Inhibitors of lysyl oxidase

Abstract

In general, disclosed herein are lysyl oxidase (LOX) family inhibitors of Formula (A): or a pharmaceutically acceptable salt thereof. Additionally, disclosed herein are methods directed to treating a disease of condition associated with the modulation or inhibition of lysyl oxidase (LOX).

Description

Patent Application Attorney Docket No.: USC-838-PCT (1726)INHIBITORS OF LYSYL OXIDASE CROSS-REFERENCE TO RELATED APPLICATION

[0001] The present application is related and has right of priority to U. S. Provisional Application No. 63 / 720,467 filed on November 14, 2024 in the United States Patent and Trademark Office and which is incorporated by reference in its entirety herein.GOVERNMENT SUPPORT CLAUSE

[0001] This invention was made with government support under R42 CA275622 and R01 CA267101 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND

[0002] Lysyl oxidase (LOX) is the founding member of the LOX family proteins and mediates the conversion of lysine residues in collagen I and elastin precursors into highly reactive aldehydes. LOX triggers crosslinking and stabilization of ECM proteins and regulates cell adhesion, motility, and invasion in a copper-dependent manner. LOX is upregulated in aggressive tumors and correlates with metastasis and worse clinical outcomes. Furthermore, the key role of LOX was also shown in therapy resistance. Given the multifaceted functions of LOX in cancer, there is a huge interest in identifying potent and tolerable LOX family inhibitors with minimum effects on other monoamine oxidases (MAOs). BAPN is the earliest and most commonly used research tool to inhibit LOX; however, it is not amenable to further optimization, inhibits all LOX family members and showed toxicity in the clinic. However, there are currently no potent, safe, and orally bioavailable small molecule inhibitors to treat desmoplastic solid tumors in clinics, thus necessitating the development of small molecule inhibitors to be tested in clinics to treat tumors driven by LOX family proteins.

[0003] Accordingly, alternative approaches and effective LOX family inhibitors (LOXi) are needed in the art.SUMMARY

[0004] In general, disclosed herein are lysyl oxidase (LOX) family inhibitors of Formula I:Patent Application Attorney Docket No.: USC-838-PCT (1726)R1or a pharmaceutically acceptable salt thereof, wherein:R4R5 ring A is aryl, heteroaryl.. or R1 through R3 are independently selected from hydrogen, -Ci-8 alkyl, -Ci-s alkoxy, or -OH;R4, R6, and R7 are independently selected from hydrogen, carboxyl, alkyl, alkyl ester, hydroxyl, methoxy, halogen, sulfonamide, phosphate, nitro, methylamine, boromc acid, -NH(C=O)CH?, -CO2Et, -CH=CH, -CH2-CH2, NH(C=O)-Ph-NH(C=O)CH?, -NH(C=O)-Ph-2,4-difluoro, -N2, N2'-(l,4-phenylene)bis(5'-methyl-[4,4'-bithiazole]-2,2'-diamine, -NH(C=O)CH2CH2OCH.3, -5’-methyl-[4,4’-bithiazole]-2,2’-diamine, -(C=O)NH- 3-F-Ph. -(C=O)N, N-di-CH3, -NH(C=O)CH2CH3CH2CH3. -NH(C=O)-Ph. -NH(C=O)CH2-4ClPh, -NH(C=O)-2.4-di-F-Ph, -NH(C=O)CH2-4-F-Ph. -(C=O)NH-Ph, -NH(C=0)-4-OMe-Ph, -(C=O)NH-Ph-NH(C=O)CH3, -(4-methylpiperazin-l-yl) methanone, -NH(C=O)- 4-pyridyl, -(C=O)NH-4-pyridyl, or -pyrimidinyl (3,5);R5 may be hydrogen; andX may be carbon or nitrogen.

[0005] Also, disclosed herein is disclosed herein is a compound of Formula (A):or a pharmaceutically acceptable salt thereof, wherein:Patent Application Attorney Docket No.: USC-838-PCT (1726)X is CH orN;R1is Ci-8 alkyl or Ci-8 haloalkyl;R2and R3are independently hydrogen, Ci-8 alkyl, Ci-8 alkylamino, -CH2-phenyl, or -C(O)Ra; wherein each Rais independently Ci-8 alkyl or phenyl;R4, R5, R6, and R7are each independently hydrogen, halogen, Ci-8 alkyl, Ci-8 alkylamino, Ci-8 alkoxy. -ORb, -N(Rb)2, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc.or R4and R5or R6and R7combine with the intervening atoms to form a phenyl, 5 to 6-membered heteroaryl, or 5 to 6-membered heterocycloalky 1 which is saturated or partially unsaturated, wherein the phenyl, heteroaryl and heterocycloalkyl are optionally substituted with one or more R8;each R8is independently halogen, Ci-8 alkyl. -ORb, oxo (=0), -N(Rb)2, -C(O)ORb, -C(O)RC, -C(0)N(Rb)2, or -NRbC(0)Rc;each Rbis independently hydrogen, Ci-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd;or two Rbcombine with the nitrogen atom to which they are attached to forms a 4 to 8-membered heterocycloalkyl which is optionally substituted with one or more Rd;each Rcis independently Ci-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; andeach Rdis independently halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), Ci-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, wherein the phenyl or heteroaryl is optionally substituted with one or more halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), -CH3, -C(0)0H, -C(O)OCH3, or -NHC(0)CH3.

[0006] Additionally, disclosed herein are methods directed to treating a disease of condition associated with the modulation or inhibition of lysyl oxidase (LOX). The method includes administering to a subject in need thereof a therapeutically effective amount of a LOX family inhibitor, or its pharmaceutically acceptable salt, disclosed herein.

[0007] Other features and aspects of the present disclosure are discussed in greater detail below.Patent Application Attorney Docket No.: USC-838-PCT (1726)BRIEF DESCRIPTION OF THE DRAWINGS

[0008] A full and enabling disclosure of the present disclosure is set forth more particularly in the remainder of the specification, including reference to the accompanying figures, in which:

[0009] FIG. 1A illustrates LOX family inhibitor discovery' pipeline.

[0010] FIG. IB depicts the percentage remaining cellular lysyl oxidase activity upon treatment of MDA-MB-231 cells with 5, 760 compounds for 48 hours.

[0011] FIG. 1C depicts inhibition of cell-based LOX activity and cell viability upon treatment with top 20 candidate LOX family inhibitors at 10 pM.

[0012] FIG. ID depicts doxorubicin sensitization upon combination of the selected 5 compounds (5 pM) with doxorubicin (1 pM) (n=4, 5 for vehicle, n=3 for treated).

[0013] FIG. IE depicts lysyl oxidase activity assay with the recombinant LOX protein incubated with 10 pM of 1, 2, 3, 4 and 5, and 10 mM BAPN. (pM = microM). Data represents mean values ± standard deviation SD). P-values were calculated with the unpaired, two-tailed Student’s t test. (* <0.05; ** P<0.01).

[0014] FIG. IF depicts lysyl oxidase activity assay with the recombinant LOXL2 protein incubated with 10 pM of 1, 2, 3, 4 and 5, and 10 mM BAPN. (pM = microM). Data represents mean values ± standard deviation SD). P- values were calculated with the unpaired, two-tailed Student’s t test. (* P<0.05, ** P<0.01).

[0015] FIG. 1G illustrates the scheme of assays and tested compounds in each assay together with compound 6403.

[0016] FIG. 1H depicts core structures of Formulas 1-1, 1-2, and 1-3.

[0017] FIG. II illustrates scheme of assays for testing novel derivatives and their selection flow in comparison to the parental compound 6403 and other two leads (6512 and 6487).

[0018] FIG. 2A depicts dose-dependent inhibition of recombinant LOX protein upon incubation with increasing doses of LXG6403.

[0019] FIG. 2B depicts dose-dependent inhibition of recombinant LOXL1 protein upon incubation with increasing doses of LXG6403.

[0020] FIG. 2C depicts dose-dependent inhibition of recombinant LOXL2 protein upon incubation with increasing doses of LXG6403.

[0021] FIG. 2D depicts dose response curves of LOX and LOXL2 activity upon incubation with increasing doses of LXG6403.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0022] FIG. 2E depicts dose dependent inhibition of concentrated LOX (rLOX-CM) upon incubation with increasing doses of LXG6403 or the LOXL2 inhibitor PAT-1251.

[0023] FIG. 2F depicts dose dependent inhibition of concentrated LOXL2 (rLOXL2-CM) upon incubation with increasing doses of LXG6403 or the LOXL2 inhibitor PAT-1251.

[0024] FIG. 2G depicts dose response curves of rLOX-CM and rLOXL2-CM activity’ upon incubation with increasing doses of LXG6403.

[0025] FIG. 2H depicts time- and concentration-dependent irreversible inhibition by LXG6403. Relative recovery’ of enzymatic activity' after pre-incubating rLOXL2 with 10X, 30X and 50X IC50 of LXG6403 for 30, 60 or 120 min.

[0026] FIG. 21 depicts substrate competition assay’ with rLOX-CM incubated with increasing doses of LXG6403 at different substrate concentrations. Data represents mean values ± standard deviation (SD) (n=2). P-values were calculated with the unpaired, two-tailed Student’s t test. (* F<0.05; ** P<0.01).

[0027] FIG. 2J depicts substrate competition assay with rLOXL2-CM incubated with increasing doses of LXG6403 at different substrate concentrations. Data represents mean values ± standard deviation (SD) (n=2). P-values were calculated with the unpaired, two-tailed Student’s t test. (* <0.05; ** F<0.01).

[0028] FIG. 3A depicts percentage cellular lysyl oxidase activity in LXG6403-treated MDA-MB-231 cells (n=3).

[0029] FIG. 3B depicts percentage cellular lysyl oxidase activity in LXG6403-treated HCC1143 cells (n=3).

[0030] FIG. 3C depicts percentage cellular lysyl oxidase activity in LXG6403-treated HCC 1937 cells (n=3).

[0031] FIG. 3D depicts percentage cellular lysyl oxidase activity in LXG6403-treated Hs-578-T cells (n=3).

[0032] FIG. 3E depicts DARTS assay showing the binding of LXG6403 to cellular LOX proteins. LXG6403 doses are written in pM. Dashed lines are added to separate different pronase dilutions.

[0033] FIG. 3F depicts quantification of band intensities from FIG. 3E at 1:5000 pronase dilutions. The bands of LOX, LOXL1 and LOXL2 were normalized to input (i.e., no pronase), as w ell as to the non-targeted protein controls with similar sizes as the LOX proteins (cyclin Bl for LOX and LOXL1, and vinculin for LOXL2) (n=2).Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0034] FIG. 3G depicts quantification of band intensities from E at 1:2000 pronase dilutions. The bands of LOX, LOXL1 and LOXL2 were normalized to input (i.e., no pronase), as well as to the non-targeted protein controls with similar sizes as the LOX proteins (cyclin Bl for LOX and LOXL1, and vinculin for LOXL2) (n=2).

[0035] FIG. 3H depicts percent grow th inhibition upon 48 hours of doxorubicin (1 pM) treatment in combination with 15 pM of LXG6403 in MDA-MB-231 cells (n=3-5 for vehicle, n=3 for treated).

[0036] FIG. 31 depicts percent growth inhibition upon 48 hours of cisplatin (40 pM) treatment in combination with 15 pM of LXG6403 in MDA-MB-231 cells (n=3-5 for vehicle, n=3 for treated).

[0037] FIG. 3J depicts percent growth inhibition upon 48 hours of paclitaxel (20 nM) treatment in combination with 15 pM of LXG6403 in MDA-MB-231 cells (n=3-5 for vehicle, n=3 for treated).

[0038] FIG. 3K depicts percent growth inhibition upon 48 hours of doxorubicin (1 pM) treatment in combination with 15 pM of LXG6403 in HCC1143 cells (n=3-5 for vehicle, n=3 for treated).

[0039] FIG. 3L depicts percent growth inhibition upon 48 hours of cisplatin (40 pM) treatment in combination with 15 pM of LXG6403 in HCC1143 cells (n=3-5 for vehicle, n=3 for treated).

[0040] FIG. 3M depicts percent growth inhibition upon 48 hours of paclitaxel (20 nM) treatment in combination with 15 pM of LXG6403 in HCC1143 cells (n=3-5 for vehicle, n=3 for treated).

[0041] FIG. 3N depicts percent growth inhibition of TNBC PDX organoids upon 9 days treatment with paclitaxel, cisplatin or doxorubicin with or without LXG6403. Data represents mean values ± standard deviation (SD) (n=3). (* P<0.05; ** <0.01; n.s. not significant).

[0042] FIG. 30 illustrates representative images of TNBC PDX organoids upon 9 days treatment with paclitaxel, cisplatin or doxorubicin w ith or w ithout LXG6403. Scale bar = 100 pm.

[0043] FIG. 4A depicts collagen I / Fibronectin staining in ECM derived from HFF-1 cells treated with vehicle or 15 pM of LXG6403 (n=2). Scale bar = 50 pm.

[0044] FIG. 4B depicts insoluble collagen assay in ECM derived from HFF-1 cells treated with vehicle or 15 pM of LXG6403 (n=2).Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0045] FIG. 4C depicts collagen I / Fibronectin staining in decellularized ECM (dECM) incubated with HCC1143 cells treated with 10 pM of 1 or LXG6403 (n=2). Scale bar = 50 pm.

[0046] FIG. 4D depicts insoluble collagen assay (D) in decellularized ECM (dECM) incubated with HCC1143 cells treated with 10 pM of 1 or LXG6403 (n=2).

[0047] FIG. 4E depicts relative doxorubicin autofluorescence in MDA-MB-231 (n=3) cells (n=2 for vehicle and LXG6403, and n=4 for doxorubicin and doxorubicin+LXG6403) embedded in collagen I and treated with the LOX inhibitor.

[0048] FIG. 4F depicts relative doxorubicin autofluorescence in HCC1143 (n=3) cells (n=2 for vehicle and LXG6403, and n=4 for doxorubicin and doxorubicin+LXG6403) embedded in collagen I and treated with the LOX inhibitor.

[0049] FIG. 4G depicts relative doxorubicin autofluorescence in TNBC organoids (n=2 for vehicle and LXG6403, and n=4 for doxorubicin and doxorubicin+LXG6403) embedded in collagen I and treated with the LOX inhibitor.

[0050] FIG. 4H depicts relative mitochondrial ROS levels in MDA-MB-231 cells treated with cisplatin with or without LXG6403 (n=4).

[0051] FIG. 41 depicts western blot analysis in MDA-MB-231 cells treated with cisplatin or doxorubicin with or without LXG6403. Actin was used as the loading control. Data represents mean values ± standard deviation (SD). F’-values were calculated with the unpaired, two-tailed Student’s t test. (* P<0.05 ** F’O. Ol).

[0052] FIG. 5A depicts tumor growth in TM01278 TNBC PDX model treated with doxorubicin (2 mg / kg, I. V, weekly), LXG6403 (50 mg / kg, P. O., daily) or their combination (n=6 for vehicle, Doxo, and LXG6403; n=5 for Combo).

[0053] FIG. 5B depicts tumor weights from mice in FIG. 5A.

[0054] FIG. 5C depicts quantification of Picrosirius Red staining in tumors from FIG.5A (n=15).

[0055] FIG. 5D illustrates representative images of Picrosirius Red staining in tumors from FIG. 5A (n=15). Scale bar = 100 pm.

[0056] FIG. 5E depicts insoluble collagen content of the tumors from FIG. 5A (n=3 different tumors with 2 replicates).

[0057] FIG. 5F depicts quantification of MP-SHG microscopy imaging from mice in FIG. 5A.

[0058] FIG. 5G illustrates representative images of MP-SHG microscopy imaging from mice in FIG. 5A.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0059] FIG. 5H depicts quantification of doxorubicin autofluorescence in tumors from FIG. 5A (n=3 different tumors with 2 replicates).

[0060] FIG. 51 illustrates representative images of doxorubicin autofluorescence in tumors from FIG. 5A (n=3 different tumors with 2 replicates). Scale bar = 500 pm.

[0061] FIG. 5J depicts quantification of DCFDA staining to show ROS levels in from FIG. 5A (n=3 different tumors with 2 replicates).

[0062] FIG. 5K illustrates representative images of DCFDA staining to show ROS levels in from FIG. 5A (n=3 different tumors with 2 replicates). Scale bar = 500 pm.

[0063] FIG. 5L depicts western blot analysis in tumors from FIG. 5A. Actin was used as the loading control.

[0064] FIG. 6A illustrates schematic summary of quantitative pharmacodynamics approach to characterize the global alterations in the ECM composition and organization upon targeting LOX with LXG6403.

[0065] FIG. 6B depicts heatmap of proteins identified by the ECM-targeted proteomics, showing differential expression of ECM proteins in different treatment groups of TNBC PDXs. *significant when comparing LXG6403 -treated tumors to tumors without LXG6403 treatment; f significant when comparing Doxo+ LXG6403 group to vehicle. The protein / peptide expression values are mapped to colors using the minimum and maximum of each row independently, setting the minimum to 0 and maximum to 1 for all protein / peptides.

[0066] FIG. 6C depicts image segmentation analysis of the MALDI-MSI data in TNBC PDXs under different treatment conditions. Data represents peptides (majorly collagen peptides and smaller fraction of non-collagen ECM proteins) heuristically clustered by spatial location and intensity. Clustering was done using the bisecting k-means method with Manhattan metric.

[0067] FIG. 6D depicts a heat map showing the abundance and PTMs of collagen peptides in TM01278 PDX tumors treated with doxorubicin in combination with LXG6403. The protein / peptide expression values are mapped to colors using the minimum and maximum of each row independently, setting the minimum to 0 and maximum to 1 for all protein / peptides.

[0068] FIG. 6E depicts selected modified fibrillar collagen peptides matched by high mass accuracy to sequence information from the same tissues as in C. Peptide modifications are reported with site probability > 95%. Significance among treatmentPatent Application Attorney Docket No.: USC-838-PCT (1726)groups was calculated using unpaired two-tailed student’s t-test (n=6). (* <0.05; ** P<0.01; *** P<0.001).

[0069] FIG. 7 illustrates characteristics of exemplary LOX family inhibitors.

[0070] FIG. 8A depicts relative activity inhibition of commercially available recombinant LOX, LOXL1 and LOXL2 proteins upon treatment with 0.5 mM BAPN. Data represents mean values ± standard deviation (SD) (n=2). P- values were calculated with the unpaired, two-tailed Student’s t test. (** P<0.01 ).

[0071] FIG. 8B depicts western blot showing the concentrated LOX and LOXL2 proteins in the conditioned media of HEK293T cells transfected with HA-tagged vectors of LOX and LOXL2.

[0072] FIG. 8C depicts prediction of LXG6403 binding site on LOXL2 using limited proteolysis assay and the AlphaFold-derived LOXL2 structure.

[0073] FIG. 8D depicts western blot analysis of LOX (NB100-2527) in TNBC cell lines. Actin was used as the loading control.

[0074] FIG. 9A depicts western blot analysis of LOX (L4794) in sgLOX-expressing MDA-MB-231 cells.

[0075] FIG. 9B depicts percent growth inhibition upon 48 hours of doxorubicin (1 pM) treatment in MDA-MB-231 cells after knocking out endogenous LOX and reconstitution with LOX ORF. Data represent mean values ± standard deviation (SD) (n=2-3). -values were calculated with unpaired, two-tailed Student’s t test. (* P<0.05 ** O. Ol).

[0076] FIG. 9C depicts percent growth inhibition upon 48 hours of cisplatin (40 pM) treatment in MDA-MB-231 cells after knocking out endogenous LOX and reconstitution with LOX ORF. Data represent mean values ± standard deviation (SD) (n=2-3). / ’-values were calculated with unpaired, two-tailed Student’s t test. (* <0.05; ** P<0.01).

[0077] FIG. 9D depicts percent growth inhibition upon 48 hours of paclitaxel (20 nM) treatment in MDA-MB-231 cells after knocking out endogenous LOX and reconstitution with LOX ORF. Data represent mean values ± standard deviation (SD) (n=2-3). -values were calculated with unpaired, two-tailed Student’s t test. (* PO.05 ** P<0.01).

[0078] FIG. 9E depicts real-time migration assay (RTCA) using MDA-MB-231 cells treated with 20pM of LXG6403 for 48 hours. Data represent mean values ± standard deviation (SD) (n=2-3). P-values were calculated with paired (E), two-tailed Student’s t test. (* P<0.05; ** ^<0.01).

[0079] FIG. 10A depicts western blot analysis of LOX (L4794) in siLOX-expressing MDA-MB-231 cells.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0080] FIG. 10B depicts relative mitochondrial ROS levels in MDA-MB-231 cells transfected with siLOX and treated with cisplatin. Data represent mean values ±SD (n=4). P-values were calculated with the unpaired, two-tailed Student’s t test. (** O. OI).

[0081] FIG. IOC depicts western blot analysis in MDA-MB-231 cells transfected with siLOX and treated with cisplatin. Actin was used as the loading control.

[0082] FIG. HA depicts cell viability assay in human endothelial cell line, HUVEC, treated with compound 1. Data represent mean values ±SD (n=4).

[0083] FIG. 1 IB depicts cell viability assay in human foreskin fibroblast cell line, HFF, treated with compound 1. Data represent mean values ±SD (n=4).

[0084] FIG. 11C depicts cell viability assay in normal breast cell line, MCF12A, treated with compound 1. Data represent mean values ±SD (n=4).

[0085] FIG. HD depicts cell viability assay in human endothelial cell line, HUVEC, human foreskin fibroblast cell line treated with LXG6403. Data represent mean values ±SD (n=4).

[0086] FIG. 1 IE depicts cell viability assay in human foreskin fibroblast cell line, HFF, treated with LXG6403. Data represent mean values ±SD (n=4).

[0087] FIG. 11 F depicts cell viability assay in normal breast cell line, MCF12A, treated with LXG6403. Data represent mean values ±SD (n=4).

[0088] FIG. 12A depicts body weight of mice treated with vehicle or 25, 50 or 75 mg / kg (P. O.) of LXG6403 for 5 days.

[0089] FIG. 12B depicts red blood cell (RBC) count of the mice from FIG. 1 1 A at Day 0 and Day 5.

[0090] FIG. 12C depicts count of lymphocytes (LYM) in blood of the mice from FIG.11 A at Day 0 and Day 5.

[0091] FIG. 12D depicts platelet (PLAT) count of the mice from FIG. 11 A at Day 0 and Day 5.

[0092] FIG. 12E depicts white blood cell (WBC) count of the mice from FIG. 11A at Day 0 and Day 5.

[0093] FIG. 12F depicts neutrophils (NEU) in blood count of the mice from FIG. 11 A at Day 0 and Day 5.

[0094] FIG. 12G depicts hematoxylin and eosin (H& E) staining of organs of BALB / c mice treated daily with 50 or 200 mg / kg of LXG6403 for 5 days.

[0095] FIG. 12H depicts percentage body weight change of mice bearing TM01278 PDX tumors and treated with doxorubicin alone or in combination with LXG6403. DataPatent Application Attorney Docket No.: USC-838-PCT (1726)represents mean values ± standard error of mean (SEM), and significance was calculated by Two way ANOVA for data in FIG. 12H and mean values ± standard deviation (SD) for data in FIGs. 12B-G(n=4-6).

[0096] FIG. 13A depicts subcellular localization of proteins found by targeted extracellular matrix proteomics upon treatment of TNBC PDXs with LXG6403 alone or in combination with chemotherapy. Green represents the proteins involved in the pathway and black represents proteins that are not involved in the pathway.

[0097] FIG. 13B depicts DAVID pathways analysis of differentially expressed proteins upon treatment of TNBC PDXs with LXG6403 alone or in combination with chemotherapy. Green represents the proteins involved in the pathway and black represents proteins that are not involved in the pathway.

[0098] FIG. 14 illustrates general scheme to synthesize LOX family inhibitors.

[0099] FIG. 15 A depicts percent cellular LOX activity in MDA-MB-231 TNBC cells treated with LOX family inhibitor 6403 for 48 hrs.

[0100] FIG. 15B depicts percent cellular LOX activity in MDA-MB-231 TNBC cells treated with LOX family inhibitor 6506 for 48 hrs.

[0101] FIG. 15C depicts percent cellular LOX activity in MDA-MB-231 TNBC cells treated with LOX family inhibitor 6532 for 48 hrs.

[0102] FIG. 16A depicts percent cell viability of human foreskin fibroblast (HFF) and human embryonic kidney (HEK293T) cells treated with increasing doses of LOX family inhibitor 6403 for 48 hrs.

[0103] FIG. 16B depicts percent cell viability of human foreskin fibroblast (HFF) and human embryonic kidney (HEK293T) cells treated with increasing doses of LOX family inhibitor 6506 for 48 hrs.

[0104] FIG. 16C depicts percent cell viability of human foreskin fibroblast (HFF) and human embryonic kidney (HEK293T) cells treated with increasing doses of LOX family inhibitor 6532 for 48 hrs.

[0105] FIG. 17A depicts percentage of LOX activity upon incubation with LOX family inhibitor 6403.

[0106] FIG. 17B depicts percentage of LOX activity upon incubation with LOX family inhibitor 6487.

[0107] FIG. 17C depicts percentage of LOX activity upon incubation with LOX family inhibitor 6505.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0108] FIG. 17D depicts percentage of LOX activity upon incubation with LOX family inhibitor 6512.

[0109] FIG. 17E depicts percentage of LOXL2 activity upon incubation with LOX family inhibitor 6403.

[0110] FIG. 17F depicts percentage of LOXL2 activity upon incubation with LOX family inhibitor 6487.

[0111] FIG. 17G depicts percentage of LOXL2 activity upon incubation with LOX family inhibitor 6505.

[0112] FIG. 17H depicts percentage of LOXL2 activity upon incubation with LOX family inhibitor 6512.

[0113] FIG. 18 depicts relative LOXL1 activity upon incubation with LOXi at 2 pM.

[0114] FIG. 19A depicts the protection of HA-tagged LOX, LOXL1 and LOXL2 from pronase digestion (1:2000 for LOX and LOXL1, 1:3500 for LOXL2) upon treatment with 20 pM of LOXi for LOX and LOXL2. and 50 pM LOXi for LOXL1 detected with the HA tag antibody.

[0115] FIG. 19B depicts quantification of normalized band intensities. The bands of LOX were normalized to input (i.e., no pronase) as well as to the non-targeted protein controls with similar sizes as the LOX proteins (Cyclin B 1 for LOX).

[0116] FIG. 19C depicts quantification of normalized band intensities. The bands of LOXLlbwere normalized to input (i.e., no pronase) as well as to the non-targeted protein controls with similar sizes as the LOX proteins (Cyclin Bl for LOXL).

[0117] FIG. 19D depicts quantification of normalized band intensities. The bands of LOXL2 were normalized to input (i.e., no pronase) as well as to the non-targeted protein controls with similar sizes as the LOX proteins (vinculin for LOXL2).

[0118] FIG. 20A depicts combination indices heatmaps showing the synergistic cell viability inhibition in collagen I-embedded MDA-MB-231 cells pretreated with LOXi for 48 hrs followed by 48 hrs treatment with doxorubicin or paclitaxel.

[0119] FIG. 20B depicts combination indices heatmap for collagen I-embedded T47D cells pretreated with LOXi for 48 hrs followed by 48 hrs treatment with doxorubicin.

[0120] FIG. 21 illustrates representative images of the combination indices (left) and heatmaps (right) from FIG. 20B.

[0121] FIG. 22 illustrates Collagen / Fibronectin staining in MDA-MB-231 cells cultured on HFF-derived ECM and treated with LOXi for 48 hrs.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0122] FIG. 23 A depicts Collagen I+matrigel-embedded MDA-MB-231 cells pretreated with LOXi (35 pM) for 24 hrs followed by the addition of doxorubicin (1 pM) for 24 hrs.

[0123] FIG. 23B-D depicts quantification of the band intensities of p-FAK (Y397) relative to control.

[0124] FIG. 23C depicts quantification of the band intensities of p-Src (Y416) relative to control.

[0125] FIG. 23D depicts quantification of the band intensities of p-H2AX (SI 39) relative to control.

[0126] FIG. 24A depicts MAO-A activity upon incubation with LOXi. The IC50s are provided on the graphs.

[0127] FIG. 24B depicts MAO-B activity upon incubation with LOXi. The IC50s are provided on the graphs.

[0128] FIG. 25A depicts tumor growth in TM00099 TNBC PDX model treated with cyclophosphamide (40 mg / kg, I. V, weekly), LXG6403 (50 mg / kg, P. O., bidaily) or their combination. Data represents mean values ±SD for the bar graph, while it represents mean values ± standard error of the mean (SEM) for the tumor volume graph. Significance for the tumor volume graph was calculated by two-way ANOVA. (** O. Ol).

[0129] FIG. 25B depicts Pharmacodynamic (PD) analysis showing reduced lysyl oxidase activity in TM00099 PDX tumors upon LXG6403 treatment in vivo. Data represents mean values ±SD for the box plots, while it represents mean values ± standard error of the mean (SEM) for the tumor volume graph. Significance for the tumor volume graph was calculated by two-way ANOVA. (** / NO.01).

[0130] FIG. 25C depicts percentage body weight change of mice bearing TM00099 PDX tumors from FIG. 25A. Data represents mean values ±SD for the bar graph, while it represents mean values ± standard error of the mean (SEM) for the tumor volume graph. Significance for the tumor volume graph was calculated by two-way ANOVA. (** <0.01).

[0131] FIG. 25D depicts count of neutrophils in blood of the mice from FIG. 25A at the end of the experiment at day 22. Data represents mean values ±SD for the bar graphs and box plots, while it represents mean values ± standard error of the mean (SEM) for the tumor volume graph. Significance for the tumor volume graph was calculated by two-way ANOVA. (** P<0.01).Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0132] FIG. 25E depicts count of platelets in blood of the mice from FIG. 25A at the end of the experiment at day 22. Data represents mean values ±SD for the bar graphs and box plots, while it represents mean values ± standard error of the mean (SEM) for the tumor volume graph. Significance for the tumor volume graph was calculated by two-way ANOVA. (** O. Ol).

[0133] FIG. 26A illustrates a schematic of the experimental details to test the effects of LXG6403 on spontaneous metastasis in transgenic MMTV-PyMT mice.

[0134] FIG. 26B depicts putative number of circulating tumor cells (CTCs) in MMTV-PyMT mice at 3 weeks age before treatment and 5 weeks age after 2 weeks of treatment with LXG6403 (50 mg / kg, P. O., daily).

[0135] FIG. 26C depicts percentage body weight change of MMTV-PyMT mice treated with LXG6403 (50 mg / kg, P. O., daily) for 64 days.

[0136] FIG. 26D depicts changes in tumor weights of the mice from FIG. 26C.

[0137] FIG. 26E depicts representative images of lung metastatic nodules in mice from FIG. 26C. Upper panel: Nodule counts. Lower panel: H& E stain.

[0138] FIG. 26F depicts percentage lung metastatic nodules in mice from FIG. 26C. Data represents mean values ±SD for box blots, while it represents mean values ± standard error of the mean (SEM) for the body weight graph. -values were calculated with the unpaired, two-tailed Student’s t test (* P<0.05).

[0139] FIG. 27A illustrates a schematic of the experimental details to test the effects of LXG6403 on anti-PD-Ll (PD-L1 mAh) response of transgenic MMTV-PyMT mice.

[0140] FIG. 27B depicts tumor growth in MMTV-PyMT mice treated with PD-L1 mAb (5 mg / kg, I P, biweekly), LXG6403 (50 mg / kg, P. O., bidaily) or their combination.

[0141] FIG. 27C depicts percentage body weight change of mice from FIG. 27B.

[0142] FIG. 27D depicts counts of lymphocytes in blood of the mice from FIG. 27B at the end of the experiment at day 28.

[0143] FIG. 27E depicts counts of monocytes in blood of the mice from FIG. 27B at the end of the experiment at day 28.

[0144] FIG. 27F depicts counts of neutrophils in blood of the mice from FIG. 27B at the end of the experiment at day 28.

[0145] FIG. 27G depicts counts of platelets in blood of the mice from FIG. 27B at the end of the experiment at day 28.

[0146] FIG. 28 depicts LOX inhibition enhances the effectiveness of standard-of-care in kidney cancer xenografts in vivo. Percentage change of tumor volume from baseline inPatent Application Attorney Docket No.: USC-838-PCT (1726)786-0 xenografts treated with Axitinib (30 mg / kg, P. O., bidaily), LXG6403 (75 mg / kg, P. O., bidaily) or their combination for 3 weeks.

[0147] FIG. 29 depicts the effect of representative LOXi on cell-based LOX activity. A-F. Normalized cellular LOX activity in MDA-MB-231 (A-C) and Hs578T (D-F) TNBC cells treated with parental compound 6403 (A), or novel analogs 6576 (B), and 6609 (C) for 48 hrs.

[0148] FIG. 30 depicts the effect of selected LOXi on MAO-A and -B activity. A, B. Percent MAO-A (A) or MAO-B (B) activity upon incubation with LOXi. The IC50s are provided at Table 3.

[0149] FIG. 31 depicts the effect of representative LOXi on normal (non-cancerous) cell viability. Relative cell viability of human umbilical vein endothelial cells (HUVEC) and mouse epithelial -like cells, NMuMG treated with escalated doses of parental compound 6403 (A), or novel analogs 6576 (B), and 6609 (C) for 48 hrs.

[0150] FIG. 32 depicts the effects of representative LOXi on recombinant LOX and LOXL2 activity. A-C. Percent LOX activity upon incubation with LOXi. D-F. Percent LOXL2 activity upon incubation with LOXi (summarized in Table 4).

[0151] FIG. 33 depicts the effects of selected LOXi on recombinant LOXL1 activity. Relative LOXL1 activity upon incubation with LOXi at 2 pM (summarized in Table 4).

[0152] FIG. 34 depicts DARTS assay showing the binding of LOXi to cellular LOX. LOXL2, or LOXL1 proteins. A. The protection of HA-tagged LOX, LOXL1 and LOXL2 from pronase digestion (1:2000 for LOX and LOXL1, 1:3500 for LOXL2) upon treatment with 20 pM of LOXi for LOX and LOXL2, and 50 pM LOXi for LOXL1 detected with the HA tag antibody. B-D. Quantification of normalized band intensities. The bands of LOX, LOXL1, and LOXL2 were normalized to input (i.e.. no pronase) as well as to the nontargeted protein controls with similar sizes as the LOX proteins (Cyclin Bl for LOX and LOXL1, and vinculin for LOXL2).

[0153] FIG. 35 depicts chemotherapy sensitization upon treatment of collagen I-embedded human TNBC cell line MDA-MB-231 and ER+ breast cancer T47D cells with selected LOXi. A, B. Combination indices heatmaps showing the synergistic cell viability inhibition in collagen I-embedded MDA-MB-231 cells pretreated with LOXi for 48 hrs followed by 48 hrs treatment with doxorubicin (A) or paclitaxel (B).

[0154] FIG. 36 depicts doxorubicin sensitization upon treatment of chemoresistant TM01278 TNBC PDX organoid model with selected LOXi. Heatmaps represent growth inhibition (left side, red scale bar 100 pm) and combination indices (right side).Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0155] FIG. 37 depicts LOXi-driven changes in fibrillar collagens. Insoluble collagen assay in MDA-MB-231 cells cultured on HFF-derived ECM and treated with LOXi for 48 hrs.

[0156] FIG. 38 depicts FAK / Src signaling inhibition and DNA damage induction upon combination of LOXi with chemotherapy. A. Collagen I+matrigel-embedded MDA-MB-231 cells were pretreated with LOXi (35 pM) for 24 hrs followed by addition of doxorubicin (1 pM) for 24 hrs. B-D. Quantification of the band intensities of p-FAK (Y397) (B), p-Src (Y416) (C) and p-H2AX (S139) (D) relative to control.

[0157] FIG. 39 depicts collagen / fibronectin staining in MDA-MB-231 cells. Cells were cultured on HFF-derived ECM and treated with indicated LOXi (20 pM) for 48 hrs and IF imaging was performed. Scale bar 100 pm.

[0158] FIG. 40 depicts KinomeScan and CEREP47 assays to evaluate potential off targets. A. Bar plot represents selectivity scores of three compounds. Selectivity score S(10) & S(l) are the ratio of number of non-mutant kinases with %Ctrl <10 and 1, over number of non-mutant kinases tested, respectively. For example, S(10) of 6512 is 0.022 (=9 / 403), suggesting that lower ratio indicates better selectivity toward LOX. B. Common off-targets of selective derivatives in CEREP47 SAFETY scan E / IC50. Three compounds hit HTR2B; however, their potency was 4 - 10 folds less than Ctrl (LY272015). All four compounds hit COX2; however, their potency was 4 - 6 folds less than Ctrl (NS398). Note: 6576 did not hit HTR2B.

[0159] Repeat use of reference characters in the present specification and drawings is intended to represent the same or analogous features or elements of the present invention.DETAILED DESCRIPTION

[0160] Reference will now be made in detail to various embodiments of the presently disclosed subject matter, one or more examples of which are set forth below. Each embodiment is provided by way of explanation, not limitation, of the subj ect matter. In fact, it will be apparent to those skilled in the art that various modifications and variations may be made to the present disclosure without departing from the scope or spirit of the disclosure. For instance, features illustrated or described as part of one embodiment may be used in another embodiment to yield a still further embodiment. Thus, it is intended that the present disclosure covers such modifications and variations as come within the scope of the appended claims and their equivalents.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0161] LOX / LOXLs are highly atractive therapeutic targets playing crucial roles in many cancer-associated processes, such as cancer cell invasion, angiogenesis, and therapy resistance. Despite having key roles in promoting cancer progression, there are currently no potent, safe, and orally bioavailable small molecule inhibitors of LOX family proteins to treat solid tumors in clinics. Current limitations of identifying LOX family inhibitors include the lack of 3D structure of LOX to perform structure-based drug discovery and the use of only cell-free screen-based approaches leading to inhibitors with limited cellular lysyl oxidase activity inhibition. To address these deficits, a robust high-throughput cellbased screening pipeline is disclosed herein and coupled it to SAR analysis and optimization that led to the discovery of chemically distinct LOX family inhibitors.

[0162] Surprisingly, LOX family inhibitors disclosed herein inhibited LOX activity and reduced ECM crosslinking, resulting in increased drug penetration, and led to chemosensitization in TNBC cells or organoids embedded in collagen without any cytotoxicity in normal cells. Integration of the state-of-the-art techniques demonstrated that LOX family inhibitors disclosed herein, such as LXG6403, reduce (1) fibrillar collagen content, as shown by picrosirius red staining and ECM-targeted proteomics and (2) collagen crosslinking, as shown by insoluble collagen assay and MP-SHG microscopy in chemoresistant TNBC PDXs. This further increase chemotherapy penetration, leading to FAK inhibition and activation of ROS / DNA damage axis that ultimately result in G1 arrest and apoptosis in vivo.

[0163] The LOX family consists of 5 different members (LOX, LOXL1, LOXL2, LOXL3 and LOXL4). Among all, LOX (first identified member) and LOXL2 are the two most oncogenic members that have been heavily associated with cancer progression, particularly with metastasis and recently with chemoresistance. LOX family proteins also have versatile and complex functions not only in cancer but also in fibrosis and other diseases upon interacting with many canonical and non-canonical partners. Being key regulators in various diseases, development of potent and safe inhibitors of LOX family proteins is of great interest.

[0164] In some example embodiment, a lysyl oxidase family inhibitor (LOXi) may have the general structure of Formula I:Patent Application Attorney Docket No.: USC-838-PCT (1726)or a pharmaceutically acceptable salt thereof, wherein:

[0165] ring A may be aryl, heteroar l.R4

[0166] R1 through R3 may be independently selected from hydrogen, -Ci-8 alkyl, -Ci-8 alkoxy, or -OH;

[0167] R4, R6, and R7 are independently selected from hydrogen, carboxyl, alkyl, alkyl ester, hydroxyl, methoxy, halogen, sulfonamide, phosphate, nitro, methylamine, boronic acid, -NH(C=O)CH3, -CO2Et, -CH=CH, -CH2-CH2, NH(C=O)-Ph-NH(C=O)CH3, -NH(C=O)-Ph-2.4-difluoro, -N2, N2'-(1.4-phenylene)bis(5'-methyl-[4,4'-bithiazole]-2,2'-diamine, -NH(C=O)CH2CH2OCH3, -5’-methyl-[4,4’-bithiazole]-2,2’-diamine, -(C=O)NH- 3-F-Ph, -(C=O)N, N-di-CH3, -NH(C=O)CH2CH3CH2CH3, -NH(C=O)-Ph, -NH(C=O)CH2-4ClPh, -NH(C=O)-2,4-di-F-Ph, -NH(C=O)CH2-4-F-Ph, -(C=O)NH-Ph, -NH(C=O)-4-OMe-Ph, -(C=O)NH-Ph-NH(C=O)CH3, -(4-methylpiperazin-l-yl) methanone, -NH(C=O)- 4-pyridyl, -(C=O)NH-4-pyridyl, or -pyrimidinyl (3,5);

[0168] R5 may be hydrogen; and

[0169] X may be carbon or nitrogen.

[0170] In some example embodiments, R1 may be independently selected from hydrogen, -Ci-8 alkyl, -Ci-8 alkoxy, or -OH. In one example embodiment, for instance. RI may be hydrogen. In another example embodiment, for instance, Rl may be -CH3.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0171] In some example embodiments, R2 may be independently selected from hydrogen, -Ci-8 alkyl, -Ci-8 alkoxy, or -OH. In one example embodiment, for instance. R2 may be hydrogen. In another example embodiment, for instance, R2 may be -COCH3.

[0172] In some example embodiments, R3 may be independently selected from hydrogen, -C1-8 alkyl, -C1-8 alkoxy, or -OH. In one example embodiment, for instance, R3 may be hydrogen. In another example embodiment, for instance, R3 may be -COCH3. In yet another example embodiment, for instance, R3 may be -CH3.

[0173] In some example embodiments, ring A may be aryl, heteroaryl,In one example embodiment, for instance, ring A may R5beA 'XxX R? ]nanother example embodiment, for instance, ring A may beR4AX

[0174] In some example embodiments, R4 may be independently selected from hydrogen, carboxyl, alkyl, alkyl ester, hydroxyl, methoxy, halogen, sulfonamide, phosphate, nitro, methylamine, boronic acid, -NH(C=O)CH3, -CChEt. -CH=CH, -CH2-CH2, NH(C=O)-Ph-NH(C=O)CH3, -NH(C=O)-Ph-2,4-difluoro, -N2, N2'-(1,4-phenylene)bis(5'-methyl-[4,4'-bithiazole]-2,2'-diamine, -NH(C=O)CH2CH2OCH3, -5’-methyl-[4,4’-bithiazole]-2,2’-diamine, -(C=O)NH-3-F-Ph, -(C=O)N, N-di-CH3, - NH(C=O)CH2CH3CH2CH3, -NH(C=O)-Ph, -NH(C=O)CH2-4ClPh, -NH(C=O)-2,4-di-F-Ph, -NH(C-O)CH2-4-F-Ph. -(C O)NH-Pli. -NH(C=O)-4-OMe-Ph, -(C O)NH-Ph- NH(C=O)CH3, -(4-methylpiperazin-l-yl) methanone, -NH(C=O)-4-pyridyl, -(C=O)NH-4-pyridyl, or -pyrimidinyl (3,5). In one example embodiment, for instance, R4 may be hydrogen. In another example embodiment, for instance, R4 may be -NH(C=O)CH3. In yet another example embodiment, for instance, R4 may be -CChEt.

[0175] In some example embodiments, R5 may be hydrogen.

[0176] In some example embodiments, R6 may be independently selected from hydrogen, carboxyl, alkyl, alkyl ester, hydroxyl, methoxy, halogen, sulfonamide, phosphate, nitro, methylamine, boronic acid, -NH(C=O)CH3, -CChEt, -CH=CH, -CH2-CH2,Patent Application Attorney Docket No.: USC-838-PCT (1726)NH(C=O)-Ph-NH(C=O)CH3, -NH(C=O)-Ph-2,4-difluoro, -N2, N2'-(1,4-phenylene)bis(5'-methyl-[4,4'-bithiazole]-2,2'-diamine, -NH(C=O)CH2CH2OCH3, -5’-methyl-[4,4'-bithiazole]-2,2’-diamine, -(C=0)NH-3-F-Ph, -(C=O)N, N-di-CH3, -NH(C=O)CH2CH3CH2CH3, -NH(C=O)-Ph, -NH(C=O)CH2-4ClPh, -NH(C=O)-2,4-di-F-Ph, -NH(C=O)CH2-4-F-Ph, -(C=O)NH-Ph, -NH(C=0)-4-0Me-Ph, -(C=O)NH-Ph-NH(C=O)CH3, -(4-methylpiperazin-l-yl) methanone, -NH(C=O)-4-pyridyl, -(C=O)NH-4-pyridyl, or -pyrimidinyl (3,5). In one example embodiment, for instance, R6 may be hydrogen. In another example embodiment, for instance, R6 may be -NH(C=O)CH3. In yet another example embodiment, for instance, R6 may be -NH(C=O)CH2CH2OCH3. In another example embodiment, for instance, R6 may be -5'-methyl-[4,4'-bithiazole]-2,2'-diamine. In another example embodiment, for instance, R6 may be -(C=O)NH-3-F-Ph. In another example embodiment, for instance, R6 may be -(C=O)N, N-di-CH3. In another example embodiment, for instance, R6 may be -NH(C=O)CH2CH3CH2CH3. In another example embodiment, for instance, R6 may be -NH(C=O)-Ph. In another example embodiment, for instance, R6 may be -NH(C=O)CH2-4ClPh. In another example embodiment, for instance, R6 may be -NH(C=O)CH2-4-F-Ph. In another example embodiment, for instance, R6 may be -COOH. In another example embodiment, for instance, R6 may be -(C=O)NH-Ph-NH(C=O)CH3. In another example embodiment, for instance, R6 may be -NH(C=0)-4-0Me-Ph. In another example embodiment, for instance, R6 may be -(4-methylpiperazin- 1 -yl) methanone. In another example embodiment, for instance, R6 may be -NH(C=O)-4-pyridyl. In another example embodiment, for instance, R6 may be -(C=O)NH-4-pyridyl. In another example embodiment, for instance, R6 may be pyrimidinyl (3,5).

[0177] In some example embodiments, R7 may be independently selected from hydrogen, carboxyl, alkyl, alkyl ester, hydroxyl, methoxy, halogen, sulfonamide, phosphate, nitro, methylamine, boronic acid, -NH(C=O)CH3, -CO2Et, -CH=CH, -CH2-CH2, -NH(C=O)-Ph-NH(C=O)CH3, -NH(C=O)-Ph-2,4-difluoro, -N2, N2'-(l,4-phenylene)bis(5'-methyl-[4,4'-bithiazole]-2,2'-diamine, -NH(C=O)CH2CH2OCH3, -5'-methyl-[4,4'-bithiazole]-2,2'-diamine, -(C=O)NH-3-F-Ph. -(C=O)N, N-di-CH3, -NH(C=O)CH2CH3CH2CH3, -NH(C=O)-Ph, -NH(C=O)CH2-4ClPh, -NH(C=O)-2,4-di-F-Ph, -NH(C=O)CH2-4-F-Ph, -(C=O)NH-Ph, -NH(C=O)-4-OMe-Ph, -(C=O)NH-Ph-NH(C=O)CH3, -(4-methylpiperazin-l-yl) methanone, -NH(C=O)-4-pyridyl, -(C=O)NH-4-pyridyl, or -pyrimidinyl (3,5). In one example embodiment, for instance, R7 may be hydrogen. In another example embodiment, for instance, R5 may be -NH(C=O)CH3. In yet another example embodiment, for instance,Patent Application Attorney Docket No.: USC-838-PCT (1726)R7 may be -CH=CH. In another example embodiment, for instance, R7 may be -CH2-CH2. In another example embodiment, for instance. R7 may be NH(C=O)-Ph-NH(C=O)CH3. In another example embodiment, for instance, R7 may be -NH(C=O)-Ph-2,4-difluoro. In another example embodiment, for instance, R7 may be -5'-methyl-[4,4'-bithiazole]-2,2'-diamine. In another example embodiment, for instance, R7 may be COOH. In another example embodiment, for instance, R7 may be -(C=O)NH-Ph. In another example embodiment, for instance, R7 may be -(C=O)NH-Ph-NH(C=O)CH3.

[0178] In some example embodiments, X may be carbon or nitrogen. In one example embodiment, for instance, X may be carbon. In another example embodiment, for instance, X may be nitrogen.

[0179] In some example embodiment, a lysyl oxidase family inhibitor (LOXi) may have the general structure of Formula (A):R1or a pharmaceutically acceptable salt thereof, wherein:

[0181] X is CH or N;

[0182] R1is Ci-8 alkyl or Ci-s haloalkyl;

[0183] R2and R3are independently hydrogen, Ci-8 alkyl, Ci-8 alkylamino, -CH2-phenyl, or -C(O)Ra; wherein each Rais independently Ci-s alkyl or phenyl;

[0184] R4, R5, R6, and R7are each independently hydrogen, halogen, C1-8 alkyl, C1-8 alkylamino, C1-8 alkoxy, -ORb, -N(Rb)2, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc,

[0185] or R4and R5or R6and R7combine with the intervening atoms to form a phenyl, 5 to 6-membered heteroaryl, or 5 to 6-membered heterocycloalkyl which is saturated or partially unsaturated, wherein the phenyl, heteroaryl and heterocycloalkyl are optionally substituted with one or more R8;

[0186] each R8is independently halogen, C1-8 alkyl, -ORb, oxo (=0), -N(Rb)2, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc;Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0187] each Rbis independently hydrogen, Ci-s alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd;

[0188] or two Rbcombine with the nitrogen atom to which they are attached to forms a 4 to 8-membered heterocycloalkyl which is optionally substituted with one or more Rd;

[0189] each Rcis independently Ci-s alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; and

[0190] each Rdis independently halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, wherein the phenyl or heteroaryl is optionally substituted with one or more halogen, -OH, -OCH3, -NH2, -NHCHs, -N(CH3), -CH3, -C(O)OH, -C(O)OCH3, or -NHC(O)CH3.R5Xf ^X^^R

[0191] In one example embodiment, when ring Ais '7, then at least one of R4, R5. R6. or R7is not hydrogen.

[0192] In one example embodiment, the lysyl oxidase family inhibitor of Formula I may be represented by the following structure:R5

[0193] or a pharmaceutically acceptable salt thereof.

[0194] In another example embodiment, the lysyl oxidase family inhibitor of Formula I may be represented by the following structure:Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0195] or a pharmaceutically acceptable salt thereof.

[0196] In another example embodiment, the lysyl oxidase family inhibitor of Formula I may be represented by the following structure:R1 R4(1-3),

[0197] or a pharmaceutically acceptable salt thereof, wherein:

[0198] R1is -CH3, -CH2CH3, or -CF3;

[0199] R2and R3are independently hydrogen, C1-8 alkyl, or -C(O)Ra; wherein each Rais independently C1-8 alkyl or phenyl;

[0200] R4is hydrogen, C1-8 alkyl, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc;

[0201] R5is hydrogen, C1-8 alkyl, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc;

[0202] each Rbis independently hydrogen, C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd;

[0203] or two Rbcombine together with the nitrogen atom to which they are attached to forms a 4 to 8-membered heterocycloalkyl which is optionally substituted with one or more Rd;

[0204] each Rcis independently C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; and

[0205] each Rdis independently halogen, -OH, -OCH3, -NH2, -NHCHy -N(CH3), C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, wherein the phenyl or heleroaryl is optionally substituted with one or more halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), -CH3, -C(O)OH. -C(O)OCH3, or -NHC(O)CH3.

[0206] In another example embodiment, the lysyl oxidase family inhibitor of Formula I may be represented by the following structure:

[0207] or a pharmaceutically acceptable salt thereof, w herein:Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0208] X is CH or N;

[0209] R1is -CHs, -CH2CH3, or -CF3;

[0210] R2and R3are independently hydrogen, C1-8 alkyl, or -C(O)Ra; wherein each Rais independently C1-8 alkyl or phenyl;

[0211] R4, R5, R6, and R7are each independently hy drogen, halogen, Ci-s alkyl, C1-8 alkylamino, C1-8 alkoxy. -ORb, -N(Rb)2, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc.

[0212] or R4and R5or R6and R7combine with the intervening atoms to form a phenyl, 5 to 6-membered heteroaryl, or 5 to 6-membered heterocycloalkyl which is saturated or partially unsaturated, wherein the phenyl, heteroaryl, and heterocycloalkyl is optionally substituted with one or more R8.

[0213] each R8is independently halogen, Ci-s alkyl, -ORb, oxo (=0), -N(Rb)2, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc;

[0214] each Rbis independently hydrogen, C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd;

[0215] or two Rbcombine with the nitrogen atom to which they are attached to forms a 4 to 8-membered heterocycloalkyl which is optionally substituted with one or more Rd;

[0216] each Rcis independently C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; and

[0217] each Rdis independently halogen, -OH, -OCH3, -NH2. -NHCH3, -N(CH3), C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, wherein the phenyl or heteroaryl is optionally substituted with one or more halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), -CH3, -C(O)OH, -C(O)OCH3, or -NHC(O)CH3;R5Rt J. R6

[0218] wherein when ring Ais, then at least one of R4, R5, R6, or R7is not hydrogen.

[0219] In some example embodiments, the lysyl oxidase family inhibitor of Formula I may be a compound having one of the following structures in Table A:Patent Application Attorney Docket No.: USC-838-PCT (1726)64156425 NC-A I Cl U2AsH NNH H6426 NCH [Pl Q JAs' NA'NA^ s ZT 'N' YH2XN Hb v / H IL z yo c o—’ / H HN'Y Yz_ N, N. / 6487A 7-~H2N x s (A -OC'<\ Y Y f Yz / Z0- <- — V-S AA 0| Cs zY z=A zX zx6488 NC — A CoH2N As' HHcoNX-A[ C6489 z — TOAsH2NbH Lb oz6505ovAN HH / H Ho N-4 N6506V H T V Y J H H6512 A frNn vH2N s Vs 06519-A6519-B o Avx JOYW AsA~-N 1 \ _H<3 ~6522Patent Application Attorney Docket No.: USC-838-PCT (1726) / 6523 NA— b i nH2N H6524H2N xkx. Hnsra,,6528AYH2N2 1 H9 x x,nH 0 F65H2N k H "ra,6530 NA / -O fj A H AlH2N As H06531NA- / 1 JTJOHH2N, HNA A J HA i I F6532 H2N s H \ / AASy> NH2N6533H2N H rf^H2K „V6538 VA N^NH206539H2N AsHPatent Application Attorney Docket No.: USC-838-PCT (1726)06554. A FS AN Y-X A A JH2N AsHHH6555 / u rr vN Y-x A ASJL U °T H2N AsHHz o6556 —fk A o.H2N Y z H / IZho6558 NVY \ Y nH2N As' ^' Y^ HAA °zz o A" NA z6559H2N A. H AIL / 0 ^cn zz N>6560NA-Y AAsOHNH2N |11 / X A IL J 9L z\6561 As N NVN Y Xi oH2N H H || 1 KH A H I6562 II / Y J<s. LL JL JLAsNN N X Y H2N Hh||65636568 NA AA rYiH2M As^ H APatent Application Attorney Docket No.: USC-838-PCT (1726)65696570 y z— / H2N H_ / xMzxNG _ / ^N^N L z^V06571H2N HHNr^.A Z= T1 1N < z HXNA-- / A6572 jG]H2N HHN^O6573N" V-< TIAS: HN^N^NH2N H6574 N V-O. 1 1 J1~S NH2N HJ6574-B I 1 1 1 H Is^ N^W y^H2N H IIJ6574-Cn^\— < i 1 T 1HH2N 1 IIs^\ S^NH20 HN N y-N6574-DH1 A J1 I 1 H6574-E H2N „ T TVN”SYN^<N^yLT0Patent Application Attorney Docket No.: USC-838-PCT (1726)6575H2N H / H § I 6576N N0!'V / NY Y / 0 / = N / u r^N O X— -O L JH / \\ / / ) z / HI II6577 IZNA M Ty?°~z JH2N H / \z)<^ A H6578 KNN ( V z / l jT j HF2A IH N s7H65790Q JL HN-A J6580OHH2N^ / b\ ^-<x. SA / / ^N\6581... AkTsY ’O Y6582..jY rtA'065916592 A JOL XVA^SH H NH21 H6594 NV / I JUC TAsNN / < Y / X'Y>H2N HPatent Application Attorney Docket No.: USC-838-PCT (1726)6595 o V i XXJ> ZsoHH2NoH H6603 NYVA ' I T nV Vs XV oX HW _ H H6604 Vs\ / VNovNo YoT V sH H6605HoM rNtrYJ H O NX6606H2N A s / X V-S J T XV II — / NH / H HNO N _6607 A A-x Y Y Y nN s Vs XV oHo6609 NA _ < XY fvvNHH2N H / H HNO6613xL i ^ iiH2N s V-NH XV06614 u / \ ^X O XX JXH2N AsNH H1H6616 f^rN^H2N H / 1 H6619A VA Y YY nH2N s v— S XV0o n o X o6621I W "NVVHVPatent Application Attorney Docket No.: USC-838-PCT (1726)Q Hk lH H66220 sftT u F06624 ^fJXnhA -g N / ^•N HHO6625H2N H 00N, y-L'I f jAnh66260HH2N „ H H6628 XL 0XF6631F X - H2N H 00H2N. _ s zz-s6634 Y A- < ji JL JL,nhN-4\ HX~FF F J H H6635 N / N^N'y^r' NH H2 NN-^S 0' N XY— -O L JL^°1 HJ.6636 N A-XXX 1 1 h°H2N As H1 / 6706 N v-XXX JL L >°AsH2N HPatent Application Attorney Docket No.: USC-838-PCT (1726)6637663806639H2NYW ~X XX / - IH2NOH HOz zi6 Vs\ N^N. ^. N 641 X"m' - X®" - / u °XF zVzv cn— cn—ZT ZI jQ H6657X~S N N N NH2N H H0H2NQH6658 Il \ K 9 f| JL T N Q UIh" O / \rs xi?^ / N\^N #~N6663 9 I T Y / >-\ JxAuAtZs"^ S NH, NvH06664H2NYW ~X xy^ N-y6677 yVo. xOC XH2N I H6678H2NX^S / >VSH6681 KX_ / YBYV'NXPatent Application Attorney Docket No.: USC-838-PCT (1726)H2N.. H6670 njYrr'NH,s-r vs kJo6679 ^3 f jL "Nl Cs N. ziHIZ 'H2N |6680z z — ZI—I ZEz z A Z zZ z6681 -A ^z=— - Z> <^^ z z==z y b J I N8 - < N zN C z7V66 1 B I I < N z —I Z <>—H2rA^ V-s ZI L zH66984 4 )=o o z=zTro6699 / H06700 rCYvYYH2N ^ / ~V ' H6701

[0220] Lysyl oxidase family inhibitors disclosed herein may be developed and synthesized according to methods as are generally known in the art. For instance, in one example embodiment, an iterative combinatorial method can be utilized as described in U. S. Patent No. 9,175,357 to Mclnnes. et al., which is incorporated herein by reference. Overall, this combinatorial approach is based upon a known peptide inhibitor and allows both regions of the inhibitor molecule (i.e., the N-cap and the C-cap) to be optimized independently to maximize the affinity and drug-likeness of each component. FIG. 1 provides a general scheme for identification and synthesis of LOX family inhibitors according to this particular embodiment. Peptide inhibitors that can be utilized as the basisPatent Application Attorney Docket No.: USC-838-PCT (1726)for development of the small molecule inhibitors can include any peptide inhibitor capable of selectively inhibiting LOX. For instance, the basis peptide inhibitor can include both native peptides and variants thereof.

[0221] A benefit of the iterative combinatorial formation approach is the modularity in identification of desired moieties for each subsite. The desired fragments can be combinatorically ligated so that essential compound features will not be compromised during the linking process. The individual fragments can be optimized for potency and drug-like properties and then linked to improve pharmacodynamic and pharmacokinetic properties. This lead optimization stage can be informed by cellular assays and detailed mechanistic studies of anti -tumor effects of the inhibitors. The modularity and combinatorial aspects of this method can allow for the moieties to be exchanged and more narrowly target the characteristics of the physicochemical characteristics, as well as minimize metabolic or toxicophore liabilities, with particularly preferred characteristics depending primarily on the application of the inhibitor (e.g., research, in vivo).

[0222] According to one embodiment, following identification and optimization of the end groups, a bridging strategy between the benzamido moiety and an aminobenzoic acid moiety can be selected. For instance, the bridge between the aryl groups can include an amide, sulfonamide, ether, thioether, amine, or carbon-carbon linkages.

[0223] The fragments can be subdivided into chemotypes appropriate for docking into the particular PBD sub-sites and prioritized using pharmacophore features to select fragments with desired functionality. High-throughput docking of virtual libraries into the PBD binding site can be performed during the synthesis process. High-throughput docking (HTD) can then be used in refinement of the fragments with calculations including more accurate scoring functions, and interactions filters to limit fragments to those containing the geometrically appropriate functionality. HTD programs such as, without limitation, Lidaeus, LigandFit, accelrys®, and Glide (Schrodinger®), can incorporate the desired enhancements and can be used to dock virtual fragment libraries into each site. For example, when calculations are parallelized. 100,000 fragments can be screened between 5 and 50 hours depending on the parameterization and number of CPUs employed. Due to inherent inaccuracies with docking, it can be expedient to use different implementations to minimize errors, biases, or incorrect parameters of a single synthesis process. A balance of electrostatic, van der Waals, and H-bonding interactions between each fragment and the binding groove can be used to form the inhibitor having the desired characteristics.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0224] After the development of the inhibitor fragments from identified chemotypes, these can be combined in order to substitute all peptidic determinants and form the non-peptidic small molecule inhibitors. Optimization of the fragments can be facilitated using 3-D structures generated through crystallography and flexible molecular docking (using, e.g., Cdocker, accelrys®, etc.) to predict favorable interactions of modified compounds and improve complementarity.

[0225] In some example embodiments, LOX family inhibitors disclosed herein may be capable of modulating LOX activity. For instance, LOX family inhibitors disclosed herein may be capable of selectively inhibiting or decreasing lysyl oxidase activity compared to the activity of a different protein (e.g., a different protein kinase). In one example embodiment, the selectivity of a LOX inhibitor disclosed herein in inhibiting the activity of LOX over a different protein may be measured by the quotient of the ICso value of the LOX inhibitor in inhibiting the activity of the different protein over the IC50 value of the LOX inhibitor in inhibiting the activity of LOX. It is understood that a compound with a lower IC50 value, as determined under substantially similar conditions, is a more potent inhibitor relative to a compound with a higher IC50 value. Alternatively, the selectivity of LOX inhibitor disclosed herein for LOX over a different protein may be measured by the quotient of the Kd value of an adduct of the inhibitor and the different protein over the Kd value of an adduct of the LOX inhibitor and LOX. In certain embodiments, the selectivity may be at least 2-fold, at least 3-fold, at least 5-fold, at least 10-fold, at least 30-fold, at least 100-fold, at least 300-fold, at least 1,000-fold, at least 3,000-fold, at least 10,000-fold, at least 30,000-fold, or at least 100,000-fold. In some example embodiments, the selectivity may be not more than 100,000-fold, not more than 10,000-fold, not more than 1,000-fold, not more than 100-fold, not more than 10-fold, or not more than 2-fold.

[0226] Advantageously, LOX family inhibitors disclosed herein have been observed as being a competitive and time- and concentration-dependent active site directed irreversible inhibitor. Further, the significant decrease in tumor growth when LOX family inhibitors disclosed herein are combined with chemotherapy in the highly aggressive TNBC PDXs in vivo, encourages further optimization and future clinical testing of our LOX inhibitor. Importantly, LOX family inhibitors disclosed herein have the potential to not only overcome chemoresistance but also to effectively treat other LOX-driven diseases, such as fibrosis.

[0227] In some example embodiments, LOX family inhibitors disclosed herein may be useful for treating a subject in need thereof. The term "treating" as used herein refers toPatent Application Attorney Docket No.: USC-838-PCT (1726)partially or completely alleviating, improving, relieving, inhibiting progression, and / or reducing incidence of one or more symptoms of a disease, disorder, and / or condition, e.g., cancer.

[0228] The term "treating cancer" or "treatment of cancer" may refer to administration of a LOX inhibitor to a subject afflicted with or at risk of a cancerous condition, and may refer to an effect that alleviates the cancerous condition by killing the cancerous cells, but also an effect that result in the inhibition of growth and / or metastasis of the cancer.

[0229] In some embodiments, the cancer may include, but is not limited to, adrenal cortical cancer, advanced cancer, anal cancer, aplastic anemia, bile duct cancer, bladder cancer, bone cancer, bone metastasis, brain tumors, brain cancer, breast cancer, childhood cancer, cancer of unknown primary origin, cervical cancer, colon / rectal cancer, central nervous system (CNS) cancer, endometrial cancer, esophagus cancer, Ewing family of tumors, eye cancer, gallbladder cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, gestational trophoblastic disease, Hodgkin disease, Kaposi sarcoma, renal cell carcinoma, laryngeal and hypopharyngeal cancer, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, chronic myelomonocytic leukemia, liver cancer, non-small cell lung cancer (NSCLC), small cell lung cancer, lung carcinoid tumor, lymphoma of the skin, malignant mesothelioma, multiple myeloma, myelodysplastic syndrome, nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, oral cavity and oropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer, penile cancer, pituitary tumors, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma in adult soft tissue, basal and squamous cell skin cancer, melanoma, small intestine cancer, stomach cancer, testicular cancer, throat cancer, thymus cancer, thyroid cancer, uterine sarcoma, vaginal cancer, vulvar cancer, and / or Waldenstrom macroglobulinemia. For instance, the cancer may include breast cancer, colon cancer, CNS, leukemia, melanoma, prostate, or renal cancer. In one example embodiment, the breast cancer may be triple negative breast cancer (TNBC), estrogen receptor-positive breast cancer, metastatic breast cancer, HER2 positive breast cancer, or a combination thereof. In one example embodiment, the breast cancer may be triple negative breast cancer (TNBC).

[0230] In some example embodiment, the LOXi may be useful in treating a fibrosis-related disease. For instance, the fibrosis-related disease may include, but is not limited to, liver fibrosis, pulmonary fibrosis, renal fibrosis, myocardial fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, radiation-induced fibrosis, ocular fibrosis, Peylony's diseasePatent Application Attorney Docket No.: USC-838-PCT (1726)and scleroderma, or respiratory disorders, abnormal wound healing and repair, postoperative surgery’, cardiac arrest-related fibrosis, excess or abnormal deposition of fibrotic material, all associated with disorders such as Crohn's disease and inflammatory bowel disease, liver, kidney and pancreas fibrosis, diabetes, cerebral hemorrhage with amyloidosis, cardiac hypertrophy, Hutchinson-Gilford Progeria Syndrome, retinopathy, chemoresistance, and / or a kidney disorder including: kidney fibrosis, renal fibrosis, acute kidney injury, chronic kidney disease, diabetic nephropathy, glomerulosclerosis, vesicoureteral reflux, tubulointerstitial renal fibrosis and / or glomerulonephritis.

[0231] The term "subject" refers to any organism to which aspects of the disclosure can be administered, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Subjects to which embodiments of the disclosure can be administered include mammals, such as primates, for example, humans. For veterinary’ applications, a wide variety of subjects are suitable, e.g., livestock such as cattle, sheep, goats, cows, swine, and the like; poultry’ such as chickens, ducks, geese, turkeys, and the like; and domesticated animals, such as pets such as dogs and cats. For diagnostic or research applications, a wide variety of mammals are suitable subjects, including rodents (e.g., mice, rats, hamsters), rabbits, primates, and swine such as inbred pigs and the like. The term "living subject" can refer to a subject noted above or another organism that is alive. The term "living subject" can refer to the entire subject or organism and not just a part excised (e.g., a liver or other organ) from the living subject.

[0232] As used herein, the term "administration" refers to introducing a substance (e.g., a LOX inhibitor, an exogenous antigen, a cytotoxic agent, etc.) into a subject. The administration thereof can be carried out in any convenient manner, including by aerosol inhalation, injection, ingestion, transfusion, implantation, or transplantation. For instance, the cannabinoid compound may be administered orally, subcutaneously, intravenously, or intratumoral. In this regard, "oral" administration can refer to administration into a subject's mouth; "subcutaneous" administration can refer to administration just below the skin; "intravenous" administration can refer to administration into a vein of a subject; and "intratumoral" administration can refer to administration within a tumor.

[0233] Pharmaceutical compositions disclosed herein may be formulated to be compatible with its intended route of administration. As used herein, "routes of administration" may include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), transmucosal. and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include thePatent Application Attorney Docket No.: USC-838-PCT (1726)following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

[0234] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EM™ (BASF, Parsippany, N. J.) or phosphate buffered saline (PBS). The composition can be sterile and should be fluid to the extent that easy syringability exists. It can be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, a pharmaceutically acceptable polyol like glycerol, propylene glycol, liquid polyethylene glycol, and suitable mixtures thereof. Prolonged absorption of the injectable compositions may be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

[0235] Oral compositions may include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier may be applied orally.

[0236] Pharmaceutically compatible binding agents, and / or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or com starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such asPatent Application Attorney Docket No.: USC-838-PCT (1726)sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.

[0237] Compositions for parenteral delivery, e.g., via injection, can include pharmaceutically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions, or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (e.g., com oil) and injectable organic esters such as ethyl oleate. In addition, the composition can contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like that can enhance the effectiveness of the phenolic compound. Proper fluidity may be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants. These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents.

[0238] In one embodiment, a therapeutically effective amount of the LOXi may be administered to the subject. The term "therapeutically effective amount" refers to those amounts that, when administered to a subject in view of the nature and severity of that subject's disease or condition, will have a desired therapeutic effect, e.g., an amount which will cure, prevent, inhibit, or at least partially arrest or partially prevent a target disease or condition. A therapeutically effective dose further can refer to that amount of the therapeutic agent sufficient to result in amelioration of symptoms, e.g., treatment, healing, prevention or amelioration of the relevant medical condition, or an increase in rate of treatment, healing, prevention or amelioration of such conditions. When applied to an individual active ingredient administered alone, a therapeutically effective dose can refer to that ingredient alone. When applied to a combination, a therapeutically effective dose can refer to combined amounts of the active ingredients that result in the therapeutic effect, whether administered in combination, serially or simultaneously.

[0239] A therapeutically effective dose can depend upon a number of factors known to those of ordinary' skill in the art. The dosage can vary' depending upon know n factors such as the pharmacodynamic characteristics of the active ingredient and its mode and route of administration; time of administration of active ingredient; identity, size, condition, age, sex, health and weight of the subject or sample being treated; nature and extent ofPatent Application Attorney Docket No.: USC-838-PCT (1726)symptoms; kind of concurrent treatment, frequency of treatment and the effect desired; and rate of excretion. These amounts can be readily determined by the skilled artisan.

[0240] In one example embodiment, the therapeutically effective amount is at least about 0.1 mg / kg body weight, at least about 0.25 mg / kg body weight, at least about 0.5 mg / kg body weight, at least about 0.75 mg / kg body weight, at least about 1 mg / kg body weight, at least about 2 mg / kg body weight, at least about 3 mg / kg body weight, at least about 4 mg / kg body weight, at least about 5 mg / kg body weight, at least about 6 mg / kg body weight, at least about 7 mg / kg body weight, at least about 8 mg / kg body weight, at least about 9 mg / kg body weight, at least about 10 mg / kg body weight, at least about 15 mg / kg body weight, at least about 20 mg / kg body weight, at least about 25 mg / kg body¬ weight. at least about 30 mg / kg body weight, at least about 40 mg / kg body weight, at least about 50 mg / kg body weight, at least about 75 mg / kg body weight, at least about 100 mg / kg body weight.

[0241] In some embodiments, for instance, the LOXi may be administered to a subject at a dosage of about 20 mg / kg body weight to about 100 mg / kg body weight, such as from about 25 mg / kg body weight to about 75 mg / kg body weight, such as from about 30 mg / kg body weight to about 60 mg / kg body weight, or any7range therebetween.

[0242] Pharmaceutical compositions as described herein can be administered to the subject one time (e.g., as a single injection or deposition). Alternatively, administration can be once or twice daily to a subject in need thereof for a period of from about 2 days to about 35 days, such as from about 7 days to about 28 days, such as from about 10 to about 21 days, or any range therebetween. It can also be administered once or twice daily to a subject for a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 times per year, or a combination thereof.

[0243] In some example embodiments, LOXi disclosed herein may be utilized in methods for re-sensitizing a cancer cell. A cancer cell is sensitive to an anti-cancer compound when the cancer cell is killed or the grow th and / or spread of the cancer cell is inhibited by contacting the cancer cell with the anti-cancer compound. Cancer cells may also be resistant to an anti-cancer compound, wherein contacting the cancer cell with the anti-cancer compound does not kill or inhibit the growth and / or spread of the cancer cell. Resistance may be inherent to the cancer cell, wherein the anti-cancer reagent never kills or inhibits the growth and / or spread of the cancer cell. Resistance may also be acquired, wherein the cancer cell is initially sensitive to the anti-cancer compound, but over time the cancer cell becomes resistant. Sensitivity of a cancer cell to an anti-cancer compound mayPatent Application Attorney Docket No.: USC-838-PCT (1726)be determined based on any method known in the art including, cell mass, proliferation, survival, metastasis, and / or expression of cell surface markers.

[0244] In some example embodiments, the cancer cell is sensitive to the anti-cancer compound when the mass of the cell contacted with the anti-cancer compound decreases compared to a control cell. A control cell may be a normal (e.g., non-cancerous cell). A control cell may be another cell derived from the same tissue in the subject that does not include cancer cells, a cell that was previously cancerous but is no longer cancerous, or a cell from another subject that is derived from the same tissue type as the cancer cell. In some example embodiments, when the cancer cell is resistant to the anti-cancer compound, the mass of the cell contacted with the anti-cancer compound increases or stays the same compared to the control cell. In order to compare the mass of single cells (e.g., primary cancer cells versus control cells), the mass of the single cells being compared is normalized.

[0245] In one example embodiment, the cancer or cells of the cancer may be resistant to primary cancer therapeutic treatment prior to administering the LOX inhibitor. In one example embodiment, the cancer or cells of the cancer may be resistant to doxorubicin prior to administering the LOX inhibitor. In another example embodiment, the cancer or cells of the cancer may be resistant to docetaxel prior to administering the LOX inhibitor. Thus, the LOX inhibitor may be administered to a subject in an amount effective to resensitize the cancer cells to primary cancer therapeutic treatment.

[0246] In another example embodiment, the cancer or cells of the cancer may not be resistant to primary cancer therapeutic treatment, and the LOX inhibitor may be administered in an amount effective to inhibit, slow, or prevent cancer cell resistance to primary’ cancer therapeutic treatment.

[0247] If desired, in some example embodiments, in methods for re-sensitizing a cancer cell, the LOX family inhibitors disclosed herein may be administered to a subject in combination with one or more chemotherapeutic agents. In one example embodiment, the one or more chemotherapeutic agents may include, but are not limited to, anthracycline, doxorubicin, cisplatin, or paclitaxel. In another example embodiment, the one or more chemotherapeutic agents may be an anti-PD-1 antibody including, but is not limited to, pembrolizumab, nivolumab, dostarlimab, camrelizumab, spartalizumab, sintilimab, or cemiplimab. In another example embodiment, the one or more chemotherapeutic agents may be an anti-PD-Ll antibody including, but is not limited to, atezolizumab, avelumab, cosibelimab, envafolimab, or durvalumab.Patent Application Attorney Docket No.: USC-838-PCT (1726)Table B: AbbreviationsCOL6A2 Collagen Type VI Alpha 2 ChainCOL6A3 Collagen Type VI Alpha 3 ChainPOSTN PeriostinCOL5A1 Collagen Type V Alpha 1 ChainCOL5A2 Collagen Type V Alpha 2 ChainTSBH2- Thrombospondin 2-1TSBH1TNC Tenascin CTGFB1 Transforming Growth Factor Beta 1VIM VimentinBGN BiglycanFN1 Fibronectin 1COL1A2 Collagen Type I Alpha 2 ChainCOL6A1 Collagen Type VI Alpha 1 ChainLUM LumicanCOL1A1 Collagen Type I Alpha 1 ChainFGB Fibrinogen Beta ChainH1ST1H2BA H2B Clustered Histone 1ANXA2 Annexin A2PPIB Peptidylprolyl Isomerase BDHX9 DExH-Box Helicase 9COL3A1 Collagen Type III Alpha 1 ChainLMNA Lamin A / CLRP1 LDL Receptor Related Protein 1LGALS3 Galectin 3PEBP1 Phosphatidylethanolamine Binding Protein 1 P4HB Prolyl 4-Hydroxylase Subunit BetaGSN GelsolinDST DystoninVDAC1 Voltage Dependent Anion Channel 1SERPINH1 Serpin Family H Member 1TLN1 Talin 1MYH10 Myosin Heavy Chain 10ENO1 Enolase 1Protein Disulfide Isomerase Family A MemberPDIA66DCC DCC Netrin 1 ReceptorPLEC Plectin

[0248] The preceding description is exemplary in nature and is not intended to limit the scope, applicability or configuration of the disclosure in any way. Various changes to the described embodiments may be made in the function and arrangement of the elements described herein without departing from the scope of the disclosure.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0249] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention is related.

[0250] As used in this application and in the claims, the singular forms "a", "an", and "the" include the plural forms unless the context clearly dictates otherwise. Additionally, the term "includes" means "comprises". The methods and compositions of the present disclosure, including components thereof, can comprise, consist of, or consist essentially of the essential elements and limitations of the embodiments described herein, as well as any additional or optional ingredients, components or limitations described herein or otherwise useful in an agent disclosed herein.

[0251] Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as molecular weight, percentages, and so forth, as used in the specification or claims are to be understood as being modified by the term "about". Accordingly, unless otherwise indicated, implicitly or explicitly, the numerical parameters set forth are approximations that may depend on the desired properties sought and / or limits of detection under standard test conditions / methods. When directly and explicitly distinguishing embodiments from discussed prior art, the embodiment numbers are not approximate unless the word "about" is recited.

[0252] As used herein, "optional" or "optionally" means that the subsequently- described material, event or circumstance may or may not be present or occur, and that the description includes instances where the material, event or circumstance is present or occurs and instances in which it does not. As used herein, "w / w%" and "wt%" mean by weight as relative to another component or a percentage of the total weight in the composition.

[0253] The term "about" is intended to mean approximately, in the region of, roughly, or around. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. Unless otherwise indicated, it should be understood that the numerical parameters set forth in the following specification and attached claims are approximations. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, numerical parameters should be read in light of the number of reported significant digits and the application of ordinary rounding techniques.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0254] The phrase "effective amount" means an amount of a compound that promotes, improves, stimulates, or encourages a response to the particular condition or disorder or the particular symptom of the condition or disorder.

[0255] As used herein, an “alkyl” group may refer to a straight or branched chain hydrocarbon, having a certain number of carbon atoms (e.g., C1-12 carbon atoms). For instance, an alkyl group may include, but is not limited to, straight and branched chain alkyl groups having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. Alternatively, an alkyl group may include, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, or a combination thereof.

[0256] As used herein, an “aryl” group may refer to, by itself or as part of another substituent, may include, but is not limited to, a monocyclic, bicyclic or polycyclic polyunsaturated aromatic hydrocarbon radical containing 6 to 14 ring carbon atoms, which may be a single ring or multiple rings (up to three rings) which are fused together or linked covalently.

[0257] As used herein, an “alkenyl” group may refer to a linear monovalent hydrocarbon radical or a branched monovalent hydrocarbon radical having the number of carbon atoms indicated in the prefix and containing at least one double bond. Additionally, as used herein, “alkynyl” may refer to a linear monovalent hydrocarbon radical or a branched monovalent hydrocarbon radical containing at least one triple bond and having the number of carbon atoms indicated in the prefix.

[0258] As used herein, an “alkoxy” group may refer to an -O-alkyl group, where alkyl is as defined herein. Similarly, an “halogen substituted alkoxy” may refer to an alkoxy in which the alkyl group is substituted with one or more halogen atoms. Additionally, “thioalkoxy” group may refer to an -O-alkylthio group, where the alkylthio group may include, but is not limited to, thiomethoxy, thioethoxy, and the like.

[0259] As used herein, a “halogen” group may refer to an element found in Group 17 (formerly Group VII A) of the periodic table. For instance, a halogen may include, but is not limited to, fluorine (F), chlorine (Cl), bromine (Br), iodine (I), astatine (At), and the like.

[0260] “Cycloalkyl” refers to a partially or fully saturated, monocyclic, or polycyclic carbocyclic ring, which may include fused (when fused with an aryl or a heteroaryl ring, the cycloalkyl is bonded through a non-aromatic ring atom), spiro, or bridged ring systems. In some embodiments, the cycloalkyl is fully saturated. Monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.Patent Application Attorney Docket No.: USC-838-PCT (1726)Polycyclic cycloalkyls include, for example, adamantyl, norbornyl, decalinyl, bicyclo[3.3.0]octane. bicyclo[4.3.0]nonane, cis-decalin, trans-decalin, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, bicyclo[3.2.2]nonane, and bicyclo [3.3.2] decane, and 7,7-dimethyl-bicyclo[2.2.1]heptanyl. Partially saturated cycloalkyls include, for example cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl.

[0261] ''Heterocycloalkyl” refers to a 4- to 10-membered partially or fully saturated ring radical comprising 2 to 23 carbon atoms and from one to 8 heteroatoms selected from the group consisting of nitrogen, oxygen, phosphorous, silicon, and sulfur. In some embodiments, the heterocycloalkyl is fully saturated. In some embodiments, the heterocycloalkyl comprises one to three heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. Unless stated otherwise specifically in the specification, the heterocycloalkyl radical may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused (when fused with an aryl or a heteroaryl ring, the heterocycloalkyl is bonded through a non-aromatic ring atom), spiro, or bridged ring systems; and the nitrogen, carbon, or sulfur atoms in the heterocycloalkyl radical may be optionally oxidized; the nitrogen atom may be optionally quaternized. Examples of such heterocycloalkyl radicals include, but are not limited to, aziridinyl, azetidinyl, oxetanyl, dioxolanyl, thienyl[l,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl. octahydroisoindolyl. 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl. thiamorpholinyl, 1-oxo-thiomorpholinyl, 1,1-dioxo-thiomorpholinyl, 1,3-dihydroisobenzofuran-1-yl, 3-oxo-1,3-dihydroisobenzofuran-1-yl, methyl-2-oxo-l,3-dioxol-4-yl, and 2-oxo-l,3-dioxol-4-yl.

[0262] ’‘Heteroaryl” refers to a 5- to 10-membered ring system radical comprising one to thirteen carbon atoms, one to six heteroatoms selected from the group consisting of nitrogen, oxygen, phosphorous, and sulfur, and at least one aromatic ring. The heteroaryl radical may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused (when fused with a cycloalkyl or heterocycloalkyl ring, the heteroaryl is bonded through an aromatic ring atom) or bridged ring systems; and the nitrogen, carbon, or sulfur atoms in the heteroaryl radical may be optionally oxidized; the nitrogen atom may be optionally quaternized. In some embodiments, the heteroaryl is a 5- to 6-membered heteroaryl. In some embodiments, the heteroaryl is a 6-membered heteroaryl. In somePatent Application Attorney Docket No.: USC-838-PCT (1726)embodiments, the heteroaryl is a 5-membered heteroaryl. Examples include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][l,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[l,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1 -oxi dopy razinyl, 1-oxidopyridazinyl, 1-phenyl-lH-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl. phthalazinyl. pteridinyl, purinyl, pyrrolyl. pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl).

[0263] Here and throughout the specification and claims, range limitations are combined and interchanged, such ranges are identified and include all the sub-ranges contained therein unless context or language indicates otherwise. For example, all ranges disclosed herein are inclusive of the endpoints, and the endpoints are independently combinable with each other.

[0264] This written description uses examples to disclose the present disclosure, including the best mode, and also to enable any person skilled in the art to practice the disclosure, including making and using any devices or systems and performing any incorporated methods. The patentable scope of the disclosure is defined by the claims, and may include other examples that occur to those skilled in the art. Such other examples are intended to be within the scope of the claims if they include structural elements that do not differ from the literal language of the claims, or if they include equivalent structural elements with insubstantial differences from the literal languages of the claims.

[0265] According to one embodiment, disclosed herein is a compound of FormulaFormula (A),Patent Application Attorney Docket No.: USC-838-PCT (1726)or a pharmaceutically acceptable salt thereof, wherein:

[0267] X is CH or N;

[0268] R1is C1-8alkyl or C1-8haloalkyl;

[0269] R2and R3are independently hydrogen, Ci-8 alkyl, Ci-8 alkylamino, -CH2-phenyl, or -C(O)Ra; wherein each Rais independently C1-8 alkyl or phenyl;

[0270] R4, R5, R6, and R7are each independently hydrogen, halogen, C1-8 alkyl, C1-8 alkylamino, C1-8 alkoxy. -ORb, -N(Rb)2. -C(O)ORb. -C(O)RC. -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc,

[0271] or R4and R5or R6and R7combine with the intervening atoms to form a phenyl, 5 to 6-membered heteroaryl, or 5 to 6-membered heterocycloalkyl which is saturated or partially unsaturated, wherein the phenyl, heteroaryl and heterocycloalkyl are optionally substituted with one or more R8;

[0272] each R8is independently halogen, C1-8 alkyl, -ORb, oxo (=0), -N(Rb)2, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc;

[0273] each Rbis independently hydrogen, C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd;

[0274] or two Rbcombine with the nitrogen atom to which they are attached to forms a 4 to 8-membered heterocycloalkyl which is optionally substituted with one or more

[0275] each Rcis independently C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; and

[0276] each Rdis independently halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, wherein the phenyl or heteroaryl is optionally substituted with one or more halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), -CH3, -C(O)OH, -C(O)OCH3, or -NHC(O)CH3.

[0277] In one example embodiment, when ring A of Formula (a) is, then at least one of R4, R5, R6, or R7is not hydrogen.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0278] According to one example embodiment, the compound is of Formula (1-4):or a pharmaceutically acceptable salt thereof, wherein:X is CH orN;R1is -CH3, -CH2CH3, or -CF3;R2and R3are independently hydrogen, C1-8 alkyl, or -C(O)Ra; wherein each Rais independently C1-8 alkyl or phenyl;R4, R5, R6, and R7are each independently hydrogen, halogen, C1-8 alkyl, C1-8 alkylamino, Ci-8 alkoxy, -ORb, -N(Rb)2, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc,or R4and R5or R6and R7combine with the intervening atoms to form a phenyl, 5 to 6-membered heteroaryl, or 5 to 6-membered heterocycloalkyl which is saturated or partially unsaturated, wherein the phenyl, heteroaryl, and heterocycloalkyl is optionally substituted with one or more R8, each R8is independently halogen, C1-8 alkyl, -ORb, oxo (=0), -N(Rb)2, -C(O)ORb, - C(O)RC. -C(0)N(Rb)2, or -NRbC(O)Rc;each Rbis independently hydrogen, C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; or two Rbcombine with the nitrogen atom to which they are attached to forms a 4 to 8-membered heterocycloalkyl which is optionally substituted with one or more Rd;each Rcis independently C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; andeach Rdis independently halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, wherein the phenyl or heteroaryl is optionally substituted with one or more halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), -CH3, -C(O)OH, -C(O)OCH3, or -NHC(O)CH3;Patent Application Attorney Docket No.: USC-838-PCT (1726)R5RI X ^R6V X Rwherein when ring Ais '7, then at least one of R4, R5, R6, or R7is not hydrogen.

[0279] In one example embodiment, X of Formula (1-4) may be N.

[0280] In one example embodiment, X of Formula (1-4) may be CH.

[0281] In one example embodiment, wherein R5of Formula (1-4) may be hydrogen.

[0282] In one example embodiment, R4of Formula (1-4) may be hydrogen, - C(O)ORb, -C(O)RC, or -NRbC(O)Rc.

[0283] In one example embodiment. R6and R7of Formula (1-4) may be each independently hydrogen, halogen, Ci-s alkylamino, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc; wherein one of R6or R7is not hydrogen.

[0284] In one example embodiment, X of Formula (1-4) may be N;R4of Formula (1-4) may be hydrogen; R5of Formula (1-4) may be hydrogen; R6of Formula (1-4) may be hydrogen, halogen, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc; and R7of Formula (1-4) may be halogen, Ci-s alkylamino, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2Rc, -S(O)2N(Rb)2, or -NRbS(O)2Rc.

[0285] In one example embodiment. R6of Formula (1-4) may be hydrogen or halogen.

[0286] In one example embodiment, R7of Formula (1-4) may be -C(O)OH, -NHC(O)CH3, -NHC(O)CH2CH2OCH3, -C(O)NH-(CH2)4-phenyl, -C(O)NH-phenyl, -C(O)NH-(2,4-difluoro-phenyl), or -C(O)NH-phenyl-4-acetamide.

[0287] In one example embodiment, X of Formula (1-4) may be CH; R4of Formula (1-4) may be hydrogen; R5of Formula (1-4) may be hydrogen; R6of Formula (1-4) may be hydrogen, halogen, Ci-s alkylamino, -C(O)ORb, -C(O)RC, -NRbC(O)Rc, -S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc; and R7of Formula (1-4) may be hydrogen, halogen, Ci-8 alkylamino, -C(O)ORb, -C(O)RC, -NRbC(O)Rc, -S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc: wherein one of R6or R7is not hydrogen.

[0288] In one example embodiment, R6of Formula (1-4) may be hydrogen or halogen; and R7of Formula (1-4) may be -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0289] In one example embodiment, R7of Formula (1-4) may be -CH2NH2, -C(O)OH. -C(O)NHCH3, -C(O)N(CH3)2. -NHC(O)CH3, -NHC(O)CH2CH2OCH3. -C(O)NH-(CH2)4-phenyl, -C(O)NH-phenyL -C(O)NH-(2,4-difluoro-phenyl), -C(O)NH-phenyl-4-acetamide, -NHC(O)-CH2-(4-fluoro-phenyl), -NHC(O)-CH2-(4-cholo-phenyl), -NHC(O)-(4-methoxy-phenyl), -NHC(O)-3-pyridynyl, or -NHSO2CH3.

[0290] In one example embodiment, R4and R5of Formula (1-4) may be combined together with the intervening atoms to form a 5 to 6-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S, or a 5 to 6-membered heterocycloalkyl with is saturated or partially unsaturated, having 1 or 2 heteroatoms selected from N and O, wherein the heteroaryl or heterocycloalkyl is optionally substituted with one or more R8.

[0291] In one example embodiment. R4and R5of Formula (1-4) may be combined with the intervening atoms to form a pyrrole, imidazole, triazole, thiazole, oxazole, pyridine, furan, pyrrolidine, pyrrolidinone, imidazolidinone, tetrahydropyran, or piperidinone.

[0292] In one example embodiment, R4and R5of Formula (1-4) may be combined with the intervening atoms to form a pyrrolidine, pyrrolidinone, or imidazolidinone.

[0293] In one example embodiment, R6and R7of Formula (1-4) may be combined together with the intervening atoms to form a 5 to 6-membered heteroaryl having 1, 2, or 3 heteroatoms selected from N, O, and S, or a 5 to 6-membered heterocycloalkyl with is saturated or partially unsaturated, having 1 or 2 heteroatoms selected from N and O, wherein the heteroaryl or heterocycloalkyl is optionally substituted with one or more R8.

[0294] In one example embodiment, R5and R6of Formula (1-4) may be combined together with the intervening atoms to form a pyrrole, imidazole, triazole, thiazole, oxazole, pyridine, furan, pyrrolidine, pyrrolidinone, imidazolidinone, tetrahydropyran, or piperidinone.

[0295] In one example embodiment, R4and R5of Formula (1-4) may be combined together with the intervening atoms to form a pyrrolidine, pyrrolidinone, or imidazolidinone.

[0296] According to one example embodiment, the compound is of Formula (1-3):or a pharmaceutically acceptable salt thereof, wherein:Patent Application Attorney Docket No.: USC-838-PCT (1726)R1is -CH3, -CH2CH3, or -CF3;R2and R3are independently hydrogen, C1-8 alkyl, or -C(O)Ra; wherein each Rais independently C1-8 alkyl or phenyl;R4is hydrogen, C1-8 alkyl, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc;R5is hydrogen, C1-8 alkyl, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc;each Rbis independently hydrogen, C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd;or two Rbcombine together with the nitrogen atom to which they are attached to forms a 4 to 8-membered heterocycloalkyl which is optionally substituted with one or more Rd; each Rcis independently C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; andeach Rdis independently halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH ), Ci-s alkyl, phenyl, or 5 to 6-membered heteroaryl, wherein the phenyl or heteroaryl is optionally substituted with one or more halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), -CH3, -C(O)OH. -C(O)OCH3, or -NHC(O)CH3.

[0297] In one example embodiment, R4of Formula (1-3) may be hydrogen, -NHC(O)CHs, or -C(O)OCHsCH; and R' of Formula (1-3) may be hydrogen, -NHC(O)CH3, -NHC(O)-Ph-NHC(O)CH3, -NHC(O)-Ph-2,4-difluoro, -C(O)OH, - C(O)NHPh, or -C(O)NH-Ph-NHC(O)CH3.

[0298] In one example embodiment. R4of Formula (1-3) may be hydrogen.

[0299] In one example embodiment, R5of Formula (1-3) may be hydrogen.

[0300] In one example embodiment, R1of Formula (1-3) may be -CH3.

[0301] In one example embodiment, R2of Formula (1-3) may be hydrogen, -CH3, - CH2CH3, or -C(O)CH3.

[0302] In one example embodiment, R2of Formula (1-3) may be hydrogen.

[0303] In one example embodiment, R3of Formula (1-3) may' be hydrogen or -CHs.

[0304] According to one example embodiment, a compound may be selected from:Patent Application Attorney Docket No.: USC-838-PCT (1726)6426 A?A- / H2N H H I / H HN'A _N. / 6487H2N s V-S T Y 0 n6488N^L _ JATYH2N A-s' HH6489H2N Ab<; H L.oz1H6505 N'L / 'i ATNAZ-N HH / H HO NA N^ / N N N^ / 6506AAK T V r / H HNA N^N^\. N. / 6512A AA r r YH2N S V-S06519-A ° jYd JCOy' N jH6519-B o rUl -00ANHJ k<y ~6522NA-Y3 lAH2N H6523NJ A.— r fV | y YOAs' ' N^N°H2N H6524ik i-k ro,H2N HPatent Application Attorney Docket No.: USC-838-PCT (1726)6528H2N06 2 / HF5 9HH2N 4<x H,n"rc,1 A A6530 N-V / l JLJ HH2N As H06531H2N ik A H1 HNA / I I M / ) 65322As \ / H N HSy> NH2N6533A 'H2N As^ ^ H038H2NysZN>, NH 652N^~VJ o6539H2N xHx. HxrV06554 MA / '"Sf^ir NH^N AA A JL IJH2N As HPatent Application Attorney Docket No.: USC-838-PCT (1726)H6555N A-A A L l| HH2N As Ho6556 N1_Z IH2N H6558 / 7-QN4 _A \ T nH2N H0 A'N6559H2N H06560 NA__ / AH2N |j?6561H2N As' Hh|| i j ofHA? f6562 II / 'AsNN N A< A H2N HhH I6563 ifL / i n xvAH2AS Nn HNH NH26568H2N As' ^ Hoh6569H2N As^ ^ HHNr^Patent Application Attorney Docket No.: USC-838-PCT (1726)6570, Py z—As' ' _ / T6571ZIHNO A HL zNA--O6572 A - - -SNH2N hHN^O< N zI6573H2N As' ' H6574NV-ZA 1 1 JH2N HJ6574-B NA^-A I I T IHAs^N^N^^Af^H2N H IIJ / -q6574-C NA— A I I T I H XSA^NA AZNXH2N I IIs-^\ S^NH2>x / x ii0 HN N V-N6574-DAHA JI I IH6574-E H2N _ T nys 0NT~V7Patent Application Attorney Docket No.: USC-838-PCT (1726)6575H2N H / H § I 6576N N0!'V / NY Y / 0 / = N / u r^N O X— -O L JH / \\ / / ) z / HI II6577 IZNA M Ty?°~z JH2N H / \z)<^ A H6578 KNN ( V z / l jT j HF2A IH N s7H65790Q JL HN-A J6580OHH2N^ / b\ ^-<x. SA / / ^N\6581... AkTsY ’O Y6582..jY rtA'065916592 A JOL XVA^SH H NH21 H6594 NV / I JUC TAsNN / < Y / X'Y>H2N HPatent Application Attorney Docket No.: USC-838-PCT (1726)6595 o V i XXJ> ZsoHH2NoH H6603 NYVA ' I T nV Vs XV oX HW _ H H6604 Vs\ / VNovNo YoT V sH H6605HoM rNtrYJ H O NX6606H2N A s / X V-S J T XV II — / NH / H HNO N _6607 A A-x Y Y Y nN s Vs XV oHo6609 NA _ < XY fvvNHH2N H / H HNO6613xL i ^ iiH2N s V-NH XV06614 u / \ ^X O XX JXH2N AsNH H1H6616 f^rN^H2N H / 1 H6619A VA Y YY nH2N s v— S XV0o n o X o6621I W "NVVHVPatent Application Attorney Docket No.: USC-838-PCT (1726)Q Hk lH H66220 sftT u F06624 ^fJXnhA -g N / ^•N HHO6625H2N H 00N, y-L'I f jAnh66260HH2N „ H H6628 XL 0XF6631F X - H2N H 00H2N. _ s zz-s6634 Y A- < ji JL JL,nhN-4\ HX~FF F J H H6635 N / N^N'y^r' NH H2 NN-^S 0' N XY— -O L JL^°1 HJ.6636 N A-XXX 1 1 h°H2N As H1 / 6706 N v-XXX JL L >°AsH2N HPatent Application Attorney Docket No.: USC-838-PCT (1726)6637663806639H2NYW ~X XX / - IH2NOH HOz zi6 Vs\ N^N. ^. N 641 X"m' - X®" - / u °XF zVzv cn— cn—ZT ZI jQ H6657X~S N N N NH2N H H0H2NQH6658 Il \ K 9 f| JL T N Q UIh" O / \rs xi?^ / N\^N #~N6663 9 I T Y / >-\ JxAuAtZs"^ S NH, NvH06664H2NYW ~X xy^ N-y6677 yVo. xOC XH2N I H6678H2NX^S / >VSH6681 KX_ / YBYV'NXPatent Application Attorney Docket No.: USC-838-PCT (1726)H2N.. H6670 njYrr'NH,s-r vs kJo6679 N. ^3 f jL "Nl CsZziHI 'H2N |6680z — ZIz —I ZEz Z z A Z z L z6681 -A ^z=— - Z> <^^ z z==y b< N zN C z z J I N6681 -B J W7VV z — VI ZE I < N Z <>— Y VH2N^S^ V-S ZI L z M^CIH6698wz z4 4 )=o o z=z IZ T\ro6699 / H06700HrLYYYY 2N^ / ~V' H6701

[0305] In one example embodiment, the compound, or pharmaceutically acceptable salt thereof, may be a lysyl oxidase (LOX) inhibitor.

[0306] In one example embodiment, the compound, or a pharmaceutically acceptable salt thereof, may inhibit cell-based lysyl oxidase activity and / or recombinant protein based lysyl oxidase activity assays.

[0307] In one example embodiment, the compound, or pharmaceutically acceptable salt thereof, may be selective for LOX.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0308] According to one example embodiment, disclosed herein is a pharmaceutically composition comprising a compound disclosed in an example embodiment herein, or a pharmaceutically acceptable salt thereof.

[0309] According to one example embodiment, disclosed herein is a method of treating a disease of condition associated with the modulation or inhibition of lysyl oxidase (LOX), the method comprising administering to a subject in need thereof, a therapeutically effective amount of a compound of disclosed in an example embodiment herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed in an example embodiment herein.

[0310] In one example embodiment, the therapeutically effective amount of the compound or its pharmaceutically acceptable salt may be administered to treat a neurodegenerative disease, an angiogenesis-related disease, Alzheimer's disease, fibrosis including: liver fibrosis, pulmonary fibrosis, renal fibrosis, myocardial fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, radiation-induced fibrosis, ocular fibrosis, Peylony's disease and scleroderma, or respiratory disorders, abnormal wound healing and repair, postoperative surgery, cardiac arrest-related fibrosis, excess or abnormal deposition of fibrotic material, all associated with disorders such as Crohn's disease and inflammatory bowel disease, liver, kidney and pancreas fibrosis, diabetes, cerebral hemorrhage with amyloidosis, cardiac hypertrophy, Hutchinson-Gilford Progeria Syndrome, retinopathy, chemoresistance, and / or a kidney disorder including: kidney fibrosis, renal fibrosis, acute kidney injury, chronic kidney disease, diabetic nephropathy, glomerulosclerosis, vesicoureteral reflux, hypertrophic scarring, keloids, diabetic skin, tubulointerstitial renal fibrosis and / or glomerulonephritis.

[0311] In one example embodiment, the cancer may be selected from the group comprising lung cancer; breast cancer; colorectal cancer; anal cancer; pancreatic cancer; prostate cancer; ovarian carcinoma; liver and bile duct carcinoma; esophageal carcinoma; non-Hodgkin's lymphoma; bladder carcinoma; carcinoma of the uterus; glioma, glioblastoma, medullablastoma, and other tumors of the brain; myelofibrosis, kidney cancer; cancer of the head and neck; cancer of the stomach; multiple myeloma; testicular cancer; germ cell tumor; neuroendocrine tumor; cervical cancer; oral cancer, carcinoids of the gastrointestinal tract, breast, and other organs; signet ring cell carcinoma; mesenchymal tumors including sarcomas, fibrosarcomas, haemangioma, angiomatosis, haemangiopericytoma. pseudoangiomatous stromal hyperplasia, myofibroblastoma, fibromatosis, inflammatory myofibroblastic tumour, lipoma, angiolipoma, granular cellPatent Application Attorney Docket No.: USC-838-PCT (1726)tumour, neurofibroma, schwannoma, angiosarcoma, liposarcoma, rhabdomyosarcoma, osteosarcoma, leiomyoma and / or a leiomysarcoma.

[0312] In one example embodiment, the breast cancer may be triple negative breast cancer (TNBC), estrogen receptor-positive breast cancer, metastatic breast cancer, HER2 positive breast cancer, or a combination thereof.

[0313] In one example embodiment, the therapeutically effective amount of the compound or its pharmaceutically acceptable salt may be administered to reduce cardiotoxicity side effects of chemotherapy and / or immunotherapy.

[0314] In one example embodiment, the therapeutically effective amount of the compound or its pharmaceutically acceptable salt may be administered as a single agent or in combination with chemotherapy, immunotherapy and / or radiotherapy in both adjuvant and neo-adjuvant settings.

[0315] In one example embodiment, the therapeutically effective amount of the compound or its pharmaceutically acceptable salt may be administered with a second therapeutic agent selected from an anti-cancer agent, an anti-inflammatory agent, an antihypertensive agent, an anti -fibrotic agent, an anti-angiogenic agent, and / or an immunosuppressive agent.

[0316] In one example embodiment, the subject may have failed at least one anticancer therapy.

[0317] Furthermore, certain aspects of the present disclosure may be better understood according to the following examples, which are intended to be non-limiting and exemplary in nature. Moreover, it will be understood that the compositions described in the examples may be substantially free of any substance not expressly described.

[0318] The present disclosure may be better understood with reference to the following examples.EXAMPLESMaterials and MethodsCell Culture and reagents

[0319] Human TNBC cell lines, MDA-MB-231, HCC1937, Hs-578-T, and HCC 1143, and normal cells, MCF-12A, HUVEC and HFF-1 were obtained from ATCC and cultured in Dulbecco Modified Eagle Medium (Gibco, USA) supplemented with 50 U / ml penicillin / streptomycin, 1% non-essential amino acids and 10% fetal bovine serum (Gibco, USA) except HUVEC cells. HUVEC cells were cultured in F-12K medium (ATCC)Patent Application Attorney Docket No.: USC-838-PCT (1726)supplemented with 50 U / ml penicillin / streptomycin, 1% non-essential amino acids, 10% fetal bovine serum. 0.1 mg / mL heparin and 0.05 mg / mL ECGS. MCF-12A cells were also supplemented with 20 ng / ml epidermal growth factor (EGF) and 500 ng / ml hydrocortisone. Cells were routinely tested for mycoplasma contamination using MycoAlert detection kit (Lonza) and were authenticated by STR sequencing. The cumulative culture length of cells between thawing and use in this study was less than 20 passages.TNBC PDX xenograft mice tumor model

[0320] 2-3 mm3of PDX tumor pieces from TNBC PDX model TM01278 were transplanted to the flank region of the NSG mouse. When the tumors reached 100 mm’, mice were randomly distributed to 4 groups and treated with vehicle (PEG400 (50%)-Tween80 (l%)-acetate buffer (49%)), doxorubicin (2 mg / kg once a week, I. V.), LXG6403 (50 mg / kg bi-daily, oral gavage) or the combination of doxorubicin with LXG6403. Tumor volumes were measured using a caliper twice a week, and body weights were also recorded. After 24 days of treatment, mice were sacrificed.Western blotting

[0321] Briefly, RIPA buffer was used to isolate total protein lysate in the presence of protease and phosphatase inhibitor cocktails, and protein concentrations were measured using the BCA Protein Assay Reagent Kit (Thermo Fisher Scientific. MA, USA). For extracting proteins from collagen-embedded cells, cells were treated with 1.5 mg / ml of collagenase solution for 5 minutes at 37 °C. Equal amounts of protein were separated using 8-10% SDS-PAGE gel. Separated proteins were transferred to PVDF membranes (BioRad, CA. USA) using a Trans-Blot turbo transfer system (Bio-Rad. CA, USA) and incubated with primary antibodies. Horseradish peroxidase-conjugated anti-mouse or anti-rabbit antibodies (Cell Signaling Technology, MA, USA) were used as secondary antibodies, and signals were detected by enhanced chemiluminescence (Thermo Fisher Scientific, MA, USA). Images were acquired using Image Lab Software (Biorad. CA, USA).Drug affinity responsive target stability (DARTS) assay

[0322] MDA-MB-231 cells were lysed in mild RIPA buffer with 1% NP40. Cell lysate-drug mixtures were incubated at room temperature on a shaker for 60 minutes to allow binding, and each sample was mixed with 2 pL of pronase solution (Sigma Aldrich)Patent Application Attorney Docket No.: USC-838-PCT (1726)at various dilutions or buffer only (undigested) and incubated at room temperature for 25 min. Protein digestion was stopped by adding protease inhibitor cocktail (Roche), and lysates were mixed with 8 pL of 4x SDS loading buffer and heated at 70 °C for 10 minutes. SDS-PAGE was performed for LOX (ab 174316), LOXL1 and LOXL2 as well as nontargeted control, Cyclin Bl and vinculin.Cell embedding in type I collagen: matrigel (1:1) mix

[0323] ECM with collagen 1 and Matrigel: Collagen solution was prepared at a concentration of 0.5-1 mg / ml from rat tail collagen I (Coming, USA) with a neutralization step including the addition of IN NaOH solution. Cells were trypsinized and resuspended in collagen: matrigel mix (1:1). 8×103cells were seeded into 96-well or p-Slide 8-well glass bottom chambers. After 1 hour of incubation at room temperature, media was added on top of the solidified collagen: matrigel mix. Drug treatments were done 12 hours after cell seeding.

[0324] Production of human foreskin fibroblast (HFF-1) derived ECM: HFF-1 cells were seeded at a density of 120,000 cells / well in a 6-well plate. After day 2, the medium was replaced with fresh medium containing 50 pg / ml of ascorbic acid in every 2 days. At day 6, cells were washed twice with PBS and extracted using the extraction buffer (25 mM NH40H and 0.5% triton X-100 in IX PBS, filtered through 0.45 pM filter). Then, cells were washed again twice with PBS, resuspended in 10 pg / ml DNAse I solution in PBS and incubated at 37 °C for 30 minutes. Lastly, cells were washed twice with PBS slowly.Immunofluorescence staining

[0325] Cells were seeded in p-Slide 8-well (Ibidi, Germany) glass for immunofluorescence staining. Cells were fixed with 4% paraformaldehyde for 15 min, permeabilized and blocked in 3% BSA-PBS. Cells were incubated in Type I collagen and fibronectin primary antibodies and secondary Alexa Fluor 594 or 488-labeled antibodies for 2 hours at room temperature. Cells were also counterstained with DAPI for 5 min. Images were acquired using ZEN Black LSM 880 airyscan (Carl Zeiss, DE). Images were analyzed using the ImageJ software.Lysyl oxidase activity assay

[0326] Cell-based lysyl oxidase activity. To determine cell-based lysyl oxidase activity, fluorometric Lysyl Oxidase Activity Assay Kit (Abeam, USA) was used according to thePatent Application Attorney Docket No.: USC-838-PCT (1726)manufacturer’s instructions. Briefly, 50 pl of supernatant from each well was incubated with reagent mix, and fluorescence was measured in a multimode reader at Ex / Em = 540 / 590 nm wavelength.Recombinant protein-based and conditioned media (CM) based lysyl oxidase activity

[0327] For determining recombinant lysyl oxidase activity’ in initial hit compound screen. Lysyl Oxidase Activity kit (Biovision, USA) was used according to the manufacturer’s instructions. Briefly, LOX (Origene, USA), and LOXL2 (Abeam, USA) recombinant proteins were incubated with the reagent mix in the presence of inhibitors or vehicles. Lysyl oxidase activity was measured in the multimode reader (every 10 minutes, total 110-170 minutes) in fluorescence mode.

[0328] For recombinant protein- and conditioned media (CM)-based lysyl oxidase activity’, Amplex™ Red Hydrogen Peroxide / Peroxidase Assay Kit was used according to the manufacturer’s instruction. Briefly, HEK293T cells were transfected with hLOX and hLOXL2 vectors. At 70-80% confluence, media of the cells were replaced with serum and phenol red-free media containing 0.1% BSA, NEAA and copper sulfate (10 pM for LOX and 100 pM for LOXL2) for 48 hours of incubation. Following the centrifugation of CM, CMs were further 20x concentrated and buffer exchanged to lysyl oxidase assay buffer (1.2 M urea, 50 mM sodium borate buffer, pH 8.2) via 10,000K amicon-centrifugal filter. The concentrated CMs were aliquoted and stored at -80 °C. LOX and LOXL2 CMs or purified LOX (Genscript, USA), LOXL1 (Rockland, USA) and LOXL2 (Genscript, USA) recombinant proteins were prepared in 40 pl of assay buffer and pre-incubated with lysyl oxidase inhibitors in 40 pl with various concentration for 30 min at 37 °C in black plate. After the incubation, 20 pl of Amplex red detection mixture were added into the wells for final concentration as 10 mM of Cadaverine dihydrochloride (or Putrescine dihydrochloride), 100 pl of 10 U / ml HRP. The fluorescence was measured at 60-90 min and fluorescence readings were subtracted from the inhibitor containing buffer blanks without recombinant protein, and lysyl oxidase activity was calculated relative to vehicle containing control.Substrate competition and jump dilution assay

[0329] In substrate competition assay, LOX and LOXL2 CMs were challenged with various doses of Cadaverine dihydrochloride after pretreatment with LXG6403. Lysyl oxidase activity was measured as described above.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0330] In the jump dilution assay, LOXL2 protein was incubated with lOx ICso, 30x ICso and 50x ICso for 30. 60. and 120 min at 37°C. After the incubation, protein-inhibitor mixture was diluted 50-fold in lysyl oxidase assay buffer followed by the addition of Amplex Red reaction mixture containing 10 pM of Cadaverine dihydrochloride as substrate. Lysyl oxidase activity was measured at 90 minutes in the fluorescence mode of the multimode reader.Chemosensitization in 3D culture

[0331] Cells were embedded in type I collagen as described above. Cells were grown in the presence of doxorubicin, cisplatin, or paclitaxel with or without LOX family inhibitors for 48 hours. After incubation with the LOX family inhibitors, cell viability was measured by using 3D Cell Titer Gio (Promega, USA) kit. For the rescue experiments with wt LOX, the LOX ORF containing vector was designed by Vector Builder.ROS measurement

[0332] Cells were pretreated with 20 pM LXG6403 for 24 hours and then embedded in type I collagen as described above and seeded in 24-well plate. For siRNA experiments, cells were transfected with 60 nM siLOX 24 hours before collagen embedding. The next day, cells were treated with 60 pM cisplatin with or without LXG6403 for 48 hours. After 48 hours, cells were washed with PBS and treated with 1.5 mg / ml collagenase solution for 5 min at 37 °C. Cells were dissociated from collagen by pipetting and washed with PBS once. Cells were resuspended in culture media with 10 pM MITOSOX solution and stained at 37°C for 1 hour. Stained cells were transferred to clear bottom black plates with 50 pL per well, and fluorescence was measured in a multimode reader at 488 / 510 nm.Cryosections of the freshly frozen tumors were cut at 5 pm thickness onto the microscope slide. Then, slides were stained with 10 pM of DCFDA for 20 min at room temperature and w ashed with PBS. Upon staining, images were acquired using ZEN Black LSM 880 airyscan (Carl Zeiss, DE). Images were analyzed using the ImageJ software.Real Time Cell Analyzer (RTCA) migration assay

[0333] RTCA CIM Plates were used to assess the migration capacity of the cells upon LXG6403 treatment according to the manufacturer's instructions. Briefly, 160 pl of 10% FBS containing media was added into each bottom chamber / 50pl of FBS free media into the upper chamber and incubated for 1 hour at 37 °C. After the background measurement,Patent Application Attorney Docket No.: USC-838-PCT (1726)5×104cells were seeded in FBS-free media into the upper chamber in the presence of LXG6403 or vehicle. After the 30 min incubation at room temperature, real time cell migration was recorded for 48 hours.MAO-GLO assay

[0334] MAO-GLO assay (Promega, USA) was used to measure the MAO-A and MAO-B activity upon LOX inhibitor treatment according to the manufacturer’s instructions. For the enzyme activity, the substrate was used at a concentration of 160 pM and 16 pM for MAO-A and MAO-B, respectively. MAO-A and MAO-B enzymes (Active Motif, USA) were used in 0.2 pg / per well. Reaction mix, enzymes, and drugs (30, 10, 3, 1.0.3. and 0.1 nM) were mixed and incubated for 3 hours at RT. Then, 50 pl of luciferin was added into wells and incubated for 20 min at RT. Luciferase signals were measured by using a multimode reader.PDX-derived TNBC organoids

[0335] TNBC PDX organoids were established from a freshly frozen surgical tissue by cutting the TM01278 PDX tumors into small pieces and incubating in collagenase A solution with ROCK inhibitor on a shaker at 37 °C for 30 minutes. The collagenase activity was inhibited by adding FBS, and pipetting was done to ensure the formation of almost a single-cell solution. After several washes with PBS. the cell pellet was dissolved in Matrigel. Breast organoid media containing ROCK and GSK inhibitors was added after the Matrigel solidified. For drug testing studies, organoids were disrupted to single cells by digesting at 37 °C for 30 mins with TrypLE (A1217701, Gibco, NY, USA) in the presence of lOuM Rock inhibitor (si 049, Selleckchem, TX, USA). Organoids were then plated into wells of 96 well plates on a Matrigel: collagen 1 (1:1) coated surface with media containing 2% Matrigel (356252, Coming, NY, USA). Drugs were added 72 hours after plating. Organoids were grown in the presence of drugs for 9 days. Organoid viability was measured by using 3D Cell Titer Gio (Promega, USA) kit.Picrosirius red staining

[0336] Picrosirius red staining of paraffin-embedded sections was done using Picrosirius Red Stain Kit from Abeam using the manufacturer’s instructions. Briefly, paraffin sections were melted at 60 °C for 10 min and soaked in Xylene two times for 5 min each, followed by soaking in 100%, 95%, 80%, and 70% ethanol for 2 min each. Then,Patent Application Attorney Docket No.: USC-838-PCT (1726)the slides were rinsed in distilled water and soaked in Picrosirius Red Solution for 1 hour. Rinsing was done in Acetic Acid solution twice for 1 min each. Slides were rinsed in absolute alcohol and mounted in synthetic resin. Slides were imaged under polarized microscope. For image analysis, Hue-Saturation-Balance (HSB) thresholding was applied using the following cut-offs corresponding to the red signal:200 > H < 240 | 150 > S < 255 | 60 > B < 255. 5 different areas per tumor were selected with 3 tumors per group.Total collagen assay

[0337] Total Collagen Assay (Abeam) was performed following the manufacturer's instructions. Briefly, HFF-1 cells were seeded into 6 well plate and at day 2, cells were treated with or without LXG6403 for 4 more days. At day 6, wells were decellularized and HCC1143 cells were seeded on top of the HFF-l-derived ECM. The next day, cells were treated with or without LXG6403 for 48 hours more. Then, cells were solubilized in acid / pepsin mixture (0.5 M acetic acid and 0.1 mg / ml of pepsin) overnight at +4 °C. In soluble fraction, the crosslinked collagen was collected after centrifugation. Finally, the insoluble pellet was dissolved in 10 N NaOH at 120 °C for 1 hour followed by neutralization with 10 N HC1. Oxidized collagen was detected via the kit's reagent. For detection of insoluble collagen in TNBC PDX tumor samples, a small piece from the samples was cut. weighed, and sonicated. Then, samples were solubilized in acid / pepsin mixture (0.5 M acetic acid and 0.1 mg / ml of pepsin) overnight at +4 °C and the rest of the protocol was carried out.Pharmacokinetic (PK) analysis

[0338] Six-to-eight-week-old female NSG (for PDX experiments) or BALB / c (for PK experiments) mice (n=3 per group) were housed in a temperature-controlled and 12-hour light / 12-hour dark cycle environment. PK profde of lead LOX inhibitor, LXG6403 was tested in BALB / c mice. Three mice per group were dosed by oral gavage at 25 or 50 mg / kg. Blood samples were collected by retro-orbital bleeding at time points: 0.25. 0.5, 2, 7 and 24 hours and serums were collected via centrifugation at 4000g for 20 min.LXG6403 was diluted in the serum of healthy mice as the standard, and serum levels of LXG6403 were quantified by LC-MS / MS method. Upon normalizing to the standard readings, the PK parameters, including Cmax, AUC, T1 / 2 values, clearance (Cl) and oral bioavailability (F%) were calculated.Patent Application Attorney Docket No.: USC-838-PCT (1726)Mass spectrometry imaging via MALDI FT-ICR

[0339] Tissue sections were prepared for collagen proteomic imaging followed by sequencing. Imaging data was collected using a MALDI Fourier transform-ion cyclotron resonance (ICR) mass spectrometer (scimaX, Bruker) in positive ion mode with a m / z range of 544.55-2500 and 200 laser shots per pixel. The transient length was 1.3282 s, and the estimated resolving power was 360,000 (at a m / z of 400). A peak of 1570.6768 m / z corresponded with the internal standard. The resulting data were imported into SCiLS Lab 2022 (Bruker), and the peaks were normalized to total ion current (TIC). Sequence data was matched by high mass accuracy ± 8 ppm. A feature table was generated of maximum peak intensities and exported to Excel and GraphPad (Prism v9) where they were further analyzed and statistically evaluated.Collagen sample preparation for MALDI-MSI and MALDI FT-ICR, and data analyses

[0340] Slides containing sections from three different tumors from the vehicle, doxorubicin, LXG6403, and their combination were prepared for imaging of collagen and other extracellular matrix-associated proteins via collagenase type III digestion. Briefly, tissue was cleared using washes of xylenes (twice for three minutes each), 100% ethanol (twice for a minute each), Camoys solution (three times for three minutes each), 100% ethanol (one minute), 95% ethanol (one minute), and 70% ethanol (one minute) followed by water incubation. Tissue was deglycosylated following standardized protocols, and glycans were cleared by soaking in the following solutions in sequence for 1 minute each with 100% ethanol, 95% ethanol, 70% ethanol, water, high pH buffer (10 mM tris buffer, pH 9), water, low pH buffer (10 mM citraconic anhydride buffer, pH 3), and a water rinse followed by drying in a desiccator. Antigen retrieval was completed using published protocols for collagenase digestion, using 10 mM Tris buffer (pH 9) for 20 min at 95°C in a decloaker. The slides were then sequentially diluted with HPLC -grade H2O in three sequential steps for five minutes as they cooled. Slides were dried in a desiccator.Collagenase type III was sprayed using an automated sprayer (M5, HTX Technologies) with a solution of 0.1 pg / pL collagenase type III in 10 mM ammonium bicarbonate with 1 mM CaCL. Fifteen passes were made with an offset of 2.5 mm at 40 °C, 10 psi, and 25 pL / min. Digestion occurred post-spraying by incubating slides in a pre-heated humidity chamber for 5 hours at 37.5 °C. Following incubation, matrix in the form of a-cyano-4-hydroxy cinnamic acid (CHCA, Millipore) was prepared at a concentration of 7 mg / mL inPatent Application Attorney Docket No.: USC-838-PCT (1726)50% acetonitrile, 1% trifluoroacetic acid (TFA). An internal standard of [Glu’]-Fibrinopeptide B human (Sigma) was added to the mixture. The resulting matrix was sprayed using the same parameters listed above with the following exceptions: 14 passes, 79 °C, and 0.07 ml / min. Slides were allowed to dry and then were dipped into 5 mM of cold ammonium phosphate, monobasic and dried upright in a desiccator.

[0341] For mass spectrometry imaging via MALDI FT-ICR, tissues were then scraped into individual centrifuge tubes and resuspended in 10 mM ammonium bicarbonate with 3 mM CaCh. Samples were then sonicated for 30 minutes and then ultrasonicated to further break down tissue particles. Deglycosylation with 1 pg PNGase F was performed overnight at 38 °C and 450 rpm on a benchtop shaker. Samples were again sonicated for 30 min and then spun down for 10 min at 14000 x g at 4 °C. The supernatant was removed, and the pellet resuspended in 10 mM ammonium bicarbonate with 3 mM CaCb. Tissues were then digested twice overnight with 3 pg collagenase type III as indicated previously. The supernatant from digestion was spun down in a speed vacuum to dry down. Intact enzyme and salts were removed using Cl 8 StageTips (Thermo Fisher Scientific). After peptide elution, samples were once again dried down in a speed vacuum, resuspended in 30 pL 0.1% TFA and vortexed and samples of the same treatment were then combined into one tube. C18 Ziptips (2 pg capacity, EMD Millipore) w ere used to Peptides were separated and analyzed on an EASY nLC 1200 System (ThermoScientific) in line with the Orbitrap Fusion Lumos Mass Spectrometer (ThermoScientific) with instrument control software v.4.2.28.14. Two pg of peptides were pressure loaded onto a Cl 8 reversed phase column (Acclaim PepMap RSLC, 75 pm x 50 cm (2 pm, 100 A) ThermoFisher cat. # 164941) and separated using a gradient of 0-40% B in 120 min (Solvent A: 5% acetonitrile, 0.2% formic acid; Solvent B: 80% acetonitrile, 0.2% formic acid) at a flow rate of 300 nL / min using a column heater set to 45 °C. Mass spectra were acquired in data-dependent mode with a high resolution (60,000) FTMS survey scan, a mass range of m / z 375-1575, followed by tandem mass spectra (MS / MS) of the most intense precursors with a cycle time of 3 s. The automatic gain control target value was 4.0e5 for the survey MS scan. HCD fragmentation was performed with a precursor isolation window of 1.6 m / z, a maximum injection time of 40 ms, and HCD collision energy of 35%. The MS / MS scan was acquired at 15,000 resolutions.Multi-photon second harmonic generation (MP-SHG) microscopyPatent Application Attorney Docket No.: USC-838-PCT (1726)

[0342] Fibrillar collagen was imaged in unstained specimens by multiphoton second-harmonic generation (MP-SHG) microscopy. Freshly frozen TNBC PDX tumors were cut as cryosections and fixed with formalin. Then, slides were mounted, and images were taken under 10x 0.40 NA dry objective with z-stack and tiling to image approximately 40-50% of the tumor cross-section via Leica upright DM6000 CFS SP8 multiphoton laser scanning confocal microscope. The tumor slides were scanned with incident light emitted by a Ti: Sapphire femtosecond laser (Coherent Chameleon Ultra II 680-1080 tunable laser) tuned to a wavelength of 880 nm. RLD HyD detectors at 460 / 20 nm captured the second-harmonic signal.

[0343] Fiber width analysis: In Imaged, a 3x3 grid was imposed over the images and each line was analyzed using the Plot Profile function. The software gives a signal intensity vs. pixel position histogram, and the diameter of the fiber bundle was determined as the width of the histogram peaks.Human and mouse liver microsome stability assay

[0344] Human and mouse liver microsome (HLM and MLM) stability assays were performed by Eurofins Panlabs (USA). Briefly, in vitro intrinsic clearance (CLint) of LXG6403 was tested at 2 pM concentration and Imipramine, propranolol, terfenadine, and verapamil were used as control. Samples were incubated for 0, 15, 30, 45, and 60 min, and the remaining compounds were detected by HPLC-MS / MS method.Limited proteolysis experiment

[0345] LOXL2 recombinant protein was used in the limited proteolysis (LiP) experiment to identify the potential interaction regions of LXG6403. Briefly. 20 pg of LOXL2 recombinant protein was incubated with vehicle or 10 pM of LXG6403 for 30 min at 37 °C. Following the incubation, proteinase K treatment (1:100) was done for 5 min at 25 °C and samples were reduced and alkylated using TCEP (5 mM) for 40 min at 37 °C and using IAA (20 mM) for 30 min at 25°C, respectively. Trypsin digestion (1:100) was done overnight at 37°C and trypsin was neutralized with 1% formic acid. Samples were stored at +4 °C until mass spectroscopy.

[0346] Peptides generated through the digestion protocol were separated on a Vanquish LC (Thermo) using a 100 by 2.1 mm X-Bridge Cl 8 column (Waters). The elution solvents were 0.1 % formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The gradient started at 5% B for 5 minutes then raised to 45% B by 40 minutes and 95 %B by 50 minutesPatent Application Attorney Docket No.: USC-838-PCT (1726)then held for a final run time of 60 minutes. Mass spectrometry analysis was performed with a Q-Exactive HFX (Thermo) with electrospray ionization. MSI scans were collected from mass 330 to 1800 interspersed with data dependent (DDA) MS2 scans of the top 10 most abundant peptides. MS raw data was analyzed using MaxQuant (Max Plank Institute) to identify all peptides generated through proteolytic digestion and exported into excel. Non-tryptic peptides were found by sorting the data according to the last peptide residue and filtering out those that did not end in Lys (K) or Arg (R). Non tryptic peptides from inhibitor free and inhibitor (LXG6403) were mapped separately onto the AlphaFold LOXL2 structure by coloring according to free or bound and these structures then overlayed to identify regions that LXG6403 did not interact with.Bioinformatics analyses

[0347] A list of gene names that appeared in the proteome determined to be significantly different between treatments (based on FDR-dependent student's t-tests in Perseus) was entered into the DAVID (Database for Annotation, Visualization, and Integrated Discovery) search tool to run against a human Homo sapiens) database to annotate and investigate cellular processes potentially associated with this particular group of genes. Functional annotation clustering was performed on the submitted list, which resulted in 11 different annotation clusters, each given an enrichment score based on the geometric mean of each member of the annotation’s p-value in -log scale. Each annotation contains relevant terms and phrases associated with biological processes or organelles based on the shared number of genes from the list. The associated -val ues were determined based on Fisher Exact Test / EASE score. For MALDI-MSI analysis, the monoisotopic-precursor selection was set to “peptide”. Precursors within 10 ppm mass tolerance were dynamically excluded from resequencing for 25 sec. Advanced peak determination was enabled. Precursor ions with charge states that were undetermined or >7 was excluded. MaxQuant v 2.0.1.0 was used to search MS / MS data, remove contaminants and target oxidation of methionine and proline and hydroxylation of prolines as post-translational modifications (PTM). Perseus v1.6.15.0 was used to further analyze the proteomics data. Peptide intensities were log2 transformed and then normalized via median subtraction. Peptide identities were assigned to accurate mass image data by accurate mass matching < 5ppm. Volcano plots were generated based on student’s t-tests with an FDR = 0.05 and SO = 3. Heatmaps of normalized MS / MS data were generated. Clustering was done using One minus person correlation metric and average linkage method.Patent Application Attorney Docket No.: USC-838-PCT (1726)Cd #ompounQuantification and Statistical Analysis

[0348] All the results are represented as mean ± standard deviation (SD) or mean ± standard error of the mean (SEM), as indicated in the figure legends. All statistical Sttrucureanalyses were performed in GraphPad Prism Software. Differences were assessed using the one-tailed or two-tailed unpaired or paired Student’s t-test or one-way ANOVA, as indicated in the legends. Tumor volume or body weight changes over time between combination groups vs. single agent or vehicle-treated groups were compared using two-way ANOVA with Dunnett multiple comparison test. Groups were compared with Mann Whitney U tests for the fiber width analysis using MP-SHG. The differences were considered statistically significant iff’ < 0.05 and indicated by * or P < 0.01 and indicated by ** unless otherwise stated as in the legends.Table 1. LOX Family Inhibitors.R1 R2 R3 R4 R5 R6 R76415 1 CH3 H CH3 H H H H6403 1 CH3 H H H H NH(C=O)CH3 H1 6487 2 CH3 H H H H H NH(C=O)CH3 2 6506 2 CH3 COCH3 H - H H NH(C=O)CH3 3 6425 1 CH3 H H H H NH(C=O)CH2C H H2OCH34 6512 1 CH3 H H H NH(C=O)CH3 H H5 6488 1 CI 13 II II II II5'-methyl-[4,4- 6 6532 1 CH3 H H H H bithiazole]-2,2'- H diamine(C=O)NH-3-F- 7 6578 1 CH3 H H H H HPh8 6576 2 CH3 H H H (C=O)NH-CH3 H H NH(C=O)-4- 9 6558 1 CH3 H H H H pyridyl H10 6574 2 CH3 H H H CH=CH- N=CH- H11 6539 1 CH3 H H H H (C=O)N, N-di- H CH3NH(C-O)CH2C12 6426 1 CH3 H H H H H H3CH2CH313 6489 1 CH3 H H H HPatent Application Attorney Docket No.: USC-838-PCT (1726)14 6505 1 CH3 COCHS H H H NH(C=O)CH3 H15 6519 1 CH3 COCH3 co H H CH316 6522 1 CH3 H H H H H H17 6523 1 CHS H H H H NH(C=O)Ph H NH(C=O)CH2- 18 6524 1 CH3 H H H H H 4ClPh19 6528 1 CH3 H H H H NH(C=O)-2.4- Hdi-F-PhNH(C=O)CH2- 20 6529 1 CH3 H H H H H4-F-Ph21 6530 1 CH3 H H H H (C=O)NH-Ph H22 6531 1 CH3 H H H H COOH H NH(C=O)Ph- 23 6533 1 CH3 H H H H H NH(C=O)CH3NH(C=O)-4- 24 6577 1 CH3 H H H H H OMe-Ph26 6538 1 II II II - - -- - 27 6554 1 CH3 H H H H (C=O)NH-CHS H (C=O)NH-Ph- 28 6555 1 CH3 H H H H H NH(C=O)CH3(C=O)- 29 6556 1 CH3 H H H H piperazine- H Nmethyl(C=O)NH- 30 6559 1 CH3 H H H H H 4pyridyl31 6560 1 CH3 H CH3 H H COOH H NH(C=O)-Ph- 32 6561 1 CH3 H H H H H NH(C=O)CH3NH(C=O)-Ph- 33 6562 1 CHS H H H H H2,4 difluoro5'-methyl-[4,4- 34 6563 1 CH3 H H H H bithiazole]-2,2'- H diamine35 6568 1 CHS H H H COOH H H36 6569 1 CH3 H H H (C=O)NH-CHS H H37 6570 1 CH3 H H H (C=O)NH-Ph H H38 6571 1 CHS H H H (C=O)NH-Ph- H H NH(C=O)CH3NH(C39 6572 1 CHS H H =O)C H H HH3pyrimidinyl40 6573 1 CH3 H H H H (3,5) H41 6575 3 CH3 H H CO2Et H H H42 6609 1 CH3 H H H CH2- CO-NH- H43 6606 1 CHS H H H CO-NH- CH2- H44 6624 1 CHS CH2-Ph H H CH2- (C-O)-NH- HCH2- 45 6603 1 H H H NH(C=O)-CH3 H HCH3Patent Application Attorney Docket No.: USC-838-PCT (1726)46 6663 1 CH3 H CH3 H H NH(C=O)CH3 H47 6635 1 CH3 H H H NH(CO)- CH2- H48 6678 2 CH3 H CH3 H H H (C=O)NH-CH3 50 6604 1 CH3 CH2-CH3 H H NH(C=O)-CH3 H H51 6619 1 CH3 H CH3 H NH(C=O)-CH3 H H52 6614 1 CH3 H H H NH(C=O)-CH3 Cl H53 6616 1 CH3 H H H Cl NH(C=O)-CH3 H NHCOCH2CH254 6592 1 CH3 H H H H H OMe55 6605 1 H H H H NH(C=O)-CH3 H H56 6681 1 CH3 H H H (C=O)NH-CH3 Cl HH (centralthiazole replaced57 6613 1 CH3 H H H NH(C=O)-CH3 Hwith Imidazolering)58 6677 1 CH3 H CH3 H NH(C=O)CH3 Cl H59 6626 1 CH3 CH2-Ph H H CH2- (C=O)-NH- H61 6634 1 CF3 H H H CH2- (C=O)-NH- H62 6636 1 CH3 H H H CH2- NH(C=O)- H NH(C=O)-Ph- 63 6621 1 CH3 (C=O)Ph H H NH(C=O)CH3 H H NHCOCH2CH264 6622 1 CH3 (C=O)Ph H H H H OMe65 6580 3 CH3 H H C02H H -- -- 67 6639 1 CH3 H H H CH2- (C=O)-N-CH3 H68 6638 1 CH3 H H H (C=O)-N-CH3 CH2- H CONH(2,4- 70 6591 2 CH3 H H H H Hdifluorophenyl) 71 6637 1 CH3 CH2-CH3 H H NH(SO2)-CH3 H H72 6579 2 CH4 H H H H H CO2H73 6607 1 CH3 CH3 H H NH(C=O)-CH3 H H76 6594 2 CH3 H H H CH=CH- HN-C=O H77 6631 1 CH3 H H H (C=O)- (C=O)-N-CH3 H79 6641 1 CF3 H H H NH(SO2)-CH3 H H80 6582 2 CH3 H H H H H CONHPh CONH(4-NHAc- 84 6581 2 CH3 H H H H H Ph)87 6625 1 CH3 H H H (C-O)- (C-O)-NH- H88 6595 2 CH3 (C=O)CH3 H H CH=CH- N=CH- H89 6628 1 CF3 H H H NH(CO)-CH3 H H (C=O)NH-(CH2)2- 94 6657 2 CI 13 II II II II IICH395 6658 2 CH3 H H H H H (C=O)NH-(CH2)4- PhPatent Application Attorney Docket No.: USC-838-PCT (1726)96 6664 1 CHS H CH3 H H (C=O)NH-CH3 H98 6679 1 CHS H CHS H CH2- (C=O)NH- H99 6680 1 CH3 H CH3 H (C=O)-NH-CH3 H H101 6670 1 CH3 H H H CH2-NH2 H H108 6698 1 CHS H CH3 H CH2-NH2 H H109 6699 1 CHS H CHS H H CH2-NH2 H110 6700 1 CHS CH2-NH2 H H (C=O)NH-CH3 H H111 6701 1 CH3 CH2-NH2 H H H (C=O)NH-CH3 H114 6706 1 CH3 H H H CH2- NMe(C=0)- H6574- C6H4115 2 NH2 H H CH=CH- N=CH- H analog (-13-)6574- C6H4116 2 NH2 CH3 H CH=CH- N=CH- H analog (-1,3-)6681- 117 2 CH3H H H H Cl (C=O)NH-CH3 analog6681- 118 2 CH3H CHS H H Cl (C=O)NH-CH3 analog6574- CH=C-CONH- 119 analog 2 CH3H H H CH3 N=CH- H6574 CH=C-CONH- 120 2 CH3H CH3 H N=CH- H analog CH36574- CH-C-CONH- 121 2 CH3H H H N=CH- Hanalog CH3* Structures 1, 2, 3 refer to Fig. 2.** Ascended sorting of IC50, except three initial lead compounds, 6403, 6487, and 6512Example 1

[0349] A diverse library of new LOXi derivatives was generated via an extensive SAR study using initial pharmacophore (Fig. 1G). The initial expansion library around the biphenyl- [4,5'- bithiazol]-2-amine core structure led to the identification of first-generation lead LOXi inhibiting cellular LOX activity at low pM ICso. A compound library and detailed SAR was generated by making substitutions at positions R1 to R6 already shown to improve activity (Tables 1-2), in addition to the positions that had not been previously modified. In the Phase I study to date, new derivatives of the lead, 6403, has been synthesized and screened, including isosteres for the core bisthiazole component (Tables 1-2).Table 2; Activity of LOX Family Inhibitors.Cell-Based LOXID StructureActivity Normal Cell Toxicity MDA-MB- 231 HCC1143 HFF HEK293T IC50, Viability' IC50, gM Viability IC50, jiM IC50, pM range at 25 jiM range at 25 LilVI 6415 1 1.68 >50 97%6403 1 1.46 1 11 -100 78% 50-100 98% 6487 2 1 1 01 100 83% 50-100 93%6506 2 0.59 1 09 -100 77% -100 98%Patent Application Attorney Docket No.: USC-838-PCT (1726)6425 1 0.58 >100 95% >100 99% 6512 1 0.87 258 -100 85% 50-100 99% 6488 1 0.35 4.2 50-100 94% 25-50 87% 6532 1 0.49 206 50-100 100% 6.2-12.5 4% 6578 1 2.18 1 74 12.5-25 40% 6.2-12.5 5% 6539 1 0.88 -100 94% 25-50 96% 6426 1 1 100 91% 50 91% 6489 1 1 -100 72% -12.5 27% 6505 1 23.3 12.5-25 50% 12.5-25 38% 6519 1 1.22 -12.5 44% -3 13% 6522 1 47 >100 100% 25-50 70% 6523 1 2.43 50-100 89% 125-25 40% 6524 1 >106528 1 3.06 50 73% 6.1-12.5 15% 6529 1 1 86 50-100 50% 125-25 45% 6530 1 1.57 1.98 25-50 60% -12.5 9% 6531 1 1.49 >100 100% > 100 100% 6533 1 6.51 >100 100% >100 90% 6577 1 4.34 >100 100% 25-50 59% 6538 1 >10 0.79 >100 100% >100 100% 6554 1 0.63 2.56 >100 87% 50-100 78% 6555 1 0.81 1.99 >100 100% >100 92% 6556 1 2 86558 1 2.936559 1 2.136560 1 >106561 1 3.686562 1 436563 1 3.896568 1 >106569 1 1.926570 1 6.166571 1 >106572 1 2.976573 2 7.336574 2 1.296575 3 2.026576 2 1 36626 1 2 16634 1 2.146636 1 2.186621 1 2.226622 1 2 36580 3 2.316563 1 2.346639 1 2.496638 1 2.526559 1 2.586591 2 2.78Patent Application Attorney Docket No.: USC-838-PCT (1726)6637 1 2.956572 1 2.976579 2 3.046607 1 3.086594 2 4.056631 1 4.366641 1 4986582 2 6.066570 1 6.166640 1 6.346581 2 7.076573 1 7.336568 1 11 386625 1 14.76560 1 14786595 2 16446571 1 16536562 1 22456628 1 >106657 386658 576664 616665 836679 3.66680 276670 376698 846699 12.46700 66701 10.36698 846706 0.46

[0350] We began by deconstructing the molecular framework of 6403 (initial lead, IC50=1.4 pM. Cetin-Saatci et al, Cell Chem Biol, 2024) to identify critical pharmacophores. Compound 6538, with only the bithiazole core (excluding the aniline moiety) lowered the activity by 10 folds compared to 6403, showing the necessity of the aniline segment. To evaluate the role of amino substituent (R2) on the bithiazole ring, we synthesized the acylated analog 6505, which resulted in a 7-fold decrease in activity, showing the essentiality of a free amino group at R2. Replacing para-amino group (R7) in 6403 with a meta-amino group (R6) to form 6512 increased the potency (IC50=0.75 pM), indicating that 1,3-diamino substitution is more favorable than 1,4-diamino. Notably, 6606 (an acyl derivative of 6487) showed 2.5-fold increase in activity (IC50=0.57 pM) suggesting that hetero-aromatic frameworks may accommodate such modifications.Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0351] Recognizing the potency of 6512, we synthesized analogs with modifications at Rl. R2, R3, and R5. R3 methylation (6619). R5 (6637, and 6641), and some of the R2 modifications (6607 and 6604) retained comparable activity (0.97-4.98 pM), and reverse amides (6568, 6569, 6570, and 6571) mostly showed reduced activity (6.16-16.13 pM) except 6569 (ICso=1.92 pM). Central ring replacements, imidazole (6613), retained modest activity (IC50=1.65 pM).

[0352] We synthesized multiple heterocyclic derivatives. Pyridine analog 6487 (0.9 pM) was among the most potent compounds in Phase I. Its analogs (6576, 6579, 6581, 6582, and 6591) displayed a wide range of activities (1.3-7.1 pM). Quinoline-based 6574 (1.29 pM) retained potency, while its acety lated derivative 6595 lost activity (16.4 pM). Pyrimidine (6573). and quinolinone (6594) analogs were less active. Tri-thiazole derivatives (6575 and 6580) did not outperform 6487.

[0353] Non-N-methylated phthalimide analogs (6606 and 6609) exhibited improved potency (0.57 and 0.6 pM). However, N-methylation or CF3 substitution led to reduced activity (2.1-14.7 pM). Oxindole (6635 and 6636) derivatives showed different results. 6635 had potent activity (0.93 pM), whereas most R2-modified derivatives were inactive.

[0354] In summary, our comprehensive SAR study revealed several key insights. Both the bithiazole and aniline segments are crucial for LOX inhibition. The position and nature of amine substituents critically influence potency. Replacing phenyl rings with heterocycles or modifying them with specific acyl, alkyl, benzoyl, benzyl or sulfonamide groups enhanced or sustained activity. Specific novel frameworks, such as pyridine, phthalimide, and oxy-indole-based bithiazoles, improved activity and represent promising candidates for further development

[0355] To identify the minimal pharmacophore of 6403, a derivative. 6538, was generated that retains only the bithiazole core but lacks the aniline segment (Tables 1 and 2). The LOX activity inhibition of 6538 was at least 10-fold lower than 6403, indicating that the aniline component is essential for LOX activity inhibition. Replacing the aniline group with hetero-aromatic rings including pyridine (6487) or indole (6488), increased LOX inhibition relative to 6403. Given the importance of both bithiazole and aniline components, the role of R2 amino group on the bithiazole core was then explored. Masking this amine (NH2) group through acetylation (R2, 6505) led to a 15-fold reduction in LOX activity inhibition compared to 6403, showing its importance. It is interesting to note that when R2 was acetylated in the context of 6487 (pyridyl), the resulting acyl-derivative, 6506, had 2.5-fold increased LOX inhibition compared to 6403. Moving the R6 acetamidePatent Application Attorney Docket No.: USC-838-PCT (1726)group in 6403 to R5 (para to meta) to generate 6512 led to a further 2-fold increase in LOX inhibition, relative to 6403 (Table 1). Without wishing to be bound by theory, this suggests that the 1,3-diamino substitution of 6512 (aniline component) is more active than the 1,4-diamino substitution found in 6403. Since our initial 6403-SAR data analyses strongly supported focusing on aromatic / het-aromatic ring modifications for activity studies, the effects of the following three R6 modifications in 6403 was assessed: 1) replacement of NH- acetyl group by NH-benzoyl (6523) and phenylacetamide groups (6524), 2) reverse-amide functionalization (6530 and 6539); and 3) the formation of 1,4-diamino-bis-bithiazole derivative (6532). Compared to 6403, the NH- benzoyl and phenylacetamide had decreased LOX inhibition. The reverse-amide functionalization led to a consistent and measurable increase of LOX inhibition compared to the starting amide derivatives (compare 6403 and 6554, 6523 and 6530). The symmetric bis-bithiazole derivative 6532 showed a ~2.5-fold increase in LOX activity inhibition compared to 6403 (Tables 1 and 2), indicating the benefit of a bifunctional compound. Changing this from the 1-4-phenylene to the 1-3 derivative resulted in a 4-fold loss of activity suggesting the structural requirements of the para substitution for the binding mode. A series of derivatives were synthesized to expand on the promising SAR observed for the R5 substituted compound (6512) which is 2-fold more potent than 6403. 6568-6571 all had lower activity than their R6 counterparts suggesting that the benefit observed with 6512 was unique to this analog. Furthermore, generation of the R4 substituted version of 6403 indicated that while this change was tolerated, it had slightly reduced potency. The reverse amide version of 6487 (6576) showed similar activity to the parent compound. Further analogs incorporated thiazole and quinoline replacement for the aniline ring of the 6403 series and these again had similar activity opening up new scope for structure activity development. In summary, a library of novel LOXi was obtained with equivalent or improved activity to advance in our development pipeline.

[0356] Based on the HTS data, SAR analysis and designed 15 derivatives of compound 1 was performed. The general scheme to synthesize the bi-thiazole analogs, and the functional groups in each analog are shown below.Patent Application Attorney Docket No.: USC-838-PCT (1726)General synthesis scheme of bi-thiazole analogsk I - *- 1^' H 'N'' I - - / - - R^.. H J ] ii HN- / '"•& R3-<> -V^FV..an&ne thiourea “3bi-thsasole derivatives derivatives analogs

[0357] The detailed scheme to synthesize the bi-thiazole analogs is shown below including the synthesis of substituted 4-methylthiazol-2-amine and aniline rings.Synthesis of l-(2-amino-4-methyl-thiazol-5-yl)-2-bromo-ethanone

[0358] l-(2-amino-4-methyl-thiazol-5-yl)ethanone (5 g) was dissolved in HBr (20 mL) and allowed to warm to 60 °C. Bn (4.6 g) was then dissolved in 1,4-dioxane and was added in a dropwise manner. The reaction was allowed to stir at 60 °C for 3 hours. After cooling to RT, the mixture was basified with NarCCh after which solid product came out of solution. The solid was removed by fdtration. After extraction of the remaining solution with ethyl acetate, the organic layer was then washed with brine, dried over sodium sulfate, and was evaporated under reduced pressure to dryness. The remaining crude material was combined with the filtered solid and was dried in a vacuum desiccator, isolated as white solid; 55% yield; 'H-NMR (400 MHz, DMSO-cL) 5 ppm 9.30 (br, 2H), 4.88 (s, 2H), 2.47 (s, 3H);13C-NMR (400 MHz, DMSO-Z) 5 ppm 183.45, 170.27, 169.13, 118.15, 48.95, 16.92; ESI-MS (pos): m / z 235 (M+H)+(expected 235.10).Synthesis ofN-[4-(carbamothioylamino)phenyl]acetamide (compound 9)

[0359] N-(4-aminophenyl)acetamide (6 g) was dissolved in anhydrous DCM and benzoyl isothiocyanate was added in a dropwise fashion at RT. The reaction was then allowed to stir overnight for 16 h. The solvent was removed, and the products re-dissolved in 1: 1 THF / 1M NaOH, and then allowed to reflux at 75 °C for 3 hours. The sample was cooled to RT, diluted in water (-150 mL) and was extracted ethyl acetate (3X50 mL) to yield N-[4-(carbamothioylamino)phenyl]acetamide as white solid; 35% yield; mp 178-180Patent Application Attorney Docket No.: USC-838-PCT (1726)°C; ll-NMR (400 MHz, D SO-cL) 5 ppm 9.94 (s, 1H), 9.61 (s, 1H), 7.52 (d, J= 8.75 Hz, 2H), 7.27 (d, J= 8.60 Hz, 2H), 2.03 (s, 3H);13C-NMR (400 MHz, DMSO-tfe) 5 ppm 181.44, 168.58, 136.62, 134.41, 124.43, 119.73, 24.39; ESI-MS (pos): m / z 210.0309 (M+H)+(expected 210.06).Synthesis of N-[ 4-[[4-(2-amino-4-methyl-thiazol-5-yl)thiazol-2-yl amino ] phenyl Jacetamide (compound 9)

[0360] N-[4-(carbamothioylamino)phenyl]acetamide (1.1 g) and 1 -(2-amino-4-methyl-thiazol-5-yl)-2-bromo-ethanone (1.85 g) were dissolved in 30 mL ethanol and the solution was heated to 90 °C for 2-3 h. After completion of the starting material the solvent was removed under reduced pressure at 45 °C water bath temperature. The crude material was re-dissolved in 3 mL of DMSO and purified by Medium Pressure Liquid Chromatography using a C 18 reverse phase column and acetonitrile-water as eluent with 1 % aqueous formic acid additive to obtain the desired product as brown solid; 41% yield; mp 212-215 °C;1H-NMR (400 MHz, DMSO-cfc) 5 ppm 10.37 (s, 1H), 9.91 (s, 1H), 9.25 (br, 2H), 7.54 (s, 4H), 7.02 (s, 1H), 2.42 (s, 3H), 2.03 (s, 3H);13C-NMR (400 MHz, DMSO-Je) 5 ppml68.39.167.61, 163.80, 140.10, 136.59, 134.24, 131.68, 120.17, 117.94, 113.97, 103.93, 24.35, 13.73. ESI-MS (pos): m / z 346.1247 (M+H)+(expected 346.08).Synthesis of 4-methyl-5-[2-(4-nitroanilino)thiazol-4-yl]thiazol-2-amine (compound 1)

[0361] 1) Synthesis of (4-nitrophenyl)thiourea: 4-nitroaniline (2.0 g) was suspended in 2 mL of dry DCM and the benzoyl isothiocyanate (2.95 g) were separately dissolved in 1 mL of dry DCM. The benzoyl isothiocyanate was then added to the aniline dropwise over the course of 1 min with stirring. The solution was then evaporated under reduced pressure to dryness and dissolved in a solution of THF / 1M NaOH (1: 1). The solution was then refluxed at 75 °C for 1 hour. The sample was then cooled to room temperature, diluted with water, and extracted 3x30 mL with ethyl acetate. The organic solution was then dried over sodium sulfate, filtered, and evaporated under reduced pressure to dryness andPatent Application Attorney Docket No.: USC-838-PCT (1726)purified with medium pressure liquid flash chromatography to yield the product (0.181 g). ESI-MS (pos): wz 198.0 (M+H)+(expected 198.0).

[0362] 2) Synthesis of 4-methyl-5-[2-(4-nitroanilino)thiazol-4-yl]thiazol-2- amine (compound 1): (4-nitrophenyl)thiourea (0.18 g) and l-(2-amino-4-methyl-thiazol-5-yl)-2-bromo-ethanone (0.30 g) were dissolved in ethanol and were allowed to reflux for 90 min after which the sample was then evaporated under reduced pressure to dryness, dissolved in DMSO, and was purified by reverse-phase flash chromatography to yield the product (0.30 g, 99% purity), (expected 334.0). 'H-NMR (400 MHz, Ds Methanol) 5 ppm 8.027-8.124 (m, 3H), 7.742-7.766 (d, 2H), 6.667 (s, 1H), 2.326 (s, 3H). ESI-MS (pos): m / z 334.0 (M+H)+(expected 334.0).Synthesis of N-[ 4-[[4-(2-amino-4-methyl-thiazol-5-yl)thiazol-2-yl ]amino Jphenyl ]-3-methoxy-propanamide (compound 10)

[0363] 1) Synthesis of N-[4-(carbamothioylamino)phenyl]-3-methoxy-propanamide: N-(4-aminophenyl)-3-methoxy-propanamide (0.24 g) was suspended in 2 mL of dry DCM and the benzoyl isothiocyanate (0.26 g) was separately dissolved in 1 mL of dry DCM. The benzoyl isothiocyanate was then added to the aniline dropwise over the course of 1 min with stirring. The solution was then evaporated under reduced pressure to dryness and dissolved in a solution of THF / 1M NaOH (1:1). The solution was then refluxed at 75 °C for 3 hours. The sample was then cooled to room temperature, diluted with water, and extracted 3x10 mL with ethyl acetate. The organic solution was then dried over sodium sulfate, filtered, and evaporated under reduced pressure to dryness and purified with medium pressure liquid flash chromatography to yield the product as white solid; 45% yield; mp 155-157 °C; 'H-NMR (400 MHz, DMSO-tri) 5 ppm 9.94 (s, 1H). 9.55 (s, 1H), 7.54 (d, J= 8.76 Hz, 2H), 7.26 (d, J= 8.68 Hz, 2H), 3.61 (t, J= 6.20 Hz, 2H), 3.24 (s, 3H) 2.53 (t, J= 6.16 Hz, 2H);13C-NMR (400 MHz, DMSO-rie) 5 ppm 181.43,Patent Application Attorney Docket No.: USC-838-PCT (1726)169.55, 136.53, 134.43, 124.49, 119.76, 68.65, 58.39, 37.43; ESI-MS (pos): m / z 254.1 (M+H)+(expected 254.1).

[0364] 2) Synthesis of N-[4-[[4-(2-amino-4-methyl-thiazol-5-yl)thiazol-2-yl] amino] phenyl]-3-methoxy-propanamide (compound 10): N-[4-(carbamothioylamino)phenyl]-3-methoxy-propanamide (0.019 g) and 1 -(2-amino-4-methyl-thiazol-5-yl)-2-bromo-ethanone (0.023 g) were dissolved in ethanol and were allowed to reflux for 90 min after which the sample was then evaporated under reduced pressure to dryness, dissolved in DMSO, and was purified by reverse-phase flash chromatography to yield the product as brown solid; 35% yield; mp 187-189 °C; 'H-NMR (400 MHz, DMSO-dfi) 5 ppm 10.40 (s, 1H), 9.95 (s, 1H), 9.30 (br, 2H) 7.59-7.52 (m, 4H), 7.03 (s, 1H), 3.61 (t, J= 6.20 Hz, 2H), 3.24 (s, 3H) 2.54 (t, J= 6.20 Hz, 2H), 2.42 (s, 3H);13C-NMR (400 MHz, DMSO-cL) 5 ppm 169.31, 167.59, 163.81, 140.05, 136.64, 134.15, 131.51. 120.17, 117.95, 113.93, 104.03. 68.70, 58.38. 37.37, 13.68; ESI-MS (pos): m / z 390.1 (M+H)+(expected 390.1).Synthesis ofN-[4-[[4-(2-amino-4-methyl-thiazol-5-yl)thiazol-2-yl]amino]phenyl]butanamide (compound 11)H

[0365] 1) Synthesis of N-[4-(carbamothioylamino)phenyl]pentanamide: N-(4-aminophenyl)pentanamide (0.044 g) was suspended in 2 mL of dry DCM and the benzoyl isothiocyanate (0.047 g) was separately dissolved in 1 mL of dry DCM. The benzoyl isothiocyanate was then added to the aniline dropwise over the course of 1 min with stirring. The solution was then evaporated under reduced pressure to dryness and dissolved in a solution of THF / 1M NaOH (1:1). The solution was then refluxed at 75 °C for 3 hours. The sample was then cooled to room temperature, diluted with water, and extracted 3x10 mL with ethyl acetate. The organic solution was then dried over sodium sulfate, filtered, and evaporated under reduced pressure to dryness and purified with medium pressure liquid flash chromatography to yield the product as white solid; 41% yield; mp 145-147 °C; ‘H-NMR (400 MHz, DMSO-tL) 5 ppm 9.85 (s, 1H), 9.54 (s, 1H), 7.54 (d, J =Patent Application Attorney Docket No.: USC-838-PCT (1726)8.76 Hz, 2H), 7.25 (d, J= 8.64 Hz, 2H), 2.29 (t, J= 7.36 Hz, 2H), 1.57 (m, 2H) 1.30 (m, 2H), 0.89 (t, J= 7.32 Hz, 3H);13C-NMR (400 MHz, DMSO-t / e) 5 ppm 181.42. 171.60, 136.69, 134.29, 124.51, 119.78, 36.55, 27.76, 22.27, 14.19; ESI-MS (pos): m'z 252.1 (M+H)+(expected 252.1).

[0366] 2) Synthesis of N- [4- [ [4-(2-amino-4-methyl-thiazol-5-yl)thiazol-2-yl] amino] phenyl] butanamide (compound 11): N-[4-(carbamothioy lamino) phenyl]pentanamide (0.0073 g) and l-(2-amino-4-methyl-thiazol-5-yl)-2-bromo-ethanone (0.0047 g) were dissolved in ethanol and were allowed to reflux for 90 min after which the sample was then evaporated under reduced pressure to dryness, dissolved in DMSO, and was purified by reverse-phase flash chromatography to yield the product as brown solid; 37% yield; mp 180-183 °C; 'H-NMR (400 MHz, DMSO-cL) 5 ppm 10.39 (s, 1H), 9.87 (s, 1H), 9.30 (br, 2H) 7.58-7.51 (m, 4H), 7.02 (s, 1H), 2.42 (s, 3H), 2.29 (t, J= 7.36 Hz, 2H), 1.56 (m, 2H) 1.30 (m, 2H), 0.89 (t, J= 7.28 Hz. 3H);13C-NMR (400 MHz. DMSO- e) 5 ppm 171.44, 167.60, 163.84, 140.04, 136.54, 134.28, 131.50, 120.22, 1 17.96, 113.94, 104.00, 36.52, 27.82, 22.30, 14.22, 13.67; ESI-MS (pos): m / z 388.1 (M+H)+(expected 388.1).Example 2

[0367] To identify potent and safe LOX family inhibitors, a high-throughput screen (HTS) with a cell-based LOX activity assay was performed with more than 5,000 drug-like compounds from the EXPRESS-Pick diversified library (4,480 molecules, ChemBridge) and Tocriscreen library of bioactive compounds (1,280 compounds, Tocris), each at a single concentration of 10 pM using the MDA-MB-231 TNBC cell line (Figs. 1A-1B). This screen identified 25 compounds from the ChemBridge library and 56 hits from the Tocris library that successfully inhibited > 90% of cellular LOX activity. As such, the primary’ focus was to identify cell-permeable molecules with minimum cellular toxicity, toxic molecules were eliminated by counter screening with a cell cytotoxicity assay. Here, compounds that inhibit LOX activity' >75% (64 from ChemBridge and 64 from Tocris) were screened for LOX activity' and cell viability' inhibition simultaneously (Figs. IB and 7). The most potent hits (20 in total) that inhibit LOX activity without major cytotoxicity were then verified by re-ordering fresh compounds and testing them in the cell-based LOXPatent Application Attorney Docket No.: USC-838-PCT (1726)activity and cell viability assays (Fig. 1C). Of these 20 compounds, five of them were shortlisted based on their ability to sensitize TNBC cells to doxorubicin in 3D collagen I culture (Fig. ID and 7), which aligns with previous observations that LOX inhibition overcomes doxorubicin resistance. Five compounds that induced doxorubicin sensitization were further tested in recombinant LOX and LOXL2 protein-based activity assays (without preincubation) to determine their on-target activity and selectivity (Figs. 1E-1F). Two of these compounds, 1 and 2 strongly inhibited the activity of recombinant LOX (Fig. IE), and moderately inhibited the acti vi ty of LOXL2 at 10 pM, a concentration that is 1000X lower than required for BAPN (Fig. IF), which is commonly used LOX family inhibitor in in vitro and preclinical models despite its toxicity in clinic.

[0368] Of these two compounds, 1 (R4= NO2. Table 1) met all our stringent criteria including synthetic feasibility and favorable drug-like properties and was therefore selected for SAR and initial lead optimization. For this purpose, a diverse set of analogs of 1 (phenyl bithiazole diamine core structure) were commercially obtained or synthesized to increase potency and selectivity. Several analogs possessed similar or increased activity relative to 1, identified the sensitive positions on the core molecule and thus generated insights into the SAR (Table 2). A compound generated with an acetamide group at R1 showed decreased activity relative to the free amino (6 vs. 1), thus highlighting the importance of this group on the terminal thiazole ring and suggesting that it is an important binding determinant for this series. Replacement of the nitro group (R4) with an amine (7) resulted in loss of activity thus showing that less favorable interactions result in this compound. Substitution of this amine through dimethylation and acylation recovered LOX activity, indicating favorable contacts of the modified analogues (8, 9, 10, and 11).Replacement of the nitro group of 1 with hydroxyl (12). alkoxy (14, 15 (IC50: 12.5 pM)) and with sulfonamide (16) was well tolerated. N-methylation of the aniline nitrogen in the 4-hydroxy context to generate a tertiary amine (13) resulted in potency increases relative to the activity of the parent compound (12). Substitution of the aniline ring at the 2 and 3 positions (R2, R3) diminished LOX activity with exception of the dimethoxy analog (20).Among all tested analogs, compound 9 inhibited the cellular lysyl oxidase activity most efficiently in MDA-MB-231 cells with an IC50 of 1.3 pM (Tables 1-2). As such, compound LXG6403 was selected as the lead compound.

[0369] To assess direct inhibition of LOX, the inhibition modality, and the specificity' of lead compound LXG6403, a series of experiments were performed using commercially available recombinant LOX family proteins as well as concentrated tagged LOX familyPatent Application Attorney Docket No.: USC-838-PCT (1726)proteins from the conditioned media of HEK293T mammalian cells overexpressing HA-tagged LOX proteins. Firstly, we determined the ICso of LXG6403 against recombinant LOX (purified from CHO-S), LOXL2 (purified from CHO-S) and LOXL 1 (purified from Baculovirus infected Sf9 cells) proteins. As shown in Figs. 2A-2D, LXG6403 was identified to be slightly more selective towards LOX (~3.5 fold) compared to LOXL2 (ICso = 0.28 pM for LOX and 0.95 pM for LOXL2), while it did not inhibit LOXL1 (ICso > 10 pM). Here, we used BAPN as a positive control that inhibits the activity of all recombinant proteins (Fig. 8A). Next, LOX and LOXL2 proteins were concentrated from the conditioned media of HEK293T mammalian cells overexpressing HA-tagged LOX (rLOX-CM) or LOXL2 (rLOXL2-CM) for subsequent characterization of our inhibitor (Fig. 8B).This is a widely used, feasible method to obtain large amounts of active LOX proteins to study effects of newly developed inhibitors. In line with results herein using recombinant LOX and LOXL2 proteins, LXG6403 was again found to be slightly more selective (~3-fold) towards rLOX-CM compared to rLOXL2-CM (ICso = 0.95 pM for LOX and 2.82 pM for LOXL2, Figs. 2E-2G). In this assay, the LOXL2 inhibitor PAT-1251 was used as a control (Figs. 2E-2F). To assess whether LXG6403 is a reversible or irreversible inhibitor, the jump dilution assay was performed. As shown in Fig. 2H, jump dilution of LOXL2 recombinant protein after pre-incubation with increasing doses of LXG6403 showed that there is much less recovery after 120 min jump dilution and at higher LXG6403 doses compared to 30 min dilution and lower concentrations (0% and 14% recovery at 120 min vs. 49% and 37% recovery at 30 min for 30X and 50X ICsos, respectively). This data suggests that LXG6403 is a time- and concentration-dependent irreversible inhibitor. Next, a substrate competition assay was performed and showed that LXG6403 competes with the substrate as evident by the reduction of the inhibitory activity against LOX and LOXL2 at higher doses upon increased substrate concentration (Figs. 21, -2J). To further identify the region of LOXL2 that LXG6403 interacts with, a limited proteolysis analysis was performed. Using this approach, ligand binding sites can potentially be identified through protection of interacting areas from non-specific proteolysis. LC / MS / MS was used to analyze peptides generated from both inhibitor-bound (LXG6403) and -free recombinant LOXL2 protein after limited proteinase K exposure followed by full try ptic digestion (FIG.8). After mapping non-tryptic peptides onto the LOXL2 AlphaFold structure, we identified possible binding regions of LXG6403 that encompass the active site, further confirming the competitive nature of LXG6403 (Fig. 8C).Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0370] After determining the IC50 of lysyl oxidase activity and the inhibition modality of LXG6403. next, characterization of its cellular activity was explored. LXG6403 was tested in a small panel of TNBC cells lines with LOX expression (Fig. 8D) and observed that cellular lysyl oxidase activity was strongly inhibited in all these cell lines with IC50 values < 5 pM (1.3 pM, 1.43 pM, 4.14 pM, and 3.0 pM in MDA-MB-231, HCC143, Hs-578-T, and HCC1937 cells) (Figs.3A-3D). To demonstrate in vivo target engagement, we performed DARTS assay and showed that LXG6403 binds to cellular LOX and protects it from pronase-mediated degradation in a dose-dependent manner (Figs.3E-3G). It also binds to cellular LOXL2, but only at the lower pronase dose (Figs. 3E-3F). On the other hand, there was no protection of LOXL1 by LXG6403 at the pronase and inhibitor doses tested, which altogether supports the specificity for LOX and LOXL2 while no inhibition of LOXL1. Overall, a time- and concentration-dependent irreversible was identified, competitive, bi-thiazole LOX inhibitor with high in vitro potency and good drug-like properties; and therefore, suitable for further investigation in preclinical development studies.Example 3

[0371] To test the effects of bi-thiazole LOX inhibitor LXG6403 on chemoresponse, the TNBC cells, MDA-MB-231 and HCC 1143, were embedded in 3D collagen I and treated them with LXG6403 in combination with different chemotherapy agents that are commonly used in TNBC patients. Treatment with LXG6403 led to enhanced response to the anthracycline, doxorubicin; the platinum-based agent, cisplatin; and the taxane, paclitaxel in both TNBC cell lines (Figs. 3H-3M). Previously it was demonstrated that LOX is responsible for doxorubicin resistance in TNBC. As such, whether the observed effects of LOX family inhibitors on chemotherapy sensitization are specifically due to the inhibition of LOX was explored. To this end, LOX was knocked out via CRISPR-Cas9 (i.e., sgLOX) (Fig. 9A) and showed that this sensitizes cells to chemotherapy agents, doxorubicin, cisplatin, and paclitaxel in collagen I-embedded MDA-MB-231 cells (Figs.9B-9D), similar to the effects that we obtained with our compound, suggesting that LOX inhibition could be a general chemosensitizer in TNBC. Importantly, the reconstitution of LOX with an overexpression plasmid rescued cell \ lability under chemotherapy, further validating the specific role of LOX in chemoresistance.

[0372] Next, the general chemosensitization potential of LXG6403 was tested using organoids derived from the high LOX-expressing TNBC PDX, TM01278, that was chosenPatent Application Attorney Docket No.: USC-838-PCT (1726)based on RNA-Seq data analyses of 15 PDXs in terms of LOX mRNA expression.TM01278 is also resistant to many different chemotherapy agents, including doxorubicin3and docetaxel. LXG6403 was combined with chemotherapeutic agents; doxorubicin, cisplatin, or paclitaxel significantly reduced organoid viability (Fig. 3N) and size (Fig. 30) compared to single agent treatments after 9 days. Finally, the effects of our LOX inhibitor in a dynamic cell migration assay using Real-Time Cell Analyzer (RTCA) were tested, as LOX has been shown to be a key regulator of metastasis. LXG6403 inhibited the migration capacity of the highly migratory MDA-MB-231 cells (Fig. 9E). Overall, these results show that the bi-thiazole LOX inhibitor disclosed herein increases chemoresponse in 3D cultures of TNBC cell lines and PDX organoids and inhibits migration.Example 4

[0373] The primary function of LOX is to crosslink collagen and elastin fibers to organize ECM in the tumor microenvironment (TME). To mimic TME, the human foreskin fibroblast (HFF-l)-derived ECM was utilized and assessed the effects of LXG6403 on canonical functions of LOX in two different experimental settings. In one setting, HFF-1 cells were incubated with LXG6403 during the ECM production (‘ECM from HFF-ls’, Figs. 4A-4B), and in another setting, we seeded HCC1143 cells with or without LXG6403 on top of the HFF-l-derived and decellularized ECM (‘HCC1143 cultured on decellularized ECM’, Figs. 4C-4D). LXG6403 led to reduced deposition of collagen and fibronectin within the ECM, and inhibited their assembly as demonstrated by extracellular collagen and fibronectin staining in either the inhibitor-treated HFF-1 setting (Fig.4A) or in the second setting with TNBC cells treated with the inhibitors and cultured on HFF-l-derived and decellularized ECM (Fig. 4C). It also significantly reduced the amount of insoluble collagen, i.e., crosslinked collagen within the matrix in both settings (Figs. 4B-4D). To test the effects of LXG6403 on drug penetration, we measured doxorubicin autofluorescence in 3D collagen I-embedded TNBC cell lines and PDX organoids under LXG6403 treatment and demonstrated significantly higher doxorubicin penetration upon LOX inhibition in both TNBC cell lines and organoids (Figs. 4E-4G).

[0374] Chemotherapy agents, such as cisplatin or doxorubicin may trigger ROS production and DNA damage. FAK signaling is also known to regulate DNA repair upon DNA damage-inducing chemotherapy. Therefore, the effects of LXG6403 were tested on mitochondrial ROS generation after chemotherapy treatment and observed significantly higher levels of mitochondrial ROS (MITOSOX) upon combination of LXG6403 withPatent Application Attorney Docket No.: USC-838-PCT (1726)cisplatin in 3D collagen I-embedded TNBC cells (Fig 4H). Importantly, more efficient drug penetration and the subsequent increase in chemotherapy -induced ROS led to increased DNA damage, as shown by increased y-H2AX levels (Fig. 41). In addition, reduced FAK phosphorylation was detected (Fig. 41), showing de-activation of FAK / Src signaling, downstream of LXG6403-induced reduction in collagen crosslinking. These ultimately resulted in G1 arrest (shown by p-RB) and apoptosis (shown by increased levels of cleaved Caspase-3 and cleaved PARP) (Fig. 41). Notably, siRNA-mediated LOX knockdown phenocopied the effects of LXG6403 treatment in the observed increase in MITOSOX and change in the downstream signaling when combined with chemotherapy (Fig. 10). These results suggest that the molecular and phenotypic changes upon treatment with LXG6403 are primarily due to on-target effects through specific LOX inhibition. Overall, these data demonstrate that inhibition of LOX with LXG6403, increases drug penetration in 3D culture, induces ROS generation / DNA damage and inhibits FAK signaling, leading to G1 arrest and apoptosis in TNBC cell lines and organoids.Example 5

[0375] To test the potential in vitro toxicity of the bi -thiazole derivatives, the cytotoxicity of parental molecule compound 1 and its derivative LXG6403 (at a dose range of 2.5 pM- 20 pM) was tested on normal cell lines, including human breast cells MCF12A, human umbilical vein endothelial cells HUVEC and HFF-1 cells and observed no significant change in cell viability with LXG6403, while the parental molecule compound 1 had minor cytotoxicity at the highest dose (Fig. 11). Next, the effects of compounds disclosed herein were tested on the activity of monoamine oxidase A and B, which are known to be off-targets of LOX family inhibitors, using a cell-free enzymatic activity assay. Importantly, the parent molecule 1, and lead compound LXG6403, had ICsos of >30 pM for monoamine oxidase A or B (MAO-A / -B).

[0376] The metabolic stability of LXG6403 was tested for its intrinsic clearance rate using both human and mouse liver microsomes and observed good in vitro stability and low / medium clearance. Furthermore, the in vivo toxicity of LXG6403 was tested in a small-scale study based on the PK results. BALB / c mice were treated daily with 25 mg / kg to 75 mg / kg dose range for 5 days with no significant body weight change (Fig. 12A) or blood count change (Figs. 12B-12F) being observed. There was also no apparent organ damage even after 5 days of treatment with 200 mg / kg LXG6403 (Fig. 12G).Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0377] Lastly, the chemosensitization effects of LXG6403 were tested in the resistant TM01278 TNBC PDX model with high LOX expression. Treatment of PDX-bearing mice with LXG6403 given orally in combination with doxorubicin significantly reduced tumor growth and tumor weight compared to LXG6403 and doxorubicin monotherapy groups (Figs. 5A-5B) with no significant change in body weight after 24 days of treatment (Fig.12H). Picrosirius Red Staining (PRS) showed significantly reduced fibrillar collagen content upon LXG6403 treatment (Figs. 5C-5D), and insoluble collagen assay supported PRS results as significantly less insoluble collagen was detected in LXG6403 or combination-treated groups (Fig. 5E). Importantly, MP-SHG showed loss of directionality and organization of the crosslinked collagens in tumor samples treated with LXG6403, further showing the in vivo target engagement of our LOX inhibitor LXG6403 (Fig. 5F, G). Furthermore, combination with LXG6403 increased doxorubicin penetration in vivo (Fig.5H, I). This was followed by increased ROS accumulation (Figs. 5J-5K) and DNA damage (Fig.5L) that further led to FAK inhibition, induction of G1 arrest and apoptosis (Fig.5L), in a similar fashion to the 3D culture results were obtained (Fig.4). Overall, these results show that the bi-thiazole LOX inhibitor LXG6403 overcomes chemotherapy resistance in vivo by reducing fibrillar collagen content / crosslinking, leading to increased drug penetration and ROS accumulation, inhibition of FAK signaling, induction of DNA damage, G1 arrest and apoptosis.Example 6

[0378] The assessment of global changes in fibrillar collagens and to a larger extent e.g., ECM induced upon inhibitor treatment has largely been lacking for the currently available LOX family inhibitors. Therefore, a quantitative pharmacodynamics approach was taken to characterize the global alterations in the ECM composition and organization upon targeting LOX w ith LXG6403 using state-of-the-art ECM-targeted proteomics coupled with mass spectrometry imaging (Fig. 6A). The TNBC PDX tumors collected after 24 days of treatment with LXG6403 alone or in combination with chemotherapy (Fig. 5A) were digested with Collagenase III and the differentially expressed matrix-associated proteins were identified using LC-MS / MS. Intriguingly, the heatmap demonstrated clustering of distinct matrix-associated proteins based on the presence of the LOX inhibitor (Fig.6B). Then, a pathway enrichment analysis was performed using the DAVID pathway analysis tool and showed that more than half of these differentially expressed proteins are significantly enriched in ECM-related processes (Fig. 13). These data suggest that LOXPatent Application Attorney Docket No.: USC-838-PCT (1726)inhibition by LXG6403 has a global biological impact on tumor ECM by rewiring the complex network of matrix- associated proteins.

[0379] To analyze the spatial distribution of the collagen proteins upon targeting LOX with LXG6403, state-of-the-art ECM-targeted mass spectrometry imaging was performed using MALDI FT-ICR. Image segmentation analysis of 2,210 extracted peptides, consisting of majorly the collagen peptides and a smaller fraction of non-collagen ECM proteins showed distinct spatial clustering (Fig.6C) in tumors treated with LXG6403 compared to vehicle or doxorubicin-treated tumors (Fig. 6C), thus suggesting that the proteomic changes exerted by LXG6403 are multilayered, encompassing translational changes in matrix-associated proteins as well as spatial re-organization of fibrillar collagens and also some other ECM proteins. In-depth analysis of the imaging data using MS / MS revealed specific changes in certain post-translational modifications (PTM) within fibrillar collagens, the main targets of LOX, throughout the tumors upon treatment with LXG6403 (Figs. 5D, 9, and 10). Lysine acetylation, lysine oxidation, and proline hydroxylation of fibrillar collagen peptides were mapped and COL3 Al peptide (GPVGPAGK1060SGDRGESGPA), COL1A1 peptide (GFSGLDGAK277GDAGPA), and COL1A2 peptide (GARGEPGNIGFP495) were shown as examples, respectively (Fig. 6E).Based on the imaging data, LXG6403 decreased the lysine oxidation of COL1 Al (middle panel) while lysine acetylation of COL3A1 (upper panel) and proline hydroxylation of COL1 A2 (lower panel) were increased upon LOX inhibition in TNBC PDX tumors (Fig.6E). These results suggest that LOX inhibition by LXG6403 modifies TME not only by altering protein levels / post-translational modifications and the spatial organization of fibrillar collagens, but also changing ECM composition and organization.Table 3. MAO-A / B selectivity of selected derivatives and the LOX activity inhibition _ _ IC50 of 2nd TNBC line, Hs578T _MAO Inhibition IC50 Hs578T ID ID Lysyl Oxidase % Cell A B Activity (IC50, Viability atgM) 10 JLM6403 >30uM >30uM 6403 1.37 100 6512 >30uM >30uM 6512 1 1 100 6609 >30uM >30uM 6609 0.94 92 6619 > 30uM > 30uM 6619 3.63 70 6576 > 30uM > 30uM 6576 2.22 72 6635 >30uM >30uM 6604 0.38 40 6574 >30uM >30uM 6635 1.59 65Patent Application Attorney Docket No.: USC-838-PCT (1726)6616 >30uM >30uM 6613 1.68 70 6592 >30uM ~20uM 6614 0.62 100 6605 >30uM >30uM 6606 1.05 47 6624 > 30uM > 30uM 6616 1.26 100 6561 > 30uM > 30uM 6558 1.27 100 6614 > 30uM > 30uM 6603 1.44 100 6558 > 30uM ~30 uM 6624 1.78 78 6613 > 30uM > 30uM 6574 1.96 90 6569 > 30uM > 30uM 6592 2.27 90 6603 ~15uM > 30uM 6569 4.17 58 6606 ~ 10 uM > 30uM 6561 5.11 100 6604 ~ 10 uM >30uM 6605 5.47 100 6663 > 30uM >30uM6664 > 30uM >30uM6665 ~ 11 uM >30uMExample 7

[0380] The dose-dependent cellular LOX activity inhibition of 78 new LOXi and parental 6403, 6512, and 6487 were first tested in the high LOX-expressing MDA-MB-231 human TNBC cells. Among the 78 new derivatives, 22 showed ICso values <2 pM (Tables 1 and 2 Figs. 2 and 3). Among those, 8 derivatives have <1 pM ICso (6487, 6506, 6512, 6488. 6425, 6532, 6426, 6489, 6539, 6554, 6555, 6606, 6609. 6624, 6603, 6635, 6663, 6678), and 3 of them have ICso ranging from 0.57 - 0.7 pM (6606, 6609, and 6624) (Table 1 and Figs. 15 and 29). The selected 14 derivatives were further validated in another high LOX-expressing human TNBC cell line, Hs578T and correlating ICsos were observed (Fig.29) with some being more active e.g., 6604 (ICso=O.38 pM), suggesting minimal cell linespecific effects (Table 3 and Fig. 29). As a selection filter to exclude potential derivatives with off-targets at an early-stage optimization, we performed MAO-A / B inhibition assay (Fig.30). Thirteen derivatives did not inhibit MAO-A / B (> 30 pM), and 4 of the had MAO-A with 6 < ICso < 15 pM (Table 3). The effect of a few promising analogs were also tested in normal (non-cancerous) endothelial (HUVEC) and breast cells (NMuMG), respectively in terms of cytotoxicity and obtained ICso >70 pM (Fig. 31). Next, recombinant protein-based activity assays were performed for selectivity and on-target activity. Two derivatives (6619 and 6603) were 3-9 times more selective towards LOX than LOXL2 (Table 4, and Fig. 32). Notably, LOXL1 was not inhibited by the majority of the tested LOX inhibitors except 6487, 6576, 6613, and 6619 (Fig. 33).Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0381] Next, recombinant protein-based activity assays were conducted to determine the selectivity of our derivatives and to further confirm on- target activity. A >5-fold selectivity towards LOX over LOXL1 / 2 was initially set as the cut-off to advance optimized candidates. Among the 8 tested LOXi, 3 compounds, 6506, 6512, and 6539, achieved a >5-fold selectivity' towards LOX over LOXL2, while 6532, 6526, and 6487 showed a selectivity ratio between 2 and 3.3 (Table 2 and Fig. 17). Notably, LOXL1 was not inhibited by the majority of the tested LOX family inhibitors except 6487, 6506, and 6425 (to some extent) (Fig. 18). However, considering that 6506 and 6487 fulfilled other important lead optimization criteria, these analogues were included for further testing along with 6512, 6488, 6532, 6539, and 6535 based on their on-target potency, safety, selectivity, and drug-likeness.

[0382] The dose-dependent cellular LOX activity inhibition of 42 new LOXi and parental 6403 were tested in a high LOX-expressing MDA-MB-231 human TNBC cells. Among the 42 new derivatives, potency was less relative to 6403 (>2pM) for 13 derivatives, and essentially equivalent to 6403 for 6 derivatives (1-2 pM). Importantly. 11 novel derivatives had IC50 values <1 pM, including 4 around 500 nM or lower. This included (Table 1 and Fig. 15). Derivatives with the lowest ICsos were further validated in another high LOX-expressing human TNBC cell line, HCC1143. Further, the effect of LOXi was tested on normal (HFF and HEK293T) cell toxicity (Table 1) as anew filter to eliminate derivatives with toxicity towards normal cells. Among 11 potent LOXi, 9 derivatives (6487, 6506, 6425, 6512, 6488, 6426, 6539, 6554, and 6555) minimally reduced normal cell viability' (IC50 of 50-100 pM). While 6532 had variable effects on two different normal cell lines, 6489 negatively affected normal cell viability and was therefore excluded (see Table 1 and Fig. 16). Next, recombinant protein-based activity assays were conducted to determine the selectivity of our derivatives and to further confirm on- target activity7. A >5-fold selectivity towards LOX over LOXL1 / 2 was initially set as the cut-off to advance optimized candidates. Among the 8 tested LOXi, 3 compounds, 6506, 6512, and 6539. achieved a >5-fold selectivity towards LOX over LOXL2, while 6532, 6526. and 6487 showed a selectivity ratio between 2 and 3.3 (Table 2 and Fig. 17). Notably, LOXL1 w as not inhibited by the majority7of the tested LOX family inhibitors except 6487, 6506, and 6425 (to some extent) (Fig. 18). However, considering that 6506 and 6487 fulfilled other important lead optimization criteria, these analogues were included for further testing along with 6512, 6488. 6532, 6539, and 6535 based on their on-target potency, safety, selectivity, and drug-likeness.Patent Application Attorney Docket No.: USC-838-PCT (1726)Table 4: Selected compounds tested in recombinant protein-based activity assays and _ _ cellular binding (DARTS) assays. _Recombinant Protein-Based Assays DARTSID Fold LOX LOXL1 Relative Relative Relative LOX (IC50, LOXL2 activity over (Relative LOX LOXL2 LOXL1 jiM) (ICso, activity at binding binding binding LOXL2gM) 2pM) Relative to untreated control 6403 0.27 0.48 1.78 0.91 5.42 2.66 0.31 6512 0.31 0.54 1.74 1.6 7.9 1.60 0.60 6487 0.22 0.44 2.00 0 5.4 3.90 2.30 6506 0.24 1.22 5.08 0 8.8 2.2 3.3 6425 N / I 1.61 0.56512 0.33 1.99 6.03 1.2 7.9 1.6 0.6 6488 0.12 0.87 1.5 14.1 1.4 0.6 6532 0.1 0.33 3.3 1.16539 02 ~2 10 096426 0.16 0.36 2.256609 0.18 0.11 0.61 1.12 4.89 2.43 1.83 6619 034 1 03 3 03 0 55 441 207 025 6576 0.16 0.13 0.81 0.38 9.96 3.73 1.90 6624 0.09 0.12 1.33 1.86 4.89 1.04 2.80 6614 0.15 0.22 1.47 0.71 4.1 0.49 0.82 6606 1.16 0.59 0.51 0.85 2.62 0.31 0.90 6603 0.21 2 9.52 0.78 7.1 1.76 2.25 6604 0.47 0.13 0.28 1.09 1.55 2.40 1.55 6574 0.48 0.47 0.98 0.81 1.61 1.07 1.61 6592 0.51 1.05 2.06 0.6 7.05 8.17 2.74 6613 0.1 0.28 2.80 0.04 5.07 1.49 0.49 6635 0.21 0.48 2.29 0.87 6.1 3.60 0.33 6616 0.25 0 1 0.40 0.9 1.81 0.62 1.806558 0.57 0.15 0.26 0.7 1.58 1.13 1.58 Note:Recombinant protein-based assay, cell-freeDARTS (Drug Affinity Responsive Target Stability), cell-based

[0383] We tested the binding of 13 selected derivatives to cellular LOX proteins using drug affinity responsive target stability (DARTS) assay (Table 4). We used HA-tagged LOX / LOXLl / LOXL2-overexpressing HEK293T cells with low basal LOX family expression. As shown in Fig. 34, binding to LOX for 4 derivatives (6635, 6603, 6576, and 6592) was stronger than 6403 and was similar for 6 derivates (6606, 6614, 6624, 6609, 6613, and 6619) and less for 4 (6604, 6558, 6574, and 6619). Binding to LOXL2 of 3 derivatives (6635, 6576, and 6592) was stronger than 6403, similar for 3 derivatives (6604, 6619. and 6609) and less for 5 (6603, 6613, 6558. 6574, and 6624) (Fig. 34). As shown in Figs. 19A-19B, protection of LOX was similar for 6487 while it was increased by 1.5 to 2.5- fold for 6506, 6512 and 6488 compared to 6403. Binding to LOXL2 was similar for 6487 and slightly lower for 6506, 6512, and 6488 (Figs. 19A-19C). LOXL1 binding was detected for 6487 and 6506 (Figs. 19A and 19D). Notably, DARTS results were consistent with recombinant protein-based results. In summary, all 4 tested compounds (6506, 6512,Patent Application Attorney Docket No.: USC-838-PCT (1726)6487 and 6488) bind to cellular LOX equally well or better than the parental 6403. The tested compounds also bind to LOXL2 with a variable degree, while LOXL1 binding was detected for 6487 and 6506 at higher doses.Table 5. Functional profiles of selected derivativesFunctional AssaysSynergy' score Synergy scoreSynergy scorein MDA-MB- in MDA-MB- Insoluble ID in TNBC PDOs Doxorubicin231 in 231 in Collagen p-FAK / p- in combination Penetrationcombination combination Relative to SRC / p-H2AX with 30 nM relative to 6403with 1 |1M with 25 nM vehicle doxorubicinDoxorubicin Paclitaxel6403 0.67 0.78 0.67 1 0.53 0.17 / - / 2.7 6512 0.6 04 0.45 1.01 0.48 0.4 / 0.09 / 6 6487 0.34 0.57 0.78 0.926609 0.23 04 0.67 0.97 0.84 0.27 / 0.36 / 1.91 6619 0.39 0.35 0.53 1.23 0.69 0.45 / 0.2 / 1.57 6576 0.26 0.38 0.77 1.17 0.32 0.27 / 0.08 / 2.59 6624 0.21 0.48 0.72 1.22 0.826603 0.34 07 0.59 1.3 0.676592 0.2 0.37 1.07 0.23 / 0.22 / 1 7 6606 0.32 06 1.056614 0.36 0.57 1.19 0.2 / 0.93 / 1.28 6613 0.49 0.73 1.266616 0.9 2.05 1.21 0.06 / 0.44 / 05 6604 0.77 1.616574 0.77 0.866558 0.63 0.556605 1.23 1.236635 1.25 0.716561 082 0866569 0.63 0.556506 0.27 02 0.79 1 Yes 0.54 / 0.13 / 3 5 64256488 0.54 0.45 0.48 0.88 Yes 0.67 / 0.16 / 78 6532 0.81 0.796539 0.6 0.46Patent Application Attorney Docket No.: USC-838-PCT (1726)

[0384] Previously, it was shown that LOX inhibition mediates chemosensitization in TNBCs, but not in ER+ breast cancer. Combining 6 out of 7 selected LOXi (6506, 6512.6487, 6488, 6539, and 6535) with chemotherapy (doxorubicin or paclitaxel) caused synergistic inhibition of cell viability in MDA-MB-231 TNBC cells embedded in collagen I at most of the tested doses (with combination index CKO.8), while ER+ T47D cells were minimally affected (Fig.20). The CI representing synergy’ was lower overall for the new LOXi compared to 6403, indicating better synergy and efficacy (Table 5). Furthermore, the degree of doxorubicin penetration was similar to 6403 for 6 of the shortlisted LOXi (Table 5) Four derivatives (6487, 6488, 6506, and 6512) resulted in synergistic inhibition of viability in TNBC PDX (TM01278) organoids (CI<0.8) (Fig. 21). Finally, reduced collagen crosslinking & fibronectin assembly was observed (Fig. 22), inhibition of FAK / Src signaling and induction of DNA damage (increased p-H2AX) for all these four LOXi (Fig. 23, Table 5), validating on-target inhibition of canonical LOX functions.

[0385] Additionally, we showed that combining 14 or 12 out of 17 selected LOXi with chemotherapy (doxorubicin or paclitaxel, respectively) caused synergistic viability inhibition (CKO.8) in MDA-MB-231 TNBC cells embedded in collagen I at most of the tested doses (Fig. 35 and Table 5). We then tested five derivatives (6576, 6603, 6609, 6619, and 6624) in TNBC PDX (TM01278) organoids, resulting in synergistic inhibition of organoid viability (CKO.8) (Fig.36 and Table 5). These results were further supported by reduction of collagen / fibronectin assembly’ (Fig. 37 and Fig. 39), and inhibition of FAK / Src signaling, induction of DNA damage (increased p-H2AX) (Fig. 38, Table 5), validating on-target inhibition of canonical LOX functions.Table 6. Stability profile of selected derivativesHuman Liver Microsome Clearance Mouse Liver Microsome Clearance ID % Compound Clint % Compound CLint Half-Life Remaining at Half-Life Remaining at30 min pL / min / mg 30 min pL / min / mg minutes 6403 92.14 <58 244.24 61.19 176.61 39.24 6512 98.76 <578 >120 40.5 82.1 84.8 6487 98 <58 >120 81 112 62 6609 9628 87 8056 9646 98 7177 6619 98.87 16.7 415 76.97 76 91.4 6576 10292 1.4 5052.6 93.44 26.8 268.1 6603 92.82 32.7 212.3 73.93 101.2 68.5 6574 875 44.6 160 78.63 79.9 86.7Patent Application Attorney Docket No.: USC-838-PCT (1726)6614 88.45 44.4 156.1 69.32 115.1 60.2 6558 9283 24 1 290 7422 88 788 6616 167 972 1 7.2 56 71.4 97.1 6592 81.14 90.7 76.5 71.38 131.1 53 6624 88.57 39.28 178.16 74.57 97.05 71.59 6635 94.63 20.68 343.84 89.89 188.44 37.45 6506 100.3 <578 >120 53.46 230.4 30.1 6425 10682 <578 >120 79.05 70.9 97.8 6488 81 64 109 78 86 81 6532 80.74 64.6 107.8 62.84 143.9 48.2 " CTRL |45.92 151.85 270.8 25.59 Imipramine" CTRL |27.38 271.27 665.73 10.42 Propranolol" CTRL | 296.99 23.38 267.02 25.97 Terfenadine" CTRL |1503 46.2 355.95 19.5Verapamil*CTRL: Four reference compounds were used as indicators of clearance types involving different metabolic enzymes, CYP1A2, CYP2C19, CYP2D6, CYP3A4.

[0386] To ensure metabolic stability is not compromised for new LOXi while improving on-target potency, we tested the half-life (ti / 2) and intrinsic clearance (CLint) at 2 p M of 11 selected derivatives together with 6403 and 6512 in human and mouse liver microsomes. In mouse liver microsomes, 9 derivatives showed improved stability- compared to 6403, and 4 derivatives out of 9 showed improved stability' compared to 6512. In human liver microsomes, CLint (<57.8), half-life (tl / 2>l 20 min) and % compound remaining (>80%) were excellent, relatively equivalent to parental 6403 for the selected compounds tested (Table 6). clearly showing that our SAR strategy did not compromise metabolic stability. In human liver microsomes, 9 out of 11 derivatives displayed excellent stability compared to 6403 and 6512 (Table 6), clearly showing that our SAR strategy further improved the metabolic stability. To evaluate the intestinal permeability- and potential for efflux-mediated transport, the best 8 derivatives were tested in Caco-2 permeability assay (Table 7). 6619 showed the best permeability profile, followed by 6614, 6574, and 6576 (all better than 6403) whereas 6609 and 6558 showed the highest efflux ratio and poor absorption potential. Plasma protein binding, FBS binding, andPatent Application Attorney Docket No.: USC-838-PCT (1726)plasma half-life of selected derivatives were performed compared to 6403 and 6512 (Table 8).Table 7. Primary drug-like profiles of selected derivatives Cardiotoxicity | ICsoin hERGinhibition Bidirectional Permeability at 10 uM ID MEAN %of MEAN % of ID A-B A-B Percent B-A B-A Percent IC50 (pM) inhibition inhibition Permeability Recovery (%) Permeability Recovery @30pM (10-6 cm / s) (10-6 cm / s) @10JIM (%) ’ 6403 N / C 5.39 12.37 6403 2.2 66 40.5 61 6512 N / C 4.8 14.26 6512 1.2 53 21.2 63 6609 N / C 1.39 6.37 6609 0.5 64 36.8 66 6619 N / C 5.14 18.31 6619 14.8 52 20.7 58 6576 N / C 8.2 12.48 6576 3.39 52 16.5 71 6603 N / C -0.23 0.83 6603 1.5 43 14.3 59 6574 > 30 14.73 34.72 6574 6.5 25 10 37 6614 > 30 20.54 46.42 6614 8.3 35 14.3 51 6558 N / C 8.11 20.87 6558 0.4 56 21.6 52 6592 N / C 3 51 834 6592 076 49 19 1 62 *CTRL |0.47 100 100 “CTRL | 0.1 86 1.5 94 Verapamil Nadolol“CTRL | 22.1 108 18.7 103 Propranolol**CTRL | 6 92 6.2 98Minoxidil* CTRL: Verapamil served as a positive control as IC50 is 0.47 pM.** CTRL: Three reference compounds were used as indicators of different permeability rate; low, high, and moderate in nadolol, propranolol, and minoxidil, respectively.Table 8. Plasma protein binding, FBS binding, and plasma half-life of selected derivativesPlasma ProteinBinding 10% FBS Binding Plasma Half-LifeID% Protein PercentRecov % Protein PercentBound ery' Bound Recovery' at 30min at 120min(» / o)6403 89.11 5486 33.21 46.84 94.47% 92.24% 6512 89.73 4441 45.49 52.05 94.53% 69.79% 6576 91.4 5604 41.63 26.52 97.11% 88.40% 6609 96.26 31 91 62.12 11.24 95.08% 75.72%Patent Application Attorney Docket No.: USC-838-PCT (1726)6619 91.24 3745 56.02 97.19 89.39% 64.67%

[0387] All 10 derivatives tested did not inhibit hERG at 10 pM (Table 7), indicating no cardiotoxicity risk. To test potential off-targets at a larger scale, we performed kinome profiling (ScanMAX Kinase Panel, KINOMEscan, Eurofins Discovery) and CEREP Screen (SafetyScreen47 Panel. Eurofins Discovery) with select derivatives (6487, 6506, 6576 and 6609) along with two initial leads 6403 and 6512. In general, excellent selectivity scores (0.002 and 0.01 for 6487 and 6506, respectively with %Control<10) were obtained (Table 9). Considering ICso values for recombinant LOX inhibition (0.2-0.3 pM) is much lower than 10 pM used in the kinome profiling, it was concluded that the LOXi have no major kinase off-targets. In CEREP assay testing off-target inhibition of 78 proteins (ion channel, receptors, and enzymes), there were only a few inhibited by the LOXi (Table 9).Recombinant COX1 / 2 were inhibited at an RC50 (similar to ICso) range of 0.5-1 pM (still 2.5 to 5 times higher than recombinant LOX IC50) by 6403, 6506, and 6487. The other potential off-targets were inhibited at an RC50 value of 2.7 - 9.56 pM, much higher than LOX inhibition (0.2 - 0.3 pM), except OPRD1 which was inhibited at 0.32 pM by 6506. Notably, MAO-A was inhibited at an RC50 value of 6.95 only by 6487. Further, potential MAO inhibition was tested in a second assay with 6 of the selected compounds (6506, 6512. 6487, 6488, 6539, and 6532) since MAO-A / B are known potential off-target enzymes affected by LOX family inhibitors. While MAO-A / B ICsos were >30pM and equivalent to 6403 for 6512, 6506, 6532, and 6539, MAO-A ICso was lower for 6487 and relatively lower for 6488, indicating a lower selectivity for these two inhibitors (Table 9 and Fig. 24). Additionally, we obtained high selectivity, especially for promising lead derivative, 6576 (Table 9 and Fig.39). The 6576 reverse amide group leads to removal of HTR2B and COXI inhibition compared to 6403 in the safety screen and excellent selectivity7with only COX2 being inhibited. Comparing the results for 6403 and 6512, the meta acetamido group gives less selectivity to 6512 than the para in the kinome panel; however, it improves selectivity towards COX1. Comparing the results for 6403 and 6609 -the isoindolin-l-one group of 6609, leads to poor selectivity' compared to theacetamidophenyl.Patent Application Attorney Docket No.: USC-838-PCT (1726)Table 9. Selected compounds tested in kinome profiling, CEREP assay and MAO activity to assess potential off-targetsg 1 g i i I g 1 1 g g 1 1 g2O § I g g I g g g 1 § I g12i7 22o 2u O Q o 2u o u25Q 1 g§mg 11g1 §igjQcO' t? c O-'l£ 1 \© § i- >08O' ro 2 - sPatent Application Attorney Docket No.: USC-838-PCT (1726)

[0388] To test potential off-targets, kinome profiling (ScanMAX Kinase Panel, KINOMEscan. Eurofins Discovery ) and CEREP Screen (SafetyScreen47 Panel, Eurofins Discovery) was performed with two lead LOXi (6487 and 6506) along with the parental 6403.

[0389] The effects of LXG6403 were validated in vivo regarding the ability of suppressing tumor growth, metastasis, and potentiating the efficacy of chemotherapy or immunotherapy. Initially, the chemosensitization effects of LXG6403 were tested in the relatively cyclophosphamide (CP) resistant TM00099 TNBC PDX model with high LOX expression. Treatment of PDX-bearing mice with LXG6403 given orally in combination with CP significantly reduced tumor growth and tumor weight compared to LXG6403 and doxorubicin monotherapy groups (Fig. 25A). It was observed that LXG6403 was able to reduce lysyl oxidase activity in vivo in an in vivo pharmacodynamic experiment (Fig. 25B).Furthermore, LXG6403 or its combination with chemotherapy showed excellent safety7profiles as evidenced by observed no significant changes in body weight (Fig. 25C) and major blood counts (Figs. 25D and 25E). Second, the effects of LXG6403 on suppressing spontaneous metastasis of highly metastatic mammary tumor model, MMTV-PyMT, were tested (Fig. 26A). As the changes in circulating tumor cells (CTC) are an indicator of metastatic dissemination, the number of CTCs in blood was determined and it was observed that they were significantly reduced upon LXG6403 (Fig. 26B) without showing any significant body weight change (Fig. 26C). Notably, although the tumor weight was not changed by LXG6403 alone (Fig. 26D), in the percentage of lung nodules was significantly reduced by LXG6403 (Figs. 26E and 26F). Next, the effects of LXG6403 on overcoming immunotherapy resistance were tested (Fig. 26). Using the same transgenic model (MMTV-PyMT model), tumor transplantations were performed and treated mice with LXG6403 for a week and then treated with anti-PD-Ll antibody (Fig. 27A). The combination of LXG6403 and anti-PDLl was shown to effectively suppressed tumor growth (Fig. 27B) without any major toxicity7(Figs.27C - 27G). These data support that LXG6403 overcomes resistance to both standard-of-care chemotherapies and immunotherapies in breast cancer. In addition to that, as a single agent, LXG6403 is effective on the suppression of spontaneous metastasis. Finally, the effects of LXG6403 on tumor growth in kidney cancer in vivo were tested because fibrosis plays an important role in the development and progression of renal cell carcinoma similar to TNBC. In this case, LXG6403 was combined with one of the current standard-of-care therapy, Axitinib, in kidney cancer xenografts 786-0. Notably, combination of LXG6403 and Axitinib lead toPatent Application Attorney Docket No.: USC-838-PCT (1726)s Cd #iompoungnificant tumor regression in half of the mice while other half had stabilized tumors compared to single agent treatment (Fig. 28). These data implicate that LOX familyCd IDompouninhibitors disclosed herein provide the potential of extended indications regarding fibrosis- Cd #ompoun related tumors, e.g. TNBC, kidney cancer, and pancreatic cancer, etc.CdompounTable 10: Protein Binding Assay IDPlasma ProteinBinding 10% FBS Binding Plasma Half-Life % Percent % Percent at Protein Recovery Protein at 30minBound B Recovery 120minound (%) ‘6403 89.11 54.86 33.21 46.84 94.47% 92.24%Cd IDompoun 1 6512 89.73 44.41 45.49 52.05 94.53% 69.79%2 6576 91.4 56.04 41.63 26.52 97.11% 88.40%3 6609 96.26 31.91 62.12 11.24 95.08% 75.72%4 6619 91 24 3745 5602 97 19 8939% 6467%Table 11Cardiotoxicity | IC50 in hERG Bidirectional Permeability at 10 uM inhibitionMEAN % MEAN % A-B B-A of of A-B Percent B-A Percent ICso Permeability ( Permeability ( inhibition inhibition Recovery(%) Recovery(%) 10-6 cm / s) 10-6 cm / s) @10uM @30uM6403 N / C 5.39 12.37 6403 2.2 66 40.5 61 1 6512 N / C 4.8 14.26 6512 1.2 53 21.2 63 2 6609 N / C 1.39 6.37 6609 0.5 64 36.8 66 3 6619 N / C 5.14 18.31 6619 14.8 52 20.7 58 4 6576 N / C 8.2 12.48 6576 3.39 52 16.5 71 5 6603 N / C -0.23 0.83 6603 1.5 43 14.3 59 6 6574 > 30 14.73 34.72 6574 6.5 25 10 37 7 6614 > 30 20.54 46.42 6614 8.3 35 14.3 51 8 6558 N / C 8.11 20.87 6558 0.4 56 21.6 52 9 6592 N / C 3.51 8.34 6592 0.76 49 19.1 62

[0390] These and other modifications and variations to the present invention may be practiced by those of ordinary skill in the art, without departing from the spirit and scope of the present invention, which is more particularly set forth in the appended claims. In addition, it should be understood that aspects of the various embodiments may be interchanged both in whole or in part. Furthermore, those of ordinary skill in the art will appreciate that the foregoing description is by way of example only and is not intended to limit the invention so further described in such appended claims.

Claims

Patent Application Attorney Docket No.: USC-838-PCT (1726) What Is Claimed;1. A compound of Formula (A):or a pharmaceutically acceptable salt thereof, wherein:Xis CH orN;R1is Ci-8 alkyl or Ci-8 haloalkyl:R2and R3are independently hydrogen, Ci-8 alkyl, Ci-8 alkylamino, -CH2-phenyL or -C(O)Ra; wherein each Rais independently Ci-8 alkyl or phenyl; R4, R5, R6, and R7are each independently hydrogen, halogen, Ci-8 alkyl, Ci-8 alkylamino. Ci-8 alkoxy, -ORb. -N(Rb)2, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2RC. -S(O)2N(Rb)2, or -NRbS(O)2Rc,or R4and R5or R6and R7combine with the intervening atoms to form a phenyl, 5 to 6-membered heteroaryl, or 5 to 6-membered heterocycloalkyl which is saturated or partially unsaturated, wherein the phenyl, heteroaryl and heterocycloalkyl are optionally substituted with one or more R8; each R8is independently halogen, Ci-8 alkyl, -ORb, oxo (=0), -N(Rh)2, -C(O)ORb, - C(O)RC, -C(0)N(Rb)2, or -NRbC(0)Rc;each Rbis independently hydrogen, Ci-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; or two Rbcombine with the nitrogen atom to which they are attached to forms a 4 to 8-membered heterocycloalkyl which is optionally substituted with one or more Rd;each Rcis independently Ci-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; andeach Rdis independently halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, wherein the phenyl orPatent Application Attorney Docket No.: USC-838-PCT (1726)heteroaryl is optionally substituted with one or more halogen, -OH, -OCH3, -NH2. -NHCH3. -N(CH3), -CH3, -C(O)OH. -C(O)OCH3. or -NHC(O)CH3.R5Rt ±. R6V'^X'^R2. The compound of claim 1, wherein when ring Ais '7, then at least one of R4, R5, R6. or R7is not hydrogen.

3. The compound of claim 1, wherein the compound is of Formula (1-4):or a pharmaceutically acceptable salt thereof, wherein:Xis CH orN;R1is -CH3, -CH2CH3, or -CF3;R2and R3are independently hydrogen, C1-8 alkyl, or -C(O)Ra; wherein each Rais independently C1-8 alkyl or phenyl;R4, R5, R6, and R7are each independently hydrogen, halogen, C1-8 alkyl, C1-8 alkylamino. C1-8 alkoxy, -ORb. -N(Rh)2, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2RC. -S(O)2N(Rb)2, or -NRbS(O)2Rc,or R4and R5or R6and R7combine with the intervening atoms to form a phenyl, 5 to 6-membered heteroaryl, or 5 to 6-membered heterocycloalkyl which is saturated or partially unsaturated, wherein the phenyl, heteroaryl, and heterocycloalkyl is optionally’ substituted with one or more R8, each R8is independently halogen, C1-8 alkyl, -ORb, oxo (=0), -N(Rb)2, -C(O)ORb, - C(O)RC, -C(0)N(Rb)2, or -NRbC(0)Rc;each Rbis independently hydrogen, C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; or two Rbcombine with the nitrogen atom to which they are attached to forms a 4 to 8-membered heterocycloalkyl which is optionally substituted with one or more Rd;each Rcis independently C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; andPatent Application Attorney Docket No.: USC-838-PCT (1726)each Rdis independently halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, wherein the phenyl or heteroaryl is optionally substituted with one or more halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), -CH3, -C(O)OH, -C(O)OCH3, or -NHC(O)CH3;R5Rt Jx^R6wherein when ring Ais ', then at least one of R4, R5, R6, or R7is not hydrogen.

4. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein X is N.

5. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein X is CH.

6. The compound of any one of claims 3-5, or a pharmaceutically acceptable salt thereof, wherein R5is hydrogen.

7. The compound of any one of claims 3-6, or a pharmaceutically acceptable salt thereof, wherein R4is hydrogen, -C(O)ORb, -C(O)RC, or -NRbC(O)Rc.

8. The compound of any one of claims 3-7, or a pharmaceutically acceptable salt thereof, wherein R6and R7are each independently hydrogen, halogen, C1-8 alkylamino, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc. -S(O)2RC. - S(O)2N(Rb)2, or -NRhS(O)2Rc; wherein one of R6or R7is not hydrogen.

9. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein Xis N;R4is hydrogen;R3is hydrogen;R6is hydrogen, halogen, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc; and R7is halogen, C1-8 alkylamino, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, - S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc.

10. The compound of claim 9, or a pharmaceutically acceptable salt thereof, wherein R6is hydrogen or halogen.

11. The compound of claim 9 or 10, or a pharmaceutically acceptable salt thereof, wherein R7is -C(O)OH, -NHC(O)CH3, -NHC(O)CH2CH2OCH3, -C(O)NH-(CH2)4- phenyl. -C(O)NH-phenyl, -C(O)NH-(2,4-difluoro-phenyl), or -C(O)NH-phenyl-4- acetamide.Patent Application Attorney Docket No.: USC-838-PCT (1726)12. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein X is CH;R4is hydrogen;R ' is hydrogen;R6is hy drogen, halogen, Ci-s alkylamino, -C(O)ORb, -C(O)RC, -NRbC(O)Rc, - S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc; andR7is hydrogen, halogen, Ci-s alkylamino, -C(O)ORb, -C(O)RC, -NRbC(O)Rc, - S(O)2RC, -S(O)2N(Rb)2, or -NRbS(O)2Rc;wherein one of R6or R7is not hydrogen.

13. The compound of claim 12, or a pharmaceutically acceptable salt thereof, wherein R6is hydrogen or halogen; andR7is -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, -NRbC(O)Rc, -S(O)2RC, -S(O)2N(Rb)2, or - NRbS(O)2Rc.

14. The compound of claim 12 or 13, or a pharmaceutically acceptable salt thereof, wherein R7is -CH2NH2, -C(O)OH, -C(O)NHCH3. -C(O)N(CH3)2, -NHC(O)CH3, - NHC(O)CH2CH2OCH3, -C(O)NH-(CH2)4-phenyl, -C(O)NH-phenyl, -C(O)NH- (2,4-difluoro-phenyl), -C(O)NH-phenyl-4-acetamide, -NHC(O)-CH2-(4-fluoro- phenyl), -NHC(O)-CH2-(4-cholo-phenyl), -NHC(O)-(4-methoxy-phenyl), - NHC(O)-3-pyridynyl, or -NHSO2CH3.

15. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein R4and R ' combine together with the intervening atoms to form a 5 to 6-membered heteroaryl having 1, 2, or 3 heteroatoms selected fromN, O, and S, or a 5 to 6- membered heterocycloalkyl with is saturated or partially unsaturated, having 1 or 2 heteroatoms selected from N and O, wherein the heteroaryl or heterocycloalkyl is optionally substituted with one or more R8.

16. The compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein R4and R5combine with the intervening atoms to form a pyrrole, imidazole, triazole, thiazole, oxazole, pyridine, furan, pyrrolidine, pyrrolidinone, irmdazolidinone, tetrahydropyran, or piperidinone.

17. The compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein R4and R5combine with the intervening atoms to form a pyrrolidine, pyrrolidinone, or imidazolidinone.

18. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein R6and R7combine together with the intervening atoms to form a 5 to 6-memberedPatent Application Attorney Docket No.: USC-838-PCT (1726)heteroaryl having 1, 2, or 3 heteroatoms selected fromN, O, and S, or a 5 to 6- membered heterocycloalkyl with is saturated or partially unsaturated, having 1 or 2 heteroatoms selected from N and O, wherein the heteroar l or heterocycloalkyl is optionally substituted with one or more R8.

19. The compound of claim 18, or a pharmaceutically acceptable salt thereof, wherein R3and R6combine together with the intervening atoms to form a pyrrole, imidazole, triazole, thiazole, oxazole, pyridine, furan, pyrrolidine, pyrrolidinone, imidazolidinone, tetrahydropyran, or piperidinone.

20. The compound of claim 18, or a pharmaceutically acceptable salt thereof, wherein R4and R5combine together with the intervening atoms to form a pyrrolidine, pyrrolidinone, or imidazolidinone.

21. The compound of claim 1, wherein the compound is of Formula (1-3):or a pharmaceutically acceptable salt thereof, wherein:R1is -CH3, -CH2CH3, or -CF3;R2and R?are independently hydrogen, Ci-s alkyl, or -C(O)Ra; wherein each Rais independently C1-8 alkyl or phenyl;R4is hydrogen. Ci-s alkyl, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc; R5is hydrogen, Ci-s alkyl, -C(O)ORb, -C(O)RC, -C(O)N(Rb)2, or -NRbC(O)Rc; each Rbis independently hydrogen, Ci-s alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; or two Rbcombine together with the nitrogen atom to which they are attached to forms a 4 to 8-membered heterocycloalkyl which is optionally substituted with one or more Rd;each Rcis independently C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, each of which is optionally substituted with one or more Rd; andeach Rdis independently halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CHs), C1-8 alkyl, phenyl, or 5 to 6-membered heteroaryl, wherein the pheny l or heteroaryl is optionally substituted with one or more halogen, -OH, -OCH3, -NH2, -NHCH3, -N(CH3), -CH3, -C(O)OH, -C(O)OCH3, or -NHC(O)CH3.

22. The compound of claim 21, or a pharmaceutically acceptable salt thereof, whereinPatent Application Attorney Docket No.: USC-838-PCT (1726)R4is hydrogen, -NHC(O)CHs, or -C(O)OCHsCHs; andR5is hydrogen. -NHC(O)CHs, -NHC(O)-Ph-NHC(O)CHs, -NHC(O)-Ph-2,4- difluoro, -C(O)OH, -C(O)NHPh, or -C(O)NH-Ph-NHC(O)CH3.

23. The compound of claim 22, or a pharmaceutically acceptable salt thereof, wherein R4is hydrogen.

24. The compound of claim 22, or a pharmaceutically acceptable salt thereof, wherein R’ is hydrogen.

25. The compound of any one of claims 1-24, or a pharmaceutically acceptable salt thereof, wherein R1is -CHs.

26. The compound of any one of claims claim 1-25, or a pharmaceutically acceptable salt thereof, wherein R2is hydrogen, -CHs. -CH2CH3, or -C(O)CHs.

27. The compound of claim 26, or a pharmaceutically acceptable salt thereof, wherein R2is hydrogen.

28. The compound of any one of claims 1-26, or a pharmaceutically acceptable salt thereof, wherein R3is hydrogen or -CHs.

29. A compound selected from:ID Structure6415 Jp)H2N |6425 A?II / ' / L / H2N H H6426 N-L / H tn / -s' N N NH2N H H 1 / H HN" A6487\ Y nH2N s A-S 06488 Pl pppH2N HH6489 A-c; l\rxNX^ / J"'NH2N HozPatent Application Attorney Docket No.: USC-838-PCT (1726)6505 N-4 -OzZ-N H H6506 / H Q H:6512 ZI <H2N A. S, V-SnH ifA L z ° 0k zIZ6519-A / Z -W^-o - cZ zf z= A^ 2=.IM CI\ O^6519-B o \\ A A- / -c N (D~i JCQAbZI I VH(A G "a6522 ZIx °6523 N / r f vl " A AAs' ' NANA7 °H2N H6524A xA ra,H2N H6528nA iA YWH2N H0 F1 H6529 NA -1 I (I T n ]| ]ii / \ AL A o XAsH2N H1 A JO6530 N-V / l jf j HAsH2N HPatent Application Attorney Docket No.: USC-838-PCT (1726)06531H2N As' H / H / SKN IIA / J ' -U | M > N M-Y6532 H2N As H \ / Sy^NH2NH6533As' ' N^N^AA0H2N HH2N6538 ys NH26539H2N A H o1''0 65542A AH N HH6555 - N< vA / .- A A rr J -HH2N As1^Ho6556H2N H6558nAu rr"^H2N As' '' H0Patent Application Attorney Docket No.: USC-838-PCT (1726)0 A" N / 2 JL ji6559. AN y-X \ I || HAsH2N H06560KA. A "H2N INYA A u6561 H2N AsNN H N YX oh[I I KH6562H2II / Xk JY A A AN AsNN H NhAH6563 N^AX xi ZVAH2ANH MNH NH26568H2N As^^ N^N H^^A-06569 NX__A^H2NAs^^ N^N H^^AHNr^°N'<Lxn6570H2NAs' " N^NH^^A HN^0.u6571 AS^ VS^^A0H2N HHN^ „AHPatent Application Attorney Docket No.: USC-838-PCT (1726)65726573 > _ O2As^ ^ NAN^H N HN / ) Gz—\ _ / I / ZI6574 N-VO f I *]T H2N As k Zro HZ / 6574-B N As— e i r IT 1As AY —HH2N A Z=z H III " Z \- J AA6574-C NA— A I | II 1HH2N 17X Y IIz \\s-x / , / S^NH2O HN A N ' Y-N no=\6574-D ZI / A TASHL A J6574-E6575Z^-S N N SH2N H / H96576 S 'V / NYNYN / N / H M2 NN'ASZY x—- " SO L JH1 H [ ||6577N A-Al \ T TfY »*w 0H2N HPatent Application Attorney Docket No.: USC-838-PCT (1726)i A JOL6578 Vfl jTj HFH2N As7HT6579 oo4 / ) z oZ / \\ e XLNTZ6580 Jo M AOHH2NVAA z / A S\\ / N\6581,, z<xl cvZ=IkT!A Ar6582... A rA A65916592 A XX XV-XsH H NH26594 NA _ / A JTT TH2N HJ / ~s ^VN»,6595 o A^X XJOAN HAS nAH2NK OH H6603NX ^-s XiX0\ V-HKoH H6604YA XTPatent Application Attorney Docket No.: USC-838-PCT (1726)6605JX / QYJ H O6606 N-\H2N jL sy v-s j T IInh / H H6607 I S V-S r ¥ P ¥ 0HO6609 N¥ _ ¥X jf ll¥NHAY N^N^^""7H2N H / H HN-46613H2N JIL. s / A V-NH i^ ¥ 06614Nn X-xi XX XH2N H H1 H6616NX-¥I ¥¥N¥ / -s ' N¥N-¥¥~C|0H2N H / 1 H6619H2N s Vs 0s, XX XV¥ o6621 0 N N NN^\ IIII L I H H N-^X^-xH 'QQ HKLH H6622° Y-VT XX V^06624 N. y-¥3 jXXnh / —-S NHHPatent Application Attorney Docket No.: USC-838-PCT (1726)O6625xH -sZ pH2N H 'OoX JL "6626 / — g NNHH OHH2NxH H6628 A I U IFTFo6631H2N H QoH2N. _S #~-S6634 Y MA JL L / NJNH\ HA~FF F J H H6635 AN^NMM"NH2N A s / — Vs / T II )=o / H / >7~~S 'r'" N.6636 N v-< O JL L >°H2N As H6706 N A- / J1 JL L >°H2N As HX HV-Nx„ H H o6637 V • •S.\A Il,o s.nM °H2K O H p6638NL / ~A Q L L zN—Patent Application Attorney Docket No.: USC-838-PCT (1726)O6639H2NYH Xk C / 6641 s WIZ°Z'zx0 Ok=6657 XZ / ^—S N NH2N H HTZ xzz6658 J zzZ^=M C Z Z —.Xs W - I}z= k z6663 N < zXz <>^z= 0z2X6664H NYW~X MjQrVN-7 N6677 jOCcli / ~~S N N NH2N | H6678H2NAB'S4>__X / NVS ^YNY^N H / 6681H2N'J" S'^X-S kJLHH2NK.. vlH6670 Ws< TVYs Y kYJ ^o6679H z~-sNNH2N |Patent Application Attorney Docket No.: USC-838-PCT (1726)6680 J I?NII" A A _ / if Nu H2N 0 — 0 Q L UH6681 -A!'Y / NYNYN / N / 2I X H NA' VS zHJ I - N6681 -BH2N Ar Vsz — y zHz y?A —<- 6698 z^ Z=6699 Z z / H06700(Jl H06701 M NA / ~~W XSII J HH2N^ / ^SN30. The compound of any one of claims 1-29, or a pharmaceutically acceptable salt thereof, wherein the compound, or pharmaceutically acceptable salt thereof, is a lysyl oxidase (LOX) inhibitor.

31. The compound of claim 30, or a pharmaceutically acceptable salt thereof, wherein the compound, or a pharmaceutically acceptable salt thereof, inhibits cell-based lysyl oxidase activity and / or recombinant protein based lysyl oxidase activity assays.

32. The compound of claim 30 or 31, wherein the compound, or pharmaceutically acceptable salt thereof, is selective for LOX.

33. A pharmaceutically composition comprising a compound of any one of claims 1-29, or a pharmaceutically acceptable salt thereof.

34. A method of treating a disease of a condition associated with the modulation or inhibition of lysyl oxidase (LOX), the method comprising administering to a subject in need thereof, a therapeutically effective amount of a compound of any one ofPatent Application Attorney Docket No.: USC-838-PCT (1726)claims 1-29, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 32.

35. The method of claim 34, wherein the therapeutically effective amount of the compound or its pharmaceutically acceptable salt is administered to treat a neurodegenerative disease, an angiogenesis-related disease, Alzheimer's disease, fibrosis including: liver fibrosis, pulmonary fibrosis, renal fibrosis, myocardial fibrosis, cystic fibrosis, idiopathic pulmonary fibrosis, radiation-induced fibrosis, ocular fibrosis, Peylony's disease and scleroderma, or respiratory disorders, abnormal wound healing and repair, postoperative surgery, cardiac arrest-related fibrosis, excess or abnormal deposition of fibrotic material, all associated with disorders such as Crohn's disease and inflammatory bowel disease, liver, kidney and pancreas fibrosis, diabetes, cerebral hemorrhage with amyloidosis, cardiac hypertrophy, Hutchinson-Gilford Progeria Syndrome, retinopathy, chemoresistance, and / or a kidney disorder including: kidney fibrosis, renal fibrosis, acute kidney¬ injury, chronic kidney disease, diabetic nephropathy, glomerulosclerosis, vesicoureteral reflux, hypertrophic scarring, keloids, diabetic skin, tubulointerstitial renal fibrosis and / or glomerulonephritis.

36. The method of claim 35, wherein the cancer is selected from the group comprising lung cancer; breast cancer; colorectal cancer; anal cancer; pancreatic cancer; prostate cancer; ovarian carcinoma; liver and bile duct carcinoma; esophageal carcinoma; non-Hodgkin's lymphoma; bladder carcinoma; carcinoma of the uterus; glioma, glioblastoma, medullablastoma, and other tumors of the brain; myelofibrosis, kidney cancer; cancer of the head and neck; cancer of the stomach; multiple myeloma; testicular cancer; germ cell tumor; neuroendocrine tumor; cervical cancer; oral cancer, carcinoids of the gastrointestinal tract, breast, and other organs; signet ring cell carcinoma; mesenchymal tumors including sarcomas, fibrosarcomas, haemangioma, angiomatosis, haemangiopericytoma, pseudoangiomatous stromal hyperplasia, myofibroblastoma, fibromatosis, inflammatory myofibroblastic tumour, lipoma, angiolipoma, granular cell tumour, neurofibroma, schwannoma, angiosarcoma, liposarcoma, rhabdomyosarcoma, osteosarcoma, leiomyoma and / or a leiomysarcoma.

37. The method of claim 36, wherein the breast cancer is triple negative breast cancer (TNBC), estrogen receptor-positive breast cancer, metastatic breast cancer, HER2 positive breast cancer, or a combination thereof.Patent Application Attorney Docket No.: USC-838-PCT (1726)38. The method of claim 34, wherein the therapeutically effective amount of the compound or its pharmaceutically acceptable salt is administered to reduce cardiotoxicity side effects of chemotherapy and / or immunotherapy.

39. The method of any one of claims 34-38, wherein the therapeutically effective amount of the compound or its pharmaceutically acceptable salt is administered as a single agent or in combination with chemotherapy, immunotherapy and / or radiotherapy in both adjuvant and neo-adjuvant settings.

40. The method of any one of claims 34-38, wherein the therapeutically effective amount of the compound or its pharmaceutically acceptable salt is administered with a second therapeutic agent selected from an anti-cancer agent, an antiinflammatory agent, an anti-hypertensive agent, an anti-fibrotic agent, an anti- angiogenic agent, and / or an immunosuppressive agent.

41. The method of any one of claims 34-40, wherein the subject has failed at least one anti-cancer therapy.

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