Diazaquinolines and related compounds as antiviral agents

Abstract

Disclosed herein are substituted isomeric naphthyridinyl benzamides and related compounds as broad-spectrum GHP-class polymerase inhibitors, and pharmaceutical compositions thereof, utilized for the treatment and prevention of viral infections. The inventive compounds may be used to treat a paramyxovirus infection, an orthoparamyxovirus infection, a respirovirus infection, a morbillivirus infection, or a henipavirus infection.

Description

Attorney Docket No. 10013-110W01DIAZAQUINOLINES AND RELATED COMPOUNDS AS ANTIVIRAL AGENTS CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to United States Provisional Patent Application No.63 / 737,231 filed December 20, 2024, and United States Provisional Patent Application No. 63 / 747,637 filed January 21, 2025, the disclosures of which are incorporated herein by reference in their entireties.STATEMENT ACKNOWLEDGING GOVERNMENT SUPPORT

[0002] This invention was made with government support under Grant Nos. U19AI171403 and AI071002 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND

[0003] There remains a need for improved compositions and methods for treating or preventing viral infections, such as paramyxovirus infections, including orthoparamyxovirus infections.BRIEF DESCRIPTION OF THE FIGURES

[0004] FIGs. 1A-1F depict that GHP-88310 prophylaxis in contact animals blocks direct contact transmission of CDV. (FIG. 1A) Overview of the prophylactic assessment of twice-daily GHP-88309 or GHP-88310 to prevent direct contact transmission of CDV in ferrets. Source ferrets were intranasally infected with 2xl05TCIDso units of recombinant CDV. Contact ferrets received oral treatment, beginning 12 hours prior to co-housing with infected source animals. (FIG. IB) Survival curves with statistical analysis by log-rank (Mantel-Cox) test and indicated median survival (m.s.). (FIG. 1C) PBMC associated viremia and (FIG. ID) nasal lavage titers, quantified byTCIDSo. (FIG. IE) Lymphocyte counts; normal reference range indicated by shading. (FIG. IF) Neutralizing antibody titers. (FIGs. 1C-1F) Symbols represent individual animals; lines represent the (FIGs. 1C-1E) geometric mean or (FIG. IF) mean. (FIGs.1C-1E) Two-way ANOVA followed by either Fisher's LSD (FIG. 1C), Dunnet's post hoc multiple comparisons test (FIG. ID) or Tukey's multiple comparisons test (FIG. 1G). (FIGs. 1C-1D) Period of cohousing indicated by shading, lod, limit of detection.

[0005] FIGs. 2A-2E depict CDV transmission dynamics in a ferret model. (FIG. 2A) Schematic of airborne CDV transmission setup. Contact ferrets were co-housed with CDV-infected source ferrets in either an open- or closed-cage system from study days 5 to 11 to allow airborne exposure. (FIG. 2B) Survival curves with statistical analysis by log-rank (Mantel-Cox) test and indicated median survival (m.s.). (FIG.2C) PBMC associate viremia titers and (FIG. 2D) nasal lavage titers. (FIGs. 2C-2D) Data represented as geometric mean with geometric SD except open cage air transmission group: symbols representAttorney Docket No. 10013-110W01individual animals to show variability, with lines representing geometric means. (FIG. 2E) Schematic comparing clinical signs and disease progression of CDV infection in ferrets following intranasal inoculation versus airborne contact exposure, aligned to days post infection (intranasal) or exposure (open / close cage air transmission). Light colored block indicates disease onset of the specified clinical sign of the first ferret of the group. Dark colored block shows when all ferrets of the group reached the specified disease stage.

[0006] FIGs. 3A-3G depict that airborne transmission of CDV is prevented or controlled by both prophylactic and therapeutic GHP-88310 treatment of contact animals. (FIG. 3A) Study schematic of once- or twice-daily prophylactic and therapeutic treatment of contact animals to prevent infection via airborne transmission of CDV. Source ferrets were intranasally infected with 2x10sTCID50 units of recombinant CDV. Five days post-infection, they were co-housed with uninfected contact ferrets in a close-caged system for six days, after which all animals were individually housed for the remainder of the study. Treatments were initiated at various time points relative to the start of co-housing as indicated. On day 31 of the study, all treatment groups were challenged intranasally with 2xl05TCID50 units of recombinant CDV. Initial challenge is denoted by dark shading, re-challenge by light shading. (B) Survival curves from initial challenge. (FIG. 3C) Lymphocyte counts from initial challenge; normal reference range indicated by shading. Viral titers in (FIG. 3D) nasal lavages, (FIG. 3E) PBMCs and (FIG. 3F) neutralizing antibody titers measured throughout both the initial and re-challenge. (FIG. 3G) Survival curves following re-challenge. (FIGs. 3B and 3G) Log-rank (Mantel-Cox) test with median survival (m.s.) shown. (FIGs. 3C-3F) Symbols represent individual animals; lines represent the (FIGs. 3C-3E) geometric mean or (FIG. 3F) mean. (FIGs. 3C-3E) Two-way ANOVA followed by Dunnet's post hoc multiple comparisons test, comparisons are to the vehicle group, dpc, days post co-housing. lod, limit of detection.

[0007] FIGs. 4A-4F depict that therapeutic GHP-88310 treatment in CDV-infected ferrets blocks airborne transmission to untreated contacts. (FIG. 4A) Schematic overview of GHP-88310 treatment in infected source ferrets to assess its ability to prevent airborne transmission to untreated contacts. Source ferrets were intranasally infected with a lower dose of 2xl03TCID50 units of recombinant CDV. Treatment was administered twice daily, beginning four days post-infection. Infected source ferrets were then co-housed with untreated contact ferrets for five days. Quantification of virus in (FIG. 4B) nasal lavages and (FIG. 4C) PBMCs. (FIG. 4D) Survival curves analyzed using the log-rank (Mantel-Cox) test, with time to death / median survival indicated. (FIG. 4E) Lymphocytes; normal reference range indicated by shading. (FIG. 4F) Neutralizing antibody titers. (FIGs. 4B-4C and 4E-4F) Symbols representAttorney Docket No. 10013-110W01individual animals; lines indicate the (FIGs. 4B-4C and 4E) geometric mean or (FIG. 4F) mean except for the vehicle source (n=l) where lines connect individual data points over time. (FIGs. 4B-4C) Period of cohousing indicated by shading, lod, limit of detection.

[0008] FIGs. 5A-5E depict that clinical signs of GHP-88310-prophylaxis in contact animals blocks direct contact transmission of CDV. (FIGs. 5A-5B) Daily (FIG. 5A) temperature and (FIG. 5B) bodyweight measurements. Fever (>39.5°C) indicated by a horizontal dashed line from y-axis. The weight loss endpoint (80%) is shown as a horizontal dashed line from the y-axis. Period of co-housing indicated by shading. (FIGs. 5C-5E) Clinical scores for (FIG. 5C) rash, (FIG. 5D) diarrhea, and (FIG. 5E) conjunctivitis. Symbols represent individual animals; lines represent the mean.

[0009] FIGs. 6A-6E depict that clinical signs of airborne transmission of CDV are prevented or controlled by GHP-88310 treatment of contact animals. (FIGs. 6A-6C) Clinical evaluation of (FIG. 6A) rash, (FIG. 6B) diarrhea, and (FIG. 6C) conjunctivitis after initial challenge. (FIGs. 6D-6E) Daily (FIG. 6D) bodyweight and (FIG. 6E) temperature measurements during both initial and re-challenge. The weight loss threshold (80%) is indicated as a horizontal dashed line from the y-axis. Fever (>39.5°C) is similarly marked with a horizontal dashed line from the axis. (FIGs. 6A-6E) Symbols represent individual animals; lines represent the mean. Initial challenge is denoted by dark shading, re-challenge by light shading.

[0010] FIG. 7 depicts the survival overview of dose down study in ferrets. Viral dose titration study (n=l) assessing three intranasal inoculum concentrations of recombinant CDV, ranging from lxl02to lxlO4TCID50 units. Survival curves analyzed using the log-rank (Mantel-Cox) test, with time to death indicated.

[0011] FIGs. 8A-8H depict an overview of dose down study in ferrets. Viral dose titration study (n=l) assessing three intranasal inoculum concentrations of recombinant CDV, ranging from lxl02to lxlO4TCID50 units. (FIG. 8A) Bodyweight and (FIG. 8B) temperature measured daily. The weight loss threshold (80%) is indicated as a horizontal dashed line from the y-axis. Fever (>39.5°C) is similarly marked with a horizontal dashed line from the axis. Assessment of viral loads in (FIG. 8C) PBMCs and (FIG. 8D) nasal lavage. (FIG. 8E) Lymphocytes. Normal reference range indicated by shading. Evaluation of clinical signs: (FIG. 8F) rash, (FIG. 8G) diarrhea, and (FIG. 8H) conjunctivitis. Symbols represent individual animals, with lines connecting data points for each animal over time, lod, limit of detection.

[0012] FIG. 9A-9E depict that clinical signs of GHP-88310 treatment in CDV-infected ferrets blocks airborne transmission to untreated contacts. (FIGs. 9A-9B) Daily (FIG. 9A) bodyweight and (FIG. 9B) temperature measurements. The weight loss endpoint (80%) is shown as a horizontal dashed line from the y-axis. Fever (>39.5°C) indicated by a horizontal dashed line from y-axis. Period of co-housingAttorney Docket No. 10013-110W01indicated by shading. (FIGs. 9C-9E) Clinical scores for (FIG. 9C) rash, (FIG. 9D) diarrhea, and (FIG. 9E) conjunctivitis. Symbols represent individual animals; lines represent the mean.

[0013] FIGs. 10A-10L depict SAR development of broad-spectrum GHP-class polymerase inhibitors. (FIG.10A) Study schematic of multi-dose GHP-88309 tolerability and PK assessment. Ferrets received 14 oral doses of GHP-88309 at two different concentrations, administered twice daily. (FIG. 10B) Survival curves of animals from (FIG. 10A). (FIG. 10C) PK profile of GHP-88309 (50 mg / kg) in ferret plasma following 14 oral doses. (FIGs. 10D-10F) Survival curves of animals after single oral dose of GHP-88309 (FIG. 10D), body temperature (FIG. 10E), and clinical scoring (FIG. 10F): body temperature (0, >38°C; 1, 37.5-37.9°C; 2, 37-37.4°C; 3, <37°C) and neurologic status (0, explores objects, weasel war dance, intact reflexes; 1, decreased interest in exploring objects; 2, lethargic, and / or altered gait, and / or decreased reflexes; 3, obtunded, and / or seizures, and / or paralysis in one or more limbs). (FIG. 10G) Top-ranking GHP class analogs. (FIG. 10H) Evaluation of cellular viability of GHP-class compounds on Vero-E6 cells. (FIGs. 101- IOK) Recombinant virus reporter-based dose response inhibition of HPIV3 (FIG. 101), MeV (FIG. 10J), and CedV (FIG. 10K), n >3. (FIG. 10L) Study schematic of single-dose GHP-class PK assessment in BALB / c mice. Mice received an oral dose of GHP-class (20 mg / kg) and were monitored over 24 hours. Blood was collected from each sub-group of mice twice, following euthanasia. (FIG. 10M) PK profile of GHP-classes in mouse plasma following a single dose. Group sizes (n) specified in study schematics (FIGs. 10A and IOL). Log-rank (Mantel-Cox) test with median survival shown (FIGs. 10B and 10D). Symbols represent geometric mean ± geometric standard deviation (SD) (FIGs. 10C and 10L). Variable slope 4-parameter non-linear regression modeling (FIGs. 10H-10K), n >3. Symbols represent means ± SD. LLOQ, lower limit of quantitation. AUC, area under the curve.

[0014] FIGs. 11A-11P depict the evaluation of GHP-class compounds in rodents. (FIG. 11A) Study schematic of GHP-class compound efficacy testing in 129xl / SvJ mice. Mice were infected intranasally with 1.5xl05TCIDso units of recombinant SeV. Oral dosing of GHP-class compounds was administered twice daily, starting one day after infection and continuing through 8.5 days post infection. (FIGs. 11B- 11D) Survival curves (FIG. 11B), trachea (FIG. 11C) and lung (FIG. 11D) viral titers determined 5 dpi of animals shown in (FIG. 11A). (FIG. 11E) Study overview. A direct comparison of GHP-88309 and GHP- 88310 efficacy at varying concentrations was conducted in cotton rats. Animals were intranasally infected with 1 xlOsTCID5Ounits of HPIV3 clinical isolate 13J5. Oral dosing was initiated 12 hours post infection and continued b.i.d. (FIGs. 11F-11G) Trachea (FIG. 11F) and combined left and right cranial lungs (FIG. 11G) were collected 3.5 days after infection and viral titers determined by plaque assay. (FIG.11H) Study outline of single-dose PK and tissue distribution analysis in cotton rats, comparing 50 mg / kgAttorney Docket No. 10013-110W01doses of GHP-88309 and GHP-88310. (FIG. 11I-11J) PK profiles of GHP-88309 and GHP-88310 in cotton rat plasma (FIG. Ill) and tissue concentrations of GHP-88309 and GHP-88310 measured 8 hours post dosing (FIG. 11J). Unpaired, two-tailed t-test. Bars indicate the geometric mean ± geometric SD. (FIG. 11K) Overview of a multi-dose GHP-88310 tolerability and PK study. Cotton rats received 14 oral doses of varying concentrations, administered twice daily. (FIGs. 11L-11M) Survival curves (FIG. 11L) and PK profile (FIG. 11M) after 14 doses of GHP-88310. (FIG. 11N) Summary overview of an efficacy evaluation of GHP-88310 administered once daily at different concentrations in cotton rats. Animals were intranasally infected with 1 xlO6TCID50 units of HPIV3, and oral dosing started 12 hours after infection. (FIGs. 11O-11P) Trachea (FIG. 110) and combined left and right cranial lung lobes (FIG. IIP) were collected 3.5 dpi and viral titers determined by plaque assay. In (FIGs. 11B and 11L), log-rank (Mantel- Cox) test with median survival. In (FIGs. 11C, 11D, 11F, 11G, 110, and IIP), bars indicate the geometric mean ± geometric SD, symbols represent individual biological repeats. One-way ANOVA with Tukey's (between group comparisons) or Dunnet's (comparisons to vehicle) post-hoc multiple comparisons test. In (FIGs. Ill and 11M), symbols represent geometric mean ± geometric SD.

[0015] FIGs. 12A-12J depict single and multi-dose tolerability and PK of twice daily GHP-88310 in ferrets. (FIG. 12A) Overview of single oral dose PK study of GHP-88309 and GHP-88310 in ferrets. (FIG.12B) PK profile of GHP-88309 and GHP-88310 in ferret plasma. (FIG. 12C) Schematic of multi-dose tolerability and PK study of GHP-88309 and GHP-88310 at different concentrations. Ferrets received 14 oral doses, administered b.i.d. Maximal plasma concentration (Cmax) was measured daily in ferrets administered 500 and 1,000 mg / kg GHP-88310. (FIG. 12D) Survival curves; data for the 150 mg / kg GHP- 88309 group, originally shown in (FIGs. 10A-10B, are included for direct comparison. Log-rank (Mantel- Cox) test with median survival. (FIG. 12E) Cmaxof GHP-88310 to monitor plasma build-up after doses 1, 2, 4, 6, 8, 10, 12, and 14. (FIG. 12F) PK profile of GHP-88310 in ferret plasma after 14 oral doses. (FIG. 12G) CBC analysis of ferret blood samples quantifying white blood cells (left), red blood cells (middle), and red blood cell width (right). Normal range for each parameter is shown in shading. (Left, middle), symbols indicate the geometric mean ± geometric SD, (right) symbols represent means ± SD. Two-way ANOVA followed by Dunnet's (comparisons to mock) post-hoc multiple comparisons test. (FIGs. 12H-12J) Single dose PK study of GHP-88310 in dogs. Shown are study outline (FIG. 12H), clinical scoring ranks (FIG. 121) showing neurologic status (0, explores objects, intact reflexes; 1, decreased interest in exploring objects; 2, lethargic, and / or altered gait, and / or decreased reflexes; 3, obtunded, and / or seizures, and / or paralysis in one or more limbs), and PK profile of GHP-88310 in dog plasma (FIG. 12J). In (FIGs. 12B, 12E- 12F, and 12J), symbols indicate the geometric mean ± geometric SD.Attorney Docket No. 10013-110W01

[0016] FIGs. 13A-13G depict the therapeutic effect of GHP-88310 on CDV infection in ferrets. (FIG. 13A) Virus yield reduction of recCDV-5804p. Variable slope 4-parameter non-linear regression modeling; n >3. Symbols indicate the geometric mean ± geometric SD. (FIG. 13B) Overview of assessment of q.d. and b.i.d. GHP-88310 therapeutic potential. Ferrets were infected intranasally with 2xl05TCID50 units of recCDV-5804p. Oral administration of GHP-88310 began 3 days after infection and continued until 10 (150 mg / kg) or 21 (50 mg / kg) dpi. (FIG. 13C) Survival curves. Log-rank (Mantel-Cox) test with median survival. (FIGs. 13D-13F) PBMC-associated viremia (FIG. 13 D), nasal lavage viral titers quantified by TCIDsoassay (FIG. 13E), and lymphocyte counts (FIG. 13F) are shown. Normal range of lymphocytes is depicted by shading. (FIG. 13G) Neutralizing antibody titers. In (FIGs. 13D-13F), symbols indicate individual animals and lines represent geometric means. Two-way ANOVA followed by Dunnet's (comparisons to vehicle) post-hoc multiple comparisons test.

[0017] FIGs. 14A-14D depict functional and molecular profiling of GHP-88310. (FIGs. 14A-14C) Doseresponse assays with recombinant virus variants harboring mutations that mediate reduced susceptibility to GHP-88309. Shown are 4-parameter variable slope non-linear regression curves for HPIV3 (FIG. 14A), SeV (FIG. 14B), and MeV (FIG. 14C). Fifty and 90% inhibitory concentrations against each compound are provided. Yellow highlights mark mutations that result in a >5-fold increase in EC50and / or ECgocompared to the unmodified genetic parent virus. (FIG. 14D) In silica docking of GHP-88310. Ribbon representation of the MeV L-P structure (PDBID: 9dus), highlighting the predicted docking location of GHP-88310 (left). Ribbon representation of the top scoring docking pose conserved between GHP-88309 and GHP-88310 (middle). Surface representation of the docking site of GHP-88310 showing the location of the docking pose between the capping and RdRP domains (right). In (FIGs. 14A-14C), curves show 4-parameter variable slope non-linear regression models; n >3. Symbols represent means ± SD (FIGs. 14A and 14C) or the geometric mean ± geometric SD (FIG. 14B).

[0018] FIGs. 15A-15I depict GHP-88310 efficacy in well-differentiated air liquid interface human airway epithelial cultures. (FIG. 15A) Virus yield reduction of HPIV3 (clinical isolate 13J5) in undifferentiated NHBE cells (F3, n=3). (FIG. 15B) Schematic of well-differentiated air-liquid interface human airway epithelial organoids. (FIG. 15C) Assessment of GHP-88310 cytotoxicity in organoids. TEER was measured before and after 48 hours of exposure to increasing concentrations of GHP-88310 in the basolateral chamber. Symbols represent individual biological measurements, and lines show medians. (FIG. 15D) Virus yield reduction assay of HPIV3 in organoids. Shed progeny virus was collected from the apical side of the transwells. Curves show 4-parameter variable slope non-linear regression models, symbols represent the geometric mean ± geometric SD. (FIG. 15E) HPIV3 titers in infected organoids used forAttorney Docket No. 10013-110W01confocal imaging. (FIG. 15F) Confocal microscopy of organoids infected with HPIV3 at 2dpi. Cells were stained with anti-HPIV3 HN, anti-ZO-1 (tight junctions), and Hoechst 34580 (nuclei). Z-stacks of 100-150 0.22 pm slices with 63x oil objective. Scale bar, 10 pm. (FIG. 15G) Ex vivo PK-guided assessment of GHP- 88310 efficacy against HPIV3 in organoids. The simulation recapitulates the PK profile of GHP-88310 following 14 doses of 50 mg / kg b.i.d. (top) or the PK profile after 20 mg / kg q.d. Solid lines represent the experimentally measured PK profiles in ferrets. Corresponding compound concentrations of GHP-88310 applied at each time point are shown as bars. (FIGs. 15H-15I) HPIV3 titers from the ex vivo simulation study. In (FIGs. 15E, 15H, and 151), bars indicate the geometric mean ± geometric SD and symbols represent individual biological repeats. One-way ANOVA with Dunnet's (comparisons to vehicle) post- hoc multiple comparisons test. Group sizes (n) are indicated in each study schematic.

[0019] FIGs. 16A-16C depict multi-dose tolerability of GHP-88309 in ferrets. Bodyweight (FIG. 16A) and body temperature (FIG. 16B). Fever (>39.5°C) is indicated by a horizontal dashed line. (FIG. 16C) CBC analysis of ferret blood samples. Normal range for each parameter is shown in shading. In all panels, symbols represent means ± SD.

[0020] FIGs. 17A-17B depict clinical parameters of SeV infection in mice. (FIG. 17A) Bodyweight.Predefined weight loss endpoint (80%) is shown as a horizontal dashed line. (FIG. 17B) Body temperature. Normal temperature range (36.5 - 38.5°C) is shown as horizontal dashed line. In both panels, symbols represent means ± SD.

[0021] FIGs. 18A-18B depict clinical parameters and viral titers of cotton rats infected with HPIV3.Bodyweight (FIG. 18A) and body temperature (FIG. 18B) of HPIV3-infected animals. Symbols represent means ± SD.

[0022] FIGs. 19A-19B depicts multi-dose tolerability and PK of GHP-88310 in cotton rats. Bodyweight (FIG. 19A) and body temperature (FIG. 19B). Symbols represent means ± SD.

[0023] FIGs. 20A-20B depict once daily administration of GHP-88310 against HPIV3 in cotton rats.Bodyweight (FIGs. 20A) and body temperature (FIGs. 20B) of HPIV3-infected animals. Symbols represent means ± SD.

[0024] FIGs. 21A-21E depict multi-dose tolerability and PK of GHP-88310 in ferrets. (FIGs. 21A-21B) Bodyweight (FIG. 21A) and body temperature (FIG. 21B). Fever (>39.5°C) indicated by a horizontal dashed line. (FIGs. 21C-21E) CBC analysis of ferret blood samples. Normal range for each parameter is indicated in shading. In all panels, symbols represent means ± SD. Two-way ANOVA followed by Dunnet's (comparisons to mock) post-hoc multiple comparisons test.Attorney Docket No. 10013-110W01

[0025] FIG. 22 depicts clinical parameters of multi-dose GHP-88309 and GHP-88310 tolerability. Blood chemistry analysis of serum collected on days 1 or 2, and day 7 after dosing. Serum from the 150 mg / kg GHP-88309 group was collected on day 1 only. Key kidney and liver parameters are highlighted in yellow shading. Values were normalized to those of mock ferrets (n=17) and are presented as percent increase or decrease. Dashed vertical line indicates unchanged to mock-dosed animals (100% mark). Symbols represent individual animals. ALB, albumin; CRE, creatine; BUN, blood urea nitrogen; ALP, alkaline phosphatase; ALT, alanine transaminase; AST, aspartate aminotransferase; TBILC, total bilirubin; GGT, gamma-glutamyl transferase; GLOB, globulins; AMY, amylase; CALA, calcium; CHOL, cholesterol; CK, creatine kinase; PHOS, phosphate; MG, magnesium; TP, total protein; TRIG, triglycerides; NA, sodium; K, potassium; CL, chloride; GLU, glucose; A / G, ratio of albumin to globulin; B / C, ratio of BUN to creatine; NA / K, ratio of sodium to potassium.

[0026] FIGs.23A-23B depict evaluation of once- and twice-daily GHP-88310 dosing for CDV infection in ferrets. Bodyweight (FIG. 23A) and body temperature (FIG. 23B). Predefined weight loss endpoint (80%) and fever (>39.5°C) are shown by horizontal dashed lines. In both panels, symbols represent means ± SD.

[0027] FIGs. 24A-24C depict In silico docking of GHP-88309. (FIG. 24A) Ribbon representation of the MeV L-P structure (PDBID: 9dus) highlighting the docking location of GHP-88309 (black spheres in red dashed box). (FIG. 24B) Ribbon representation of the top scoring docking pose conserved between GHP- 88309 and GHP-88310. (FIG. 24C) Surface representation of the docking site of GHP-88309 showing the location of the predicted docking pose between the capping and RdRP domains.

[0028] FIGs. 25A-25B depict docking poses. 2D-diagram of predicted top-scoring ligand interaction generated with MOE for GHP-88309 (FIG. 25A) and GHP-88310 (FIG. 25B). The predicted arenehydrogen bond interactions between the isoquinoline ring of GHP-88310 and L residue A866 are shown.

[0029] FIG. 26 depicts cellular uptake and retention of GHP-88310 in undifferentiated HBTECs (female donor, F3). Symbols represent means ± SD.

[0030] FIGs. 27A-27B depict confocal imaging of GHP-88310 antiviral activity in human airway epithelium organoids. Confocal microphotographs of organoids infected with HPIV3 were taken 2 dpi. Z- stacks of 100-1500.22 pm slices with 63x oil objective. Scale bar, 10 pm. In (FIG. 27A), cells were stained with anti-PIV3 HN, anti-ZO-1 (tight junctions), and Hoechst 34580 (nuclei). Images shown in (FIG. 15G) are highlighted by a dashed box. In (FIG. 27B), cells were stained with anti-PIV3 HN, anti-MUC5AC (mucin), and Hoechst 34580 (nuclei).Attorney Docket No. 10013-110W01DETAILED DESCRIPTION

[0031] Before the present methods and systems are disclosed and described, it is to be understood that the methods and systems are not limited to specific synthetic methods, specific components, or particular compositions. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0032] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Ranges may be expressed herein as from "about" one particular value, and / or to "about" another particular value. When such a range is expressed, another embodiment includes- from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint and independently of the other endpoint.

[0033] " Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0034] Throughout the description and claims of this specification, the word "comprise" and variations of the word, such as "comprising" and "comprises," mean "including but not limited to," and is not intended to exclude, for example, other additives, components, integers, or steps. " Exemplary" means "an example of" and is not intended to convey an indication of a preferred or ideal embodiment. " Such as" is not used in a restrictive sense, but for explanatory purposes.

[0035] Disclosed are components that can be used to perform the disclosed methods and systems.These and other components are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these components are disclosed, that while specific reference of each various individual and collective combinations and permutations of these may not be explicitly disclosed, each is specifically contemplated and described herein, for all methods and systems. This applies to all aspects of this application including, but not limited to, steps in disclosed methods. Thus, if there are a variety of additional steps that can be performed, it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the disclosed methods.

[0036] Compounds disclosed herein may be provided in the form of acceptable salts, for example, pharmaceutically acceptable salts. Examples of such salts are acid addition salts formed with inorganicAttorney Docket No. 10013-110W01acids, for example, hydrochloric, hydrobromic, sulfuric, phosphoric, and nitric acids and the like; salts formed with organic acids such as acetic, oxalic, tartaric, succinic, maleic, fumaric, gluconic, citric, malic, methanesulfonic, p-toluenesulfonic, napthalenesulfonic, and polygalacturonic acids, and the like; salts formed from elemental anions such as chloride, bromide, and iodide; salts formed from metal hydroxides, for example, sodium hydroxide, potassium hydroxide, calcium hydroxide, lithium hydroxide, and magnesium hydroxide; salts formed from metal carbonates, for example, sodium carbonate, potassium carbonate, calcium carbonate, and magnesium carbonate; salts formed from metal bicarbonates, for example, sodium bicarbonate and potassium bicarbonate; salts formed from metal sulfates, for example, sodium sulfate and potassium sulfate; and salts formed from metal nitrates, for example, sodium nitrate and potassium nitrate.

[0037] The term "alkyl" refers to a radical of a straight-chain or branched hydrocarbon group having a specified range of carbon atoms (e.g., a " Cue alkyl" can have from 1 to 16 carbon atoms). An alkyl group can be a saturated alkyl group or an unsaturated alkyl group, i.e., an alkyl group having one or more carbon-carbon double / triple bonds, i.e., an alkenyl or alkynyl group. Unless specified to the contrary, an "alkyl" group includes both saturated alkyl groups and unsaturated alkyl groups.

[0001] The term "heteroalkyl" refers to an alkyl group, which further includes at least one heteroatom (e.g., 1, 2, 3, or 4 heteroatoms) selected from oxygen, nitrogen, or sulfur within (i.e., inserted between adjacent carbon atoms of) and / or placed at one or more terminal position(s) of the parent chain. By way of example, a heteroCi-ealkyl (which may also be designated a Ci.6heteroalkyl) group includes, but is not limited to, the following structures:

[0002] The term "heteroalkyl" preceded by a separate heteroatom refers to a heteroalkyl group bonded through the specified heteroatom. By way of example, an OCi.6heteroalkyl group includes, but is not limited to, the following structures:

[0003] When a range of values is listed, it is intended to encompass each value and sub-range within the range. For example, " Ci-Salkyl" is intended to encompass Ci, C2, C3, C4, C5, Cs, Ci_6, C1.5, C1.4, C1-3, Ci.2, C2-g, C2-s, C2-4, C2-3, C3.6, C3.5, C3.4, C4.6, C4.5, and C5.6 alkyl.Attorney Docket No. 10013-110W01

[0004] Affixing the suffix "-ene" to a group indicates the group is a polyvalent moiety, e.g., bonded to two or more groups. Alkylene is the polyvalent moiety of alkyl, alkenylene is the divalent moiety of alkenyl, alkynylene is the divalent moiety of alkynyl, heteroalkylene is the divalent moiety of heteroalkyl, heteroalkenylene is the divalent moiety of heteroalkenyl, heteroalkynylene is the divalent moiety of heteroalkynyl, carbocyclylene is the divalent moiety of carbocyclyl, heterocyclylene is the divalent moiety of heterocyclyl, arylene is the divalent moiety of aryl, and heteroarylene is the divalent moiety of heteroaryl (each of which parent groups as defined herein).

[0005] The term "alkoxy" refers to an alkyl group, as defined herein, appended to the parent molecular moiety through an oxygen atom.

[0006] The term "aryl" refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 n electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system (" C6.i4 aryl"). " Aryl" also includes ring systems wherein the aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms continues to designate the number of carbon atoms in the aryl ring system. Unless otherwise specified, each instance of an aryl group is independently unsubstituted (an "unsubstituted aryl") or substituted (a "substituted aryl") with one or more substituents.

[0007] " Aralkyl" is a subset of "alkyl" and refers to an alkyl group substituted by an aryl group, wherein the point of attachment is on the alkyl moiety.

[0008] The term "heteroaryl" refers to a radical of a 5-14 membered monocyclic or polycyclic (e.g., bicyclic, tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 n electrons shared in a cyclic array) having ring carbon atoms and 1-4 ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-14 membered heteroaryl"). In heteroaryl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. Heteroaryl polycyclic ring systems can include one or more heteroatoms in one or both rings. " Heteroaryl" includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the point of attachment is on the heteroaryl ring, and in such instances, the number of ring members continues to designate the number of ring members in the heteroaryl ring system. " Heteroaryl" also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups, wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused polycyclic (aryl / heteroaryl) ring system.Attorney Docket No. 10013-110W01Polycyclic heteroaryl groups wherein one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and the like), the point of attachment can be on either ring, i.e., either the ring bearing a heteroatom (e.g., 2-indolyl) or the ring that does not contain a heteroatom (e.g., 5-indolyl).

[0009] Exemplary heteroaryl and heterocyclyl rings include: benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH carbazolyl, carbolinyl, chromanyl, chromenyL cirrnolinyl, decahydroquinolinyl, 2H,6H~ 1,5,2-dithiazinyl, dihydrofuro[2,3 b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, IH-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5- oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-l,2,5-thiadiazinyl, 1, 2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl, and xanthenyl.

[0010] Unless specified to the contrary, the alkyl, alkenyl, alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups defined herein (and the "ene" versions of said groups) may be substituted or unsubstituted. A substituted group includes a nonhydrogen substituent at a position where, in the unsubstituted version, a hydrogen atom would be found. Substituents include, but are not limited to, halogen, hydroxy, alkyl, alkoxy, nitro, cyano, oxo, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, -NRaRb, -NRaC(=O)Rb, -NRaC(=O)NRaNRb, -NRaC(=O)ORb, - NRaSO2Rb, -C(=O)Ra, -C(=O)ORa, -C(=O)NRaRb, -OC(=O)NRaRb, -ORa, -SRa, -SORa, - S(=O)2Ra, -OS(=O)2Raand -S(=O)2ORa. Raand Rbin this context can be the same or different and independently hydrogen, halogen, hydroxyl, alkyl, alkoxy, alkyl, amino, alkylamino, dialkylamino, carbocyclyl, carbocycloalkyl, heterocarbocyclyl, heterocarbocycloalkyl, aryl, arylalkyl, heteroaryl, or heteroarylalkyl.Attorney Docket No. 10013-110W01

[0011] As used herein, the designation of a polyvalent moiety without specifying the specific order of attachment is intended to cover all possible arrangements. By way of example, a compound represented by the formula:A-X-B,wherein X is NHC(=O) embraces both:0 011 B A 11A^N' 'N^BH and HII

[0012] As used herein, a chemical bond depicted asII represents either a single, double, or triple bond, valency permitting. By way of example,

[0013] Some compounds disclosed herein may exist as one or more tautomers. Tautomers are interconvertible structural isomers that differ in the position of one or more protons or other labile atoms. By way of example:

[0014] Unless stated to the contrary, a substituent drawn without explicitly specifying the point of attachment indicates that the substituent may be attached at any possible atom. For example, in a benzofuran depicted:the substituent may be present at any one of the six possible carbon atoms.

[0015] As used herein, the term "null," when referring to a possible identity of a chemical moiety, indicates that the group is absent, and the two adjacent groups are directly bonded to one another. By way of example, for a genus of compounds having the formula CH3-X-CH3, if X is null, then the resulting compound has the formula CH3-CH3. A group having the subscript '0' is understood to represent a nullAttorney Docket No. 10013-110W01group as well. By way of example, in the compound CH3-(X)Z-CH3, if X is CH2and z is 0, then the compound has the formula CH3-CH3.

[0016] A bracketed functional group with a subscripted variable should be understood to denote the number of repeated bracketed groups present. For example, a number that is selected from 0 or 1 should be interpreted as follows:a = 0 = Ri-O-R2

[0017] In certain instances, two or more variable groups may together form a ring. It is understood that any depicted atoms separated from the identified groups will themselves form part of the ring:*3 *3When the variable groups are substituted on an aromatic system, the new ring will be a fused ring, and unless specified to the contrary, may be either aromatic or non-aromatic, carbocyclic or heterocyclic:The ring may further be defined by the number of carbon atoms in the specific ring formed by the variable groups, which includes the atoms separating the variable groups:aC6aryl C5heterocyclyl C5heteroaryl Each of the above results occurs when R1and R2together form a six-membered (or six-atom) ring. Other rings, including 3, 4, 5, 7, and 8-member rings, may also be formed and may be further limited by a specified number of carbon atoms. Although the singular "a ring" may be used to define the group, unless specified to the contrary, both monocyclic and polycyclic rings are possible:2 monring polycyclic ring

[0018] Unless stated to the contrary, a formula with chemical bonds shown only as solid lines and not as wedges or dashed lines contemplates each possible isomer, e.g., each enantiomer, diastereomer, andAttorney Docket No. 10013-110W01meso compound, and a mixture of isomers, such as a racemic or scalemic mixture. Unless stated to the contrary, a formula depicting one or more stereochemical features does not exclude the presence of other isomers.

[0019] As used herein, the chemical group " D" refers to deuterium at an isotopic abundance greater than 25%, 35%, 50%, 60%, 70%, 80%, or 90%. In certain implementations, the chemical group " D" refers to deuterium at an isotopic abundance greater than 50%.

[0020] As used herein, the compound references " EIDD-3608 / GHP-88310", " EIDD-3608," and " GHP- 88310” can be used interchangeably and refer to a compound having a structure given by the following formula:

[0021] Disclosed herein are compounds of Formula (1):[Formula (1)],or a pharmaceutically acceptable salt thereof, whereinX1is N or CRX1; wherein RX1is selected from H, D, halo, COOH, CO2Ci.6alkyl, CN, Ci.6a Ikyl, Ci- sheteroalkyl, C3-6cycloalkyl, or Ci.gheterocyclyl;X2is N or CRx2; wherein Rx2is selected from H, D, halo, COOH, CO2C1-6alkyl, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;X3is N or CRx3; wherein Rx3is selected from H, D, halo, COOH, CO2C1-6alkyl, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;wherein at least one of X1, X2, or X3is N;Y1is N or CRyl; wherein Ry1is selected from H, D, halo, COOH, CO2C1-6alkyl, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;Attorney Docket No. 10013-110W01Y2is N or CRy2; wherein Ry2is selected from H, D, halo, COOH, CO2C1-6alkyl, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;Y3is N or CRy3; wherein Ry3is selected from H, D, halo, COOH, CO2C1-6alkyl, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;Y4is N or CRy4; wherein Ry4is selected from H, D, halo, COOH, CO2C1-6alkyl, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;wherein at least one of Y1, Y2, Y3, or Y4is N;Z is C(=O) or SO2,R1is H or C1-6alkyl;R2is H or C1-6alkyl;Rais selected from H, D, halo, COOH, CO2C1-6alkyl, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;Rbis selected from H, D, halo, CO2C1-6alkyl, COOH, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;Rcis selected from H, D, halo, CO2C1-6alkyl, COOH, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;Rdis selected from H, D, halo, COOH, CO2C1-6alkyl, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;wherein any two or more of Rx1, Rx2, Rx3, Ry1, Ry2, Ry3, Ry4, R1, R2, Ra, Rb, Rc, and Rdmay together form a ring.

[0022] In some implementations, the compound can be a compound of Formula (2a), (2b), (2c), or (2d):Attorney Docket No. 10013-110W01

[0023] In some implementations of the compounds of Formula (1), (2a), (2b), (2c), and (2d), one of Y1, Y2, Y3, orY4is N.

[0024] In some implementations, the compound can be a compound of Formula (3a), (3b), (3c), or (3d):[Formula (3b)], Ryi[Formula (3d)].

[0025] In some implementations of the compounds of (3a), (3b), (3c), and (3d), one of X1, X2, and X3is N, and the other two are CH.

[0026] In some implementations, the compound can be a compound of Formula (4a), (4b), (4c), or (4d):Attorney Docket No. 10013-110W01Rx3Ryi[Formula (4a)], [Formula (4b)],

[0027] In some implementations, Rv4and Rdtogether form a ring, for example, a compound of Formula (5):[Formula (5)],wherein Q1is null, O, S, NRn, and Rnis H or Ci-3alkyl, C1-3alkylene, or C1-3heteroalkylene.

[0028] In certain implementations, the compound can be a compound of Formula (6a), (6b), (6c), (6d), or (6e):Attorney Docket No. 10013-110W01[Formula (6a)], [Formula (6b)],[Formula (6c)], [Formula (6d)], or

[0029] In some implementations, the compound can be a compound of Formula (7a), (7b), (7c), or (7d):[Formula (7a)]. [Formula (7b)].Attorney Docket No. 10013-110W01[Formula (7c)], or [Formula (7d)].

[0030] In some implementations, R2and Ratogether form a ring, for example, a compound of Formula (8a), (8b), (8c):[Formula (8b)], or[Formula (8c)].

[0031] In some implementations, the compound can be a compound of Formula (9a), (9b), or (9c):Attorney Docket No. 10013-110W01Ryi Ryi[Formula (9a)], [Formula (9b)], orRx3Ry1[Formula (9c)].

[0032] In some implementations, R1and R2can both be H, and the compound can be a compound of Formula (10a), (10b), or (10c):Ryi Ryi[Formula (10a)], [Formula (10b)], orAttorney Docket No. 10013-110W01Rx3Ryi[Formula (10c)].

[0033] In some implementations, R1and R2can both be H, and two or more of Rx1, Rx3, Ryl, Ry3, and Ry4can be H, and the compound can be a compound of Formula (11a), (lib), or (11c):[Formula (11c)].

[0034] In some implementations, the compound can be a compound of formula (12a) or (12b):Attorney Docket No. 10013-110W01[Formula (12b)];wherein Q is selected from S or O; andRa10and Ra11are independently selected from C1-6alkyl.

[0035] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (7a), (7b), (7c), (7d), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), (11c), (12a) and (12b) Rdis H.

[0036] In some implementations, Raand Rbtogether form a ring, or Rband Rctogether form a ring. In some implementations, Rdis H, and either Raand Rbtogether form a ring or Rband Rctogether form a ring. The ring may be a carbocyclic ring or a heterocyclic ring. In some implementations, the ring is a heterocyclic ring having 1, 2, or 3 annular heteroatoms. In some implementations, the ring is a heterocyclic ring having 2 annular heteroatoms, for example, an acetal. In some implementations, the compound is a compound of Formula (13a) or (13b):wherein R3and R3are independently selected from H, D, halo, C1-3alkyl, C1-3heteroalkyl, or R3and R3together form an oxo, a C3-6cycloalkyl, or a C1-6heterocyclyl; andR4and R4are independently selected from H, D, halo, C1-3alkyl, C1-3heteroalkyl, or R4and R4together form an oxo, a C3.6cycloalkyl, or a Ci-eheterocyclyl.Attorney Docket No. 10013-110W01

[0037] In some implementations, R1and R2are each H.

[0038] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), R1, R2, Rb, Rc, and Rdare each H and Rais H, D, C3. ecycloalkyl, Ci-6alkyl, halo, or Ci-6heteroalkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais H, F, cyclopropyl, cyclobutyl, methyl, ethyl, methoxy, ethoxy, thiomethyl, thioethyl, methylamino, dimethylamino, ethylamino, or diethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais not H.

[0039] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais D. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais C3. ecycloalkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais Ci.6alkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais halo. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais Ci.6heteroalkyl.

[0040] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais F. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais cyclopropyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais cyclobutyl. In some implementations of theAttorney Docket No. 10013-110W01compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais methyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (11b), and (11c), Rais ethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais methoxy. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais ethoxy. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais thiomethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (Ila), (lib), and (11c), Rais thioethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais methylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais dimethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (11b), and (11c), Rais ethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais diethylamino.

[0041] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), R1, R2, Ra, Rc, and Rdare each H and Rbis H, D, C3-6cycloalkyl, C1-6alkyl, halo, or C1-6heteroalkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais H, F,Attorney Docket No. 10013-110W01cyclopropyl, cyclobutyl, methyl, ethyl, methoxy, ethoxy, thiomethyl, thioethyl, methylamino, dimethylamino, ethylamino, or diethylamino In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis not H.

[0042] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis D. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis C3-6cycloalkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (11b), and (11c), Rbis C1-6alkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis halo. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis Ci-Sheteroalkyl.

[0043] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis F. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis cyclopropyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis cyclobutyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis methyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis ethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), andAttorney Docket No. 10013-110W01(11c), Rbis methoxy. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis ethoxy. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis thiomethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (Ila), (lib), and (11c), Rbis thioethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis methylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis dimethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis ethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rbis diethylamino.

[0044] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), R1, R2, Ra, Rb, and Rdare each H and Rcis H, D, C3-6cycloalkyl, C1-6alkyl, halo, or C1-6heteroalkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (11b), and (11c), Rais H, F, cyclopropyl, cyclobutyl, methyl, ethyl, methoxy, ethoxy, thiomethyl, thioethyl, methylamino, dimethylamino, ethylamino, or diethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis not H.

[0045] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis D. In some implementations of the compound ofAttorney Docket No. 10013-110W01Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis C3-6cycloalkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (11b), and (11c), Rcis C1-6alkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis halo. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis C1-6heteroalkyl.

[0046] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis F. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis cyclopropyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis cyclobutyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis methyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (11b), and (11c), Rcis ethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis methoxy. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis ethoxy. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis thiomethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a),Attorney Docket No. 10013-110W01(8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis thioethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis methylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis dimethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis ethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rcis diethylamino.

[0047] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), R1, R2, Ra, Rb, and Rcare each H and Rdis H, D, C3. ecycloalkyl, Ci-ealkyl, halo, or C1-6heteroalkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rais H, F, cyclopropyl, cyclobutyl, methyl, ethyl, methoxy, ethoxy, thiomethyl, thioethyl, methylamino, dimethylamino, ethylamino, or diethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis not H.

[0048] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis D. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis C3. ecycloalkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis Ci.6alkyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), andAttorney Docket No. 10013-110W01(11c), Rdis halo. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis C1-6heteroalkyl.

[0049] In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis F. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (Ila), (lib), and (11c), Rdis cyclopropyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis cyclobutyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis methyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis ethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis methoxy. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis ethoxy. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis thiomethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis thioethyl. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis methylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (11b), and (11c), Rdis dimethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a),Attorney Docket No. 10013-110W01(4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis ethylamino. In some implementations of the compound of Formula (1), (2a), (2b), (2c), (2d), (3a), (3b), (3c), (3d), (4a), (4b), (4c), (4d), (5), (6a), (6b), (6c), (6d), (6e), (7a), (7b), (7c), (7d), (8a), (8b), (8c), (9a), (9b), (9c), (10a), (10b), (10c), (11a), (lib), and (11c), Rdis diethylamino.

[0050] In some implementations, the compound has the formula:Attorney Docket No. 10013-110W01Attorney Docket No. 10013-110W01R1[Formula (14o)] [Formula (14p)],[Formula (14q)],d^alkyl [Formula (14s)],R1H2Ni^alkyl 6-|.3alkyl [Formula (14u)], [Formula (14va)],Attorney Docket No. 10013-110 WO 161-3alkyl [Formula (14w)], 1-3a^yl [Formula ( 14x)],IxC1.3alkyl PC1.3alkylC-|_3alkyl [Formula (14y)], 1-3alky! [Formula (14z)],[Formula (14bb)],[Formula (14dd)],Attorney Docket No. 10013-110W01[Formula (14kk)], [Formula (1411)],[Formula (14mm)], [Formula (14nn)],Attorney Docket No. 10013-110W01[Formula (14oo)], or [Formula (14pp)].

[0051] In some implementations of the compounds of formula (14a)-(14pp), Y2is N, and Y1, Y3, and Y4are CH. In some implementations of the compounds of formula (14a)-(14pp), Y2is N, Y1, Y3, and Y4are CH, X1is N, and X2and X3are CH.

[0052] In some implementations of the compounds of formula (14a)-(14pp), Y2is N, Y1, Y3, and Y4are CH, X2is N, and X1and X3are CH.

[0053] In some implementations of the compounds of formula (14a)-(14pp), Y2is N, Y1, Y3, and Y4are CH, X3is N, and X1and X2are CH.

[0054] In certain implementations, the compound is a compound selected from,or a pharmaceutically acceptable salt thereof.Attorney Docket No. 10013-110W01

[0055] Also disclosed herein are methods of preparing the disclosed compounds. In some implementations, a metal-catalyzed cross-coupling can be performed with compounds having the formula:wherein Ra- Rb, Rc, R^X1, X2, X3, Y1, Y2, Y3, and Y4are as defined herein, and X and Y are complementary cross-coupling functional groups. In some implementations, one of X and Y is a boronic acid, boronic ester, trialkyltin (e.g., tributyltin or trimethyl tin), Zn-halide, organosilane (e.g., Si(Me)3, Si(Me)2OK, SSiF3), or COOH, and the other is halide (e.g., F, Cl, Br, or I) or sulfonate (e.g., triflate or mesylate). The nitrile in the cross-coupled product may then be hydrolyzed to provide an amide. Persons skilled in the art will recognize that the disclosed compounds may be prepared using other chemistries as well.Moreover, any of RaRb, Rcand Rd, Rx1, Rx2, Rx3, Rvl, Rv2, Ry3, and RX1may be provided in a suitable protected form for subsequent conversion to other groups.

[0056] Certain materials, compounds, compositions, and components used in the above methods can be obtained commercially or readily synthesized using techniques generally known to those of skill in the art. For example, the starting materials and reagents used in preparing the disclosed compounds and compositions are either available from commercial suppliers, such as Sigma-Aldrich (formerly MilliporeSigma, Burlington, MA) or Thermo Fisher Scientific Inc. (Waltham, MA), or are prepared by methods known to those skilled in the art following procedures set forth in references such as Fieser and Fieser's Reagents for Organic Synthesis (John Wiley and Sons, 2007); Organic Reactions (John Wiley and Sons, 2004); March's Advanced Organic Chemistry, (John Wiley and Sons, 8thEdition); and Larock's Comprehensive Organic Transformations (John Wiley and Sons, 3rdedition, 2017).

[0057] Variations on compounds used in the above methods can include the addition, subtraction, or movement of various constituents as described for each compound. Similarly, when one or more chiral centers are present in a molecule, the chirality of the molecule can be changed. Additionally, the synthesis of the compounds used in these processes can involve the protection of various chemical groups, and further, the compounds prepared by the disclosed processes may be subsequently deprotected as appropriate. The use of protection and deprotection, and the selection of appropriate protecting groups, would be readily known to one skilled in the art. " Protecting group," as used herein, refers to any conventional functional group that allows one to obtain chemoselectivity in a subsequentAttorney Docket No. 10013-110W01chemical reaction. Protecting groups are described, for example, in Peter G. M. Wuts, Greene's Protective Groups in Organic Synthesis, 5thEd., Wiley & Sons, 2014. For a particular compound and / or a particular chemical reaction, a person skilled in the art knows how to select and implement appropriate protecting groups and their associated synthetic methods. Examples of amine protecting groups include acyl and alkoxy carbonyl groups, such a t-butoxycarbonyl (BOC) and [2-(trimethylsilyl)ethoxy]methoxy (SEM). Examples of carboxyl protecting groups include Ci-C6alkoxy groups, such as methyl, ethyl, and t- butyl. Examples of alcohol protecting groups include benzyl, trityl, silyl ethers, and the like.

[0058] The above methods can be carried out in solvents indicated herein, or in solvents which can be selected by one of skill in the art of organic synthesis. Solvents can be substantially nonreactive with the starting materials (reactants), intermediates, or products under the conditions at which the reaction is carried out, i.e., temperature and pressure. Reactions can be carried out in one solvent or a mixture of more than one solvent. Product or intermediate formation can be monitored according to any suitable method known in the art. For example, product formation can be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g.,1H and13C), infrared spectroscopy, spectrophotometry (e.g., UV-visible), or mass spectrometry, or by chromatography such as high- performance liquid chromatography (HPLC) or thin-layer chromatography (TLC).

[0059] Also disclosed herein are pharmaceutical compositions including one or more of the disclosed compounds. The pharmaceutical compositions typically include an effective amount of a disclosed compound and a pharmaceutically acceptable excipient or an effective amount of a disclosed compound and a suitable pharmaceutical acceptable carrier. The preparations may be prepared in a manner known per se, which usually involves mixing the at least one compound according to the disclosure with the one or more pharmaceutically acceptable carriers, and, if desired, in combination with other pharmaceutical active compounds, when necessary, under aseptic conditions. Reference is made to U. S. Pat. No. 6,372,778, U. S. Pat. No. 6,369,086, U. S. Pat. No. 6,369,087 and U. S. Pat. No. 6,372,733 and the further references mentioned above, as well as to the standard handbooks, such as the latest edition of Remington's Pharmaceutical Sciences.

[0060] Generally, for pharmaceutical use, the compounds may be formulated as a pharmaceutical preparation comprising at least one compound and at least one pharmaceutically acceptable carrier, diluent, or excipient, and optionally one or more further pharmaceutically active compounds.

[0061] The pharmaceutical preparations of the disclosure are preferably in a unit dosage form, and may be suitably packaged, for example in a box, blister, vial, bottle, sachet, ampoule or in any other suitable single-dose or multi-dose holder or container (which may be properly labeled); optionally with one orAttorney Docket No. 10013-110W01more leaflets containing product information and / or instructions for use. Generally, such unit dosages will contain between 1 and 1000 mg, and usually between 5 and 500 mg, of the at least one compound of the disclosure, e.g., about 10, 25, 50, 100, 200, 300 or 400 mg per unit dosage.

[0062] The compounds can be administered by a variety of routes, including the oral, ocular, rectal, transdermal, subcutaneous, sublingual, intravenous, intramuscular, or intranasal routes, depending mainly on the specific preparation used. The compound will generally be administered in an "effective amount," by which is meant any amount of a compound that, upon suitable administration, is sufficient to achieve the desired therapeutic or prophylactic effect in the subject to which it is administered. Usually, depending on the condition to be prevented or treated and the route of administration, such an effective amount will usually be between 0.01 to 1000 mg per kilogram body weight of the patient per day, every other day, twice weekly, or weekly, more often between 0.1 and 500 mg, such as between 1 and 250 mg, for example about 5, 10, 20, 50, 100, 150, 200 or 250 mg, per kilogram body weight of the patient per day, every other day, twice weekly, or weekly, which may be administered as a single daily, every other day, twice weekly, or weekly dose, or divided over one or more daily, every other day, twice weekly, or weekly doses. The amount(s) to be administered, the route of administration, and the further treatment regimen may be determined by the treating clinician, depending on factors such as the age, gender, and general condition of the patient and the nature and severity of the disease / symptoms to be treated. Reference is made to U. S. Pat. No. 6,372,778, U. S. Pat. No. 6,369,086, U. S. Pat. No. 6,369,087 and U. S. Pat. No. 6,372,733 and the further references mentioned above, as well as to the standard handbooks, such as the latest edition of Remington's Pharmaceutical Sciences.

[0063] For an oral administration form, the compound can be mixed with suitable additives, such as excipients, stabilizers, or inert diluents, and brought by means of the customary methods into the suitable administration forms, such as tablets, coated tablets, hard capsules, aqueous, alcoholic, or oily solutions. Examples of suitable inert carriers are gum arabic, magnesia, magnesium carbonate, potassium phosphate, lactose, glucose, or starch, in particular, cornstarch. In this case, the preparation can be carried out both as dry and as moist granules. Suitable oily excipients or solvents are vegetable or animal oils, such as sunflower oil or cod liver oil. Suitable solvents for aqueous or alcoholic solutions are water, ethanol, sugar solutions, or mixtures thereof. Polyethylene glycols and polypropylene glycols are also useful as further auxiliaries for other administration forms. As immediate release tablets, these compositions may contain microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate, lactose, and / or other excipients, binders, extenders, disintegrants, diluents, and lubricants known in the art.Attorney Docket No. 10013-110W01

[0064] When administered by nasal aerosol or inhalation, the compositions may be prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and / or other solubilizing or dispersing agents known in the art. Suitable pharmaceutical formulations for administration in the form of aerosols or sprays are, for example, solutions, suspensions, or emulsions of the compounds of the disclosure or their physiologically tolerable salts in a pharmaceutically acceptable solvent, such as ethanol or water, or a mixture of such solvents. If required, the formulation may additionally contain other pharmaceutical auxiliaries such as surfactants, emulsifiers, and stabilizers as well as a propellant.

[0065] For subcutaneous or intravenous administration, the compounds, if desired, with the substances customary for such purposes, such as solubilizers, emulsifiers, or further auxiliaries, are brought into solution, suspension, or emulsion. The compounds may also be lyophilized and the lyophilizates obtained used, for example, for the production of injection or infusion preparations. Suitable solvents are, for example, water, physiological saline solution, or alcohols, e.g., ethanol, propanol, glycerol, sugar solutions such as glucose or mannitol solutions, or mixtures of the various solvents mentioned. The injectable solutions or suspensions may be formulated according to known art, using suitable non-toxic, parenterally-acceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.

[0066] When rectally administered in the form of suppositories, the formulations may be prepared by mixing the compounds of formula I with a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters, or polyethylene glycols, which are solid at ordinary temperatures, but liquefy and / or dissolve in the rectal cavity to release the drug.

[0067] In certain embodiments, it is contemplated that these compositions can be extended-release formulations. Typical extended-release formulations utilize an enteric coating. Typically, a barrier is applied to oral medication that controls the location in the digestive system where it is absorbed.Enteric coatings prevent the release of medication before it reaches the small intestine. Enteric coatings may contain polymers of polysaccharides, such as maltodextrin, xanthan, scleroglucan dextran, starch, alginates, pullulan, hyaloronic acid, chitin, chitosan and the like; other natural polymers, such as proteins (albumin, gelatin etc.), poly-L-lysine; sodium poly(acrylic acid); poly(hydroxyalkylmethacrylates) (for example poly(hydroxyethylmethacrylate)); carboxypolymethylene (for example Carbopol™); carbomer; polyvinylpyrrolidone; gums, such as guar gum, gum arabic, gum karaya, gum ghatti, locustAttorney Docket No. 10013-110W01bean gum, tamarind gum, gellan gum, gum tragacanth, agar, pectin, gluten and the like; poly(vinyl alcohol); ethylene vinyl alcohol; polyethylene glycol (PEG); and cellulose ethers, such as hydroxymethylcellulose (HMC), hydroxyethylcellulose (HEC), hydroxypropylcellulose (HPC), methylcellulose (MC), ethylcellulose (EC), carboxyethylcellulose (CEC), ethylhydroxyethylcellulose (EHEC), carboxymethylhydroxyethylcellulose (CMHEC), hydroxypropylmethyl-cellulose (HPMC), hydroxypropylethylcellulose (HPEC) and sodium carboxymethylcellulose (Na-CMC); as well as copolymers and / or (simple) mixtures of any of the above polymers. Certain of the above-mentioned polymers may further be crosslinked by way of standard techniques.

[0068] The choice of polymer will be determined by the nature of the active ingredient / drug that is employed in the composition of the disclosure, as well as the desired rate of release. In particular, it will be appreciated by the skilled person, for example, in the case of HPMC, that a higher molecular weight will, in general, provide a slower rate of release of drug from the composition. Furthermore, in the case of HPMC, different degrees of substitution of methoxyl groups and hydroxypropoxyl groups will give rise to changes in the rate of release of the drug from the composition. In this respect, and as stated above, it may be desirable to provide compositions of the disclosure in the form of coatings in which the polymer carrier is provided by way of a blend of two or more polymers of, for example, different molecular weights in order to produce a particular required or desired release profile.

[0069] Microspheres of polylactide, polyglycolide, and their copolymer poly(lactide-co-glycolide) may be used to form sustained-release protein delivery systems. Proteins can be entrapped in the poly(lactide-co-glycolide) microsphere depot by a number of methods, including formation of a water- in-oil emulsion with water-borne protein and organic solvent-borne polymer (emulsion method), formation of a solid-in-oil suspension with solid protein dispersed in a solvent-based polymer solution (suspension method), or by dissolving the protein in a solvent-based polymer solution (dissolution method). One can attach poly(ethylene glycol) to proteins (PEGylation) to increase the in vivo half-life of circulating therapeutic proteins and decrease the chance of an immune response.

[0070] Liposomal suspensions (including liposomes targeted to viral antigens) may also be prepared by conventional methods to produce pharmaceutically acceptable carriers. This may be appropriate for the delivery of free nucleosides, acyl nucleosides, or phosphate ester prodrug forms of the nucleoside compounds according to the present invention.

[0071] The disclosed compounds may be used to treat or prevent a viral infection. The disclosed compounds may be given to a subject in need thereof that has been, or potentially will be, exposed to a virus. The disclosed compounds may be given prior to the development of symptoms. In otherAttorney Docket No. 10013-110W01implementations, the disclosed compounds may be given subsequent to the development of symptoms, for instance, within 1 day of exhibiting symptoms, within 2 days of exhibiting symptoms, within 3 days of exhibiting symptoms, or within 4 days of exhibiting symptoms. In some implementations, the disclosed compounds can be given after the subject has exhibited symptoms. For example, the disclosed compounds may be first given 1 day after exhibiting symptoms, 2 days after exhibiting symptoms, 3 days after exhibiting symptoms, 4 days after exhibiting symptoms, 5 days after exhibiting symptoms, 6 days after exhibiting symptoms, 7 days after exhibiting symptoms, 8 days after exhibiting symptoms, 9 days after exhibiting symptoms, or 10 days after exhibiting symptoms.

[0072] In some implementations, the subject is a human. In some implementations, the subject is a human of 18 years of age or less. In some implementations, the subject is a human who is at least 60 years old or at least 65 years old. In some implementations, the subject is a livestock animal, for example, a cow, pig, goat, sheep, chicken, or turkey. In some implementations, the subject is a domesticated animal, for example, a cat or dog. In some implementations, the subject is a bird.

[0073] The disclosed compounds may be provided to the subject by a number of different administration routes. In some implementations, the disclosed compounds can be administered orally, parenterally, inhalationally, intranasally, or topically, including by eye drops.

[0074] In certain implementations, the disclosed compounds may be used to prevent viral transmission from infected animals to untreated animals. In certain implementations, the disclosed compounds may be used to prevent viral transmission from infected animals to untreated sentinels.

[0075] In some implementations, the disclosed compounds may be used to treat or prevent a paramyxovirus infection (as defined by ICTV Virus Taxonomy Profile: Paramyxoviridae in https: / / doi. Org / 10.1099 / jgv.0.001328). In some implementations, the disclosed compounds may be used to treat or prevent an orthoparamyxovirus infection. In some implementations, the disclosed compounds may be used to treat or prevent a respirovirus infection, a morbillivirus infection, or a henipavirus infection. In some implementations, the disclosed compounds may be used to treat or prevent a parainfluenza virus. In some implementations, the disclosed compounds may be used to treat or prevent a human parainfluenza virus. In some implementations, the disclosed compounds may be used to treat or prevent a terrestrial morbilivirus. In some implementations, the disclosed compounds may be used to treat or prevent a human parainfluenza virus type 1 or human parainfluenza virus type 3. In some implementations, the disclosed compounds may be used to treat or prevent measles virus. In some implementations, the disclosed compounds may be used to treat or prevent canine distemper virus.Attorney Docket No. 10013-110W01

[0076] In some implementations, the disclosed compounds may be used to treat a viral infection. In some implementations, the disclosed compounds may be used to treat a paramyxovirus infection. In some implementations, the disclosed compounds may be used to treat an orthoparamyxovirus infection. In some implementations, the disclosed compounds may be used to treat a respirovirus infection, a morbillivirus infection, or a henipavirus infection. In some implementations, the disclosed compounds may be used to treat a parainfluenza virus. In some implementations, the disclosed compounds may be used to treat a human parainfluenza virus. In some implementations, the disclosed compounds may be used to treat a terrestrial morbilivirus. In some implementations, the disclosed compounds may be used to treat a human parainfluenza virus type 1 or human parainfluenza virus type 3. In some implementations, the disclosed compounds may be used to treat measles virus. In some implementations, the disclosed compounds may be used to treat canine distemper virus.

[0077] Disclosed are methods of treating or preventing a viral infection comprising administering to a subject in need thereof an effective amount of a disclosed compound, or a pharmaceutical composition comprising a compound of a disclosed compound and a pharmaceutically acceptable excipient; wherein the subject is administered a loading dose of the pharmaceutical composition in a first treatment period; and wherein the subject is administered a treatment dose of the pharmaceutical composition in a second treatment period following the first treatment period.

[0078] Also disclosed are methods of treating or preventing a viral infection comprising administering to a subject in need thereof an effective amount of a disclosed compound, or a pharmaceutical composition comprising a compound of A disclosed compound and a pharmaceutically acceptable excipient; wherein the subject is administered a loading dose of the pharmaceutical composition in a first treatment period; and wherein the subject is administered a treatment dose of the pharmaceutical composition in a second treatment period following the first treatment period.

[0079] In exemplary embodiments, the first treatment period is days 1-5 following diagnosis of the viral infection or presentation for preventing the viral infection; the first treatment period is days 1-2 following diagnosis of the viral infection or presentation for preventing the viral infection; the first treatment period is day 1 following diagnosis of the viral infection or presentation for preventing the viral infection; and other periods as encompassed by the foregoing.

[0080] In exemplary embodiments, the first treatment period is 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 18 hours, or 24 hours prior to infection or exposure to a virus, and the second treatment period is a period of treatment following infection comprising 1-7 days of treatment following infection, e.g., for 1Attorney Docket No. 10013-110W01day, 2 days, 3 days, 4 days, 5 days, 6 days, or 7 days after initial viral infection. As disclosed elsewhere herein, the dosing following infection may be daily, every other day, and the like.

[0081] In exemplary embodiments, the loading dose is about 1.1-fold to about 10-fold the treatment dose; the loading dose is about 1.5-fold to about 5-fold the treatment dose; the loading dose is about 1.5-fold to about 2.5-fold the treatment dose; and other loading doses as encompassed by the foregoing.

[0082] In exemplary embodiments, the loading dose is administered once daily, two times daily, three times daily, or four times daily, or alternatively, every other day, every third day, and the like, and other periods of administration as contemplated within the foregoing. In some embodiments, the loading dose is administered at least twice daily. In further embodiments, the loading dose is divided equally among the number of times administered daily. The daily dosing of a loading dose, as described in the foregoing regarding doses per day, can be repeated and occur for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more days.

[0083] In exemplary embodiments, the treatment is delayed or late following initial diagnosis or presentation of the viral infection, that is, treatment initiates after some number of days after diagnosis or presentation of the viral infection, e.g., at days 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more, and then continues for a suitable period of time, e.g., for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more days after initiation.

[0084] In some implementations, the disclosed compounds may be administered in combination with one or more additional antiviral agents. The combination therapy may provide "synergy" and "synergistic effect", i.e., the effect achieved when the active ingredients used together is greater than the sum of the effects that result from using the compounds separately. A synergistic effect may be attained when the active ingredients are: (1) co-formulated and administered or delivered simultaneously in a combined formulation; (2) delivered by alternation or in parallel as separate formulations; or (3) by some other regimen. When delivered in alternation therapy, a synergistic effect may be attained when the compounds are administered or delivered sequentially, e.g., in separate tablets, pills, or capsules, or by different injections in separate syringes. In general, during alternation therapy, an effective dosage of each active ingredient is administered sequentially, i.e., serially, whereas in combination therapy, effective dosages of two or more active ingredients are administered together.

[0085] In some implementations, the disclosed compounds can be administered adjunctively with other active compounds. These compounds include but are not limited to analgesics, anti-inflammatory drugs, antipyretics, antidepressants, antiepileptics, antihistamines, antimigraine drugs, antimuscarinics, anxiolytics, sedatives, hypnotics, antipsychotics, bronchodilators, anti-asthma drugs, cardiovascularAttorney Docket No. 10013-110W01drugs, corticosteroids, dopaminergics, electrolytes, gastro-intestinal drugs, muscle relaxants, nutritional agents, vitamins, parasympathomimetics, stimulants, anorectics, anti-narcoleptics, and antiviral agents. " Adjunctive administration," as used herein, means the compound can be administered in the same dosage form or in separate dosage forms with one or more other active agents. The additional active agent(s) can be formulated for immediate release, controlled release, or combinations thereof.

[0086] Specific examples of compounds that can be adjunctively administered with the compounds include, but are not limited to, aceclofenac, acetaminophen, adomexetine, almotriptan, alprazolam, amantadine, amcinonide, aminocyclopropane, amitriptyline, amolodipine, amoxapine, amphetamine, aripiprazole, aspirin, atomoxetine, azasetron, azatadine, beclomethasone, benactyzine, benoxaprofen, bermoprofen, betamethasone, bicifadine, bromocriptine, budesonide, buprenorphine, bupropion, buspirone, butorphanol, butriptyline, caffeine, carbamazepine, carbidopa, carisoprodol, celecoxib, chlordiazepoxide, chlorpromazine, choline salicylate, citalopram, clomipramine, clonazepam, clonidine, clonitazene, clorazepate, clotiazepam, cloxazolam, clozapine, codeine, corticosterone, cortisone, cyclobenzaprine, cyproheptadine, demexiptiline, desipramine, desomorphine, dexamethasone, dexanabinol, dextroamphetamine sulfate, dextromoramide, dextropropoxyphene, dezocine, diazepam, dibenzepin, diclofenac sodium, diflu nisal, dihydrocodeine, dihydroergotamine, dihydromorphine, dimetacrine, divalproxex, dizatriptan, dolasetron, donepezil, dothiepin, doxepin, duloxetine, ergotamine, escitalopram, estazolam, ethosuximide, etodolac, femoxetine, fenamates, fenoprofen, fentanyl, fludiazepam, fluoxetine, fluphenazine, flurazepam, flurbiprofen, flutazolam, fluvoxamine, frovatriptan, gabapentin, galantamine, gepirone, ginko bilboa, granisetron, haloperidol, huperzine A, hydrocodone, hydrocortisone, hydromorphone, hydroxyzine, ibuprofen, imipramine, indiplon, indomethacin, indoprofen, iprindole, ipsapirone, ketaserin, ketoprofen, ketorolac, lesopitron, levodopa, lipase, lofepramine, lorazepam, loxapine, maprotiline, mazindol, mefenamic acid, melatonin, melitracen, memantine, meperidine, meprobamate, mesalamine, metapramine, metaxalone, methadone, methadone, methamphetamine, methocarbamol, methyldopa, methylphenidate, methylsalicylate, methysergid(e), metoclopramide, mianserin, mifepristone, milnacipran, minaprine, mirtazapine, moclobemide, modafinil (an anti-narcoleptic), molindone, morphine, morphine hydrochloride, nabumetone, nadolol, naproxen, naratriptan, nefazodone, neurontin, nomifensine, nortriptyline, olanzapine, olsalazine, ondansetron, opipramol, orphenadrine, oxaflozane, oxaprazin, oxazepam, oxitriptan, oxycodone, oxymorphone, pancrelipase, parecoxib, paroxetine, pemoline, pentazocine, pepsin, perphenazine, phenacetin, phendimetrazine, phenmetrazine, phenylbutazone, phenytoin, phosphatidylserine, pimozide, pirlindole, piroxicam, pizotifen, pizotyline, pramipexole, prednisolone,Attorney Docket No. 10013-110W01prednisone, pregabalin, propanolol, propizepine, propoxyphene, protriptyline, quazepam, quinupramine, reboxitine, reserpine, risperidone, ritanserin, rivastigmine, rizatriptan, rofecoxib, ropinirole, rotigotine, salsalate, sertraline, sibutramine, sildenafil, sulfasalazine, sulindac, sumatriptan, tacrine, temazepam, tetrabenozine, thiazides, thioridazine, thiothixene, tiapride, tiasipirone, tizanidine, tofenacin, tolmetin, toloxatone, topiramate, tramadol, trazodone, triazolam, trifluoperazine, trimethobenzamide, trimipramine, tropisetron, valdecoxib, valproic acid, venlafaxine, viloxazine, vitamin E, zimeldine, ziprasidone, zolmitriptan, zolpidem, zopiclone and isomers, salts, and combinations thereof.

[0087] In certain embodiments, the exemplary compounds and pharmaceutical compositions can be administered in combination with another antiviral agent(s) such as abacavir, acyclovir, acyclovir, adefovir, amantadine, amprenavir, ampligen, arbidol, atazanavir, atripla, balapiravir, BCX4430, boceprevir, cidofovir, combivir, daclatasvir, darunavir, dasabuvir, delavirdine, didanosine, docosanol, edoxudine, efavirenz, emtricitabine, enfuvirtide, entecavir, famciclovir, favipiravir, fomivirsen, fosamprenavir, foscarnet, fosfonet, ganciclovir, GS-5734, ibacitabine, imunovir, idoxuridine, imiquimod, indinavir, inosine, interferon type III, interferon type II, interferon type I, lamivudine, ledipasvir, lopinavir, loviride, maraviroc, moroxydine, methisazone, nelfinavir, nevirapine, nexavir, NITD008, ombitasvir, oseltamivir, paritaprevir, peginterferon alfa-2a, penciclovir, peramivir, pleconaril, podophyllotoxin, raltegravir, ribavirin, rimantadine, ritonavir, pyramidine, saquinavir, simeprevir, sofosbuvir, stavudine, telaprevir, telbivudine, tenofovir, tenofovir disoproxil, Tenofovir Exalidex, tipranavir, trifluridine, trizivir, tromantadine, truvada, valaciclovir, valganciclovir, vicriviroc, vidarabine, viramidine zalcitabine, zanamivir, or zidovudine and combinations thereof.

[0088] In certain embodiments, pharmaceutical compositions disclosed herein can be administered and / or co-formulated in combination with a second antiviral agent selected from:Attorney Docket No. 10013-110W01

[0089] In certain implementations, the disclosed compounds may be administered in combination with a second antiviral nucleoside disclosed in WO 2016 / 106050, WO 2017 / 156380, WO 2019 / 173602, or WO 2024 / 233354.

[0090] In some implementations, the additional anti-viral agent can be selected from the group consisting of: a compound of this invention, with one or more of ABT-450 and / or ABT-267, and / or ABT- 333, and / or ABT-493, and / or ABT-530; a novel compound of this invention with a compound disclosed in any of US 2010 / 0144608; US 61 / 339,964; US 2011 / 0312973; WO 2009 / 039127; US 2010 / 0317568; 2012 / 151158; US 2012 / 0172290; WO 2012 / 092411; WO 2012 / 087833; WO 2012 / 083170; WO 2009 / 039135; US 2012 / 0115918; WO 2012 / 051361; WO 2012 / 009699; WO 2011 / 156337; US 2011 / 0207699; WO 2010 / 075376; US 7,9105,95; WO 2010 / 120935; WO 2010 / 111437; WO 2010 / 111436; US 2010 / 0168384 or US 2004 / 0167123; a compound of this invention with one or more of Simeprevir, and / or GSK805; a compound of this invention with one or more of asunaprevir, and / or daclastavir, and / or BMS-325; a compound of this invention with one or more of GS-9451, and / or ledisasvir and / or sofosbuvir, and / or GS-9669; a compound of this invention with one or more of ACH- 2684, and / or ACH-3102, and / or ACH-3422; a compound of this invention with one or more of Boceprevir, and / or MK-8742; a compound of this invention with one or more of faldaprevir and / or deleobuvir; a compound of this invention with PPI-668; a compound of this invention with one or more of telaprevir and / or VX-135; a compound of this invention with one or more of samatasvir and / or IDX- 437; a compound of this invention with PSI-7977 and / or PSI-938, a compound of this invention with BMS-790052 and / or BMS-650032; a compound of this invention with GS-5885 and / or GS-9451; a compound of this invention with GS-5885, GS-9190 and / or GS-9451; a compound of this invention in combination with BI-201335 and / or BI-27127; a compound of this invention in combination with telaprevir and / or VX-222; a compound of this invention combination with PSI-7977 and / or TMC-435; and a compound of this invention in combination with danoprevir and / or R7128.

[0091] In some implementations, the administered compound is selected from:Attorney Docket No. 10013-110W01or a pharmaceutically acceptable salt thereof.

[0092] In exemplified embodiments, the pharmaceutical composition includes a compound of any one of formulas (l)-(14pp) and a pharmaceutically acceptable excipient.

[0093] In exemplified embodiments, the pharmaceutical composition includes a compound of any one of formulas (l)-(14pp), or any compound species or combination of compound species as disclosed herein, or a pharmaceutically acceptable salt, solvate, or polymorph thereof, and a pharmaceutically acceptable excipient that is a formulation for oral delivery.

[0094] In exemplified embodiments, the pharmaceutical composition includes any one of formulas (1)- (14pp), or any compound species or combination of compound species as disclosed herein, or a pharmaceutically acceptable salt, solvate, or polymorph thereof, and a pharmaceutically acceptable excipient that is a capsule, a tablet, a cachet, a pill, a powder, a granule, an elixir, a tincture, a suspension, a syrup, or an emulsion.

[0095] In exemplified embodiments, the pharmaceutical composition includes any one of formulas (1)- ( 14pp), or any compound species or combination of compound species as disclosed herein, or a pharmaceutically acceptable salt, solvate, or polymorph thereof, and a pharmaceutically acceptable excipient that is a formulation for oral delivery and is a solid dosage form.

[0096] In exemplified embodiments, the pharmaceutical composition includes any one of formulas (1)- (14pp), or any compound species or combination of compound species as disclosed herein, or a pharmaceutically acceptable salt, solvate, or polymorph thereof, and a pharmaceutically acceptable excipient that is a formulation for parenteral delivery.

[0097] In exemplified embodiments, the pharmaceutical composition includes any one of formulas (1)- (14pp), or any compound species or combination of compound species as disclosed herein, or a pharmaceutically acceptable salt, solvate, or polymorph thereof, and a pharmaceutically acceptableAttorney Docket No. 10013-110W01excipient that is a formulation for parenteral delivery such as bolus injection or infusion, as well as administration by intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular subarachnoid, intraspinal, epidural and intrasternal injection and infusion.

[0098] In exemplified embodiments, the pharmaceutical composition includes any one of formulas (1)- ( 14pp), or any compound species or combination of compound species as disclosed herein, or a pharmaceutically acceptable salt, solvate, or polymorph thereof, and a pharmaceutically acceptable excipient that is a formulation for parenteral delivery such as subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injections or infusion techniques.

[0099] In exemplified embodiments, the pharmaceutical composition includes any one of formulas (1)- (14pp), or any compound species or combination of compound species as disclosed herein, or a pharmaceutically acceptable salt, solvate, or polymorph thereof, and a pharmaceutically acceptable excipient that is a formulation for pulmonary delivery.

[0100] In exemplified embodiments, the pharmaceutical composition includes any one of formulas (1)- (14pp), or any compound species or combination of compound species as disclosed herein, or a pharmaceutically acceptable salt, solvate, or polymorph thereof, and a pharmaceutically acceptable excipient that is a formulation for pulmonary delivery comprising a propellant.

[0101] In exemplified embodiments, the pharmaceutical composition includes any one of formulas (1)- (14pp), or any compound species or combination of compound species as disclosed herein, or a pharmaceutically acceptable salt, solvate, or polymorph thereof, and a pharmaceutically acceptable excipient that is a formulation for pulmonary delivery comprising a propellant such as compressed air, ethanol, nitrogen, carbon dioxide, nitrous oxide, hydrofluoroalkanes (HFA), 1,1, 1,2, -tetrafluoroethane, 1,1,1,2,3,3,3-heptafluoropropane or combinations thereof.EXAMPLES

[0102] The following examples are for the purpose of illustration of the invention only and are not intended to limit the scope of the present invention in any manner whatsoever.Example 1: Synthesis of intermediate IAttorney Docket No. 10013-110W01

[0103] 2-bromo-6-fluorobenzonitrile (Compound 1, 1519 g, 7.50 mol, 1.0 eq) was charged to a 20-L jacketed reactor followed by 6L of MeCN. The mixture was agitated well with an overhead stirrer. 33 wt% methylamine in absolute ethanol (3040 ml, 24.10 mol, 3.21 eq, 2 vol) was added to mixture and stirred at 60 ± 2 °C. for 24 hours. After completion, the mixture was cooled to 5 °C. The resulting solid was collected by vacuum filtration. The filtrate was concentrated under reduced pressure to a solid mass. All solid materials collected were pooled and charged to the reactor. 9L (6 vol) water was added and agitated for 2 hours. The colorless solid was filtered under vacuum and washed with (3 x 1500 mL, 3 x 1 vol). The solid was dried in vacuum oven at 40 ± 5 °C to get 1.594 kg (99% yield) of intermediate I.1H NMR (400 MHz, CDCl3) δ 7.44 – 7.13 (m, 1H), 6.89 (dd, J = 7.9, 0.9 Hz, 1H), 6.60 (dd, J = 8.6, 0.9 Hz, 1H), 4.83 (s, 1H), 2.94 (s, 3H).Example 2: Synthesis of intermediate IIIntermediate I

[0104] Intermediate I (1.583 kg, 7.5 mol, 1.0 eq,) was charged to a 22 L RBF. B2Pin2(2286.0 g, 9.0 mol, 1.20 eq), KOAc (2208.7 g, 22.5 mol, 3.00 eq) were added to flask followed by the addition of 1,4-dioxane (9.5 L, 6 vol) and Pd(dppf)Cl2(274.5 g, 0.375 mol, 0.05 eq). The reaction mixture agitated using an overhead stirrer and degassed with Nitrogen for 15 minutes. The reaction mixture was stirred at 100 ± 5 °C for 16 hours under nitrogen atmosphere. After completion, the reaction mixture was cooled and filtered through a Celite bed (650 g). The solid was washed with 2.4 L of 1,4-dioxane and collected filtrates were pooled and concentrated under reduced pressure. The residue was transferred to a 22 L RBF using 2:8 EtOAc / n-Heptane (15.8 L). The solution was treated with activated carbon (316.7 g, 0.2 wt) and SiliaMetS DMT (158.3 g, 0.1 wt, 0.65 mmol / g). The mixture was stirred at 40-45 °C for 1 h, cooled to room temperature and filtered through a Celite bed (255 gm). The solid cake was washed with 2:8Attorney Docket No. 10013-110W01EtOAc / n-Heptane (12 L). The filtrate was concentrated under reduced pressure and the crude solid product was taken in a 50 L and treated with n-heptane (17.5 L, 8.2 vol). The mixture was agitated with an overhead stirrer at 75 to 80 °C for 1 hour. The mixture was slowly cooled to 25 °C and continued stirring overnight. Next day the slurry was cooled to 5 °C and filtered under vacuum. The resulting colorless solid was washed with n-heptane (2 x 2 L, 2 x 2 vol) and dried in vacuum oven to get 1.44 kg (74% yield) of intermediate II.1H NMR (400 MHz, CDCl3) 67.41 (dd, J = 8.5, 7.2 Hz, 1H), 7.13 (dd, J = 7.2, 1.1 Hz, 1H), 6.77 (dd, J = 8.5, 1.0 Hz, 1H), 2.94 (s, 3H), 1.39 (s, 12H).Example 3: Synthesis of Intermediate III

[0105] 4-bromo-2,7-naphthyridine (Compound 2, 0.50 kg, 2.39 mol, 1.0 eq), and intermediate II (0.70 kg, 2.63 mol, 1.1 eq) were charged to a 22-L three neck jacketed reactor. 2-Me-THF (10 L, 20 vol) was added to the mixture and agitated using an overhead stirrer. Water (2.5 L, 5 vol) and K3PO4(1.015 kg, 4.78 mol, 2.0 eq) were added in sequence and reactor was purged with nitrogen thrice. After charging with XPhos Pd G3 (101.23 g, 0.05 eq) the reactor was nitrogen purged three more times. The reaction mixture was agitated at 60 ± 2 °C for 22 hours. After completion, the reaction mixture was cooled to 25 °C and filtered under vacuum. The solid was washed with (500 mL, 1 vol) water. The organic layer from the filtrate was separated and dried. Combined solid from filtration and evaporation was treated with CPME (2.5 L) and stirred at room temperature for 2 hours. The solid was collected by filtration and dried to get 485 gm of crude intermediate III. It was treated with SiliaMetS DMT (48.0 g, 0.65 mmol / g) in a mixture of DCM (17.1 L) and MeOH (2 L). The mixture was stirred at 40 °C for an hour and at room temperature overnight. The mixture was filtered through Celite and the solid was washed with the cake with 9:1 DCM / MeOH (3.5 L). The filtrate was evaporated to dryness and the SiliaMetS DMT treatment was repeated until desired Pd levels were achieved.1H NMR (400 MHz, DMSO) δ 9.66 (d, J = 1.1 Hz, 1H), 9.64 (d, J = 0.9 Hz, 1H), 8.76 (d, J = 6.0 Hz, 1H), 8.73 (s, 1H), 7.67 - 7.53 (m, 2H), 7.48 (dt, J = 5.9, 1.0 Hz, 1H), 6.89 (dd, J = 8.7, 0.9 Hz, 1H), 6.74 (dd, J = 7 A, 0.9 Hz, 1H), 6.41 (q, J = 4.7 Hz, 1H), 2.86 (d, J = 4.8 Hz, 3H).Attorney Docket No. 10013-110W01Example 4: Synthesis of EIDD-360850 wt% H2O2EtOH-DMSO (1:1)| Intermediate III | EIDD-3608

[0106] Intermediate III (202.6 g, 778.4 mmol, 1.0 eq,) was charged to a 20 L reactor. DMSO (10 vol, 2.02 L) and water (10 vol, 2.02 L) were added to the reactor followed by the addition of 1 N NaOH (778.4 mL, 1.00 eq). The mixture was agitated using an overhead stirrer. After cooling to 5 °C, 50% H2O2(106.7 g, 2.00 eq) was added slowly over a period of 35 minutes, maintaining the temperature below 10 °C. The reaction mixture was stirred at room temperature. Next day another lot of 50% H2O2(36.59 g) was added to the 20-L reactor over 15 minutes, maintain the batch temperature below 10 °C. After stirring for another 16 hours, water (30 vol, 6.08 L) was added to the 20-L reactor over 1.5 h, maintain the batch temperature below 20 °C and stirred for 2 hours at room temperature. Solid was collected by filtration and washed with water (2 x 405 mL, 2 x 2 vol), DCM (405 mL, 2 vol). The crude solid was charged to a IL RBF after drying to which DMF (3 vol, 548 mL) was added and heated 70-75 °C for 2 h. The mixture was slowly cooled and maintained at 0-5 °C, for 2 hours. The solid was filtered and washed with cold methanol (270 mL). The colorless solid was collected and dried in oven at 45-50 °C to get EIDD-3608 as colorless solid 163 gm (75% yield).1H NMR (400 MHz, DMSO-d6) δ 9.56 (d, 7 = 1.0 Hz, 1H), 9.51 (d, J = 0.9 Hz, 1H), 8.77 - 8.48 (m, 2H), 7.53 (d, 7 = 5.9 Hz, 1H), 7.40 - 7.29 (m, 2H), 7.11 (s, 1H), 6.76 (d, 7 = 7.4 Hz, 1H), 6.57 (d, 7 = 7.5 Hz, 1H), 5.47 (q,7 = 4.9 Hz, 1H), 2.81 (d, 7 = 5.0 Hz, 3H).Example 5: Synthesis of EIDD-3426:

[0107] EIDD-3426 and the remaining compounds of the examples were prepared analogously to EIDD- 3608.1H NMR (400 MHz, DMSO) δ 9.48 (s, 1H), 8.71 (d, J = 5.6 Hz, 1H), 8.61 (d, 7 = 6.0 Hz, 1H), 8.02 (d, 7= 5.6 Hz, 1H), 7.95 (s, 1H), 7.65 - 7.55 (m, 2H), 7.52 - 7.39 (m, 2H), 7.31 (d, 7 = 7.5 Hz, 1H);19F NMR (376 MHz, DMSO) 6 -114.47 (dd, 7 = 9.9, 5.7 Hz);13C NMR (101 MHz, DMSO) 6 165.26, 160.25, 158.72 (d, 7 =Attorney Docket No. 10013-110W012.6 Hz), 157.79, 152.58, 144.97, 143.84, 138.84 (d, J = 3.8 Hz), 130.91 (d, J = 8.8 Hz), 130.60, 128.68, 127.05 (d, J = 18.9 Hz), 126.85 (d, 7 = 3.0 Hz), 119.33 (d, 7 = 13.3 Hz), 116.68 (d, 7 = 22.5 Hz); MS (ES-API) [M+l]+: 268.0.)Example 6: Synthesis of EIDD-3452:O

[0108] 1H NMR (400 MHz, DMSO) δ 9.60 (s, 1H), 9.57 (s, 1H), 8.71 (d, J = 6.0 Hz, 1H), 8.67 (s, 1H), 7.94 (s,2H), 7.62 (ddd, 7 = 8.4, 7.6, 5.9 Hz, 2H), 7.53 - 7.42 (m, 3H), 7.27 (dd, 7 = 7.6, 1.0 Hz, 1H);19F NMR (376 MHz, DMSO) 6 -115.81 (dd, 7 = 9.4, 5.9 Hz);13C NMR (101 MHz, DMSO) 6 165.53, 158.84 (d, 7 = 246.0 Hz), 153.47, 153.24, 147.58, 146.57, 137.02, 135.10 (d, 7 = 4.4 Hz), 130.81 (d, 7 = 8.8 Hz), 129.47 (d, 7 = 2.2 Hz), 127.88 (d, 7 = 19.7 Hz), 127.50 (d, 7 = 3.1 Hz), 123.24, 117.90, 116.24 (d, 7 = 22.0 Hz); MS (ES-API) [M+l]+: 268.0.Example 7: Synthesis of EIDD-3482:N\ '

[0109] 1H NMR (400 MHz, DMSO) δ 9.46 (s, 1H), 9.11 (d, J = 4.3 Hz, 1H), 8.58 (d, 7 = 5.8 Hz, 1H), 8.00 (s, 1H), 7.69 (d, 7 = 4.4 Hz, 1H), 7.63 (ddd, 7 = 8.4, 7.6, 5.8 Hz, 1H), 7.54 - 7.45 (m, 3H), 7.28 (dd, 7 = 7.6, 1.0 Hz, 1H);19F NMR (376 MHz, DMSO) 6 -115.43 (dd, 7 = 9.0, 5.9 Hz);13C NMR (101 MHz, DMSO) 6 165.29, 158.79 (d, 7 = 246.3 Hz), 153.94, 152.11, 144.86, 144.20, 143.15, 135.94 (d, 7 = 4.3 Hz), 131.02 (d, 7 = 8.8 Hz), 130.07, 127.13 (d, 7 = 20.1 Hz), 126.73 (d, 7 = 3.0 Hz), 125.85, 118.89, 116.62 (d, 7 = 22.1 Hz); MS (ES- API) [M+l]+: 268.0.Example 8: Synthesis of EIDD-3545:Attorney Docket No. 10013-110W01

[0110] 1H NMR (400 MHz, DMSO) δ 9.48 (s, 1H), 9.05 (d, J = 4.4 Hz, 1H), 8.58 (s, 1H), 7.74 (s, 1H), 7.60 (d, J = 4.5 Hz, 2H), 7.46 (s, 1H), 7.15 (d, J = 7.9 Hz, 1H), 6.88 (d, J = 8.0 Hz, 1H), 6.21 (s, 2H);13C NMR (101 MHz, DMSO) 6 165.37, 153.75, 152.24, 148.38, 146.76, 145.61, 143.83, 130.56, 128.21,125.56, 124.65, 120.00, 109.44, 102.47; MS (ES-API) [M+l]+: 294.10.Example 9: Synthesis of EIDD-3546:

[0111] 1H NMR (400 MHz, DMSO) δ 9.55 (d, J = 1.0 Hz, 1H), 9.49 (d, J = 1.0 Hz, 1H), 8.67 (d, J = 6.0 Hz, 1H), 8.56 (s, 1H), 7.72 (s, 1H), 7.52 (d, 7 = 5.9 Hz, 1H), 7.39 (s, 1H), 7.12 (d, J = 7.9 Hz, 1H), 6.86 (d, 7 = 7.9 Hz, 1H), 6.18 (d, 7 = 4.0 Hz, 2H);13C NMR (101 MHz, DMSO) 6 165.65, 153.41, 152.43, 148.07, 147.27, 146.38, 145.44, 137.45, 130.94, 127.08, 125.12, 123.26, 120.81, 117.92, 109.28, 102.35; MS (ES-API) [M+l]+: 294.10.Example 10: Synthesis of EIDD-3569:

[0112] 1H NMR (400 MHz, DMSO) δ 9.42 (s, 1H), 9.04 (d, J = 4.4 Hz, 1H), 8.53 (s, 1H), 7.88 (s, 1H), 7.58 (d, J = 4.4 Hz, 1H), 7.46 (d, J = 5.7 Hz, 1H), 7.34 (d, J = 8.4 Hz, 1H), 7.28 (d, J = 2.6 Hz, 1H), 7.26 (s, 1H), 7.20 (dd, J = 8.4, 2.7 Hz, 1H), 3.90 (s, 3H);13C NMR (101 MHz, DMSO) δ 169.63, 159.69, 153.94, 152.17, 147.40, 143.85, 143.16, 138.88, 132.60, 130.74, 126.98, 125.50, 118.67, 115.79, 114.00, 56.02; MS (ES- API) [M+l]+: 280.20.Attorney Docket No. 10013-110W01Example 11: Synthesis of EIDD-3570:

[0113] 1H NMR (400 MHz, DMSO) δ 9.56 (s, 1H), 9.49 (s, 1H), 8.66 (d, J = 5.9 Hz, 1H), 8.55 (s, 1H), 7.78 (s, 1H), 7.42 (d, J = 5.9 Hz, 1H), 7.34 (d, J = 8.4 Hz, 1H), 7.23 (d, J = 2.6 Hz, 1H), 7.21 - 7.15 (m, 2H), 3.89 (s, 3H);13C NMR (101 MHz, DMSO) 6 169.97, 159.45, 153.44, 152.18, 147.23, 146.28, 139.62, 137.37, 133.16, 131.53, 125.76, 123.31, 117.78, 115.67, 113.86, 55.97; MS (ES-API) [M+l]+: 280.20.Example 12: Synthesis of EIDD-3598:N

[0114] 1H NMR (400 MHz, DMSO) δ 9.45 (s, 1H), 9.08 (d, J = 4.4 Hz, 1H), 8.56 (d, 7 = 5.8 Hz, 1H), 7.71 (s, 1H), 7.69 (d, 7 = 4.4 Hz, 1H), 7.51 - 7.41 (m, 3H), 7.28 (s, 1H), 7.21 - 7.18 (m, 1H), 2.41 (s, 3H);13C NMR (101 MHz, DMSO) δ 170.03, 153.84, 151.98, 146.25, 143.97, 143.22, 139.06, 134.69, 133.09, 130.81, 130.47, 128.46, 127.75, 126.02, 119.31, 19.70; MS (ES-API) [M+l]+: 264.20.Example 13: Synthesis of EIDD-3582:

[0115] 1H NMR (400 MHz, DMSO) δ 9.58 (d, J = 1.1 Hz, 1H), 9.54 (d, J = 1.0 Hz, 1H), 8.69 (d, J = 5.9 Hz, 1H), 8.67 (s, 1H), 7.68 (d, 7 = 1.6 Hz, 1H), 7.48 - 7.38 (m, 3H), 7.23 (d, 7 = 2.4 Hz, 1H), 7.20 (ddd, 7 = 7.0, 1.8, 0.6 Hz, 1H), 2.41 (s, 3 H);13C NMR (101 MHz, DMSO) 6 170.27, 153.34, 152.78, 147.33, 146.75, 139.88, 137.29, 134.55, 132.17, 130.72, 130.48, 128.60, 128.44, 123.30, 118.30, 19.68; MS (ES-API) [M+l]+: 264.10.Example 14: Synthesis of EIDD-3583:Attorney Docket No. 10013-110W01

[0116] NMR (400 MHz, DMSO) δ 9.44 (s, 1H), 9.09 (d, J = 4.3 Hz, 1H), 8.56 (d, J = 5.8 Hz, 1H), 7.75 (d, J = 2.4 Hz, 1H), 7.71 (d, J = 4.4 Hz, 1H), 7.50 (dd, J = 5.7, 1.0 Hz, 1H), 7.44 (t, J = 7.7 Hz, 1H), 7.29 (d, 7 = 2.3 Hz, 1H), 7.15 (dd, 7 = 7.6, 1.1 Hz, 1H), 7.09 (dd, 7 = 7.9, 1.2 Hz, 1H), 2.09 (ddd, 7 = 13.6, 8.4, 5.2 Hz, 1H), 1.01 (dd, 7 = 8.0, 1.8 Hz, 2H), 0.89 - 0.74 (m, 2H); 13C NMR (101 MHz, DMSO) δ 170.06, 153.79, 151.95, 146.23, 143.97, 143.22, 140.56, 139.35, 132.78, 130.50, 128.65, 127.29, 126.11, 124.34, 119.40, 13.24, 9.90, 9.70; MS (ES-API) [M+l]+: 290.20.Example 15: Synthesis of EIDD-3584:

[0117] 1H NMR (400 MHz, DMSO) δ 9.58 (s, 1H), 9.54 (d, J = 1.0 Hz, 1H), 8.68-8.69 (m, 2H), 7.73 (d, 7 = 2.2 Hz, 1H), 7.48 -7.40 (m, 2H), 7.25 (d, 7 = 2.3 Hz, 1H), 7.15 (dd, 7 = 7.6, 1.1 Hz, 1H), 7.07 (dd, 7 = 8.0, 1.1 Hz, 1H), 2.08 (ddd, 7 = 13.7, 8.4, 5.3 Hz, 1H), 1.05 - 0.93 (m, 2H), 0.87 - 0.73 (m, 2H);13C NMR (101 MHz, DMSO) 6 170.30, 153.31, 152.77, 147.31, 146.86, 140.44, 140.17, 137.34, 131.87, 130.70, 128.64, 128.16, 123.98, 123.30, 118.40, 13.23, 9.91, 9.66; MS (ES-API) [M+l]+: 290.20.Example 16: Synthesis of EIDD-3599:Attorney Docket No. 10013-110W01H2N

[0118] 1H NMR (400 MHz, DMSO) δ 9.45 (d, J = 0.9 Hz, 1H), 9.09 (d, 7 = 4.4 Hz, 1H), 8.57 (d, 7 = 5.8 Hz, 1H), 7.81 (d, 7 = 2.2 Hz, 1H), 7.68 (d, 7 = 4.3 Hz, 1H), 7.48 (dd, 7 = 5.7, 1.0 Hz, 1H), 7.44 - 7.40 (m, 1H), 7.39 - 7.35 (m, 1H), 7.28 (ddd, 7 = 8.4, 5.2, 0.6 Hz, 1H), 2.31 (d, 7 = 2.3 Hz, 3H);19F NMR (376 MHz, DMSO) 6 -108.13 - -124.58 (m);13C NMR (101 MHz, DMSO) δ 168.70 (d, 7 = 2.7 Hz), 160.96 (d, 7 = 245.3 Hz), 153.85, 152.00, 145.41, 144.05, 143.21, 141.44 (d, 7 = 3.9 Hz), 130.51, 129.80 (d, 7 = 8.6 Hz), 129.27 (d, 7 = 3.6 Hz), 126.24, 121.86 (d, 7 = 18.2 Hz), 119.18, 115.27 (d, 7 = 23.0 Hz), 12.06 (d, 7 = 4.0 Hz); MS (ES-API) [M+l]+: 282.20.Example 17: Synthesis of EIDD-3600:H2N

[0119] 1H NMR (400 MHz, DMSO) δ 9.59 (d, J = 1.0 Hz, 1H), 9.55 (d, J = 1.0 Hz, 1H), 8.69 (d, J = 6.0 Hz, 1H), 8.65 (s, 1H), 7.78 (d, J = 2.2 Hz, 1H), 7.42 (dt, 7 = 6.0, 1.1 Hz, 1H), 7.40 - 7.32 (m, 2H), 7.27 (ddd, 7 = 8.3, 5.2, 0.6 Hz, 1H), δ 2.30 (d, J = 2.3 Hz, 3H);19F NMR (376 MHz, DMSO) 6 -116.25 - -116.35 (m).13C NMR (101 MHz, DMSO) 6 168.92 (d, 7 = 2.8 Hz), 160.83 (d, 7 = 244.8 Hz), 153.37, 152.99, 147.42, 146.96, 142.18 (d, 7 = 3.8 Hz), 137.39, 130.59 (d, 7 = 8.6 Hz), 129.93, 128.34 (d, 7 = 3.8 Hz), 123.28, 121.65 (d, 7 = 18.1 Hz), 118.18, 115.23 (d, 7 = 23.1 Hz), 12.05 (d, 7 = 4.2 Hz); MS (ES-API) [M+l]+: 282.10.Example 18: Synthesis of EIDD-3628:Attorney Docket No. 10013-110W01H2N

[0120] 1H NMR (400 MHz, MeOD) δ 9.45 (s, 1H), 9.08 (d, J = 4.4 Hz, 1H), 8.55 (s, 1H), 7.73 (d, J = 4.4 Hz, 1H), 7.66 (d, J = 5.8 Hz, 1H), 7.52 (d, J = 8.3 Hz, 1H), 7.29 (d, J = 8.2 Hz, 1H);19F NMR (376 MHz, MeOD) δ -51.07 - -51.79 (2F, m);13C NMR (101 MHz, MeOD) 6 165.05, 152.79, 151.70, 145.84, 144.07, 142.98, 142.76, 141.09, 131.71 (t, J = 255.6 Hz), 130.90, 130.56, 126.56, 125.92, 119.88, 118.72, 110.78; MS (ES- API) [M+l]+: 330.10.Example 19: Synthesis of EIDD-3636:H2N

[0121] 1H NMR (400 MHz, DMSO) δ 9.61 (d, J = 1.1 Hz, 1H), 9.57 (d, J = 0.9 Hz, 1H), 8.71 (d, J = 5.9 Hz, 1H), 8.62 (s, 1H), 8.04 (s, 1H), 7.71 - 7.60 (m, 2H), 7.53 (d, J = 6.0 Hz, 1H), 7.30 (d, J = 8.3 Hz, 1 H);19F NMR (376 MHz, DMSO) δ -42.51 - -52.11;13C NMR (101 MHz, DMSO) δ 164.00, 153.53, 153.25, 147.61, 146.69, 143.20, 140.49, 137.21, 131.72 (t, 7 = 254.1 Hz), 129.76, 129.48, 127.82, 123.19, 122.34, 117.75, 111.07; MS (ES-API) [M+l]+: 330.10.Example 20: Synthesis of EIDD-3660:H2NAttorney Docket No. 10013-110W01

[0122] 1H NMR (400 MHz, DMSO) δ 9.70 (d, J = 1.6 Hz, 1H), 9.18 (dd, J = 1.6, 0.9 Hz, 1H), 8.13 (dt, J = 8.1, 1.1 Hz, 1H), 8.03 (dd, J = 8.1, 7.2 Hz, 1H), 7.87 - 7.75 (m, 2H), 7.35 (d, J = 8.3 Hz, 1H), 7.25 - 7.13 (m, 3H), 3.90 (s, 3H);13C NMR (101 MHz, DMSO) 6 170.01, 159.40, 151.45, 149.38, 139.51, 138.80, 133.76, 133.12, 132.71, 127.25, 126.41, 125.80, 124.92, 115.58, 113.72, 55.97; MS (ES-API) [M+l]+: 280.20. Example 21: Synthesis of EIDD-3661:H2N

[0123] 1H NMR (400 MHz, DMSO) δ 9.69 (d, J = 1.5 Hz, 1H), 9.28 (dd, J = 1.6, 0.9 Hz, 1H), 8.14 (dt, J = 8.2, 1.1 Hz, 1H), 8.03 (dd, 7 = 8.1, 7.3 Hz, 1H), 7.88 (dd, 7 = 7.3, 1.2 Hz, 1H), 7.42 - 7.30 (m, 2H), 7.08 (s, 1H), 6.77 (dd, 7 = 8.4, 1.0 Hz, 1H), 6.57 (dd, 7 = 7.5, 1.0 Hz, 1H), 5.42 (q, 7 = 4.9 Hz, 1H), 2.82 (d, 7 = 5.0 Hz, 3H);13C NMR (101 MHz, DMSO) δ 170.01, 151.34, 149.84, 146.98, 138.95, 134.55, 133.91, 132.55, 129.95, 126.45, 126.09, 124.49, 124.05, 118.56, 110.31, 30.50; MS (ES-API) [M+l]+: 279.20.Example 22: Biological assay

[0124] Data are given in Table 1 below obtained from various in vitro assays. Briefly, test articles were examined in dose-response assays against recombinant Cedar virus (CedV), measles virus (MeV) and human parainfluenzavirus type 3 (HPIV3) expressing nano-luciferase (nanoL) from an additional transcription unit. Target cells (Vero-E6 (CedV, HPIV3) or Vero-CD150 (MeV)) received serial dilutions of test article, followed by infection at a multiplicity of infection (MOI) of 0.05 infectious units / cell and incubation for 36 hours. Luciferase activity was then determined using a BioTek Synergy Hl multimode plate reader, gain setting 135. Active concentrations (EC50, EC90) were calculated based on 4-paramter variable slope regression models. Cytotoxicity (Ptox) was determined through exposure of uninfected cells to serial dilutions of test articles for 72 hrs. Subsequently, PrestoBlue substrate was added and relative cell viability measured based on fluorescence intensity proportional to substrate turnover in metabolically active cells. Toxic (Ptox) concentrations (CC50) were calculated based on 4-paramter variable slope regression models.Table 1: In vitro assay results.Attorney Docket No. 10013-110W01Code Structure Ptox CedV-nanoL MeV-nanoL HPIV3-nanoLI X z N) z z (mM) (mM) (mM) (mM)\ \C=Z QoO= -n= ECS0EC90ECS0ECgo ECso EC90EIDD- >150 4.4 14.4 1.15 2.7 3.6 7.33426 t)C2Mz~z \ / \ / \ / / ==o | QQ QH2NVIEIDD- >150 5.1 10.29 1.5 8.4 0.36 3.4|i J d3452 oEIDD- >150 1.2 2.9 0.45 0.97 0.3 1.83482EIDD- >150 0.59 0.70 0.15 0.70 0.54 0.623545EIDD- >150 0.99 2.54 0.06 0.12 0.31 0.83546 o D\\Ol z za o=='CM X X EIDD- >150 1.69 4.06 1.75 2.0 0.63 0.763569EIDD- >150 4.0 17.28 1.8 2.0 0.18 0.213570EIDD- >150 1.87 11.32 0.56 2.4 1.29 1.973582EIDD- >150 0.44 2.4 0.17 0.4 0.79 1.373583 toH2N jfSAttorney Docket No. 10013-110W01 Code Structure Ptox CedV-nanoL MeV-nanoL HPIV3-nanoLI N) z (mM) (mM) (mM) (mM))o=\ ECS0EC90ECS0ECgo ECso EC90EIDD- >150 1.33 4.0 0.37 1.0 0.97 1.69|l J J3584 o OCz^H2NjfS 0EIDD- >150 0.84 1.86 0.40 0.90 0.62 0.763598EIDD- >150 1.00 2.45 0.44 1.82 0.59 1.273599 toH2NtotoEIDD- >150 2.99 7.67 0.37 2.22 0.81 1.55N J J3600 0H2N]toEIDD- >150 2.15 6.40 0.85 1.40 0.46 1.60II \ \3608* oH2N-totoHN-toU1EIDD- >150 1.5 6.1 0.47 1.9 0.63 1.43628 toH2Nto llcr ytoEIDD- >150 6.2 49 1.8 5.8 0.4 0.9|l J J3636 oH2N-to |loU / toEIDD- totoN116.8 25.6 22.0 5.1 5.6 2.0 5.93660 □ UMH2N^U^Attorney Docket No. 10013-110W01Code Structure Ptox CedV-nanoL MeV-nanoL HPIV3-nanoL(mM) (mM) (mM) (mM)ECS0EC90ECS0EC90ECso EC90EIDD- >150 2.7 >50 1.1 2.0 0.88 2.103661 Cl >H2Nf |l* As noted in the definitions above, compound references " EIDD-3608 / GHP-88310", " EIDD-3608" and " GHP-88310" can be used interchangeably and refer to a compound having a structure given by the formula.Example 23: Broad-Spectrum Polymerase Inhibitor GHP-88310 Blocks Direct-Contact and Airborne Virus Transmission in a Ferret Surrogate Model of Measles

[0125] This study aimed to evaluate the prophylactic and therapeutic effectiveness of GHP-88310 in preventing morbillivirus transmission, utilizing the CDV ferret model as a surrogate for measles virus. Source animals were intranasally inoculated, while contact animals were exposed to the virus through direct contact transmission or airborne transmission. Viral titers in nasal lavage and PBMC samples were measured using TCIDS0titration assays.Cells, Viruses, and Chemicals

[0126] Vero cells (ATCC CCK-81) stably expressing canine signaling lymphocyte activation molecule (Vero cSLAM, VDS) were cultured in Dulbecco's modified Eagle's medium (DMEM) with 7.5% heat- inactivated fetal bovine serum (FBS) and maintained at 37°C (5% CO2). recCDV-5804p was grown and titrated on VDS cells and confirmed by sequencing. GHP-88310 was synthesized and authenticated by Olon USA. Material was provided as dry powder and stored at -20°C until reconstitution. Individual dosing stocks were dissolved in 1% methylcellulose (sonicating water bath, 37°C) directly before administration.CVD Infection of Ferrets

[0127] Female ferrets, aged 3-5 months (purchased from Triple F Farms), were allowed a one-week rest period after arrival before being randomly assigned to groups for all experiments conducted under ABSL-2 conditions. Source ferrets were anesthetized with a dexmedetomidine / ketamine combination and intranasally inoculated with recCDV-5804p in a total volume of 200 pL, with 100 pL administered per nare. Daily nasal lavages were performed using 1 mL of PBS containing 2x antibiotics-antimycotics (Gibco). Twice weekly, ferrets were anesthetized with dexmedetomidine, and blood was collected fromAttorney Docket No. 10013-110W01the cranial vena cava for complete blood cell (CBC) analysis, determination of neutralizing antibodies and PBMC titration. CBCs were analyzed using a VetScan HM5 (Abaxis). All treated ferrets received oral gavage doses of GHP-88309 or GHP-88310, dissolved in 1% methylcellulose, once or twice daily.Direct and Airborne Transmission Conditions

[0128] Direct transmission conditions were established by housing uninfected contact ferrets with a single infected source ferret in a closed single-ventilated cage. Open cage airborne transmission conditions were created by placing three uninfected contact ferrets in an open cage situated in a rack and positioned 35.5cm (front-to-front) from another cage - in a separate rack - containing an infected source ferret. An oscillating fan was placed 35.5cm behind the infected ferret's cage to help direct airflow toward the uninfected ferrets. Closed cage transmission conditions involved placing uninfected contact ferrets in a single-ventilated cage directly connected to another single-ventilated cage housing an infected ferret. The two cages were joined by a 61 cm PVC pipe (10.16 cm diameter), attached using aluminum flanges and covered with perforated steel grating to prevent ferrets from entering the tunnel. Airflow was directed from the infected ferret's cage to the contact ferret's cage.Virus Titration of Nasal Lavage Samples and PBMCs

[0129] Nasal lavage samples were clarified, and the resulting supernatants were immediately titrated using the TCID50 method on VDS cells. PBMCs were isolated from blood via chemical lysis with ACK buffer (150 mM NH4CI, 10 mM KHCO3, 10 pM EDTA, pH 7.4), followed by two washes in PBS and resuspension in DMEM (ref 88310 efficacy paper). Viremia titers were quantified by co-culturing serial dilutions of the extracted PBMCs with VDS cells, and results were reported as TCID50per 10sPBMCs. Neutralizing Antibody Assay

[0130] Plasma samples were heat-inactivated at 56°C for 30 minutes and then clarified by centrifugation at 4,000 x g for 5 minutes. The clarified plasma was serially diluted two-fold in serum-free DMEM, and each dilution was mixed with recCDV-5804p (2xl03TCIDS0units). After a 75-minute incubation at 37°C, the mixtures were transferred to VDS cells seeded in a 96-well plate, with two technical replicates per animal. Neutralization activity was assessed 3 days later by evaluating cytopathic effects via light microscopy.Statistical Analysis and Data Preparation

[0131] Statistical analyses were conducted using two-way ANOVA, followed by appropriate post hoc multiple comparison tests as specified. All analyses were performed using GraphPad Prism (vlO.10). The number of individual biological repeats (n values) and exact P values are indicated in the figure panels. The threshold for statistical significance was set to 0.05.Attorney Docket No. 10013-110W01Ethical Compliance

[0132] All animal procedures were conducted in accordance with the Guide for the Care of Use of Laboratory Animals (NIH) and the Animal Welfare Act Code of Federal Regulations. Ferret studies were approved by the Georgia State University Institutional Animal Care and Use Committee (IACUC) under protocol A22035. All experiments involving infectious agents were approved by the Georgia State University Institutional Biosafety Committee (IBC) and carried out in BSL-2 / ABL-2 containment facilities. Prophylactic GHP-88310 Blocks Direct-Contact Transmission

[0133] Direct contact transmission was first examined, co-housing infected source animals with uninfected sentinels at a ratio of 1:3 per cage for 8 days, from 3 days post infection (dpi) until sources succumbed to the disease (FIG. 1A). Recapitulating prophylactic treatment in a regional outbreak situation, one each of the three contacts per cage received 12 hours before co-housing GHP-88309 at the highest tolerated dose of 50 mg / kg, clinical candidate analog GHP-88310 at a matching dose of 150 mg / kg based on equal plasma exposure, or vehicle volume equivalent. Treatments were administered orally and continued in twice daily (b.i.d.) regimens. All animals were sampled daily for shed virus titers in nasal lavages, complete blood counts (CBC) and viremia titers in the contacts were determined in 3-4 day intervals, and neutralizing antibody (nAb) titers were assessed weekly. Sampling of individual groups was discontinued when nasal lavages and PBMCs were virus-negative for 3 consecutive measurements, animals succumbed, or 35 days after study start.

[0134] Mean survival of source animals was 11 dpi (FIG. IB). Clinical signs emerged first 3 dpi and comprised, in order of appearance, fever, body weight loss, rash, diarrhea, and conjunctivitis (FIGs. BASE). Animals became viremic 3 dpi (FIG. 1C) and started to efficiently shed virus into nasal lavages 7 dpi (FIG. 9D). All vehicle-treated contacts succumbed by study day 19 (FIG. IB), indicating that infection must have occurred on study day 7-8, correlating an upper respiratory CDV load of approximately 103TCIDso units / ml with productive transmission. All contacts in the prophylactic GHP-88310 group remained disease-free during the duration of the study and none became viremic or virus positive in nasal lavages (FIGs. 1C-1D). In contrast, prophylactic GHP-88309 delayed onset, and alleviated severity, of clinical signs, but all animals of this group became viremic, started to shed efficiently into nasal lavages, and the majority ultimately succumbed with a mean survival of 32 days, resembling the structurally unrelated morbillivirus polymerase inhibitor ERDRP-0519. Consistent with complete suppression of viremia by GHP-88310, none of the animals in the group experienced any change in CBC profile, whereas animals treated with GHP-88309 suffered from severe, albeit transient, morbillivirus lymphocytopenia, which matched in intensity that of the vehicle cohort (FIG. IE). Two of three animalsAttorney Docket No. 10013-110W01in the GHP-88310 group and the single survivor in the GHP-88309 group mounted a protective nAb response approximately 3 weeks after co-housing (FIG. IF).Dynamics of Airborne Morbillivirus Transmission

[0135] For long-distance transmission, open air-contact (oAC) and closed air-contact (cAC) settings were explored in comparison (FIG. 2A). The former placed source and contact ferrets in open cages arranged in parallel, with directional room-airflow of 2,460 ft3 / min through stationary fanning. The latter employed closed cages connected through 4" ID air-tubes with barrier grids on either end and directional airflow of XX in3 / min. Distance between cages was 35.5 vs 61 cm, relative humidity and temperature were kept as consistent as technically at 52 vs 56% and 21 vs 22°C, respectively. Contact time was 5-11 dpi with a source to contact ratio of 1:3 in either setting. At the end of the air-exposure period, contact animals were separated into individual ventilated cages and monitored as before until study day 49.

[0136] Mean survival of animals in the cAC cohort was study day 21 (FIG. 2B), indicating that transmission reliably occurred within the first 4-5 days of shared airspace. Viremia and shed virus load profiles were highly consistent across all animals in the group (FIGs. 2C-2D), confirming synchronized transmission under these conditions. In contrast, oAC sentinels succumbed over a broad 2-day period (mean survival study day 30.5) and onset of viremia and virus shedding was highly variable between individual animals over a broad time period (FIGs. 2B-2D), attesting to overall less efficient virus spread in both time to productive infection and inoculum amount transferred.

[0137] Comparison of the onset of major morbillivirus-associated clinical signs after natural airborne transmission in oAC or cAC settings versus artificial intranasal inoculation illuminated a strong correlation between disease dynamics and route of infection (FIG. 2E). In each setting, all contacts were eventually infected and succumbed to the disease. However, artificially inoculated animals developed first clinical signs (fever) 3 dpi, whereas cAC animals showed first signs of disease 5, and oAC animals 9 dpi. All individual animals in the natural infection, cAC, and oAC cohorts presented with at least one disease marker a respective 5, 11, and 28 days after exposure. Based on these results, the cAC setting was selected for subsequent experiments as the best compromise between natural infection and predictable disease timeline.Prophylactic and Therapeutic GHP-88310 Protect Against Airborne Viral Challenge

[0138] In addition to a vehicle-control arm, four GHP-88310 treatment regimen to mitigate airborne transmission were explored (FIG. 3A): prophylactic at 150 mg / kg b.i.d.; therapeutic at 150 mg / kg b.i.d., started 3 days after exposure to simulate post-exposure prophylaxis (PEP) or 6 days after exposureAttorney Docket No. 10013-110W01when first clinical signs emerged; and prophylactic at 100 mg / kg q.d., reducing the daily dose to the previously determined lowest efficacious level after artificial inoculation. At the end of the air-contact periods, sentinels were again transferred into individual ventilated cages to exclude the possibility of delayed secondary spread. All animals in each treatment cohort survived, whereas all vehicle-treated animals succumbed with a mean survival of 21 days (FIG. 3 B). Prior to death, animals in the vehicle cohort displayed major clinical signs, whereas animals of the q.d. group showed only minor, transient disease symptoms and animals in all b.i.d. groups remained disease-free (FIGs. 6A-6E). Lymphocytopenia was severe and rapid in vehicle animals, moderate and transient in the q.d. group, and fully mitigated in all animals receiving b.i.d. regimen (FIG. 3C). Of the b.i.d. -treated animals, only those in the cohort started 6 days after air-exposure developed transient mild viremia (FIG. 3D), but virus remained undetectable in nasal lavages (FIG. 3E). In contrast, q.d. -dosed sentinels became moderately viremic and transiently shed virus. However, peak viremia and shed titers remained 1-2 orders of magnitude below those of the vehicle-treated cohort.

[0139] CDV-neutralizing antibodies (nAbs) were first detected in animals of the q.d. group 10 days after cAC exposure was established, followed by rapid development of a robust neutralizing response (FIG. 3F). In contrast, high-dose b.i.d. treatment of cAC animals with GHP-88310 was sterilizing in the prophylactic and 3 dpc cohorts, and 1 of 3 animals in the 6 dpc group. Confirming nAb titration results, only q.d. and the 26dpc animals with positive titer results survived an intranasal rechallenge with CDV 31 days after study start, whereas animals of the prophylactic and 3 dpc b.i.d. cohorts and the single nAb-negative animal of the 6 dpc b.i.d. group rapidly succumbed to the infection (FIG. 3G), indicating that treatment with GHP-88310 preserved their CDV-immunological naive status.Treatment of Infected Source Animals Suppresses Viral Dissemination

[0140] To assess the effect of treatment on contagiousness of source animals, intranasally infected source animals were treated with GHP-88310 at 150 mg / kg b.i.d., first initiated 4 dpi (FIG. 4A). In this study, sources received a reduced inoculum amount of 2 x 103TCID50 units / animal, which was identified as suitable to extend time-to-death by approximately 3 days (FIG. 7), facilitating an expanded co-housing period, without compromising overall outcome and disease dynamics (FIGs. 8A-8H). Accordingly, cAC conditions were established for 5 days, from 10-15 dpi, when shed virus titers in nasal lavages of vehicle- treated source animals peaked (FIG. 4B) and viremia titers remained high (FIG. 4C).

[0141] All GHP-88310-treated source animals survived the infection (FIG. 4D) and experienced only transient viremia with minimal virus shedding into the upper respiratory tract (FIGs. 4B-4C). In contrast, vehicle source succumbed to CDV 15 dpi, and cACs of the vehicle died on study day 25, demonstratingAttorney Docket No. 10013-110W01efficient airborne transmission. All air contacts of the GHP-88310-treated sources survived, none experienced weight loss or fever (FIGs. 9A-9B), and none developed any clinical sign of morbillivirus disease (FIGs. 9C-9E). Consistent with GHP-88310-mitigated viremia, treated sources experienced only mild, transient lymphocytopenia and their contacts remained lymphocytopenia-free (FIG. 4E), whereas sources of the vehicle group and their sentinels suffered, with equivalent kinetics, a severe loss of circulating lymphocytes. Whereas GH P-88310-treated source animals mounted a robust nAb response after recovery from the primary infection, their untreated sentinels remained immunologically naive to CDV (FIG. 4F), indicating that treatment of the source animals had suppressed any airborne transmission.

[0142] These results demonstrate that treatment initiated early after infection, albeit after first onset of clinical signs, substantially shortens, or even fully eliminates, the time during which a treated source is contagious, which may provide an epidemiological advantage in an outbreak situation in addition to direct therapeutic benefit to a treated patient.Discussion

[0143] MeV was suggested to linger in room air for several hours after an infected person was present.Despite high titer shedding of CDV from infected source ferrets in this study and several days of aircontact, transmission between animals in open cages was inefficient. This may reflect the high air exchange rate (>14x per hour) in our vivarium, which far exceeds that of a typical in-door setting. Once directional airflow was established between source and contact cages, airborne transmission was rapid, complete, and ensuing disease dynamics in individual contact animals highly reproducible.

[0144] Using the transmission model, we discovered that the original lead of the GHP class, GHP-88309, mediated complete survival only when treatment was initiated therapeutically, phenocopying the first- generation lead class ERDRP-0519. Since CDV pathogenicity in ferrets exceeds that of MeV in humans, efficacy of prophylactic treatment of human patients may not be equally compromised. However, the intended use of a direct-acting measles therapeutic involves preemptive treatment of all social contacts of an index case without knowledge of their infection status.

[0145] GHP-88310 provides full protection against morbillivirus disease in both prophylactic and therapeutic applications. Consistent with our mapping of therapeutic time windows for treatment of morbillivirus disease, therapeutically administered GHP-88310 was highly efficacious and mediated complete survival of naturally infected contacts provided treatment was initiated prior to peak viremia. For ERDRP-0519, we proposed that prophylactic treatment created a scenario in which virus replication is pharmacologically too attenuated to trigger a robust host antiviral response, but not fully suppressed,Attorney Docket No. 10013-110W01ultimately paving a path to host demise without ever developing severe clinical signs. By analogy, the same appeared to apply to prophylactic GHP-88309. Accordingly, we hypothesized that increasing the inhibitor dose to sterilizing concentrations should address the problem. Due to limitations in oral bioavailability (ERDRP-0519) or tolerability (GHP-88309), this notion could only be tested experimentally after development of GHP-88310 with greatly improved tolerability. Indeed, 3-fold higher than standard dose of prophylactic GHP-88310 was fully efficacious and animals remained immunologically CDV-naive, confirming that sterilizing conditions had been established and that no adaptive immune response was required for protection against infection. However, all animals survived that received prophylactical GHP-88310 at 100 mg / kg q.d., the same daily dose at which prophylactic GHP-88309 failed. We attribute the superior prophylactic efficacy of GHP-88310 to well-tolerated exposure peaks after dosing at 100 mg / kg with transiently sub-inhibitory concentrations towards the end of the daily dosing cycle, which permitted intermittent restart of virus replication.

[0146] MeV-exposed contacts emerge with protective humoral measles immunity after prophylactic or therapeutic treatment with GHP-88310. When receiving prophylactic GHP-88310 once daily, animals experienced mild viremia and transient lymphocytopenia, and all emerged with robust anti-MeV nAb titers at study end, supporting the notion that GHP-88310 installed an equilibrium between pharmacological virus attenuation and activation of the adaptive antiviral immune response that was not available to GHP-88309. If equally applicable to human hosts, GHP-88310-treated individuals exposed to MeV will emerge with robust anti-MeV immunity after treatment, which can be anticipated to provide long-lasting protection against future infections. Importantly, none of the GH P-88310-treated animals challenged with CDV showed signs of atypically enhanced morbillivirus disease.

[0147] Therapeutic treatment of infected sources shortens the duration of virus spread, potentially providing additional epidemiological benefit. Beyond the immediate therapeutic benefit for an infected individuum, the possible epidemiological impact of antiviral therapy is controversially discussed. During the COVID-19 pandemic, for instance, household transmission rates did not significantly differ independent of whether patients had received molnupiravir, paxlovid, or ensitrelvir. Likewise, the effect of treatment on preventing household transmission of influenza virus is inconclusive. However, the epidemiology of MeV in a population with largely protective humoral immunity is distinct from pandemic viral transmission or seasonal influenza, and earliest clinical signs of measles such as Koplik spots are telling disease markers. Under these conditions, it may be a more realistic goal to pharmacologically accelerate control of a regional measles outbreak through treatment initiated early after appearance of clinical signs.Attorney Docket No. 10013-110W01

[0148] Well-tolerated small-molecule therapeutics such as GHP-88310 are uniquely suited to meet the needs of measles patients. Antiviral drug development for measles therapy largely focuses on two candidate classes, small-molecule therapeutics and monoclonal nAb biologies. The demands of cost- effective production, high shelf stability, and oral are best met by small-molecules such as GHP-88310. Especially the prospect of oral treatment of outpatients should elicit greater patient compliance than can be anticipated with injectable biologies. These advantages of small-molecules can be offset by a low barrier to resistance and escape mutations from GHP class compounds indeed emerged in cell culture. However, all resistant viruses were severely attenuated in animals and had lost lethality. Virus populations isolated from infected animals after prolonged treatment contained no resistance mutations, indicating that an unusually high fitness penalty prevents viral escape from GHP compounds in vivo.Example 24. GHP-83310 Demonstrates Favorable Tolerability and Broad-Spectrum Efficacy in Models of Measles and Respiratory Orthoparamyxovirus DiseasesStudy Design

[0149] Cells, normal human bronchial / tracheal epithelial (NHBE) cells, well-differentiated air liquid interface (ALI) human airway epithelial (HAE) cells, ferrets, cotton rats and mice were used as models to examine the efficacy of GHP-88310 and other GHP-88309 analogs against HPIV3, SeV, and CDV. Viruses were administered through intranasal inoculation and virus load was monitored continuously in nasal lavages (ferrets only), and in respiratory tissues of cotton rats and mice extracted when indicated after infection. Virus titers were determined through TCID50-titration / plaque assays.Cells, Viruses, and Compound Synthesis

[0150] Vero E6, Vero E6 TMPRSS2, Vero cells stably expressing canine signaling lymphocyte activation molecule (Vero c-SLAM, VDS), Vero-hSLAM, and BHK-T7 cells (BHK cells stably expressing T7) were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 7.5% heat-inactivated fetal bovine serum (FBS) at 37°C and 5% CO2. Cells were tested regularly for mycoplasma contamination prior to usage. Recombinant SeV-GFP, rCDV 5804p, HPIV3 (clinical isolate 13J5, HPIV3 / Seattle / USA / 13J5 / 2016; KY629774.1), rPIV3-nano (JS strain), rMeV (Edmonston strain), and rCedar virus (CedV)-nano were propagated on Vero E6 TMPRSS2 or VDS cells using DMEM with 7.5% serum, scraped when necessary, clarified, aliquoted, and stored at -80°C. The generation of all GHP- 88309 resistant recombinant viruses used in these studies has been previously described. Virus stock titers were determined through TCIDso-titration (rSeV, rCDV, rMeV, and rCedV) or plaque assay (PIV3), and stocks were stored in aliquots at -80°C. All virus stocks were sequence confirmed prior to usage.Attorney Docket No. 10013-110W01GHP-88310 and other GHP-88309 analogs were synthesized as described herein, authenticated through elemental analysis, and stored as dry powder. Working aliquots were dissolved in DMSO or 1% methylcellulose for in vitro and in vivo studies, respectively.Table 2. Elemental analysis of selected GHP-88309 analogs.Compound ID N MR Spectroscopy1H NMR (400 MHz, DMSO-d6) δ 9.56 (d, J = 1.0 Hz, 1H), 9.51 (d, J = 0.9 Hz, 1H), 8.77 GHP-88310- 8.48 (m, 2H), 7.53 (d, J = 5.9 Hz, 1H), 7.40 - 7.29 (m, 2H), 7.11 (s, 1H), 6.76 (d, J = (EIDD-3608)7.4 Hz, 1H), 6.57 (d, J = 7.5 Hz, 1H), 5.47 (q, 7 = 4.9 Hz, 1H), 2.81 (d, 7 = 5.0 Hz, 3H).TH NMR (400 MHz, DMSO) 69.42 (s, 1H), 9.04 (d, 7 = 4.4 Hz, 1H), 8.53 (s, 1H), 7.88 (s, 1H), 7.58 (d, 7 = 4.4 Hz, 1H), 7.46 (d, 7 = 5.7 Hz, 1H), 7.34 (d, 7 = 8.4 Hz, 1H), 7.28 GHP-88369 (d, 7 = 2.6 Hz, 1H), 7.26 (s, 1H), 7.20 (dd, 7 = 8.4, 2.7 Hz, 1H), 3.90 (s, 3 H);13C NMR (EIDD-3569) (101 MHz, DMSO) 6169.63, 159.69, 153.94, 152.17, 147.40, 143.85, 143.16,138.88, 132.60, 130.74, 126.98, 125.50, 118.67, 115.79, 114.00, 56.02; MS (ES-API) [M+l]+: 280.20.rH NMR (400 MHz, DMSO) 69.56 (s, 1H), 9.49 (s, 1H), 8.66 (d, 7 = 5.9 Hz, 1H), 8.55 (s, 1H), 7.78 (s, 1H), 7.42 (d, J = 5.9 Hz, 1H), 7.34 (d, 7 = 8.4 Hz, 1H), 7.23 (d, 7 = 2.6 GHP-88370Hz, 1H), 7.21 - 7.15 (m, 2H), 3.89 (s, 3H);13C NMR (101 MHz, DMSO) 8 169.97, (EIDD-3570)159.45, 153.44, 152.18, 147.23, 146.28, 139.62, 137.37, 133.16, 131.53, 125.76, 123.31, 117.78, 115.67, 113.86, 55.97; MS (ES-API) [M+l]+: 280.20.TH NMR (400 MHz, DMSO) 69.58 (d, 7 = 1.1 Hz, 1H), 9.54 (d, 7 = 1.0 Hz, 1H), 8.69 (d, 7 = 5.9 Hz, 1H), 8.67 (s, 1H), 7.68 (d, 7 = 1.6 Hz, 1H), 7.48 - 7.38 (m, 3H), 7.23 (d, 7 = GHP-883822.4 Hz, 1H), 7.20 (ddd, 7 = 7.0, 1.8, 0.6 Hz, 1H), 2.41 (s, 3H);13C NMR (101 MHz, (EIDD-3482)DMSO) 6 170.27, 153.34, 152.78, 147.33, 146.75, 139.88, 137.29, 134.55, 132.17, 130.72, 130.48, 128.60, 128.44, 123.30, 118.30, 19.68; MS (ES-API) [M+l]+: 264.10.XH NMR (400 MHz, DMSO) 69.48 (s, 1H), 9.05 (d, 7 = 4.4 Hz, 1H), 8.58 (s, 1H), 7.74 (s, 1H), 7.60 (d, J = 4.5 Hz, 2H), 7.46 (s, 1H), 7.15 (d, 7 = 7.9 Hz, 1H), 6.88 (d, 7 = 8.0 GHP-88345Hz, 1H), 6.21 (s, 2H);13C NMR (101 MHz, DMSO) 6 165.37, 153.75, 152.24, 148.38, (EIDD-3545)146.76, 145.61, 143.83, 130.56, 128.21,125.56, 124.65, 120.00, 109.44, 102.47; MS (ES-API) [M+l]+: 294.10.XH NMR (400 MHz, DMSO) 69.55 (d, 7 = 1.0 Hz, 1H), 9.49 (d, 7 = 1.0 Hz, 1H), 8.67 (d, 7 = 6.0 Hz, 1H), 8.56 (s, 1H), 7.72 (s, 1H), 7.52 (d, 7 = 5.9 Hz, 1H), 7.39 (s, 1H), 7.12 (d, GHP-88346 7 = 7.9 Hz, 1H), 6.86 (d, 7 = 7.9 Hz, 1H), 6.18 (d, 7 = 4.0 Hz, 2H);13C NMR (101 MHz, (EIDD-3546) DMSO) 6 165.65, 153.41, 152.43, 148.07, 147.27, 146.38, 145.44, 137.45, 130.94,127.08, 125.12, 123.26, 120.81, 117.92, 109.28, 102.35; MS (ES-API) [M+l]+:294.10.Primary Cells

[0151] Undifferentiated cells: Normal human bronchial / tracheal epithelial cells (Lonza CC-2540S, lot number 519670, Caucasian, 42-year-old female donor " F3", unknown cause of death, smoker) wereAttorney Docket No. 10013-110W01amplified and expanded in lifeline cell culture media (LIFELINE Cell Technology) for no more than 3 passages. Cells were seeded in 24-well plates for experiments.

[0152] Differentiated ALI HAE cells (MatTek): EpiAirways (AIR-100; 23-year-old healthy male, Caucasian;grown at air-liquid interface for 13 days) were equilibrated after arrival and further differentiated in hanging-top plates (following manufacturers protocol) for 14-21 days prior to use in experiments. ALI HAE cells were fed every two days 6 ml maintenance media (MatTek), rinsed weekly and before a procedure (MatTek, TEER buffer). Transepithelial / transendothelial electrical resistance (TEER) was measured on selected inserts using an EVOM3 (World Precision Instruments) following manufacturers protocol.Virus Titration

[0153] TCIDso: Samples were serially diluted (10-fold starting at 1:10 initial dilution) in serum-free DMEM. Serial dilutions were added to either VDS (rCDV), Vero hSLAM (rMeV), BHKT7 (rCedV), or Vero E6 TMPRSS2 (rSeV) cells seeded in 96-well plates at 1x105cells / ml, 14 hours before infection. Infected plates were incubated for 72-96 hours at 37°C with 5% CO2, and titers were determined (Reed and Muench).

[0154] Plaque assays: HPIV3 virus samples were serially diluted (10-fold starting at 1:10 initial dilution) in serum-free DMEM. Serial dilutions were added to Vero E6 TMPRSS2 cells seeded in 12-well plates at 2xl05cells per well, 20 hours before infection. Ninety minutes post infection, cells were overlaid with 1 ml of 2x DMEM and avicel (1:1 ratio). Infected plates were incubated for 84 hours at 37°C with 5% CO2. Upon incubation, cells were washed twice with PBS, stained with 1% crystal violet (in 20% EtOH), and plaques were enumerated.

[0155] Dose-Response Antiviral Assays and Cytotoxicity Assessment

[0156] Compound stocks were prepared in dimethyl sulfoxide (DMSO) and diluted in cell culture media (final DMSO concentration of less than 0.1% in all wells). For luciferase-based dose-response assays, cells were seeded in white-walled 96-well plates one day prior to the experiment to achieve 50% confluency. Threefold serial dilutions of compounds were prepared in triplicate using an automated Nimbus liquid handler (Hamilton) and transferred to the cells. Immediately after the addition of compound, cells were infected with virus as previously described. Each plate contained four wells of positive and negative control (media containing 100 |1M cycloheximide or vehicle (DMSO), respectively). Luciferase activities were determined 48 hours after infection using Nano-GLO buffer (Promega) and an Hl Synergy plate reader (Biotek). Normalized luciferase activities were analyzed with the following formula: % inhibition = (Signalsample- Signalmin) / (Signalmax- Signalmin) x 100, and dose-response curvesAttorney Docket No. 10013-110W01were further analyzed by normalized nonlinear regression with variable hill slope to determine EC50and EC90.

[0157] Virus yield reduction assays with CDV and SeV: Cells were seeded in 24-well plates. Once reaching 90% confluency, cells were overlaid with compound dilutions (three-fold dilutions, in triplicate) and cells were infected at an MOI of 0.1. CDV: After 28 hours of incubation, the supernatant was removed, and cells were scraped in 300 pl media, subjected to two freeze-thaw cycles, clarified, and the resulting supernatant was collected for titration. SeV: Supernatant was collected after 52 hours and clarified. Samples were titrated and scored using the TCID50method after 4 days.

[0158] Virus yield reduction assays in undifferentiated NHBE cells: Cells were seeded at 60% confluency in a 24-well plate. Upon reaching 80% confluency, cells were overlaid with compound dilutions (50 pM, three-fold dilutions, in triplicate) and were infected at an MOI of 0.1 with HPIV3 (clinical isolate 13J5). Supernatant was collected after 48 hours and clarified. Samples were titrated by plaque assay on Vero E6 TMPRSS2 cells and quantified after 4 days.

[0159] Virus yield reduction assays in EpiAirways: Inserts were rinsed using DPBS and infected on the apical side with 10,000 PFU of HPIV3 (clinical isolate 13J5) per well in 100 pl PBS. Sixty minutes after infection, virus was aspirated to allow for air-liquid exchange. Compound dilutions (50 pM, three-fold dilutions, in triplicate) were added on the basolateral side at the time of infection and ALI HAE cells were incubated for 48 hours (after 24 hours, the basolateral media was exchanged - same static compound concentration). After 20 hours of initial incubation, the apical surface was washed once to remove accumulated mucus. Apical shed was harvested in 300 pl PBS that had been incubated at 37°Cfor 30 min. Clarified samples were titrated by plaque assay on Vero E6 TMPRSS2 cells and measured after 4 days.

[0160] To assess the impact of the GHP-compound class on cellular metabolism, Vero E6 cells were seeded at 6,000 cells per well in a 96-well plate and incubated at 37°C (5% CO2) following the addition of a 3-fold serial dilution of each compound, starting at 150 pM. After 72 hours of incubation, cells were incubated with 10 pl per well of PrestoBlue (ThermoFisher Scientific) for 1 hour at 37°C and fluorescence was measured using an Hl synergy plate reader (Biotek) with excitation and emission wavelengths set at 560 nm and 590 nm, respectively. Cell viability was expressed as a percentage relative to the media control.

[0161] To determine the effect of GHP-88310 on cellular metabolism in primary cells, three to four ALI HAE cultures were exposed to increasing concentrations of GHP-88310 (25 pM - 200 pM) in maintenance media (MatTek). To assess barrier integrity, TEER was measured after 48 hours ofAttorney Docket No. 10013-110W01incubation at 37°C with 5% CO2and immediately prior to compound addition to the basolateral chamber of a 12-well hanging insert plate (MatTek). The measurements are displayed in Ohm*cm2.Confocal Microscopy

[0162] PIV3- and mock-infected ALI HAE cells were infected and treated with GHP-88310 at either 0.5 pM or 10 pM as above. Forty-eight hours after infection the apical shed was harvested, and the cells were prepared for confocal microscopy. All incubation steps of immunostaining were performed at room temperature and in the absence of light as previously described. Briefly, the inserts were fixed for 40 min with 4% paraformaldehyde-PBS, washed once with PBS, cut, and mounted on a chamber slide (Fisher, cat# 154461PK). Cells were permeabilized with 500 pl of 0.5% Triton X-100 in PBS for 2 hours, then washed, and blocked with 500 pl per insert of 5% BSA in PBS containing 0.05% Tween-20 for 1 hour. Inserts were washed and stained with the primary antibody in an antibody-binding solution (2% BSA-PBS) for 1 hour (PIV3: goat anti HPIV3 HN, Abeam ab28584; TJ: mouse anti ZO-1 BD, Biosciences 610966; Mucin / Goblet cells: mouse anti MUC5AC, ThermoFisher MA5-12175). Cells were washed three times and incubated with a conjugated antibody for 45 min (donkey anti-goat Alexa Fluor 568 ThermoFisher A-11057; goat anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody, Alexa Fluor 488, Invitrogen, A-11029). Cells were washed an additional three times in PBS and then incubated for 5 min with Hoechst 34580. Following three additional washes, cells were mounted with coverslips using a drop of Prolong™ Diamond Antifade Reagent (Thermo Scientific, Cat# P36970) and stored overnight.

[0163] Confocal Imaging was performed using the Zeiss Axio Observer Z.l and an LSM 800 confocal microscope with AiryScan, controlled with the Zeiss Zen 3.1 Blue software (Windows 10). This software as well as Adobe Photoshop (25.9.0) were employed for image analysis.Single-Dose Pharmacokinetic Property Profiling in Mice, Cotton Rats, Ferrets, and Dogs

[0164] Mice: Eight-week-old female Balb / cJ mice (The Jackson Laboratory) were acclimated for at least three days prior to study initiation. Mice were weighed and administered 20 mg / kg of GHP-class compounds by oral gavage in 1% methylcellulose (200 pl total volume). Blood samples (150 pl) were collected retro-orbitally at designated time points post dosing, centrifuged at 2,000 rpm for 5 minutes at 4°C, and the resulting plasma was stored at -80°C.

[0165] Cotton rats: Female cotton rats (Envigo), aged 2-3 months, were rested for at least three days prior to study start. Animals were weighed and dosed via oral gavage with 50 mg / kg of GHP-88309 or GHP-88310, formulated in 1% methylcellulose (500 pl total volume). Blood samples (150 pl) were collected at 30 minutes, and at 1, 2, 3, 4 and 8 hours post dosing. Plasma was processed as describedAttorney Docket No. 10013-110W01above. At 8 hours post-dosing, organs (lung, brain, spleen, heart, kidney, small intestine, large intestine, and liver) were harvested and immediately frozen in liquid nitrogen.

[0166] All animal tissues were homogenized with 70% acetonitrile in water that included internal standards. Animal plasma and tissue concentrations of test articles were measured by a qualified LC / MS / MS method in MRM mode on a Q. TRAP 5500 (Sciex, Santa Clara, CA, USA) instrument.

[0167] Ferrets: Female ferrets (Triple F Farm), aged 6-8 months old, were acclimated for a minimum of three days prior to dosing. Animals were weighed and administered GHP-88309 or GHP-88310 via oral gavage at doses of 150 mg / kg (GHP-88309) and 150 or 500 mg / kg (GHP-88310). Both compounds were prepared in 1% methylcellulose, with a final gavage volume of 2 ml. Blood samples (200 pl) were collected at 0.5, 1, 2, 3, 4, 6, 8, 12, and 24 hours post-dose and processed as described previously.

[0168] Dogs: Male and female beagles, aged 10-12 months old, were weighed and administered GHP- 88310 orally at 50, 150, or 500 mg / kg, formulated in 1% methylcellulose (study conducted by SRI Biosciences™). Blood samples (300 pl) were collected at pre-dose, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours post-dosing for analysis. Clinical observations were conducted at pre-dose, immediately after dosing, 2-4 hours post-dose, and again at 24 hours. The same four animals were used across all dose levels, with a minimum washout period of seven days between each dose escalation.Single-Dose GHP-88309 Tolerability in Ferrets

[0169] Female ferrets (Triple F Farm), between 6-8 months old, were acclimated for at least three days.Ferrets were weighed and dosed with GHP-88309 once via oral gavage at doses ranging from 50- l,000mg / kg. The compound was formulated in 1% methylcellulose and a final gavage volume of 2 ml was administered. Rectal temperature and clinical scores were measured before dosing and at 30 minutes, as well as 1, 2, 3, 4, 5, 6, 8, 12, and 24 hours post-dose.

[0170] Clinical scoring evaluated: i) temperature to monitor hypothermia and ii) neurological status to detect signs of acute drug toxicity. Temperature scoring was assigned as follows: 0, >38°C; 1, 37.5- 37.9°C; 2, 37-37.4°C; 3, <37°C. To assess neurological status, each ferret was individually placed in a single-ventilated cage at the time of evaluation to observe its response to a novel environment.Neurological scoring was partially adapted from the Merck Veterinary Manual and assigned as follows: 0, explores objects, weasel war dance, intact reflexes (pinch test); 1, decreased interest in exploring objects; 2, lethargic, and / or altered gait, and / or decreased reflexes; 3, obtunded, and / or seizures, and / or paralysis in one or more limbs. Upon reaching the predetermined endpoint, a clinical score of 3 in either category, ferrets were euthanized.Multidose Tolerability Studies in Cotton Rats and FerretsAttorney Docket No. 10013-110W01

[0171] Cotton rats: Female cotton rats (Envigo), aged 2-3 months were habituated for at least three days prior to the beginning of the study. Cotton rats were weighed and administered GHP-88310 twice daily by oral gavage at doses of 300, 600 and 1,200 mg / kg. The compound was formulated in 0.5% methylcellulose, 5mM sodium citrate, and 0.25% Tween80, with a final gavage volume of 500 pl. Each animal received a total of 14 doses and was monitored daily for bodyweight and temperature using implantable electronic ID transponders (TP-1000, BMDS). Following the final dose, blood samples (150 pl) were collected at designated time points, centrifuged at 2,000 rpm for 5 minutes at 4°C, and the plasma was stored at -80°C.

[0172] Ferrets: Female ferrets (Triple F Farm), between 6-8 months old, were allowed to rest for at least three days. Ferrets were weighed and dosed with GHP-88309 or GHP-88310 twice daily via oral gavage at doses ranging from 50 - 1,000 mg / kg. The compounds were formulated in 1% methylcellulose, with a final gavage volume of 2 ml. Animals were weighed daily, and body temperature was measured using a rectal probe. Ferrets received a total of 14 doses under a twice-daily dosing regimen or 7 doses under a once-daily regimen. Blood samples were collected on days 0, 2, 4, 5, 6 and 7 post-dosing for complete blood cell (CBC) analysis, and on days 1 or 2 and 7 for blood chemistry analysis. CBC analysis was performed using a Vetscan’ HM5 hematology analyzer (Abaxis), following the manufacturer's protocol. For the 500 mg / kg and 1,000 mg / kg dosing groups, as well as the multi-dose once-daily dosing study, maximum plasma concentration (Cmax) at 5 hours post-dose was measured after doses 1, 2, 4, 6, 8, 10, 12, and 14, or daily (once-daily regimen only). Blood samples (150 pl) were collected for plasma preparation as previously described.In Vivo Infections with SeV, HPIV3, and CDV

[0173] Efficacy studies in mice: Female 129xl / SvJ mice (The Jackson Laboratory), 6 weeks of age, were acclimated for at least three days before being randomly assigned to study groups and housed under ABSL-2 conditions for infections with sequence-validated rSeV. Bodyweight and rectal temperature were measured once daily. For infection, mice were anesthetized with isoflurane and intranasally inoculated with 5x10sTCID50 units / ml of rSeV in a total volume of 50 pl PBS (25 pl per nare). Treatment with compounds of the GHP-class was initiated 24 hours post-infection and administered twice-daily via oral gavage at a dose of 150 mg / kg in 200 pl of 1% methylcellulose, continuing through study day 8. A subset of animals was euthanized either five days post-infection or upon reaching a predetermined endpoint, and tracheas and lungs were collected for analysis.

[0174] Efficacy studies in cotton rats: Female cotton rats (Envigo), 3-4 weeks of age, were allowed to rest for at least three days prior to being randomly assigned to study groups and single-housed underAttorney Docket No. 10013-110W01ABSL-2 conditions for infections with sequence-validated HPIV3 clinical isolate 13J5. Bodyweight and body temperature, using an implanted electronic ID transponder (TP-1000, BMDS), were measured once daily. For infection, cotton rats were anesthetized with isoflurane and intranasally inoculated with lxlO7pfu / ml of HPIV3 in a total volume of 100 pl PBS (50 pl per nare). For therapeutic dosing, animals were anesthetized with isoflurane 12 hours post-infection and administered GHP-88309 or GHP-88310 in 500 pl of 1% methylcellulose via oral gavage, either once or twice-daily, through study day 3.

[0175] Efficacy studies in ferrets: CDV unvaccinated female ferrets (Triple F Farms), 8 months of age, were rested for one week prior to random assignment into study groups and housed in groups of three animals under ABSL-2 condition. Bodyweight and rectal temperature were recorded once daily. Ferrets were anesthetized with dexmedetomidine / ketamine and intranasally inoculated with 2xl05TCID50 units of CDV 5804p in a total volume of 200 pl PBS (100 pl per nare). Daily, nasal lavages were performed using 1 ml of PBS supplemented with 2x antibiotics-antimycotics (Gibco). Peripheral blood mononuclear cells (PBMCs) were collected twice weekly for titration of cell-associated viremia and CBC analysis. On days 17, 21, and 35 post-infection blood samples were collected to assess neutralizing antibody titers. For therapeutic treatments, ferrets were administered GHP-88310 by oral gavage following either a once or twice-daily regimen (50 - 300 mg / kg) in 2 ml of 1% methylcellulose, beginning at 3-days postinfection. Ferrets were euthanized either upon reaching a predetermined study endpoint or at the conclusion of the study on day 35 post-infection.Virus Titration in Tissue Samples (Mice and Cotton Rats) and Nasal Lavages (Ferrets Only)

[0176] Organs were weighed and homogenized in 300 pl PBS using a Bead Blaster 24R (Benchmark) set to three 30 second cycles at 4°C, with one minute rest intervals between cycles. Homogenates and ferret lavage samples were clarified by centrifugation at 13,000 x g for 10 minutes at 4°C, then subsequently titrated. Viral titers were expressed as TCID50 units / plaques per gram input tissue or per ml.Virus Titration of PBMCs (Ferrets Only)

[0177] Blood samples were collected at specified time points twice per week. As previously described, viremia was assessed by isolating PBMCs from 1 ml of whole blood. Red blood cells were lysed using ACK buffer (150 mM NH4CI, 10 mM KHCO3, 10 pM EDTA- pH 7.4) for 7 minutes, followed by two PBS washes and resuspension in DMEM. PBMC titers were determined by co-culturing serial dilutions of purified PBMCs with VDS cells and expressed as TCID50 per 106PBMCs.Neutralizing Antibody Assay (Ferrets Only)Attorney Docket No. 10013-110W01

[0178] Blood samples (300 pl) were collected on days 17 and 21 post-CDV infection (and on day 35 for the ferret CDV QD study only) to obtain plasma, which was subjected to heat inactivation (56°C, 30 minutes) and clearance centrifugation (4,000 x g, 5 minutes). Heat-inactivated and clarified plasma was diluted two-fold in serum free DMEM, with a final volume of 50 pl per well. Fifty microliters of rCDV 5804p (2xl03TCID so units) was added to the plasma dilutions, followed by incubation of the plasmavirus mixture at 37°C for 75 minutes. Subsequently, the mixture was added in duplicate to VDS cells in 96-well plates, incubated for 3 days, and cytopathic effect was assessed by light microscopy.Whole-Genome Sequencing of Virus Stocks and Ferret Nasal Lavage Samples

[0179] Whole genome sequencing (WGS) was performed using metagenomic next -generation sequencing (mNGS) as described previously. Briefly, extracted RNA was treated with a TURBO DNA-free Kit (Thermo Fisher, product #AM1907) to remove DNA. RNA was then reverse transcribed using random hexamers (Thermo Fisher, product #N8080127) and Superscript IV (Thermo Fisher, product #18090010), following the manufacturer's protocol. Double-stranded cDNA synthesis was performed using Sequenase v2.0 (Thermo Fisher, product #70775Z1000UN), and the resulting cDNA was purified using 1.8x AMPure XP magnetic beads (Beckman Coulter, product # A63882).

[0180] Metagenomic viral WGS libraries were created from purified double-stranded cDNA with tagmentation reagents from the Illumina DNA Prep with Enrichment Kit (Illumina, product #20025524), followed by 14 cycles of dual-indexed PCR for SeV and 18 cycles for HPIV3 and CDV isolates. Amplified libraries were cleaned with 0.8x AMPure XP magnetic beads. Viral isolate libraries were not subjected to enrichment and proceeded directly to sequencing. Libraries were sequenced 2xl50bp on NextSeq 2000 (CDV, HPIV3) or NovaSeq 6000 (SeV).

[0181] Due to its high viral titer, the sample " CDV ferret study - BID treatment - 50 mg / kg - ferret 2 - D8pl" was sequenced using metagenomic WGS on NextSeq 2000 with lxlOObp read format.

[0182] Viral WGS was also performed on all ferret nasal lavage samples with detectable PFU titers, except " CDV ferret study - BID treatment - 50 mg / kg - ferret 2 - D8pl". These samples were processed using QIAseq xHYB Microbial Hyb& Lib Kit A (Qiagen, product #334525) and a custom Qiagen hybridization panel containing probes designed from CDV strain 5804 (AY386315.1) containing eGFP reporter (Qiagen, product # 334586). Following the manufacturer's protocol, extracted RNA was depleted of ribosomal RNA and converted to double-stranded cDNA. Double-stranded cDNA was then enzymatically fragmented, end-repaired, indexed, purified with 0.9x and l.lx bead cleanup, and amplified with 14 cycles of PCR, followed by a final bead purification, according to the manufacturer's protocol.Attorney Docket No. 10013-110W01

[0183] Pre-capture sequencing libraries were pooled based on PFU titer, with one to four samples per pool, and incubated with the custom capture panel. After overnight hybridization with biotinylated probes, probe-target hybrids were bound to streptavidin-coated magnetic beads and washed to remove unbound library fragments. The enriched libraries were amplified with 20 cycles of post-hybridization PCR and purified via l.lx bead clean-up. Post-amplification library fragment sizes were estimated with the TapeStation 4200 D1000 (Agilent), and concentrations were measured with Qubit 4 Fluorometer and Qubit dsDNA HS Assay Kit. Libraries were sequenced on Illumina NextSeq 2000 instruments using a lxlOObp read format.

[0184] Data analysis: Raw reads were trimmed and quality filtered using fastp (vO.23.4) with the following parameters: --cut mean quality 20 --cut front --cut tail --length required 20 -- low_complexity_filter — trim_poly_g — trim_poly_x. The consensus sequence of "recCDV_5804p WT" was generated by majority voting with Geneious Prime 2025.0, using the CDV plasmid sequence (AY386315.1 background) containing eGFP reporter as a reference.

[0185] Variants were identified using RAVA workflow with default parameters. References used in RAVA analysis: for "recCDV_5804p WT" we used plasmid map as a reference (AY386315.1 background), for the " HPIV3 clinical isolate 13J5" - HPIV3 / Seattle / USA / 13J5 / 2016 (KY629774.1), for "recSeV_GFP" - Respirovirus muris strain 52 (MH557085.1), and, finally, for CDV ferret nasal lavage samples - "recCDV_5804p WT" consensus sequence. Sample " CDV ferret study - BID treatment - 50 mg / kg - ferret 2 - D8pl" had insufficient coverage (mean coverage of ~49x with some CDS regions having Ox coverage) and was therefore excluded from the RAVA analysis. Data availability: raw sequencing data is publicly available in NCBI BioProject PRJNA1217449.In Silico Docking

[0186] Docking studies were performed with MOE (version 2024.06). A structure of MeV L (Protein Data Bank (PDB) 9dus) was used for the docking studies. After protonation and energy minimization, the triangle matcher method was used for initial compound placement, followed by refinement using an induced-fit protocol to dock GHP-88310 or GHP-88309 in the L structure based on resistance data information. For MeV L, residues Y942, A866, and 11009 were selected to identify the target site for docking and resulting docking poses from each polymerase target were compared. The top scoring docking poses conserved between GHP-88309 and GHP-88310 were chosen for further analysis.Cellular Uptake and Clearance in Undifferentiated NHBE Cells

[0187] Primary normal HBTEC cells (donor " F3") were seeded in 48 well plates and utilized for a wash- in / wash-out study once confluency was achieved.Attorney Docket No. 10013-110W01

[0188] Wash-in study: An 80 pM dilution of GHP-88310 was prepared in pre-warmed lifeline media.Following media removal from the NHBE cells, 0.5 ml of either the 80 pM compound-media mixture or a DMSO-media control was added to the wells. Plates were incubated at 37°C and extracted in triplicate at the following timepoints: 15 min, 30 min, 45 min, lh, 1.5h and 2h. Cells were washed twice with cold DPBS, followed by the addition of 250 pl of 70% methanol / 30% water spiked with the internal standard to each well treated with GHP-88310. The mock-treated blank plate (16 well) was extracted with 250 pl of 70% methanol / 30% water (no internal standard) per well at one-hour post-dosing. All samples mixed by pipetting and transferred to 1.5 ml microcentrifuge tubes. After centrifugation at 16,000 x g for 10 minutes at 4°C, the supernatants were transferred to 2 ml tubes and stored at -80°C until mass spectrometry (MS) analysis.

[0189] Wash-out study: An 80 pM dilution of GHP-88310 was prepared in pre-warmed lifeline media.After media removal from the NHBE cells, 0.5 ml of the 80 pM compound-media mixture was added to the wells. Plates were incubated at 37°C (5% CO2) for one hour, washed twice with 0.5 ml warm DPBS, and compound free pre-warmed lifeline media was added. Plates were further incubated for the following time points: 0 min, 15 min, 30 min, 45 min, lh, 1.5h and 2h post preloading. After incubation at the desired time points, cells were washed twice with 0.5 ml of cold DPBS and extracted and processed as described above. Samples were stored and analyzed as above.Recapitulation of Ferret Plasma PK Profile in Epi Airways

[0190] Air liquid interface HAE cells were exposed basolaterally to dynamic GHP-88310 concentrations reflected ferret plasma levels after either 50 mg / kg oral twice-daily dosing on day 7 or a single oral dose of 20 mg / kg. All ALI HAE cell cultures (n=4 per condition: vehicle, 50 or 20 mg / kg bid, and 50 or 20 mg / kg qd) were infected with lxlO4PFU per well of the HPIV3 clinical isolate 13J5 one hour prior to treatment. Infected cultures underwent either four 12-hour treatment cycles (bid) or two 24-hour treatment cycles (qd). Twenty hours post infection the apical side was rinsed once with 400 pl PBS to reduce mucus accumulation. After 48 hours post-infection, apically shed virus was collected by incubating the cultures with 300 pl of DPBS at 37°C for 30 minutes. Viral titers were then determined by plaque assay on Vero E6 TMPRSS2 cells.Statistical Analysis and Data Preparation

[0191] For statistical analysis of studies comparing more than two study groups, a one-way analysis of variance (ANOVA) or two-way ANOVA with multiple comparison post hoc tests as specified were used to assess statistical difference between samples. An unpaired, two-tailed t-test was used to compare compound concentrations across the analyzed organs. Statistical analyses were performed usingAttorney Docket No. 10013-110W01GraphPad Prism (vlO.l. O). All figures were assembled in GraphPad Prism and Adobe Illustrator / Photoshop. Study schematics were created using Adobe Illustrator, with animal illustrations generated via the vector tool. Objects in figures 5 and 6 were drawn in Adobe Illustrator and adapted from BioRender. The number of individual biological replicates (n values) and exact P values are shown in the figures. The threshold of statistical significance (a) was set to 0.05.Ethical Compliance

[0192] All animal work was performed in compliance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and the Animal Welfare Act Code of Federal Regulations. Experiments involving mice, cotton rats and ferrets were approved by the Georgia State University Institutional Animal Care and Use Committee (IACUC) under protocols A23011, A24022 and A22035, respectively. All experiments using infectious material were approved by the Georgia State University Institutional Biosafety Committee (IBCs) and performed in BSL-2 / ABSL-2 containment facilities, respectively.Results

[0193] Prolonged 17-day treatment of CDV disease in ferrets with GHP-88309 was highly effective at a minimally efficacious oral dose of 50 mg / kg b.i.d. and all animals survived the infection. In mice, multidose administration of GHP-88309 at 150 mg / kg (highest dose tested) b.i.d. was well tolerated without adverse events.Synthetic Studies to Overcome Tolerability Limitations of GHP-88309

[0194] To estimate the therapeutic index (Tl) in non-rodent species, oral dose levels were increased to 150 mg / kg in multi-dose PK studies in search of tolerability limits in ferrets (FIG. 10A). Unexpectedly, ferrets in the 150 mg / kg group reached endpoint within hours of the second dose (FIG. 10B, FIG. 16A- 16B). Complete blood count (CBC) analysis revealed a dramatic drop in platelet counts in animals of this group after compound administration (Fig. SIC). Plasma PK analysis of the 50 mg / kg group on day 7 (after 14 doses) revealed high plasma concentration peaks (Cmax43.5 μM) and an overall exposure of 270 μM × hr (FIG. 10C). A single ascending dose PK study confirmed that a dose level of 50 mg / kg was well tolerated, whereas animals receiving 150 mg / ml showed transient paralysis and drop in body temperature, and ferrets dosed at 500 mg / kg died within 4 hours of dosing (FIGs. 10D-10F).

[0195] A synthetic lead development program was launched with the primary objective to improve tolerability in non-rodent species. Six azaquinoline lead candidates emerged from this exercise, each featuring a substituted heterocyclyl ring (FIG. 10G). None showed appreciable cytotoxicity in cell culture (FIG. 10H) and all maintained antiviral potency in vitro within a <2-fold range against the HPIV3 primaryAttorney Docket No. 10013-110W01indication (FIG. 101) and respective <8-fold and <3-fold range against the morbillivirus (FIG. 10J) and henipavirus (FIG. 10K) secondary indications.

[0196] Single-dose PK prescreening in mice at an oral dose level of 20 mg / kg revealed highest plasma exposure for, in descending order, GHP-88382, GHP-88310, and GHP-88370, all within an acceptable range of >0.8 U. M x hr but mitigating excessively high exposure levels of GHP-88309 (Fig. 1L). Based on PK performance, these three analogs were selected for proof-of-concept efficacy testing against the primary respirovirus indication.GHP-88310 Efficacy Against the Primary Respirovirus Indication

[0197] Using the lethal Sendai virus (SeV) 129Xl / SvJ mouse model as a surrogate for human respirovirus disease, the three developmental candidates were screened in comparison with GHP- 88309, each dosed orally at 150 mg / kg b.i.d., initiated 24 hours after infection (FIG. 11A). All animals receiving GHP-88309 and GHP-88310 survived with fully mitigated clinical signs, whereas GHP-88382 mediated only partial survival and GHP-88370 provided no therapeutic benefit (FIG. 11B, FIGs. 17A-17B). Virus load in the upper (FIG. 11C) and lower (FIG. 11D) respiratory tract of GH P-88310-treated animals was significantly reduced compared to vehicle-treated mice. Effect size was equivalent to that achieved by GHP-88309. In contrast, GHP-88382 reduced viral burden only in the lower respiratory tract and GHP- 88370 had no significant effect on viral load.

[0198] Accordingly, GHP-88310 was selected for a dose-to-failure study against the primary indication in the HPIV3 cotton rat model, measuring virus load in the respiratory tract 3.5 days post-infection (dpi; FIG. 11E). In this model, minimal dose of original hit GHP-88309 that significantly reduced virus load in both trachea and lungs was 300 mg / kg b.i.d. (FIGs. 11F-11G; FIGs. 18A-18B). GHP-88310 was substantially more potent, showing lowest efficacious concentrations of 50 mg / kg b.i.d. and 16.5 mg / kg b.i.d. in the upper and lower respiratory tract, respectively. Despite greater efficacy, a single dose oral PK study in cotton rats (FIG. 11H) confirmed an approximately 4-fold lower plasma exposure of GHP- 88310 versus GHP-88309 (FIG. 111). Corresponding variations in tissue distribution appeared greatest in brain, heart, and kidneys, smallest in lung and intestinal tissues (FIG. 11J). However, none of these differences reached statistical significance.

[0199] A 7-day ascending multi-dose PK study with GHP-88310 culminating in a 2,400 mg / kg daily dose (FIG. 11K) was well tolerated without changes in bodyweight, temperature, or other adverse signs (FIG.11L; FIG. 19A-19B) and showed dose-dependent plasma exposure build-up to nearly 2,000 pM x hr at highest dose (FIG. 11M). At the lowest concentration tested, trough plasma concentrations were above 19 pM, corresponding to 5-times the cell-culture EC90concentration of GHP-88310 against HPIV3. BasedAttorney Docket No. 10013-110W01on these high and sustained exposure levels in vivo, we examined anti-HPIV3 efficacy in a once-daily (q.d.) therapeutic dosing regimen, starting treatment of cotton rats 12 hpi (FIG. 11N). Oral GHP-88310 at 150 mg / kg q.d. or higher significantly reduced virus load in trachea and the lower respiratory tract (FIGs.11O-11P) without changes in body weight or temperature (FIG. 20A-20B).GHP-88310 Tolerability in Non-Rodent Toxicology Species

[0200] Having validated oral efficacy of GHP-88310 against the primary indication in cotton rats, PK properties of the compound in ferrets were explored in a single dose oral PK study (FIG. 12A). A dose level of 500 mg / kg GHP-88310 was well tolerated with dose-proportional plasma exposure, but plasma levels did not reach the extreme concentration peaks seen with GHP-88309 (FIG. 12B). Escalating 7-day ascending multi-dose tolerability studies administering up to 2,000 mg / ml daily dose (FIG. 12C) had no negative effect on bodyweight, body temperature, and CBC analysis (FIG. 21A-21E). All animals survived the full 7-day course without signs of adverse effects, in contrast to reference animals dosed with GHP- 88309 at 150 mg / kg, which again reached predefined endpoint within hours of receiving the second dose (FIG. 12D). Maximal plasma concentrations examined once daily 4 hours after dosing plateaued dose-dependently after administration of four (500 mg / kg) to eight (1,000 mg / kg) doses (Fig. 12E).Overall exposure after 7 days (14 doses) ranged dose-dependently from 336 μM × hr (50 mg / kg b.i.d.) to2,140 μM × hr (500 mg / kg b.i.d.), confirming build-up of drug levels in multi-dose versus single dose administration (FIG. 12F). At a dose of 50 mg / kg b.i.d., total plasma exposure of GHP-88310 outperformed that of GHP-88309 with 336 μM x hours versus 270 μM x hours at study end (FIG. 10C vs FIG. 12F).

[0201] CBC analysis showed no deviation of GH P-88310-treated animals from the normal range over the course of the study (Fig. 12G) and whole blood chemistry panels taken on days 2 and 7 were likewise unremarkable (FIG. 22). In contrast, a reference animal that had received two doses of GHP-88309 at 150 mg / kg showed changes in three of four liver enzymes examined - alkaline phosphatase (ALP), alanine transaminase (ALT), and aspartate aminotransferase (AST) - by more than 750% one day postdosing and prior to death, suggesting GHP-88309-induced hepatotoxicity. Due to rapid onset of GHP- 88309 mortality at this dose level, a comprehensive blood chemistry analysis of all animals in the reference group was not possible.

[0202] Based on greatly improved tolerability of GHP-88310 compared to GHP-88309, the compound was advanced to single ascending dose PK testing in beagle dogs, the primary non-rodent toxicology species used to inform investigational new drug (IND)-enabling safety studies (FIG. 12H). Dogs were dosed orally with GHP-88310 at 50, 150, or 500 mg / kg, each followed by assessment of plasma exposureAttorney Docket No. 10013-110W01over a 24-hour period (FIG. 12H). All dose levels were well tolerated without adverse signs or weight loss of the animals (FIG. 121). Plasma exposure ranged from 135 to 2,150 μM × hr (FIG. 12J), demonstrating good dose-proportionality and exceeding that observed in ferrets. These results demonstrated that GHP-88310 plasma exposure levels exceeding those at which GHP-88309 was lethal in ferrets were well tolerated in both ferrets and dogs (FIG. 12B vs 12J).GHP-88310 Efficacy Against Measles-Like Disease in Ferrets

[0203] Measles has emerged as an urgent secondary indication for GHP class paramyxovirus inhibitors due to growing vaccination hesitancy in many high-income countries and endemic MeV transmission in large geographical regions in sub-Saharan Africa and Southeast Asia. Potent oral efficacy of GHP-88309 against the CDV / ferret surrogate model of human measles has been demonstrated. GHP-88310 and GHP-88309 had virtually identical antiviral activity against both MeV (FIG. 10J) and CDV (FIG. 13A) in cell culture, returning active (EC50concentrations within a 2-fold range of each other.

[0204] In a proof-of-concept oral efficacy study, ferrets were infected with a lethal inoculum of the wild type CDV-5804p strain and initiated treatment at 3 dpi, when PBMC-associated viremia and fever become first detectable in this model (FIG. 13B and FIGs. 23A-23B). Exploring two distinct treatment regimens, animals received GHP-88310 either at 50 and 150 mg / kg b.i.d., or 50 and 100 mg / kg q.d., the latter corresponding to the lowest efficacious daily dose of 100 mg / kg of GHP-8830925and half that dose. Treatment was continued in all groups until 10 dpi (150 mg / kg b.i.d. group) or 21 dpi (50 mg / kg b.i.d. and q.d. groups). All treated animals survived the infection (FIG. 13C) and clinical signs were fully mitigated (FIGs. 23A-23B), whereas all animals in a vehicle-treated reference group succumbed to CDV disease with a median survival of 11 dpi. Cell-associated viremia was first detectable 3 dpi, when therapeutic treatment was initiated, and rapidly progressed in vehicle-treated animals to peak viremia titers of approximately 1.3 x 105TCID50 units per 106PBMCs at 7 dpi (FIG. 13D). Treatment at 50 or 100 mg / kg q.d. or 50 mg / kg b.i.d. lowered peak titers by ~1 order of magnitude and resolved viremia with similar kinetics by 15-16 dpi. Treatment at 150 mg / kg reduced peak viremia by 2 orders of magnitude and was sterilizing at 10 dpi. The effect of treatment on shed virus load in nasal lavages closely mirrored that on viremia titers (FIG. 13E). Virus shedding from animals receiving GHP-88310 at 150 mg / kg b.i.d. peaked 2 orders of magnitude lower than in the vehicle group and shedding from treated animals ceased 6 dpi. Low b.i.d. dose animals and animals of both q.d. groups showed biphasic shedding kinetics, characterized by an initial decline after treatment start followed by a second shedding peak at approximately 10 dpi. Infectious virus was undetectable in all treated animals 17 dpi. Lymphocytopenia, a hallmark of morbillivirus disease, was efficiently mitigated by 50 mg / kg b.i.d. or 100 mg / kg q.d. GHP-Attorney Docket No. 10013-110W0188310 and fully suppressed at the 150 mg / kg b.i.d. dose level (FIG. 13F). Animals in the 50 mg / kg q.d. treatment group experienced vehicle-like lymphocytopenia until 9 dpi. Subsequently, lymphocyte counts rapidly stabilized and returned to normal range 27 dpi. Consistent with transient lymphocytopenia, the emergence of neutralizing antibodies was delayed in animals of the 50 mg / kg q.d. cohort compared to the other treatment groups (FIG. 13G). Whole genome sequencing of treatment- experienced virus populations recovered from animals at XX different times after infection did not detect any mutations in polymerase proteins, indicating a high barrier against viral escape from GHP- 88310 in vivo.Molecular Interactions Between GHP-88310 and the Viral L Protein

[0205] The central cavity of the viral P-L polymerase complex has been identified as target site for GHP- 88309-class inhibitors through photo-affinity crosslinking and reduced susceptibility profiling. Several high-resolution structural models of the MeV polymerase complex were recently solved, setting the stage for extraction of the GHP-88310 docking pose. Activity testing of GHP-88310 against panels of recombinant HPIV3 (FIG. 14A), SeV (FIG. 14B), and MeV (FIG. 14C) viruses harboring mutations mediating escape from GHP-88309 demonstrated, with few exceptions, high consistency of resistance sites, suggesting conserved docking poses of both compounds. We defined substitutions that caused a >5-fold increase in EC5o and / or EC90values as major drivers of reduced viral susceptibility (highlighted in FIGs. 14A-14C). Exceptions were the HPIV3 L-T1010A variant, which remained fully sensitive to GHP- 88310, and SeV L-Y942H, which mediated robust resistance to GHP-88310 but moderate escape from GHP-88309.

[0206] Informed by these conserved mediators of resistance and photoaffinity mapping results for GHP-88309, both compounds were docked in silico into a native structural model of the MeV polymerase (PDB: 9DUS). Top scoring poses consistently placed both inhibitors into the same defined pocket at the interface between the L protein RdRP and PRNTase domains (FIG. 14D and FIGs. 24A-24C). However, distinct predicted docking poses emphasized a high-affinity interaction between the GHP- 88310 diazo quinoline ring and residue A866 of the RdRP domain that was not present in the GHP-88309 pose (FIGs. 25A-25B). Major resistance sites lined the binding pocket, predicting direct contact between the ligand and L-protein residues A866, Y942, and T1010. Additional hot-spot sites were located in close proximity, suggesting that viral escape from either inhibitor is predominantly due to primary, rather than long-range secondary, resistance.GHP-88310 Efficacy in Primary Human Cells and OrganoidsAttorney Docket No. 10013-110W01

[0207] To assess the potential of GHP-88310 for human therapy, efficacy in physiologically relevant primary human airway epithelium cells and well-differentiated human airway epithelium organoids. Wash-in wash-out studies assessed intracellular exposure levels in 2-hour windows in undifferentiated human bronchial tracheal epithelial cells (HBTECs) in, respectively, the presence of 80 pM extracellular GHP-88310 or after removal of extracellular compound (FIG. 26). At wash-in, a maximal intracellular exposure plateau of 105pmol / 106cells was reached in <15 minutes (earliest sampling time point), and was sustained for the duration of the experiment. After removal of extracellular GHP-88310, intracellular drug levels declined by approximately 2 orders of magnitude within the 2-hour monitoring window. These results indicate high plasma-membrane permeability and favorable metabolic stability of the compound.

[0208] Efficacy testing of GHP-88310 against the primary HPIV3 indication in undifferentiated HBTECs revealed sub-micromolar potency and a steep Hill slope, resulting in a fully sterilizing antiviral effect at concentrations of 10 pM and greater (FIG. 15A). After generation of well-differentiated airway epithelium organoids grown at air-liquid interface (FIG. 15B), we exposed cultures to ascending concentrations of GHP-88310 in the basolateral chamber of the transwell system for 48 hours each and measured corresponding transepithelial electrical resistance (TEER) as a well-established marker of cytotoxicity in the system. Electrical resistance remained unchanged at approximately 600 ohmxcm2before and after exposure (FIG. 15C), indicating that GHP-88310 was well-tolerated at the highest physiologically achievable concentrations without disturbing tissue integrity.

[0209] Dose-response assays using static concentrations of basolateral GHP-88310 after apical infection of organoids with 5,000 plaque forming units (pfu) HPIV3 returned nanomolar EC50values with favorable hill slope (EC900.13 μM), based on titers of apically-released progeny virions (FIG. 15D). Sterilizing conditions were reached at 1.85 pM GHP-88310 in the basolateral chamber, roughly equivalent to trough plasma concentrations in the multi-dose ferret PK studies when compound was administered at 50 mg / kg. To verify enhanced antiviral potency of HPIV3 in the organoid models compared to immortalized cell lines, we imaged HPIV3 -infected, fixed, and stained organoid sections after incubation in the presence of 0.5 or 10 pM compound. Titration of progeny virions prior to imaging demonstrated consistent with dose-response assay results that 0.5 jiM basolateral GHP-88310 reduced virus titers by approximately 2 orders of magnitude (FIG. 15E). Anti-ZO-1 staining revealed an unperturbed tight junction network in the presence of both concentrations examined, similar to that seen in mock-treated uninfected cultures (FIG. 15F). Only sporadic viral antigen was detectable after incubation of infected cells in the presence of 0.5 pM GHP-88310, and none after incubation with 10 pM compound, whereasAttorney Docket No. 10013-110W01HPIV3 proteins were abundant in organoids that had received vehicle volume-equivalents. Overlay of viral and beta-actin stains revealed that viral replication was concentrated in ciliated cells (FIGs. 27A- 27B), consistent with previous reports for HPIV3.

[0210] To assess GHP-88310 efficacy in human organoids at physiological, dynamic concentrations, ferret plasma PK curves were recapitulated in the basolateral chambers of human airway epithelium organoids infected with HPIV3. Two dose levels, 20 mg / kg and 50 mg / kg, each represented by b.i.d. and q.d. dosing regimens, were simulated over a 48-hour period each (FIG. 15G). At study end, progeny virus yields released from the apical surface were determined. Validating the finding of high anti-HPIV3 potency of static GHP-88310 in the organoids, both high-dose regimens were sterilizing (FIG. 15H). Also simulating a low-dose b.i.d. treatment fully suppressed production of infectious progeny virions (FIG. 151), although plasma PK profiles called for peak GHP-88310 concentrations of only ~15 μM and no compound was present in the basolateral chamber for 6 hours in every 12-hour interval (FIG. 15H). Even recapitulating a 20 mg / kg q.d. plasma exposure profile in the basolateral chamber, which comprised compound-free incubation periods of 18 hours in every 24-hour interval, greatly inhibited virus replication. Progeny virus yield was statistically significantly reduced by over two orders of magnitude compared to organoid cultures that had been incubated in the presence of basolateral vehicle volume-equivalents (FIG. 151). These results demonstrate that potent antiviral efficacy is achieved at physiological concentrations in disease-relevant human tissue models without cytotoxicity limitations. Discussion

[0211] This study identified the broad-spectrum anti-orthoparamyxovirus clinical candidate GHP-88310, which showed favorable tolerability properties in rodent species and higher mammals including dogs. De-risking GHP-88309 illuminated a sharp tolerability limit in higher mammals at concentrations exceeding 100 mg / kg daily dose. This issue was addressed synthetically through development of GHP- 88310, which was tolerated without adverse effects by ferrets and dogs at very high dose levels that far exceeded the lethal dose of GHP-88309 in ferrets. Differential responses between rodents and nonrodents to drug exposure are not unusual and often attributable to the high metabolic rate of rodents, an expanded rodent cytochrome P450 system that results in fast breakdown of drugs with frequently distinct metabolites, and / or the large liver of rodents, relative to body weight, that is highly resistant to chemical stress. In the case of GHP-88309, plasma exposure was high after oral dosing in both rodents and non-rodents, but only non-rodents experienced tolerability limits. At equal dose levels, GHP-88310 exposure trailed that of GHP-88309. The described ascending dose PK studies demonstrated, however, that improved tolerability of GHP-88310 was not a trivial consequence of lower oral bioavailabilityAttorney Docket No. 10013-110W01compared to GHP-88309, since plasma exposure in ferrets and dogs after high-dose GHP-88310 far exceeded lethal levels of GHP-88309 in ferrets. Toxicity of high-dose GHP-88309 in non-rodents results most likely from accumulation of an unidentified metabolite, which is not produced in rodents due to their distinct CYP450 composition and absent from GHP-88310 breakdown products due to its distinct chemical structure.

[0212] A hallmark of viral escape from GHP-88309 is that resistance emerges readily in cell culture, but all escape variants were apathogenic and no escape mutations appeared in vivo. Susceptibility testing of developmental candidate GHP-88310 against recombinant strains of three viral targets -the primary indication HPIV3, surrogate respirovirus SeV, and secondary indication MeV -that each harbored confirmed GHP-88309 escape mutations revealed overlapping susceptibility profiles. All mutations were located in the viral L protein, lining the interface between the RdRP and PRNTase domains in the central polymerase cavity. However, tolerance to GHP-88310 was, in most cases, less robust than escape from GHP-88309. Conserved susceptibility profiles spotlight a common molecular target site of both compounds, whereas reduced robustness of escape from GHP-88310 suggests altered docking poses, which was substantiated by in silico QSAR models. Importantly, the hallmark feature of GHP-88309 resistance appears to equally apply to GHP-88310, since whole genome sequencing of heavily treatment-experienced virus populations recovered from ferrets did not reveal substitutions at known escape hot-spots or any other polymerase residues. These results indicate a high barrier against viral escape from GHP compounds without concomitant loss in pathogenicity.

[0213] This study focused on improving tolerability in higher mammals without compromising antiviral potency, and GHP-88309 and GHP-88310 indeed have equal activity in cell culture. Therefore, it was not expected to achieve a substantial in vivo potency boost in both rodent and non-rodent host species. Quantitation of drug tissue levels of both compounds in the cotton rats excluded improved lung tissue distribution of GHP-88310 as a possible explanation for this approximately 6-fold increase in antiviral efficacy. Whereas the lowest efficacious dose of GHP-88309 and GHP-88310 in the CDV ferret model was the same (50 mg / kg), plasma exposure of GHP-88310 was much lower than that achieved by GHP- 88309 after equal doses. The conclusion of an in vivo potency boost in both models was further supported by the observation that GHP-88310 was efficacious also in a q.d. regimen, lowering the minimally required efficacious daily dose by 50%. We propose that breakdown of GHP-88310 in both rodents and non-rodents may generate a bioactive, but non-toxic, metabolite that carries part of the antiviral efficacy, whereas GHP-88309 metabolism produces a toxic by-product. Both effects, increasedAttorney Docket No. 10013-110W01tolerability and enhanced antiviral efficacy of GHP-88310, have substantially expanded the therapeutic window of the compound against the primary and secondary indication in premier animal models.

[0214] Ex vivo, potency of GHP-88309 against HPIV3 in established cell lines and primary HBTECs was virtually identical. However, an approximately 5-fold higher antiviral potency in disease-relevant, well- differentiated human airway epithelium organoids was noted versus the undifferentiated HBTECs. This effect may be due to a combination of the diverse cell types present in the organoids, their complex 3D- architecture, and the more natural nutrient concentration gradient compared to simple monolayers. Indeed, confocal microscopy identified, consistent with previous reports, ciliated cells as the primary host cell type for HPIV3 replication in the airway epithelium cultures. To test the correlation between oral dose level and antiviral effect, ferret PK-informed dynamic compound concentrations were recapitulated in the basolateral chamber of the transwell cultures, simulating repeat oral dosing ranging from 20 mg / kg q.d. to 50 mg / kg b.i.d. This approach was previously implemented in the development of molnupiravir, and dosing targets identified in human airway epithelium organoids closely predicted real- world data determined in subsequent clinical trials. Simulating plasma concentrations of a 20 mg / kg q.d. GHP-88310 dosing regimen in the basolateral chamber significantly reduced progeny virus yields by several orders of magnitude, which translates under consideration of established species conversion factors to a highly feasible predicted oral human dose target of 200 mg daily. Corroborating these results, higher dose levels were fully sterilizing.

[0215] Simulation of 20 mg / kg b.i.d. or 20 mg / kg q.d. regimens entailed two 6-hour or one 18-hour incubation window, respectively, in every 24-hour period in which no compound was present in the basolateral chamber. Effective viral inhibition despite prolonged incubation in the absence of inhibitor is likely due to inhibition of both replicase and transcriptase activity of the viral polymerase complex by GHP class compounds. This concerted action may arrest the viral replication cycle in the primary transcription stage, preventing exponential expansion of viral protein expression and suppressing production of immune-modulatory viral V and C proteins that counteract the type I host interferon response, which exposes the virus to host cells in an unmitigated antiviral state. Both the HPIV3 cotton rat and CDV ferret models confirmed that effective virus inhibition by only intermittently present GHP- 88310 was not an artifact of the ex vivo organoid system, since q.d. administration of GHP-88310 at the lowest efficacious dose likewise resulted in either model in prolonged periods with undetectable plasma drug levels. Efficacious orthoparamyxovirus therapy with an NNPI does not depend on continuous drug plasma concentrations above the minimum inhibitory level, illuminating an overproportional inhibitory effect of targeting the mononegavirus polymerase complex.Attorney Docket No. 10013-110W01References for Example 23

[0216] The references cited below are hereby incorporated by reference to disclose and describe the methods or materials in connection with which the publications are cited or to provide background for the present disclosure. Any incorporation by reference of documents below is limited such that no subject matter is incorporated by reference that is contrary to the explicit disclosure herein. In the event of inconsistent usages between this document and those documents so incorporated by reference below, the use in the incorporated references should be considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls.1. Plemper, R. K.; Lamb, R. A., Paramyxoviridae: The Viruses and Their Replication. In Fields Virology, 7 ed.; Knipe, D. M.; Howley, P. M.; Whelan, S., Eds. Wolters Kluwer / Lippincott Williams & Wilkins: Philadelphia, 2020; Vol. 1, pp 504-558.2. Lee, B.; Broder, C. 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[0217] The compositions and methods of the appended claims are not limited in scope by the specific compositions and methods described herein, which are intended as illustrations of a few aspects of theAttorney Docket No. 10013-110W01claims and any compositions and methods that are functionally equivalent are intended to fall within the scope of the claims. Various modifications of the compositions and methods in addition to those shown and described herein are intended to fall within the scope of the appended claims. Further, while only certain representative compositions and method steps disclosed herein are specifically described, other combinations of the compositions and method steps also are intended to fall within the scope of the appended claims, even if not specifically recited. Thus, a combination of steps, elements, components, or constituents may be explicitly mentioned herein or less, however, other combinations of steps, elements, components, and constituents are included, even though not explicitly stated. The term "comprising" and variations thereof as used herein is used synonymously with the term "including" and variations thereof and are open, non-limiting terms. Although the terms "comprising" and "including" have been used herein to describe various embodiments, the terms "consisting essentially of" and "consisting of" can be used in place of "comprising" and "including" to provide for more specific embodiments of the invention and are also disclosed. Other than in the examples, or where otherwise noted, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood at the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, to be construed in light of the number of significant digits and ordinary rounding approaches

Claims

Attorney Docket No. 10013-110W01CLAIMSWhat is claimed is:A compound having the formula:Rdcor a pharmaceutically acceptable salt thereof, whereinX1is N or CRX1; wherein RX1is selected from H, D, halo, COOH, CO2C1-6alkyl, CN, Ci-6alkyl, Ci-6heteroalkyl, C3-ecycloalkyl, or Ci-eheterocyclyl;X2is N or CRx2; wherein Rx2is selected from H, D, halo, COOH, CO2Ci-6alkyl, CN, Ci.6alkyl, Ci.6heteroalkyl, C3.6cycloalkyl, or Ci-6heterocyclyl;X3is N or CRx3; wherein Rx3is selected from H, D, halo, COOH, CO2C1-6alkyl, CN, Ci-ealkyl, C1-6heteroalkyl, C3.6cycloalkyl, or C1-6heterocyclyl;wherein at least one of X1, X2, or X3is N;Y1is N or CRV1; wherein RV1is selected from H, D, halo, COOH, CO2C1-6alkyl, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;Y2is N or CRv2; wherein Rv2is selected from H, D, halo, COOH, CO2Ci.6alkyl, CN, Ci-ealkyl, C1-6heteroalkyl, C3-5cycloalkyl, or Ci-gheterocyclyl;Y3is N or CRv3; wherein Rv3is selected from H, D, halo, COOH, CO2Ci-ealkyl, CN, Ci-galkyl, C1-6heteroalkyl, C3-5cycloalkyl, or Ci-eheterocyclyl;Y4is N or CRv4; wherein Ry4is selected from H, D, halo, COOH, C CO2C1-6alkyl, N, C1-6alkyl, Ci.6heteroalkyl, C3-ecycloalkyl, or Ci-eheterocyclyl;wherein at least one of Y1, Y2, Y3, or Y4is N;Z is C(=O) or SO2,R1is H or Ci-ealkyl;R2is H or C1-6alkyl;Attorney Docket No. 10013-110W01Rais selected from H, D, halo, COOH, CO2Ci.6alkyl, CN, Ci-ealkyl, Ci.6heteroalkyl, C3.6cycloalkyl, or Ci. eheterocyclyl;Rbis selected from H, D, halo, CO2Ci-ealkyl, COOH, CN, Ci-ealkyl, C1-6heteroalkyl, C3-ecycloalkyl, or Ci- eheterocyclyl;Rcis selected from H, D, halo, CO2Ci.6alkyl, COOH, CN, Ci-6a Ikyl, Ci-gheteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl; andRdis selected from H, D, halo, COOH, CO2C1-6alkyl, CN, C1-6alkyl, C1-6heteroalkyl, C3-6cycloalkyl, or C1-6heterocyclyl;wherein any two or more of Rx1, Rx2, Rx3, Rvl, Rv2, Rv3, Rv4, R1, R2, Ra, Rb, Rc, and Rdmay together form a ring.

2. The compound of claim 1, having the formula:RX3Rb_ Rb3. The compound of any preceding claim, wherein one of Y1, Y2, Y3, or Y4is N.

4. The compound of any preceding claim, having the formula:Attorney Docket No. 10013-110W01Ryi5. The compound of any preceding claim, wherein one of X1, X2, and X3is N, and the other two are CH.

6. The compound of any preceding claim, having the formula:Attorney Docket No. 10013-110W017. The compound of any preceding claim, wherein Rv4and Rdtogether form a ring.

8. The compound of any preceding claim, having the formula:Rbwherein Q1is null, 0, S, NRn, wherein Rnis H or C1-3alkyl, C1-3alkylene, or Ci-3heteroalkylene.

9. The compound of any preceding claim, having the formula:Attorney Docket No. 10013-110W0110. The compound of any preceding claim, having the formula:

11. The compound of any preceding claim, wherein R2and Ratogether form a ring.

12. The compound of any preceding claim, having the formula:RbRb> Rb13. The compound of any preceding claim, having the formula:Attorney Docket No. 10013-110W0114. The compound of claim 13, having the formula:

15. The compound of claim 14, having the formula:

16. The compound of claim 13 having the formula:Attorney Docket No. 10013-110W01Ry1Ry1wherein Q is selected from S or O; andwherein Ra10and Ra11are independently selected from C1-6alkyl.

17. The compound of any preceding claim, wherein Rdis H.

18. The compound of any preceding claim, wherein Raand Rbtogether form a ring, or Rband Rctogether form a ring.

19. The compound of any preceding claim, having the formula:wherein R3and R3are independently selected from H, D, halo, C1-3alkyl, C1-3heteroalkyl, or R3and R3together form an oxo, a C3-5cycloalkyl, or a C1-6heterocyclyl; andR4and R4are independently selected from H, D, halo, Ci-3alkyl, C1-3heteroalkyl, or R4and R4together form an oxo, a C3-6cycloalkyl, or a C1-5heterocyclyl.

20. The compound of any preceding claim, wherein R1and R2are each H.

21. The compound of any preceding claim, wherein Rais H, D, C3-5cycloalkyl, C1-6alkyl, halo, or C1-6heteroalkyl.Attorney Docket No. 10013-110W0122. The compound of any preceding claim, wherein Rais H, F, cyclopropyl, cyclobutyl, methyl, ethyl, methoxy, ethoxy, thiomethyl, thioethyl, methylamino, dimethylamino, ethylamino, or diethylamino.

23. The compound of any preceding claim, wherein Rbis H, D, C3-6cycloalkyl, Ci.sal kyl, halo, or Ci- eheteroalkyl.

24. The compound of any preceding claim, wherein Rbis H, F, cyclopropyl, cyclobutyl, methyl, ethyl, methoxy, ethoxy, thiomethyl, thioethyl, methylamino, dimethylamino, ethylamino, or diethylamino.

25. The compound of any preceding claim, wherein at least one of Raand Rbis not H.

26. The compound of any preceding claim, having the formula:Attorney Docket No. 10013-110W01x2xy;Y2 X2X3Y<YxyV3o o *1JURV / i "-■■¥)Ac^alkyl 6c1-3alkyl Ac^alkylAttorney Docket No. 10013-110W01 _3alkyl (S-|_3alkyl i-^alkylX2><Y'Y^Y2 X2V;R1" N- R2N|xC1-3alkyl I 'cyalkylC1-3alkyl C 1.38 Iky IAttorney Docket No. 10013-110W0127. The compound of claim 26, wherein Y2is N, and Y1, Y3, and Y4are CH.

28. The compound of claim 26 or 27, wherein X1is N, and X2and X3are CH.

29. The compound of claim 26 or 27, wherein X2is N, and X1and X3are CH.

30. The compound of claim 26 or 27, wherein X3is N, and X1and X2are CH.

31. The compound of claim 1, wherein the compound is selected fromAttorney Docket No. 10013-110W01or a pharmaceutically acceptable salt thereof.

32. A method of treating or preventing a viral infection in a subject in need thereof, comprising administering to the subject the compound of any one of claims 1-31.

33. A method of treating a viral infection in a subject in need thereof, comprising administering to the subject the compound of any one of claims 1-31.

34. A method of preventing a viral infection in a subject in need thereof, comprising administering to the subject the compound of any one of claims 1-31.

35. The method of any one of claims 32-34, wherein the viral infection is a paramyxovirus infection.

36. The method of any one of claims 32-34, wherein the viral infection is an orthoparamyxovirus subfamily infection.

37. The method of any one of claims 32-34, wherein the viral infection is a paramyxovirus infection of the respirovirus, morbillivirus, or henipavirus genus.

38. The method of any one of claims 32-34, wherein the viral infection is a parainfluenza virus or a terrestrial morbillivirus.

39. The method of any one of claims 32-34, wherein the viral infection is a human parainfluenza virus.

40. The method of any one of claims 32-34, wherein the viral infection is human parainfluenza virus type 1 or human parainfluenza virus type 3.

41. The method of any one of claims 32-34, wherein the viral infection is a measles virus or a canine distemper virus.

42. The method of any one of claims 32-41, wherein compound is administered orally, parenterally, inhalationally, intranasally, or topically (including eye drops).Attorney Docket No. 10013-110W0143. The method of 32-42, wherein compound prevents transmission of a paramyxovirus infection from treated animals to untreated sentinels.

44. A pharmaceutical composition comprising the compound according to any one of claims 1-31.

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