Methods of treating retinal diseases

Abstract

Provided herein are methods of using Compound 1, or a pharmaceutically acceptable form thereof, for treating retinal disease, including in combination with a VEGF / VEGFR signaling pathway inhibitor.

Description

Attorney Docket No. 14168-127-228METHODS OF TREATING RETINAL DISEASES1. CROSS REFERENCE

[0001] This application claims the benefit of priority to U.S. Serial No. 63 / 725,462, filed November 26, 2024, which is incorporated herein by reference in its entirety.2. FIELD

[0002] Provided herein are methods of using a famesyltransferase inhibitor that isCompound 1:or a pharmaceutically acceptable form thereof, for treating, preventing, or managing retinal diseases. Further provided herein are methods of using Compound 1, or a pharmaceutically acceptable form thereof, in combination with a vascular endothelial growth factor (VEGF) / VEGF receptor (VEGFR) signaling pathway inhibitor, for example with a VEGF receptor (VEGFR) inhibitor or a VEGFA antagonist, for treating, preventing, or managing retinal diseases.3. BACKGROUND

[0003] Blood vessel formation, or angiogenesis, is a complex and coordinated process that is important in normal development and adult physiology. Each step in the process is regulated by a balance of pro- and anti -angiogenic molecules and as such, in settings where there is perturbation of this balance, pathological states occur. For example, in clear cell renal cell carcinoma (ccRCC), 55-70% of patients harbor an inactivating mutation in the von Hippel- Lindau (VHL) gene, which subsequently leads to up-regulation of VEGFA, a critical pro- angiogenic factor (The Cancer Genome Atlas Research Network, Nature 2013, 499, 43-49; Razafinjatovo C. et al., BMC Cancer 2016, 16, 638). In fact, in many tumors and in certain eye diseases, VEGFA expression is aberrantly high, which is collectively due to genetic or epigenetic alterations, tissue hypoxia, metabolic perturbations, or other factors (Apte R.S. et al., Cell 2019,176(6), P1248-1264).

[0004] VEGFA activates VEGFR-mediated signaling in endothelial cells (EC) that contributes to key angiogenic processes, including tip cell basement degradation and migration, stalk cell proliferation, and lumen formation. Consequently, there are several drugs in oncology and ophthalmology that target the VEGF-VEGFR signaling pathway, including VEGF or VEGFA antagonists (e.g., a biologic or monoclonal antibody, such as pegaptanib (Macugen), bevacizumab, ranibizumab, ramucirumab, aflibercept, conbercept, brolucizumab, or faricimab) and tyrosine kinase inhibitors that are VEGFR inhibitors (TKI; e.g., apatinib, axitinib, cabozantinib, lenvatinib, pazopanib, regorafenib, sorafenib, sunitinib, and vandetanib) (Cao Y. et al., Nat. Rev. Drug Discov. 2023, 22, 476-495).

[0005] Farnesyltransferase inhibitors (FTI) have also been reported to have anti angiogenic activity. For example, the FTI tipifamib inhibited sprouting and branching of human umbilical vein endothelial cells (HUVEC) in vitro and treatment of a Harvey rat sarcoma virus protein (HRAS)-mutant head and neck squamous cell carcinoma (HNSCC) patient-derived xenograft (PDX) model with tipifamib led to reduced tumor vascularity, as assessed by CD31 expression (Gilardi M. et al. Mol. Cancer Ther. 2020, 19(9), 1784-1796). Another FTI, lonafarnib, was also shown to cause decreased EC motility, possibly through lonafamib-mediated disturbance of EC polarity or centrosome reorientation (Sun L. et al., PLoS ONE 2015, 10(4), e0122830). As described herein, the anti angiogenic activity of Compound 1, or a pharmaceutically acceptable form thereof, an FTI, is useful in methods of treating retinal diseases, alone or in combination with other therapies, such as VEGF / VEGFR signaling pathway inhibitors.4. SUMMARY

[0006] In one aspect is a method of treating a retinal disease in a subject comprising administering to the subject Compound 1, or a pharmaceutically acceptable form thereof (or a pharmaceutical composition comprising the same).

[0007] In one aspect is a method of treating a retinal disease in a subject comprising administering to the subject Compound 1, or a pharmaceutically acceptable form thereof (or a pharmaceutical composition comprising the same), and a VEGF / VEGFR signaling pathway inhibitor (e g., a VEGFR inhibitor or a VEGFA antagonist).

[0008] In one aspect is a method of inhibiting angiogenesis in a subject, comprising administering Compound 1, or a pharmaceutically acceptable form thereof.

[0009] In one aspect is a method of inhibiting VEGF / VEGFR signaling pathway in a subject, comprising administering Compound 1, or a pharmaceutically acceptable form thereof.

[0010] In one aspect is a method of inhibiting angiogenesis in a subject, comprising administering: (i) Compound 1, or a pharmaceutically acceptable form thereof; and (ii) a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist).

[0011] In one aspect is a method of inhibiting VEGFA / VEGFR signaling pathway in a subject, comprising administering: (i) Compound 1, or a pharmaceutically acceptable form thereof; and (ii) a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist).

[0012] In another aspect, the subject has a retinal disease.

[0013] In another aspect is a pharmaceutical composition comprising Compound 1, or a pharmaceutically acceptable form thereof.

[0014] In another aspect is a pharmaceutical composition comprising: (a) Compound 1, or a pharmaceutically acceptable form thereof; and (b) a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist).

[0015] In another aspect is a pharmaceutical kit comprising Compound 1, or a pharmaceutically acceptable form thereof.

[0016] In another aspect is a pharmaceutical kit comprising: (a) Compound 1, or a pharmaceutically acceptable form thereof; and (b) a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist).

[0017] In another aspect, provided herein is a pharmaceutical packaging comprising: Compound 1, or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient.

[0018] In another aspect, provided herein is a pharmaceutical packaging comprising: (l)(a) Compound 1, or a pharmaceutically acceptable form thereof; and (b) a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist); or (2)(a) a pharmaceutical composition comprising Compound 1, or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient; and (b) a pharmaceutical composition comprising a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist), and a pharmaceutically acceptable excipient. In another aspect is a pharmaceutical composition comprising Compound 1, or a pharmaceutically acceptable form thereof, for use in the methodsdescribed herein.5. BRIEF DESCRIPTION OF THE FIGURES

[0019] FIG. 1 : Plot of tumor volume over time for treatment of A498 RCC CDX with the compound of Formula (I) (i.e., Compound 1), axitinib, or the combination.

[0020] FIG. 2: Plot of tumor volume over time for treatment of KI- 12-0073 RCC PDX with the compound of Formula (I) (i.e., Compound 1), axitinib, or the combination.

[0021] FIGS. 3A-3B: Combination treatment of the compound of Formula (1) (i.e., Compound 1) and cabozantinib inhibited tumor growth in RCC CDX models. FIG. 3A: 786-0 CDX treated with the compound of Formula (I) (i.e., Compound 1) (20 mg / kg, BID) and cabozantinib (20 mg / kg, QD) (lane 3) compared to treatment with the compound of Formula (I) (i.e., Compound 1) (lane 1) or cabozantinib (lane 2) alone; FIG. 3B: A498 CDX treated with the compound of Formula (I) (i.e., Compound 1) and cabozantinib (8 or 20 mg / kg, QD) (lanes 4 and 5, respectively) compared to treatment with the compound of Formula (I) (i.e., Compound 1) alone (lane 1) or cabozantinib alone (lanes 2 and 3).

[0022] FIGS. 4A-4F: Combination of the compound of Formula (I) (i.e., Compound 1) and cabozantinib inhibited tumor growth in RCC PDX and CDX models for the compound of Formula (I) (i.e., Compound 1), cabozantinib, and the combination. Plot of tumor volume over time (FIG. 4A) and plot of % tumor volume change (FIG. 4B) in a KI- 12-0073 VHL-mutant ccRCC PDX model; plot of tumor volume over time (FIG. 4C) and plot of % tumor volume change (FIG. 4D) in a 786-0 CDX model; and plot of tumor volume over time (FIG. 4E) and plot of % tumor volume change (FIG. 4F) in a KI-0326 VHL-mutant ccRCC PDX model.

[0023] FIG. 5 : Plot of % tumor volume change at day 28 relative to day 0 for mice with786-0 VHL-mutant CDX treated with the compound of Formula (I) (i.e., Compound 1) (20 mg / kg, BID) and cabozantinib (4, 8, 10, or 12 mg / kg, QD), alone or in combination.

[0024] FIG. 6 : Plot of tumor volume over time for 786-0 CDX mice treated with cabozantinib, the compound of Formula (1) (i.e., Compound 1), lenvatinib, lenvatinib plus everolimus, the compound of Formula (I) (i.e., Compound 1) plus cabozantinib, and the compound of Formula (I) (i.e., Compound 1) with lenvatinib.

[0025] FIG. 7: Plot of tumor volume over time for 786-0 CDX mice treated with the compound of Formula (I) (i.e., Compound 1), cabozantinib, axitinib, or the combination of the compound of Formula (I) (i.e., Compound 1) and cabozantinib.

[0026] FTG. 8 : Immunoblot of cell signaling markers from 786-0 CDX cells following treatment with cabozantinib, the combination of Formula (I) (i.e., Compound 1), or the combination.

[0027] FIGS. 9A-9C: Plots of % cell viability of HUVEC cells treated with varying concentrations of the compound of Formula (I) (i.e., Compound 1) and varying concentrations of cabozantinib (FIG. 9A), axitinib (FIG. 9B), or lenvantinib (FIG. 9C).

[0028] FIGS. 10A-10F: Tube formation in HUVEC cells was inhibited by the combination of the compound of Formula (I) (i.e., Compound 1) and axitinib or cabozantinib. FIG. 10A: vehicle; FIG. 10B: axitinib; FIG. IOC: cabozantinib; FIG. 10D: compound of Formula (I) (i.e., Compound 1); FIG. 10E: axitinib and compound of Formula (I) (i.e., Compound 1); FIG. 10F: cabozantinib and compound of Formula (I) (i.e., Compound 1).

[0029] FIGS. 11A-11B: GFP imaging (FIG. HA) and plots of number of master segments and total master segments length (FIG. 11B) of the effect on tube formation in GFP-labeled HUVEC cells treated with vehicle, the compound of Formula (I) (i.e., Compound 1), cabozantinib, or the combination.

[0030] FIG. 12: Plot of cell death, as percentage normalized to baseline, over time, in HUVEC cells treated with the compound of Formula (I) (i.e., Compound 1), cabozantinib, or the combination (with staurosporine as control).

[0031] FIGS. 13A-13D: Plot of tumor volume over time for treatment of KI-12-0073 PDX (FIG. 13A), KI-12-0097 PDX (FIG. 13B), Cakil CDX (FIG. 13C), and KI-12-0351 PDX (FIG. 13D) with either cabozantinib or Compound 1.

[0032] FIG. 14: Immunoblot of indicated MAPK / PI3K signaling proteins and cell cycle arrest markers in HUVEC cells stimulated with VEGFA for the indicated timepoints, with or without treatment with cabozantinib, Compound 1, or the combination.6. DETAILED DESCRIPTION

[0033] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art. In the event that there are a plurality of definitions for a term herein, those in this section prevail unless stated otherwise.

[0034] As used herein, and in the specification and the accompanying claims, the indefinite articles “a” and “an” and the definite article “the” include plural as well as single referents unlessthe context clearly indicates otherwise.

[0035] As used herein, and unless otherwise specified, the terms “about” and “approximately,” when used in connection with doses, amounts, or weight percentages of ingredients of a composition or a dosage form, mean a dose, amount, or weight percent within 30%, within 20%, within 15%, within 10%, or within 5%, of the specified dose, amount, or weight percent.

[0036] As used herein, a “pharmaceutically acceptable form” of compounds disclosed herein includes, but is not limited to, a pharmaceutically acceptable salt, solvate, isomer, and isotopologue (i.e., isotopically labeled derivative), of compounds disclosed herein, which includes combinations thereof (e.g., a solvate of a pharmaceutically acceptable salt, or an isomer and / or isotopologue of a compound or of a solvate, salt, or solvate of salt of such compound). In some embodiments, a “pharmaceutically acceptable form” includes, but is not limited to, a pharmaceutically acceptable salt, solvate, isomer e.g., tautomer or stereoisomer), and isotopologue (z.e., isotopically labeled derivative) of Compound 1 as disclosed herein.

[0037] The term “isomer” as used herein comprises a stereoisomer or tautomer as defined herein. As used herein, the term “stereoisomers” is understood to mean isomers that differ only in the way the atoms are arranged in space. As used herein, the term “isomer” includes any and all geometric isomers and stereoisomers. For example, “isomers” include geometric double bond cis- and / ra -isomers, also termed E- and Z- isomers; R- and S-enantiomers; diastereomers, (t / )-i somers and ( / )-isomers, racemic mixtures thereof; and other mixtures thereof, as falling within the scope of this disclosure.

[0038] As used herein and unless otherwise indicated, the term “stereoisomerically pure” means one stereoisomer of a compound that is substantially free of other stereoisomers of that compound. For example, a stereoisomerically pure compound having one chiral center will be substantially free of the opposite enantiomer of the compound. In some embodiments is the stereoisomerically pure Compound 1 (i.e., (5)-3-amino-3-(l -methyl- l / / -imidazol-5-yl)-6-oxa- 2(4,6)-quinolina-l,4(l,3)-dibenzenacyclohexaphane-22,44-dicarbonitrile), substantially free of (A)-3-amino-3-(l-methyl-l / / -imidazol-5-yl)-6-oxa-2(4,6)-quinolina-l,4(l,3)- dibenzenacyclohexaphane-22,44-dicarbonitrile). A stereoisomerically pure compound having two chiral centers will be substantially free of other diastereomers of the compound. A typical stereoisomerically pure compound comprises greater than about 80% by weight of onestereoisomer of the compound and less than about 20% by weight of other stereoisomers of the compound, greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomers of the compound, greater than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of the other stereoisomers of the compound, or greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomers of the compound. The compounds can have chiral centers and can occur as racemates, individual enantiomers or diastereomers, and mixtures thereof. All such isomeric forms are included within the embodiments provided herein, including mixtures thereof.

[0039] It is to be understood that the compounds provided herein may contain chiral centers. Such chiral centers may be of either the (A) or ( ) configuration, or may be a mixture thereof. It is to be understood that the chiral centers of the compounds provided herein may undergo epimerization in vivo. As such, one of skill in the art will recognize that administration of a compound in its (A) form is equivalent, for compounds that undergo epimerization in vivo, to administration of the compound in its (S) form. Optically active (+) and (-), (7?)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as chromatography on a chiral stationary phase.

[0040] The use of stereoisomerically pure forms of such compounds, as well as the use of mixtures of those forms, are encompassed by the embodiments provided herein. For example, mixtures comprising equal or unequal amounts of the enantiomers of a particular compound may be used in methods and compositions provided herein.

[0041] These isomers may be asymmetrically synthesized or resolved using standard techniques such as chiral columns or chiral resolving agents. See, e.g., Jacques, J., et al., (Wiley-Interscience, New York, 1981); Wilen, S. H., et al., Tetrahedron 33:2725 (1977); Eliel, E. L., Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, S. H., Tables of Resolving Agents and Optical Resolutions p. 268 (E.L. Eliel, Ed., Univ, of Notre Dame Press, Notre Dame, IN, 1972); Todd, M., Separation Of Enantiomers : Synthetic Methods (Wiley -VCH Verlag gmbH & Co. KGaA, Weinheim, Germany, 2014); Toda, F., Enantiomer Separation: Fundamentals and Practical Methods (Springer Science & Business Media, 2007);Subramanian, G. Chiral Separation Techniques: A Practical Approach (John Wiley & Sons, 2008); Ahuja, S., Chiral Separation Methods for Pharmaceutical and Biotechnological Products(John Wiley & Sons, 2011). Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as chromatography on a chiral stationary phase.

[0042] In certain embodiments, the pharmaceutically acceptable form is an atropisomer. Atropisomers are stereoisomers resulting from hindered rotation about a single bond axis where the rotational barrier is sufficient to allow for isolation of the rotational isomers.

[0043] In certain embodiments, the pharmaceutically acceptable form is a tautomer. As used herein, the term “tautomer” is a type of isomer that includes two or more interconvertable compounds resulting from at least one formal migration of a hydrogen atom and at least one change in valency (e.g., a single bond to a double bond, a triple bond to a double bond, or a triple bond to a single bond, or vice versa). “Tautomerization” includes prototropic or protonshift tautomerization, which is considered a subset of acid base chemistry. “Prototropic tautomerization” or “proton-shift tautomerization” involves the migration of a proton accompanied by changes in bond order. The exact ratio of the tautomers depends on several factors, including temperature, solvent, and pH. Where tautomerization is possible (e.g., in solution), a chemical equilibrium of tautomers can be reached. Tautomerizations (i.e.. the reaction providing a tautomeric pair) can be catalyzed by acid or base, or can occur without the action or presence of an external agent. The concentrations of the isomeric forms will depend on the environment the compound is found in and may be different depending upon, for example, whether the compound is a solid or is in an organic or aqueous solution. Exemplary tautomerizations include, but are not limited to, keto-enol; amide-imide; lactam-lactim; enamineimine; and enamine-(a different) enamine tautomerizations. For example, in aqueous solution, pyrazoles may exhibit the following isomeric forms, which are referred to as tautomers of each other:

[0044] As readily understood by one skilled in the art, a wide variety of functional groups and other structures may exhibit tautomerism and all tautomers of a compound are within the scope of the compound as provided herein.

[0045] In certain embodiments, a compound described herein is in the form of a pharmaceutically acceptable salt. As used herein, the term “pharmaceutically acceptable salt”refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of subjects without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail (see J. Pharm. Sci. (1977) 66: 1-19). Pharmaceutically acceptable salts of the compounds provided herein include those derived from suitable inorganic and organic acids and bases, such as suitable inorganic and organic addition acids and bases.

[0046] In certain embodiments, the pharmaceutically acceptable form of a compound disclosed herein is exclusive of a salt form (i.e., is not a salt), sometimes referred to as a free form or free base form, of a compound disclosed herein. In some embodiments are solvates of such free base forms.

[0047] In certain embodiments, a compound as described herein is in the form of a solvate (e.g., a hydrate). As used herein, the term “solvate” refers to a compound that further includes a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. The solvate can be of a disclosed compound or a pharmaceutically acceptable salt thereof. Where the solvent is water, the solvate is a “hydrate.” In some embodiments, the solvate is a hydrate. Pharmaceutically acceptable solvates and hydrates are complexes that, for example, can include 0.1, 0.25, 0.50, 0.75, or 1 solvent or water molecules, or can include 1 to about 100, or 1 to about 10, or one to about 2, about 3 or about 4, solvent or water molecules. It will be understood that the term “compound” as used herein encompasses the compound (or a pharmaceutically acceptable salt thereof) and solvates of the compound or of pharmaceutically acceptable salts thereof, as well as mixtures thereof.

[0048] The term “isotopologue” refers to isotopically-enriched compounds that are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Unless otherwise stated, structures depicted herein are also meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. Examples of isotopes that can be incorporated into compounds described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as2H,3H,13C,14C,15N,17O,180,32P,33P,33S,34S,35S,36S,18F,35C1,36C1, and37C1, respectively, each of which is also within the scope of this description. For example, compounds having the present structures except forthe replacement or enrichment of a hydrogen by deuterium or tritium at one or more atoms in the molecule, are within the scope of this disclosure. In one embodiment, provided herein are isotopically labeled compounds having one or more hydrogen atoms replaced by or enriched by deuterium. When the compounds are enriched with deuterium, the deuterium-to-hydrogen ratio on the deuterated atoms of the molecule substantially exceeds the naturally occurring deuterium- to-hydrogen ratio. In one embodiment, provided herein are isotopically labeled compounds having one or more hydrogen atoms replaced by or enriched by tritium. Further, substitution with heavier isotopes such as deuterium (i.e.,2H) can afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). Isotopically labeled compounds disclosed herein can generally be prepared by substituting an isotopically labeled reagent for a non-isotopically labeled reagent. Isotopically- enriched compounds can generally be prepared using procedures known to persons of ordinary skill in the art by substituting an appropriate isotopically-enriched reagent for a non-isotopically- enriched reagent. An embodiment described herein may include an isotopologue form wherein the isotopologue is substituted on one or more atom members of said compound with one or more deuterium atoms in place of one or more hydrogen atoms. An embodiment described herein may include a compound wherein a carbon atom may have from 1 to 3 hydrogen atoms optionally replaced with deuterium.

[0049] As used herein, compounds disclosed herein include, but are not limited to, free base forms or pharmaceutically acceptable salts thereof, and solvates or hydrates thereof, and isotopologues (i.e., isotopically labeled derivative) of such compounds. In some embodiments are contemplated a free base or pharmaceutically acceptable salt of Compound 1, or a hydrate or solvate and / or isotopologue (i.e., isotopically labeled derivative) thereof, or analogous forms of a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist).

[0050] It should be noted that if there is a discrepancy between a depicted structure and a name for that structure, the depicted structure is to be accorded more weight.

[0051] As used herein, the term “pharmaceutically acceptable excipient” means a carrier, vehicle, compendial ingredient, diluent, or other additive approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans. The term “excipient” refers to a diluent, adjuvant (e.g., Freund’s adjuvant (complete and incomplete)), additive, or vehiclewith which a therapeutic agent is administered. Such pharmaceutical excipients can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Water is a specific vehicle for intravenously administered pharmaceutical compositions. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid vehicles, particularly for injectable solutions. For example, pharmaceutically acceptable excipients include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions as disclosed herein is contemplated. Supplementary active ingredients can also be incorporated into the pharmaceutical compositions. Examples of excipients that can be used in oral dosage forms provided herein include, but are not limited to, binders, fdlers, disintegrants, and lubricants.

[0052] As used herein, a “VEGF / VEGFR signaling pathway inhibitor” is an agent that inhibits VEGF signaling. In some aspects, a VEGF / VEGFR signaling pathway inhibitor is a VEGFR inhibitor or a VEGF A antagonist. In other aspects, a VEGF / VEGFR signaling pathway inhibitor is a VEGFC inhibitor or a VEGFD inhibitor.

[0053] As used herein, a “VEGF A antagonist” is an agent that antagonizes the VEGFR / VEGFR2 axis. Non-limiting examples include agents that function by neutralization of VEGFA (e.g., a monoclonal antibody such as bevacizumab), that operate as decoy receptor traps (which sequester circulating VEGF to prevent downstream receptor activation, such as aflibercept), or direct anti-VEGFR2 agents (e.g., a monoclonal antibody such as ramucirumab). In some aspects, the VEGFA antagonist is a biologic or monoclonal antibody, such as pegaptanib (Macugen), bevacizumab, ranibizumab, ramucirumab, aflibercept, conbercept, brolucizumab, or faricimab.

[0054] As used herein, the term “VEGFR inhibitor” means a small molecule compound that inhibits one or more VEGFR isoforms (e.g., VEGFR-1, VEGFR-2, VEGFR-3) with an IC50 value of less than or equal to 500 nM in a biochemical or cellular assay. A VEGFR inhibitor may inhibit one or more VEGFR isoforms and also other tyrosine kinase targets, such as FGFR-1, -2, -3, or -4, PDGFR-a or -0, KIT, RET, MET, AXL, ROS1, TYRO3, MER, TRKB, FLT-3, TIE-2, DDR2, TRKA, EPH2A, RAF-1, BRAF, BRAF V600E, SAPK2, PTK5, ABL, orCSF-1R, or a combination thereof. In some embodiments, a VEGFR inhibitor inhibits at least one VEGFR isoform and at least one of PDGFR-a and PDGFR-p. In some embodiments, a VEGFR inhibitor is a Type 1 kinase inhibitor, or a Type II inhibitor, or a covalent inhibitor. Exemplary VEGFR inhibitors include, but are not limited to, cabozantinib, lenvantinib, axitinib, regorafenib, vandetanib, pazopanib, sunitinib, sorafenib, tivozanib, fruquintinib, vorolanib, and zanzalintinib, and pharmaceutically acceptable forms thereof. In some embodiments, the VEGFR inhibitor is cabozantinib, lenvantinib, axitinib, pazopanib, sunitinib, sorafenib, or tivozanib, or a pharmaceutically acceptable form thereof. In some embodiments, the VEGFR inhibitor is zanzalintinib, or a pharmaceutically acceptable form thereof. In some embodiments, the VEGFR inhibitor is fruquintinib, or a pharmaceutically acceptable form thereof. In some embodiments, the VEGFR inhibitor is cabozantinib, axitinib, sunitinib, or sorafenib, or a pharmaceutically acceptable form thereof. In some embodiments, the VEGFR inhibitor is axitinib or vorolanib, or a pharmaceutically acceptable form thereof. In some embodiments the VEGFR inhibitor is cabozantinib, or a pharmaceutically acceptable form thereof. In some embodiments, the pharmaceutically acceptable form of the VEGFR inhibitor is cabozantinib (S)- malate, lenvantinib mesylate, axitinib free base, regorafenib monohydrate, vandetanib free base, pazopanib hydrochloride, sunitinib (S)-malate, sorafenib tosylate, tivozanib hydrochloride hydrate, fruquintinib free base, or zanzalintinib fumarate.

[0055] As used herein, the terms “prevention” and “preventing” refer to obtaining beneficial or desired results including, but not limited, to prophylactic benefit. For prophylactic benefit, the compounds and pharmaceutical compositions disclosed herein can be administered according to the methods of treating as provided herein to a patient at risk of developing a retinal disease, to a patient reporting one or more of the physiological symptoms of a retinal disease, even though a diagnosis of the retinal disease may not have been made.

[0056] As used herein and unless otherwise indicated, the term “effective amount” in connection with a compound means an amount capable of treating, preventing, or managing a disorder, disease or condition, or symptoms thereof. In some embodiments, an effective amount of Compound 1 or a pharmaceutically acceptable form thereof, an effective amount of VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist) and / or an effective amount in the context of a combination thereof, can provide one or more benefits according to the methods of treating provided herein. For example, the effective amountof Compound 1 or a pharmaceutically acceptable form thereof, the effective amount of a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist), and / or the effective amount in the context of a combination thereof, can prevent, treat, and / or ameliorate one or more symptoms associated with a retinal disease.

[0057] In some embodiments, where a dose amount, a per day dose amount, or an amount in a pharmaceutical composition, pharmaceutical kit, or pharmaceutical packaging is described for a compound that is in a pharmaceutically acceptable salt and / or solvate form, the amount is expressed as the mass of the compound in its free form (e.g., free base) equivalent amount (z.c., the form of the compound exclusive of the salt and unsolvated). The amount is referred to as “free form equivalent” or “free base equivalent.”

[0058] As used herein, the terms “continuous dosing” and “continuous dosing schedule,” or “continuous” and “continuously” in the context of administering, refer to daily administration, such as once daily (QD), twice daily (BID), three times daily (TID), or four times a day (QID), of Compound 1 or a pharmaceutically acceptable form thereof, or of the VEGF / VEGFR signaling pathway inhibitor (e.g., VEGFR inhibitor or VEGFA antagonist), or a combination thereof, as disclosed herein. In the context of infusion or port delivery dosing, “continuous” dosing refers to infusion of the agent over a period of time.

[0059] A “treatment cycle” as understood herein is a given period of time during which one or more treatments are administered to a subject in need thereof. In some embodiments, a treatment cycle is a 28-day treatment cycle.

[0060] As used herein and unless otherwise indicated, the term “subject” to which administration is contemplated, can be an animal, including, but not limited to, a human (e.g., a male or female of any age group, such as an adult subject or an adolescent subject); primates (e.g., cynomolgus monkeys, rhesus monkeys), and / or other mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, dogs, rabbits, rodents, and / or birds (e.g., commercially relevant birds such as chickens, ducks, geese, quail, and / or turkeys). In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the subject is an adolescent human. In some embodiments, the subject is an adult human. In some embodiments, the subject is a patient, for example, a human patient.

[0061] As used herein, the term “retinal disease” refers to a disorder that affects or causes damage to the retina. Retinal diseases can lead to sever vision loss or blindness. In someembodiments, the subject has, suffers from, has symptoms associated with, or is diagnosed as having, a retinal disease. In some embodiments, the subject has or suffers from a retinal disease. In some embodiments, the subject has symptoms associated with a retinal disease. In some embodiments, the subject is diagnosed as having a retinal disease. In some embodiments, the subject may be diagnosed as having a retinal disease by one skilled in the art, for example, a physician, such as an ophthalmologist.

[0062] The term “diabetic retinopathy” refers to a chronic retinal disease that occurs when high blood sugar caused by diabetes damages the blood vessels in the retina.

[0063] The term “diabetic macular edema” (DME) refers to a diabetic retinopathy causing swelling in the macula.

[0064] The term “age-related macular degeneration” (AMD) refers to a chronic retinal disease occurring when aging causes damage to the macula. AMD can be atrophic AMD (also known as dry AMD), when the light-sensitive cells of the macula break down slowly, resulting in a thinner macula. AMD can be advanced neovascular AMD (also known as wet AMD), when abnormal blood vessels grow in the back of the eye and damage the macula.

[0065] The term “retinal vein occlusion” (RVO) refers to a condition occurring when a vein draining blood from the retina is blocked.

[0066] The term “retinopathy of prematurity” refers to a retinal disease occurring when abnormal blood vessels grow in the retina of a premature baby, or in the retina of a baby weighing less than 3 pounds.

[0067] In some embodiments, the subject is a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or VEGFA antagonist)-naive subject. In some embodiments, the subject is a subject naive to treatment with a VEGFR inhibitor, such as one or more of cabozantinib, lenvantinib, axitinib, regorafenib, vandetanib, pazopanib, sunitinib, sorafenib, tivozanib, fruquintinib, vorolanib, and zanzalintinib, and pharmaceutically acceptable forms thereof. In some embodiments, the subject has been treated previously with a VEGF / VEGFR signaling pathway inhibitor. For example, in some aspects, the subject has been treated previously with a VEGFR inhibitor and is not currently being treated with a VEGFR inhibitor, or the subject is currently being treated with the same or a different VEGFR inhibitor, optionally with the same VEGFR inhibitor, and Compound 1, or a pharmaceutically acceptable form thereof, is added to the existing VEGFR inhibitor regimen. In some embodiments, the subjecthas been treated previously with a VEGFA antagonist (e.g., a biologic or monoclonal antibody, such as pegaptanib (Macugen), bevacizumab, ranibizumab, ramucirumab, aflibercept, conbercept, brolucizumab, or faricimab) and is not currently being treated with the VEGFA antagonist, or such treatment is ongoing and the Compound 1 regimen is added.

[0068] As used herein and unless otherwise indicated, the terms “treat,” “treating,” “treatment,” and “ameliorating” are used interchangeably herein, and means an alleviation, in whole or in part, of a disorder, disease or condition, such as a retinal disease, or one or more of the symptoms associated with a disorder, disease, or condition, such as a retinal disease, or slowing or halting of further progression or worsening of those symptoms, or alleviating or eradicating the cause(s) of the disorder, disease, or condition itself, such as a retinal disease. In some embodiments, these terms refer to an approach for obtaining beneficial or desired results including, but not limited to, a therapeutic benefit or a prophylactic benefit. A therapeutic benefit resulting from the methods of treatment provided herein includes the eradication or amelioration of the underlying disorder, such as a retinal disease, being treated, the eradication or amelioration of one or more of the physiological signs or symptoms associated with the underlying disorder (e.g., a retinal disease) such that an improvement is observed in the patient, notwithstanding that the patient can still be afflicted with the underlying disease or disorder (e.g., a retinal disease). For example, when used in reference to a patient having a retinal disease, therapeutic benefit refers to an action that reduces the severity of the a retinal disease, or retards or slows the progression of the a retinal disease. A prophylactic benefit resulting from the methods of treatment provided herein includes delaying or eliminating the appearance of a disease or disorder (e.g., a retinal disease), delaying or eliminating the onset of symptoms of a disease or disorder (e.g., a retinal disease), slowing, halting, or reversing the progression of a disease or disorder (e.g., a retinal disease), or any combination thereof.

[0069] In some embodiments, the methods described herein improve visual acuity, expand field of vision, or slow disease progression for the subject with the retinal disease.6.1 COMPOUNDS

[0070] In some embodiments, the methods of treating provided herein include administering Compound 1 , or a pharmaceutically acceptable form thereof, to a subject. Compound 1 or a pharmaceutically acceptable form thereof is a farnesyltransferase inhibitor, and is a selective farnesyltransferase inhibitor that selectively inhibits farnesyltransferase with greater potency(lower ICso value) relative to the level of inhibition of geranyl geranyl transferase type-1 .

[0071] In some embodiments is Compound 1 (sometimes referred to as the Compound of Formula (I)), which can be named (5)-3-amino-3-(l-methyl-l / / -imidazol-5-yl)-6-oxa-2(4,6)- quinolina-l,4(l,3)-dibenzenacyclohexaphane-22,44-dicarbonitrile, and which has the structure:Compound 1

[0072] The synthesis and certain uses, inhibition activities, and metabolic stabilities of Compound 1 and pharmaceutically acceptable forms thereof, as provided herein, are described in International Patent Publication No. WO 2023 / 102378 Al, the entirety of which is incorporated herein by reference. In some embodiments, the compound for use in the methods of treating provided herein is Compound 1, or a pharmaceutically acceptable form thereof.

[0073] In certain embodiments, the Compound 1 or a pharmaceutically acceptable form thereof is a pharmaceutically acceptable salt of the Compound 1 or a pharmaceutically acceptable solvate thereof. In certain embodiments, the Compound 1 or a pharmaceutically acceptable form thereof is a non-solvate of the Compound 1 or a pharmaceutically acceptable salt thereof. In some embodiments, the Compound 1 or a pharmaceutically acceptable form thereof is an anhydrate of the Compound 1. In some embodiments, the Compound 1 or a pharmaceutically acceptable form thereof is a pharmaceutically acceptable solvate of the Compound 1 or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutically acceptable solvate is selected from the group consisting of: hydrate, a hemihydrate, an iso-butyl acetate solvate, an iso-propyl acetate solvate, a tetrahydrofuran solvate, an acetone solvate, an acetonitrile solvate, or combinations thereof. In some embodiments, the Compound 1 or a pharmaceutically acceptable form thereof is a free base, hemi-hydrate of the Compound 1.

[0074] In some embodiments, provided herein is Form 1 of Compound 1. In some embodiments, Form 1 of Compound 1 is a crystalline free base, hemi-hydrate of Compound 1. In some embodiments, Form 1 of Compound 1 is substantially free of amorphous Compound 1.In some embodiments, Form 1 of Compound 1 is substantially free of other crystalline forms (i.e., polymorphs) of Compound 1. In some embodiments, Form 1 of Compound 1 is substantially free of salt forms of Compound 1. In some embodiments, Form 1 of Compound 1 is provided as substantially pure Form 1 of Compound 1.

[0075] In some embodiments, Form 1 has an enantiomeric purity of about 98%, about 98.5%, about 99%, or about 99.5%, or greater. In some embodiments, Form 1 is crystalline. In some embodiments, Form 1 is substantially crystalline. In some embodiments, Form 1 is about 90%, about 95%, about 96%, about 97%, about 98%, about 98.5%, about 99%, or about 99.5%, crystalline, or greater.

[0076] In some embodiments, Form 1 has an X-ray powder diffraction (XRPD) pattern comprising peaks at approximately 9.0, 12.8, 16.6, and 18.4° 29. In some embodiments, the XRPD pattern further comprises peaks at approximately 8.6, 12.0, 18.1, and 23.2° 20. In some embodiments, the XRPD pattern further comprises peaks at approximately 16.1, 17.1, 24.1, and 25.6° 20. In some embodiments, the XRPD pattern comprises peaks at approximately 8.6, 9.0, 12.0, 12.8, 16.1, 16.6, 17.1, 18.1, 18.4, 23.2, 24.1, and 25.6° 20.

[0077] In some embodiments, provided herein is a solid form comprising a free base, hemihydrate of Compound 1, characterized by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or all of the XRPD peaks located at approximately the following positions (e.g., degrees 20 ± 0.2) when measured using Cu Ka radiation: 8.6, 9.0, 12.0, 12.8, 16.1, 16.6, 17.1, 18.1, 18.4, 23.2, 24.1, and 25.6° 29. In some embodiments, the solid form is characterized by at least 3 of the peaks. In some embodiments, the solid form is characterized by at least 5 of the peaks. In some embodiments, the solid form is characterized by at least 7 of the peaks. In some embodiments, the solid form is characterized by at least 9 of the peaks. In some embodiments, the solid form is characterized by at least 11 of the peaks. In some embodiments, the solid form is characterized by all of the peaks. Form 1 of Compound 1 is described in International Patent Application No.PCT / CN2024 / 096480, the entirety of which is incorporated herein by reference.

[0078] In another aspect is a pharmaceutically acceptable salt of Compound 1, or an isotopologue thereof, or a pharmaceutically acceptable solvate of the pharmaceutically acceptable salt.

[0079] In some embodiments, the pharmaceutically acceptable salt of Compound 1 is a benzoate salt, a besylate salt, a chloride salt, a citrate salt, a fumarate salt, a gentisate salt, aglutarate salt, a glycolate salt, a hippurate salt, a 1 -hydroxy -2-naphthoate salt, a malate salt, a maleate salt, a mesylate salt, an oxalate salt, a phosphate salt, a sulfate salt, a tartrate salt, or a tosylate salt. The pharmaceutically acceptable salts of Compound 1 are described in International Patent Application No. PCT / CN2024 / 096480, the entirety of which is incorporated herein by reference.

[0080] In certain embodiments, the use of the farnesyltransferase inhibitor, in particular Compound 1, and the pharmaceutically acceptable form thereof, is applicable to the farnesyltransferase inhibitor tipifarnib.

[0081] In some embodiments, the methods of treating provided herein include administering (a) Compound 1 or a pharmaceutically acceptable form thereof and (b) a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist) to a subject.

[0082] In some embodiments, the VEGF / VEGFR signaling pathway inhibitor is a VEGFA antagonist. In some embodiments, the VEGFA antagonist is a biologic or monoclonal antibody, such as pegaptanib (Macugen), bevacizumab, ranibizumab, ramucirumab, aflibercept, conbercept, brolucizumab, or faricimab.

[0083] In some embodiments, the VEGFR inhibitor used as provided herein is cabozantinib, lenvantinib, axitinib, regorafenib, vandetanib, pazopanib, sunitinib, sorafenib, tivozanib, fruquintinib, vorolanib, and zanzalintinib, and pharmaceutically acceptable forms thereof. In some embodiments, the VEGFR inhibitor used as provided herein is cabozantinib (S)-malate, lenvantinib mesylate, axitinib free base, regorafenib monohydrate, vandetanib free base, pazopanib hydrochloride, sunitinib (S)-malate, sorafenib tosylate, tivozanib hydrochloride hydrate, fruquintinib free base, or zanzalintinib fumarate.6.2 PHARMACEUTICAL COMPOSITIONS, KITS, AND PACKAGING

[0084] In some embodiments, provided herein is a pharmaceutical composition comprising Compound 1, or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient. In some embodiments, provided herein is a pharmaceutical composition comprising a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist). In some embodiments, the VEGF / VEGFR signaling pathway inhibitor is a VEGFR inhibitor as described herein. In some embodiments, the VEGF / VEGFR signaling pathway inhibitor is a VEGFA antagonist as described herein.

[0085] In some embodiments, provided herein is a pharmaceutical kit comprising Compound1, or a pharmaceutically acceptable form thereof. In some embodiments, provided herein is a pharmaceutical kit comprising: (a) Compound 1, or a pharmaceutically acceptable form thereof; and (b) VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist). In some embodiments, the pharmaceutical kit further comprises instructions that detail a dosing regimen for administering each compound. In some embodiments, the pharmaceutical kit is a pharmaceutical packaging.

[0086] The compounds and pharmaceutical compositions are intended to be administered by a suitable route, including but not limited to orally, parenterally, rectally, topically and locally. The compounds and pharmaceutical compositions provided herein can also be administered intradermally, intramuscularly, intraperitoneally, percutaneously, intravenously, subcutaneously, intravitreally, intranasally, epidurally, sublingually, intracerebrally, transdermally, mucosally, by drops, by inhalation, or topically to the ears, nose, eyes, or skin.

[0087] In some embodiments, the pharmaceutical compositions are provided for administration to a subject in unit dosage forms, such as tablets, capsules, microcapsules, pills, powders, granules, troches, suppositories, injections, port delivery systems, syrups, patches, creams, lotions, ointments, gels, sprays, sterile parenteral solutions or suspensions, and oral solutions or suspensions, and oil water emulsions containing suitable quantities of the therapeutic agents described herein. The pharmaceutical compositions are in liquid, semi-liquid or solid form and are formulated in a manner suitable for each route of administration. For oral administration, capsules and tablets can be formulated. In some embodiments, the pharmaceutical compositions provided herein are in the form of a tablet. In some embodiments, the pharmaceutical compositions provided herein are in the form of a capsule. In some embodiments, administration is via a port delivery system or a hydrogel, and may be applied directly to the eye.

[0088] Typically, the compound disclosed herein is formulated into pharmaceutical compositions using techniques and procedures well known in the art (see, e.g., Ansel Introduction to Pharmaceutical Dosage Forms, Seventh Edition 1999). In some embodiments, the pharmaceutical compositions are formulated and administered in unit dosage forms or multiple dosage forms. Such dosage forms contain predetermined amounts of active ingredients, and may be prepared by methods of pharmacy well known to those skilled in the art. Unit dose forms as used herein refer to physically discrete units suitable for human and animal subjects andpackaged individually as is known in the art. Each unit dose contains a predetermined quantity of the therapeutically active compound sufficient to produce the desired therapeutic effect, in association with the required pharmaceutical excipient. Examples of unit dose forms include ampules and syringes and individually packaged tablets or capsules. Unit dose forms may be administered in fractions or multiples thereof. A multiple dose form is a plurality of identical unit dosage forms packaged in a single container to be administered in segregated unit dose form. Examples of multiple dose forms include vials, bottles of tablets or capsules or bottles of pints or gallons. Hence, multiple dose form is a multiple of unit doses which are not segregated in packaging.

[0089] The compounds and pharmaceutical compositions provided herein may be administered at once, or may be divided into a number of smaller doses, to be administered at intervals of time. It is understood that the precise dosage and duration of treatment is a function of the disease (e.g., a retinal disease) being treated. It is to be noted that concentrations and dosage values may also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens may be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the pharmaceutical compositions, and that the concentration ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed pharmaceutical compositions.

[0090] The mode of administration is left to the discretion of the healthcare practitioner, and can depend in-part upon the site of the medical condition. Compound 1, or a pharmaceutically acceptable form thereof, and / or the VEGF / VEGFR signaling pathway inhibitor (e.g., the VEGFR inhibitor or VEGFA antagonist), may be formulated, alone or together, in suitable dosage unit with pharmaceutically acceptable excipients, such as carriers, adjuvants, and vehicles, as appropriate for each route of administration.6.3 METHODS, DOSING REGIMENS AND SCHEDULES6.3.1 THERAPEUTIC METHODS

[0091] In some embodiments, provided herein is a method of treating a retinal disease in a subject comprising administering to the subject Compound 1, or a pharmaceutically acceptable form thereof (or a pharmaceutical composition comprising the same). In some embodiments, provided herein is a method of treating a retinal disease in a subject comprising administering tothe subject Compound 1, or a pharmaceutically acceptable form thereof (or a pharmaceutical composition comprising the same), and a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist). In some embodiments, provided herein is a method of treating a retinal disease in a subject comprising administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable form thereof (or a pharmaceutical composition comprising the same). In some embodiments, provided herein is a method of treating a retinal disease in a subject comprising administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable form thereof (or a pharmaceutical composition comprising the same), and an effective amount of a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist).

[0092] In some embodiments, the subject to whom the compounds are administered in the methods provided herein has, suffers from, has symptoms associated with, or is diagnosed as having, a retinal disease. In some embodiments, the subject has or suffers from a retinal disease. In some embodiments, the subject has symptoms associated with a retinal disease. In some embodiments, the subject is diagnosed as having a retinal disease. In some embodiments, the subject is a previously treated retinal disease subject. In some embodiments, the subject may be diagnosed as having a retinal disease by one skilled in the art, for example, a physician, such as an ophthalmologist.

[0093] In some embodiments, the method comprises administering the Compound 1, or the pharmaceutically acceptable form thereof, to the subject orally.

[0094] In some embodiments, the method comprises administering the Compound 1, or the pharmaceutically acceptable form thereof, to the subject intravenously.

[0095] In some embodiments, the method comprises administering the Compound 1, or the pharmaceutically acceptable form thereof, to the subject intravitreally.

[0096] In some embodiments, the method comprises administering the VEGF / VEGFR signaling pathway inhibitor (e.g., the VEGFR inhibitor or the VEGFA antagonist) to the subject orally, optionally once or twice daily, optionally for one or more treatment cycles.

[0097] In some embodiments, the method comprises administering the VEGF / VEGFR signaling pathway inhibitor (e.g., the VEGFR inhibitor or the VEGFA antagonist) to the subject intravenously, optionally once or twice daily, optionally for one or more treatment cycles.

[0098] In some embodiments, the method comprises administering the VEGF / VEGFRsignaling pathway inhibitor (e g., the VEGFR inhibitor or the VEGFA antagonist) to the subject intravitreally, optionally once or twice daily, optionally for one or more treatment cycles.

[0099] In some embodiments, the retinal disease is selected from a group consisting of diabetic retinopathy, diabetic macular edema (DME), age-related macular degeneration (AMD), retinal vein occlusion (RVO), and retinopathy of prematurity. In some embodiments, the retinal disease is diabetic retinopathy. In some embodiments, the retinal disease is diabetic macular edema (DME). In some embodiments, the retinal disease is diabetic retinopathy. In some embodiments, the retinal disease is age-related macular degeneration (AMD). In some embodiments, the retinal disease is retinal vein occlusion (RVO). In some embodiments, the retinal disease is retinopathy of prematurity. In some embodiments, the retinal disease is diagnosed according to the International Classification of Diseases, Tenth Revision (ICD-10), codes H00-H59, or related sections in subsequent revisions. In some embodiments, the retinal disease is diagnosed following one or more ocular tests selected from dilated eye examination, fluorescein angiography, optical coherence tomography, visual acuity test, tonometry, ophthalmoscopy, Amsler grid test, optical coherence tomography angiography, and el ectroretinography .

[0100] In some embodiments, treatment with Compound 1, or a pharmaceutically acceptable form thereof, affects protein expression of one or more endothelial cell protein markers, which function in endothelial cell migration, growth, stability, permeability, and / or maturation. In some embodiments, treatment with Compound 1, or a pharmaceutically acceptable form thereof, reduces expression of one or more of the following endothelial cell protein markers: Table 1.

[0101] In some embodiments, treatment with Compound 1, or a pharmaceutically acceptable form thereof, reduces expression of CD31 in endothelial cells. In some embodiments, treatment with Compound 1, or a pharmaceutically acceptable form thereof, reduces expression of VEGFR2 in endothelial cells. In some embodiments, treatment with Compound 1, or a pharmaceutically acceptable form thereof, increases expression of IGFBP3in endothelial cells.

[0102] In some embodiments, Compound 1, or a pharmaceutically acceptable form thereof, that is administered according to the methods provided herein inhibits famesylation of a protein, for example inhibits famesylation of a farnesylation-dependent protein. Without being bound by any one theory, in some embodiments, Compound 1, or a pharmaceutically acceptable form thereof, administered according to the methods provided herein inhibits famesylation of one or more farnesylation-dependent proteins selected from RhoB, RhoE, and Lamin B, or a combination thereof. In some embodiments, the farnesylation-dependent protein is a dysregulated farnesylation-dependent protein.

[0103] In some embodiments, inhibition of the famesylation of the farnesylation-dependent protein, according to the methods of treating provided herein, occurs in a cell, such as in a cell of the subject. In some embodiments, the cell is a mammalian cell. In some embodiments, the cell a human cell.

[0104] In some embodiments, administering Compound 1, or a pharmaceutically acceptable form thereof, in combination with a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist), provides a therapeutic benefit, such as a synergistic benefit, to the subject: (a) relative to, for combinations with a VEGF / VEGFR signaling pathway inhibitor (e g., a VEGFR inhibitor or a VEGFA antagonist), or VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist) monotherapy, such as relative to treatment of a retinal disease with a VEGFR inhibitor or VEGFA antagonist therapy in the retinal disease subject; (b) relative to standard of care treatment for the retinal disease, including but not limited to surgery, radiofrequency ablation, radiation therapy, laser treatment, vitrectomy, non-steroidalanti -inflammatory drugs, or chemotherapy, or combinations thereof; or (c) relative to no treatment for the retinal disease.

[0105] In some embodiments, the methods comprise administering to the subject a pharmaceutical composition comprising Compound 1, or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient. In some embodiments, the methods comprise administering to the subject: (a) a pharmaceutical composition comprising Compound 1, or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient; and (b) a pharmaceutical composition comprising a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist) and a pharmaceutically acceptable excipient. For example, in some embodiments, the methods comprise administering to the subject a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient. In some embodiments, the methods comprise administering to the subject: (a) a pharmaceutical composition comprising an effective amount of Compound 1, or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient; and (b) a pharmaceutical composition comprising an effective amount of a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor or a VEGFA antagonist) and a pharmaceutically acceptable excipient.

[0106] In some embodiments, the methods provided herein comprise administering to the subject a pharmaceutical kit or pharmaceutical packaging comprising a pharmaceutical composition comprising Compound 1, or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient.6.3.2 DOSES AND REGIMENS

[0107] In some embodiments, the methods provided herein comprise administering to the subject Compound 1, or a pharmaceutically acceptable form thereof. For example, in some embodiments, the methods provided herein comprise administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable form thereof. In some embodiments, the methods provided herein comprise administering to the subject: (a) Compound 1, or a pharmaceutically acceptable form thereof; and (b) a VEGF / VEGFR signaling pathway inhibitor (e.g., a VEGFR inhibitor, such as cabozantinib, lenvantinib, axitinib, regorafenib, vandetanib, pazopanib, sunitinib, sorafenib, tivozanib, fruquintinib, vorolanib, or zanzalintinib, such as cabozantinib, or a pharmaceutically acceptable form thereof; or a VEGFA antagonist, such as abiologic or monoclonal antibody, such as pegaptanib (Macugen), bevacizumab, ranibizumab, ramucirumab, aflibercept, conbercept, brolucizumab, or faricimab). In some embodiments, the methods provided herein comprise administering to the subject (a) an effective amount of Compound 1, or a pharmaceutically acceptable form thereof, and (b) an effective amount of the VEGF / VEGFR signaling pathway inhibitor, or a pharmaceutically acceptable form thereof. In some embodiments, the methods comprise administering to the subject pharmaceutical compositions of each agent as described herein. In some embodiments, the methods comprise administering to the subject pharmaceutical compositions comprising an effective amount of each agent as described herein.

[0108] In some embodiments, Compound 1, or a pharmaceutically acceptable form thereof, is administered to the subject according to the methods provided herein at a dose of 0.5 to 20 mg per day, or 0.5 to 10 mg per day. In some embodiments, the per day dose is 0.005 mg / 0.05 mL vehicle to 4 mg / 0.10 mL vehicle per day for intravitreal injection. In some embodiments, Compound 1, or a pharmaceutically acceptable form thereof, is administered 1, 2, 3, or 4 times per day. In some embodiments, the per day dose of Compound 1, or a pharmaceutically acceptable form thereof, is split into two, three, or four doses, such as two, three, or four equal doses, and particularly two doses or two equal doses, that are administered to the subject according to the methods provided herein. In some embodiments, Compound 1, or a pharmaceutically acceptable form thereof, is administered once or twice per day, or is administered once per day, or is administered twice per day. In some embodiments, Compound 1 or a pharmaceutically acceptable form thereof is administered via a port delivery system, optionally administered continuously via the port delivery system. In some embodiments, the dosing is for one or more treatment cycles, such as 28-day treatment cycles. In some embodiments, Compound 1, or a pharmaceutically acceptable form thereof, is administered intermittently, such as 3 to 14 days on / 3 to 14 days off, or 7 days on / 7 days off, or 3 weeks on / 1 week off, or 1 week on / 3 weeks off, optionally in 28-day treatment cycles (such as on days 1-7 and 15-21, or days 8-14 and 22-28, or days 1-21, of a 28-day treatment cycle).

[0109] In some embodiments, the VEGFR inhibitor, cabozantinib, lenvantinib, axitinib, regorafenib, vandetanib, pazopanib, sunitinib, sorafenib, tivozanib, fruquintinib, vorolanib, or zanzalintinib, or a pharmaceutically acceptable form thereof, is administered to the subject according to the methods of treating provided herein at a dose of 0.2 to 1500 mg per day. Insome embodiments, the dose of the VEGFR inhibitor, cabozantinib, lenvantinib, axitinib, regorafenib, vandetanib, pazopanib, sunitinib, sorafenib, tivozanib, fruquintinib, vorolanib, or zanzalintinib, or a pharmaceutically acceptable form thereof, administered to the subject is selected from 0.5-10 mg, 2-15 mg, 10-30 mg, 10-40 mg, 10-240 mg, 20-50 mg, 20-240 mg, 30- 50 mg, 35-70 mg, 40-80 mg, 60-100 mg, 80-120 mg, 80-160 mg, 80-240 mg, 160-250 mg, 160- 300 mg, 100-600 mg, or 200-1000 mg mg, per day. In some embodiments, the dose of the VEGFR inhibitor, cabozantinib, lenvantinib, axitinib, regorafenib, vandetanib, pazopanib, sunitinib, sorafenib, tivozanib, fruquintinib, or zanzalintinib, administered to the subject is selected from (Table 2):Table 2

[0110] In some embodiments, the VEGFR inhibitor, cabozantinib, lenvantinib, axitinib, regorafenib, vandetanib, pazopanib, sunitinib, sorafenib, tivozanib, fruquintinib, vorolanib, or zanzalintinib, or a pharmaceutically acceptable form thereof, is administered 1, 2, 3, or 4 times per day. In some embodiments, the per day dose of the VEGFR inhibitor is split into two amounts, such as two equal amounts, that are administered to the subject according to the methods provided herein. In some embodiments, the VEGFR inhibitor is administered once or twice per day, such as once per day. In some embodiments, the dose of the VEGFR inhibitor is administered to the subject daily for one or more treatment cycles according to the methods provided herein.

[0111] In some embodiments, the VEGFA antagonist, pegaptanib (Macugen), bevacizumab, ranibizumab, ramucirumab, aflibercept, conbercept, brolucizumab, or faricimab is administered intravitreally. Pegaptanib is 0.3 mg (via 0.3 mg / 90 pL) once every six weeks. For example, bevacizumab (Avastin) may be administered at 5 mg / kg to 15 mg / kg via injection of a 25 mg / mL solution; ranibizumab (Susvimo; or ranibizumab-nuna) may be administered intravitreally via an ocular implant, at 2 mg (100 mg / mL solution) delivered continuously via the implant over 24 weeks, optionally with additional 0.5 mg intravitreal injection as needed; ramucirumab (Cyramza) may be administered by intravenous infusion of 8 to 10 mg / kg every 2 or 3 weeks via a 10 mg / mL solution; aflibercept (Eylea) may be administered by intravitreal injection of 0.4 to 2 mg (via a 40 mg / mL solution) every 10 days, or every 4, 8, or 12 weeks; conbercept may be administered at 0.5 mg to 2 mg intravitreally once a month, or as needed; brolucizumab (Beovu; or brolucizumab-dbll) may be administered at 6 mg every 4 to 12 weeks by intravitreal injection; faricimab (Vabysmo; or faricimab-svoa) may be administered by intravitreal injection at 6 mg (via a 120 mg / mL solution) every 4 weeks / monthly, or every 8, 12, or 16 weeks.

[0112] In some embodiments, Compound 1, or a pharmaceutically acceptable form thereof, and the VEGF / VEGFR signaling pathway inhibitor (e.g., the VEGFR inhibitor or VEGFA antagonist), are administered to the subject on different dosing schedules. For example, Compound 1, or a pharmaceutically acceptable form thereof, may be administered on an intermittent dosing schedule while the VEGF / VEGFR signaling pathway inhibitor isadministered on a continuous dosing schedule, such as 1, 2, 3, or 4 times daily, or the two agents are administered through different routes and schedules, such as one of the two agents administered by infusion and the other agent administered orally.7. EXAMPLES

[0113] The following Examples are presented by way of illustration, not limitation.

[0114] EXAMPLE 1: Combination Study in Cell Line-Derived Xenograft (CDX) and Patient-Derived Xenograft (PDX) in vivo Models

[0115] Clear cell renal cell carcinoma (ccRCC) tumors frequently have a genetic deletion of VHL, which leads to the stabilization of HIFloc / 2oc proteins, key transcription factors that drive tumor angiogenesis. As such, ccRCC tumors are highly vascularized and are currently treated with antiangiogenic TKIs. Compound 1 was tested in ccRCC preclinical models. These results, combined with evidence of anti-angiogenic activity for Compound 1 as provided below, indicate that Compound 1, or a pharmaceutically acceptable form thereof, is useful in the treatment of diseases dependent on angiogenesis, such as retinal diseases.

[0116] VHL-mutant 786-0 tumor cells were maintained in vitro in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS). VHL-mutant A498 tumor cells were maintained in vitro in Eagle’s Minimum Essential Medium supplemented with 10% FBS. Cells were grown at 37 °C in an atmosphere of 5% CO2 in air. Cells were harvested while in exponential growth phase and quantified by cell counter before tumor inoculation. For the VHL mutant KI-0326 and KI-12-0073 ccRCC PDX models, fresh tumor tissues from mice bearing established primary human cancer tissues were harvested and cut into small pieces (approximately 2-3 mm in diameter). Each female BALB / c mouse was inoculated subcutaneously in the right upper flank region with the tumor cells at 5 x 106per mouse (CDX models) in 0.1 mL of phosphate-buffered saline (PBS) or was inoculated surgically (approx. 30 mm3slice; PDX models) for tumor development. Randomization started when mean tumor size reached approximately 250-300 mm3. Animals were randomly allocated to study groups at 5-6 animals per study group, depending on the study design, based on “Matched distribution” method / “ Stratified” method (Study Director™ software, version 3.1.399.19) / randomized block design. Administration of test articles was initiated on the same day as randomization.Compound 1 was administered at 20 mg / kg twice daily, cabozantinib was administered at 8, 15or 20 mg / kg p.o. once daily, and axitinib was administered at 36 mg / kg p.o., once daily. Animals were checked daily for morbidity and mortality after tumor cell inoculation. During routine monitoring, the animals were checked for any effects on tumor growth, on behavior, including mobility, food and water consumption, and on physical characteristics, including body weight gain / loss, eye / hair matting and any other abnormalities. Mortality and observed clinical signs were recorded for individual animal. Body weights and tumor volumes were measured twice per week after randomization. For tumor volumes, measurements were performed in two dimensions using a caliper and recorded in mm3using the formula: V = (L x W x W) / 2, where V is tumor volume, L is tumor length (the longest tumor dimension) and W is tumor width (the longest tumor dimension perpendicular to L). Dosing as well as tumor and body weight measurements were conducted in a Laminar Flow Cabinet. Body weights and tumor volumes were measured using StudyDirector™ software, version 3.1.399.19. At study termination, tumors were harvested by taking a section for formalin fixing and paraffin embedding (FFPE) and snap freezing the rest. FFPE was performed using standard procedures.

[0117] As shown in FIG. 1, A498 CDX continuously treated with Compound 1 (20 mg / kg, BID) and axitinib (36 mg / kg, QD) showed an increase in tumor regression relative to either agent alone. Error bars represent the standard error of the mean (n = 5).

[0118] As shown in FIG. 2 for KL 12-0073 PDX, continuous treatment with Compound 1 (20 mg / kg, BID) and axitinib (36 mg / kg, QD) resulted in increased tumor growth inhibition compared to either agent alone. Error bars represent the standard error of the mean (n = 8).

[0119] As shown in FIG. 3, 786-0 CDX (FIG. 3A) continuously treated with Compound 1 (20 mg / kg, BID) and cabozantinib (20 mg / kg, QD) (lane 3) showed increased percent tumor growth inhibition (% TGI) relative to Compound 1 (lane 1) or cabozantinib (lane 2) alone. A498 CDX (FIG. 3B) continuously treated with Compound 1 (20 mg / kg, BID) and cabozantinib (8 or 20 mg / kg, QD) (lanes 4 and 5, respectively) showed increased % TGI compared to Compound 1 alone (20 mg / kg, BID) (lane 1) or cabozantinib alone (8 and 20 mg / kg, QD) (lanes 2 and 3, respectively). Error bars represent the standard error of the mean (786-0, n = 6, % TGI calculated 16 days post start of treatment; A498, n = 5, % TGI calculated 14 days post-start of treatment). % TGI was calculated with the following formula: [l-(mean volume of treated tumors) / (mean volume of control tumors)] x 100%.

[0120] As shown in FIG. 4, mice with KI- 12-0073 VHL-mutant PDX, 786-0 VHL-mutantCDX, and KI-0326 VHL-mutant PDX treated continuously with Compound 1 (20 mg / kg, BID) and cabozantinib (8, 15, and 20 mg / kg, QD, respectively) exhibited reduced tumor growth (FIGS. 4A, 4C, and 4E), compared to either compound alone. Error bars represent the standard error of the mean. FIGS. 4B, 4D, and 4F show the graphs of percent of tumor volume change at endpoint relative to day 0 in the KI- 12-0073,786-0, and KI-0326 models, respectively, to show the variability in responses to cabozantinib, compared to the combination of compound of Formula (I) and cabozantinib, which resulted in regression of all but one tumor.

[0121] As shown in FIG. 5, mice with 786-0 VHL-mutant CDX continuously treated with Compound 1 (20 mg / kg, BID) and varying doses of cabozantinib (4, 8, 10, and 12 mg / kg, QD) showed dose-dependent tumor growth inhibition compared to the respective single agent cabozantinib or Compound 1. Percent tumor volume change was calculated using endpoint tumor volume values (day 28) relative to day 0.

[0122] Mice with 786-0 CDX were treated continuously with cabozantinib (15 mg / kg, QD), Compound 1 (20 mg / kg, BID), lenvatinib (10 mg / kg, QD), lenvatinib plus everolimus (2 mg / kg, QD), Compound 1 plus cabozantinib, and Compound 1 with lenvatinib. As shown in FIG. 6, the combination of Compound 1 (20 mg / kg, BID) with either cabozantinib or lenvatinib reduced tumor growth more than any of the agents alone, and exhibited reduced tumor growth that compared favorably to treatment with lenvatinib (10 mg / kg, QD) and everolimus (kinase / mTOR inhibitor; 2 mg / kg, QD), which combination is an FDA-approved second-line treatment for RCC. Error bars represent the standard error of the mean.

[0123] Mice with 786-0 CDX were treated continuously with axitinib (36 mg / kg, QD) for 14 days. Starting at day 15, animals were dosed with one of the following: a) vehicle; b) Compound 1 (20 mg / kg, BID); (c) cabozantinib (15 mg / kg, QD); (d) axitinib (36 mg / kg, QD); or (e) the combination of Compound 1 and cabozantinib. As shown in FIG. 7, 786-0 CDX that progressed during the 14-day treatment with axitinib and then were treated continuously with Compound 1 plus cabozantinib exhibited reduced tumor growth compared to any of the other arms over the subsequent 18 days. Results for dosing for a total of three to four weeks following axitinib pre-treatment may provide similar results. Error bars represent the standard error of the mean.

[0124] EXAMPLE 2: Mechanism Studies

[0125] Study 1. To investigate the mechanism of action of the results of Example 1, VHL-mutant RCC cell lines can be subjected to hypoxia (1% O2) in vitro to mimic the hypoxic conditions induced by anti-angiogenic TKIs in vivo and may be treated with Compound 1 to evaluate its impact on signaling pathways in hypoxia-exposed cells. In this study, hypoxia may initially reduce mTOR signaling, but it may rebound after 24 hours in hypoxia, in which case it indicates that mTOR pathway reactivation is a potential mechanism of resistance to TKIs. Addition of Compound 1, or a pharmaceutically acceptable form thereof, may block hypoxia- induced mTOR reactivation. Mechanistically, Compound 1 potently inhibits the famesylation, and hence the activity, of an obligate famesylated protein RHEB, a positive regulator of mTOR, suggesting that the synergy may arise through RHEB inhibition in this model. The mechanistic data in cell lines suggest that the ability of Compound 1, or a pharmaceutically acceptable form thereof, to inhibit mTOR reactivation observed in ccRCC cell lines under hypoxic stress may contribute to enhanced treatment durability in vivo.

[0126] Study 2. 786-0 CDX were snap frozen after 14 days of treatment with vehicle, cabozantinib (15 mg / kg, QD), Compound 1 (20 mg / kg, BID), or the combination. Tumors were thawed in IX RIPA buffer (Thermo Scientific Cat # PI89901) supplemented with IX HALT protease and phosphatase inhibitor cocktail (Thermo Scientific Cat # PI78446) then homogenized using a bead mill homogenizer for 30 seconds at 4.5 m / s. Lysates were clarified by centrifugation for 10 min at 12k x g and quantified by BCA assay (Pierce). For SDS-PAGE and immunoblotting, 20-50 pg of lysate was loaded on to 4-12% Bis-Tris gels (Invitrogen NuPAGE) and transferred on to nitrocellulose membranes. Membranes were probed with the following antibodies: anti-phospho-ERKl / 2 (CST Cat # 4695); anti-phospho-AKT (CST Cat # 4060); anti-phospho-S6 (Ser235 / 236) (CST Cat # 2211); anti-phospho-S6 (Ser240 / 244) (CST Cat # 2215); anti-total S6 (CST Cat # 2217); anti-phospho-RB (CST Cat # 8516); anti-cyclin DI (CST Cat # 55506); anti-RHEB (CST Cat # 13879) and anti-HSP90 (CST Cat # 4877). As shown in FIG. 8, treatment with the combination led to decreased phosphorylation of AKT and S6, two growth-promoting signaling proteins, and decreased phosphorylated RB, a cell cycle arrest marker, compared to either agent alone. A slight shift of RHEB was detected in the combination treatment, indicating defarnesylation by Compound 1. HSP90 served as loading control.

[0127] Study 3. All immunohistochemistry (IHC) stains were performed at Histowiz, Inc. (Brooklyn, NY) using the Leica BOND RX automated Stainer (Leica Microsystems). The slideswere dewaxed using xylene- and alcohol-based dewaxing solutions. Epitope retrieval was performed by heat-induced epitope retrieval (HIER) of the formalin-fixed, paraffin-embedded tissue in citrate-based pH 6 solution for 20 min at 95 °C. The tissues were first incubated with peroxide block buffer (Leica Microsystems), followed by incubation with the primary antibody at 1 : 100 dilution for 30 min, followed by DAB mouse secondary reagents: polymer, DAB refine, and hematoxylin (Leica Microsystems). The slides were dried, cover-slipped and visualized using a Leica Aperio AT2 slide scanner (Leica Microsystems). The following primary antibodies were used: anti-CD31 antibody (Sigma, 131M-94) and anti-VEGFR2 antibody (Cell Signaling Technology 9698).

[0128] As shown in Table 3, consistent with the anti angiogenic activity of cabozantinib, 786- O CDX treated with cabozantinib (15 mg / kg, QD) for 14 days resulted in decreased angiogenesis, which is evident as decreased expression of CD31 and VEGFR2, compared to vehicle. The combination of Compound 1 (20 mg / kg, BID) and cabozantinib (15 mg / kg, QD), however, led to greater reductions of CD31 and VEGFR2 expression, compared to cabozantinib alone.Table 3.

[0129] As shown in Table 4, KI-0326 PDX treated with cabozantinib (20 mg / kg, QD) for 14 days led to reduced tumor vascularity compared to vehicle, as measured by CD31 immunohistochemistry. However, the combination of cabozantinib and Compound 1 (20 mg / kg, BID) did not lead to greater reduction of CD31 expression. This suggests that the additive effectof the two drugs on tumor growth inhibition is not solely driven by the inhibition of angiogenesis.Table 4.

[0130] Study 4. Early passage (less than passage 6) human umbilical vein endothelial cells (HUVEC) were seeded at 1,000 cells per well on a 96-well plate in endothelial cell medium with 0.2% fetal bovine serum (FBS) and allowed to sit overnight. The next day, media was replaced with fresh endothelial cell medium with 5% fetal bovine serum (FBS), 100 ng / mL recombinant VEGFA, and endothelial cell growth supplement (ECGS) plus test article: DMSO as vehicle, varying concentrations of axitinib with or without 100 nM Compound 1; or varying concentrations of cabozantinib with or without 100 nM Compound 1. At Day 5, cell viability was assayed using Cell Titer-Gio 2 reagent (Promega) per manufacturer’s instructions, with luminescence recorded on the Tecan plate reader.

[0131] As shown in Table 5, addition of 100 nM Compound 1 to axitinib or cabozantinib resulted in more potent inhibition of HUVEC proliferation, with reduced IC50 concentrations compared to axitinib or cabozantinib alone.Table 5.

[0132] Study 5. Early passage (less than passage 6) human umbilical vein endothelial cells (HUVEC) were seeded at 1,000 cells per well on a 96-well plate in complete endothelial cell medium containing 5% fetal bovine serum (FBS) and endothelial cell growth supplement (ECGS) and allowed to sit overnight. The next day, test articles were added: DMSO as vehicle, varying concentrations of cabozantinib, axitinib, or lenvatinib, and varying concentrations of Compound 1. At Day 7 post-test article addition, cell viability was assayed using Cell Titer-Gio 2 reagent (Promega) per manufacturer’s instructions, with luminescence recorded on the Tecan plate reader.

[0133] As shown in FIG. 9, addition of increasing doses of Compound 1 to cabozantinib (FIG. 9A), axitinib (FIG. 9B), or lenvatinib (FIG. 9C) resulted in more potent inhibition of HUVEC viability compared to the respective TKI agent alone. Additionally, Compound 1 inhibited HUVEC viability in vitro as a single agent with an ICso of 223.4 ± 84.02 nM.

[0134] Study 6. Early passage (less than passage 6) HUVEC or GFP-labeled HUVEC cells were serum-starved overnight and seeded the following day at 6 x 104cells per well on a 48-well plate that was pre-coated with a layer of reduced growth factor base membrane extract (BME). At plating, cells were treated with DMSO (vehicle), 100 nM axitinib, 10 nM cabozantinib, 300 nM or 1 pM Compound 1, 100 nM axitinib plus 1 pM Compound 1, or 10 nM cabozantinib plus 300 nM or 1 pM Compound 1. Each treatment group had two technical replicates. Plates were incubated in an Incucyte at 37 °C in an atmosphere of 5% CO2 in air. Tube formation was monitored by imaging every 30 min for 18 h.

[0135] As shown in FIG. 10, treatment of primary endothelial cells with 100 nM axitinib or 10 nM cabozantinib with or without 1 pM Compound 1 compromised the cells’ ability to form tubular structures on matrix proteins in vitro. (A - vehicle; B - axitinib; C -cabozantinib; D - Compound 1; E - axitinib and Compound 1; F - cabozantinib and Compound 1.) FIG. 11 shows that treatment with 10 nM cabozantinib inhibited in vitro tube formation of GFP-labeled primary endothelial cells while 300 nM Compound 1 did not (FIG. HA, GFP imaging; FIG. 11B, plotsof number of master segments and total length of master segments). The combination of cabozantinib and Compound 1 did not further decrease tube formation compared to cabozantinib alone, demonstrating that Compound 1 does not affect this particular endothelial cell function.

[0136] Study 7. Early passage (less than passage 6) human umbilical vein endothelial cells (HUVEC) were seeded at 2,000 cells per well on a Nunc 96-well, flat-bottom plate in endothelial cell growth factor-supplemented endothelial cell medium (ECM) with 5% fetal bovine serum (FBS) and allowed to sit overnight. The next day, the media was removed and replaced with Incucyte Annexin V Orange dye (Sartorius) diluted (1 :200) in complete ECM media. The following test articles were added directly into Annexin V prepared media: DMSO as vehicle, 1000 nM staurosporine as positive control, 100 nM Compound 1, 100 nM cabozantinib, or combination of Compound 1 and cabozantinib. Live-cell imaging and analysis was done for four days using an Incucyte SX5 system.

[0137] As shown in FIG. 12, treatment of primary endothelial cells with Compound 1 and cabozantinib induced more apoptosis than either agent alone, as measured by Annexin V signal plotted with time. Staurosporine was included as positive control.

[0138] EXAMPLE S: Single Agent In Vivo Studies

[0139] Cakil CDX tumor cells were maintained in vitro in McCoy’s 5A medium supplemented with 10% fetal bovine serum (FBS). Cells were grown at 37 °C in an atmosphere of 5% CO2 in air. Cells were harvested while in exponential growth phase and quantified by cell counter before tumor inoculation. For the KI-12-0097, KI-12-0351, and KI-12-0073 ccRCC PDX models, fresh tumor tissues from mice bearing established primary human cancer tissues were harvested and cut into small pieces (approximately 2-3 mm in diameter). Each female BALB / c mouse was inoculated subcutaneously in the right upper flank region with the tumor cells at 5 x 106per mouse (CDX models) in 0.1 m of phosphate-buffered saline (PBS) or was inoculated surgically (approx. 30 mm3slice; PDX models) for tumor development.Randomization started when mean tumor size reached approximately 250-300 mm3. Animals were randomly allocated to study groups at 6-8 animals per study group, depending on the study design, based on “Matched distribution” method / “ Stratified” method (StudyDirector™ software, version 3.1.399.19) / randomized block design. Administration of test articles was initiated on the same day as randomization. Compound 1 was administered at 20 mg / kg twice daily and cabozantinib was administered at 8 or 10 mg / kg p.o. once daily, and axitinib was administered at36 mg / kg p.o., once daily. Animals were checked daily for morbidity and mortality after tumor cell inoculation. During routine monitoring, the animals were checked for any effects on tumor growth, on behavior, including mobility, food and water consumption, and on physical characteristics, including body weight gain / loss, eye / hair matting and any other abnormalities. Mortality and observed clinical signs were recorded for individual animal. Body weights and tumor volumes were measured twice per week after randomization. For tumor volumes, measurements were performed in two dimensions using a caliper and recorded in mm3using the formula: V = (L x W x W) / 2, where V is tumor volume, L is tumor length (the longest tumor dimension) and W is tumor width (the longest tumor dimension perpendicular to L). Dosing as well as tumor and body weight measurements were conducted in a Laminar Flow Cabinet. Body weights and tumor volumes were measured using StudyDirector™ software, version 3.1.399.19. At study termination, tumors were harvested by taking a section for formalin fixing and paraffin embedding (FFPE) and snap freezing the rest. FFPE was performed using standard procedures.

[0140] As shown in FIG. 13, in each of the above-mentioned models, Compound 1 as a single agent provided comparable activity to cabozantinib.

[0141] EXAMPLE 4: Mechanistic Studies

[0142] To investigate the mechanism of action of Compound 1 in endothelial cells, early passage (less than passage 6) human umbilical vein endothelial cells (HUVEC) was subjected to serum starvation and treatment with Compound 1, cabozantinib, or the combination for two hours. The cells were stimulated with recombinant VEGFA ligand, collected at different timepoints, and assessed for expression of VEGFA-dependent signaling proteins and cell cycle arrest markers using immunoblotting.

[0143] As shown in FIG. 14, Compound 1 negatively impacted mTORCl signaling (reduced phosphorylation of S6 and 4EBP1) and induced cell cycle arrest (reduced phosphorylation of Rb) in VEGFA-stimulated HUVEC cells. The effect on mTORCl may stem from the ability of Compound 1 to inhibit farnesylation of RHEB, a modification necessary for the proper localization and activity of the protein, and of other famesylated targets that play key roles in angiogenesis.INCORPORATION BY REFERENCE

[0144] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference in their entireties to the same extent as if each individualpublication, patent, or patent application was specifically and individually indicated to be incorporated by reference in its entirety. In case of conflict, the present application, including any definitions herein, will control.

Claims

CLAIMSWhat is claimed is:

1. A method of treating a retinal disease in a subject, comprising administering Compound 1 :(Compound 1), or a pharmaceutically acceptable form thereof, to a subject having a retinal disease.

2. A method of inhibiting angiogenesis in a subject, comprising administering Compound 1 :(Compound 1), or a pharmaceutically acceptable form thereof.

3. A method of inhibiting VEGF / VEGFR signaling pathway in a subject, comprising administering Compound 1 :(Compound 1), or a pharmaceutically acceptable form thereof.

4. The method of claim 2 or 3, wherein the subject has a retinal disease.

5. The method of any one of claims 1-4, wherein the method comprises administering a VEGF / VEGFR signaling pathway inhibitor, optionally wherein the VEGF / VEGFR signalingpathway inhibitor is a VEGFR inhibitor or a VEGFA antagonist.

6. The method of claim 5, wherein the VEGF / VEGFR signaling pathway inhibitor is:(a) a VEGFR inhibitor optionally selected from cabozantinib, lenvantinib, axitinib, regorafenib, vandetanib, pazopanib, sunitinib, sorafenib, tivozanib, fruquintinib, vorolanib, and zanzalintinib, and pharmaceutically acceptable forms thereof; optionally wherein the VEGFR inhibitor is selected from cabozantinib (S)-malate, lenvantinib mesylate, axitinib free base, regorafenib monohydrate, vandetanib free base, pazopanib hydrochloride, sunitinib (S)-malate, sorafenib tosylate, tivozanib hydrochloride hydrate, fruquintinib free base, or zanzalintinib fumarate; optionally wherein the VEGFR inhibitor is cabozantinib, axitinib, sunitinib, or sorafenib, or a pharmaceutically acceptable form thereof; optionally wherein the VEGFR inhibitor is cabozantinib or a pharmaceutically acceptable form thereof; optionally wherein the VEGFR inhibitor is zanzalintinib or fruquintinib or a pharmaceutically acceptable form thereof; or(b) a VEGFA antagonist optionally selected from a biologic or monoclonal antibody, optionally selected from pegaptanib, bevacizumab, ranibizumab, ramucirumab, aflibercept, conbercept, brolucizumab, and faricimab.

7. The method of any one of claims 1 or 4-6, wherein the retinal disease is selected from diabetic retinopathy, diabetic macular edema (DME), age-related macular degeneration (AMD), retinal vein occlusion (RVO), or retinopathy of prematurity.

8. The method of any one of claims 1-7, wherein the Compound 1, or a pharmaceutically acceptable form thereof, is administered orally to the subject.

9. The method of any one of claims 1-7, wherein the Compound 1, or a pharmaceutically acceptable form thereof, is administered intravenously to the subject.

10. The method of any one of claims 1-9, wherein the Compound 1, or a pharmaceutically acceptable form thereof, is administered to the subject at a dose of 0.5 mg to 20 mg per day or 0.5 mg to 10 mg per day.11 . The method of any one of claims 1-10, wherein the Compound 1 , or a pharmaceutically acceptable form thereof, is administered to the subject once or twice per day; optionally once or twice per day intermittently; optionally on a 7 days on / 7 days off schedule; optionally during a 28-day treatment cycle on days 1 to 7, 8 to 14, 15 to 21, 21 to 28, 1 to 7 and 15 to 21, 8 to 14 and 22 to 28, 1 to 21, or 1 to 28 of the treatment cycle.

12. The method of any one of claims 1-7, wherein the Compound 1, or a pharmaceutically acceptable form thereof, is administered intravitreally to the subject.

13. The method of claim 12, wherein the Compound 1, or a pharmaceutically acceptable form thereof, is administered to the subject at a dose of 0.005 mg / 0.05 mb to 4 mg / 0.10 mL per day.

14. The method of any one of claims 1-13, wherein the subject is a VEGF / VEGFR signaling pathway inhibitor-naive subject.

15. The method of any one of claims 1-13, wherein the subject has been treated previously with a VEGF / VEGFR signaling pathway inhibitor, for example, the subject has been treated previously with a VEGF / VEGFR signaling pathway inhibitor and is not currently being treated with a VEGF / VEGFR signaling pathway inhibitor, or the subject is currently being treated with a VEGF / VEGFR signaling pathway inhibitor.

16. The method of any one of claims 1-15, wherein the Compound 1, or a pharmaceutically acceptable form thereof, is a pharmaceutically acceptable salt of the Compound 1.

17. The method of claim 16, wherein the pharmaceutically acceptable salt is a benzoate salt, a besylate salt, a chloride salt, a citrate salt, a fumarate salt, a gentisate salt, a glycolate salt, a 1- hydroxy-2-naphthoate salt, a malate salt, a maleate salt, a mesylate salt, an oxalate salt, a phosphate salt, a tartrate salt, or a tosylate salt of the Compound 1.

18. The method of any one of claims 1-15, wherein the Compound 1, or a pharmaceuticallyacceptable form thereof, is a free base of the Compound 1 .

19. The method of any one of claims 1-15 or 18, wherein the Compound 1, or a pharmaceutically acceptable form thereof, is a pharmaceutically acceptable solvate of the Compound 1, or a pharmaceutically acceptable form thereof.

20. The method of claim 19, wherein the pharmaceutically acceptable solvate is selected from the group consisting of: a hydrate, a hemi-hydrate, an iso-butyl acetate solvate, an isopropyl acetate solvate, a tetrahydrofuran solvate, an acetone solvate, an acetonitrile solvate, or combinations thereof.

21. The method of any one of claims 1-15 or 18, wherein the Compound 1, or a pharmaceutically acceptable form thereof, is a non-solvate of the Compound 1 or a pharmaceutically acceptable salt thereof.

22. The method of any one of claims 1-15 or 18-20, wherein the Compound 1, or a pharmaceutically acceptable form thereof, is a crystalline, free base, hemi-hydrate of the Compound 1.

23. The method of any one of claims 1-15, 18-20, or 22, wherein the Compound 1, or a pharmaceutically acceptable form thereof, is Compound 1 (Form 1).

24. The method of claim 23, wherein the Compound 1 (Form 1) is characterized by an XRPD pattern, when measured using Cu Ka radiation, comprising peaks at approximately 9.0, 12.8, 16.6, and 18.4° 20; optionally wherein the XRPD pattern further comprises peaks at approximately 8.6, 12.0, 18.1, and 23.2° 29; optionally wherein the XRPD pattern further comprises peaks at approximately 16.1, 17.1, 24.1, and 25.6° 20.

25. A pharmaceutical composition comprising (a) Compound 1, or a pharmaceutically acceptable form thereof; and (b) a VEGF / VEGFR signaling pathway inhibitor.

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