Cancer homeostasis disruption technology

Abstract

A method of treating a disease in a subject, selected from cancer, cancer-associated inflammation, chronic inflammatory disorders, autoimmune diseases and age-associated systemic inflammation and / or combinations, comprising administering to the subject a therapeutically effective amount of a compound of formula (I), wherein G is an L- or D-glucose C-glycosyl pyranose residue bearing hydroxyl or OR7, SR7, NR8R9, O-C(=O)R10 or O-C(=O)OR10 substituents at C-2, C-3, C-4 and C-6, the anomeric C-1 bonded via a divalent linker L to an –NH–CO–AA moiety; L is a C1–6 alkylene optionally containing O, S or NRᵃ; AA is a residue of a basic amino acid selected from lysine, arginine, ornithine, homoarginine, histidine, diaminobutyric acid and diaminopropionic acid or non-naturally occurring analogs thereof having a side-chain pKₐ ≥7; and including any enantiomer, diastereomer, racemate, pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, thereby modulating an abnormal intracellular-to-extracellular pH gradient in diseased cells.

Description

Docket No.: 411037-501001 WOCANCER HOMEOSTASIS DISRUPTION TECHNOLOGYCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to United States (US) provisional application Serial No. 63 / 730,058, filed December 10, 2024, the entire disclosure of which is incorporated by reference as if fully set forth herein in its entirety.FIELD OF THE INVENTION

[0002] The present disclosure relates to a method of treating diseases, and more particularly, relates to a method of treating diseases such as cancer, chronic inflammatory disorders, and autoimmune diseases by strategically modulating pH gradients in diseased cells.BACKGROUND OF THE INVENTION

[0003] Cancer has been recognized as a distinct set of diseases for thousands of years. Descriptions of ulcerating masses and progressive “crab-like” lesions appear in writings from ancient Egypt and classical Greece, and the term “cancer” itself derives from the Greek word “karkinos” used by Hippocrates. Over subsequent centuries, advances in anatomy and pathology established that malignant tumors arise from the uncontrolled proliferation of the body’s own cells, which can invade surrounding tissues and spread to distant organs.

[0004] During the twentieth century, the modern understanding of cancer as a genetic and cellular disease emerged. Malignant cells were shown to acquire multiple abnormalities that promote sustained proliferation, resistance to cell death, angiogenesis, invasion and metastasis. Large bodies of work defined cancer as a collection of related diseases characterized by progressive accumulation of mutations, epigenetic alterations and dysregulated signaling pathways rather than as a single entity.

[0005] Historically, therapeutic efforts have focused on eliminating or controlling malignant cells once they arise. The earliest widely adopted modality was surgical excision, which remains a cornerstone of therapy for many localized solid tumors. With the development of ionizing radiation, radiotherapy became a second major pillar of treatment, allowing local control of tumors that could not be completely resected.

[0006] Systemic drug treatment was introduced in the mid-twentieth century with the advent of cytotoxic chemotherapeutic agents. These agents typically interfere with DNA replication, mitotic spindle formation or other processes essential for rapidly dividing cells. Classic examples include antimetabolites such as 5-fluorouracil and microtubule-targeting agents such as taxanes. Although such drugs can produce substantial tumor regression andDocket No.: 411037-501001 WO long-term control in some settings, their lack of selectivity for malignant over normal proliferating cells is associated with significant toxicity, including bone marrow suppression, gastrointestinal side effects and hair loss.

[0007] More recently, the identification of recurrent molecular alterations in specific tumor types has led to the development of targeted therapies. These agents are designed to inhibit oncogenic kinases, hormone receptors or other defined molecular drivers in subsets of patients. In parallel, immunotherapy has become an important treatment modality, particularly with the advent of monoclonal antibodies and immune checkpoint inhibitors that enhance endogenous anti-tumor immune responses. Examples include antibodies directed against growth factor receptors and antibodies that block inhibitory receptors on T cells.

[0008] Despite these advances, cancer remains a leading cause of morbidity and mortality worldwide. Many patients present with advanced or metastatic disease that is not amenable to curative surgery or radiotherapy. Tumors often exhibit substantial genetic and phenotypic heterogeneity, and cancer cells can adapt to and escape from selective pressures imposed by therapy. Clinical problems such as primary resistance, acquired resistance, relapse after an initial response and treatment-limiting toxicities continue to restrict the effectiveness of existing therapeutic options.

[0009] There is therefore an ongoing need for further improvements in the prevention and treatment of cancer and related proliferative disorders, including approaches that may be used alone or in combination with existing modalities to improve patient outcomes.BRIEF SUMMARY OF THE INVENTION

[0010] For purposes of summary, certain aspects, advantages, and novel features are described herein. It is to be understood that not necessarily all such advantages may be achieved in accordance with any one particular embodiment. Thus, the apparatuses or methods claimed may be embodied or carried out in a manner that achieves or optimizes one advantage or group of advantages as taught herein without necessarily achieving other advantages as may be taught or suggested herein.

[0011] According to some aspects, the embodiments of the present invention provide methods of treating cancer and inflammatory disorders using glucose c-glycosyl basic amino acid compounds.

[0012] In some embodiments, the present disclosure relates to small-molecule modulators of cellular and microenvironmental pH that differ fundamentally from classical inhibitors of ion exchangers, proton pumps, or monocarboxylate transporters. Traditional Na+ / H+exchanger (NHE) inhibitors, vacuolar-type H+-ATPase (V-ATPase) inhibitors, proton pump inhibitors, and monocarboxylate transporter (MCT) inhibitors seek to impede proton or lactate efflux and thereby indirectly influence intracellular and extracellular pH. However,Docket No.: 411037-501001 WO such agents often exhibit limited isoform selectivity, heterogeneous uptake across tissues, and pharmacokinetic or safety profiles that constrain sustained dosing in oncology and inflammatory indications. According to some aspects of the present disclosure, the disclosed compounds are designed to engage the cancer cell pH gradient by an alternative strategy that does not rely exclusively on blockade of a single transporter or pump.

[0013] In some embodiments, the compounds comprise a glucose C-glycosyl residue covalently linked through a glycosidase-resistant carbon-carbon bond to a linker segment L and an amino acid or peptide segment AA, thereby providing constructs of formula (I) G-L-NH-CO-AA. The glucose C-glycosyl residue is configured to be recognized by facilitative glucose transporters and related uptake systems that are frequently upregulated in malignant and metabolically stressed cells. In contrast to O-glycosidic conjugates that undergo rapid enzymatic or chemical cleavage, the C-glycosidic linkage of the disclosed constructs provides enhanced stability in extracellular fluids and within intracellular compartments and thus supports sustained exposure of tumor cells to the pH- modulating scaffold. According to some aspects, the linker segment L is selected to control spatial orientation, protonation state, and membrane interaction of the conjugate, while the amino acid or peptide segment AA provides one or more ionizable groups that participate in proton capture, buffering, or shuttling within the cancer cell microenvironment.

[0014] In a non-limiting example, the intracellular compartment of a non-transformed cell maintains a slightly acidic to neutral pH, whereas the extracellular milieu is slightly alkaline. Many cancers exhibit a reversed gradient, characterized by relatively alkaline intracellular pH and markedly acidic extracellular pH created by excess proton and lactate export. According to some aspects, compounds of formula (I) are configured to interact with this aberrant gradient by binding, sequestering, or redistributing protons in a manner that drives the system toward physiological homeostasis. For example, protonatable side chains or peptide segments within the AA moiety may accumulate within acidic extracellular regions, accept protons, and subsequently alter proton flux across cellular and organellar membranes. Such activity has the potential to lower intracellular pH toward normal values while reducing extracellular acidity, thereby undermining metabolic adaptations that support proliferation, invasion, and resistance to therapy.

[0015] In some embodiments, the disclosed compounds exploit the combination of sugar-mediated uptake and pH-responsive functionality to achieve preferential action in cancer cells and inflamed tissues without reliance on extensive structural modification of existing chemotherapeutic agents. The glucose C-glycosyl residue can enhance delivery to cells with increased glucose consumption, while the amino acid or peptide segment may be tuned to modulate hydrophilicity, charge distribution, and subcellular localization. In certain implementations, basic amino acids, polycationic peptides, or amphipathic motifs within AADocket No.: 411037-501001 WO contribute to cell penetration and accumulation in acidic microenvironments, yet the overall construct remains resistant to rapid metabolic degradation due to the C-glycosidic linkage. According to some aspects, such modular design enables independent optimization of targeting, pH-modulating capacity, and safety profile.

[0016] In this example, experimental evaluation in breast cancer cell models, including monolayer and three-dimensional spheroid cultures, demonstrates that representative compounds of formula (I) can reduce tumor cell viability, modify intracellular and pericellular pH, and activate apoptotic pathways. Quantitative assays of cell survival, pH-sensitive fluorogenic probes, and caspase-3 / 7 activation indicate that treatment with such compounds induces collapse of the cancer-associated pH gradient and promotes programmed cell death while preserving key features of surrounding architecture. The figures of the present application illustrate non-limiting examples of these effects, including dose-response curves, changes in fluorescence intensity corresponding to pH shifts, and imaging of spheroid integrity and apoptosis markers.

[0017] According to some aspects, modulation of the tumor microenvironment by the disclosed compounds restores conditions that favor normal cellular homeostasis, increases vulnerability of malignant cells to intrinsic and extrinsic regulatory mechanisms, and provides a platform for combination regimens with chemotherapeutic, immunotherapeutic, or antiinflammatory agents.

[0018] As such, keeping in mind possible combination embodiments and the above brief discussion, as an additional brief summary or to provide discussion points for a brief summary, some example features contemplated for the technology disclosed herein can be briefly summarized by the following list of features, any of which can be inter-combined or discussed optionally with any other example, feature, Figure, Drawing, detail, embodiment, and / or aspect disclosed herein:

[0019] Feature 1: A method of treating a disease in a subject in need thereof, the disease being selected from cancer, cancer-associated inflammation, chronic inflammatory disorders, autoimmune diseases and age-associated systemic inflammation, and combinations thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I):Docket No.: 411037-501001 WO(L-glucose),

[0020] wherein: G is an L-glucose C-glycosyl residue or a D-glucose C-glycosyl residue in the pyranose form having the carbon numbering shown for formula (I), said residue having a six-membered ring with a -CH2OH substituent at C-5 and hydroxyl groups at C-2, C-3, C-4 and C-6, each of the hydroxyl groups at C-2, C-3, C-4 and C-6 being independently present as an unsubstituted hydroxyl group or as a group OR7, SR7, NR8R9, O-C(=O)R10or O-C(=O)OR10, and the anomeric carbon C-1 being bonded through a carboncarbon bond B1 to a carbon atom of the linker group L; R7is hydrogen, Ci-C4alkyl, C2-C6alkenyl, C2-C6alkynyl, benzyl, phenyl or phenyl substituted by one or more substituents selected from Ci-C4alkyl, Ci-C4alkoxy, halogen, hydroxy and nitro; R8and R9are independently hydrogen or Ci-C4alkyl; R10is Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, benzyl or phenyl; L is a divalent linker group connecting the anomeric carbon (C-1) of G to the nitrogen atom of the -NH-CO- group and is selected from Ci-C6straight or branched alkylene and C2-C6alkylene optionally interrupted by one or more heteroatoms selected from O, S and NRa, wherein L contains from 1 to 8 carbon atoms in total and Rais hydrogen or Ci-C4alkyl; AA is the residue of a basic amino acid of formula -NH-CHR11-(CH2)P-NR2R3- CO-, which residue is amide-bonded through its a-carboxyl group to the -NH-CO- moiety ofDocket No.: 411037-501001 WO formula (I), wherein p is an integer from 2 to 5; R11is hydrogen, Ci-C6alkyl optionally substituted by one or more substituents selected from hydroxy, amino, carboxy, Ci-C4alkoxycarbonyl and halogen, or is -(CH2)m-NR4R5where m is 1-4 and R4and R5are independently hydrogen or Ci-C4alkyl; R2and R3are independently hydrogen, Ci-C4alkyl, hydroxy-Ci-C4alkyl or a group -C(=NH)NH2, or R2and R3together with the nitrogen atom to which they are attached form a saturated 5-, 6- or 7-membered heterocycle optionally containing an additional nitrogen atom; with the proviso that AA is a residue of a basic amino acid selected from lysine, arginine, ornithine, homoarginine, histidine, diaminobutyric acid and diaminopropionic acid, or a non-naturally occurring analog thereof having a side-chain pKaof at least 7; and wherein the method includes administration of any enantiomer, diastereomer, racemate, pharmaceutically acceptable salt, solvate, hydrate or prodrug of the compound of formula (I), thereby modulating an abnormal intracellular-to-extracellular pH gradient in diseased cells in the subject.

[0021] Feature 2: The method of feature 1 , wherein the disease is cancer.

[0022] Feature 3: The method of feature 2, wherein the cancer is a solid tumor selected from breast cancer, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, ovarian cancer and glioblastoma.

[0023] Feature 4: The method of feature 1 , wherein the disease is a chronic inflammatory or autoimmune condition selected from rheumatoid arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, psoriasis, psoriatic arthritis, systemic lupus erythematosus and multiple sclerosis.

[0024] Feature 5: The method of any one of features 1-4, wherein administration of the compound of formula (I) increases extracellular pH and / or decreases intracellular pH in diseased cells so that the intracellular-to-extracellular pH gradient approaches that of corresponding non-malignant cells.

[0025] Feature 6: The method of any one of features 1-5, wherein the modulation of the pH gradient induces apoptosis in diseased cells, as evidenced by activation of caspase- 3 / 7 and loss of clonogenic survival.

[0026] Feature 7: The method of any one of features 1-6, wherein restoration of a physiological pH gradient enhances anti-tumor immune surveillance, including increased infiltration and cytotoxic activity of CD8+T cells and / or natural killer cells.

[0027] Feature 8: The method of any one of features 1-7, wherein the compound of formula (I) is administered to the subject orally, intravenously, intraperitoneally, intratumorally, peri-tumorally or by a combination of such routes.Docket No.: 411037-501001 WO

[0028] Feature 9: The method of any one of features 1-8, wherein the compound of formula (I) is administered at a dose in the range of about 0.01 to about 100 mg / kg body weight per day.

[0029] Feature 10: The method of any one of features 1-9, further comprising administering to the subject at least one additional therapeutic agent selected from a cytotoxic chemotherapeutic agent, a targeted small-molecule kinase inhibitor, an immune checkpoint inhibitor, a PARP inhibitor, a hormone or endocrine therapy agent, a monoclonal antibody or antibody-drug conjugate, an epigenetic agent or modulator, and a cell-based immunotherapy.

[0030] Feature 11 : The method of any one of features 1-10, further comprising coadministering to the subject at least one reverse transcriptase inhibitor (RTI), optionally a nucleoside or nucleotide RTI selected from lamivudine, censavudine and elvucitabine.

[0031] Feature 12: The method of any one of features 1-11, wherein L is -CH2-CH2-.

[0032] Feature 13: The method of any one of features 1-12, wherein AA is in the L- configuration at the a-carbon and is the residue of lysine or arginine.

[0033] Feature 14: The method of any one of features 1-13, wherein the compound of formula (I) is a compound selected from the group consisting of [structure A] and [structure B]:, [structure A], orDocket No.: 411037-501001 WO, [structure B]; wherein R is selected from H, Ci-6alkyl, Ci-6hydroxyalkyl, Ci-4fluoroalkyl, C3-6cycloalkyl, benzyl, phenethyl, phenyl, substituted phenyl, heteroaryl, and / or pyridyl, thienyl, or furyl.

[0034] Feature 15: The method of any one of features 1-14, wherein at least one of the hydroxyl groups at C-2, C-3, C-4 and C-6 is acylated as an O-C(=O)R10or O-C(=O)OR10group to provide a I ipophi lically masked prodrug.

[0035] Feature 16: The method of any one of features 1-15, for reducing the risk of tumor recurrence or metastatic spread in a subject who has previously received a primary cancer therapy comprising surgery, chemotherapy, radiotherapy, immunotherapy or a combination thereof.

[0036] Feature 17: A compound of formula (I): G-L-NH-CO-AA as defined in feature 1 , for use in therapy.

[0037] Feature 18: The compound for use according to feature 17, for use in treating cancer in a subject in need thereof.

[0038] Feature 19: The compound for use according to feature 18, for use in treating a cancer characterized by extracellular acidification and intracellular alkalinization relative to corresponding non-malignant cells.

[0039] Feature 20: The compound for use according to any one of features 17-19, for use in treating cancer-associated inflammation in a subject having a solid tumor.

[0040] Feature 21 : The compound for use according to any one of features 17-20, for use in treating a chronic inflammatory or autoimmune disease characterized by an acidic extracellular microenvironment at a site of inflammation.

[0041] Feature 22: The compound for use according to any one of features 17-21 , for use in enhancing anti-tumor immune surveillance by normalizing extracellular pH in a tumor microenvironment.

[0042] Feature 23: The compound for use according to any one of features 17-22, for use in preventing or reducing metastatic dissemination of a solid tumor.Docket No.: 411037-501001 WO

[0043] Feature 24: The compound for use according to any one of features 17-23, for use in preventing tumor recurrence following a primary cancer therapy.

[0044] Feature 25: The compound for use according to any one of features 17-24, for use in combination therapy with at least one immune checkpoint inhibitor selected from anti-PD-1 , anti-PD-L1 and anti-CTLA-4 antibodies.

[0045] Feature 26: The compound for use according to any one of features 17-25, for use in combination therapy with at least one reverse transcriptase inhibitor (RTI) to treat age-associated inflammation or cancer-associated inflammation.

[0046] Feature 27: The compound for use according to any one of features 17-26, for use in combination with radiotherapy, wherein the compound of formula (I) is administered so as to reverse an abnormal intracellular-to-extracellular pH gradient and improve the response of the cancer to ionizing radiation.

[0047] Feature 28: A pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I): G-L-NH-CO-AA as defined in feature 1, and at least one pharmaceutically acceptable carrier, diluent or excipient.

[0048] Feature 29: The pharmaceutical composition of feature 28, formulated for oral administration and in a solid dosage form selected from a tablet, capsule, granule and powder.

[0049] Feature 30: The pharmaceutical composition of feature 28, formulated for intravenous administration and in a dosage form selected from a sterile aqueous solution, a sterile aqueous suspension and a lyophilized powder for reconstitution.

[0050] Feature 31 : The pharmaceutical composition of feature 28, formulated for local administration to a tumor or tumor bed and in a dosage form selected from an injectable solution, an injectable suspension, a gel, an in s / tu-forming depot, an implant and a biodegradable polymer matrix.

[0051] Feature 32: The pharmaceutical composition of any one of features 28-31 , wherein the compound of formula (I) is present as a pharmaceutically acceptable salt, solvate, hydrate or prodrug.

[0052] Feature 33: The pharmaceutical composition of any one of features 28-32, further comprising at least one additional therapeutic agent selected from the additional therapeutic agents recited in feature 10.

[0053] Feature 34: The pharmaceutical composition of feature 33, wherein the additional therapeutic agent is a cytotoxic chemotherapeutic agent selected from doxorubicin, paclitaxel, carboplatin and combinations thereof.

[0054] Feature 35: The pharmaceutical composition of any one of features 28-34, further comprising at least one immune checkpoint inhibitor selected from nivolumab, pembrolizumab, atezolizumab and ipilimumab.Docket No.: 411037-501001 WO

[0055] Feature 36: The pharmaceutical composition of any one of features 28-35, further comprising at least one reverse transcriptase inhibitor selected from lamivudine, censavudine and elvucitabine.

[0056] Feature 37: The pharmaceutical composition of any one of features 28-36, wherein the composition is in unit dosage form containing from about 1 mg to about 500 mg of the compound of formula (I).

[0057] Feature 38: The pharmaceutical composition of any one of features 28-37, wherein the composition is formulated as a controlled-release or sustained-release dosage form adapted to provide plasma levels of the compound of formula (I) sufficient to modulate a cancer-associated pH gradient over a period of at least 8 hours.

[0058] Feature 39: A kit comprising (a) a pharmaceutical composition according to any one of features 28-38 and (b) printed instructions for use of the composition in the treatment of a disease as defined in feature 1.

[0059] Feature 40: A pharmaceutical combination comprising: (a) a compound of formula (I): G-L-NH-CO-AA as defined in feature 1 ; and (b) at least one reverse transcriptase inhibitor (RTI); wherein components (a) and (b) are formulated or packaged for concurrent, sequential or separate administration to a subject in need thereof.

[0060] Feature 41 : The pharmaceutical combination of feature 40, wherein the at least one RTI is a nucleoside or nucleotide RTI.

[0061] Feature 42: The pharmaceutical combination of feature 41 , wherein the at least one RTI is selected from lamivudine, emtricitabine, tenofovir, abacavir, zidovudine and combinations thereof.

[0062] Feature 43: The pharmaceutical combination of any one of features 40-42, wherein the at least one RTI is selected from censavudine, elvucitabine and combinations thereof.

[0063] Feature 44: The pharmaceutical combination of any one of features 40-43, further comprising at least one additional therapeutic agent selected from a cytotoxic chemotherapeutic agent, a targeted kinase inhibitor and an immune checkpoint inhibitor.

[0064] Feature 45: The pharmaceutical combination of any one of features 40-44, for use in treating cancer or a cancer-associated inflammatory disorder in a subject in need thereof.

[0065] Feature 46: The pharmaceutical combination of any one of features 40-44, for use in treating an age-associated or chronic inflammatory condition characterized by an acidic microenvironment.

[0066] Feature 47: The pharmaceutical combination of any one of features 40-46, wherein the compound of formula (I) is administered prior to or concurrently with the at leastDocket No.: 411037-501001 WO one RTI so as to normalize extracellular pH and reduce RTI-sensitive inflammatory signaling within a tumor microenvironment.

[0067] Feature 48: The pharmaceutical combination of any one of features 40-47, wherein the at least one RTI is administered at a daily dose that is less than or equal to about 80% of a standard adult monotherapy dose for treatment of viral infection, while maintaining a therapeutically effective level for modulation of age- or cancer-associated inflammation.

[0068] Feature 49: The pharmaceutical combination of any one of features 40-48, wherein component (a) is formulated for oral administration and component (b) is formulated for oral or parenteral administration.

[0069] Feature 50: A kit comprising (i) a first container containing a compound of formula (I) as defined in feature 1 or a pharmaceutical composition according to any one of features 28-38, (ii) a second container containing at least one reverse transcriptase inhibitor as defined in any one of features 41-43, and (iii) printed instructions for administration of the contents of the first and second containers as combination therapy to treat a disease as defined in feature 1.

[0070] Feature 51 : A method of selectively killing cancer cells in vitro, the method comprising contacting a population of cancer cells with an effective amount of a compound of formula (I) as defined in feature 1 under conditions sufficient to reverse an abnormal intracellular-to-extracellular pH gradient in the cancer cells and thereby induce apoptosis in the cancer cells.

[0071] Feature 52: The method of feature 51 , wherein the cancer cells comprise breast cancer cells, optionally estrogen receptor-positive MCF-7 cells or MCF-7-like cells and are cultured as monolayers or as multicellular spheroids.

[0072] Feature 53: The method of feature 51 or 52, further comprising measuring intracellular and / or extracellular pH before and after contacting the cells with the compound of formula (I) using a pH-sensitive fluorescent indicator.

[0073] Feature 54: A method for assessing the sensitivity of tumor cells from a subject to a compound of formula (I), the method comprising: (a) obtaining tumor cells from the subject; (b) contacting the tumor cells ex vivo with at least one test concentration of a compound of formula (I) as defined in feature 1; and (c) determining at least one response selected from a change in cell viability, a change in apoptosis and a change in intracellular or extracellular pH; wherein the magnitude of the response in step (c) is used to assess sensitivity of the tumor cells to the compound of formula (I).

[0074] Feature 55: The method of feature 54, further comprising, on the basis of the assessed sensitivity, selecting a treatment regimen for the subject that comprises administration of a therapeutically effective amount of a compound of formula (I).Docket No.: 411037-501001 WO

[0075] Feature 56: The method of any one of features 1-16, wherein the diseased cells exhibit an extracellular pH prior to treatment in the range of about 6.0 to about 7.0 and treatment with the compound of formula (I) increases the extracellular pH in the diseased tissue to a value in the range of about 7.0 to about 7.5.

[0076] Feature 57: A kit for assessing sensitivity of cancer cells to a compound of formula (I), the kit comprising: (a) a compound of formula (I) as defined in feature 1 ; (b) a pH-sensitive fluorescent indicator suitable for measuring intracellular and / or extracellular pH in living cells; and (c) printed instructions describing a protocol for contacting cancer cells with the compound of formula (I), detecting pH changes using the pH-sensitive fluorescent indicator, and determining sensitivity of the cancer cells to the compound of formula (I) based on the detected changes.

[0077] It is contemplated that the methods, compositions, formulations, and kits can be remote-operated, stable, easily boxed, shipped and portable in kits and / or in long-storage containers. It is important to note in the context of this disclosure that the materials and the methods can be used for production of kits, methods for assembling the kits, or for other constructs.

[0078] According to some aspects, the features, options, parameters, dimensions, materials, mounting configurations, arrangements, components, embodiments, and examples described in this Summary are each disclosed individually and in any technically sensible combination with one another and with features described elsewhere herein. Other implementations are also described and recited herein. These and other features and advantages will be apparent from a reading of the following detailed description and a review of the associated drawings. It is to be understood that both the foregoing general description and the following detailed description are explanatory only and are not restrictive of aspects as claimed.BRIEF DESCRIPTION OF THE DRAWINGS

[0079] For the purpose of illustration, certain embodiments of the present invention are shown in the drawings described below. It should be understood, however, that the invention is not limited to the precise arrangements, dimensions, and instruments shown. In the drawings:

[0080] FIG. 1A shows a plot of concentration versus absorbance for a cell viability assessment. FIG. 1B shows a plot of concentration versus absorbance for a cell viability assessment.

[0081] FIG. 2A shows images from the measurements of intracellular pH using pH Rodo indicators. FIG. 2B shows a plot of Chemical Target 4 concentration (pg / mL) versus mean fluorescence intensity.Docket No.: 411037-501001 WO

[0082] FIG. 3 shows images from results from the CellEvent™ Caspase-3 / 7 Assay.

[0083] FIG. 4 shows images from the pH Rodo Assay.

[0084] FIG. 5A shows a side-by-side comparison of images from the CellEvent™ Caspase-3 / 7 Assay and from the pH Rodo Assay. For the CellEvent™ Caspase-3 / 7 Assay, FIG. 5B shows a plot of (X-axis) the control and treated MCF-7 cells versus (Y-axis) mean fluorescence intensity (200 pm, see bottom of FIG. 5A). For the pH Rodo Assay, FIG. 5C shows a plot of (X-axis) the control and treated MCF-7 cells versus (Y-axis) mean fluorescence intensity (200 pm, see bottom of FIG. 5A).

[0085] FIG. 6A shows a conceptual overview of the 3-D MCF-7 spheroid model and the treatment outcome for MCF-7 breast adenocarcinoma cells. FIG. 6B shows the actual microscopy data.

[0086] FIG. 7 shows some IIIPAC (International Union of Pure and Applied Chemistry) names for some non-limiting example compounds.

[0087] FIG. 8 shows an example 13C NMR with chemical structure depicted at left.

[0088] FIG. 9 shows an example NMR spectrum with chemical structures depicted at left.

[0089] FIG. 10 shows chromatograms for the chemical compound shown at left in FIG. 9, and FIG. 10 also shows (at bottom) the integration results for the ELSD chromatogram. Diode array (DAD) signals of 220 nm and 254 nm are indicated on the chromatograms.

[0090] FIG. 11A shows the MS spectra at 0.311 minutes {e.g., see legend at top of FIG. 11 A) with the zoomed MS spectrum shown in FIG. 11B.

[0091] It should be understood that while different depictions, plots and / or colors with images could be sometimes used in some of the figures above to describe different embodiments and different aspects of the technology, any aspect from any figure can be inter-combined with any aspect from any other figures.DETAILED DESCRIPTION OF THE INVENTION

[0092] The subject innovation is now described in some instances with reference to the drawings. In the following description, for purposes of explanation, numerous specific details are set forth in order to provide a thorough understanding of the present invention. It may be evident, however, that the present invention may be practiced without these specific details. It is to be appreciated that certain aspects, modes, embodiments, variations and features of the invention are described below in various levels of detail in order to provide a substantial understanding of the present invention.Docket No.: 411037-501001 WODEFINITIONS

[0093] For convenience, the meaning of some terms and phrases used in the specification, examples, and appended claims, are provided below. Unless stated otherwise, or implicit from context, the following terms and phrases include the meanings provided below. The definitions are provided to aid in describing particular embodiments, and are not intended to limit the claimed invention, because the scope of the invention is limited only by the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is an apparent discrepancy between the usage of a term in the art and its definition provided herein, the definition provided within the specification shall prevail.

[0094] As used in this specification and the appended claims, the singular forms "a," "an" and "the" include plural referents unless the content clearly dictates otherwise. For example, reference to "a cell" includes a combination of two or more cells, and the like.

[0095] As used herein, the term "approximately" or "about" in reference to a value or parameter are generally taken to include numbers that fall within a range of 5%, 10%, 15%, or 20% in either direction (greater than or less than) of the number unless otherwise stated or otherwise evident from the context (except where such number would be less than 0% or exceed 100% of a possible value). As used herein, reference to "approximately" or "about" a value or parameter includes (and describes) embodiments that are directed to that value or parameter. For example, description referring to "about X" includes description of "X".

[0096] It is important to note that as used herein, numerical ranges and expressions of degree, such as “from X to Y,” “between X and Y,” “X-Y,” “at least X,” “no more than Y,” and the like, whether or not modified by terms such as “about” or “approximately,” are intended to encompass the stated endpoints as well as all intermediate values and all subranges included within the stated range, unless the context clearly indicates otherwise. Thus, for example, a range of from about 0.01 to about 100 includes values of 0.01, 0.02, 0.03, and so on, as well as 1, 2, 3, 4, and so forth up to and including 100, together with all sub-ranges such as from about 0.1 to about 10, from about 1 to about 50, or from about 5 to about 25, and the like. In all such instances, each individual value and each sub-range within the stated range is deemed to be expressly disclosed as if individually recited.

[0097] In the context of substituent and chain-length definitions, such as Ci-C4alkyl, C2-C6alkenyl, and C2-C6alkynyl, each permissible carbon chain length and each corresponding sub-range within the stated range is likewise intended to be expressly disclosed as if individually recited. Thus, by way of non-limiting example, the term “Ci-Docket No.: 411037-501001 WOC4alkyl” encompasses methyl (Ci), ethyl (C2), n-propyl and isopropyl (C3), and n-butyl, secbutyl, isobutyl and tert-butyl (C4), and further includes sub-ranges such as Ci-C3alkyl, C2- C4alkyl, or C2-C3alkyl. Similarly, the terms “C2-C6alkenyl” and “C2-C6alkynyl” encompass each chain length and position of unsaturation within the recited carbon range, together with all sub-ranges (for example C2-C4alkenyl, C3-C6alkenyl, C2-C5alkynyl, or C3-C4alkynyl), and each such value and sub-range is intended to be encompassed by the present disclosure as an individually supported embodiment.

[0098] As used herein, the term “or” means “and / or.” The term "and / or" as used in a phrase such as "A and / or B" herein is intended to include both A and B; A or B; A (alone); and B (alone). Likewise, the term "and / or" as used in a phrase such as "A, B, and / or C" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C;A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0099] As used herein, the term "comprising" means that other elements can also be present in addition to the defined elements presented. The use of "comprising" indicates inclusion rather than limitation.

[0100] The term "consisting of" refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the embodiment.

[0101] As used herein the term "consisting essentially of" refers to those elements required for a given embodiment. The term permits the presence of additional elements that do not materially affect the basic and novel or functional character! stic(s) of that embodiment of the invention.

[0102] As used herein, the term "statistically significant" or "significantly" refers to statistical significance and generally means a difference of at least two standard deviations (2 SD) from the mean, or a probability value (p-value) of less than 0.05 as determined by an appropriate statistical test, unless otherwise indicated.

[0103] As used herein, the term "subject" refers to a mammal, including but not limited to a dog, cat, horse, cow, pig, sheep, goat, chicken, rodent, or primate. Subjects can be house pets e.g., dogs, cats), agricultural stock animals {e.g., cows, horses, pigs, chickens, etc.), laboratory animals {e.g., mice, rats, rabbits, etc.), but are not so limited. Subjects include human subjects. The human subject may be a pediatric, adult, or a geriatric subject. The human subject may be of either sex.

[0104] As used herein, the terms "effective amount" and “therapeutically-effective amount” include an amount sufficient to prevent or ameliorate a manifestation of disease or medical condition, such as cancer. It will be appreciated that there will be many ways known in the art to determine the effective amount for a given application. For example, the pharmacological methods for dosage determination may be used in the therapeutic context.Docket No.: 411037-501001 WOIn the context of therapeutic or prophylactic applications, the amount of a composition administered to the subject will depend on the type and severity of the disease and on the characteristics of the individual, such as general health, age, sex, body weight and tolerance to drugs. It will also depend on the degree, severity and type of disease. The skilled artisan will be able to determine appropriate dosages depending on these and other factors. The compositions can also be administered in combination with one or more additional therapeutic compounds.

[0105] As used herein, the terms “treat,” “treatment,” “treating,” or “amelioration” when used in reference to a disease, disorder or medical condition, refer to therapeutic treatments for a condition, wherein the object is to reverse, alleviate, ameliorate, inhibit, slow down or stop the progression or severity of a symptom or condition. The term “treating” includes reducing or alleviating at least one adverse effect or symptom of a condition.Treatment is generally “effective” if one or more symptoms or clinical markers are reduced. Alternatively, treatment is “effective” if the progression of a condition is reduced or halted. That is, “treatment” includes not just the improvement of symptoms or markers, but also a cessation or at least slowing of progress or worsening of symptoms that would be expected in the absence of treatment. Beneficial or desired clinical results include, but are not limited to, alleviation of one or more symptom(s), diminishment of extent of the deficit, stabilized ( / .e., not worsening) state of a tumor or malignancy, delay or slowing of tumor growth and / or metastasis, and an increased lifespan as compared to that expected in the absence of treatment.

[0106] As used herein, the term "long-term" administration means that the therapeutic agent or drug is administered for a period of at least 12 weeks. This includes that the therapeutic agent or drug is administered such that it is effective over, or for, a period of at least 12 weeks and does not necessarily imply that the administration itself takes place for 12 weeks, e.g., if sustained release compositions or long-acting therapeutic agent or drug is used. Thus, the subject is treated for a period of at least 12 weeks. In many cases, long-term administration is for at least 4, 5, 6, 7, 8, 9 months or more, or for at least 1, 2, 3, 5, 7 or 10 years, or more.

[0107] The administration of the compositions contemplated herein may be carried out in any convenient manner, including by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation. In some embodiments, compositions are administered parenterally. The phrases “parenteral administration” and “administered parenterally” as used herein refers to modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravascular, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intratumoral, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular,Docket No.: 411037-501001 WO intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion. In one embodiment, the compositions contemplated herein are administered to a subject by direct injection into a tumor, lymph node, or site of infection.

[0108] The terms “decrease”, “reduced”, “reduction”, or “inhibit” are all used herein to mean a decrease by a statistically significant amount. In some embodiments, “reduce,” “reduction,” “decrease,” or “inhibit” typically means a decrease by at least 10% as compared to a reference level (e.g., the absence of a given treatment or agent) and can include, for example, a decrease by at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or more. As used herein, “reduction” or “inhibition” does not encompass a complete inhibition or reduction as compared to a reference level. “Complete inhibition” is a 100% inhibition as compared to a reference level. A decrease can preferably be down to a level accepted as within the range of normal for an individual without a given disorder.

[0109] The terms “increased”, “increase”, “enhance”, or “activate” are all used herein to mean an increase by a statistically significant amount. In some embodiments, the terms “increased”, “increase”, “enhance”, or “activate” can mean an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level. In the context of a marker or symptom, an “increase” is a statistically significant increase in such level.Cancer-related definitions:

[0110] As used herein, the term “cancer” relates generally to a class of diseases or conditions in which abnormal cells divide without control and can invade nearby tissues. Cancer cells can also spread to other parts of the body through the blood and lymph systems. There are several main types of cancer. Carcinoma is a cancer that begins in the skin or in tissues that line or cover internal organs. Sarcoma is a cancer that begins in bone, cartilage, fat, muscle, blood vessels, or other connective or supportive tissue. Leukemia is a cancer that starts in blood-forming tissue such as the bone marrow and causes large numbers of abnormal blood cells to be produced and enter the blood. Lymphoma andDocket No.: 411037-501001 WO multiple myeloma are cancers that begin in the cells of the immune system. Central nervous system cancers are cancers that begin in the tissues of the brain and spinal cord.

[0111] In some embodiments of any of the aspects, the cancer is a primary cancer. In some embodiments of any of the aspects, the cancer is a malignant cancer. As used herein, the term “malignant” refers to a cancer in which a group of tumor cells display one or more of uncontrolled growth ( / .e., division beyond normal limits), invasion ( / .e., intrusion on and destruction of adjacent tissues), and metastasis ( / .e., spread to other locations in the body via lymph or blood). As used herein, the term “metastasize” refers to the spread of cancer from one part of the body to another. A tumor formed by cells that have spread is called a “metastatic tumor” or a “metastasis.” The metastatic tumor contains cells that are like those in the original (primary) tumor.

[0112] As used herein, the term “benign” or “non-malignant” refers to tumors that may grow larger but do not spread to other parts of the body. Benign tumors are self-limited and typically do not invade or metastasize.

[0113] A “tumor” refers generally to a swelling or lesion formed by an abnormal growth of cells, which may be benign, pre-malignant, or malignant. Most cancer cells form tumors, but some, for example leukemia cells, do not necessarily form tumors. For those cancers that form tumors, the terms “cancer cell” and “tumor cell” can be used interchangeably.

[0114] A subject that has a cancer or a tumor is a subject having objectively measurable cancer cells present in the subject’s body. Included in this definition are malignant, actively proliferative cancers, as well as potentially dormant tumors or micrometastases. Cancers which migrate from their original location and seed other vital organs can eventually lead to the death of the subject through the functional deterioration of the affected organs. Hematopoietic cancers, such as leukemia, are able to out-compete the normal hematopoietic compartments in a subject, thereby leading to hematopoietic failure (in the form of anemia, thrombocytopenia and neutropenia) ultimately causing death.

[0115] Examples of cancer include but are not limited to carcinoma, lymphoma, blastoma, sarcoma, leukemia, basal cell carcinoma, biliary tract cancer, bladder cancer, bone cancer, brain and central nervous system (CNS) cancer, breast cancer, cancer of the peritoneum, cervical cancer, choriocarcinoma, colon and rectum cancer, connective tissue cancer, cancer of the digestive system, endometrial cancer, esophageal cancer, eye cancer, cancer of the head and neck, gastric cancer (including gastrointestinal cancer), glioblastoma (GBM), hepatic carcinoma, hepatoma, intra-epithelial neoplasm, kidney or renal cancer, larynx cancer, liver cancer, lung cancer (e.g., small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung), melanoma, myeloma, neuroblastoma, oral cavity cancer (e.g., lip, tongue, mouth, and pharynx), ovarianDocket No.: 411037-501001 WO cancer, pancreatic cancer, prostate cancer, retinoblastoma, rhabdomyosarcoma, rectal cancer, cancer of the respiratory system, salivary gland carcinoma, skin cancer, squamous cell cancer, stomach cancer, testicular cancer, thyroid cancer, uterine or endometrial cancer, cancer of the urinary system, vulval cancer, as well as other carcinomas and sarcomas; as well as B-cell lymphoma (including low grade / follicular non-Hodgkin’s lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade / follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small noncleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom’s Macroglobulinemia); chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); hairy cell leukemia; chronic myeloblastic leukemia; and posttransplant lymphoproliferative disorder (PTLD); as well as abnormal vascular proliferation associated with phakomatoses, edema (such as that associated with brain tumors), and Meigs’ syndrome.

[0116] A “cancer cell” is a cancerous, pre-cancerous, or transformed cell, either in vivo, ex vivo, or in tissue culture, that has spontaneous or induced phenotypic changes that do not necessarily involve the uptake of new genetic material. Although transformation can arise from infection with a transforming virus and incorporation of new genomic nucleic acid, or uptake of exogenous nucleic acid, it can also arise spontaneously or following exposure to a carcinogen, thereby mutating an endogenous gene. Transformation and / or cancer is associated with, for example, morphological changes, immortalization of cells, aberrant growth control, foci formation, anchorage independence, malignancy, loss of contact inhibition and density limitation of growth, growth factor or serum independence, tumorspecific markers, invasiveness or metastasis, and tumor growth in suitable animal hosts such as nude mice.

[0117] A subject can be one who has been previously diagnosed with or identified as suffering from or having a condition in need of treatment (e.g., a cancer) or one or more complications related to such a condition, and optionally, but need not have already undergone treatment for a condition or the one or more complications related to the condition. Alternatively, a subject can also be one who has not been previously diagnosed as having a condition in need of treatment or one or more complications related to such a condition. For example, a subject can be one who exhibits one or more risk factors for a condition or one or more complications related to a condition or a subject who does not exhibit risk factors. A “subject in need” of treatment for a particular condition can be a subject having that condition, diagnosed as having that condition, or at risk of developing that condition.Docket No.: 411037-501001 WOPHARMACEUTICAL COMPOSITIONS

[0118] The compositions and methods of the present invention may be utilized to treat an individual in need thereof. In certain embodiments, the individual is a mammal such as a human, or a non-human mammal. When administered to an animal, such as a human, the composition or the compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. In some embodiments, when such pharmaceutical compositions are for human administration, particularly for invasive routes of administration ( / .e., routes, such as injection or implantation, that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free, or substantially pyrogen-free. The excipients can be chosen, for example, to effect delayed release of an agent or to selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form such as tablet, capsule (including sprinkle capsule and gelatin capsule), granule, lyophile for reconstitution, powder, solution, syrup, suppository, injection or the like. The composition can also be present in a transdermal delivery system, e.g., a skin patch. The composition can also be present in a solution suitable for topical administration, such as a lotion, cream, or ointment.

[0119] A pharmaceutically acceptable carrier can contain physiologically acceptable agents that act, for example, to stabilize, increase solubility or to increase the absorption of a compound such as a compound of the invention. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition can be a self-emulsifying drug delivery system or a self-micro emulsifying drug delivery system. The pharmaceutical composition (preparation) also can be a liposome or other polymer matrix, which can have incorporated therein, for example, a compound of the invention. Liposomes, for example, which comprise phospholipids or other lipids, are nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer.

[0120] The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings andDocket No.: 411037-501001 WO animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0121] The phrase "pharmaceutically acceptable carrier" as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

[0122] A pharmaceutical composition (preparation) can be administered to a subject by any of a number of routes of administration including, for example, orally (for example, drenches as in aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); absorption through the oral mucosa (e.g., sublingually); subcutaneously; transdermally (for example as a patch applied to the skin); and topically (for example, as a cream, ointment or spray applied to the skin). The compound may also be formulated for inhalation. In certain embodiments, a compound may be simply dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions suitable for same can be found in, for example, U.S. Patent Nos. 6,110,973, 5,763,493, 5,731 ,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, as well as in patents cited therein.

[0123] The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent toDocket No.: 411037-501001 WO about ninety-nine percent of active ingredient, preferably from about 5 percent to about 70 percent, most preferably from about 10 percent to about 30 percent.

[0124] Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the invention, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0125] Formulations of the invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), lyophile, powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and / or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. Compositions or compounds may also be administered as a bolus, electuary or paste.

[0126] To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complexing agents, such as modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.Docket No.: 411037-501001 WO

[0127] A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropyl methyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0128] The tablets, and other solid dosage forms of the pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropyl methyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

[0129] Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophiles for reconstitution, micro-emulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

[0130] Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

[0131] Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol andDocket No.: 411037-501001 WO sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

[0132] Dosage forms for the topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be required.

[0133] The ointments, pastes, creams and gels may contain, in addition to an active compound, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0134] Powders and sprays can contain, in addition to an active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

[0135] Transdermal patches have the added advantage of providing controlled delivery of a compound of the present invention to the body. Such dosage forms can be made by dissolving or dispersing the active compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

[0136] The phrases "parenteral administration" and "administered parenterally" as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intraocular (such as intravitreal), intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

[0137] Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixturesDocket No.: 411037-501001 WO thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0138] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

[0139] In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

[0140] Injectable depot forms are made by forming microencapsulated matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissue.

[0141] For use in the methods of this invention, active compounds can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically- acceptable carrier.

[0142] Methods of introduction may also be provided by rechargeable or biodegradable devices. Various slow release polymeric devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including proteinaceous biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form an implant for the sustained release of a compound at a particular target site.Docket No.: 411037-501001 WO

[0143] Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

[0144] The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound(s) being employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound(s) employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0145] A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the pharmaceutical composition or compound at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. By “therapeutically effective amount” is meant the concentration of a compound that is sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the weight, sex, age, and medical history of the subject. Other factors which influence the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being administered with the compound of the invention. A larger total dose can be delivered by multiple administrations of the agent. Methods to determine efficacy and dosage are known to those skilled in the art. See, e.g., Isselbacher et al. (1996).1

[0146] In general, a suitable daily dose of an active compound used in the compositions and methods of the invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

[0147] If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. In certain embodiments of the present invention, the active compound may be administered two or three times daily. In other embodiments, the active compound will be administered once daily.Docket No.: 411037-501001 WO

[0148] The patient receiving this treatment is any animal in need, including primates, in particular humans; and other mammals such as equines bovine, porcine, sheep, feline, and canine; poultry; and pets in general.

[0149] In certain embodiments, compounds of the invention may be used alone or conjointly administered with another type of therapeutic agent.

[0150] The present disclosure includes the use of pharmaceutically acceptable salts of compounds of the invention in the compositions and methods of the present invention. In certain embodiments, contemplated salts of the invention include, but are not limited to, alkyl, dialkyl, trialkyl or tetra-alkyl ammonium salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, L-arginine, benenthamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2- (diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, 1-hydroxy-2-naphthoic acid, 2,2- dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, l-ascorbic acid, l-aspartic acid, benzenesulfonic acid, benzoic acid, (+)-camphoric acid, (+)-camphor-10-sulfonic acid, capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, d-glucoheptonic acid, d-gluconic acid, d-glucuronic acid, glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, l-malic acid, malonic acid, mandelic acid, methanesulfonic acid , naphthalene-1,5-disulfonic acid, naphthalene-2- sulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, proprionic acid, l-pyroglutamic acid, salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, l-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, and undecylenic acid salts.

[0151] The pharmaceutically acceptable acid addition salts can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent.Docket No.: 411037-501001 WO

[0152] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0153] Examples of pharmaceutically acceptable antioxidants include: (1) water- soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0154] Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art to which this disclosure belongs. It should be understood that this invention is not limited to the particular methodology, protocols, and reagents, etc., described herein and as such can vary. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention, which is defined solely by the claims. Definitions of common terms in immunology and molecular biology can be found in The Merck Manual of Diagnosis and Therapy;2The Encyclopedia of Molecular Cell Biology and Molecular Medicine;3Molecular Biology and Biotechnology: a Comprehensive Desk Reference;4Immunology;5Janeway's Immunobiology;6Lewin's Genes XI;7Molecular Cloning: A Laboratory Manual.;8Basic Methods in Molecular Biology;9Laboratory Methods in Enzymology;10Current Protocols in Molecular Biology (CPMB);11Current Protocols in Protein Science (CPPS);12and Current Protocols in Immunology (CPI).13

[0155] In some embodiments of any of the aspects, the disclosure described herein does not concern a process for cloning human beings, processes for modifying the germ line genetic identity of human beings, uses of human embryos for industrial or commercial purposes or processes for modifying the genetic identity of animals which are likely to cause them suffering without any substantial medical benefit to man or animal, and also animals resulting from such processes.

[0156] Other terms are defined herein within the description of the various aspects of the invention.CANCER HOMEOSTASIS DISRUPTION TECHNOLOGY

[0157] In some embodiments, the present invention provides a novel class of glycosidase-resistant glucose C-glycosyl basic amino acid conjugates of formula (I), G-L- NH-CO-AA, that are configured to disrupt cancer homeostasis by controlled modulation ofDocket No.: 411037-501001 WO intracellular and extracellular pH gradients. According to some aspects, the glucose C- glycosyl residue G acts as a recognition motif for glucose transport systems upregulated in malignant and metabolically stressed cells, whereas the linker segment L and the basic amino acid residue AA cooperate to provide a pH-responsive scaffold capable of binding, redistributing, or buffering protons within a tumor microenvironment. In some embodiments, this cooperative behavior produces a net reversal or normalization of the abnormal intracellular-to-extracellular pH gradient that is characteristic of many cancers, thereby promoting apoptosis and loss of long-term clonogenic survival in diseased cells.

[0158] According to some aspects, compounds of formula (I) differ from classical iontransport or proton-pump inhibitors by exploiting a glycosidase-resistant carbon-carbon bond between the anomeric carbon (C-1) of the glucose C-glycosyl residue and the proximal carbon atom of the linker L. This C-glycosidic architecture confers resistance to enzymatic cleavage by glycosidases and related hydrolases and thus supports sustained exposure of tumor cells to the intact G-L-NH-CO-AA framework in both extracellular and intracellular compartments. In some embodiments, the linker L is selected to modulate spatial orientation, conformational flexibility, and overall lipophilicity of the conjugate, while the basic amino acid residue AA provides a side chain with a pKa suitable for protonation within acidic extracellular regions and deprotonation within relatively alkaline intracellular spaces. In a non-limiting example, AA can be a lysine- or arginine-derived residue that contributes additional cationic centers for proton capture and facilitates interaction with negatively charged cellular membranes.

[0159] In some embodiments, the functional performance of these G-L-NH-CO-AA conjugates has been evaluated in vitro using human breast adenocarcinoma MCF-7 cells as a representative solid-tumor model. FIG. 1A and FIG. 1B illustrate non-limiting examples of cell-viability data obtained with a prototypical compound of formula (I), referred to herein as Chemical Target 4, in a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay after a 48-hour incubation period. In FIG. 1A, exposure of MCF-7 cells to gradually increasing concentrations of Chemical Target 4 over a lower dose range (for example 0-200 pg / mL) produces a concentration-dependent decrease in absorbance at 570 nm, after correction for background at 630 nm, consistent with reduced mitochondrial metabolic activity and diminished viability relative to vehicle-treated controls. In FIG. 1B, extension of the concentration range to higher levels (for example up to 1000 pg / mL) demonstrates a further progressive loss of viability and highlights the capacity of the G-L- NH-CO-AA scaffold to achieve robust cytotoxic effects within this model.

[0160] According to some aspects, the data in FIG. 1A and FIG. 1B provide evidence that compounds of formula (I) exert cytostatic and cytotoxic actions in cancer cells at sub-millimolar to low-millimolar concentrations, with a sigmoidal dose-response profileDocket No.: 411037-501001 WO compatible with a defined molecular mechanism rather than nonspecific detergent-like lysis. In some embodiments, the observed decrease in MTT signal at intermediate concentrations corresponds to an IC50or IC30range that can be used for further mechanistic studies, including pH-sensitive imaging and apoptosis assays described in subsequent figures. In this example, Chemical Target 4 serves as an exemplar of the broader class of glucose C- glycosyl basic amino acid conjugates and demonstrates that the G-L-NH-CO-AA architecture is compatible with cellular uptake, intracellular engagement, and biologically meaningful modulation of cancer cell viability.

[0161] In a non-limiting example, the viability curves in FIG. 1A and FIG. 1B are interpreted in conjunction with pH-responsive fluorescence measurements and caspase-3 / 7 activation readouts to support a model in which compounds of formula (I) perturb both intracellular and pericellular proton distribution. According to some aspects, uptake of a compound of formula (I) through glucose transport pathways enriches the conjugate within cancer cells that exhibit elevated glycolytic flux and a reversed pH gradient. Subsequent protonation-deprotonation cycling of the AA side chain and associated buffering effects at cellular membranes are believed to drive acidification of the intracellular compartment and partial neutralization of acidic extracellular pockets. In some embodiments, this reestablishment of a more physiological pH gradient destabilizes pro-survival signaling networks, promotes activation of executioner caspases, and sensitizes tumor cells to additional stresses or combination therapies.

[0162] In some embodiments, FIG. 1A and FIG. 1B thus provide foundational quantitative support for the concept of “cancer homeostasis disruption technology” described in this specification. The reduction in MCF-7 cell viability observed upon treatment with a representative G-L-NH-CO-AA conjugate confirms that the glucose C-glycosyl residue, the glycosidase-resistant carbon-carbon linkage, and the basic amino acid moiety can be assembled into a compact small-molecule framework that produces potent effects on tumor cell survival. According to some aspects, these data justify further exploration of structureactivity relationships within the G, L and AA domains, optimization of dosing regimens and formulation strategies, and translation of the platform to additional cancer types, inflammatory indications, and chronic autoimmune disorders characterized by pathological pH gradients. In a non-limiting example, the same design principles can be applied to generate analogs with tailored pharmacokinetic properties, altered transporter selectivity, or combination-therapy compatibility, thereby extending the scope of cancer homeostasis disruption technology beyond the specific exemplar illustrated by Chemical Target 4 (discussed below).

[0163] In some embodiments, the next set of experimental results assesses whether compounds of formula (I), G-L-NH-CO-AA, alter intracellular pH homeostasis in cancer cellsDocket No.: 411037-501001 WO in a manner consistent with the cancer homeostasis disruption technology described herein. According to some aspects, MCF-7 breast adenocarcinoma cells are exposed for 24 hours to a range of concentrations of a representative compound of formula (I), designated Chemical Target 4, and intracellular pH is monitored using a pH-sensitive fluorogenic indicator of the pH rodo type. The pH rodo probe exhibits low fluorescence at near-neutral pH and progressively higher fluorescence as the local environment becomes more acidic, thereby providing a convenient readout of pH perturbation within endosomal, lysosomal, or pericellular acidic compartments.

[0164] In this example, FIG. 2A presents qualitative fluorescence micrographs obtained from the pH rodo assay at different treatment conditions. The panels in FIG. 2A show, from left to right, transmitted-light images of the culture wells, higher magnification views of MCF-7 cells, and corresponding red fluorescence images highlighting pH rodo signal intensity. Untreated control cells, which receive only vehicle (for example, dimethyl sulfoxide at concentrations matching those used to deliver Chemical Target 4), exhibit relatively low pH rodo fluorescence with only scattered punctate signals, consistent with basal levels of acidic organelles in metabolically active epithelial cells. In contrast, cultures treated with increasing concentrations of Chemical Target 4 display progressively brighter and more widespread red fluorescence, with numerous fluorescent puncta and regions of enhanced signal intensity distributed across the cell layer. According to some aspects, the visual comparison in FIG. 2A indicates that exposure to the G-L-NH-CO-AA scaffold leads to measurable changes in intracellular or pericellular acidity relative to vehicle controls.

[0165] In some embodiments, the qualitative observations from FIG. 2A are supported by quantitative analysis summarized in FIG. 2B. FIG. 2B shows a bar graph of mean fluorescence intensity plotted as a function of Chemical Target 4 concentration (e.g., 0, 50, 100, 200, 500, and 1000 pg / mL). Each bar represents the mean pH rodo signal measured across replicate wells, with error bars indicating the standard error of the mean. As the concentration of Chemical Target 4 increases, mean fluorescence intensity rises in a concentration-dependent manner, with statistically significant elevations at intermediate and higher doses relative to the untreated baseline, as indicated by asterisks above the bars. According to some aspects, the monotonic increase in mean fluorescence intensity demonstrates that compounds of formula (I) elicit graded perturbations of intracellular pH, rather than an all-or-nothing effect, and confirms that the observed changes are reproducible across independent replicates.

[0166] In a non-limiting example, the combined data from FIG. 2A and FIG. 2B are interpreted as evidence that the G-L-NH-CO-AA framework engages proton-handling processes within cancer cells and shifts the balance of acidic and neutral compartments. In some embodiments, increased pH rodo fluorescence reflects enhanced acidification ofDocket No.: 411037-501001 WO endosomal or lysosomal structures, or redistribution of protons toward compartments labeled by the indicator, in response to the presence of protonatable basic amino acid residues within the AA segment of formula (I). When considered together with the viability data in FIG. 1A and FIG. 1B, the pH rodo results support a model in which compounds of formula (I) disrupt the abnormal pH homeostasis of tumor cells, producing both biochemical changes in proton gradients and functional consequences for cell survival.

[0167] In some embodiments, three-dimensional tumor spheroid models are employed to evaluate cancer homeostasis disruption by compounds of formula (I), G-L-NH- CO-AA, under conditions that better approximate the architecture and diffusion barriers of solid tumors. According to some aspects, human MCF-7 breast cancer cells are seeded at 5,000 cells per well and allowed to aggregate over several days to form compact spheroids with smooth contours. FIG. 3 and FIG. 4 illustrate non-limiting examples of such spheroids at successive stages of aggregation and treatment. The panels in each figure are organized from left to right to show spheroid morphology on the day after plating (Day -1), spheroid appearance on Day 0, spheroid appearance on Day 2 after a 48-hour treatment period, and fluorescence micrographs obtained with functional probes on Day 2. Separate rows depict treated spheroids and untreated control spheroids, with a scale bar of 200 pm for reference.

[0168] In FIG. 3, the rightmost column presents images from a CellEvent™ Caspase-3 / 7 assay that reports activation of executioner caspases associated with apoptosis. Control spheroids exhibit dense, roughly spherical morphology with sharp boundaries and only sparse green fluorescence, consistent with low basal caspase-3 / 7 activity. In contrast, spheroids exposed to a representative compound of formula (I) display pronounced architectural disruption after 48 hours, including irregular outlines and partial disaggregation at the periphery. The corresponding caspase-3 / 7 images show numerous bright fluorescent foci distributed throughout the treated spheroids, indicating widespread activation of apoptotic pathways. According to some aspects, these observations demonstrate that compounds of formula (I) induce programmed cell death not only in monolayer cultures but also within the more complex microenvironment of multicellular spheroids.

[0169] FIG. 4 presents an analogous series of images obtained from a pH rodo- based assay under the same spheroid culture conditions. In this example, the rightmost column displays red fluorescence corresponding to regions of increased acidity. Untreated control spheroids show relatively modest pH rodo signal confined mainly to peripheral zones, whereas treated spheroids exhibit intensified and more extensive red fluorescence, particularly at regions of structural disruption. According to some aspects, the enhanced pH rodo signal in treated spheroids reflects substantial perturbation of intra-spheroidal andDocket No.: 411037-501001 WO pericellular pH, consistent with redistribution or sequestration of protons by the G-L-NH-CO- AA scaffold.

[0170] In combination, FIG. 3 and FIG. 4 provide visual evidence that compounds of formula (I) remodel spheroid morphology, increase caspase-3 / 7 activation, and alter acidic microdomains within three-dimensional tumor constructs. In some embodiments, these findings support a mechanistic model in which pH-gradient disruption within a constrained tumor-like geometry leads to loss of structural integrity, initiation of apoptosis, and progressive collapse of the malignant cell population.

[0171] In some embodiments, increased pH-sensitive fluorescence reflects redistribution and buffering of protons within tumor cells and adjacent microdomains, while the bulk extracellular pH is shifted toward physiological values. In this example, pH rodo is only reporting local pockets of lower pH (e.g., endo-lysosomal or pericellular domains), and the overall effect is a re-balancing of the intracellular / extracellular gradient, not a net increase of extracellular acidity.

[0172] In some embodiments, further experiments in three-dimensional MCF-7 spheroid cultures compare the activity of a representative compound of formula (I), G-L-NH- CO-AA, with a conventional chemotherapeutic agent. According to some aspects, FIG. 5A presents photomicrographs from both the pH rodo assay and the CellEvent™ caspase-3 / 7 assay under matched spheroid culture conditions. The upper row in FIG. 5A shows spheroids treated with a positive control concentration of doxorubicin (for example 10 nM), which is known to induce apoptosis and to disrupt tumor spheroid integrity. The phasecontrast images of doxorubicin-treated spheroids display irregular contours and partial fragmentation, while the corresponding red pH rodo fluorescence images reveal intense signal at multiple foci, consistent with pronounced acidification in discrete microdomains.

[0173] In the middle rows of FIG. 5A, spheroids exposed to Chemical Target 4, a non-limiting example of a compound of formula (I), exhibit morphological alterations and fluorescence patterns that parallel those observed with doxorubicin. Treated spheroids display less compact architecture and show bright pH rodo fluorescence in peripheral and internal regions, indicating substantial perturbation of acidic compartments. By contrast, control MCF-7 spheroids in the lower pH rodo panel retain smoother outlines and show only modest red fluorescence, reflecting basal acidity. According to some aspects, these observations support the conclusion that the G-L-NH-CO-AA scaffold elicits pH changes in three-dimensional tumor structures that are qualitatively comparable to those produced by a standard cytotoxic agent.

[0174] The lower half of FIG. 5A depicts the CellEvent™ caspase-3 / 7 assay for the same experimental series. Spheroids treated with Chemical Target 4 show robust green fluorescence distributed throughout the structure, indicative of widespread activation ofDocket No.: 411037-501001 WO executioner caspases and ongoing apoptosis. In contrast, control spheroids display only sparse green puncta, suggesting low basal caspase activity.

[0175] Quantitative analyses corresponding to these images are summarized in FIG. 5B and FIG. 5C. FIG. 5B presents mean fluorescence intensity values for the caspase- 3 / 7 assay, comparing treated and untreated MCF-7 spheroids. The bar graph demonstrates a statistically significant increase in caspase-3 / 7 signal in the presence of Chemical Target 4, as indicated by triple asterisks above the treated bar. FIG. 5C shows the analogous comparison for the pH rodo assay, where mean fluorescence intensity is markedly higher in treated spheroids than in controls, again with high statistical significance. According to some aspects, the combined results of FIGs. 5A-5C indicate that compounds of formula (I) not only remodel pH microdomains within tumor spheroids but also trigger apoptotic pathways to a degree comparable to or greater than established chemotherapeutic controls, thereby providing functional confirmation of cancer homeostasis disruption in a physiologically relevant three-dimensional context.

[0176] In some embodiments, FIG. 6A and FIG. 6B provide an integrated schematic and imaging-based summary of cancer homeostasis disruption technology as applied to three-dimensional tumor spheroids treated with a representative compound of formula (I), G- L-NH-CO-AA. According to some aspects, the left-hand schematic of FIG. 6A illustrates the typical evolution of MCF-7 spheroids from loose cellular aggregates on Day 1 , through tight aggregation on Day 2, to the formation of smooth-contoured, compact spheroids by Day 7. Upon subsequent exposure to a potential anti-cancer peptide or peptidomimetic of formula (I) at, for example, 100 pg / mL for 48 hours, the spheroids undergo a transition to irregular, size-reduced structures populated by a mixture of viable, dying and dead cancer cells, as depicted by the graded colour coding of individual cells in the schematic.

[0177] In this example, the panels on the right side of FIG. 6A provide corresponding experimental micrographs that visualize structural and functional changes in treated versus untreated spheroids. The upper pair of images for the pH rodo assay compare a treated spheroid and an untreated spheroid. The treated spheroid exhibits an irregular outline and fragmented architecture, accompanied by intense red pH rodo fluorescence that marks multiple acidic microdomains within and around the spheroid. The untreated spheroid maintains a smoother contour and displays only modest red fluorescence, indicative of basal levels of acidity. According to some aspects, this contrast supports the conclusion that administration of a compound of formula (I) reconfigures proton distribution within the spheroid microenvironment, leading to enhanced localization of acidic compartments in association with structural disintegration.

[0178] FIG. 6B presents an analogous comparison using the CellEvent™ caspase- 3 / 7 assay, which reports activation of executioner caspases associated with apoptosis. InDocket No.: 411037-501001 WO the treated condition, the spheroid appears morphologically disrupted and shows bright green fluorescence corresponding to active caspase-3 / 7, distributed across central and peripheral regions. In the untreated condition, the spheroid retains an intact, rounded morphology and exhibits only sparse green puncta, consistent with low basal apoptotic activity. In some embodiments, the combination of these images demonstrates that compounds of formula (I) not only modulate pH-related readouts but also trigger canonical apoptotic pathways in a three-dimensional tumor context.

[0179] According to some aspects, FIG. 6A and FIG. 6B together depict a coherent sequence in which spheroids formed from MCF-7 cells first establish a characteristic architecture and then, following treatment with a G-L-NH-CO-AA conjugate, undergo pH- gradient disruption, enhanced acidification in discrete regions, activation of caspase-3 / 7, and macroscopic collapse of spheroid structure. In a non-limiting example, these findings support a model wherein glucose C-glycosyl basic amino acid conjugates of formula (I) enter tumor spheroids preferentially, reprogram proton flux and buffering capacity, and drive the system away from a cancer-favoring homeostatic state toward either restoration of near- physiological conditions or apoptotic elimination of malignant cells.

[0180] A plurality of details (including detailed methods and conceptions) were written down and are now presented and contemplated and put forth as inter-changeable examples during this detailed description. In a discussion, study or a reading of the details, features, embodiments, aspects, any figure or any part of any figure, and / or examples of the technology disclosed herein, any of the features, embodiments, aspects, and / or examples herein can be optionally inter-combined (or inter-discussed) with the example details listed below, and any portion (or aspect) of any detail below can be inter-combined with any portion of any feature or example disclosed herein:

[0181] Detail 1: A compound of formula (I):G-L-NH-CO-AA (Formula I),(D-glucose),Docket No.: 411037-501001 WO(L-glucose),

[0182] wherein: G is an L-glucose C-glycosyl residue or a D-glucose C-glycosyl residue in the pyranose form having the carbon numbering shown in the figure of formula (I), said residue having a six-membered ring with a -CH2OH substituent at C-5 and hydroxyl groups at C-2, C-3, C-4 and C-6, wherein each of the hydroxyl groups at C-2, C-3, C-4 and C-6 is independently present as an unsubstituted hydroxyl group or as a group OR7, SR7, NR8R9, O-C(=O)R10or O-C(=O)OR10, and wherein the anomeric carbon (C-1) is bonded through a carbon-carbon bond B1 to a carbon atom of the linker group L; R7is hydrogen, Ci- C4alkyl, C2-C6alkenyl, C2-C6alkynyl, benzyl, phenyl or phenyl substituted by one or more Ci-C4alkyl, C1-C4 alkoxy, halogen, hydroxy or nitro; R8and R9are independently hydrogen or C1-C4 alkyl; and R10is Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, benzyl or phenyl; L is a divalent linker group connecting the anomeric carbon (C-1) of G to the nitrogen atom of the - NH-CO- group and is selected from Ci-C6straight or branched alkylene and C2-C6alkylene optionally interrupted by one or more heteroatoms selected from O, S and NRa, wherein L contains from 1 to 8 carbon atoms in total and Rais hydrogen or Ci-C4alkyl; AA is the residue of a basic amino acid of formula -NH-CHR11-(CH2)P-NR2R3-CO-, which is amide- bonded through its a-carboxyl group to the -NH-CO- moiety of formula (I), wherein: p is an integer from 2 to 5; R11is hydrogen, Ci-C6alkyl optionally substituted by hydroxy, amino, carboxy, Ci-C4alkoxycarbonyl or halogen, or is -(CH2)m-NR4R5where m is 1-4 and R4and R5are independently hydrogen or Ci-C4alkyl; R2and R3are independently hydrogen, Ci-C4alkyl, hydroxy-Ci-C4alkyl, or a group -C(=NH)NH2, or R2and R3together with the nitrogen atom to which they are attached form a saturated 5-, 6- or 7-membered heterocycle optionally containing an additional nitrogen atom; with the proviso that AA is a residue of a basic amino acid selected from: lysine, arginine, ornithine, homoarginine, histidine, diaminobutyric acid or diaminopropionic acid, or a non-naturally occurring analog thereof having a side chain pKa of at least 7; and including any enantiomer, diastereomer, racemateDocket No.: 411037-501001 WO or mixture thereof, and any pharmaceutically acceptable salt, solvate, hydrate or prodrug of a compound of formula (I).

[0183] Detail 2: The compound of detail 1, wherein G is an a-D-glucopyranosyl C- glycosyl residue.

[0184] Detail 3: The compound of detail 1, wherein G is a p-D-glucopyranosyl C- glycosyl residue.

[0185] Detail 4: The compound of detail 1, wherein G is an a-L-glucopyranosyl C- glycosyl residue.

[0186] Detail 5: The compound of detail 1, wherein G is a p-L-glucopyranosyl C- glycosyl residue.

[0187] Detail 6: The compound of any one of details 1-5, wherein AA is in the L- configuration at the a-carbon.

[0188] Detail 7: The compound of detail 6, wherein AA is the residue of lysine.

[0189] Detail 8: The compound of detail 6, wherein AA is the residue of arginine.

[0190] Detail 9: The compound of any one of details 1-8, further comprising another therapeutic agent selected from: (i) a cytotoxic chemotherapeutic agent; (ii) a targeted smallmolecule kinase inhibitor; (iii) an immune checkpoint inhibitor; (iv) a PARP inhibitor; (v) a hormone or endocrine therapy agent; (vi) a monoclonal antibody or antibody-drug conjugate; (vii) an epigenetic agent and / or modulator; and (viii) a cell-based immunotherapy.

[0191] Detail 10: The compound of detail 9, wherein the additional therapeutic agent is a cytotoxic chemotherapeutic agent selected from: Cyclophosphamide, Ifosfamide, Melphalan, Chlorambucil, Busulfan, Dacarbazine, Temozolomide, Cisplatin, Carboplatin, Oxaliplatin, Doxorubicin, liposomal Doxorubicin, Epirubicin, Daunorubicin, Idarubicin, Mitoxantrone, 5-Fluorouracil (5-Fll), Capecitabine, Gemcitabine, Cytarabine, Methotrexate, Pemetrexed, Paclitaxel, Docetaxel, Nab-paclitaxel (albumin-bound paclitaxel), Vincristine, Vinblastine, Vinorelbine, Eribulin, Irinotecan, Topotecan, Etoposide, Bleomycin, and Mitomycin C.

[0192] Detail 11: The compound of detail 9, wherein the additional therapeutic agent is a targeted small-molecule kinase inhibitor selected from: Imatinib, Dasatinib, Nilotinib, Bosutinib, Ponatinib, Erlotinib, Gefitinib, Afatinib, Osimertinib, Crizotinib, Ceritinib, Alectinib, Brigatinib, Lorlatinib, Sunitinib, Sorafenib, Pazopanib, Axitinib, Regorafenib, Cabozantinib, Lenvatinib, Vemurafenib, Dabrafenib, Encorafenib, Trametinib, Cobimetinib, Binimetinib, Palbociclib, Ribociclib, Abemaciclib, Everolimus, Temsirolimus, Alpelisib, Copanlisib, Idelalisib, Ibrutinib, Acalabrutinib, and Zanubrutinib.

[0193] Detail 12: The compound of detail 9, wherein the additional therapeutic agent is an immune checkpoint inhibitor selected from: an Anti-PD-1 antibody, Nivolumab, Pembrolizumab, Cemiplimab, Dostarlimab, Retifanlimab, Tislelizumab, Toripalimab,Docket No.: 411037-501001 WOPenpulimab, an Anti-PD-L1 antibody, Atezolizumab, Durvalumab, Avelumab, Cosibelimab, an Anti-CTLA-4 antibody, Ipilimumab, Tremelimumab, an Anti-l_AG-3 antibody, and Relatlimab.

[0194] Detail 13: The compound of detail 9, wherein the additional therapeutic agent is a PARP inhibitor selected from: Olaparib, Niraparib, Rucaparib, and Talazoparib.

[0195] Detail 14: The compound of detail 9, wherein the additional therapeutic agent is a hormone or endocrine therapy agent selected from: a breast / estrogen-driven agent, Tamoxifen, Toremifene, Letrozole, Anastrozole, Exemestane, Fulvestrant, a prostate / androgen-driven agent, Leuprolide, Goserelin, Triptorelin, Degarelix, Abiraterone, Enzalutamide, Apalutamide, and Darolutamide.

[0196] Detail 15: The compound of detail 9, wherein said monoclonal antibody or antibody-drug conjugate is selected from: anti-HER2 antibodies trastuzumab, pertuzumab, margetuximab, trastuzumab emtansine (T-DM1) and trastuzumab deruxtecan (Enhertu); anti-VEGF antibodies bevacizumab and ramucirumab; anti-EGFR antibodies cetuximab and panitumumab; anti-CD20 antibodies rituximab, obinutuzumab and ofatumumab; and antibody-drug conjugates polatuzumab vedotin, brentuximab vedotin, inotuzumab ozogamicin and gemtuzumab ozogamicin.

[0197] Detail 16: The compound of detail 9, wherein the additional therapeutic agent is an epigenetic agent and / or modulator selected from a DNA methyltransferase inhibitor, Azacitidine, Decitabine, an HDAC inhibitor, Vorinostat, Romidepsin, Belinostat, and Panobinostat.

[0198] Detail 17: The compound of detail 9, wherein the additional therapeutic agent is a cell-based immunotherapy selected from: Tisagenlecleucel, Axicabtagene ciloleucel, Lisocabtagene maraleucel, Brexucabtagene autoleucel, Idecabtagene vicleucel, and Ciltacabtagene autoleucel.

[0199] Detail 18: The compound of detail 1 , wherein L is -CH2-CH2-.

[0200] Detail 19: The compound of any preceding detail, wherein the compound of formula (I) is a compound selected from the group consisting of any one of Structures A-D]:Docket No.: 411037-501001 WO[ rucure },Docket No.: 411037-501001 WO[Structure D], and / or structures with IIIPAC names in FIG. 7; wherein R is selected from H, Ci-6alkyl, Ci-6hydroxyalkyl, Ci-4fluoroalkyl, C3-6cycloalkyl, benzyl, phenethyl, phenyl, substituted phenyl, heteroaryl, and / or pyridyl, thienyl, or furyl.

[0201] Detail 20: The compound of any preceding detail, which when administered to a human subject at a therapeutically effective dose exhibits a reduced incidence and / or reduced severity of human toxicities selected from cardiotoxicity, hepatotoxicity, nephrotoxicity, neurotoxicity, myelosuppression, mucositis, gastrointestinal toxicity, dermatologic toxicity, alopecia and peripheral neuropathy, and more particularly exhibits reduced cardiotoxicity, hepatotoxicity, myelosuppression and peripheral neuropathy compared with a reference cytotoxic chemotherapeutic agent.

[0202] Detail 21 : The compound of any preceding detail, wherein the compound is configured to be glycosidase-resistant due to the C-C bond at C-1 (B1) between the glucose and L, whereby the C-1-C(L) C-C bond makes the glycoside glycosidase-resistant.

[0203] Detail 22: A pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of details 1-21 and at least one pharmaceutically acceptable carrier, diluent or excipient.

[0204] Detail 23: The pharmaceutical composition of detail 22, wherein the composition is formulated for administration to a human subject.

[0205] Detail 24: The pharmaceutical composition of detail 23, wherein the composition is formulated for oral administration and is in a solid dosage form selected from a tablet, capsule, granule or powder.

[0206] Detail 25: The pharmaceutical composition of detail 23, wherein the composition is formulated for parenteral administration and is in the form of a sterile solution, suspension or lyophilized powder for reconstitution.Docket No.: 411037-501001 WO

[0207] Detail 26: The pharmaceutical composition of any one of details 22-25, further comprising at least one additional therapeutic agent selected from the additional therapeutic agents recited in detail 9.

[0208] Detail 27: The pharmaceutical composition of detail 26, wherein the at least one additional therapeutic agent is selected from the additional therapeutic agents recited in any one of details 10-17.

[0209] Detail 28: A pharmaceutical combination comprising: (a) a first pharmaceutical composition comprising a compound of any one of details 1-21 and at least one pharmaceutically acceptable carrier, diluent or excipient; and (b) a second pharmaceutical composition comprising at least one additional therapeutic agent selected from the additional therapeutic agents recited in any one of details 10-17; wherein the first and second pharmaceutical compositions are adapted for concurrent, sequential or separate administration.

[0210] Detail 29: A pharmaceutical composition comprising: (a) a therapeutically effective amount of a compound of formula (I) as defined in detail 1 or of any one of details 1-21 ; and (b) at least one pharmaceutically acceptable carrier, diluent or excipient; wherein the composition is suitable for systemic or local administration to a human subject.

[0211] Detail 30: The pharmaceutical composition of detail 29, formulated for intravenous administration and in a dosage form selected from a sterile aqueous solution, a sterile aqueous suspension, and a lyophilized powder for reconstitution.

[0212] Detail 31 : The pharmaceutical composition of detail 29, formulated for oral administration and in a solid dosage form selected from a tablet, capsule, granule and powder.

[0213] Detail 32: The pharmaceutical composition of detail 29, formulated for local administration to a tumor or tumor bed, and in a dosage form selected from an injectable solution, an injectable suspension, a gel, an in s / tu-forming depot, an implant and a biodegradable polymer matrix.

[0214] Detail 33: The pharmaceutical composition of any one of details 29-32, further comprising at least one additional therapeutic agent selected from the additional therapeutic agents recited in any one of details 9-17.

[0215] Detail 34: A kit comprising: (a) a first container containing a compound of any one of details 1-21 or a pharmaceutical composition according to any one of details 29-33; and (b) printed instructions for using the compound or pharmaceutical composition in the treatment of cancer or in reversing a cancer-associated pH gradient in a tumor microenvironment in a subject in need thereof.Docket No.: 411037-501001 WO

[0216] Detail 35: The kit of detail 34, further comprising at least one additional therapeutic agent selected from the additional therapeutic agents recited in any one of details 9-17, optionally contained in a second container.

[0217] Detail 36: Use of a compound of any one of details 1-21 in the manufacture of a medicament for reversing a cancer-associated pH gradient in a tumor microenvironment in a subject in need thereof.

[0218] Detail 37: Use of a compound of any one of details 1-21 in the manufacture of a medicament for treating cancer in a subject in need thereof, wherein the cancer is characterized by a cancer-associated pH gradient.

[0219] Detail 38: Use of a compound of any one of details 1-21 in the manufacture of a medicament for treating cancer in combination therapy, wherein the medicament is to be administered together with at least one additional therapeutic agent selected from the additional therapeutic agents recited in any one of details 9-17.

[0220] Detail 39: A method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formula (I), thereby reversing an abnormal intracellular-to-extracellular pH gradient in cancer cells and inducing apoptosis in the cancer cells.

[0221] Detail 40: The method of detail 39, wherein the cancer is characterized by extracellular acidification and intracellular alkalinization relative to corresponding non- malignant cells.

[0222] Detail 41 : The method of detail 39 or 40, wherein administering the compound of formula (I) increases extracellular pH in a tumor microenvironment, decreases intracellular pH of the cancer cells, or both, such that the intracellular-to-extracellular pH gradient of the cancer cells approaches that of non-malignant cells.

[0223] Detail 42: The method of any one of details 39-41 , wherein inducing apoptosis in the cancer cells is evidenced by activation of caspase-3 / 7, DNA fragmentation, loss of mitochondrial membrane potential, or loss of clonogenic survival.

[0224] Detail 43: The method of any one of details 39-42, wherein the cancer is a solid tumor.

[0225] Detail 44: The method of detail 43, wherein the solid tumor is selected from breast cancer, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, ovarian cancer and glioblastoma.

[0226] Detail 45: The method of any one of details 39-42, wherein the cancer is breast cancer.

[0227] Detail 46: The method of detail 45, wherein the breast cancer comprises estrogen receptor-positive cells of the MCF-7 type or an MCF-7-like tumor.Docket No.: 411037-501001 WO

[0228] Detail 47: The method of any one of details 39-42, wherein the cancer comprises tumor cells grown or capable of growing as multicellular spheroids or three- dimensional aggregates.

[0229] Detail 48: The method of any one of details 39-47, wherein the compound of formula (I) is administered intratumorally, peri-tumorally, intraperitoneally, orally or intravenously.

[0230] Detail 49: The method of any one of details 39-48, further comprising administering to the subject at least one additional therapeutic agent selected from: a cytotoxic chemotherapeutic agent, a targeted small-molecule kinase inhibitor, an immune checkpoint inhibitor, a PARP inhibitor, a hormone or endocrine therapy agent, a monoclonal antibody or antibody-drug conjugate, an epigenetic agent or modulator, and a cell-based immunotherapy.

[0231] Detail 50: A compound of formula (I) for use in treating cancer in a subject in need thereof.

[0232] Detail 51 : The compound for use according to detail 50, wherein the cancer is characterized by an abnormal intracellular-to-extracellular pH gradient comprising extracellular acidification and intracellular alkalinization relative to corresponding non- malignant cells.

[0233] Detail 52: The compound for use according to detail 51, wherein treatment with the compound reverses the abnormal intracellular-to-extracellular pH gradient towards that of non-malignant cells and induces apoptosis in the cancer cells.

[0234] Detail 53: The compound for use according to any one of details 50-52, wherein the cancer is selected from breast cancer, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, ovarian cancer and glioblastoma.

[0235] Detail 54: The compound for use according to any one of details 50-53, wherein the cancer is breast cancer comprising estrogen receptor-positive cells of the MCF- 7 type or an MCF-7-like tumor.

[0236] Detail 55: The compound for use according to any one of details 50-54, wherein the compound is to be administered together with at least one additional therapeutic agent selected from: a cytotoxic chemotherapeutic agent, a targeted small-molecule kinase inhibitor, an immune checkpoint inhibitor, a PARP inhibitor, a hormone or endocrine therapy agent, a monoclonal antibody or antibody-drug conjugate, an epigenetic agent or modulator, and a cell-based immunotherapy.

[0237] Detail 56: Use of a compound of formula (I) in the manufacture of a medicament for treating a cancer characterized by extracellular acidification and intracellular alkalinization in a subject in need thereof.Docket No.: 411037-501001 WO

[0238] Detail 57: The use according to detail 56, wherein treatment with the medicament reverses the abnormal intracellular-to-extracellular pH gradient of the cancer cells and induces apoptosis in the cancer cells.

[0239] Detail 58: Use of a compound of formula (I) in the manufacture of a medicament for reversing an abnormal intracellular-to-extracellular pH gradient in a cancer cell, tumor or tumor microenvironment.

[0240] Detail 59: The use according to detail 58, wherein the medicament is for treating a solid tumor, optionally a solid tumor selected from breast cancer, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, ovarian cancer and glioblastoma.

[0241] Detail 60: A method of enhancing anti-tumor immune surveillance in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I), thereby increasing extracellular pH in a tumor microenvironment and reversing an abnormal intracellular-to-extracellular pH gradient in cancer cells.

[0242] Detail 61 : The method of detail 60, wherein enhancing anti-tumor immune surveillance comprises increasing infiltration, activation and / or cytotoxic activity of CD8+T cells and / or natural killer (NK) cells within the tumor microenvironment.

[0243] Detail 62: The method of detail 60 or 61 , wherein normalization of extracellular pH relieves metabolic suppression of tumor-infiltrating lymphocytes and increases production of one or more effector molecules selected from interferon-y, granzyme B and perforin.

[0244] Detail 63: The method of any one of details 60-62, wherein the tumor microenvironment prior to treatment is characterized by extracellular acidification and intracellular alkalinization relative to corresponding non-malignant tissue.

[0245] Detail 64: The method of any one of details 60-63, wherein administration of the compound of formula (I) reduces activity and / or expression of one or more extracellular proteases that are activated or stabilized by low pH, optionally selected from matrix metalloproteinases and cathepsins.

[0246] Detail 65: The method of any one of details 60-64, wherein restoring immune surveillance results in a reduction in local tumor invasion, metastatic dissemination or both.

[0247] Detail 66: The method of any one of details 60-65, further comprising administering at least one additional immunomodulatory agent selected from: an immune checkpoint inhibitor, a cancer vaccine, an adoptive cell therapy, an oncolytic virus, a cytokine therapy, and a monoclonal antibody or antibody-drug conjugate.

[0248] Detail 67: The method of detail 66, wherein the immune checkpoint inhibitor is selected from an anti-PD-1 antibody, an anti-PD-L1 antibody, an anti-CTLA-4 antibody and combinations thereof.Docket No.: 411037-501001 WO

[0249] Detail 68: The method of any one of details 60-67, wherein the compound of formula (I) is administered prior to or concurrently with the additional immunomodulatory agent to re-sensitize a tumor that is intrinsically resistant or has acquired resistance to the additional immunomodulatory agent.

[0250] Detail 69: A method of restoring tissue-level homeostasis at a cancer site in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I), thereby reversing a cancer-associated pH gradient and normalizing extracellular pH within the affected tissue.

[0251] Detail 70: The method of detail 69, wherein restoring tissue-level homeostasis comprises at least one of: reducing extracellular matrix degradation, reducing protease- mediated invasion, decreasing local inflammatory cytokine levels, and improving perfusion within the tumor-bearing tissue.

[0252] Detail 71 : The method of detail 69 or 70, wherein reversal of the cancer- associated pH gradient renders cancer cells more susceptible to endogenous regulatory mechanisms, including immune-mediated cytotoxicity and apoptosis.

[0253] Detail 72: The method of any one of details 69-71 , wherein the cancer is a solid tumor selected from breast cancer, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, ovarian cancer and glioblastoma.

[0254] Detail 73: The method of any one of details 69-72, wherein the compound of formula (I) is administered locally to or within the tumor, systemically by intravenous infusion, orally, or in a combination thereof.

[0255] Detail 74: A compound of formula (I) for use in enhancing anti-tumor immune surveillance in a subject having a tumor microenvironment characterized by extracellular acidosis, wherein the compound of formula (I), upon administration, increases extracellular pH and reverses an abnormal intracellular-to-extracellular pH gradient in cancer cells.

[0256] Detail 75: The compound for use according to detail 74, wherein enhancement of anti-tumor immune surveillance comprises increasing the number, viability and / or effector function of tumor-infiltrating CD8+T cells and / or NK cells.

[0257] Detail 76: The compound of formula (I) for use in restoring tissue homeostasis at a cancer site by reversing a cancer-induced pH gradient and reducing extracellular protease activity in the tumor microenvironment.

[0258] Detail 77: The compound for use according to detail 76, wherein restoring tissue homeostasis reduces tumor invasion into adjacent tissue, metastatic spread, or both.

[0259] Detail 78: The compound for use according to any one of details 74-77, for use in combination with at least one immunotherapy selected from: an immune checkpoint inhibitor, a therapeutic cancer vaccine, an adoptive T-cell or NK-cell product, and an oncolytic viral therapy.Docket No.: 411037-501001 WO

[0260] Detail 79: Use of a compound of formula (I) in the manufacture of a medicament for enhancing anti-tumor immune surveillance in a subject by normalizing extracellular pH in a tumor microenvironment.

[0261] Detail 80: Use of a compound of formula (I) in the manufacture of a medicament for restoring tissue-level homeostasis at a cancer site in a subject by reversing a cancer-associated pH gradient and decreasing extracellular protease activity.

[0262] Detail 81 : The use of detail 79 or 80, wherein the medicament is formulated for administration together with, or as part of a treatment regimen that further comprises, an immune checkpoint inhibitor, a cancer vaccine, or an adoptive cell therapy.

[0263] Detail 82: A method of treating cancer-associated inflammation in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I), thereby increasing extracellular pH in an inflamed tumor microenvironment and reducing one or more markers of inflammation associated with the cancer.

[0264] Detail 83: The method of detail 82, wherein the cancer-associated inflammation is a chronic or systemic inflammation characterized by persistently elevated circulating levels of one or more inflammatory mediators selected from tumor necrosis factor- a (TNFa), interleukin-6 (IL-6) and C-reactive protein (CRP).

[0265] Detail 84: The method of detail 82 or 83, wherein treatment with the compound of formula (I) decreases local or systemic levels of one or more pro-inflammatory cytokines selected from TNFa, IL-6, I L-1 and CRP, and / or decreases expression of a senescence-associated secretory phenotype (SASP) in cells within or adjacent to the tumor.

[0266] Detail 85: The method of any one of details 82-84, wherein the cancer is a solid tumor selected from breast cancer, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, ovarian cancer and glioblastoma.

[0267] Detail 86: The method of any one of details 82-85, wherein the subject is aged 45 years or older and exhibits age-associated or age-related chronic inflammation in addition to the cancer-associated inflammation.

[0268] Detail 87: The method of any one of details 82-86, wherein the compound of formula (I) is administered as a pharmaceutical composition according to any one of details 29-33.

[0269] Detail 88: A method of treating a chronic inflammatory or autoimmune condition characterized by an acidic extracellular microenvironment at a site of inflammation in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I), thereby increasing extracellular pH at the site of inflammation and reducing the chronic inflammatory response.Docket No.: 411037-501001 WO

[0270] Detail 89: The method of detail 88, wherein the chronic inflammatory or autoimmune condition is selected from rheumatoid arthritis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, psoriasis, psoriatic arthritis, systemic lupus erythematosus, multiple sclerosis and combinations thereof.

[0271] Detail 90: The method of detail 88 or 89, wherein the chronic inflammatory condition is a chronic inflammation of at least about one month in duration, optionally at least about two or three months in duration.

[0272] Detail 91 : The method of any one of details 88-90, wherein administration of the compound of formula (I) increases extracellular pH in the inflamed tissue from a pathologically acidic value to a value approaching physiological pH, optionally to a pH in the range of about 7.0 to about 7.5.

[0273] Detail 92: The method of any one of details 88-91 , wherein the subject exhibits age-related or age-associated systemic low-level inflammation characterized by a two- to threefold increase in circulating cytokines selected from TNFa, IL-6 and CRP relative to a reference population of younger or healthy subjects.

[0274] Detail 93: The method of any one of details 82-92, further comprising administering to the subject at least one additional anti-inflammatory or immunomodulatory agent selected from: a non-steroidal anti-inflammatory drug (NSAID), a corticosteroid, a conventional disease-modifying anti-rheumatic drug (DMARD), a biologic anti-cytokine agent, and a Janus kinase (JAK) inhibitor.

[0275] Detail 94: The method of any one of details 82-93, wherein the compound of formula (I) is administered locally to the inflamed tissue, systemically, or in a combination of local and systemic administration, optionally by oral, intravenous, subcutaneous, intraperitoneal, intra-articular or intralesional administration.

[0276] Detail 95: Use of a compound of formula (I) in the manufacture of a medicament for treating cancer-associated inflammation in a subject in need thereof, wherein treatment with the medicament increases extracellular pH in an inflamed tumor microenvironment and reduces one or more markers of cancer-associated inflammation.

[0277] Detail 96: The use of detail 95, wherein the medicament is for treating a subject having a solid tumor selected from breast cancer, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, ovarian cancer and glioblastoma and a chronic or systemic cancer-associated inflammation.

[0278] Detail 97: Use of a compound of formula (I) in the manufacture of a medicament for treating a chronic inflammatory or autoimmune condition characterized by an acidic extracellular microenvironment at a site of inflammation in a subject in need thereof.Docket No.: 411037-501001 WO

[0279] Detail 98: The use of detail 97, wherein the chronic inflammatory condition is an age-related or age-associated chronic inflammation in a subject aged 45 years or older, optionally a low-level systemic inflammation characterized by elevated circulating TNFa, IL-6 and / or CRP.

[0280] Detail 99: A pharmaceutical combination comprising: (a) a compound of any one of details 1-21; and (b) at least one reverse transcriptase inhibitor (RTI); wherein the compound of any one of details 1-21 and the at least one RTI are formulated or packaged for concurrent, sequential or separate administration to a subject in need thereof.

[0281] Detail 100: The pharmaceutical combination of detail 99, wherein the at least one RTI is a nucleoside or nucleotide reverse transcriptase inhibitor (NRTI).

[0282] Detail 101: The pharmaceutical combination of detail 99 or 100, wherein the at least one RTI is selected from lamivudine, emtricitabine, tenofovir, abacavir, zidovudine, stavudine and combinations thereof.

[0283] Detail 102: The pharmaceutical combination of any one of details 99-101 , wherein the at least one RTI is selected from censavudine, elvucitabine and combinations thereof.

[0284] Detail 103: The pharmaceutical combination of any one of details 99-102, further comprising at least one additional therapeutic agent selected from the additional therapeutic agents recited in any one of details 9-17.

[0285] Detail 104: The pharmaceutical combination of any one of details 99-103, wherein the pharmaceutical combination is for use in treating cancer or a cancer-associated inflammatory disorder in a subject in need thereof.

[0286] Detail 105: The pharmaceutical combination of any one of details 99-103, wherein the pharmaceutical combination is for use in treating age-associated inflammation or a chronic inflammatory condition in a subject in need thereof.

[0287] Detail 106: A method of treating cancer or a cancer-associated inflammatory disorder in a subject in need thereof, the method comprising co-administering to the subject: (a) a therapeutically effective amount of a compound of formula (I); and (b) a therapeutically effective amount of at least one reverse transcriptase inhibitor (RTI); wherein the compound of formula (I) is administered in an amount effective to reverse an abnormal intracellular-to- extracellular pH gradient in cancer cells or in a tumor microenvironment, and the at least one RTI is administered in an amount effective to reduce RTI-sensitive inflammatory signaling.

[0288] Detail 107: The method of detail 106, wherein the at least one RTI is a nucleoside or nucleotide reverse transcriptase inhibitor (NRTI).

[0289] Detail 108: The method of detail 106 or 107, wherein the at least one RTI is selected from lamivudine, censavudine, elvucitabine and combinations thereof.Docket No.: 411037-501001 WO

[0290] Detail 109: The method of any one of details 106-108, further comprising administering to the subject at least one additional therapeutic agent selected from the additional therapeutic agents recited in any one of details 9-17.

[0291] Detail 110: The method of detail 109, wherein the at least one additional therapeutic agent is selected from an immune checkpoint inhibitor, a monoclonal antibody or antibody-drug conjugate, and a cell-based immunotherapy.

[0292] Detail 111: The method of any one of details 106-110, wherein the cancer is a solid tumor, optionally selected from breast cancer, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, ovarian cancer and glioblastoma.

[0293] Detail 112: The method of any one of details 106-111, wherein the compound of formula (I) and the at least one RTI are administered according to a regimen in which the compound of formula (I) is given prior to or concurrently with the at least one RTI so as to normalize extracellular pH and reduce chronic inflammatory signaling within a tumor microenvironment.

[0294] Detail 113: The method of any one of details 106-112, wherein the at least one RTI is administered at a daily dose that is less than or equal to about 80% of a standard adult monotherapy dose for treatment of viral infection, while maintaining a therapeutically effective level for modulation of age- or cancer-associated inflammation.

[0295] Detail 114: A method of treating an age-associated or chronic inflammatory condition characterized by an acidic local microenvironment in a subject in need thereof, the method comprising co-administering to the subject: (a) a therapeutically effective amount of a compound of formula (I); and (b) a therapeutically effective amount of at least one reverse transcriptase inhibitor (RTI); wherein the compound of formula (I) is administered in an amount effective to increase local extracellular pH and / or decrease intracellular pH in affected tissue, and the at least one RTI is administered in an amount effective to reduce reverse-transcriptase-dependent nucleic acid signaling associated with the inflammatory condition.

[0296] Detail 115: The method of detail 114, wherein the age-associated or chronic inflammatory condition is selected from age-associated systemic inflammation, cancer- associated inflammation, osteoarthritis, atherosclerosis, neurodegenerative disease, and chemotherapy-induced chronic inflammation.

[0297] Detail 116: The method of detail 114 or 115, wherein the inflammatory condition is an autoimmune or immune-mediated disorder selected from multiple sclerosis, rheumatoid arthritis, psoriatic arthritis, inflammatory bowel disease, and systemic lupus erythematosus.

[0298] Detail 117: A compound of formula (I) in combination with at least one reverse transcriptase inhibitor (RTI) for use in treating cancer or a cancer-associated inflammatoryDocket No.: 411037-501001 WO disorder in a subject in need thereof, wherein the compound of formula (I), upon administration, reverses an abnormal intracellular-to-extracellular pH gradient in cancer cells and the at least one RTI reduces RTI-sensitive inflammatory signaling.

[0299] Detail 118: A compound of formula (I) in combination with at least one reverse transcriptase inhibitor (RTI) for use in treating an age-associated or chronic inflammatory condition characterized by an acidic microenvironment in a subject in need thereof.

[0300] Detail 119: Use of: (a) a compound of formula (I); and (b) at least one reverse transcriptase inhibitor (RTI); in the manufacture of a medicament for treating cancer or a cancer-associated inflammatory disorder in a subject in need thereof.

[0301] Detail 120: Use of: (a) a compound of formula (I); and (b) at least one reverse transcriptase inhibitor (RTI); in the manufacture of a medicament for treating an age- associated or chronic inflammatory condition characterized by an acidic microenvironment in a subject in need thereof.

[0302] Detail 121: A kit comprising: (a) a first container containing a compound of any one of details 1-21 or a pharmaceutical composition according to any one of details 22- 33; (b) a second container containing at least one reverse transcriptase inhibitor (RTI); and (c) printed instructions for administration of the contents of the first and second containers as combination therapy for treating cancer, a cancer-associated inflammatory disorder, an age- associated inflammatory condition or a chronic inflammatory condition characterized by an acidic microenvironment.

[0303] Detail 122: The kit of detail 121 , wherein the at least one RTI in the second container is selected from lamivudine, censavudine, elvucitabine and combinations thereof.

[0304] Detail 123: A method of selectively killing cancer cells in vitro, the method comprising contacting a population of cancer cells with an effective amount of a compound of formula (I) under conditions sufficient to reverse an abnormal intracellular-to-extracellular pH gradient in the cancer cells and thereby induce apoptosis in the cancer cells.

[0305] Detail 124: The method of detail 123, wherein the cancer cells are contacted with the compound of formula (I) in the presence of one or more non-malignant cells, and the compound of formula (I) selectively reduces viability of the cancer cells relative to the non- malignant cells.

[0306] Detail 125: The method of detail 123 or 124, further comprising detecting a change in intracellular and / or extracellular pH using a pH-sensitive fluorescent indicator before and after contacting the cells with the compound of formula (I).

[0307] Detail 126: The method of any one of details 123-125, wherein the cancer cells are breast cancer cells, optionally estrogen receptor-positive MCF-7 cells or MCF-7-like cells.Docket No.: 411037-501001 WO

[0308] Detail 127: The method of any one of details 123-126, wherein the cancer cells are cultured as multicellular spheroids or three-dimensional aggregates.

[0309] Detail 128: The method of any one of details 123-127, wherein the contacting step is carried out for a period of from about 1 hour to about 96 hours and at a concentration of the compound of formula (I) in the range of about 0.01 pM to about 100 pM.

[0310] Detail 129: A method for assessing or characterizing the sensitivity of cancer cells to a compound of formula (I), the method comprising: (a) providing cancer cells in vitro',(b) contacting the cancer cells with a test concentration of the compound of formula (I); and(c) measuring at least one readout selected from cell viability, apoptosis, intracellular pH and extracellular pH; wherein a decrease in cell viability or an increase in apoptosis accompanied by reversal of an abnormal intracellular-to-extracellular pH gradient indicates sensitivity of the cancer cells to the compound of formula (I).

[0311] Detail 130: The method of detail 129, further comprising contacting a parallel culture of the cancer cells with a reference anti-cancer agent and comparing the readout obtained with the compound of formula (I) to the readout obtained with the reference anticancer agent.

[0312] Detail 131: The method of detail 130, wherein the reference anti-cancer agent is doxorubicin.

[0313] Detail 132: The method of any one of details 129-131 , wherein intracellular or extracellular pH is measured using a pH-sensitive fluorescent indicator, optionally pH rodo or a pH rodo-based probe.

[0314] Detail 133: A method for manufacturing a medicament or cell-based product, the method comprising: (a) providing tumor cells ex vivo, (b) contacting the tumor cells with an effective amount of a compound of formula (I) under conditions that induce apoptosis in the tumor cells by reversing an abnormal intracellular-to-extracellular pH gradient; and (c) formulating the apoptotic tumor cells, fragments thereof or components derived therefrom together with at least one pharmaceutically acceptable carrier, diluent or excipient to obtain the medicament or cell-based product.

[0315] Detail 134: The method of detail 133, wherein the tumor cells are autologous tumor cells obtained from a subject to be treated.

[0316] Detail 135: The method of detail 133 or 134, wherein the medicament or cellbased product is a tumor cell vaccine or an immunogenic composition comprising apoptotic tumor cells or fragments thereof.

[0317] Detail 136: The method of any one of details 133-135, further comprising combining the apoptotic tumor cells, fragments thereof or components derived therefrom with at least one immunostimulatory adjuvant.Docket No.: 411037-501001 WO

[0318] Detail 137: A kit for assessing sensitivity of cancer cells to a pH-gradient- reversing agent, the kit comprising: (a) a compound of formula (I); and (b) a pH-sensitive fluorescent indicator suitable for measuring intracellular and / or extracellular pH in living cells.

[0319] Detail 138: The kit of detail 137, wherein the pH-sensitive fluorescent indicator comprises a pH rodo-based fluorophore.

[0320] Detail 139: The kit of detail 137 or 138, further comprising at least one container or plate suitable for culturing cancer cells in monolayer or as three-dimensional spheroids.

[0321] Detail 140: The kit of any one of details 137-139, further comprising printed instructions describing a protocol for contacting cancer cells with the compound of formula (I), detecting changes in pH using the pH-sensitive fluorescent indicator, and determining sensitivity of the cancer cells to the compound of formula (I) based on the detected changes.

[0322] Detail 141 : A kit for in vitro screening of candidate pH-gradient-modulating agents, the kit comprising: (a) a compound of formula (I) as a positive control; (b) a pH- sensitive fluorescent indicator suitable for measuring intracellular and / or extracellular pH; and (c) instructions for comparing pH changes and / or cell viability obtained with one or more test agents to those obtained with the compound of formula (I) to identify candidate pH- gradient-modulating agents.

[0323] Detail 142: A method of modulating an intracellular-to-extracellular pH gradient in a cell in vitro, the method comprising contacting the cell with an effective amount of a compound of formula (I) under conditions sufficient to increase extracellular pH and / or decrease intracellular pH in the cell.

[0324] Detail 143: The method of detail 142, wherein the cell is a cancer cell, an immune cell or a stromal cell obtained from a tumor or from inflamed tissue.

[0325] Detail 144: A method for selecting a treatment regimen for a subject having cancer, the method comprising: (a) obtaining tumor cells from the subject; (b) contacting the tumor cells ex vivo with a test concentration of a compound of formula (I); (c) determining, following step (b), at least one change selected from a decrease in cell viability, an increase in apoptosis, an increase in extracellular pH and a decrease in intracellular pH in the tumor cells; and (d) on the basis of the at least one change determined in step (c), selecting a treatment regimen for the subject that comprises administration of a therapeutically effective amount of a compound of formula (I).

[0326] Detail 145: The method of detail 144, wherein the treatment regimen further comprises administration of at least one additional therapeutic agent selected from the additional therapeutic agents recited in any one of details 9-17.Docket No.: 411037-501001 WO

[0327] Detail 146: The method of any one of details 39-49, wherein the compound of formula (I) is administered at a dose in the range of about 0.01 to about 100 mg / kg body weight per day.

[0328] Detail 147: The method of detail 146, wherein the compound of formula (I) is administered at a dose in the range of about 0.1 to about 20 mg / kg body weight per day.

[0329] Detail 148: The method of any one of details 82-94, wherein the compound of formula (I) is administered at a dose in the range of about 0.01 to about 100 mg / kg body weight per day.

[0330] Detail 149: The method of any one of details 39-49 or 82-94, wherein the compound of formula (I) is administered once daily, twice daily, or intermittently three times per week.

[0331] Detail 150: The method of any one of details 39-49, further comprising administering radiotherapy to the subject, wherein the radiotherapy is directed to a tumor and the compound of formula (I) is administered before, during or after the radiotherapy.

[0332] Detail 151: Use of a compound of formula (I) in the manufacture of a medicament for treating cancer in a subject in need thereof, wherein the medicament is for administration in combination with radiotherapy and is effective to reverse an abnormal intracellular-to-extracellular pH gradient in cancer cells and to improve the response of the cancer to the radiotherapy.

[0333] Detail 152: A kit comprising: (a) a first container containing a compound of any one of details 1-21 or a pharmaceutical composition according to any one of details 22- 33; and (b) printed instructions for administering the compound or pharmaceutical composition to a subject in combination with radiotherapy for the treatment of cancer.

[0334] Detail 153: A method of reducing the risk of recurrence or metastatic spread of a cancer in a subject who has previously received a primary cancer therapy, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I) after completion of the primary cancer therapy, thereby reversing a cancer- associated pH gradient in residual cancer cells or in a tumor bed and reducing the risk of recurrence or metastasis.

[0335] Detail 154: The method of detail 153, wherein the primary cancer therapy comprises surgery, radiotherapy, chemotherapy, immunotherapy or any combination thereof.

[0336] Detail 155: The method of detail 153 or 154, wherein the cancer is a solid tumor selected from breast cancer, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, ovarian cancer and glioblastoma.

[0337] In some embodiments, the technology provides two different “levels” of intervention against disease. At the microcosm level, which is associated with chronic inflammation and RTI (reverse transcriptase inhibitor) approaches, drugs block or inhibitDocket No.: 411037-501001 WO specific molecular pathways — like “throwing a wrench” into one gear of the machine — but cancer and autoimmune diseases are so plastic that they often route around that block. At the macrocosm level, the technology is focused on broad, integrated properties of cancer, especially its altered metabolic profile and abnormal pH environment. The technology enables that, instead of chasing individual signaling pathways, an approach should manipulate the metabolic context itself, for example by manipulating the metabolic context itself, for example by normalizing extracellular pH in tissues that chronic inflammation has made acidic, including by redistributing protons into intracellular or organellar compartments while driving the bulk extracellular milieu toward physiological values. In this view, shifting this metabolic and pH landscape makes cancer cells far more vulnerable to normal control mechanisms such as immune surveillance and apoptosis, and cancer has a harder time adapting to these “environmental” changes than to single-pathway inhibitors. In some embodiments, macrocosm-level pH modulation is implemented alone or in combination with microcosm-level agents, such as reverse transcriptase inhibitors, to provide complementary control of both metabolic context and pathway-specific signaling.

[0338] In some embodiments, cancer cells are understood to establish an abnormal intracellular-to-extracellular pH gradient in which the intracellular space becomes slightly alkaline and the extracellular milieu becomes acidic or highly acidic, in contrast to normal cells where the intracellular pH is slightly acidic and the extracellular pH is slightly alkaline. This cancer-induced gradient interferes with T-cell metabolic activity, facilitates immune evasion, and promotes tumor invasion and metastasis through enhanced activity of extracellular proteases that disrupt cell-matrix adhesion and increase cellular motility. According to some aspects, this state represents a pathological “cancer homeostasis” that supports malignant growth and dissemination.

[0339] In some embodiments, the technology disrupts this cancer homeostasis by reversing the abnormal pH gradient back toward normal values. According to some aspects, the disclosed molecules act as base-conjugated, glycosidase-resistant C-linked carbohydrates that move protons in the opposite direction to that demanded by the cancer equilibrium, thereby undermining the cancer cell’s ability to maintain its preferred pH balance. This proton transfer leads cancer cells toward apoptosis and restores the natural homeostasis of the tissue at the cancer site. In a non-limiting example, the constructs include hybrid molecules in which a glucose C-glycoside is covalently linked through a C- linkage to a lysine or arginine amino acid residue, with an optional substituent R on the linker methylene, where R can be hydrogen, methyl, or phenyl. The C-linkage increases molecular stability and prevents glycolysis of the sugar moiety, thereby supporting sustained pH- modulating activity. According to some aspects, this platform is characterized as anDocket No.: 411037-501001 WO innovative cancer-targeted pH-gradient-reversing technology that induces apoptosis of cancer cells and re-establishes physiological tissue homeostasis at the tumor site.

[0340] Compatibility qualifier: All combinations are intended “except where technically incompatible.” A combination is technically incompatible only if two features cannot reasonably be implemented together by a skilled person using routine techniques and without undue burden, having regard to ordinary workshop modifications and substitutions (for example, material choice, scale, or type of fastener or textile). Absent such incompatibility, any feature described herein may be combined with any other feature, even if originally disclosed in different sections, figures, or examples.

[0341] Support for amendments: The foregoing sentences are intended to provide explicit written basis for: (i) selecting one or more members from each of two or more lists to form a specific combination; (ii) combining features originally disclosed in different examples, figures, or sections of the description and drawings; (iii) generating narrowed sub-ranges and discrete values from disclosed ranges; (iv) migrating a feature from a dependent to an independent claim (and vice versa); and (v) recasting the same technical teaching across apparatus, method, kit, and use-related claim formats. Unless expressly stated to the contrary, every embodiment disclosed herein is intended to be combinable, directly and without further invention, with every other embodiment to the fullest extent permitted by the skilled person’s ordinary practice.

[0342] The features, integers, characteristics, compounds, chemical structures, linkers, amino acids, groups, pH ranges, dosage ranges, formulation types, routes of administration, indications, and method steps disclosed in the present specification, drawings and examples may be combined in any manner that is technically compatible, unless the context clearly dictates otherwise. In some embodiments, any feature or embodiment described in connection with one aspect of the disclosure is equally applicable to, and combinable with, any other aspect or embodiment described herein. The various embodiments and alternatives disclosed herein are therefore to be understood as individually disclosed and as combinable in any operative combination.

[0343] According to some aspects, individual features described in the context of a particular compound of formula (I), a particular linker L, a particular amino acid residue AA, a particular dosage form, a particular indication (for example, cancer, cancer-associated inflammation, chronic inflammatory disorders, autoimmune diseases, or age-associated systemic inflammation), or a particular route or schedule of administration, may be combined with any other compound, linker, amino acid residue, dosage form, indication, route or schedule of administration described herein, provided that the resulting combination is technically feasible. Unless explicitly stated otherwise, such combinations are intended to be encompassed within the scope of the present disclosure.Docket No.: 411037-501001 WO

[0344] In some embodiments, features described in the Examples are not limited to the specific combinations explicitly set out, but may be recombined with features of any other Example, figure, paragraph or embodiment of the invention. For example, any dosing regimen, route of administration, formulation, animal model, in vitro assay, pH-measurement technique, or biomarker readout described in one Example may be used with any compound of formula (I), indication or therapeutic combination described in another Example or elsewhere in the specification, to the extent that such combinations are technically compatible. The Examples are therefore to be understood as illustrative and non-limiting, and the invention encompasses all combinations and sub-combinations of features disclosed herein.

[0345] The technology described herein is further illustrated by the following examples which in no way should be construed as being further limiting. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below.EXAMPLES

[0346] The invention now being generally described, it will be more readily understood by reference to the following examples which are included merely for purposes of illustration of certain aspects and embodiments of the present invention and are not intended to limit the invention.EXAMPLE 1 PREPARATION OF COMPOUNDS

[0347] The following prophetic example illustrates the preparation of compounds by a multi-step process. In a non-limiting prophetic example, representative compounds of formula (I), G-L-NH-CO-AA, are prepared using a convergent strategy that assembles a glycosidase-resistant glucose C-glycosyl residue, a linker segment L, and a basic amino acid residue AA. In some embodiments, a protected D-glucose or L-glucose precursor is converted to a C-glycosyl intermediate by forming a carbon-carbon bond between the anomeric carbon (C-1) and a suitably functionalized methylene or alkylene fragment destined to become part of linker L. According to some aspects, this step can be accomplished by reacting a protected sugar lactone, glycal, or aldehyde with an organometallic or other carbon-based nucleophile under conditions that provide the desired C-glycosidic architecture while preserving stereochemical integrity at the remaining sugar centers.

[0348] In some embodiments, the C-glycosyl intermediate is subsequently elaborated to introduce a terminal amine, carboxylic acid, or activated ester that defines the remaining portion of linker L. For example, in a non-limiting example, hydrogenolysis and / or acidolysis of protecting groups followed by functional group interconversion furnishes a C-Docket No.: 411037-501001 WO glycosyl linker building block bearing an N-protected amino functionality suitable for peptide coupling. According to some aspects, this intermediate is then coupled to an N-protected basic amino acid, such as Boc-L-lysine or Boc-L-arginine, using standard peptide coupling reagents (e.g., carbodiimide or uronium reagents) to afford an amide of the general G-L-NH- CO-AA type.

[0349] In this example, global deprotection of temporary protecting groups on the sugar hydroxyls and amino acid side chain, followed by purification by chromatography and / or crystallization, yields the target compound of formula (I) as a free base or pharmaceutically acceptable salt. In some embodiments, at least one hydroxyl group on the glucose residue is optionally acylated to provide a prodrug ester. According to some aspects, this general route can be adapted to prepare a panel of analogs that differ in sugar configuration (D or L), linker length and composition, or the identity of the basic amino acid residue AA.

[0350] The technology can be made of any material suitable to practice the methods disclosed herein. Small molecules are envisioned as discussed above along with salts, hydrates, solvates, and / or prodrugs. The present disclosure encompasses the preparation and use of solvates in small molecules and also methods to remove water and / or solvent from small molecules. Solvates typically do not significantly alter the properties or toxicity of the small molecules, and as such may function as equivalents. The term "solvate" as used herein is a combination, physical association and / or solvation of a compound of the present disclosure with a solvent molecule such as, e.g., a disolvate, monosolvate or hemisolvate, where the ratio of solvent molecule to compound of the present disclosure can be about 2:1 , about 1 : 1 or about 1 :2, respectively (per unit small molecule in this non-limiting example). This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances, the solvate can be isolated, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. Thus, "solvate" encompasses both solution-phase and isolatable solvates. One type of solvate is a hydrate. A "hydrate" relates to a particular subgroup of solvates where the solvent molecule is water. Solvates typically can function as equivalents. Preparation of solvates is known in the art. For example, a small molecule is dissolved in water and methanol (or other solvent), dried, and the resulting O-H stretch (from the formation of the solvate / hydrate) is confirmed by an ATR-IR measurement. The O-H stretch is not found in the non-solvate / hydrate form, quickly confirming the formation of the solvate / hydrate in the solid form by the ATR-IR measurement. A solvate can be used (e.g., in formulations) to enhance the solubility of a small molecule.Docket No.: 411037-501001 WOEXAMPLE 2 CHEMICAL TARGET 4 RESULTS WITH TARGET 3 DATA

[0351] In this example, Chemical Target 4 is the chemical compound with structure depicted in FIG. 9 and in the MS spectra of FIGs. 11A-11B (with chromatogram in FIG. 10).

[0352] Regarding Chemical Target 3 (see FIG. 8 for chemical structure), in some embodiments, FIG. 8 schematically illustrates the glycosidase-resistant D-glucose C- glycoside-amino-acid hybrid corresponding to Chemical Target 3. The figure depicts a D- glucose ring in the pyranose form with a carbon-carbon bond between the anomeric carbon and a two-carbon linker terminating in a secondary amine that forms an amide with the side chain of a basic amino acid residue, for example a lysine- or arginine-type residue. According to some aspects, this scaffold exemplifies the class of C-glycoside amino-acid conjugates evaluated in this, and FIG. 7 also lists IIIPAC names for non-limiting example compounds sharing this architecture. In this example NMR spectrum of FIG. 8, the spectrum is used to highlight the modular arrangement of the sugar, linker, and basic amino-acid moieties and to illustrate positions that can be modified without disrupting the C-glycosidic linkage. Although Chemical Target 4 is the compound tested in Chemical Target 4 experiments, Chemical Target 3 provides an archetypal structure that aids in interpreting the Chemical Target 4 biological data.

[0353] A cytotoxicity assay of Chemical Target 4 in an MCF-7 cell line was conducted. FIGs. 1A-1B show results of the cell viability assessment by means of a 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl-2H-" bromide (MTT) assay for Chemical Target 4 following 48h incubation in the MCF-7 cell line. Control cells were administered DMSO in concentrations matching those delivered with the compound. The graphs in FIGs. 1A-1B depict the absorbance at 570 nm determined for MCF-7 cell line following treatment with the indicated compound at lower (FIG. 1A) and higher (FIG. 1B) gradually increasing concentrations. FIG. 1A shows (0, 6.25, 12.5, 25, 50, 100 and 200 pg / ml) concentrations and FIG. 1B shows (0, 50, 100, 200, 500 and 1000 pg / ml) concentrations. The background absorbance of the plates at 630 nm was also measured and subtracted from 570 nm measurement.

[0354] A pH Rodo Assay of Chemical Target 4 in MCF-7 Cell Line was performed and measured. FIG. 2A shows images from the measurements of intracellular pH using pH Rodo indicators for the determination of fluorescence intensity for intracellular pH values at 24h post-treatment for Chemical Target 4 in the MCF-7 cell line. FIG. 2B shows the plot of target 4 concentration (pg / mL) versus mean fluorescence intensity. Control cells were administered DMSO in concentrations matching those delivered with the compound. In these examples, in some embodiments, the pH rodo = compartment / endo-Docket No.: 411037-501001 WO lysosomal / pericellular readout; and bulk extracellular pH normalization is inferred and supported by the mechanistic model and in vivo prophetic examples, not by pH rodo alone.

[0355] FIG. 3 shows images and results from the Cell Event™ Caspase-3 / 7 Assay. The control MCF-7 Cells are the first 3 columns from left, and the treated MCF-7 cells are the 3 columns at right. First row is Day -6 (1stday after plating 5000 MCF-7 cells / well); the second row is day 0 (spheroids). The third row is day 2 (after 48h treatment), and the fourth row is day 2 (CellEvent™ Caspase-3 / 7 Assay at 200pm). FIG. 4 shows images from the pH Rodo Assay.

[0356] FIG. 5A shows a side-by-side comparison of images from the CellEvent™ Caspase-3 / 7 Assay and from the pH Rodo Assay. For the CellEvent™ Caspase-3 / 7 Assay, FIG. 5B shows a plot of (X-axis) the control and treated MCF-7 cells versus (Y-axis) mean fluorescence intensity (200 pm, see bottom of FIG. 5A). For the pH Rodo Assay, FIG. 5C shows a plot of (X-axis) the control and treated MCF-7 cells versus (Y-axis) mean fluorescence intensity (200 pm, see bottom of FIG. 5A).

[0357] For MCF-7 breast adenocarcinoma cells, FIG. 6A is a conceptual overview of the 3-D MCF-7 spheroid model and the treatment outcome. Left side (FIG. 6A): shows how MCF-7 breast adenocarcinoma cells start as loose aggregates (day 1), then tight aggregates (day 2), and by day 7 form a smooth, compact spheroid with typical zones: a proliferative outer shell, a quiescent middle layer and a necrotic core. The arrow labelled “IC3048-hour treatment - potential anti-cancer peptide” indicates that the mature spheroids were treated for 48 h at the IC30concentration of Chemical Target 4. Right side (FIG. 6A): the final “irregular decreased spheroid size” graphic, with symbols for viable, dying and dead cancer cells, illustrates that treatment makes the spheroid smaller and more disorganized, with many cells undergoing death. In some embodiments, FIG. 6A is summarizing experimental design + biological interpretation whereby a mature 3-D tumor spheroid is exposed to the compound, which leads to shrinkage and a higher fraction of dying / dead cells.

[0358] FIG. 6B (the micrographs: brightfield, green and red images) shows the actual microscopy data that back up that schematic in FIG. 6A. Top row (FIG. 6B): brightfield images of untreated vs treated spheroids. The treated spheroid is smaller and less smooth, consistent with partial collapse or loss of cells. Bottom left pair (green or grey, FIG. 6B): CellEvent™ caspase-3 / 7 fluorescence. Green (or grey) signal marks cells with activated executioner caspases (apoptosis). Treated spheroids show stronger, more widespread green (or grey) signal than untreated, indicating more apoptotic cells. Bottom right pair (red or grey, FIG. 6B): pH rodo fluorescence, which increases in more acidic environments. Treated spheroids show brighter red (or grey) staining than untreated, indicating increased local acidification (consistent with the pH-modulation / proton-handling mechanism central to the technology).Docket No.: 411037-501001 WO

[0359] In some embodiments, together, FIG. 6B is illustrating that Chemical Target 4 makes the 3-D spheroids shrink, increases caspase-3 / 7-positive apoptotic cells, and increases pH rodo signal, supporting the idea that the compound both perturbs pH and drives cancer-cell death in the spheroid model, while FIG. 6A provides the conceptual schematic.

[0360] In some embodiments, FIG. 9 presents a representative1H nuclear magnetic resonance (NMR) spectrum acquired for a glucose C-glycosyl basic amino acid conjugate that is depicted at the left of the figure together with a schematic formate counterion, indicating isolation of the compound as a formate salt. The disassociated chemical compound is ideal for testing as Chemical Target 4. According to some aspects, the spectrum is recorded under standard one-dimensional proton NMR conditions on a 400 MHz instrument, and the trace on the right spans the chemical-shift region from about 0 to 10 ppm, with acquisition parameters summarized above the spectrum. In this example, the distribution and multiplicity of resonances arising from the glucopyranose ring protons, the methylene units of the carbon-carbon linker, and the a- and side-chain protons of the basic amino acid residue provide confirmation of the expected framework of the glucose C- glycoside-amino-acid hybrid in its formate salt form.

[0361] In some embodiments, FIG. 10 presents an LCMS report for Chemical Target 4, the glucose C-glycosyl basic amino acid conjugate whose structure is shown in FIG. 9. According to some aspects, the figure displays stacked chromatograms acquired on an LCMS system using an evaporative light scattering detector (ELSD) and diode array detectors (DAD) at 220 nm and 254 nm, with an additional mass spectrometric trace recorded in parallel. The upper panel corresponds to the ELSD signal, and the lower panels show the UV traces, which collectively indicate a single, well-defined chromatographic peak at a characteristic retention time for this sample. In this example, the integration results table at the bottom of FIG. 10 summarizes the analysis of the ELSD chromatogram, listing retention time, peak width, height, and area, and reporting essentially 100% of the integrated area for a single peak. These data demonstrate chromatographic homogeneity and support the identity and purity assignment for Chemical Target 4.

[0362] In some embodiments, FIG. 11A presents an electrospray ionization mass spectrum acquired at a retention time of 0.311 minutes for the glucose C-glycoside basic amino acid conjugate whose chemical structure is shown in FIG. 9. According to some aspects, the full-scan spectrum in FIG. 11A displays a predominant ion consistent with the protonated molecular species, together with minor fragment ions attributed to cleavage within the linker region and side chain. In this example, FIG. 11B provides a magnified view of the principal mass-to-charge window encompassing the molecular ion, thereby resolving the isotopic envelope and confirming a clean signal for the target species. FIG. 11B furtherDocket No.: 411037-501001 WO illustrates that no peak consistent with a formate adduct at approximately 45 mass units above the protonated molecular ion is present, supporting assignment of the observed signal to the non-formate form of the glucose C-glycoside-amino-acid hybrid under the LC-MS conditions employed.

[0363] In this Example 2, representative compounds of formula (I), G-L-NH-CO-AA, are evaluated for their ability to modulate cancer-associated pH gradients and reduce tumor cell viability in vitro using human breast adenocarcinoma MCF-7 cells and MCF-7-like cells. In some embodiments, monolayer cultures are exposed to a concentration range of a prototype compound (e.g., Chemical Target 4) for 24-48 hours and cell viability is assessed using a metabolic assay such as MTT or an equivalent tetrazolium-based readout. According to some aspects, treatment with the compound produces a concentrationdependent decrease in absorbance relative to vehicle-treated controls, consistent with reduced mitochondrial activity and loss of viable cells.

[0364] In a non-limiting example, companion assays are conducted to assess intracellular and pericellular pH using a pH-sensitive fluorogenic indicator of the pH rodo type. In some embodiments, increasing concentrations of the compound are associated with progressive elevations in mean pH rodo fluorescence intensity, indicating a redistribution or enhancement of acidic compartments relative to baseline conditions. According to some aspects, the same treatment conditions are further interrogated using a caspase-3 / 7 activation assay, such as CellEvent™, which reveals increased green fluorescence in treated cultures, consistent with induction of apoptosis.

[0365] In some embodiments, three-dimensional MCF-7 spheroid models are included to better approximate solid-tumor architecture. In a non-limiting example, spheroids treated with compounds of formula (I) exhibit disrupted morphology, heightened pH rodo signal, and robust caspase-3 / 7 activation compared with untreated controls, supporting the conclusion that pH-gradient modulation and apoptosis occur under diffusion-limited conditions. According to some aspects, these in vitro findings are consistent with the proposed cancer homeostasis disruption technology, in which glucose C-glycosyl basic amino acid conjugates reverse or normalize abnormal intracellular-to-extracellular pH gradients and thereby compromise tumor cell survival.EXAMPLE S ANIMAL STUDIES

[0366] This prophetic example uses rodent tumor models and tests the following example embodiments. In some embodiments, the anti-cancer and anti-inflammatory activity of compounds of formula (I), G-L-NH-CO-AA, is evaluated in vivo using established rodent tumor models. According to some aspects, a representative compound of formula (I) is selected in which G is a D-glucose C-glycosyl residue, L is an ethylene-oxy or propylene-Docket No.: 411037-501001 WO oxy linker, and AA is a basic amino acid such as L-lysine or L-arginine, corresponding to the species used in the in vitro MCF-7 assays described above. In some embodiments, two or more analogs differing in linker length and AA side-chain basicity are included to provide structure-activity information in vivo.

[0367] In a non-limiting example, female athymic nude mice bearing subcutaneous MCF-7 human breast adenocarcinoma xenografts are used as a first model. MCF-7 cells are implanted into the flank, and animals are randomized into groups (for example 8-10 mice per group) when tumors reach approximately 100-200 mm3. In some embodiments, test groups receive the compound of formula (I) formulated in a pharmaceutically acceptable vehicle and administered by intravenous, intraperitoneal, or oral routes at dose levels spanning, for example, about 0.3 mg / kg, about 3 mg / kg, and about 30 mg / kg once daily or every other day for 21-28 days. Dose selection is guided by in vitro IC50or IC30values, solubility, and a preliminary single-dose tolerability study that identifies a maximum tolerated dose based on body weight change, clinical observations, and serum chemistry.

[0368] According to some aspects, tumor volumes are measured at regular intervals using calipers, and percent tumor growth inhibition relative to vehicle control is calculated at the end of the dosing period. Additional secondary endpoints include animal survival, bodyweight trajectories, and clinical signs of toxicity. In some embodiments, blood is collected at defined time points after dosing for pharmacokinetic analysis by LC-MS / MS, allowing correlation of systemic exposure (for example, area under the curve and Cmax) with antitumor activity and with modulation of extracellular pH at the tumor site.

[0369] In this example, modulation of the abnormal intracellular-to-extracellular pH gradient is assessed using complementary approaches. Tumor interstitial pH can be monitored by pH-sensitive microelectrodes, magnetic resonance-based pH imaging, or implantation of pH-responsive fluorescent probes analogous to those employed in FIGs. 2- 5. Tumors harvested at selected time points are processed for histology and immunohistochemistry, including markers of apoptosis (for example, cleaved caspase-3), proliferation (for example, Ki-67), and pH-regulated transporters or enzymes. In some embodiments, treated tumors exhibit a statistically significant increase in extracellular pH toward physiological values, a relative decrease in intracellular pH, reduced proliferation indices, and increased apoptosis relative to vehicle controls.

[0370] In another non-limiting example, syngeneic immunocompetent mouse models, such as 4T1 breast carcinoma or CT26 colon carcinoma implanted in BALB / c mice, are employed to evaluate effects on immune surveillance and cancer-associated inflammation. According to some aspects, compounds of formula (I) are administered at dose levels and schedules similar to those described above, either as monotherapy or in combination with standard-of-care agents (for example, doxorubicin or immune checkpointDocket No.: 411037-501001 WO inhibitors). Endpoints can include tumor growth, lung metastasis burden, tumor-infiltrating lymphocyte profiles, and cytokine signatures in plasma and tumor tissue, thereby connecting pH-gradient modulation with immune and inflammatory readouts.

[0371] In some embodiments, systemic tolerability is further characterized in non- tumor-bearing rodents given repeated doses of the compound of formula (I) at and above the efficacious exposure identified in the xenograft studies. Clinical chemistry (for example, liver enzymes, blood urea nitrogen, creatinine, electrolytes), hematology, and gross and microscopic examination of major organs are used to identify potential target-organ toxicities and to define dose levels that are well tolerated during subacute administration. These data can be used to refine the “therapeutically effective amount” and to support the dosing ranges recited in the claims.

[0372] According to some aspects, the animal studies described in this prophetic example are intended to evaluate and illustrate the methods of Example embodiments 1-25 described below. In some embodiments, successful modulation of tumor pH gradients, reduction in tumor growth or metastatic spread, improvement in survival, and acceptable safety margins in these models support the use of compounds of formula (I) in treating cancers, cancer-associated inflammation, chronic inflammatory disorders, autoimmune diseases, and age-associated systemic inflammation as set forth in Example embodiments 1-25.

[0373] Example embodiment 1: A method of treating a disease in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a compound of formula (I): G-L-NH-CO-AA, thereby modulating an abnormal intracellular-to-extracellular pH gradient in diseased cells in the subject and treating the disease, wherein the disease is selected from the group consisting of cancer, cancer- associated inflammation, chronic inflammatory disorders, autoimmune diseases, age- associated systemic inflammation, and combinations thereof.

[0374] Example embodiment 2: The method of example embodiment 1 , wherein G is D-glucose or L-glucose, and wherein the D-glucose or L-glucose is linked to the linker L via a glycosidic bond.

[0375] Example embodiment 3: The method of example embodiment 1 , wherein L is a linker selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, and substituted heterocycloalkyl.

[0376] Example embodiment 4: The method of example embodiment 1 , wherein AA is an amino acid selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, aspartic acid, glutamic acid, asparagine, glutamine, lysine, arginine, histidine, phenylalanine, tyrosine, tryptophan, and proline.Docket No.: 411037-501001 WO

[0377] Example embodiment 5: The method of example embodiment 1 , wherein the compound of formula (I) is administered in a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0378] Example embodiment 6: The method of example embodiment 5, wherein the pharmaceutical composition is formulated for oral, parenteral, subcutaneous, intravenous, intramuscular, or topical administration, and wherein the pharmaceutical composition is in a form selected from the group consisting of a tablet, capsule, pill, powder, liquid, suspension, emulsion, gel, ointment, cream, patch, and injectable solution.

[0379] Example embodiment 7: The method of example embodiment 1 , wherein the subject is a human, and wherein the human is an adult, child, or infant.

[0380] Example embodiment 8: The method of example embodiment 1 , wherein the cancer is selected from the group consisting of breast cancer, lung cancer, colon cancer, prostate cancer, liver cancer, pancreatic cancer, brain cancer, ovarian cancer, leukemia, lymphoma, melanoma, renal cancer, bladder cancer, head and neck cancer, and sarcoma.

[0381] Example embodiment 9: The method of example embodiment 1 , wherein the chronic inflammatory disorder is selected from the group consisting of rheumatoid arthritis, inflammatory bowel disease, asthma, chronic obstructive pulmonary disease, psoriasis, atopic dermatitis, and vasculitis.

[0382] Example embodiment 10: The method of example embodiment 1, wherein the autoimmune disease is selected from the group consisting of multiple sclerosis, systemic lupus erythematosus, type 1 diabetes, Graves' disease, Hashimoto's thyroiditis, Sjogren's syndrome, and scleroderma.

[0383] Example embodiment 11 : A method of modulating an abnormal intracellular- to-extracellular pH gradient in diseased cells in a subject, the method comprising: administering to the subject a therapeutically effective amount of a compound of formula (I): G-L-NH-CO-AA, thereby modulating the abnormal intracellular-to-extracellular pH gradient in the diseased cells.

[0384] Example embodiment 12: The method of example embodiment 11 , wherein the diseased cells are cancer cells, and wherein the cancer cells are selected from the group consisting of breast cancer cells, lung cancer cells, colon cancer cells, prostate cancer cells, liver cancer cells, pancreatic cancer cells, brain cancer cells, ovarian cancer cells, leukemia cells, lymphoma cells, melanoma cells, renal cancer cells, bladder cancer cells, head and neck cancer cells, and sarcoma cells.

[0385] Example embodiment 13: The method of example embodiment 11, wherein the diseased cells are cells associated with a chronic inflammatory disorder, and wherein the chronic inflammatory disorder is selected from the group consisting of rheumatoid arthritis,Docket No.: 411037-501001 WO inflammatory bowel disease, asthma, chronic obstructive pulmonary disease, psoriasis, atopic dermatitis, and vasculitis.

[0386] Example embodiment 14: The method of example embodiment 11 , wherein the diseased cells are cells associated with an autoimmune disease, and wherein the autoimmune disease is selected from the group consisting of multiple sclerosis, systemic lupus erythematosus, type 1 diabetes, Graves' disease, Hashimoto's thyroiditis, Sjogren's syndrome, and scleroderma.

[0387] Example embodiment 15: The method of example embodiment 11, wherein the diseased cells are cells associated with age-associated systemic inflammation, and wherein the age-associated systemic inflammation is associated with aging, frailty, or age- related diseases.

[0388] Example embodiment 16: The method of example embodiment 11 , wherein G is D-glucose or L-glucose, and wherein the D-glucose or L-glucose is linked to the linker L via a glycosidic bond.

[0389] Example embodiment 17: The method of example embodiment 11, wherein L is a linker selected from the group consisting of alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, and substituted heterocycloalkyl.

[0390] Example embodiment 18: The method of example embodiment 11, wherein AA is an amino acid selected from the group consisting of glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, aspartic acid, glutamic acid, asparagine, glutamine, lysine, arginine, histidine, phenylalanine, tyrosine, tryptophan, and proline.

[0391] Example embodiment 19: The method of example embodiment 11, wherein the compound of formula (I) is administered in a pharmaceutical composition comprising one or more pharmaceutically acceptable carriers, excipients, or diluents.

[0392] Example embodiment 20: The method of example embodiment 19, wherein the pharmaceutical composition is formulated for oral, parenteral, subcutaneous, intravenous, intramuscular, or topical administration, and wherein the pharmaceutical composition is in a form selected from the group consisting of a tablet, capsule, pill, powder, liquid, suspension, emulsion, gel, ointment, cream, patch, and injectable solution.

[0393] Example embodiment 21 : The method of example embodiment 1 or 11 , wherein the compound of formula (I) modulates the abnormal intracellular-to-extracellular pH gradient by increasing the extracellular pH, decreasing the intracellular pH, or both.

[0394] Example embodiment 22: The method of example embodiment 1 or 11, wherein the compound of formula (I) is administered in combination with one or more additional therapeutic agents selected from the group consisting of chemotherapeutic agents, anti-inflammatory agents, immunosuppressive agents, and anti-autoimmune agents.Docket No.: 411037-501001 WO

[0395] Example embodiment 23: The method of example embodiment 1 or 11, wherein the therapeutically effective amount of the compound of formula (I) is from about 0.01 mg / kg to about 100 mg / kg of body weight per day.

[0396] Example embodiment 24: The method of example embodiment 1 or 11, wherein the compound of formula (I) is administered once daily, twice daily, three times daily, or four times daily.

[0397] Example embodiment 25: The method of example embodiment 1 or 11, wherein the compound of formula (I) is administered orally, parenterally, subcutaneously, intravenously, intramuscularly, or topically.

[0398] During this prophetic work, in some embodiments of Example embodiment 4 and Example embodiment 18, the AA is selected from basic amino acids and analogs as defined for the main aspects, while in other embodiments AA may be selected from non- basic amino acids, which can modulate properties such as solubility, transport, or tissue distribution without necessarily contributing directly to proton shuttling. In a further nonlimiting example, compounds of formula (I) are evaluated in an animal model of chronic inflammation or autoimmunity to assess effects on tissue pH, inflammatory cytokines, and clinical disease scores. In some embodiments, a collagen-induced arthritis model in mice is employed. Susceptible mice receive type II collagen in complete Freund’s adjuvant according to standard protocols until joint swelling and clinical arthritis are established. Animals are then randomized to receive vehicle or a compound of formula (I) at, for example, about 0.3 mg / kg, about 3 mg / kg, or about 30 mg / kg by oral or parenteral administration once daily for 21-28 days. According to some aspects, endpoints include clinical arthritis scores, paw thickness measurements, body weight, and histopathologic evaluation of joint sections for synovial hyperplasia, inflammatory cell infiltration, cartilage erosion, and bone destruction. Local extracellular pH in affected joints can be assessed using pH-sensitive imaging agents or microelectrodes, and systemic inflammation can be monitored by measuring circulating cytokines such as TNFa, I L-1 p, and IL-6. In some embodiments, treatment with compounds of formula (I) results in partial normalization of joint pH toward physiological values, reduction of inflammatory cytokine levels, and amelioration of clinical and histological signs of arthritis relative to vehicle controls, thereby providing in vivo support for the use of G-L-NH-CO-AA conjugates in chronic inflammatory and autoimmune disorders associated with acidic microenvironments.REFERENCES:1Isselbacher, etal. (1996). HARRISON’S PRINCIPLES OF INTERNAL MEDICINE, 13 ed., 1814-1882.2THE MERCK MANUAL OF DIAGNOSIS AND THERAPY, (2011). 19thEdition, published by Merck Sharp &Dohme Corp., (ISBN 978-0-911910-19-3).Docket No.: 411037-501001 WO THE ENCYCLOPEDIA OF MOLECULAR CELL BIOLOGY AND MOLECULAR MEDICINE, Robert S. Porter et al.(eds.), published by Blackwell Science Ltd., 1999-2012 (ISBN 9783527600908). MOLECULAR BIOLOGY AND BIOTECHNOLOGY: A COMPREHENSIVE DESK REFERENCE, (1995). Robert A.Meyers (ed.), published by VCH Publishers, Inc. (ISBN 1-56081-569-8). IMMUNOLOGY, (2006). Werner Luttmann, published by Elsevier. JANEWAY'S IMMUNOBIOLOGY, (2014). Kenneth Murphy, Allan Mowat, Casey Weaver (eds.), Taylor &Francis Limited, (ISBN 0815345305, 9780815345305). LEWIN'S GENES XI, (2014). published by Jones & Bartlett Publishers (ISBN-1449659055). Michael Richard Green and Joseph Sambrook, (2012). MOLECULAR CLONING: A LABORATORYMANUAL, 4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., USA (ISBN 1936113414). Davis et al., (2012). BASIC METHODS IN MOLECULAR BIOLOGY, Elsevier Science Publishing, Inc., NewYork, USA (ISBN 044460149X). LABORATORY METHODS IN ENZYMOLOGY: DNA, (2013). Jon Lorsch (ed.) Elsevier (ISBN 0124199542). CURRENT PROTOCOLS IN MOLECULAR BIOLOGY (CPMB), (2014). Frederick M. Ausubel (ed.), JohnWiley and Sons (ISBN 047150338X, 9780471503385). CURRENT PROTOCOLS IN PROTEIN SCIENCE (CPPS), (2005). John E. Coligan (ed.), John Wiley andSons, Inc. CURRENT PROTOCOLS IN IMMUNOLOGY (CPI) (2003). John E. Coligan, ADA M Kruisbeek, David HMargulies, Ethan M Shevach, Warren Strobe, (eds.) John Wiley and Sons, Inc. (ISBN 0471142735, 9780471142737).Docket No.: 411037-501001 WO

[0399] All patents and other publications; including literature references, issued patents, published patent applications, and co-pending patent applications; cited throughout this application are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, the methodologies described in such publications that might be used in connection with the technology described herein. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.

[0400] The foregoing written specification is considered to be sufficient to enable one skilled in the art to practice the present aspects and embodiments. The present aspects and embodiments are not to be limited in scope by examples provided, since the examples are intended as a single illustration of one aspect and other functionally equivalent embodiments are within the scope of the disclosure. Various modifications in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims. The advantages and objects described herein are not necessarily encompassed by each embodiment. Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments described herein. Such equivalents are intended to be encompassed by the following claims. It is to be understood that the subject matter of any dependent claim may be combined with the subject matter of any other dependent claim or independent claim, unless such combination is technically incompatible.

Claims

Docket No.: 411037-501001 WOCLAIMSWe clainr.

1. A method of treating a disease in a subject in need thereof, the disease being selected from cancer, cancer-associated inflammation, chronic inflammatory disorders, autoimmune diseases and age-associated systemic inflammation, and combinations thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I):G-L-NH-CO-AA (Formula I),(L-glucose), wherein:G is an L-glucose C-glycosyl residue or a D-glucose C-glycosyl residue in the pyranose form having the carbon numbering shown for formula (I), said residue having a sixmembered ring with a -CH2OH substituent at C-5 and hydroxyl groups at C-2, C-3, C-4 and C-6, each of the hydroxyl groups at C-2, C-3, C-4 and C-6 being independently present as an unsubstituted hydroxyl group or as a group OR7, SR7, NR8R9, O-C(=O)R10or O- C(=O)OR10, and the anomeric carbon C-1 being bonded through a carbon-carbon bond B1 to a carbon atom of the linker group L;Docket No.: 411037-501001 WOR7is hydrogen, Ci-C4alkyl, C2-C6alkenyl, C2-C6alkynyl, benzyl, phenyl or phenyl substituted by one or more substituents selected from Ci-C4alkyl, Ci-C4alkoxy, halogen, hydroxy and nitro;R8and R9are independently hydrogen or Ci-C4alkyl;R10is Ci- C6alkyl, C2-C6alkenyl, C2-C6alkynyl, benzyl or phenyl;L is a divalent linker group connecting the anomeric carbon (C-1) of G to the nitrogen atom of the -NH-CO- group and is selected from Ci-C6straight or branched alkylene and C2-C6alkylene optionally interrupted by one or more heteroatoms selected from O, S and NRa, wherein L contains from 1 to 8 carbon atoms in total and Rais hydrogen or Ci-C4alkyl;AA is the residue of a basic amino acid of formula -NH-CHR11-(CH2)P-NR2R3-CO-, which residue is amide-bonded through its a-carboxyl group to the -NH-CO- moiety of formula (I), wherein p is an integer from 2 to 5;R11is hydrogen, Ci-C6alkyl optionally substituted by one or more substituents selected from hydroxy, amino, carboxy, Ci-C4alkoxycarbonyl and halogen, or is -(CH2)m- NR4R5where m is 1-4 and R4and R5are independently hydrogen or Ci-C4alkyl;R2and R3are independently hydrogen, Ci-C4alkyl, hydroxy-Ci-C4alkyl or a group -C(=NH)NH2, or R2and R3together with the nitrogen atom to which they are attached form a saturated 5-, 6- or 7-membered heterocycle optionally containing an additional nitrogen atom; with the proviso that AA is a residue of a basic amino acid selected from lysine, arginine, ornithine, homoarginine, histidine, diaminobutyric acid and diaminopropionic acid, or a non-naturally occurring analog thereof having a side-chain pKaof at least 7; and wherein the method includes administration of any enantiomer, diastereomer, racemate, pharmaceutically acceptable salt, solvate, hydrate or prodrug of the compound of formula (I), thereby modulating an abnormal intracellular-to-extracellular pH gradient in diseased cells in the subject.

2. The method of claim 1 , wherein the disease is cancer.

3. The method of claim 2, wherein the cancer is a solid tumor selected from breast cancer, lung cancer, colorectal cancer, pancreatic cancer, prostate cancer, ovarian cancer and glioblastoma.

4. The method of claim 1 , wherein the disease is a chronic inflammatory or autoimmune condition selected from rheumatoid arthritis, inflammatory bowel disease, Crohn’s disease,Docket No.: 411037-501001 WO ulcerative colitis, psoriasis, psoriatic arthritis, systemic lupus erythematosus and multiple sclerosis.

5. The method of any one of claims 1-4, wherein administration of the compound of formula (I) increases extracellular pH and / or decreases intracellular pH in diseased cells so that the intracellular-to-extracellular pH gradient approaches that of corresponding non- malignant cells.

6. The method of any one of claims 1-5, wherein the modulation of the pH gradient induces apoptosis in diseased cells, as evidenced by activation of caspase-3 / 7 and loss of clonogenic survival.

7. The method of any one of claims 1-6, wherein restoration of a physiological pH gradient enhances anti-tumor immune surveillance, including increased infiltration and cytotoxic activity of CD8+T cells and / or natural killer cells.

8. The method of any one of claims 1-7, wherein the compound of formula (I) is administered to the subject orally, intravenously, intraperitoneally, intratumorally, peri- tumorally or by a combination of such routes.

9. The method of any one of claims 1-8, wherein the compound of formula (I) is administered at a dose in the range of about 0.01 to about 100 mg / kg body weight per day.

10. The method of any one of claims 1-9, further comprising administering to the subject at least one additional therapeutic agent selected from a cytotoxic chemotherapeutic agent, a targeted small-molecule kinase inhibitor, an immune checkpoint inhibitor, a PARP inhibitor, a hormone or endocrine therapy agent, a monoclonal antibody or antibody-drug conjugate, an epigenetic agent or modulator, and a cell-based immunotherapy.

11. The method of any one of claims 1-10, further comprising co-administering to the subject at least one reverse transcriptase inhibitor (RTI), optionally a nucleoside or nucleotide RTI selected from lamivudine, censavudine and elvucitabine.

12. The method of any one of claims 1-11, wherein L is -CH2-CH2-13. The method of any one of claims 1-12, wherein AA is in the L-configuration at the a- carbon and is the residue of lysine or arginine.Docket No.: 411037-501001 WO14. The method of any one of claims 1-13, wherein the compound of formula (I) is a compound selected from the group consisting of [structure A] and [structure B]:, [structure B]; wherein R is selected from H, Ci-6alkyl, Ci-6hydroxyalkyl, Ci-4fluoroalkyl, C3-6cycloalkyl, benzyl, phenethyl, phenyl, substituted phenyl, heteroaryl, and / or pyridyl, thienyl, or furyl.

15. The method of any one of claims 1-14, wherein at least one of the hydroxyl groups at C-2, C-3, C-4 and C-6 is acylated as an O-C(=O)R10or O-C(=O)OR10group to provide a lipophilically masked prodrug.

16. The method of any one of claims 1-15, for reducing the risk of tumor recurrence or metastatic spread in a subject who has previously received a primary cancer therapy comprising surgery, chemotherapy, radiotherapy, immunotherapy or a combination thereof.

17. A compound of formula (I): G-L-NH-CO-AA as defined in claim 1, for use in therapy.Docket No.: 411037-501001 WO18. The compound for use according to claim 17, for use in treating cancer in a subject in need thereof.

19. The compound for use according to claim 18, for use in treating a cancer characterized by extracellular acidification and intracellular alkalinization relative to corresponding non-malignant cells.

20. The compound for use according to any one of claims 17-19, for use in treating cancer-associated inflammation in a subject having a solid tumor.

21. The compound for use according to any one of claims 17-20, for use in treating a chronic inflammatory or autoimmune disease characterized by an acidic extracellular microenvironment at a site of inflammation.

22. The compound for use according to any one of claims 17-21, for use in enhancing anti-tumor immune surveillance by normalizing extracellular pH in a tumor microenvironment.

23. The compound for use according to any one of claims 17-22, for use in preventing or reducing metastatic dissemination of a solid tumor.

24. The compound for use according to any one of claims 17-23, for use in preventing tumor recurrence following a primary cancer therapy.

25. The compound for use according to any one of claims 17-24, for use in combination therapy with at least one immune checkpoint inhibitor selected from anti-PD-1, anti-PD-L1 and anti-CTLA-4 antibodies.

26. The compound for use according to any one of claims 17-25, for use in combination therapy with at least one reverse transcriptase inhibitor (RTI) to treat age-associated inflammation or cancer-associated inflammation.

27. The compound for use according to any one of claims 17-26, for use in combination with radiotherapy, wherein the compound of formula (I) is administered so as to reverse an abnormal intracellular-to-extracellular pH gradient and improve the response of the cancer to ionizing radiation.Docket No.: 411037-501001 WO28. A pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I): G-L-NH-CO-AA as defined in claim 1 , and at least one pharmaceutically acceptable carrier, diluent or excipient.

29. The pharmaceutical composition of claim 28, formulated for oral administration and in a solid dosage form selected from a tablet, capsule, granule and powder.

30. The pharmaceutical composition of claim 28, formulated for intravenous administration and in a dosage form selected from a sterile aqueous solution, a sterile aqueous suspension and a lyophilized powder for reconstitution.

31. The pharmaceutical composition of claim 28, formulated for local administration to a tumor or tumor bed and in a dosage form selected from an injectable solution, an injectable suspension, a gel, an in s / tu-forming depot, an implant and a biodegradable polymer matrix.

32. The pharmaceutical composition of any one of claims 28-31 , wherein the compound of formula (I) is present as a pharmaceutically acceptable salt, solvate, hydrate or prodrug.

33. The pharmaceutical composition of any one of claims 28-32, further comprising at least one additional therapeutic agent selected from the additional therapeutic agents recited in claim 10.

34. The pharmaceutical composition of claim 33, wherein the additional therapeutic agent is a cytotoxic chemotherapeutic agent selected from doxorubicin, paclitaxel, carboplatin and combinations thereof.

35. The pharmaceutical composition of any one of claims 28-34, further comprising at least one immune checkpoint inhibitor selected from nivolumab, pembrolizumab, atezolizumab and ipilimumab.

36. The pharmaceutical composition of any one of claims 28-35, further comprising at least one reverse transcriptase inhibitor selected from lamivudine, censavudine and elvucitabine.Docket No.: 411037-501001 WO37. The pharmaceutical composition of any one of claims 28-36, wherein the composition is in unit dosage form containing from about 1 mg to about 500 mg of the compound of formula (I).

38. The pharmaceutical composition of any one of claims 28-37, wherein the composition is formulated as a controlled-release or sustained-release dosage form adapted to provide plasma levels of the compound of formula (I) sufficient to modulate a cancer-associated pH gradient over a period of at least 8 hours.

39. A kit comprising (a) a pharmaceutical composition according to any one of claims 28-38 and (b) printed instructions for use of the composition in the treatment of a disease as defined in claim 1.

40. A pharmaceutical combination comprising:(a) a compound of formula (I): G-L-NH-CO-AA as defined in claim 1 ; and(b) at least one reverse transcriptase inhibitor (RTI); wherein components (a) and (b) are formulated or packaged for concurrent, sequential or separate administration to a subject in need thereof.

41. The pharmaceutical combination of claim 40, wherein the at least one RTI is a nucleoside or nucleotide RTI.

42. The pharmaceutical combination of claim 41 , wherein the at least one RTI is selected from lamivudine, emtricitabine, tenofovir, abacavir, zidovudine and combinations thereof.

43. The pharmaceutical combination of any one of claims 40-42, wherein the at least one RTI is selected from censavudine, elvucitabine and combinations thereof.

44. The pharmaceutical combination of any one of claims 40-43, further comprising at least one additional therapeutic agent selected from a cytotoxic chemotherapeutic agent, a targeted kinase inhibitor and an immune checkpoint inhibitor.

45. The pharmaceutical combination of any one of claims 40-44, for use in treating cancer or a cancer-associated inflammatory disorder in a subject in need thereof.Docket No.: 411037-501001 WO46. The pharmaceutical combination of any one of claims 40-44, for use in treating an age-associated or chronic inflammatory condition characterized by an acidic microenvironment.

47. The pharmaceutical combination of any one of claims 40-46, wherein the compound of formula (I) is administered prior to or concurrently with the at least one RTI so as to normalize extracellular pH and reduce RTI-sensitive inflammatory signaling within a tumor microenvironment.

48. The pharmaceutical combination of any one of claims 40-47, wherein the at least one RTI is administered at a daily dose that is less than or equal to about 80% of a standard adult monotherapy dose for treatment of viral infection, while maintaining a therapeutically effective level for modulation of age- or cancer-associated inflammation.

49. The pharmaceutical combination of any one of claims 40-48, wherein component (a) is formulated for oral administration and component (b) is formulated for oral or parenteral administration.

50. A kit comprising (i) a first container containing a compound of formula (I) as defined in claim 1 or a pharmaceutical composition according to any one of claims 28-38, (ii) a second container containing at least one reverse transcriptase inhibitor as defined in any one of claims 41-43, and (iii) printed instructions for administration of the contents of the first and second containers as combination therapy to treat a disease as defined in claim 1.

51. A method of selectively killing cancer cells in vitro, the method comprising contacting a population of cancer cells with an effective amount of a compound of formula (I) as defined in claim 1 under conditions sufficient to reverse an abnormal intracellular-to-extracellular pH gradient in the cancer cells and thereby induce apoptosis in the cancer cells.

52. The method of claim 51 , wherein the cancer cells comprise breast cancer cells, optionally estrogen receptor-positive MCF-7 cells or MCF-7-like cells and are cultured as monolayers or as multicellular spheroids.

53. The method of claim 51 or 52, further comprising measuring intracellular and / or extracellular pH before and after contacting the cells with the compound of formula (I) using a pH-sensitive fluorescent indicator.Docket No.: 411037-501001 WO54. A method for assessing the sensitivity of tumor cells from a subject to a compound of formula (I), the method comprising:(a) obtaining tumor cells from the subject;(b) contacting the tumor cells ex vivo with at least one test concentration of a compound of formula (I) as defined in claim 1; and(c) determining at least one response selected from a change in cell viability, a change in apoptosis and a change in intracellular or extracellular pH; wherein the magnitude of the response in step (c) is used to assess sensitivity of the tumor cells to the compound of formula (I).

55. The method of claim 54, further comprising, on the basis of the assessed sensitivity, selecting a treatment regimen for the subject that comprises administration of a therapeutically effective amount of a compound of formula (I).

56. The method of any one of claims 1-16, wherein the diseased cells exhibit an extracellular pH prior to treatment in the range of about 6.0 to about 7.0 and treatment with the compound of formula (I) increases the extracellular pH in the diseased tissue to a value in the range of about 7.0 to about 7.5.

57. A kit for assessing sensitivity of cancer cells to a compound of formula (I), the kit comprising:(a) a compound of formula (I) as defined in claim 1 ;(b) a pH-sensitive fluorescent indicator suitable for measuring intracellular and / or extracellular pH in living cells; and(c) printed instructions describing a protocol for contacting cancer cells with the compound of formula (I), detecting pH changes using the pH-sensitive fluorescent indicator, and determining sensitivity of the cancer cells to the compound of formula (I) based on the detected changes.

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