optionally substituted C1-C6 aryl alkyl and optionally substituted C1-C6 heteroaryl alkyl such as methyl C1-C6 heteroaryl alkyl, for example optionally substituted C1-C6 imidazolyl alkyl (e.g., 1-methyl-1H-imidazol-4-yl methyl), optionally substituted C1-C6 amino alkyl such as optionally substituted amino ethyl or optionally substituted amino propyl (e.g., 2-(dimethylamino)ethyl, 2-(dimethylamino)propyl); as well as tautomers, geometric isomers, optically active forms, pharmaceutically acceptable salts and pharmaceutically active derivatives thereof.

[0059] In a particular embodiment, the invention provides a compound of Formula (III) for use in accordance with the invention wherein X is CH.

[0060] In a particular embodiment, the invention provides a compound of Formula (III) for use in accordance with the invention wherein Y is CR1, in particular CH.

[0061] In a particular embodiment, the invention provides a compound of Formula (III) for use in accordance with the invention wherein R2 is Petition 870250096065, dated 10 / 21 / 2025, p. 41 / 90 26 / 51 optionally substituted alkoxy (e.g., methoxy).

[0062] In a particular embodiment, the invention provides a compound of Formula (III) for use in accordance with the invention wherein R7 is H.

[0063] In a particular embodiment, the invention provides a compound of Formula (III) for use in accordance with the invention wherein Ai is -OCHR5, in particular wherein R5 is an optionally substituted C1-C1 morpholino alkyl (e.g., morpholino methyl).

[0064] In another particular embodiment, the invention provides a compound of Formula (III) for use in accordance with the invention wherein Ai is -OCHR5, in particular wherein R5 is an optionally substituted C1-C1 amino alkyl (e.g., dimethyl amino methyl).

[0065] In another particular embodiment, the invention provides a compound of Formula (III) for use in accordance with the invention wherein Ai is -OCHR5, in particular wherein R5 is an optionally substituted C1-C1 hydroxyl alkyl (e.g., hydroxymethyl).

[0066] In a particular embodiment, the invention provides a compound of Formula (III) for use in accordance with the invention wherein R3 is a group of Formula -(CHR6)n-A2, in particular wherein n is 0 and A2 is optionally substituted phenyl (e.g., phenyl).

[0067] According to another particular embodiment, a NOX1 inhibitor according to the invention is 3-methoxy-4-(2-morpholino-1-phenylethoxy)N-(5-(pyridin-4-yl)-1,3,4-thiadiazol-2-yl)benzamide, in particular (R) 3-methoxy4-(2-morpholino-1 -phenylethoxy)-N-(5-(pyridin-4-yl)-1,3,4-thiadiazol-2-yl)benzamide.

[0068] In another embodiment, a NOX4 inhibitor of Formula (IV) is provided: Petition 870250096065, of 10 / 21 / 2025, page 42 / 90 / 51 (IV) wherein ring (A) represents a non-aromatic 5- to 7-membered heterocyclic ring that is fused to the phenyl group; wherein said 5- to 7-membered heterocyclic ring contains a ring oxygen atom and optionally an additional ring heteroatom independently selected from oxygen or nitrogen; wherein said 5- to 7-membered heterocyclic ring is independently unsubstituted or mono- or disubstituted, wherein the substituents are independently selected from: • an oxo substituent bonded to a ring carbon atom in the alpha position to a ring oxygen and / or a ring nitrogen atom and 1 or • a C1-3 alkyl group bonded to a ring nitrogen atom having a free valence; or • two fluorine substituents bonded to the same ring carbon atom; L represents -NH-CO-* or -CO-NH-*, where the asterisks (*) indicate the linkage that is attached to the benzoxazole 1 portion of the benzothiazole; X represents O or S and Y represents -NR1R2 where R1 represents C1-4 alkyl; C2-4 alkyl that is monosubstituted with C1-3 di-(C1-3 alkyl), hydroxy or alkoxy; C3-5-L1 cycloalkyl, wherein L1 represents a direct linkage or C1-3 alkylene and wherein the C3-5 cycloalkyl optionally contains a ring oxygen atom and wherein said C3-5 cycloalkyl is unsubstituted or monosubstituted with methyl or fluorine or a piperidin-3-yl, piperidin-4-yl or pyrrolidin-3-yl group, which groups are substituted at the ring nitrogen atom with C3-5 cycloalkyl, wherein said C3-5 cycloalkyl optionally contains a ring oxygen atom and R2 represents hydrogen, C1-3 alkyl or C3-5 cycloalkyl or Petition 870250096065, dated 10 / 21 / 2025, pp. 43 / 90 / 51 Y represents a saturated 4- to 7-membered monocyclic heterocycline selected from morpholin-4-yl; 2-oxo-pyrrolidin-1-yl; 1,1-dioxythiomorpholin-4-yl or piperazin-1-yl optionally monosubstituted at position 4 with oxetan-3-yl or C1-3 alkyl or azetidin-1-yl, pyrrolidin-1-yl or piperidin-1-yl; wherein said azetidin-1-yl, pyrrolidin-1-yl or piperidin-1-yl is independently unsubstituted or substituted with: • two fluorine substituents attached to the same carbon atom of the ring; or • a substituent selected from unsubstituted phenyl or unsubstituted 6-membered heteroaryl; or • a substituent selected from hydroxy; C1-3 alkoxy; C1-4 -COalkoxy; C1-3 di(alkyl)amino and C1-3 alkyl that is monosubstituted with C1-3 di-(alkyl)amino, hydroxy or alkoxy; or • two substituents, wherein one of said substituents is C1-4 alkyl and the other is independently selected from hydroxy or di-(alkyl C1-3)amino; or • a substituent selected from morpholin-4-yl; 1,1-dioxydothiomorpholin-4-yl or piperazin-1-yl that is optionally monosubstituted at position 4 with C1-3 alkyl; • a substituent selected from azetidin-1-yl, pyrrolidin-1-yl or piperidin-1-yl; wherein said groups are independently unsubstituted or monosubstituted with hydroxyl or disubstituted with methyl and hydroxyl; or Y represents a saturated 7- to 11-membered fused, bridged, or spiro-bicyclic heterocycline containing at least one nitrogen atom, wherein said nitrogen atom is linked to the benzoxazole / benzothiazole moiety and wherein said heterocycline optionally contains an additional ring heteroatom independently selected from Petition 870250096065, dated 10 / 21 / 2025, pp. 44 / 90 / 51 oxygen, nitrogen and sulfur; wherein said heterocycline is unsubstituted or substituted with: - two oxo substituents on a sulfur atom of the ring; or - a C1-3 alkyl substituent attached to a nitrogen atom of the ring having a free valence; or a pharmaceutically acceptable salt thereof.

[0069] In another particular embodiment, a compound of Formula (I) is provided for use in accordance with the invention, wherein the compound is 2-(2-chlorophenyl)-4-methyl-5-(pyridin-2ylmethyl)-1H-pyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione.

[0070] In another particular embodiment, a compound of Formula (I) is provided for use in accordance with the invention, wherein the compound is nh ci 2-(2-chlorophenyl)-4-[3-(dimethylamino)phenyl]5-methyl-1H-pyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione.

[0071] In another particular embodiment, a compound of Formula (I) is provided for use in accordance with the invention, wherein the compound is 4-(2-fluoro-4-methoxyphenyl)-2-(2methoxyphenyl)-5-(pyridin-3-ylmethyl)-1H-pyrazolo[4,3-c]pyridine-3,6(2H,5H)dione.

[0072] In another particular embodiment, a compound is provided Petition 870250096065, dated 10 / 21 / 2025, p. 45 / 90 / 51 of Formula (I') for use in accordance with the invention, wherein the compound is 10-Benzyl-2-(2-chlorophenyl)-2,3,8,9,10,11hexahydro-1H-pyrazolo [4',3':3,4]pyrido [1,2-a][1,4]diazepine-1,5(7H)-dione.

[0073] In another particular embodiment, a compound of Formula (IV) is provided for use in accordance with the invention, wherein the compound (R)-3-methoxy-4-(2-morpholino-1phenylethoxy)-N-(5-(pyridin-4-yl)-1,3,4-thiadiazol-2-yl)benzamide.

[0074] In another particular embodiment, a compound of Formula (IV) is provided for use in accordance with the invention, wherein the compound is: N^\. O y}nh xN' (S)-3-methoxy-4-(1phenylethoxy)-N-(5-(pyridin-4-yl)-1,3,4-thiadiazol-2-yl)benzamide.

[0075] In another particular embodiment, a compound of Formula (IV) is provided for use in accordance with the invention, wherein the compound is: (R)-4-(2-hydroxy-1phenylethoxy)-3-methoxy-N-(5-(pyridin-4-yl)-1,3,4-thiadiazol-2-yl)benzamide.

[0076] In another particular embodiment, a compound of Formula (IV) is provided for use in accordance with the invention, wherein the compound is: Petition 870250096065, dated 10 / 21 / 2025, pp. 46 / 90 / 51 N--( (R)-4-(2-(dimethylamino)-1-phenylethoxy)-3-methoxy-N-(5-(pyridin-4-yl)-1,3,4-thiadiazol-2-yl)benzamide.

[0077] In another particular embodiment, a compound according to the invention is provided selected from the following group: 2-(2-chlorophenyl)-4-methyl-5-(pyridin-2-ylmethyl)-1H-pyrazolo[4,3c]pyridine-3,6(2H,5H)-dione; 2-(2-chlorophenyl)-4-[3-(dimethylamino)phenyl]-5-methyl-1Hpyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione; 4-(2-fluoro-4-methoxyphenyl)-2-(2-methoxyphenyl)-5-(pyridin-3ylmethyl)-1H-pyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione; (R)-3-methoxy-4-(2-morpholino-1-phenylethoxy)-N-(5-(pyridin-4-yl)1,3,4-thiadiazol-2-yl)benzamide; 10-benzyl-2-(2-chlorophenyl)-2,3,8,9,10,11-hexahydro-1Hpyrazolo[4',3':3,4]pyrido[1,2-a][1,4]diazepine-1,5(7H)-dione; (S)-3-methoxy-4-(1-phenylethoxy)-N-(5-(pyridin-4-yl)-1,3,4thiadiazol-2-yl) benzamide; (R)-4-(2-hydroxy-1-phenylethoxy)-3-methoxy-N-(5-(pyridin-4-yl)1,3,4-thiadiazol-2-yl)benzamide and (R)-4-(2-(dimethylamino)-1-phenylethoxy)-3-methoxy-N-(5-(pyridin4-yl)-1,3,4-thiadiazol-2-yl)benzamide.

[0078] According to a particular aspect, a NOX inhibitor selected from a NOX4 inhibitor and a NOX4 / 1 inhibitor is provided for use in combination with a cancer vaccine or with at least one immune checkpoint inhibitor.

[0079] According to a further particular aspect, a NOX inhibitor selected from a NOX4 inhibitor and a NOX4 / 1 inhibitor is provided for use in combination with a cancer vaccine or with Petition 870250096065, dated 10 / 21 / 2025, pp. 47 / 90 / 51, at least one immune checkpoint inhibitor.

[0080] According to another particular aspect, a NOX inhibitor selected from a NOX4 inhibitor and a NOX4 / 1 inhibitor is provided for use in combination with a cancer vaccine.

[0081] According to another particular aspect, a NOX inhibitor selected from a NOX1 inhibitor and a NOX1 / 4 inhibitor is provided for use in combination with at least one antiangiogenic agent. Anticancer immunotherapeutic agents according to the invention

[0082] An anticancer immunotherapeutic agent that can be used according to the invention includes cancer vaccines such as oncolytic vaccines or simple herpesviruses as described in Bartlett et al., 2013, Molecular Cancer 2, 12: 103 (e.g., talimogene laherparepvec (Imlygic)) or in Fukuhara et al., 2016, Cancer Sci, 107(10), 1373-1379, adoptive cellular immunotherapy as described in Perica et al., 2015, Rambam Maimonides Med J, 6(1), e0004, immune checkpoint inhibitors such as PD-1 inhibitors like those described in Iwai et al., 2017, Journal of Biomedical Science, 24:26 or Mishra, 2017, Future Oncol. doi: 10.2217 / fon-2017-0115 or Soto Chervin et al., 2016, F1000Research 2016, 5(F1000 Faculty Rev):803 (for example, such as Pembrolizumab (Keytruda), Nivolumab (Opdivo)) or PD-L1 inhibitors such as Atezolizumab (Tecentriq), Avelumab (Bavencio), Durvalumab (Imfinzi) or CTLA-4 inhibitors such as Ipilimumab (Yervoy).

[0083] According to another particular aspect, an immune checkpoint inhibitor according to the invention may be selected from T-cell immunoglobulin and mucin domains 3 (TIM3), lymphocyte activating gene 3 (LAG3), T-cell immunoglobulin and ITIM domains (TIGIT) or B and T lymphocyte attenuating inhibitors (BTLA).

[0084] According to a particular aspect, an immune checkpoint inhibitor Petition 870250096065, of 10 / 21 / 2025, pp. 48 / 90 / 51 immunological verification according to the invention is a PD-1 inhibitor.

[0085] According to a particular aspect, an anticancer vaccine according to the invention encompasses DNA, RNA, peptide and oncolytic virus vaccines.

[0086] Furthermore, more generally, since the infiltration of CD8+ T cells into tumors is fundamental to most immunotherapies, combinations and combined uses according to the invention would also be useful in adoptive T cell transfer therapies, including tumor-infiltrating lymphocytes (TILs), T cell receptor (TCR) T cells, and chimeric antigen receptor (CAR) T cells as described in June et al., 2018, Science, 359: 1361-1365. TILs have been shown to induce durable, complete responses in patients with metastatic melanoma. CAR T cells have produced significant benefit in the treatment of hematological malignancies (Kochenderfer et al., 2010, Blood 116, 4099-4102; Porter et al., 2011, N. Engl. J. Med., 365, 725-733; Brentjens et al., 2013, Sci. Transl. Med., 5, 177ra38; Grupp et al., 2013, N. Engl. J. Med., 368, 1509-1518), however, the tumor microenvironment remains a significant barrier to success in solid cancers.

[0087] Similarly, the immunotherapeutic agent that can be used according to the invention includes CD8+ T cell agonists, such as αCD40, α-CD27, α-41BB, α-OX40, GITR. Antiangiogenic agents for use in a combination according to the invention.

[0088] An antiangiogenic agent that can be used according to the invention includes antiVEGF agents such as those described in Gardner et al., 2017, supra, in particular bevacizumab or sunitinib. Compositions

[0089] The invention provides pharmaceutical or therapeutic agents as compositions and methods for treating a patient, preferably a patient Petition 870250096065, dated 10 / 21 / 2025, pp. 49 / 90 / 51 mammal and most preferably a human patient suffering from a solid tumor cancer exhibiting or susceptible to exhibiting resistance to immunotherapy or an anti-angiogenic agent, in particular to anti-VEGF treatment.

[0090] The pharmaceutical compositions of the invention may contain one or more compounds in any form described herein. The compositions of this invention may further comprise one or more additional pharmaceutically acceptable ingredient(s), such as alum, solubilizers, stabilizers, antimicrobial agents, buffers, color agents, flavoring agents, adjuvants and the like.

[0091] The compounds of the invention, together with a conventionally used adjuvant, carrier, diluent or excipient, may be placed in the form of pharmaceutical compositions and unit dosages thereof and in such form may be used as solids, such as powder in sachets, tablets or filled capsules, or liquids such as solutions, suspensions, emulsions, elixirs, nasal sprays or capsules filled therewith, all for oral use, or in the form of sterile injectable solutions for parenteral use (including subcutaneous). Such pharmaceutical compositions and unit dosage forms thereof may comprise ingredients in conventional proportions, with or without additional compounds or active ingredients, and such unit dosage forms may contain any effective amount suitable for the active ingredient commensurate with the intended daily oral dosage range to be used.The compositions according to the invention are preferably oral, sublingual, nasal, and subcutaneous.

[0092] The compositions of this invention may also be liquid formulations, including but not limited to aqueous or oily suspensions, solutions, emulsions, syrups, sprays and elixirs. Liquid forms suitable for oral administration may include a suitable aqueous or non-aqueous vehicle with buffers, suspending agents and Petition 870250096065, dated 10 / 21 / 2025, pp. 50 / 90 / 51, dispenses with the use of colorants, flavorings, and similar substances. The compositions may also be formulated as a dry product for reconstitution with water or another suitable vehicle before use. Such liquid preparations may contain additives, including but not limited to suspending agents, emulsifying agents, non-aqueous vehicles, and preservatives. Suspending agents include, but are not limited to, sorbitol syrup, methylcellulose, glucose / sugar syrup, gelatin, hydroxyethylcellulose, carboxymethyl cellulose, aluminum stearate gel, and hydrogenated edible fats. Emulsifying agents include, but are not limited to, lecithin, sorbitan monooleate, and acacia. Non-aqueous vehicles include, but are not limited to, edible oils, almond oil, fractionated coconut oil, oily esters, propylene glycol, and ethyl alcohol.Preservatives include, but are not limited to, methyl or propyl p-hydroxybenzoate and sorbic acid. Additional materials, as well as processing techniques and the like, are presented in The Science and Practice of Pharmacy (Remington: The Science & Practice of Pharmacy), 22nd Edition, 2012, Lloyd, Ed. Allen, Pharmaceutical Press, which is incorporated herein by reference.

[0093] The solid compositions of this invention may be in powder form in sachets, tablets or lozenges formulated in a conventional manner. For example, sachets, tablets and capsules for oral or sublingual administration may contain conventional excipients including, but not limited to, binding agents, fillers, lubricants, disintegrants and wetting agents. Binders include, but are not limited to, syrup, acacia, gelatin, sorbitol, tragacanth, starch mucilage and polyvinylpyrrolidone. Fillers include, but are not limited to, lactose, sugar, microcrystalline cellulose, corn starch, calcium phosphate and sorbitol. Lubricants include, but are not limited to, magnesium stearate, stearic acid, talc, polyethylene glycol and silica. Disintegrants include, but are not limited to, potato starch and sodium starch glycolate. Petition 870250096065, dated 10 / 21 / 2025, page 51 / 90 / 51 Humectants include, but are not limited to, sodium lauryl sulfate. Tablets may be coated according to methods well known in the art.

[0094] Injectable compositions are typically based on sterile injectable saline solution or phosphate-buffered saline solution or other injectable carriers known in the art.

[0095] The compositions of this invention may also be formulated for parenteral administration, including, but not limited to, by injection or continuous infusion. Formulations for injection may be in the form of suspensions, solutions or emulsions in oily or aqueous vehicles and may contain formulation agents including, but not limited to, suspending, stabilizing and dispersing agents. The composition may also be provided in a powder form for reconstitution with a suitable vehicle including, but not limited to, sterile, pyrogen-free water.

[0096] The compositions of this invention can also be formulated as a depot preparation, which can be administered by implantation or intramuscular injection. The compositions can be formulated with suitable polymeric or hydrophobic materials (such as an emulsion in an acceptable oil, for example), ion exchange resins, or as sparingly soluble derivatives (such as a sparingly soluble salt, for example).

[0097] The compounds of this invention can also be administered in extended-release forms or extended-release drug delivery systems. A description of representative extended-release materials can also be found in the materials incorporated in Remington's Pharmaceutical Sciences. Administration mode

[0098] The compositions of this invention can be administered in any manner, including, but not limited to, oral administration, Petition 870250096065, dated 10 / 21 / 2025, pp. 52 / 90 / 51 parenteral, sublingual, buccal, nasal, intralesional, or combinations thereof. Parenteral administration includes, but is not limited to, subcutaneous and intramuscular routes. The compositions of this invention may also be administered in the form of an implant that allows slow release of the compositions as well as a slow controlled IV infusion. In a particular embodiment, one or more NOX4, NOX4 / 1, or NOX1 inhibitors are administered orally.

[0099] The dosage administered, whether single or multiple doses, to an individual will vary depending on a variety of factors, including pharmacokinetic properties, patient conditions and characteristics (age, body weight, health, body size), severity of symptoms, frequency of treatment, and the desired effect. Combination

[00100] According to one embodiment of the invention, a NOX4, NOX4 / 1 or NOX1 inhibitor according to the invention and pharmaceutical formulations thereof shall be administered in combination with an anticancer immunotherapeutic agent, in particular an anticancer vaccine or at least one immune checkpoint inhibitor such as at least one PD-1, PD-L1 or CTLA4 inhibitor.

[00101] The invention encompasses the administration of a NOX4, NOX4 / 1 or NOX1 inhibitor or a pharmaceutical formulation thereof, wherein the NOX4 / 1 inhibitor or a pharmaceutical formulation thereof is administered to an individual before or simultaneously with an anticancer immunotherapeutic agent, for example concomitantly via the same formulation or separately via different formulations, in particular via different formulation routes.

[00102] According to a particular aspect of the invention, a NOX4, NOX4 / 1 or NOX1 inhibitor according to the invention and pharmaceutical formulations thereof are to be administered chronically (by Petition 870250096065, dated 10 / 21 / 2025, page 53 / 90 / 51 example, daily or weekly) for the duration of treatment and before the administration of an anticancer immunotherapeutic agent or antiangiogenic treatment.

[00103] According to another particular aspect of the invention, a NOX4, NOX4 / 1 or NOX1 inhibitor according to the invention and pharmaceutical formulations thereof are to be administered concomitantly with an anticancer immunotherapeutic agent.

[00104] According to another particular aspect of the invention, the anticancer immunotherapeutic agent can be administered in combination with other therapeutic regimens or co-agents useful in the treatment of cancer (e.g., multiple drug regimens), in a therapeutically effective amount, such as in combination with substances useful for treating, stabilizing, preventing and / or delaying cancer, such as substances used in conventional chemotherapy directed against solid tumors and for controlling the establishment of metastases, or any other molecule that acts by triggering programmed cell death, for example, a co-agent selected from angiogenesis inhibitors (e.g., anti-VEGF agents as described in Gardner et al., 2017, supra), immunotherapy agents (e.g., recombinant cytokines, interferons, interleukins, recombinant antibodies such as Herceptin®) and chemotherapeutic agents (e.g., cisplatin, paclitaxel, methotrexate, 5-fluorouracil, gemcitabine, vincristine, vinblastine, doxorubicin, temozolomide). In particular, according to another particular aspect of the invention, the anticancer immunotherapeutic agent can be administered in combination with other therapeutic regimens or co-agents useful in the treatment of cancer (e.g., multiple drug regimens), in a therapeutically effective amount, such as in combination with at least one vascular endothelial growth factor (VEGF) inhibitor (e.g., bevacizumab, sunitinib inhibitors), at least one inhibitor of the factor of. Petition 870250096065, dated 10 / 21 / 2025, page 54 / 90 / 51 basic fibroblast growth factor (bFGF) inhibitor or at least one hypoxia-inducible factor 1 (HIF-1) inhibitor.

[00105] The NOX4 / 1 inhibitor or pharmaceutical formulations thereof that are administered concomitantly with said anticancer immunotherapeutic agent may be administered in or within the same composition(s) or distinct composition(s) and by the same route(s) or distinct route(s) of administration. Patients

[00106] In one embodiment, individuals according to the invention are individuals suffering from a solid tumor cancer, in particular an insufficiently responsive solid tumor cancer that exhibits or is likely to exhibit resistance to immunotherapy or an anti-angiogenic agent, in particular an anti-VEGF treatment.

[00107] In a particular embodiment, individuals according to the invention are individuals suffering from a selected solid tumor cancer of lung cancer (small cell and non-small cell), breast cancer, ovarian cancer, cervical cancer, uterine cancer, head and neck cancer, melanoma, hepatocellular carcinoma, colon cancer, rectal cancer, colorectal carcinoma, renal cancer, prostate cancer, gastric cancer, bronchial cancer, pancreatic cancer, urinary bladder cancer, liver cancer and brain cancer, in particular glioblastoma.

[00108] In a particular embodiment, individuals according to the invention are individuals suffering from a solid tumor cancer and have high expression of α-smooth muscle actin (α-SMA).

[00109] In another particular embodiment, individuals according to the invention are individuals suffering from hepatocellular carcinoma (HCC).

[00110] In another particular embodiment, individuals according to the invention are individuals suffering from head and neck tumors.

[00111] In another particular modality, individuals according Petition 870250096065, dated 10 / 21 / 2025, pp. 55 / 90 / 51, concerning the invention, refers to individuals suffering from melanoma.

[00112] In another particular embodiment, individuals according to the invention are individuals suffering from colon cancer.

[00113] In another particular embodiment, individuals according to the invention are individuals suffering from lung carcinoma.

[00114] In another particular embodiment, individuals according to the invention are individuals suffering from breast cancer.

[00115] In another particular embodiment, individuals according to the invention are individuals suffering from hepatocellular carcinoma or liver cancer.

[00116] In another particular embodiment, individuals according to the invention are individuals suffering from rectal cancer or colorectal carcinoma.

[00117] In another particular embodiment, individuals according to the invention are individuals suffering from kidney cancer.

[00118] In another particular embodiment, individuals according to the invention are individuals suffering from pancreatic cancer.

[00119] In another particular embodiment, individuals according to the invention are individuals suffering from brain cancer, in particular glioblastoma.

[00120] In another particular embodiment, individuals according to the invention are individuals with solid tumor cancer who are at risk of developing resistance or partial resistance to anticancer immunotherapy due to another concomitant treatment or a genetic predisposition.

[00121] In another particular embodiment, individuals according to the invention are individuals with hematological neoplasms such as lymphomas or leukemias. Use in accordance with the invention.

[00122] In a particular embodiment, the invention provides compounds, Petition 870250096065, dated 10 / 21 / 2025, page 56 / 90 / 51 methods, uses and compositions useful for the treatment of a solid tumor cancer in the form of a combination in which at least one NOX4 / 1 inhibitor must be administered in combination with at least one anticancer immunotherapeutic agent.

[00123] The references cited herein are hereby incorporated by reference in their entirety. The present invention shall not be limited in scope by the specific embodiments described herein, which provide unique illustrations of individual aspects of the invention, and functionally equivalent methods and components are within the scope of the invention. In fact, several modifications of the invention, in addition to those shown and described herein, will become apparent to those skilled in the art from the preceding description and accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

[00124] The invention having been described, the following examples are presented by way of illustration, and not limitation. EXAMPLES

[00125] The efficacy of NOX4 / 1 inhibitors in restoring or increasing responsiveness to an anticancer immunotherapeutic agent can be tested as follows: Example 1: Combination of NOX4 / 1 inhibitors and an anti-PD1 inhibitor in cancer treatment.

[00126] In order to test the effectiveness of a combination according to the invention, the following experiments are conducted in a mouse xenograft tumor model as described below.

[00127] Subcutaneous xenograft tumors composed of C38 cells (colon cancer), CT26 cells (colon cancer), LLC1 cells (lung carcinoma), B16F10 cells (melanoma), Hepa1-6 cells (liver cancer), or Renca cells (kidney cancer) are injected subcutaneously into the flank of C57Bl / 6 or Balb / c mice (2 to 3 months old). Alternatively, a Petition 870250096065, dated 10 / 21 / 2025, pp. 57 / 90 / 51 MC-38 cell lines derived from murine colonic adenocarcinoma cells C57BL6 or 4T1 mouse mammary tumor models are used.

[00128] Combination treatment begins when tumors reach an average volume of 80 to 200 mm3. Mice are randomized according to their individual tumor volume into different groups of 8 to 17 mice. Each group receives placebo or a NOX4 / 1 inhibitor alone or a PD-1 antibody alone or NOX4 / 1 in combination with PD-1 antibody.

[00129] The NOX4 / 1 inhibitors 2-(2-chlorophenyl)-4-[3(dimethylamino)phenyl]-5-methyl-1H-pyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione or (R)-3-methoxy-4-(2-morpholino-1-phenylethoxy)-N-(5-(pyridin-4-yl)-1,3,4thiadiazol-2-yl)benzamide are prepared daily (7 days / week) in 1.2% Methylcellulose plus 0.8% Polysorbate 80 (Sigma) and are administered to the animals of the respective groups by oral gavage via gavage tube at a dose of 60 and 10 mg / kg respectively.

[00130] As a PD-1 inhibitor, an anti-PD-1 antibody (ref.: BE0146, BioXcell; clone: ​​RMP1-14, reactivity: mouse; isotype: mouse IgG2a; storage conditions: +4°C) is injected into the peritoneal cavity of mice (Intraperitoneally, IP). The administration volume is 10 mL / Kg adjusted to the most recent individual body weight of the mice. Tumor collection and immunochemistry to assess T cell infiltration

[00131] Fourteen (14) days after randomization and if the antitumor activity of NOX4 / 1 compounds alone or in combination is considered sufficient, tumors from 5 satellite mice per group are collected, weighed and the tumor is cut into 2 fragments. One fragment is cut into 4 mm thick slices and joined in 4% neutral buffered formalin for 24 to 48h and then embedded in paraffin. Petition 870250096065, dated 10 / 21 / 2025, pp. 58 / 90 / 51 (Histosec®, Merck, Darmstadt, Germany). A fragment is embedded in Tissue Freezing Medium (Microm Microtech, France), instantly frozen in isopentane cooled in liquid nitrogen, and stored at 80°C until processing. Immunohistochemical staining for CD3, CD4, and CD8 is performed on paraffin-embedded tissue sections using standard techniques (Biodoxis, France). The number of CD3, CD4, and CD8 immunopositive cells per field is counted. Tumor collection and flow cytometry to assess T cell infiltration

[00132] Fourteen days after randomization, the tumor of 4 mice per group is collected.

[00133] All tumors are collected in RPMI culture medium (ref: BE12-702F, Lonza, Verviers, Belgium). Tumor-infiltrated immune cells are quantified by flow cytometry analysis of each collected sample. Then, antibodies directed against the chosen markers are added, according to the procedure described by the supplier for each antibody. All antibodies except FoxP3 will be for surface labeling and FoxP3 for intracellular labeling. The antibodies used for flow cytometry analysis for effector T-cell lymphocytes (Teff: CD45, CD3, CD8) and regulatory T-cell lymphocytes (Treg: CD45, CD3, CD4, FoxP3) in mouse samples are listed in Table 1 below: Table 1 Specificity and fluorochrome Reference Supplier Isotype and fluorochrome Isotype reference Supplier CD45 APCCy7 557659 BD Biosciences Rat IgG2bk APC-Cy7 552773 BD Biosciences CD3 V450 561389 BD Biosciences Rat IgG2bk V450 560457 BD Biosciences CD8 PerCP 553036 BD Biosciences Rat IgG2ak PerCP 553933 BD Biosciences FoxP3 PE 130-093014 Miltenyi Biotec PE Petition 870250096065, dated 10 / 21 / 2025, pp. 59 / 90 / 51 130-102CD4 Viogreen 444 Miltenyi Biotech IgG2b Viogreen 130-102- Miltenyi 659 Biotech

[00134] Stained cells are analyzed using a BD® LSR II flow cytometer (BD Biosciences) equipped with 3 excitation lasers at wavelengths of 405, 488, and 633 nm. Flow cytometry data are acquired until 10,000 mCD45+ events are recorded for each sample or for a maximum duration of 2 minutes. Animal monitoring

[00135] All study data, including measurements of animal body weight, tumor volume, clinical and mortality records, and treatment data, are scheduled and recorded. Viability and behavior are recorded daily. Body weights are measured twice weekly. Tumor length and width are measured twice weekly with calipers, and tumor volume is estimated using the formula: width 2 x length Tumor Volume =-----------2

[00136] Human endpoints. The experiment is terminated after 5 weeks or if: • The tumor exceeds 10% of normal body weight or exceeds 1,500 mm3 in mice, • Tumors interfere with movement or nutrition, > 8 mm of ulcerated tumor, hemorrhagic infection, • Tissue erosion, • 20% loss of remaining body weight by 2 days of monitoring (30% for one day of monitoring) compared to the day of treatment initiation / maximum weight, • Signs of pain, suffering or distress: pain posture, pain facial mask, behavior, • Poor body condition, emaciation, cachexia, dehydration, • Prolonged absence of voluntary responses to stimuli Petition 870250096065, dated 10 / 21 / 2025, page 60 / 90 / 51 external, • Rapid difficulty breathing, anemia, significant hemorrhage, • Neurological signs: spinning, seizure, paralysis, • Prolonged decrease in body temperature, • Abdominal distension. Effectiveness parameters

[00137] Treatment efficacy is evaluated in terms of the effects of the test substances on the tumor volumes of treated animals relative to control animals. The following criteria for evaluating antitumor efficacy are determined.

[00138] • Individual and / or average (or median) tumor volumes will be provided, • Tumor doubling time (TD) will be calculated, • Tumor growth inhibition (% T / C) defined as the ratio of average tumor volumes of the treated group versus the control group will be calculated: Average tumor volume of the treated group in DX T / C % =-------------------—-------------x 100 Average tumor volume of the vehicle-treated group in DX

[00139] The optimal value is the minimum % T / C ratio reflecting the maximum tumor growth inhibition achieved. The effective criteria for the % T / C ratio according to NCI standards is * 42%. The volume V and the time to reach V are calculated. The volume V is defined as a target volume deduced from experimental data and chosen in the exponential phase of tumor growth. For each tumor, the tumor volume closest to the target volume V is selected from the tumor volume measurements. The value of this volume V and the time for the tumor to reach this volume are recorded. For each group, the average tumor volumes V and the average times to reach this volume are calculated. Mouse survival will also be monitored and used as an efficacy parameter. Survival curves are plotted. Petition 870250096065, dated 10 / 21 / 2025, pp. 61 / 90 / 51

[00140] When MC38 cancer cells (0.5 x 105) are used, they are injected subcutaneously (sc) in phosphate-buffered saline (PBS) into the flank of female mice aged 8 to 10 weeks. MC38 cells are injected alone or mixed with C57BL / 6 colonic fibroblasts (2.5 x 105), pre-treated ex vivo prior to injection with 2 ng / ml of TGFe1 for 6 days to induce a CAF phenotype.

[00141] When 4T1 cancer cells (0.5 x 105) are used, they are injected sc in PBS into the upper mammary fat pad of female mice aged 8 to 10 weeks. The cells are injected either alone or mixed with 2.5 x 10⁵ CAFs of mammary BALB / C isolated from spontaneous transgenic BALBneuT stroma-rich mammary tumors.

[00142] The NOX4 inhibitor 2-(2-chlorophenyl)-4-[3-(dimethylamino)phenyl]-5-methyl-1H-pyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione (GKT137831) was administered to mice when tumors were palpable. GKT137831 was reconstituted in 1.2% Methyl Cellulose (Sigma) with 0.1% Polysorbate (Sigma) and administered by oral gavage 5 times / week at 40 mg / kg. Control mice received vehicle by oral gavage. For long-term dosing, 15 initial doses were given as established, but reduced to 3 times / week for 3 weeks at 50 mg / kg, then 2 times / week for 3 weeks at 60 mg / kg. The anti-PD-1 antibody (Bioxcell; RMP1-14) was administered via intraparietal (ip) injection. 300 μg of the antibody or the IgG2a isotype control (Bioxcell) were given when tumors were palpable on alternate days, totaling 3 doses.

[00143] For the data presented in Figure 1, tumors were measured every 2-3 days using an electronic skin caliper based on their longest width and length. Tumor volume was calculated using the formula 4 / 3nXr3, where the radius (r) was calculated from the measurement of the tumor's width and length to provide an average diameter value. The Petition 870250096065, dated 10 / 21 / 2025, page 62 / 90 / 51 mice were randomized into groups based on tumor volume so that no statistical difference occurred between the mean tumor volumes between the groups before treatments began. Figure 1A shows that on day 15, that is, after 8 days of treatment, the tumors were significantly smaller when mice were treated with the NOX4 inhibitor than compared to vehicle alone. Furthermore, immunochemistry (performed as described above) revealed, as represented in Figures 1B and 1C, respectively, that treatment with the NOX4 inhibitor significantly reduces CAF-positive SMA in tumors and results in the relocation of CD8+ T cells from the tumor edge to the tumor center.Using the 4T1 breast cancer model, these results clearly show that GKT treatment inhibits CAF formation, as demonstrated by the decreased myofibroblast population (SMA-positive cells), allowing CD8+ T cells to access the tumor and eliminate cancerous cells, reducing tumor size. This supports the beneficial effects of combining a NOX4 inhibitor and an anticancer immunotherapeutic agent that would further activate CD8+ T cells.

[00144] The beneficial effects of such a combination are further supported by the results presented in Figure 2 for the combination of a PD-1 inhibitor (aPD1) with the NOX4 inhibitor GKT137831, which significantly improves the therapeutic response in CAF-rich tumors: tumors were significantly smaller when mice were treated with the aPD1 / GKT831 combination compared to aPD1 alone (Figure 2A), and following administration of the aPD1 / GKT831 combination, there is a significant relocation of CD8+ T cells from the tumor edge to the tumor center (Figure 2B), and the survival outcome is also significantly increased (Figure 2C) compared to aPD1 alone. Using the MC38 colon cancer model, the beneficial effect of the Petition 870250096065, dated 10 / 21 / 2025, pp. 63 / 90 / 51: GKT / aPD1 combination therapy was confirmed, showing a very significant decrease in tumor volume, which is accompanied by an increase in mouse survival. Furthermore, this effect was shown to result from CD8+ T cell infiltration within the tumor by NOX inhibitors. These results strongly suggest that the NOX inhibitors of the invention, in particular GKT137831, are strong candidates for co-therapy with PD1 for all CAF-rich cancers. Example 2: Combination of NOX4 / 1 inhibitors and a cancer vaccine in cancer treatment.

[00145] In order to test the effectiveness of a combination according to the invention, NOX4 / 1 inhibitors are combined with treatment with a vaccine such as an anti-HPV vaccine.

[00146] TC1 (0.5 x 105) cancer cells (prostate cancer) were injected subcutaneously (sc) in phosphate-buffered saline (PBS) into the flank of female mice aged 8–10 weeks. TC1 cells were injected alone or mixed with C57BL / 6 (2.5 x 105) lung fibroblasts, pretreated ex vivo prior to injection with 2 ng / ml of TGFe1 for 6 days to induce a CAF phenotype.

[00147] Tumors were measured every 2–3 days using an electronic skin caliper based on their longest width and length. Tumor volume measurements, randomized mice, and dosage by oral gavage were performed as described above.

[00148] Vaccination with a DNA vaccine encoding tetanus Fragment C domain 1 (Dom) fused to the immunodominant CD8 epitope of HPV E7 RAHYNIVTF (RAH, E749-57) (Rice et al. 2002, J. Immunol., 169: 3908-13; Rice et al., 2008, Nat Rev Cancer, 8: 108-20) was administered via intramuscular (im) injection when tumors were palpable. One injection containing 50 μg of DNA in PBS was given, and any repeat doses were given 3 weeks after the initial immunization. Treatment with an inhibitor of Petition 870250096065, dated 10 / 21 / 2025, pp. 64 / 90 / 51 NOX4 (GKT137831), reconstituted as described in Example 1, was administered to mice when tumors were palpable.

[00149] Figure 3 supports the idea that combining an antitumor vaccination with a NOX4 inhibitor significantly improves the therapeutic response in CAF-rich tumors, as on day 24 the tumors were significantly smaller when mice were treated with the vaccine / NOX4 inhibitor combination compared to the vaccine alone. Following administration of the vaccine / NOX4 inhibitor combination, there was a significant relocation of CD8+ T cells from the tumor edge to the tumor center (Figure 3B), and the survival outcome was also significantly increased (Figure 3C) compared to the vaccine alone. Effective immunotherapy, whether based on checkpoint inhibitors, T-cell agonists, vaccination, or adoptive T-cell transfer, requires the presence of effector CD8+ T cells in the tumor.Cancer-associated fibroblasts are found in most solid cancers and play a major role in tumor immune evasion by excluding CD8+ T cells from cancers, thus rendering immunotherapies ineffective. Therefore, since the NOX inhibitors of the invention, in particular GKT831, effectively target CAF as shown by the decrease in SMA-positive cells in the 4T1 model, they promote CD8+ T cell infiltration within tumors and restore the response to vaccine-based and PD1-based immunotherapies. These data suggest that combination immunotherapy with NOX4 inhibitors of the invention, in particular GKT137831, can significantly improve response rates for this type of treatment. Example 3: Combination of NOX4 / 1 inhibitors and antiVEGF agent in cancer treatment.

[00150] In order to test the effectiveness of a combination according to the invention, NOX4 / 1 inhibitors are combined with treatment with a Petition 870250096065, dated 10 / 21 / 2025, p. 65 / 90 / 51 antiVEGF agent.

[00151] Mouse models of MC38 tumor xenografts were produced by injecting PBS-diluted MC38 tumor cells (5x10⁵ for MC38) subcutaneously into wild-type C57 / BL6 mice or NOX1-deficient mice (NOX1-KO). When tumors reached 50 mm³, intraperitoneal administration of purified antibodies: an anti-VEGF:DC101 or an irrelevant mouse IgG (as a control) was performed twice weekly. DC101 was given at a dose of 600 µg per injection per mouse. Vehicle (VL) (i.e., methylcellulose and Tween 80) or a selective NOX1 inhibitor, (R) 3-methoxy-4(2-morpholino-1-phenylethoxy)-N-(5-(pyridin-4-yl)-1,3,4-thiadiazol-2-yl)benzamide (GKT2) (twice daily at 10 mg / kg) were given by oral gavage until sacrifice of the mice. Tumor size was measured with a caliper and tumor volume was determined according to the equation: (Length*width*thickness).Tumor size was measured in vivo using a caliper (D-0 to D-15) every 5 days. After sacrifice, tumors were removed without fixation with PFA (paraformaldehyde), isolated, and blood vascular endothelial cells (CD45- / CD31+ / GP38-) were analyzed by flow cytometry.

[00152] Figure 4 shows that the combination of a highly selective NOX1 inhibitor (GKT2) and an anti-VEGF-R2 blocking antibody (DC101) allows for the inhibition of angiogenesis. Furthermore, GKT2 and DC101 act synergistically in enhancing the inhibition of neovascularization.

[00153] Figure 5 shows that tumors in NOX1KO mice exhibited decreased growth kinetics compared to tumors in WT mice, indicating a clear involvement of NOX1. Furthermore, treatment with the anti-VEGFR2 antibody (DC101) decreased tumor growth in NOX1-deficient mice, and this effect was even more pronounced compared to wild-type mice. This Petition 870250096065, dated 10 / 21 / 2025, pp. 66 / 90 / 51 clearly suggests different mechanisms of action between VEGFR2 and NOX1 signaling.

[00154] Therefore, overall, these data support the idea that combining NOX1 inhibition with antiangiogenic agents such as antiVEGF inhibitors would allow for a synergistic effect in tumor treatment. Petition 870250096065, dated 10 / 21 / 2025, pp. 67 / 90

Claims

1 / 3 CLAIMS 1. Use of 2-(2-chlorophenyl)-4-[3-(dimethylamino)phenyl]-5-methyl-1H-pyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione, characterized in that it is for the manufacture of a medicament for treating solid tumor cancers exhibiting or susceptible to exhibiting resistance to immunotherapy, wherein said 2-(2-chlorophenyl)-4-[3-(dimethylamino)phenyl]-5-methyl-1H-pyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione is to be administered in combination with an anticancer immunotherapeutic agent selected from at least one cancer vaccine or at least one immune checkpoint inhibitor.

2. Use according to claim 1, characterized in that said 2-(2-chlorophenyl)-4-[3-(dimethylamino)phenyl]-5-methyl-1H-pyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione is to be administered in combination with at least one immune checkpoint inhibitor.

3. Use according to claim 2, characterized in that said 2-(2-chlorophenyl)-4-[3-(dimethylamino)phenyl]-5-methyl-1Hpyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione is to be administered in combination with a PD-1 inhibitor.

4. Use according to claim 2, characterized in that said 2-(2-chlorophenyl)-4-[3-(dimethylamino)phenyl]-5-methyl-1Hpyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione is to be administered in combination with a CTLA-4 inhibitor.

5. Use according to claim 1, characterized in that said 2-(2-chlorophenyl)-4-[3-(dimethylamino)phenyl]-5-methyl-1Hpyrazolo[4,3-c]pyridine-3,6(2H,5H)-dione is to be administered in combination with a cancer vaccine selected from oncolytic and Herpes simplex antiviral vaccines.

6. Use in accordance with any of claims 1 to 5, Petition 870250096065, dated 10 / 21 / 2025, p. 68 / 90 2 / 3 characterized in that said cancer vaccine is a Herpes simplex antiviral vaccine that is T-Vec (Imlygic, talimogene laherparepvec), or said immune checkpoint inhibitor is a PD-1 inhibitor selected from Pembrolizumab (Keytruda) and Nivolumab (Opdivo), or said immune checkpoint inhibitor is a CTLA-4 inhibitor that is Ipilimumab (Yervoy).

7. Use in accordance with any of the preceding claims, characterized in that the solid tumor cancer is selected from lung cancer (small cell and non-small cell), breast cancer, ovarian cancer, cervical cancer, uterine cancer, head and neck cancer, melanoma, hepatocellular carcinoma, colon cancer, rectal cancer, colorectal carcinoma, renal cancer, prostate cancer, gastric cancer, bronchial cancer, pancreatic cancer, urinary bladder cancer, liver cancer and brain cancer, in particular glioblastoma.

8. Use in accordance with any of the preceding claims, characterized in that the solid tumor cancer is selected from lung cancer, breast cancer, head and neck cancer, colorectal carcinoma, prostate cancer, and pancreatic cancer.

9. Pharmaceutical composition, characterized in that it contains 2-(2-chlorophenyl)-4-[3-(dimethylamino)phenyl]-5-methyl-1H-pyrazolo[4,3c]pyridine-3,6(2H,5H)-dione combined with at least one anticancer immunotherapeutic agent and at least one pharmaceutically acceptable carrier, wherein said at least one anticancer immunotherapeutic agent is selected from at least one cancer vaccine or at least one immune checkpoint inhibitor.

10. Pharmaceutical formulation according to claim 9, characterized in that the anticancer immunotherapeutic agent is a cancer vaccine such as Herpes simplex antiviral vaccines.

11. Use in accordance with any of claims 1 to 3 and 6 to 8, characterized in that the anticancer immunotherapeutic agent is a PD-1 inhibitor selected from Pembrolizumab (Keytruda) and Nivolumab (Opdivo).

12. Pharmaceutical formulation according to claim 9, characterized in that the anticancer immunotherapeutic agent is a PD-1 inhibitor selected from Pembrolizumab (Keytruda) and Nivolumab (Opdivo).

13. Use in accordance with any of claims 1, 2, 4 and 6 to 8, characterized in that the anticancer immunotherapeutic agent is a CTLA-4 inhibitor, which is Ipilimumab (Yervoy).

14. Pharmaceutical formulation according to claim 9, characterized in that the anticancer immunotherapeutic agent is a CTLA-4 inhibitor, which is Ipilimumab (Yervoy). Petition 870250096065, dated 10 / 21 / 2025, pp. 70 / 90

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