Inhibitors of molluscum contagiosum infections and methods of using same - Patents.com

Abstract

The present invention provides novel compounds, compositions and methods for treating, ameliorating and / or preventing an orthopoxvirus infection in a subject in need thereof. In certain embodiments, the orthopoxvirus infection is caused by Molluscum contagiosum.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 63 / 248,670, filed September 27, 2021, which is incorporated by reference in its entirety herein.

[0002] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with Government support under AI125005 awarded by the National Institutes of Health. The Government has certain rights in the invention.

[0003] Sequence Listing An XML file named "046483-7338WO1_Sequence_Listing.txt", created on September 22, 2022, and consisting of 3.2 kilobytes, is incorporated by reference in its entirety into this specification. [Background technology]

[0004] background Molluscum contagiosum (MC) is a skin disease caused by the poxvirus molluscum contagiosum virus (MCV). MC appears as skin lesions that may last for months to years before resolving. MC lesions occur in children, adults, and immunosuppressed individuals and are strictly confined to the skin. MCV is transmitted by direct skin-to-skin contact, sexual contact, autoinoculation by scratching the lesion, and indirect inoculation from contaminated fomites. Lesions may become painful after treatments intended to reduce transmission. Lesions can also be psychologically distressing, especially if they scar. MC occurs in 2-10% of the world's population and accounts for approximately 1% of all diagnosed skin diseases in the United States, with nearly 5% in children. In immunocompromised individuals, this infectious disease can be both severe and protracted. 5%-18% of HIV patients have MC. Often, severe MC disease in AIDS patients begins to resolve while receiving highly active antiretroviral therapy (HAART). However, cases have been reported in which MC lesions developed soon after initiation of HAART, suggesting that immune reconstitution inflammatory syndrome (IRIS) may be involved in the reemergence of MCV.

[0005] Current treatments for MC usually employ physical therapy or chemical drugs, which have variable efficacy or safety, often fail to completely remove the lesions, and may leave scars. In addition, the broad-spectrum antiviral drug cidofovir (i.e., 1-((3-hydroxy-2-phosphonylmethoxy)propyl)cytosine), a dCMP analogue, has been used effectively as a topical or intravenous medication for MC in immunocompromised patients. However, this drug has side effects, including inflammation, erosion, and pain for topical treatment, and possible nephrotoxicity for systemic application. To date, no antiviral therapeutic has been approved for the specific treatment of MC. The development of such an effective and safe therapy has been largely hampered by the inability of MCV to grow in culture.

[0006] Processivity factors (PFs) are attractive antiviral therapeutic targets. Their function is to tether DNA polymerase (Pol) to the template, allowing synthesis of the elongating strand. PFs are specific for their cognate DNA Pol and are essential for DNA synthesis. All DNA Pols, from phages to humans, function with a single cognate PF. However, poxviruses, including the prototype vaccinia virus (VV) and MCV, are somewhat unusual in that a heterodimer containing the A20 and D4 viral proteins constitutes a functional PF. D4, which can also function as a uracil DNA glycosylase repair enzyme, binds to its PF partner A20 but not to E9 Pol. On the other hand, A20 binds to both E9 and D4, suggesting that it acts in part as a bridge indirectly connecting D4 to E9.

[0007] D4 is also absolutely required for DNA synthesis by both MCV and VV (prototype poxvirus), making it an attractive antiviral target. Notably, MCV D4 (mD4) can be equally substituted for VV D4 (vD4) in in vitro DNA synthesis reactions. This is consistent with mD4 sharing 55% amino acid sequence identity and 82% similarity with VV. Furthermore, the hypothetical 3-D structure of mD4 overlaps with the known crystal structure of vD4.

[0008] Thus, there is a need in the art to identify novel compounds and formulations that can be used to treat and / or prevent MC infections in humans.In certain embodiments, such compounds and compositions should be formulated for topical or intradermal administration.The present invention addresses this need. Summary of the Invention

[0009] overview The present disclosure in one aspect relates to a compound of formula (I): TIFF2024536861000001.tif23128, or a salt, solvate, enantiomer, diastereoisomer, geometric isomer, or tautomer thereof, wherein X, Y, R 3 , and R 4 is defined within the scope of this disclosure.

[0010] In another aspect, the present disclosure provides a pharmaceutical composition comprising at least one compound of formula (I) and a pharma- ceutically acceptable excipient. In certain embodiments, the at least one pharma- ceutically acceptable excipient is selected from the group consisting of water, polyethylene glycol (PEG) 400, PEG 300, propylene glycol (PG), benzyl alcohol, polysorbate 80, diethylene glycol monoethyl ether (DEGEE), isopropyl myristate, ethanol, diisopropyl adipate, lactate C, and the like. 12~15 At least one selected from the group consisting of alkyl, thickening agents, hydroxypropyl cellulose, and PEG 4000.

[0011] In certain embodiments, the pharmaceutical composition is formulated for topical administration, hi certain embodiments, the topical administration comprises a gel or ointment.

[0012] In another aspect, the present disclosure provides a method for treating, ameliorating, and / or preventing an orthopoxvirus infection in a human subject in need thereof. In certain embodiments, the method comprises administering a therapeutically effective amount of at least one pharmaceutical composition of the present disclosure or a compound of formula (II): TIFF2024536861000002.tif26128, wherein X, Y, R 3 , and R 4 is defined within the scope of this disclosure.

[0013] In certain embodiments, the orthopoxvirus infection is caused by a virus selected from the group consisting of molluscum contagiosum virus (MCV), amerpox virus, cowpox virus, mousepox virus, horsepox virus, monkeypox virus, raccoonpox virus, tanapox virus, variola (smallpox) virus, yokapox virus, cervidopox virus (deerpox), avipox virus (fowlpox), capripox virus (goatpox), leporipox virus (myxoma virus), parapox virus (orph virus), suipox virus (swinepox), and yatapox virus (yaba-like disease virus). In certain embodiments, the orthopoxvirus infection is caused by molluscum contagiosum virus (MCV).

[0014] In certain embodiments, the compound or composition is applied to the skin of the subject. [Brief description of the drawings]

[0015] The following detailed description of certain embodiments of the invention will be better understood when read in conjunction with the accompanying drawings. For purposes of illustrating the invention, certain embodiments are shown in the drawings. It is to be understood, however, that the invention is not limited to the precise arrangement and equipment of the embodiments shown in the drawings. [Figure 1] Figure 1A shows the interaction of poxvirus 4 processivity factor with its A20 partner (processivity factor complex) and DNA polymerase and DNA template. Figure 1B shows the requirement of D4 in DNA synthesis. The model also suggests non-limiting relevant interactions of E9, A20, and D4 triads. Poxvirus processivity factor complex (D4 and A20) binds its cognate DNA polymerase (E9) and tethers it to DNA. This allows E9 Pol to synthesize an elongated strand by sequentially incorporating nucleotides. In the absence of the processivity factor complex, the catalytic activity of E9 Pol is insufficient to synthesize an elongated strand. [Diagram 2]FIG. 2A shows a sequence comparison between mD4 (SEQ ID NO:1) and vD4 (SEQ ID NO:2). Identical amino acids are shaded. Missing amino acids are indicated with dashes. The C-terminal residues 167-180 and 191-206 (underlined) of vD4 are important for the interaction with A20. FIG. 2B shows the predicted structure of mD4 superimposed on the crystal structure of vD4. The mD4 structure was generated by homology modeling using SWISS-MODEL. [Diagram 3] Figure 3A includes graphs showing the finding that compound 1 binds to D4 as analyzed by DSF (differential scanning fluorescence). Figure 3B is a graph showing that the negative controls cidofovir and ST-246 (tecovirimat) exhibit no binding to D4 compared to compound 1. SPR data in Figure 3B: top left, dose-dependent binding of compound 1 to D4; top right, calculation of the dissociation constant for binding compound 1 to D4; bottom left and bottom right, negative controls showing that CDV and ST-246 cannot bind to D4. [Figure 4] FIG. 4 contains images related to the Drug Affinity Responsive Target Stability DARTS study, demonstrating that exemplary compound 1 binds to the D4 target protein, as evidenced by protection against proteolysis. [Diagram 5]FIG. 5 shows a processivity factor-dependent DNA synthesis assay. This figure shows the assay used to evaluate the activity of molluscum contaminant D4 processivity protein (mD4) for DNA synthesis. Each reaction contains all the necessary components to allow for continuous DNA synthesis of non-radioactive DNA. In this assay, a 100 nucleotide template contains a biotin moiety at its 5' end and a 15 nucleotide primer annealed to its 3' end. The annealed primer template is attached to a streptavidin-coated well of a 96-well plate. An oligonucleotide primer (15mer) is annealed to the 3' end of the biotin-labeled oligonucleotide template. Addition of DNA Pol, D4 and A20 allows for the incorporation of dNTPs and dig-dUTP, which are recognized by a digoxigenin (DIG) antibody conjugated to HRP for colorimetric quantification (405 nm). In the presence of the D4 antiviral therapeutic compound (not shown), nucleotide incorporation fails and no DNA synthesis occurs, as evidenced by the lack of colorimetric detection. This assay has been described in detail (Lin & Ricciardi, 2000, J. Virol Methods 88:219-225; US Pat. No. 6,204,028, which is incorporated herein by reference in its entirety). [Figure 6] FIG. 6 contains graphs showing that Compound 1 blocks mD4-dependent processive DNA synthesis in vitro in a processivity factor-dependent DNA synthesis assay. [Figure 7] Figure 7 includes graphs showing that Compound 1 blocks mD4 hybrid poxvirus infection (potency expressed as EC50; top image), as well as actual viral plaque reduction (bottom image). Stained tissue culture plates of cells - the open circles on the left plate are infectious centers of cells disrupted in the presence of DMSO carrier (control), while the solid color stain on the right plate represents a plate protected by Compound 1. [Figure 8]Figures 8A-8B include graphs demonstrating that the compounds of the invention specifically bind to D4. Figure 8A includes a graph showing that compound 1 does not block DNA synthesis of herpes simplex virus-1 (HSV-1). Figure 8B includes a graph showing that compound 1 does not block the ability of herpes simplex virus-1 (HSV-1) to infect cells. [Figure 9] Figure 9 contains a chart summarizing the biological activity of selected compounds of the invention. These compounds were found to target mD4 and specifically block mD4-dependent processive DNA synthesis and infection of molluscum contaminant mD4 hybrid virus. Similar results were obtained with vaccinia virus. [Figure 10A] Figures 10A-10B include graphs revealing the protein kinetic characteristics of D4 in terms of tryptophan fluorescence (Figure 10A) and susceptibility to proteolysis (Figure 10B). Figure 10A: Tryptophan fluorescence of D4 and MBP in the presence of DMSO at the indicated concentrations. The inset shows the various spectra of the protein in the presence or absence of the highest concentration of DMSO tested. Figure 10B: Susceptibility to proteolysis. Pronase was prepared as a stock solution of 10 mg / ml and serially diluted. Proteins are shown with their corresponding tags for investigation by Western blotting. His-tagged D4 (~27 kDa) was investigated with anti-His antibody, while MBP fusions of human estrogen receptor beta (ERβ-N; ~54 kDa), MBP (C-terminal His8 introduction; ~44 kDa), and the N-terminal 63 amino acid stretch of A20 (A2063; ~52 kDa) were investigated with anti-MBP antibody. [Figure 10B] See legend to Figure 10A. [Figure 11A]Figures 11A-11C include graphs showing certain compound selection studies. Figure 11A: Evaluation of the ability to inhibit in vitro DNA synthesis at a single dose of 500 μM. Figure 11B: Screening selected compounds for their ability to block VACV infection at a single dose of 50 μM. Figure 11C: Screening compounds that block viral infection (antiviral activity ≧50%) for their ability to block progressive DNA synthesis. [Figure 11B] See legend to Figure 11A. [Figure 11C] See legend to Figure 11A. [Figure 12A] Figures 12A-12D include graphs evaluating compound binding to D4 based on (Figure 12A) DSF, (Figure 12B) SPR, and (Figures 12C-12D) DARTS. Figure 12B: For SPR, the sensor chip NTA was cross-linked to the reference flow cell by MBP and D4 to the active flow cell. Req = response from steady state, shown by dashed line in the sensorgram overlay; CDV = cidofovir; and ST-246 = tecovirimat. Figure 12C: Binding of compound 1 to various proteins measured by DARTS. Compound 1 was incubated with diluted crude lysates expressing the indicated proteins and proteolysis was achieved by addition of pronase at a dilution of 1:37.5 for measurement of MBP, 1:150 for A2063, 1:300 for D4, and 1:2400 for ERβ-N. Figure 12D: Binding of poxvirus drugs to D4 measured by DARTS. Pronase was used at a dilution of 1:300. [Figure 12B] See legend to Figure 12A. [Figure 12C] See legend to Figure 12A. [Figure 12D] See legend to Figure 12A. [Figure 13]Figure 13 shows heat traces to assess the binding of Compound 1 to DNA. Heat traces are shown using random double-stranded DNA (sheared DNA, shown as resolved on a 1% agarose gel) or single-stranded DNA by using d(TC)15mer(*). Heat traces are scaled to allow comparison and uncorrected heats (Q) are shown without further deconvolution. [Figure 14A] 14A-14D show the inhibition of DNA synthesis by compounds 29 (FIG. 14A), 99 (FIG. 14B), 186 (FIG. 14C), and 187 (FIG. 14D) in an in vitro mD4-dependent processive DNA synthesis assay. [Figure 14B] See legend to Figure 14A. [Figure 14C] See legend to Figure 14A. [Figure 14D] See legend to Figure 14A. [Figure 15A] Figures 15A-15D show the inhibition of mD4-VV DNA infection by compounds 29 (Figure 15A), 99 (Figure 15B), 186 (Figure 15C), and 187 (Figure 15D). mD4-VV surrogate virus-infected BSC-1 cells were treated with increasing amounts of each compound, and viral plaques were quantified 24 hours post-infection. As a control, HSV1 plaque formation in the presence of compounds was analyzed 53 hours post-infection on Vero cells. Data represent the mean ± SD of plaque numbers from at least two independent experiments performed in duplicate. [Figure 15B] See legend to Figure 15A. [Figure 15C] See legend to Figure 15A. [Figure 15D] See legend to Figure 15A. [Figure 16] FIG. 16 provides several compounds of the present disclosure and the corresponding biological activities and / or physical properties. [Figure 17] Figures 17A-17B show the average cumulative amount of compound recovered in each sample, including the stratum corneum, epidermis, dermis and receptor medium. Figure 17A: Average cumulative amount of compound 111 released. Figure 17B: Average cumulative amount of compound 99 released. [Figure 18] 18A-18B provide the mean epidermal and dermal skin concentrations observed during in vitro skin permeation studies of compounds 111 (FIG. 18A) and 99 (FIG. 18B). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] Detailed Description The present invention relates, in part, to the unexpected discovery of novel inhibitors of molluscum contagiosum virus (MCV) infection in humans. Molluscum contagiosum virus (MCV) only infects humans. In humans, viral infection is limited to the skin and not systemic. In certain embodiments, all of the inhibitors described herein also block the prototype poxvirus, vaccinia. In another embodiment, other poxviruses, including but not limited to camelpox virus, cowpox virus, ectromelia virus, horsepox virus, monkeypox virus, raccoonpox virus, turkeypox virus, smallpox virus, yokapox virus, deerpox virus, fowlpox virus, myxoma virus, orf virus, swinepox virus, and yaba-like disease virus, can be inhibited by the compounds described herein.

[0017] In certain embodiments, the compound of the invention or any composition comprising same treats, prevents, and / or ameliorates MCV infection when applied to the skin of an infected human. In yet another embodiment, the compound of the invention or any composition comprising same is applied to at least one MCV lesion on the skin of an infected human.

[0018] Poxvirus D4 processivity factors are essential for viral replication. The viral D4 and A20 proteins form a complex that serves to tether the viral polymerase to the template, allowing it to synthesize highly elongated strands of DNA. Processivity factors are compelling drug targets based on the specificity of their cognate DNA polymerases.

[0019] The interaction of compound 1 with the D4 protein was demonstrated by three independent biophysical measurements: DARTS (drug affinity responsive target stability); DSF (differential scanning fluorescence); and SPR (surface plasma resonance; Figures 12A-12D). Collectively, these biophysical studies reveal that binding of D4 by compound 1 leads to protein destabilization, thereby preventing continuous DNA synthesis in vivo and thus preventing poxvirus infection of cells in culture.

[0020] In vitro processivity DNA synthesis assays (Lin & Ricciardi, 2000, J. Virol Methods 88:219-225; U.S. Patent No. 6,204,028) (Figure 5) demonstrated that compound 1 can block molluscum contaminant mD4-dependent processive DNA synthesis (Figure 6). Compound 1 was shown to be able to block poxvirus infection in a standard cell plaque reduction assay (Figure 7). Compound 1 was further shown to exhibit specificity, as it was unable to block herpes simplex virus-1 (HSV-1) processive DNA synthesis (Figure 8A) and also to block HSV-1 infection (Figure 8B). Compound 1 bound to D4 abolishes viral DNA synthesis and viral infection. Additional analogs (compounds 2-4) were synthesized and were shown to be able to block mD4-dependent DNA synthesis inhibition (IC 50 ); Infection (IE 50 ); analyzed for cell proliferation and specificity (inability to block HSV-1).

[0021] definition As used herein, each of the following terms has the meaning associated with it in this section.

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, exemplary methods and materials are described below.

[0023] Generally, the nomenclature used herein and the laboratory procedures in pharmaceutical and organic chemistry are those well known and commonly employed in the art.

[0024] As used herein, the articles "a" and "an" refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0025] As used herein, the term "about" is understood by those of ordinary skill in the art and is subject to some variability depending on the context in which the term is used. As used herein when referring to a measurable value, such as an amount, duration, etc., the term "about" is meant to encompass variances of ±20% or ±10%, in certain alternative embodiments ±5%, in alternative embodiments ±1%, and in further alternative embodiments ±0.1% from the specified value, which are adequate to perform the disclosed methods.

[0026] As used herein, the term "D4" refers to D4 processivity factor. Additionally, as used herein, the term "mD4" refers to molluscum contaminant D4 processivity factor.

[0027] As used herein, a "disease" is a state of health in a subject in which the subject is unable to maintain homeostasis and in which the subject's health continues to deteriorate if the disease is not ameliorated.

[0028] As used herein, a "disorder" in a subject is a health condition in which the subject is able to maintain homeostasis, but in which the subject's health condition is less favorable than it would be in the absence of the disorder. If left untreated, the disorder does not necessarily cause a further deterioration in the subject's health condition.

[0029] As used herein, "ED 50 The term "ED50" or "ED50" refers to the effective dose of a formulation that produces about 50% of its maximum effect in a subject to which the formulation is administered.

[0030] As used herein, an "effective amount," "therapeutically effective amount," or "pharmacologically effective amount" of a compound is an amount of the compound sufficient to provide a beneficial effect to the subject to which the compound is administered.

[0031] "Instructional material," as used herein, includes literature, audio directions, diagrams, or any other medium of expression that can be used to communicate the utility of the inventive compositions and / or compounds in the kit. The kit's instructional material may, for example, be affixed to a container that contains the inventive compounds and / or compositions or may be shipped with a container that contains the compounds and / or compositions.

[0032] As used herein, a "patient" or "subject" may be a human or a non-human mammal or bird. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and murine mammals. In certain other embodiments, the subject is a human.

[0033] As used herein, the term "pharmaceutical composition" or "composition" refers to a mixture of at least one compound useful within the invention and a pharma- ceutical acceptable carrier. The pharmaceutical composition facilitates administration of the compound to a subject.

[0034] As used herein, the term "pharmaceutical acceptable" refers to a substance, such as a carrier or diluent, that does not abrogate the biological activity or properties of a compound useful within the invention and is relatively non-toxic, i.e., the substance may be administered to a subject without causing undesired biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0035] As used herein, the term "pharmaceutical acceptable carrier" refers to a pharmaceutically acceptable substance, composition, or carrier, such as a liquid or solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating material, that is involved in carrying or transporting a compound useful within the scope of the invention in or to a patient so that the compound can perform its intended function. Typically, such constructs are carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the scope of the invention, and not harmful to the patient. Some examples of substances which may serve as pharma- ceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyhydric alcohols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; surfactants; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffers; and other non-toxic, compatible substances employed in pharmaceutical preparations. As used herein, a "pharmaceutically acceptable carrier" is compatible with the activity of the compounds useful within the invention and includes any and all coatings, antibacterial and antifungal agents, absorption delaying agents, and the like that are physiologically acceptable to a subject. Supplementary active compounds may also be incorporated into the compositions. A "pharmaceutically acceptable carrier" may further include pharmaceutically acceptable salts of compounds useful within the invention.Other additional ingredients that may be included in pharmaceutical compositions used in the practice of the invention are known in the art and are described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, Pa.), which is incorporated herein by reference.

[0036] As used herein, the term "pharmaceutically acceptable salts" refers to salts of the administered compound prepared from pharmaceutically acceptable non-toxic acids and bases, including inorganic acids, inorganic bases, organic acids, inorganic bases, solvates, hydrates, and clathrates thereof.

[0037] As used herein, the term "pharmaceutical composition" refers to a mixture of at least one compound useful within the scope of the invention with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners and / or excipients. The pharmaceutical composition facilitates administration of the compound to an organism. Various techniques for administering the compound include, but are not limited to, intravenous, oral, aerosol, parenteral, ocular, pulmonary and topical administration.

[0038] The terms "prevent," "preventing," or "prevention," as used herein, refer to avoiding or delaying the onset of symptoms associated with a disease or condition in a subject who does not develop such symptoms at the time administration of an agent or compound is initiated. Disease, condition, and disorder are used interchangeably herein.

[0039] The term "solvate," as used herein, refers to a compound formed by solvation, which is a process of attraction and association of solute molecules or ions with solvent molecules. When solute molecules or ions are dissolved in a solvent, they diffuse out and are surrounded by the solvent molecules.

[0040] The terms "treat," "treating," or "treatment," as used herein, mean reducing the frequency or severity with which a subject experiences symptoms of a disease or condition by administering an agent or compound to the subject.

[0041] As used herein, the term “alkyl,” by itself or as part of another substituent, means, unless otherwise stated, an alkyl group having the specified number of carbon atoms (i.e., C1 to C6 10 means a straight or branched chain hydrocarbon having from 1 to 10 carbon atoms, including straight, branched, or cyclic substituents. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertbutyl, pentyl, neopentyl, hexyl, and cyclopropylmethyl. (C1-C6) alkyl, such as, but not limited to, ethyl, methyl, isopropyl, isobutyl, n-pentyl, n-hexyl, and cyclopropylmethyl, are most preferred.

[0042] As used herein, the term “alkylene,” by itself or as part of another substituent, unless otherwise stated, refers to an alkyl group having the specified number of carbon atoms (i.e., C1 to C6) with two vacant valences. 10 means a straight or branched chain hydrocarbon group having 1 to 10 carbon atoms, including straight, branched, or cyclic substituents. Examples include methylene, 1,2-ethylene, 1,1-ethylene, 1,1-propylene, 1,2-propylene, and 1,3-propylene.

[0043] As used herein, the term "cycloalkyl" by itself or as part of another substituent means, unless otherwise specified, a cyclic chain hydrocarbon having the specified number of carbon atoms (i.e., C3-C6 means a cyclic group containing a ring group of 3-6 carbon atoms) and includes straight chain, branched chain, or cyclic substituents. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Most preferred are (C3-C6)cycloalkyls, such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0044] As used herein, the term "alkenyl", employed alone or in combination with other terms, means, unless otherwise specified, a stable mono- or di-unsaturated, straight or branched chain hydrocarbon group having the specified number of carbon atoms. Examples include vinyl, propenyl (or allyl), crotyl, isopentenyl, butadienyl, 1,3-pentadienyl, 1,4-pentadienyl, and higher homologs and isomers. An example of a functional group representing an alkene is -CH2-CH=CH2.

[0045] As used herein, the term "alkynyl", employed alone or in combination with another term, means a stable straight or branched chain hydrocarbon group with a carbon-carbon triple bond having the indicated number of carbon atoms, unless otherwise specified. Non-limiting examples include ethynyl and propynyl and higher homologs and isomers. The term "propargyl" refers to a group exemplified by -CH2-C≡CH. The term "homopropargyl" refers to a group exemplified by -CH2CH2-C≡CH. The term "substituted propargyl" refers to a group exemplified by -CR2-C≡CR, where R at each occurrence is independently H, alkyl, substituted alkyl, alkenyl, or substituted alkenyl, provided that at least one R group is not hydrogen. The term "substituted homopropargyl" refers to a group exemplified by -CR2CR2-C≡CR, where R at each occurrence is independently H, alkyl, substituted alkyl, alkenyl, or substituted alkenyl, provided that at least one R group is not hydrogen.

[0046] As used herein, unless otherwise specified, the term "alkenylene," employed alone or in combination with other terms, means a stable mono- or di-unsaturated, straight or branched chain hydrocarbon radical having the specified number of carbon atoms with two vacant valences.

[0047] As used herein, the term "alkynylene," employed alone or in combination with another term, means, unless otherwise specified, a stable straight or branched chain hydrocarbon group having a carbon-carbon triple bond with two vacant valences and the specified number of carbon atoms.

[0048] As used herein, the term "substituted alkyl," "substituted cycloalkyl," "substituted alkenyl," "substituted alkynyl," "substituted alkylene," "substituted alkenylene," "substituted alkynylene," "substituted heteroalkyl," "substituted heteroalkenyl," "substituted heteroalkynyl," "substituted aryl," "substituted heteroaryl," or "substituted heterocycloalkyl" refers to any group selected from the group consisting of C1-C4 10consisting of alkyl, halogen, perhaloalkyl, =O, -OH, alkoxy, tetrahydro-2-H-pyranyl, -NH2, -N(CH3)2, phenyl, benzyl, (1-methyl-imidazol-2-yl), pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, -C(=O)OH, trifluoromethyl, -C≡N, -C(=O)O(C1-C4)alkyl, -C(=O)NH2, -C(=O)NH(C1-C4)alkyl, -C(=O)N((C1-C4)alkyl)2, -SO2NH2, -C(=NH)NH2, and -NO2 By "alkyl", it is meant alkyl, cycloalkyl, alkenyl, alkynyl, alkylene, alkenylene, alkynylene, heteroalkyl, heteroalkenyl, heteroalkynyl, aryl, heteroaryl, or heterocycloalkyl as defined above, substituted with 1, 2, or 3 substituents selected from the group, preferably selected from halogen, -OH, alkoxy, -NH2, trifluoromethyl, -N(CH3)2, and -C(=O)OH, more preferably containing 1 or 2 substituents selected from halogen, alkoxy, and -OH. Examples of substituted alkyl include, but are not limited to, 2,2-difluoropropyl, 2-carboxycyclopentyl, and 3-chloropropyl.

[0049] As used herein, the term "alkoxy", employed alone or in combination with another term, unless otherwise specified, refers to an alkyl group as defined above having the specified number of carbon atoms and connected to the remainder of the molecule via an oxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy) and the higher homologs and isomers. (C1-C3)alkoxy, such as, but not limited to, ethoxy and methoxy, is preferred.

[0050] As used herein, the terms “halo” or “halogen,” by themselves or as part of another substituent, mean, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom, preferably fluorine, chlorine, or bromine, and more preferably fluorine or chlorine.

[0051] As used herein, the term "heteroalkyl" by itself or in combination with other terms means, unless otherwise specified, a stable straight or branched chain alkyl group consisting of the specified number of carbon atoms and one or two heteroatoms selected from the group consisting of O, N, and S, where the nitrogen and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. The heteroatom may be located at any position of the heteroalkyl group, including between the remainder of the heteroalkyl group and the fragment to which it is attached, or at the bond to the most distal carbon atom in the heteroalkyl group. Examples include -O-CH-CH-CH, -CH-CH-CH-OH, -CH-CH-NH-CH, -CH-S-CH-CH, and -CHCH-S(=O)-CH. Up to two heteroatoms may be consecutive, such as, for example, -CH-NH-OCH or -CH-CH-SS-CH.

[0052] As used herein, the term "heteroalkenyl" by itself or in combination with other terms refers to a stable straight or branched mono- or di-unsaturated hydrocarbon group consisting of the indicated number of carbon atoms and one or two heteroatoms selected from the group consisting of O, N, and S, unless otherwise specified, where the nitrogen and sulfur atoms are optionally oxidized and the nitrogen heteroatom is optionally quaternized. Up to two heteroatoms may be arranged consecutively. Examples include -CH=CH-O-CH3, -CH=CH-CH2-OH, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, and -CH2-CH=CH-CH2-SH.

[0053] As used herein, the term "aromatic" refers to a carbocyclic or heterocyclic ring with one or more polyunsaturated rings that have aromatic properties, i.e., have (4n+2) delocalized π (pi) electrons, where n is an integer.

[0054] As used herein, the term "aryl", employed alone or in combination with another term, unless otherwise specified, means a carbocyclic aromatic system containing one or more rings (typically 1, 2, or 3 rings), where such rings may be linked together in a pendant fashion, such as in biphenyl, or may be fused, such as in naphthalene. Examples include phenyl, anthracyl, and naphthyl. Phenyl and naphthyl are preferred, with phenyl being most preferred.

[0055] As used herein, the term "aryl-(C1-C3)alkyl" refers to a functional group in which an alkylene chain of 1-3 carbons is connected to an aryl group, e.g., -CH2CH2-phenyl or -CH2-phenyl(benzyl). Preferred are aryl-CH2- and aryl-CH(CH3)-. The term "substituted aryl-(C1-C3)alkyl" refers to an aryl-(C1-C3)alkyl functional group in which the aryl group is substituted. Preferred is substituted aryl(CH2)-. Similarly, the term "heteroaryl-(C1-C3)alkyl" refers to a functional group in which an alkylene chain of 1-3 carbons is connected to a heteroaryl group, e.g., -CH2CH2-pyridyl. Preferred is heteroaryl-(CH2)-. The term "substituted heteroaryl-(C1-C3)alkyl" refers to a heteroaryl-(C1-C3)alkyl functional group in which the heteroaryl group is substituted. Preferred is substituted heteroaryl-(CH2)-.

[0056] As used herein, the term "heterocycle" or "heterocyclyl" or "heterocyclic" by itself or as part of another substituent means, unless otherwise specified, a stable monocyclic or polycyclic heterocyclic ring system consisting of carbon atoms and at least one heteroatom selected from the group consisting of N, O, and S, which may be unsubstituted or substituted, where the nitrogen and sulfur heteroatoms may be optionally oxidized and the nitrogen atom may be optionally quaternized. The heterocyclic ring system may be attached to any heteroatom or carbon atom that results in a stable structure, unless otherwise specified. The heterocycle may be aromatic or non-aromatic in nature. In certain other embodiments, the heterocycle is a heteroaryl.

[0057] As used herein, the term "heteroaryl" or "heteroaromatic" refers to a heterocycle having aromatic character. Polycyclic heteroaryls may contain one or more rings that are partially saturated. Examples include tetrahydroquinoline and 2,3-dihydrobenzofuryl.

[0058] Examples of non-aromatic heterocycles include monocyclic groups such as aziridine, oxirane, thiirane, azetidine, oxetane, thietane, pyrrolidine, pyrroline, imidazoline, pyrazolidine, dioxolane, sulfolane, 2,3-dihydrofuran, 2,5-dihydrofuran, tetrahydrofuran, thiophane, piperidine, 1,2,3,6-tetrahydropyridine, 1,4-dihydropyridine, piperazine, morpholine, thiomorpholine, pyran, 2,3-dihydropyran, tetrahydropyran, 1,4-dioxane, 1,3-dioxane, homopiperazine, homopiperidine, 1,3-dioxepane, 4,7-dihydro-1,3-dioxepine, and hexamethylene oxide.

[0059] Examples of heteroaryl groups include pyridyl, pyrazinyl, pyrimidinyl (including, but not limited to, 2- and 4-pyrimidinyl), pyridazinyl, thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,3,4-thiadiazolyl and 1,3,4-oxadiazolyl.

[0060] Examples of polycyclic heterocycles are indolyl (including but not limited to 3-, 4-, 5-, 6-, and 7-indolyl), indolinyl, quinolyl, tetrahydroquinolyl, isoquinolyl (including but not limited to 1- and 5-isoquinolyl), 1,2,3,4-tetrahydroisoquinolyl, cinnolinyl, quinoxalinyl (including but not limited to 2- and 5-quinoxalinyl), quinazolinyl, phthalazinyl, 1,8-naphthyridinyl, 1,4-benzodioxanyl, coumarin, dihydrocoumarin, 1,5-naphthyridinyl, benzofuryl (including but not limited to ... including, but not limited to, 3-, 4-, 5-, 6-, and 7-benzofuryl), 2,3-dihydrobenzofuryl, 1,2-benzisoxazolyl, benzothienyl (including, but not limited to, 3-, 4-, 5-, 6-, and 7-benzothienyl), benzoxazolyl, benzothiazolyl (including, but not limited to, 2-benzothiazolyl and 5-benzothiazolyl), purinyl, benzimidazolyl, benzotriazolyl, thioxanthinyl, carbazolyl, carbolinyl, acridinyl, pyrrolidinyl, and quinolidinyl.

[0061] The heterocyclyl and heteroaryl moieties listed above are intended to be representative and non-limiting.

[0062] As used herein, the term "substituted" means that an atom or group of atoms replaces a hydrogen as a substituent attached to another group. Non-limiting examples of "substituted" groups include C1-C 10Includes alkyl, halogen, perhaloalkyl, =O, -OH, alkoxy, -NH2, -N(CH3)2, phenyl, benzyl, (1-methyl-imidazol-2-yl), pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, -C(=O)OH, -C≡N, -C(=O)O(C1-C4)alkyl, -C(=O)NH2, -C(=O)NH(C1-C4)alkyl, -C(=O)N((C1-C4)alkyl)2, -SO2NH2, -C(=NH)NH2, and -NO2.

[0063] For aryl, aryl-(C1-C3)alkyl, and heterocyclyl groups, the term "substituted" as applied to the rings of these groups refers to any level of substitution, i.e., mono-, di-, tri-, tetra-, or penta-substitution, where such substitution is permitted. The substituents are independently selected, and substitution may be at any chemically accessible position. In certain other embodiments, the number of substituents varies between 1 and 4. In other embodiments, the number of substituents varies between 1 and 3. In yet other embodiments, the number of substituents varies between 1 and 2. In yet other embodiments, the substituents are independently selected from C 1~6 Alkyl, -OH, C 1~6 The substituent is selected from the group consisting of alkoxy, halo, amino, acetamido and nitro. As used herein, when the substituent is an alkyl or alkoxy group, the carbon chain may be branched, straight or cyclic, with straight chain being preferred. As used herein, the terms "substituted heterocycle" and "substituted heteroaryl" refer to a heterocycle or heteroaryl group having one or more substituents, including halogen, CN, OH, NO2, amino, alkyl, cycloalkyl, carboxyalkyl (C(O)Oalkyl), trifluoroalkyl such as CF3, aryloxy, alkoxy, aryl, or heteroaryl. The substituted heterocycle or heteroaryl group may have 1, 2, 3, or 4 substituents.

[0064] Throughout this disclosure, various aspects of the invention may be expressed in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Thus, the description of a range should be construed as specifically disclosing all possible narrower ranges, individual numerical values ​​within that range, and, where appropriate, partial integers of numerical values ​​within the range. For example, the description of a range such as 1 to 6 should be considered to specifically disclose subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, and individual numbers within that range, e.g., 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0065] Compounds and Compositions The invention includes compounds of formula (I) or a salt, solvate, enantiomer, diastereoisomer, geometric isomer, or tautomer thereof: TIFF2024536861000003.tif23128 formula, X is CR 1 or N; Y is CR 2 or N; R 1 is H, optionally substituted C1-C6 alkyl (including optionally substituted benzyl), -C(=O)NR 6 R 6 , -C(=O)OR 6 , or R 8 and; R 2 is H or optionally substituted C1-C6 alkyl (including optionally substituted benzyl); R 3 is H, -CN, -C(=O)OR 6 , -C(=O)NR 6 R 6 , optionally substituted phenyl, or optionally substituted C1-C6 alkyl (including optionally substituted benzyl); R 4 -C(=O)OR 6 or R 8 and; R 5 is, at each occurrence, independently, optionally substituted C1-C6 alkyl (including optionally substituted benzyl), or optionally substituted phenyl; R 6 is independently, at each occurrence, H or optionally substituted C1-C6 alkyl (including optionally substituted benzyl), or two R 6 forms an optionally substituted 4- to 7-membered heterocyclyl; R 8 is -C(=O)NH(optionally substituted acyl), -N(optionally substituted acyl)C(=O)R 7 , -NR 6 C(=O)R 7 or -NR 6 C(=O)NR 6 R 7 and; R 7 each occurrence independently represents optionally substituted C1-C6 alkyl (including optionally substituted benzyl), optionally substituted cycloalkyl, CH(optionally substituted heterocyclyl)(R 5 ), CH(R 5 )(R 5 ), or an optionally substituted 4- to 7-membered heterocyclyl; or R 6 and R 7 together with the N atom to which they are both attached form an optionally substituted 4- to 7-membered heterocyclyl; However, (I) is R 8 Contains one.

[0066] In certain embodiments, the compound of formula (I) is TIFF2024536861000004.tif26131.

[0067] In certain embodiments, R 4 is R 8 It is.

[0068] In certain embodiments, R8 -NR 6 C(=O)R 7 and R 6 is H and R 7 is CH(CH2CH3)Ph.

[0069] In certain embodiments, R 8 -NR 6 C(=O)R 7 and R 6 is H and R 7 is CH(CH2CH3)(4-F-Ph).

[0070] In certain embodiments, R 8 -NR 6 C(=O)NR 6 R 7 and R 6 is H, and NR 6 R 7 Equal 4-phenyl-piperazin-1-yl.

[0071] In certain embodiments, R 8 -NR 6 C(=O)NR 6 R 7 and R 6 is H and NR 6 R 7 is 4-(2-pyridyl)-piperazin-1-yl.

[0072] In certain embodiments, R 3 -C(=O)OR 6 and R 6 is CH3.

[0073] In certain embodiments, R 3 -C(=O)OR 6 and R 6 is CH2CH3.

[0074] In certain embodiments, R 3 is -C(=O)NR 6 R 6 and NR 6 R 6is NH(CH2-aryl), where aryl is selected from the group consisting of phenyl, 4-fluorophenyl, 4-chlorophenyl, and 4-trifluoromethylphenyl.

[0075] In certain embodiments, R 1 teeth TIFF2024536861000005.tif32151: During the ceremony, R a1 and R a2 each, if present, is independently selected from the group consisting of H and optionally substituted C1-C6 alkyl; R b1 , R b2 , R b3 , R b4 , and R b5 each, if present, is independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 haloalkyl, halogen, CN, and NO2; R b1 , R b2 , R b3 , R b4 , and R b5 Two vicinal substituents selected from the group consisting of, together with the atoms to which they are attached, optionally substituted C2-C 10 may form a heterocycloalkyl; R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 are each independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 haloalkyl, and halogen, if present; R c1 , Rc2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 Two vicinal substituents selected from the group consisting of, together with the atom to which they are attached, optionally substituted C6-C 10 may form an aryl; A 1 C2 to C 10 Heteroaryl and optionally substituted C2-C 10 heterocycloalkyl; G 1 is a bond and C(R c6 )(R c7 ) selected from the group consisting of; G 2 is a bond and C(R c8 )(R c9 ) selected from the group consisting of; G 3 is a bond and C(R c10 )(R c11 ) selected from the group consisting of; Z 1 is N and CR c9 and Z 2 is N and CR b5 is selected from the group consisting of:

[0076] In certain embodiments, R 4 teeth TIFF2024536861000006.tif32151: During the ceremony, R a1 and R a2 each, if present, is independently selected from the group consisting of H and optionally substituted C1-C6 alkyl; R b1 , R b2 , R b3 , R b4, and R b5 are each independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 haloalkyl, halogen, CN, and NO2, if present; R b1 , R b2 , R b3 , R b4 , and R b5 Two vicinal substituents selected from the group consisting of, together with the atoms to which they are attached, optionally substituted C2-C 10 may form a heterocycloalkyl; R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 are each independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 haloalkyl, and halogen, if present; R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 Two vicinal substituents selected from the group consisting of, together with the atom to which they are attached, optionally substituted C6-C 10 may form an aryl; A 1 C2 to C 10 Heteroaryl and optionally substituted C2-C 10 heterocycloalkyl; G 1 is a bond and C(Rc6 )(R c7 ) selected from the group consisting of; G 2 is a bond and C(R c8 )(R c9 ) selected from the group consisting of; G 3 is a bond and C(R c10 )(R c11 ) selected from the group consisting of; Z 1 is N and CR c9 and Z 2 is N and CR b5 is selected from the group consisting of:

[0077] In certain embodiments, R a1 is H and R a2 is ethyl. In certain embodiments, R a1 is ethyl, and R a2 is H.

[0078] In certain embodiments, R b1 is H. In certain embodiments, R b1 is methyl. In certain embodiments, R b1 is OMe. In certain embodiments, R b1 is F. In certain embodiments, R b1 is Cl. In certain embodiments, R b1 is CF3. In certain embodiments, R b1 is CN. In certain embodiments, R b1 is NO2.

[0079] In certain embodiments, R b2 is H. In certain embodiments, R b2 is methyl. In certain embodiments, R b2 is OMe. In certain embodiments, R b2 is F. In certain embodiments, R b2 is Cl. In certain embodiments, R b2 is CF3. In certain embodiments, R b2 is CN. In certain embodiments, R b2is NO2.

[0080] In certain embodiments, R b3 is H. In certain embodiments, R b3 is methyl. In certain embodiments, R b3 is OMe. In certain embodiments, R b3 is F. In certain embodiments, R b3 is Cl. In certain embodiments, R b3 is CF3. In certain embodiments, R b3 is CN. In certain embodiments, R b3 is NO2.

[0081] In certain embodiments, R b4 is H. In certain embodiments, R b4 is methyl. In certain embodiments, R b4 is OMe. In certain embodiments, R b4 is F. In certain embodiments, R b4 is Cl. In certain embodiments, R b4 is CF3. In certain embodiments, R b4 is CN. In certain embodiments, R b4 is NO2.

[0082] In certain embodiments, R b5 is H. In certain embodiments, R b5 is methyl. In certain embodiments, R b5 is OMe. In certain embodiments, R b5 is F. In certain embodiments, R b5 is Cl. In certain embodiments, R b5 is CF3. In certain embodiments, R b5 is CN. In certain embodiments, R b5 is NO2.

[0083] In certain embodiments, R b1 , R b2 , R b3 , R b4 , and R b5 Two vicinal substituents selected from the group consisting of: combined to form methylenedioxy.

[0084] In certain embodiments, R c1 is H. In certain embodiments, R c1 is Ph. In certain embodiments, R c2 is H. In certain embodiments, R c2 is Ph. In certain embodiments, R c3 is H. In certain embodiments, R c3 is Ph. In certain embodiments, R c4 is H. In certain embodiments, R c4 is Ph. In certain embodiments, R c5 is H. In certain embodiments, R c5 is Ph. In certain embodiments, R c6 is H. In certain embodiments, R c6 is Ph. In certain embodiments, R c7 is H. In certain embodiments, R c7 is Ph. In certain embodiments, R c8 is H. In certain embodiments, R c8 is Ph. In certain embodiments, R c9 is H. In certain embodiments, R c9 is Ph. In certain embodiments, R c10 is H. In certain embodiments, R c10 is Ph. In certain embodiments, R c11 is H. In certain embodiments, R c11 is Ph.

[0085] In certain embodiments, R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 Two vicinal substituents selected from the group consisting of: combined to form a fused phenyl.

[0086] In certain embodiments, G 1 , G 2 , and G3 In certain embodiments, none of G 1 , G 2 , and G 3 In certain embodiments, one of G 1 , G 2 , and G 3 In certain embodiments, two of G 1 , G 2 , and G 3 Each of is a bond.

[0087] In certain embodiments, A 1 teeth TIFF2024536861000007.tif12128. In certain embodiments, A 1 teeth The file is TIFF2024536861000008.tif11128.

[0088] In certain embodiments, R 2 is an optionally substituted C1-C6 alkyl, R 3 is C(=O)OR 6 and R 1 and R 4 One of TIFF2024536861000009.tif24128 and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF3, CN, and NO2 is selected from the group consisting of R b1 , R b2 , R b3 , R b4 , and R b5 Two vicinal substituents selected from the group consisting of: combined to form methylenedioxy.

[0089] In certain embodiments, R 3 is an optionally substituted C1-C6 alkyl, R 2 is C(=O)OR 6 and R1 and R 4 One of TIFF2024536861000010.tif24128 and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF3, CN, and NO2 is selected from the group consisting of R b1 , R b2 , R b3 , R b4 , and R b5 Two vicinal substituents selected from the group consisting of: combined to form methylenedioxy.

[0090] In certain embodiments, R 2 is an optionally substituted C1-C6 alkyl, R 3 is CN and R 1 and R 4 One of TIFF2024536861000011.tif24128 and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF3, CN, and NO2 is selected from the group consisting of R b1 , R b2 , R b3 , R b4 , and R b5 Two vicinal substituents selected from the group consisting of: combined to form methylenedioxy.

[0091] In certain embodiments, R 2 is C(=O)OR 6 and R 1 and R 4 One of TIFF2024536861000012.tif24128, R 3 and R 1 or R 4Of these, not more than one is a C1-C6 alkyl.

[0092] In certain embodiments, R 1 is H and R 2 is H and R 3 is -C(=O)NR 6 R 6 and R 6 At most one occurrence of will be H.

[0093] In certain embodiments, R 1 teeth TIFF2024536861000013.tif24128, R 2 is methyl, R 3 is H and R 4 is selected from the group consisting of C(=O)O(C1 alkyl), C(=O)O(optionally substituted C3 alkyl), C(=O)O(optionally substituted C4 alkyl), C(=O)O(optionally substituted C5 alkyl), and C(=O)O(optionally substituted C6 alkyl).

[0094] In certain embodiments, R 4 teeth TIFF2024536861000014.tif24128, R 3 is methyl, R 2 is H and R 1 is selected from the group consisting of C(=O)O(C1 alkyl), C(=O)O(optionally substituted C3 alkyl), C(=O)O(optionally substituted C4 alkyl), C(=O)O(optionally substituted C5 alkyl), and C(=O)O(optionally substituted C6 alkyl).

[0095] In certain embodiments, R 1 is C(=O)NH2, and R 4 teeth TIFF2024536861000015.tif24128, G 1 , G 2 , and G 3 Of these, at most one is a bond, and Rc2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 A pair of vicinal substituents selected from the group consisting of, together with the atom to which they are attached, is C6-C 10 Forms an aryl.

[0096] In certain embodiments, R 1 is C(=O)NH2, and R 4 teeth TIFF2024536861000016.tif24128, G 1 , G 2 , and G 3 Of these, at most one is a bond, and R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 A pair of vicinal substituents selected from the group consisting of, together with the atom to which they are attached, is C6-C 10 Form an aryl.

[0097] In certain embodiments, Y is N and X is CR 1 and R 1 and R 4 One of TIFF2024536861000017.tif24128 and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF3, CN, and NO2 is selected from the group consisting of R b1 , R b2 , R b3 , R b4, and R b5 Two vicinal substituents selected from the group consisting of: combined to form methylenedioxy.

[0098] In certain embodiments, X is N and Y is CR 2 and R 4 teeth TIFF2024536861000018.tif24128 and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF3, CN, and NO2 is selected from the group consisting of R b1 , R b2 , R b3 , R b4 , and R b5 Two vicinal substituents selected from the group consisting of: combined to form methylenedioxy.

[0099] In certain embodiments, X is N, Y is N, and R 4 teeth TIFF2024536861000019.tif31128, Z 1 is CR c9 It is.

[0100] In certain embodiments, X is N, Y is N, and R 4 teeth TIFF2024536861000020.tif31128, Z 2 is CR b5 and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of is selected from the group consisting of halogen, C1-C6 alkyl, NO2, and CN.

[0101] In certain embodiments, R 1 is H. In certain embodiments, R 1is Me. In certain embodiments, R 1 is C(=O)NH2. In certain embodiments, R 1 is C(=O)NMe2. In certain embodiments, R 1 is C(=O)NEt2. In certain embodiments, R 1 is C(=O)OEt. In certain embodiments, R 1 is C(=O)OMe. In certain embodiments, R 1 is C(=O)OEt. In certain embodiments, R 1 is C(=O)Ot-Bu. In certain embodiments, R 1 is C(=O)O(CH2)2NH2. In certain embodiments, R 1 is C(=O)NH(CH2)3NH2. In certain embodiments, R 1 teeth TIFF2024536861000021.tif17128. In certain embodiments, R 1 teeth TIFF2024536861000022.tif17128. In certain embodiments, R 1 teeth TIFF2024536861000023.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000024.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000025.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000026.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000027.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000028.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000029.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000030.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000031.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000032.tif25128. In certain embodiments, R 1 teeth TIFF2024536861000033.tif20128. In certain embodiments, R 1 teeth TIFF2024536861000034.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000035.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000036.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000037.tif20128. In certain embodiments, R 1 teeth TIFF2024536861000038.tif19128. In certain embodiments, R 1 teeth TIFF2024536861000039.tif30128. In certain embodiments, R 1 teeth TIFF2024536861000040.tif15128. In certain embodiments, R 1 teeth TIFF2024536861000041.tif17128. In certain embodiments, R 1 teeth TIFF2024536861000042.tif16128. In certain embodiments, R 1 teeth TIFF2024536861000043.tif16128. In certain embodiments, R 1 teeth TIFF2024536861000044.tif16128. In certain embodiments, R 1 teeth TIFF2024536861000045.tif16128. In certain embodiments, R 1 teeth TIFF2024536861000046.tif17128. In certain embodiments, R 1 teeth TIFF2024536861000047.tif26128. In certain embodiments, R 1 teeth TIFF2024536861000048.tif26128. In certain embodiments, R 1 teeth TIFF2024536861000049.tif41128. In certain embodiments, R 1 teeth TIFF2024536861000050.tif15128. In certain embodiments, R 1 teeth TIFF2024536861000051.tif15128. In certain embodiments, R 1 teeth TIFF2024536861000052.tif25128. In certain embodiments, R 1 teeth TIFF2024536861000053.tif26128. In certain embodiments, R 1 teeth TIFF2024536861000054.tif25128. In certain embodiments, R 1 teeth TIFF2024536861000055.tif25128. In certain embodiments, R 1 teeth The file is TIFF2024536861000056.tif25128.

[0102] In certain embodiments, R 4 is Me. In certain embodiments, R 4 is C(=O)NH2. In certain embodiments, R 4is C(=O)NMe2. In certain embodiments, R 4 is C(=O)NEt2. In certain embodiments, R 4 is C(=O)OEt. In certain embodiments, R 4 is C(=O)OMe. In certain embodiments, R 4 is C(=O)OEt. In certain embodiments, R 4 is C(=O)Ot-Bu. In certain embodiments, R 4 is C(=O)O(CH2)2NH2. In certain embodiments, R 4 is C(=O)NH(CH2)3NH2. In certain embodiments, R 4 teeth TIFF2024536861000057.tif17128. In certain embodiments, R 4 teeth TIFF2024536861000058.tif17128. In certain embodiments, R 4 teeth TIFF2024536861000059.tif19128. In certain embodiments, R 4 teeth TIFF2024536861000060.tif19128. In certain embodiments, R 4 teeth TIFF2024536861000061.tif19128. In certain embodiments, R 4 teeth TIFF2024536861000062.tif19128. In certain embodiments, R 4 teeth TIFF2024536861000063.tif19128. In certain embodiments, R 4 teeth TIFF2024536861000064.tif20128. In certain embodiments, R 4 teeth TIFF2024536861000065.tif19128. In certain embodiments, R 4 teeth TIFF2024536861000066.tif19128. In certain embodiments, R 4 teeth TIFF2024536861000067.tif20128. In certain embodiments, R 4 teeth TIFF2024536861000068.tif25128. In certain embodiments, R 4 teeth TIFF2024536861000069.tif20128. In certain embodiments, R 4 teeth TIFF2024536861000070.tif19128. In certain embodiments, R 4 teeth TIFF2024536861000071.tif20128. In certain embodiments, R 4 teeth TIFF2024536861000072.tif20128. In certain embodiments, R 4 teeth TIFF2024536861000073.tif20128. In certain embodiments, R 4 teeth TIFF2024536861000074.tif19128. In certain embodiments, R 4 teeth TIFF2024536861000075.tif30128. In certain embodiments, R 4 teeth TIFF2024536861000076.tif15128. In certain embodiments, R 4 teeth TIFF2024536861000077.tif17128. In certain embodiments, R 4 teeth TIFF2024536861000078.tif16128. In certain embodiments, R 4 teeth TIFF2024536861000079.tif16128. In certain embodiments, R 4 teeth TIFF2024536861000080.tif16128. In certain embodiments, R 4 teeth TIFF2024536861000081.tif16128. In certain embodiments, R 4 teeth TIFF2024536861000082.tif17128. In certain embodiments, R 4 teeth TIFF2024536861000083.tif26128. In certain embodiments, R 4 teeth TIFF2024536861000084.tif26128. In certain embodiments, R 4 teeth TIFF2024536861000085.tif41128. In certain embodiments, R 4 teeth TIFF2024536861000086.tif15128. In certain embodiments, R 4 teeth TIFF2024536861000087.tif15128. In certain embodiments, R 4 teeth TIFF2024536861000088.tif25128. In certain embodiments, R 4 teeth TIFF2024536861000089.tif26128. In certain embodiments, R 4 teeth TIFF2024536861000090.tif25128. In certain embodiments, R 4 teeth TIFF2024536861000091.tif25128. In certain embodiments, R 4 teeth The file is TIFF2024536861000092.tif25128.

[0103] In certain embodiments, R 2 is H. In certain embodiments, R 2 is Me. In certain embodiments, R 2 is Et.

[0104] In certain embodiments, R 3 is H. In certain embodiments, R3 is Me. In certain embodiments, R 3 is Et. In certain embodiments, R 3 is C(=O)OMe. In certain embodiments, R 3 is C(=O)OEt. In certain embodiments, R 3 is C(=O)NH2. In certain embodiments, R 3 is CN. In certain embodiments, R 3 is Ph. In certain embodiments, R 3 is 4-trifluoromethylphenyl. In certain embodiments, R 3 is 4-fluorophenyl. In certain embodiments, R 3 teeth TIFF2024536861000093.tif17128. In certain embodiments, R 3 teeth TIFF2024536861000094.tif19128. In certain embodiments, R 3 teeth TIFF2024536861000095.tif19128. In certain embodiments, R 3 teeth TIFF2024536861000096.tif19128. In certain embodiments, R 3 teeth TIFF2024536861000097.tif17128. In certain embodiments, R 3 teeth TIFF2024536861000098.tif22128. In certain embodiments, R 3 teeth The file is TIFF2024536861000099.tif19128.

[0105] In certain further embodiments, the compound is not selected from the group consisting of 1, 2, 4, 5, 8, 9, 15, 16, 17, 19, 24, 26, 29, 31, 40, 66, 72, 76, 77, 78, 83, 84, and 86.

[0106] In certain further embodiments, the compound is selected from the group consisting of 1, 2, 4, 5, 8, 9, 15, 16, 17, 19, 24, 26, 31, 40, 66, 72, 76, 77, 78, 83, 84, and 86, 99, and 111.

[0107] In certain other embodiments, the compound is 22, 29, 32, 33, 34, 35, 36, 39, 47, 50, 57, 68, 75, 82, 87, 89, 90, 95, 96, 97, 98, 99, 106, 109, 110, 111, 112, 114, 115, 116, 118, 119, 123, 124, 127, 130, 131, 132, 1 34, 135, 144, 153, 154, 155, 159, 160, 161, 162, 163, 165, 167, 168, 169, 170, 172, 173, 174, 175, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, and 191.

[0108] The compounds described herein can form salts with acids and / or bases, and such salts are included in the present invention.In certain other embodiments, the salts are pharmaceutically acceptable salts.The term "salt" includes the addition salts of free acids and / or bases that are useful within the scope of the invention's method.Pharmaceutically unacceptable salts may nevertheless have properties such as high crystallinity, and are useful in the implementation of the present invention, for example, in the process of synthesis, purification or formulation of compounds that are useful within the scope of the invention's method.

[0109] Suitable pharma- ceutically acceptable acid addition salts may be prepared from inorganic acids or from organic acids. Examples of inorganic acids include sulfate, hydrogen sulfate, hemisulfate, hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid, and phosphoric acid (including hydrogen phosphate and dihydrogen phosphate). Suitable organic acids may be selected from the aliphatic, alicyclic, aromatic, aromatic aliphatic, heterocyclic, carboxylic, and sulfonic classes of organic acids, examples of which include formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, anthranilic acid, 4-hydroxybenzoic acid, phenylacetic acid, mannose, phenylacetic acid ... Derivative acid, embonic acid (pamoic acid), methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, pantothenic acid, trifluoromethanesulfonic acid, 2-hydroxyethanesulfonic acid, p-toluenesulfonic acid, sulfanilic acid, cyclohexylaminosulfonic acid, stearic acid, alginic acid, beta-hydroxybutyric acid, salicylic acid, galactaric acid, galacturonic acid, glycerophosphonic acid, and saccharin (e.g., saccharinate, saccharate).

[0110] Suitable pharma- ceutically acceptable base addition salts of the compounds of the invention include, for example, metallic salts including alkali metal, alkaline earth metal and transition metal salts, such as calcium, magnesium, potassium, sodium and zinc salts, etc. Pharmaceutically acceptable base addition salts also include organic salts made from basic amines, such as ammonium, N,N'-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine, etc.

[0111] All of these salts may be prepared, for example, from the corresponding compound by reacting the appropriate acid or base with the compound. The salt may contain a fraction less than one molar equivalent, a molar equivalent, or more than one molar equivalent of an acid or base relative to any compound of the invention. In certain alternative embodiments, at least one compound of the invention is a component of a pharmaceutical composition that further comprises at least one pharma- ceutically acceptable carrier.

[0112] The compounds of the invention may have one or more stereocenters, and each stereocenter may independently exist in either the (R) or (S) configuration. In certain alternative embodiments, the compounds described herein exist in optically active or racemic forms. The compounds described herein include racemic, optically active, regioisomeric and stereoisomeric forms, or combinations thereof, that have the therapeutically useful properties described herein. Preparation of optically active forms is accomplished in any suitable manner, including, but not limited to, resolution of racemic forms by recrystallization techniques, synthesis from optically active starting materials, chiral synthesis, or chromatographic separation using chiral stationary phases. In certain alternative embodiments, a mixture of one or more isomers is utilized as the therapeutic compounds described herein. In alternative embodiments, the compounds described herein contain one or more chiral centers. These compounds are prepared by any means, including stereoselective synthesis, enantioselective synthesis, and / or separation of mixtures of enantiomers and / or diastereomers. Resolution of the compounds and their isomers can be accomplished by any means, including, but not limited to, chemical processes, enzymatic processes, fractional crystallization, distillation, and chromatography.

[0113] The methods and formulations described herein include the use of N-oxides (if appropriate), crystalline forms (also known as polymorphs), solvates, amorphous phases, and / or pharmaceutically acceptable salts of compounds having the structure of any of the compounds of the present invention, as well as metabolites and active metabolites of these compounds having the same type of activity.Solvates include water, ether (e.g., tetrahydrofuran, methyl tert-butyl ether) or alcohol (e.g., ethanol) solvates, acetates, and the like. In certain other embodiments, the compounds described herein exist in solvated form with pharmaceutically acceptable solvents such as water and ethanol. In other embodiments, the compounds described herein exist in unsolvated form.

[0114] In certain alternative embodiments, the compounds of the invention exist as tautomers. All tautomers are included within the scope of the compounds provided herein.

[0115] In certain other embodiments, the compounds described herein are prepared as prodrugs. "Prodrugs" are drugs that are converted to parent drugs in vivo. In certain other embodiments, when administered in vivo, prodrugs are chemically converted to biologically, pharmacologic or therapeutically active forms of the compound. In other embodiments, prodrugs are enzymatically metabolized by one or more steps or processes to biologically, pharmacologic or therapeutically active forms of the compound.

[0116] In certain other embodiments, for example, sites on the aromatic ring portion of the compounds of the invention are susceptible to various metabolic reactions. Incorporation of appropriate substituents into the aromatic ring structure can reduce, minimize, or eliminate this metabolic pathway. In certain other embodiments, suitable substituents for reducing or eliminating the susceptibility of the aromatic ring to metabolic reactions are, by way of example only, deuterium, halogen, or alkyl groups.

[0117] The compounds described herein also include isotopically labeled compounds in which one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from that usually found in nature. Examples of isotopes that are suitably included in the compounds described herein are: 2 H, 3 H, 11 C. 13 C. 14 C. 36 Cl, 18 F, 123 I, 125 I, 13 N, 15 N, 15 O. 17 O. 18 O. 32 P, and 35 In certain other embodiments, isotope-labeled compounds are useful for drug and / or substrate tissue distribution studies. In other embodiments, substitution with heavier isotopes such as deuterium provides greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In yet other embodiments, 11 C. 18 F, 15 O and 13 Substitution with positron emitting isotopes, such as N, is useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically labeled compounds are prepared by any suitable method or process using an appropriate isotopically labeled reagent in place of an otherwise non-labeled reagent.

[0118] In certain alternative embodiments, the compounds described herein are labeled by alternative means, including but not limited to the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.

[0119] The compounds described herein and other related compounds with different substituents are synthesized using the techniques and materials described herein and in the art.The basic methods for the preparation of the compounds described herein are modified by the use of appropriate reagents and conditions for the introduction of the various moieties found in the formulas provided herein.

[0120] Pharmaceutical Compositions In one aspect, the disclosure provides pharmaceutical compositions comprising at least one compound of the invention and at least one pharma- ceutically acceptable carrier and / or excipient.

[0121] In one aspect, the disclosure provides a pharmaceutical composition comprising at least one compound selected from the group consisting of compounds 1, 2, 4, 5, 8, 9, 15, 16, 17, 19, 24, 26, 29, 31, 40, 66, 72, 76, 77, 78, 83, 84, 86, 99, and 111, and at least one pharma- ceutically acceptable excipient.

[0122] In certain embodiments, the at least one compound is compound 111.

[0123] In certain embodiments, the at least one pharma- ceutically acceptable excipient is selected from the group consisting of water, polyethylene glycol (PEG) 400, PEG 300, propylene glycol (PG), benzyl alcohol, polysorbate 80, diethylene glycol monoethyl ether (DEGEE), isopropyl myristate, ethanol, diisopropyl adipate, and lactate C. 12~15 At least one selected from the group consisting of alkyl, thickening agents, hydroxypropyl cellulose, and PEG 4000.

[0124] In certain embodiments, the thickening agent comprises a concentrated dispersion of a copolymer of acrylamide and sodium acryloyldimethyltaurate in isohexadecane. In certain embodiments, the thickening agent is SEPINEO™ P600.

[0125] In certain embodiments, the pharmaceutical composition comprises at least one of the following: (a) Compound 111, constituting about 0.1% to about 10.0% (w / w) of the pharmaceutical composition; (b) water, constituting about 10% to about 15% (w / w) of the pharmaceutical composition; (c) PEG 400 constituting about 20% to about 40% (w / w) of the pharmaceutical composition; (d) PEG300 constituting about 35% to about 60% (w / w) of the pharmaceutical composition; (e) PG constituting about 5% to about 15% (w / w) of the pharmaceutical composition; (f) benzyl alcohol, constituting from about 0.1% to about 5% (w / w) of the pharmaceutical composition; (g) polysorbate 80, constituting about 1% to about 10% (w / w) of the pharmaceutical composition; (h) a DEGEE constituting about 1% to about 15% (w / w) of the pharmaceutical composition; (i) isopropyl myristate, constituting about 0.1% to about 5% (w / w) of the pharmaceutical composition; (j) ethanol constituting about 5% to about 15% (w / w) of the pharmaceutical composition; (k) diisopropyl adipate, constituting about 5% to about 15% (w / w) of the pharmaceutical composition; (l) Lactic acid C, constituting about 5% to about 15% (w / w) of the pharmaceutical composition 12~15 Alkyl; (m) a viscosity enhancing agent comprising about 1% to about 10% (w / w) of the pharmaceutical composition; (n) hydroxypropylcellulose, constituting about 0.1% to about 5% (w / w) of the pharmaceutical composition; and (o) PEG 4000, constituting about 5% to about 15% (w / w) of the pharmaceutical composition.

[0126] In certain embodiments, the pharmaceutical composition comprises, consists of, or consists essentially of 111 (about 1.0% w / w), PEG400 (about 24.3% w / w), PEG300 (about 40.0% w / w), PG (about 10.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), isopropyl myristate (about 2.0% w / w), and PEG4000 (about 10% w / w).

[0127] In certain embodiments, the pharmaceutical composition comprises, consists of, or consists essentially of 111 (about 1.0% w / w), PEG400 (about 23.6% w / w), PEG300 (about 40.0% w / w), PG (about 10.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), and PEG4000 (about 10.0% w / w).

[0128] In certain embodiments, the pharmaceutical composition comprises, consists of, or consists essentially of 111 (about 1.0% w / w), PEG400 (about 34.0% w / w), PEG300 (about 40.0% w / w), PG (about 10% w / w), benzyl alcohol (about 2.0% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), isopropyl myristate (about 2.0% w / w), and hydroxypropylcellulose (about 1.0% w / w).

[0129] In certain embodiments, the pharmaceutical composition comprises, consists of, or consists essentially of 111 (about 1.0% w / w), water (about 13.3% w / w), PEG300 (about 57.0% w / w), PG (about 10% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), isopropyl myristate (about 2.0% w / w), and a viscosity enhancing agent (about 4% w / w).

[0130] In certain embodiments, the pharmaceutical composition comprises 111 (about 1.0% w / w), PEG300 (about 48.0% w / w), PG (about 10% w / w), benzyl alcohol (about 1.5% w / w), DEGEE (about 10% w / w), ethanol (about 8.5% w / w), diisopropyl adipate (about 10% w / w), lactate C 12~15 The composition may comprise, consist of, or consist essentially of alkyl (about 10% w / w), and hydroxypropyl cellulose (about 1.0% w / w).

[0131] In certain embodiments, the at least one compound is compound 99.

[0132] In certain embodiments, the at least one pharma- ceutically acceptable excipient is selected from the group consisting of water, PEG400, PG, benzyl alcohol, polysorbate 80, DEGEE, isopropyl myristate, ethanol, diisopropyl adipate, and lactate C. 12~15 At least one selected from the group consisting of alkyl, dimethyl isosorbide, PEG40 hydrogenated castor oil (HCO), hydroxypropyl cellulose, and PEG4000.

[0133] In certain embodiments, the pharmaceutical composition comprises at least one of the following: (a) Compound 99, constituting about 0.1% to about 10.0% (w / w) of the pharmaceutical composition; (b) water, constituting about 1% to about 10% (w / w) of the pharmaceutical composition; (c) PEG 400 constituting about 25% to about 35% (w / w) of the pharmaceutical composition; (d) PG constituting about 10% to about 30% (w / w) of the pharmaceutical composition; (e) benzyl alcohol, constituting about 0.1% to about 5% (w / w) of the pharmaceutical composition; (f) polysorbate 80, constituting about 1% to about 10% (w / w) of the pharmaceutical composition; (g) a DEGEE constituting about 10% to about 50% (w / w) of the pharmaceutical composition; (h) isopropyl myristate, constituting about 0.1% to about 5% (w / w) of the pharmaceutical composition; (i) ethanol constituting about 20% to about 35% (w / w) of the pharmaceutical composition; (j) diisopropyl adipate, constituting about 5% to about 15% (w / w) of the pharmaceutical composition; (k) lactic acid C, constituting about 1% to about 15% (w / w) of the pharmaceutical composition 12~15 Alkyl; (l) dimethylisosorbide, constituting about 5% to about 15% (w / w) of the pharmaceutical composition; (m) PEG40 HCO, constituting about 1% to about 10% (w / w) of the pharmaceutical composition; (n) hydroxypropylcellulose, constituting about 0.1% to about 5% (w / w) of the pharmaceutical composition; and (o) PEG 4000, constituting about 5% to about 15% (w / w) of the pharmaceutical composition.

[0134] In certain embodiments, the pharmaceutical composition comprises, consists of, or consists essentially of PEG 99 (about 1.0% w / w), PEG400 (about 28.3% w / w), PG (about 20.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 25.0% w / w), isopropyl myristate (about 2.0% w / w), dimethyl isosorbide (about 10.0% w / w), PEG40 HCO (about 5.0% w / w), and hydroxypropyl cellulose (about 1.0% w / w).

[0135] In certain embodiments, the pharmaceutical composition comprises, consists of, or consists essentially of PEG 99 (about 1.0% w / w), PEG400 (about 30.30% w / w), PG (about 20.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate (about 5.0% w / w), DEGEE (about 25.0% w / w), dimethyl isosorbide (about 10.0% w / w), PEG40 HCO (about 5.0% w / w), and hydroxypropyl cellulose (about 1.0% w / w).

[0136] In certain embodiments, the pharmaceutical composition comprises, consists of, or consists essentially of PEG 99 (about 1.0% w / w), PEG400 (about 19.30% w / w), PG (about 20.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate (about 5.0% w / w), DEGEE (about 25.0% w / w), isopropyl myristate (about 2.0% w / w), dimethyl isosorbide (about 10.0% w / w), PEG40 HCO (about 5.0% w / w), and PEG4000 (about 10.0% w / w).

[0137] In certain embodiments, the pharmaceutical composition comprises 99 (about 1.0% w / w), DEGEE (about 40.0% w / w), ethanol (about 28.0% w / w), diisopropyl adipate (about 10.0% w / w), C 12~15 It comprises, consists of, or consists essentially of alkyl lactate (about 10.0% w / w), dimethyl isosorbide (about 10.0% w / w), and hydroxypropyl cellulose (about 1.0% w / w).

[0138] In certain embodiments, the pharmaceutical composition comprises 99 (about 1.0% w / w), water (about 5.0% w / w), DEGEE (about 42.5% w / w), ethanol (about 25.0% w / w), diisopropyl adipate (about 10.0% w / w), C 12~15 It comprises, consists of, or consists essentially of alkyl lactate (about 5.0% w / w), dimethyl isosorbide (about 10.0% w / w), and hydroxypropyl cellulose (about 1.5% w / w).

[0139] In certain embodiments, the pharmaceutical composition is formulated for topical administration, hi certain embodiments, the topical formulation comprises a gel or ointment.

[0140] method The invention includes a method of treating, ameliorating, and / or preventing an orthopoxvirus infection in a human subject. In certain embodiments, the orthopoxvirus infection is caused by Molluscum contagiosum virus (MCV). In certain embodiments, the orthopoxvirus infection is caused by camelpox virus. In certain embodiments, the orthopoxvirus infection is caused by cowpox virus. In certain embodiments, the orthopoxvirus infection is caused by mousepox virus. In certain embodiments, the orthopoxvirus infection is caused by horsepox virus. In certain embodiments, the orthopoxvirus infection is caused by monkeypox virus. In certain embodiments, the orthopoxvirus infection is caused by raccoonpox virus. In certain embodiments, the orthopoxvirus infection is caused by tanapox virus. In certain embodiments, the orthopoxvirus infection is caused by smallpox (smallpox virus). In certain embodiments, the orthopoxvirus infection is caused by yokapox virus. In certain embodiments, the orthopoxvirus infection is caused by a cervidopoxvirus (deer pox). In certain embodiments, the orthopoxvirus infection is caused by an avipoxvirus (fowl pox). In certain embodiments, the orthopoxvirus infection is caused by a capripoxvirus (goat pox). In certain embodiments, the orthopoxvirus infection is caused by a leporipoxvirus (myxoma virus). In certain embodiments, the orthopoxvirus infection is caused by a parapoxvirus (orph virus). In certain embodiments, the orthopoxvirus infection is caused by a suipoxvirus (swinepox). In certain embodiments, the orthopoxvirus infection is caused by a batapoxvirus (yaba-like disease virus). In certain embodiments, the method comprises administering to the subject a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, solvate, enantiomer, diastereoisomer, geometric isomer, or tautomer thereof.

[0141] In certain embodiments, the compound of the present invention is a compound of formula (II): TIFF2024536861000100.tif23128X is CR 1 or N; Y is CR 2 or N; R 1 is H, optionally substituted C1-C6 alkyl, -C(=O)NR 6 R 6 , -C(=O)OR 6 , or R 8 and; R 2 is H or optionally substituted C1-C6 alkyl; R 3 H, -CN, -C(=O)OR 6 , -C(=O)NR 6 R 6 , optionally substituted phenyl, or optionally substituted C1-C6 alkyl; R 4 -C(=O)OR 6 or R 8 and; R 5 is optionally substituted C1-C6 alkyl or optionally substituted phenyl; R 6 is independently at each occurrence H or optionally substituted C1-C6 alkyl, or two R 6 are bonded to the same N to form an optionally substituted 4- to 7-membered heterocyclyl; R 8 is -C(=O)NH(optionally substituted acyl), -N(optionally substituted acyl)C(=O)R 7 , -NR 6 C(=O)R 7 or -NR 6 C(=O)NR 6 R 7 and R 7 is optionally substituted C1-C6 alkyl, optionally substituted cycloalkyl, CH(optionally substituted heterocyclyl)(R 5 ), CH(R5 )(R 5 ), or an optionally substituted 4- to 7-membered heterocyclyl, or R 6 and R 7 are bonded to the same N to form an optionally substituted 4- to 7-membered heterocyclyl; However, (I) is R 8 Contains one.

[0142] In certain embodiments, the compound or any composition comprising it is applied to the skin of an infected human. In another embodiment, the compound or any composition comprising it is applied to at least one MCV lesion on the skin of an infected human. In yet another embodiment, the compound is formulated as a topical pharmaceutical composition. In certain embodiments, the topical pharmaceutical composition comprises a gel or ointment. In yet another embodiment, the compound or any composition comprising it is administered topically to an infected human.

[0143] Administration / Dosage / Formulation The dosing regimen may affect the effective amount. The therapeutic formulation may be administered to the subject either before or after the onset of the disease or disorder contemplated in the invention. Furthermore, several divided doses or staggered doses may be administered daily or sequentially, or the dose may be continuously infused or bolus injected. Furthermore, the dosage of the therapeutic formulation may be increased or decreased in proportion to the exigencies of the therapeutic or prophylactic situation.

[0144] Administration of the compositions of the invention to a patient, preferably a mammal, more preferably a human, may be carried out using known procedures at dosages and for periods of time effective to treat the disease or disorder contemplated in the invention. The effective amount of the therapeutic compound required to achieve a therapeutic effect may vary depending on factors such as the state of the disease or disorder in the patient; the age, sex, and weight of the patient; and the ability of the therapeutic compound to treat the disease or disorder contemplated in the invention. Dosage regimens may be adjusted to provide an optimal therapeutic response. For example, several divided doses may be administered daily, or the dose may be reduced in proportion to the exigencies of the therapeutic situation. Non-limiting examples of effective dosage ranges for the therapeutic compounds of the invention are about 1-5,000 mg / kg body weight / day. Pharmaceutical compositions useful for carrying out the invention may be administered to deliver a dose of 1 ng / kg / day to 100 mg / kg / day. One of ordinary skill in the art would be able to consider the relevant factors and make the determination regarding the effective amount of the therapeutic compound without undue experimentation.

[0145] A medical practitioner, e.g., a physician or veterinarian, having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the invention employed in the pharmaceutical composition at levels lower than those required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0146] In a specific embodiment, it is advantageous to formulate the compound in dosage unit form for ease of administration and uniformity of dosage.As used herein, dosage unit form refers to a physically discrete unit suitable as a single dosage for the patient to be treated; each unit contains a predetermined amount of therapeutic compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle.

[0147] In certain other embodiments, the compositions of the invention are formulated with one or more pharma- ceutically acceptable excipients or carriers. In other embodiments, the pharmaceutical compositions of the invention comprise a therapeutically effective amount of the compounds of the invention and a pharma- ceutically acceptable carrier. In yet other embodiments, the compounds of the invention are the only biologically active agents in the composition (i.e., capable of treating, ameliorating, and / or preventing the diseases and disorders discussed herein). In yet other embodiments, the compounds of the invention are the only biologically active agents in a therapeutically effective amount in the composition (i.e., capable of treating, ameliorating, and / or preventing the diseases and disorders discussed herein).

[0148] In certain other embodiments, the compositions of the invention are administered to a patient in dosages ranging from 1 to 5 or more times per day. In other embodiments, the compositions of the invention are administered to a patient in dosages ranging from, but not limited to, once every day, every 2 or 3 days to once a week, and once every 2 weeks. It will be readily apparent to one of skill in the art that the frequency of administration of the various combination compositions of the invention will vary from individual to individual depending on many factors, including but not limited to, age, disease or disorder being treated, sex, general health, and other factors. Thus, the invention should not be construed as being limited to any particular dosing regimen, and the exact dosage and composition administered to any patient will be determined by the attending physician considering all other factors related to the patient.

[0149] The compounds of the invention for administration are from about 1 μg to about 10,000 mg, from about 20 μg to about 9,500 mg, from about 40 μg to about 9,000 mg, from about 75 μg to about 8,500 mg, from about 150 μg to about 7,500 mg, from about 200 μg to about 7,000 mg, from about 300 μg to about 6,000 mg, from about 500 μg to about 5,000 mg, from about 750 μg to about 4,000 mg, from about 1 mg to about 3,000 mg, from about 10 The range may be from about 2,500 mg, from about 20 mg to about 2,000 mg, from about 25 mg to about 1,500 mg, from about 30 mg to about 1,000 mg, from about 40 mg to about 900 mg, from about 50 mg to about 800 mg, from about 60 mg to about 750 mg, from about 70 mg to about 600 mg, from about 80 mg to about 500 mg, and any and all whole or partial increments therebetween.

[0150] In some embodiments, the dosage of the compound of the invention is from about 1 mg to about 2,500 mg. In some embodiments, the dosage of the compound of the invention used in the compositions described herein is less than about 10,000 mg, or less than about 8,000 mg, or less than about 6,000 mg, or less than about 5,000 mg, or less than about 3,000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, the dosage of a second compound described herein is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, and any and all full or partial increments thereof.

[0151] In certain other aspects, the invention is directed to a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of a compound of the invention, alone or in combination with a second pharmaceutical agent, and instructions for using the compound to treat, prevent, or reduce one or more symptoms of a disease or disorder contemplated in the invention.

[0152] The formulations may be employed in admixture with conventional excipients, i.e., pharma- ceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, intravenous, subcutaneous, enteral, or any other suitable mode of administration known in the art. Pharmaceutical preparations are sterilized and may be admixed, if desired, with auxiliary substances, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salt buffers for influencing osmotic pressure, coloring, flavoring and / or aromatic substances, etc. They may also be combined, if desired, with other active agents.

[0153] Routes of administration of any of the compositions of the invention include intravitreal, oral, nasal, rectal, intravaginal, parenteral, buccal, sublingual or topical routes. Compositions for use in the invention are formulated for administration by any suitable route, such as oral or parenteral, e.g., transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, ​​vaginal (e.g., vaginal and perivaginal), (intral)nasal and (trans)rectal), intravitreal, intravesical, intrapulmonary, intraduodenal, intragastric, intrathecal, subcutaneous, intramuscular, intradermal, intraarterial, intravenous, intrabronchial, inhalation, and topical administration.

[0154] Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gelcaps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosol formulations for inhalation, compositions and formulations for intravesical administration, etc. It should be understood that the formulations and compositions contemplated as useful in the present invention are not limited to the specific formulations and compositions described herein.

[0155] As used herein, "parenteral administration" of a pharmaceutical composition includes any route of administration characterized by physical breaching of a subject's tissue and administration of the pharmaceutical composition through the breach into the tissue. Thus, parenteral administration includes, but is not limited to, administration of a pharmaceutical composition, such as by injection of the composition, application of the composition through a surgical incision, or application of the composition through a non-surgical wound that penetrates the tissue. In particular, parenteral administration is intended to include, but is not limited to, subcutaneous, intravenous, intravitreal, intraperitoneal, intramuscular, intrasternal injection, and kidney dialysis infusion techniques.

[0156] Topical administration The obstacle to topical administration of pharmaceuticals is the stratum corneum of the epidermis. The stratum corneum is a highly resistant layer composed of proteins, cholesterol, sphingolipids, free fatty acids and various other lipids, and contains keratinocytes and living cells. One of the factors limiting the penetration rate (flux) of a compound through the stratum corneum is the amount of active substance that can be spread or applied onto the skin surface. The more active substance that is applied per unit area of ​​skin, the greater the concentration gradient between the skin surface and the lower layers of the skin, which in turn increases the diffusion force of the active substance through the skin. Thus, a formulation containing a higher concentration of active substance will have a higher chance of penetrating the active substance through the skin than a formulation with a lower concentration, all other things being equal, and the more, the more consistent the rate.

[0157] Formulations suitable for topical administration include, but are not limited to, liquid or semi-liquid preparations such as liniments, lotions, oil-in-water or water-in-oil emulsions, e.g., creams, ointments or pastes, and solutions or suspensions. Topically administrable formulations may contain, for example, about 1% to about 10% (w / w) active ingredient, although the concentration of the active ingredient may be as high as the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further include one or more of the additional ingredients described herein.

[0158] Penetration enhancers may be used. These substances increase the rate of penetration of drugs through the skin. Typical enhancers in the art include ethanol, glycerol monolaurate, PGML (polyethylene glycol monolaurate), dimethylsulfoxide, etc. Other enhancers include oleic acid, oleyl alcohol, ethoxydiglycol, laurocapram, alkanecarboxylic acids, dimethylsulfoxide, polar lipids, or N-methyl-2-pyrrolidone.

[0159] One acceptable vehicle for topical delivery of some of the compositions of the invention may contain liposomes. The composition of liposomes and their use are known in the art (see, e.g., U.S. Patent No. 6,323,219).

[0160] In alternative embodiments, the topically active pharmaceutical composition may be optionally combined with other ingredients, such as adjuvants, antioxidants, chelating agents, surfactants, foaming agents, wetting agents, emulsifiers, thickening agents, buffers, preservatives, etc. In another embodiment, a permeation or penetration enhancer is included in the composition, which is effective in enhancing the permeation of the active ingredient into and through the stratum corneum, compared to a composition that does not include the permeation enhancer. Various permeation enhancers are known to those skilled in the art, including oleic acid, oleyl alcohol, ethoxydiglycol, laurocapram, alkane carboxylic acid, dimethyl sulfoxide, polar lipids, or N-methyl-2-pyrrolidone. In another aspect, the composition may further include a hydrotropic agent, which functions to increase the disorder of the stratum corneum structure, thus allowing increased transport through the stratum corneum. Various hydrotropic agents are known to those skilled in the art, such as isopropyl alcohol, propylene glycol, or sodium xylene sulfonate.

[0161] Topically active pharmaceutical compositions should be applied in an effective amount to produce the desired change. As used herein, "effective amount" means an amount sufficient to cover the area of ​​the skin surface where the change is desired. The active compound should be present in an amount of about 0.0001% to about 15% by weight volume of the composition. More preferably, it should be present in an amount of about 0.0005% to about 5% by weight volume of the composition; most preferably, it should be present in an amount of about 0.001% to about 1% by weight volume of the composition. Such compounds may be synthetic or naturally derived.

[0162] Buccal administration The pharmaceutical compositions of the invention may be prepared, packaged, or sold in a formulation suitable for buccal administration. Such formulations may be, for example, in the form of tablets or lozenges made using conventional methods and may contain, for example, 0.1 to 20% (w / w) of the active ingredient, the remainder being an orally soluble or disintegrable composition, and optionally one or more additional ingredients described herein. Alternatively, a formulation suitable for buccal administration may comprise a powder or an aerosolized or atomized solution or suspension comprising the active ingredient. Such powdered, aerosolized, or atomized formulations, when dispersed, preferably have an average particle or droplet size in the range of about 0.1 to about 200 nanometers, and may further comprise one or more additional ingredients described herein. The exemplary formulations described herein are not intended to be exhaustive, and it is understood that the invention includes further modifications of these formulations, as well as other formulations not described herein but known to those of skill in the art.

[0163] Controlled Release Formulations and Drug Delivery Systems In certain alternative embodiments, the formulations of the present invention may be, but are not limited to, short-term release formulations, rapid-offset formulations, or controlled release formulations, including sustained release formulations, delayed release formulations, and pulsatile release formulations.

[0164] The term sustained release is used in its conventional sense to refer to a drug formulation that provides gradual release of drug over an extended period of time, resulting in, but not necessarily, substantially constant blood levels of drug over an extended period of time. The period may be as long as one month or more, and should be longer than the same amount of drug administered in bolus form. In certain embodiments, the compounds of the invention can be formulated for sustained release over a period of 3 to 12 months.

[0165] For sustained release, the compounds may be formulated with a suitable polymeric or hydrophobic material that provides sustained release properties to the compounds. Thus, the compounds useful within the methods of the invention may be administered in the form of microparticles, for example, by injection, or by implantation in the form of a wafer or disk.

[0166] In one embodiment of the invention, the compounds of the invention are administered to a patient, alone or in combination with other pharmaceutical agents, using sustained release formulations.

[0167] The term delayed release is used herein in its conventional sense to refer to drug formulations that provide an initial release of drug after some delay following drug administration, which may, but does not necessarily, include a delay of from about 10 minutes up to about 12 hours.

[0168] The term pulsatile release is used herein in its conventional sense to refer to a drug formulation that provides release of drug in such a manner as to produce a pulsatile plasma profile of drug following drug administration.

[0169] The term immediate release is used in its conventional sense to refer to a drug formulation that provides release of the drug immediately after drug administration.

[0170] As used herein, short-term refers to any period of time following drug administration of about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40 minutes, about 20 minutes, about 10 minutes, or about 1 minute or less, and any or all full or partial increments thereof.

[0171] As used herein, rapid off-action refers to any period of time following drug administration of about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40 minutes, about 20 minutes, about 10 minutes or less, or about 1 minute or less, and any and all total or partial increments thereof.

[0172] Dosage The therapeutically effective amount or dose of the compound of the present invention will vary depending on the age, sex and weight of the patient, the current medical condition of the patient, and the progression of the disease or disorder contemplated in the invention. Those skilled in the art will be able to determine the appropriate dosage depending on these and other factors.

[0173] A suitable dose of the compound of the present invention may range from about 0.01 mg to about 5,000 mg per day, for example, from about 0.1 mg to about 1,000 mg per day, for example, from about 1 mg to about 500 mg, for example, from about 5 mg to about 250 mg. The dose may be administered in a single dose or in multiple doses, for example, 1 to 5 or more times per day. When multiple doses are used, each dose may be the same or different. For example, a dose of 1 mg per day may be administered as two 0.5 mg doses, with an interval of about 12 hours between doses.

[0174] It is understood that the amount of compound administered per day can be administered, by way of non-limiting example, every day, every other day, every second day, every third day, every fourth day, or every fifth day.

[0175] If the patient's condition improves, administration of the inhibitor of the invention is optionally continued at the physician's discretion; alternatively, the dose of the administered drug is temporarily reduced or temporarily stopped (i.e., a "drug holiday") for a period of time. The length of the drug holiday can vary anywhere from 2 days to 1 year, including, by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. Dose reductions during drug holidays include 10% to 100%, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.

[0176] Once improvement of the patient's condition occurs, a maintenance dose is administered as needed.The dosage or frequency of administration, or both, is then reduced as a function of the disease or disorder to a level at which improvement of the disease is maintained.In certain other embodiments, the patient will require intermittent treatment on a long-term basis upon any recurrence of symptoms and / or infection.

[0177] The compounds for use in the methods of the invention may be formulated as unit dosage forms. The term "unit dosage form" refers to physically discrete units suitable as single dosages per patient undergoing treatment, each unit containing a predetermined amount of active material calculated to produce a desired therapeutic effect, optionally with a suitable pharmaceutical carrier. The unit dosage form may be for a single daily dose, or multiple daily doses (e.g., about 1 to 5 or more times per day). When multiple daily doses are used, the unit dosage form may be the same or different for each dose.

[0178] The toxicity and therapeutic efficacy of such treatment regimens are evaluated by LD 50 (a dose lethal to 50% of the population) and ED 50(the dose therapeutically effective in 50% of a population). The dose ratio between toxic and therapeutic effects is the therapeutic index, which is the LD 50 and ED 50 The data obtained from cell culture assays and animal studies are optionally used in formulating a range of dosages for use in humans. The dosage of such compounds is chosen so that they achieve the desired effect with minimal toxicity and the ED 50 The dosage may optionally vary within this range depending upon the dosage form employed and the route of administration utilized.

[0179] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents are considered to be within the scope of the present invention and encompassed by the claims appended hereto. For example, it should be understood that it is within the scope of this application to vary reaction conditions, including but not limited to reaction time, reaction scale / volume, and experimental reagents, such as solvents, catalysts, pressure, ambient conditions, such as nitrogen atmosphere, and reducing / oxidizing agents, with art-recognized alternatives and using no more than routine experimentation.

[0180] The following examples further illustrate aspects of the present invention, but do not in any way limit the teachings or disclosure of the invention set forth herein. EXAMPLES

[0181] The invention will now be described with reference to the following examples, which are provided for illustrative purposes only and the invention is not limited to these examples, but rather encompasses all variations that become evident as a result of the teachings provided herein.

[0182] material and method General Experimental Details Reactions were not performed under an inert atmosphere unless otherwise specified, and all solvents and commercially available reagents were used as received.

[0183] Cell handling BSC-1 and Vero cells were cultured in DMEM supplemented with 5% FBS, penicillin, and streptomycin (growth medium). Plaque reduction assays were performed in 48-well plates and cytotoxicity studies in 96-well plates.

[0184] compound All compounds were shown to be >95% pure by the supplier or by analysis, and unless otherwise noted, compounds were prepared as 10 mM solutions in DMSO.

[0185] LC / MS data (ESI+) were measured on a Waters Alliance 2695 HPLC / MS (Waters Symmetry C18, 4.6 × 75 mm, 3.5 μm) or (Phenomenex C18, 4.6 × 75 mm, 3.0 μm) equipped with a 2996 diode array detector at 210–400 nm; the solvent system was 5–95% MeCN in water (supplemented with 0.1% TFA) over 9 min using a linear gradient, with retention times in min. Mass spectrometry was performed on a Waters ZQ using electrospray in positive mode. Preparative reversed-phase HPLC was performed on a Waters Sunfire column (19 × 50 mm, C18, 5 μm) with detection wavelengths of 214 and 254 nm and a 10 min mobile phase gradient of 10% acetonitrile / water to 90% acetonitrile / water supplemented with 0.1% TFA as buffer. Injection and fraction collection were performed on a Gilson 215 liquid handler using Trilution LC software. 1 H NMR was recorded on a Varian Oxford 300 MHz. Chemical shifts (δ) are expressed in ppm downfield from tetramethylsilane (TMS) unless otherwise stated.

[0186] Plaque reduction assay, in vivo continuous DNA synthesis, and cytotoxicity Plaque reduction assays were performed on BSC-1 cells for VACV infection and on Vero cells for HSV-1 infection. Evaluation of continuous DNA synthesis was by ELISA-based rapid plate assay. Cytotoxicity was investigated by cell proliferation by seeding BSC-1 cells at approximately 10–15% confluence in 96-well plates and treating with 2-fold serially diluted compound solutions for 3 days. DMSO was kept at 1% throughout. Cell viability was measured by CyQUANT GR dye as recommended by the manufacturer (Invitrogen, USA). For 24-h treatment, BSC-1 cells were seeded at approximately 80% confluence and ATP production was monitored. Cells were removed from growth medium and lysed by adding 100 μL of 1% Triton X-100 PBS per well and incubating at room temperature for 10 min. Five microliters of lysate was used for luciferase / luminescence assays as recommended by the manufacturer (Invitrogen, USA).

[0187] Protein expression, purification, and handling Proteins were recombinantly expressed in Rosetta2pLysS and purified by Ni-NTA resin and gel filtration column chromatography. Unless otherwise noted, D4 proteins had their N-terminal His-tag removed by TEV protease and were maintained in elution column buffer: 20 mM sodium phosphate, pH 6.8, 0.2 M NaCl, and 15% w / v glycerol.

[0188] Differential scanning fluorescence analysis In a typical experiment, compounds were combined with 0.5 μM D4 in column buffer (reaction buffer) containing 2.5 mM DTT, 1% DMSO, and 0.005% Tween-20 in a reaction volume of 100 μL. After incubation at 25° C. for 20 min, 4 μL of 25× Sypro Orange (diluted in the same buffer to give a final concentration of approximately 0.96×) was added to the reaction mixture and centrifuged at 15,000 rpm for 1 min to remove particulates. 20 microliters of the supernatant was used for measurement and analysis.

[0189] Drug Affinity Responsive Target Stability (DARTS) DARTS experiments were adapted from Lomenick et al. (Lomenick, et al., 2009, PNAS USA 106(51):21984-21989). Bacterial cells expressing either N-terminal His (D4 and MBP-His8) or MBP tags were induced with 0.1 mM IPTG overnight at room temperature, and 3–5 mL of cell suspension was pelleted and used for the experiments. Cell pellets were resuspended in 500 μL of 20 mM sodium phosphate, pH 6.8, 0.2 M NaCl, and 0.5% Triton X-100 and lysed by sonication. After centrifugation at 15,000 rpm for 5 min, the cleared lysate was diluted 40-fold with reaction buffer to obtain an effective Triton X-100 concentration of approximately 0.012%. Typical reaction mixtures were performed in a volume of 100 μL. Proteolysis was performed with 5 μL of pronase (Calbiochem, USA; 50 mM Tris, pH 8, and 40 mM Ca 2+ This was achieved by the addition of 1:500 anti-His (GE Healthcare, USA) or 1:2000 anti-MBP (New England Biolabs, USA) antibodies.

[0190] Maleimide dye conjugation Cysteine ​​substitutions were introduced at positions 219 (Δ219C) and -19 (-19C; 19-aa upstream of the Met in D4 and immediately following the start codon) to allow conjugation to fluorescein-5-maleimide (AnaSpec Inc., USA) and N-(1-pyrene)maleimide (ThermoFisher Scientific, USA). The dyes were prepared as 10 mg / mL stock solutions in DMSO. During the gel filtration step of protein purification, the protein was treated with 10 mM DTT for 30 min at room temperature before loading onto the column. Protein eluates were kept below a 25 μM concentration and 4 molar equivalents of dye in column buffer containing 0.01% Triton X-100 and 5 mM EDTA were added immediately. The reactions were left overnight at 4°C protected from light. The protein / dye mixture was then centrifuged to remove particulates, and the supernatant was passed twice through Bio-Beads SM2 resin (Bio-Rad, USA) to remove Triton X-100. The eluate was further concentrated and purified by gel filtration to remove unbound dye and EDTA.

[0191] Isothermal titration calorimetry To generate the random DNA mix, fish sperm DNA (USB Scientific, USA) was dissolved in water and fragmented by sonication. The fragmented DNA was then dialyzed into column buffer without glycerol. 15mer oligomers containing poly d(TC) (Integrated DNA Technologies, USA) were prepared directly in the same buffer without the dialysis step. DNA and compounds were prepared in the same dialysis buffer with 2.5 mM DTT, 1% DMSO, and 0.005% Tween-20. DNA binding was then assessed by isothermal titration calorimetry (ITC) on a MicroCal iTC200 microcalorimeter (Malvern Instruments, United Kingdom) by titrating 3 μL per injection of 1.2 mg / mL random DNA (molarity unknown) or 5.3 mg / L single-stranded DNA into samples consisting of 40 μM compound. Experiments were performed at 25°C with 800 rpm agitation and 3 min intervals to equilibrate. Ethidium bromide (Sigma-Aldrich, USA) was freshly prepared in water. The molar concentrations of the DNA ligands were not defined, so only raw exotherms are shown.

[0192] Steady-state fluorescence Protein fluorescence was performed on a PTI photon counter equipped with a Model 810 detection system (HORIBA Scientific, USA). D4 was prepared at 0.5 μM in column buffer supplemented with 2.5 mM DTT and 0.005% Tween-20 and measured in a quartz cuvette. Tryptophan emission was monitored at 329 nm after excitation at 295 nm. To examine the effect of DMSO on tryptophan, DMSO was added last, mixed and emission was recorded for 30 min at 1 data point / s. All spectra were buffer subtracted to remove background fluorescence and scattering.

[0193] Surface plasmon resonance The experiments were carried out on a Biacore X100 (GE Healthcare Life Sciences, USA) at 25 °C using filtered and degassed buffer. Both flow cells of the NTA biosensor chip were filled with 0.5 mM Ni 2+ The cells were filled at 5 μL / min for 120 s and activated with a 1:1 mixture of 391 and 100 mM carbodiimide hydrochloride and N-hydroxysuccinimide, respectively, in 10 mM HEPES (pH 8), 200 mM NaCl, and 0.005% Tween-20 for 7 min. His-tagged MBP and D4 proteins were prepared at a concentration of 0.01 mg / mL in the same buffer containing 15% glycerol and injected for >30 min into reference and active cells, respectively, to achieve maximal cross-linking of proteins. After quenching with 0.5 M ethanolamine for 15 min, approximately 1000 and 3000 response units (RU) of MBP and D4, respectively, were obtained. For compound binding studies, the running buffer was 10 mM HEPES (pH 6.8), 200 mM NaCl, 0.5 mg / mL carboxymethyl dextran, 1% DMSO, 0.01% sodium azide, and 0.005% Tween-20. Compounds were dissolved in running buffer, sonicated for 10 min, and centrifuged at 15000 rpm for 5 min. A typical experiment consisted of injecting compound at 10 μL / min for 60 s and monitoring for 180 s, followed by an injection of 1% DMSO alone using the same procedure. Data were analyzed by the accompanying BIA evaluation software (GE Healthcare Life Sciences, USA). Each 1% DMSO injection was used for double referencing with the corresponding compound injection, and the equilibrium response (R eq ) values ​​were obtained by linear regression at steady state, whereby negative equilibrium values ​​were not used in the fitting. Experiments were repeated twice.

[0194] Continuous DNA synthesis assay Processive DNA synthesis was assessed using a rapid plate assay (Lin & Ricciardi, 2000, J. Virol. Methods 88:219-225). 5'-biotinylated 100 nucleotide templates containing an adenine only at their 5' distal end were annealed to a 15 nucleotide primer at their 3' end and attached to streptavidin-coated 96-plate wells. DNA synthesis was carried out in a 50 μL reaction mixture containing 100 mM (NH)2SO4, 20 mM Tris-HCl (pH 7.5), 3 mM MgCl2, 0.1 mM EDTA, 0.5 mM DTT, 2% glycerol, 40 μg / ml BSA, 5 μM dATP, 5 μM dCTP, 5 μM dGTP, 1 μM digoxigenin-11-dUTP, and E9 / A20 / D4 protein. TNT reticulocyte lysate or in vitro translated luciferase was used as negative controls. After 30 min incubation at 37°C, plates were washed extensively with phosphate-buffered saline (PBS). Wells were then incubated with anti-digoxigenin-peroxidase antibody for 1 h at 37°C and then washed with PBS. The substrate 2,2'-azino-bis(3-ethylbenzthiazoline)-sulfonate was added and color developed by gently rocking the plate. DNA synthesis was quantified by measuring the absorbance of each reaction at 405 nm with a microplate reader. Experiments were performed in triplicate and repeated at least twice independently.

[0195] Thermal shift assay Thermal shift (differential scanning fluorimetry) assays were performed as previously described (Nuth, et al., 2011, J. Med. Chem. 54:3260-3267). Briefly, 5 μM of purified 6His-mD4 was mixed with compounds in a thin-walled PCR96 plate well in a total volume of 20 μL containing 25 mM phosphate buffer (pH 6.8), 0.2 M NaCl, 2.5% glycerol, 2% DMSO, 0.005% (w / v) Triton-X100, and 1× Sypro Orange. Fluorescence intensity was monitored using an Applied Biosystems 7500 Fast Real-Time PCR system at a rate of 1°C / min, 25–80°C, and a wavelength of 582 nm. Melting temperature (T m Protein melting curves were plotted in GraphPad Prism and fitted to the Boltzmann sigmoidal model to generate thermal shifts (ΔT m ) is the difference between 2% DMSO mock treatment and inhibitor treatment. All experiments were performed twice and repeated independently.

[0196] Viral plaque reduction and cytotoxicity assays Viral plaque reduction assays were performed in triplicate using BSC-1 cells (Nuth, et al., 2011, J. Med. Chem. 54:3260-3267). Briefly, cells were infected by adsorption of virus at 80 PFU / well in 100 μL growth medium in 48-well plates for 1 h and then treated with compounds for 16 h. Cells were stained, plaques were counted under a dissecting microscope, and data were plotted in GraphPad Prism.

[0197] Franz Cell Apparatus The Franz cell apparatus and receptor medium are equilibrated to 35°C ± 1°C. Dermatome skin from a human cadaver torso is thawed and its integrity is checked by measuring the water loss from the epidermis with a water transpiration meter. After assembly of the apparatus, the skin and medium are allowed to equilibrate for 30 minutes. The skin temperature is checked with an infrared laser thermometer and a pre-dose (T = 0) sample is taken from the receptor medium. The mixed formulation (10 mg API / mL) is applied to the skin surface (1 cm 2 The area is then filled with 0.01 mL of medium) and the time of administration is recorded. Then, 0.5 mL samples of receptor medium are taken at 2, 4, 6, 21 and 24 hours after administration, and new warm medium is added to make up the volume of the sample. After the last time point, the surface of the skin is washed with 0.5 mL of PBS to remove residual formulation, and the surface is wiped dry with a cotton swab. The washing procedure is repeated twice. The skin is removed from the apparatus, spread on a flat surface, and the stratum corneum is removed by tape stripping (typically 4 strips). The tape strips are rinsed to recover the stratum corneum. The peeled skin is then placed on a heat block (preheated to 60 °C) for 1-2 minutes to heat separate the skin layers. The epidermis is peeled off using forceps, and the epidermal and dermal skin layers are weighed and homogenized in 1x PBS / 4% BSA. Protein-extracted samples are stored at -20 °C until further analysis by LC-MS / MS.

[0198] Example 1: Protein Dynamics Previous thermodynamic results suggested that D4 possesses protein flexibility that contributes to the global protein dynamics. This protein dynamics was speculated to be necessary for protein function and to be responsible for the observed heterogeneity in protein size in vitro. Compounds that can perturb the dynamics could therefore be used as design leads. For this reason, we needed to confirm that D4 indeed exhibits dynamic properties.

[0199] As a first approach, we investigated the effect of DMSO, the preferred organic solvent for compound preparation, on overall intrinsic tryptophan fluorescence. As tryptophan emission is sensitive to solvent exposure, such an approach can provide insight into the environment surrounding the five intrinsic tryptophan residues. Furthermore, it was shown that the addition of ethanol is prone to promoting protein aggregation, raising concerns that compound binding experiments with DMSO might be hindered by protein heterogeneity, so it was also important to ensure that DMSO did not adversely affect the protein structure.

[0200] Tryptophan emission was monitored over a period of 30 min, a typical period for incubation of proteins with compounds, in increasing DMSO concentrations. As shown in Figure 10A, a significant decrease in emission was observed between 0.5 and 5% DMSO. In contrast, there was no similar trend for the well-folded maltose binding protein (MBP). Given that tryptophan quenching is mediated by a solvent-stabilized charge transfer from the ring to the peptide backbone, this suggests that the observed fluorescence fluctuations likely arose from DMSO-H2O exchange at the protein surface, which could possibly be accelerated by a flexible protein. Examination of the fluorescence spectra of both D4 and MBP in 5% DMSO compared to 0% DMSO showed no evidence of a spectral shift, thus ruling out disruption of protein folding by DMSO (Figure 10A, inset).

[0201] In certain embodiments, proteins that exhibit flexibility / dynamics are more susceptible to digestion by proteases due to the transient exposure of buried sites. To test this hypothesis, bacterial lysates expressing a protein of interest were subjected to drug affinity responsive target stability ( d rug a ffinity r esponsive t target sThe proteins were exposed to various concentrations of the nonspecific protease pronase according to the Dynamics of Artificial Reaction Trouble (DARTS) setup and probed by Western blotting. Specifically, example proteins were selected based on their relative protein fold numbers. As shown in Figure 10B, D4 was barely detectable at the pronase dilution of 1:150 tested (corresponding to 3.2 μg / mL of protease in the reaction mixture). In comparison, well-folded MBP showed reasonable protection against proteolysis even at a pronase dilution of 1:37.5 (Figure 10B, corresponding to 12.7 μg / mL of protease). Addition of the N-terminal 63 amino acid portion of A20 to MBP subsequently rendered the protein more susceptible to proteolysis at the same pronase dilution (Figure 10B), whereas A20 63 This lends credence to the suggested disordered / unfolded state of D4. To compare the sensitivity of proteins with known flexibility / kinetics to pronase digestion, an MBP fusion protein of the N-terminal 103-aa of human estrogen receptor beta (ERβ-N) was constructed; in the absence of carrier protein, it was expressed as an inclusion body that could refold in vitro into an intrinsically disordered structure. As an MBP fusion, ERβ-N was detected as a minor product in crude cell lysates and was highly susceptible to protease digestion even at a pronase dilution of 1:1200 (Figure 10B). Taken together, the solvent exposure and DARTS results consistently demonstrated protein kinetics and indicated that D4 does not exhibit properties comparable to well-folded proteins such as MBP or those with obvious disordered structures.

[0202] Example 2: Compound 1 inhibits poxvirus DNA synthesis In vitro processive DNA synthesis was assessed by combining DNA polymerase and processivity factors in an ELISA-based assay. For testing VACV, DNA polymerase, A20, and D4 proteins were combined, while for HSV-1, UL30 (DNA polymerase) and UL42 (processivity factor) were combined. Consistent with the lack of antiviral activity against HSV-1 infection by compound 1 (Figure 8B), no inhibition of DNA synthesis was observed using HSV-1 proteins (Figure 8A). In contrast, compound 1 was able to inhibit DNA synthesis using poxvirus proteins. Specifically, when D4 of molluscum contagiosum virus (MCV) was substituted for the VACV reaction (i.e., DNA polymerase and A20 in the absence of VACV D4), inhibition of DNA synthesis was observed (IC 50 = 13.4 μM) (Figure 6). Nearly identical inhibition was observed for VACV.

[0203] Example 3: D4 is the target of compound 1 To investigate D4 as the intended target of compound 1, protein binding was first examined by incubating the compound with purified D4 protein and measured by differential scanning fluorometry (DSF). With increasing compound, a dose-dependent decrease in thermal shift was observed (ΔTm = -0.86 and -1.55 for 25 and 50 μM treatments, respectively), with all curves for compound treatments up to 50 μM showing comparable maximum fluorescence signals, indicating minimal compound-induced assay interference (e.g., protein precipitation) (Figure 12A, Table 1). For comparison, the drugs cidofovir (CDV) and tecovirimat (also known as ST-246), both of which have been shown to be effective against various poxviruses, did not show any thermal shift (within experimental error) up to 50 μM (Table 1).

[0204] Compound binding was then examined by surface plasmon resonance (SPR). Using the sensor chip NTA, his-tagged D4 was captured and crosslinked to a Ni-loaded active flow cell, while his-tagged MBP was similarly prepared for the reference flow cell and served as the corresponding irrelevant protein surface. As shown in Figure 12B, a dose-response of compound 1 was observed for binding to D4, with the binding affinity K estimated by steady-state analysis. D = 22.8 μM was obtained. In comparison, near-baseline signals and lack of dose response for compounds up to 50 μM were observed for both CDV and ST-246, with CDV showing negligible binding to the active flow cell at concentrations of 25 μM and 50 μM (Figure 12B). To confirm the specificity of compound 1 for D4, compound binding was further investigated by DARTS by incubating test compounds with crude cell lysates expressing D4 or control proteins. Given that the degree of protein folding determines the efficiency of proteolysis (Figure 10B), pronase was added in various amounts to obtain discernible differences (typically >50%) in proteolysis between untreated and mock-treated (Figures 12C-12D). As compound binding prevents proteolysis of the intended target, an increase in D4 protein levels was observed with increasing concentrations of compound 1 compared to mock-treated, consistent with D4 as a target (Figure 12C). In contrast, CDV and ST-246 had no effect on D4 protein levels at concentrations up to 100 μM (Figure S6D).Similarly, the inability of compound 1 to reduce levels of the unrelated proteins MBP and ERβ-N provided further evidence of specificity for D4 (Figure S6C).

[0205] Compound 1 was examined for promiscuous DNA binding, as this could have inadvertently contributed to the observed inhibition of in vitro DNA synthesis. The use of sheared DNA served as a source of random double-stranded DNA, and a 15mer poly d(TC) oligomer as a source of single-stranded DNA, as measured by isothermal titration calorimetry (ITC). Injection of either double-stranded or single-stranded DNA into samples containing compound 1 produced heat traces showing no appreciable total heat and no signal saturation, both attributes corresponding to the heat of dilution produced by injection of buffer alone (Figure 13), thus negating DNA binding by compound 1 to both DNA types. As a positive control, appreciable heat and saturation signals can be observed for the DNA intercalating ethidium bromide, which binds to double-stranded DNA (Figure 13). Taken together, the various approaches support the binding of compound 1 to D4, in addition to demonstrating the absence of promiscuous binding to unrelated proteins and DNA.

[0206] Example 4: Synthesis of selected compounds of the invention. Compound 22. 3-Methyl-5-(2-phenyl-butyrylamino)-thiophene-2-carboxylic acid methyl ester 2-Phenylbutyric acid (0.178 g, 1.08 mmol) was dissolved in anhydrous dichloromethane (2 mL). Oxalyl chloride (2.0 M in dichloromethane, 0.54 mL, 1.08 mmol) was added followed by one drop of dimethylformamide. The reaction was stirred at room temperature for 2 h and then evaporated to dryness. The residue was dissolved in anhydrous pyridine (2 mL). 5-Amino-3-methyl-thiophene-2-carboxylic acid methyl ester hydrochloride (0.075 g, 0.36 mmol) and 4-dimethylaminopyridine (0.004 g, 0.036 mmol) were added. The reaction was stirred at room temperature for 16 h. It was then diluted with brine and extracted with ethyl acetate. The extract was concentrated and subjected to chromatography (12 g column, 10-40% ethyl acetate in hexanes) to give 109 mg of product as a white solid. MS: [M+H] + 318.15.

[0207] Compound 29. Methyl 2-(2-phenylbutanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate 2-Phenylbutanoic acid (80 mg, 0.44 mmol) was taken up in 3 mL of dry DCM under N2 atmosphere. 1.2 equivalents of oxalyl chloride (45 μL, 53 mmol) was added to the solution along with one drop of dry DMF. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The acyl chloride was taken up in 3 mL of pyridine and 5-amino-3-methyl-4-propionylthiophene-2-carboxamide (25 mg, 0.13 mmol) was added. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, followed by washing with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by normal phase chromatography (0-10% MeOH / DCM). 9 mg (19% yield) of solid was produced. MS [M+H] + = 361.

[0208] Compound 32. 4-Methyl-2-(2-phenyl-butyrylamino)-thiophene-3-carboxylic acid methyl ester A mixture of 2-phenyl-butyric acid (266 mg, 1.62 mmol) and 2M oxalyl chloride in dichloromethane (0.77 mL, 1.54 mmol) was treated with DMF (3 drops). The reaction was stirred for 2 hours and then concentrated. The residue was treated with pyridine (0.5 mL), 2-amino-4-methyl-thiophene-3-carboxylic acid ethyl ester (100 mg, 0.54 mmol), and DMAP (5 mg). The mixture was stirred for 16 hours. The mixture was treated with ethyl acetate (30 mL), then washed with 1N HCl (2×20 mL), water (5 mL), and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by column on silica (0-10% ethyl acetate:hexanes). The nearly pure material was dissolved in methanol (2 mL) and treated with 6N NaOH (0.2 mL, 1.2 mmol). The reaction was stirred for 2 hours and then concentrated. The residue was treated with water (5 mL) and ethyl acetate (20 mL). The organic layer was washed with brine (1 mL), dried (Na2SO4) and concentrated. The crude material was purified by column on silica (0-10% ethyl acetate:hexanes) to give 4-methyl-2-(2-phenyl-butyrylamino)-thiophene-3-carboxylic acid methyl ester (28 mg, 16%) as a clear sticky gum. MS: [M+H] + 318.

[0209] Compound 33. 5-Carbamoyl-2-[2-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-3-phenyl-propionylamino]-4-methyl-thiophene-3-carboxylic acid methyl ester 2-(1,3-Dioxo-1,3-dihydro-isoindol-2-yl)-3-phenyl-propionic acid (0.413 g, 1.40 mmol) was dissolved in anhydrous dichloromethane (6 mL). Oxalyl chloride (2.0 M in dichloromethane, 0.70 mL, 1.40 mmol) was added followed by one drop of dimethylformamide. The reaction was stirred at room temperature for 1 hour and then evaporated to dryness. The residue was dissolved in anhydrous pyridine (5 mL). 2-Amino-5-carbamoyl-4-methyl-thiophene-3-carboxylic acid methyl ester (0.100 g, 0.467 mmol) and 4-dimethylaminopyridine (0.006 g, 0.047 mmol) were added. The reaction was stirred at room temperature for 16 hours. It was then diluted with saturated aqueous sodium bicarbonate and extracted with ethyl acetate. The extract was concentrated and chromatographed (12 g column, 10-40% ethyl acetate in hexanes) to give a white solid. The solid was dissolved in diethyl ether and hydrogen chloride (1.0 M in diethyl ether) was added. The resulting suspension was evaporated to dryness to give the hydrochloride salt as a white solid (58 mg). MS: [M+H] + 492.6.

[0210] Compound 34. N-(4-methyl-thiophen-2-yl)-2-phenyl-butyramide A mixture of 2-phenyl-butyric acid (73 mg, 0.44 mmol), EDC.HCl (126 mg, 0.66 mmol), and HOAt (90 mg, 0.66 mmol) in DMF (2 mL) was stirred for 5 min, then 4-methyl-thiophen-2-ylamine (50 mg, 0.44 mmol) was added. The reaction was stirred for 3 days. The mixture was treated with water (30 mL) and then extracted with ethyl acetate (2×20 mL). The combined organic extracts were washed with water (2×10 mL) and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by column on silica (0-30% ethyl acetate:hexanes) to give N-(4-methyl-thiophen-2-yl)-2-phenyl-butyramide (15 mg, 13%) as a grey solid. MS: [M+H] + 260.

[0211] Compound 35. 5-Carbamoyl-2-[(indan-1-carbonyl)-amino]-4-methyl-thiophene-3-carboxylic acid methyl ester A mixture of indan-1-carboxylic acid (131 mg, 0.81 mmol) in 2M oxalyl chloride in dichloromethane (0.39 mL, 0.78 mmol) was treated with DMF (2 drops). The reaction was stirred for 2 h and then concentrated. The residue was treated with pyridine (0.5 mL), 2-amino-5-carbamoyl-4-methyl-thiophene-3-carboxylic acid methyl ester (58 mg, 0.27 mmol), and DMAP (5 mg). The mixture was stirred for 16 h. The mixture was treated with ethyl acetate (30 mL) and then washed with 1N HCl (2×20 mL), water (5 mL), and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by column on silica (0-100% ethyl acetate:hexanes) to give 5-carbamoyl-2-[(indan-1-carbonyl)-amino]-4-methyl-thiophene-3-carboxylic acid methyl ester (47 mg, 49%) as a white solid. MS: [M+H] + 359.

[0212] Compound 36. 5-Carbamoyl-4-methyl-2-[(1-phenyl-cyclopropanecarbonyl)-amino]-thiophene-3-carboxylic acid methyl ester A mixture of 1-phenyl-cyclopropanecarboxylic acid (262 mg, 1.62 mmol) in 2M oxalyl chloride in dichloromethane (0.77 mL, 1.54 mmol) was treated with DMF (3 drops). The reaction was stirred for 2 h and then concentrated. The residue was treated with pyridine (0.5 mL), 2-amino-5-carbamoyl-4-methyl-thiophene-3-carboxylic acid methyl ester (116 mg, 0.54 mmol), and DMAP (5 mg). The mixture was stirred for 16 h. The mixture was treated with ethyl acetate (30 mL) and then washed with 1N HCl (2×20 mL), water (5 mL), and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by column on silica (0-100% ethyl acetate:hexanes) to give 5-carbamoyl-4-methyl-2-[(1-phenyl-cyclopropanecarbonyl)-amino]-thiophene-3-carboxylic acid methyl ester (13 mg, 7%) as an off-white solid. MS: [M+H] + 359.

[0213] Compound 39. N-(3-cyano-4,5-dimethyl-thiophen-2-yl)-2-phenyl-N-(2-phenyl-butyryl)-butyramide A mixture of 2-phenyl-butyric acid (266 mg, 1.62 mmol) in 2M oxalyl chloride in dichloromethane (0.77 mL, 1.54 mmol) was treated with DMF (3 drops). The reaction was stirred for 2 h and then concentrated. The residue was dissolved in dichloromethane (2.5 mL) and then treated with diisopropylethylamine (137 mg, 1.08 mmol) and 2-amino-4,5-dimethyl-thiophene-3-carbonitrile (82 mg, 0.54 mmol). The reaction was stirred for 16 h. The mixture was treated with water (30 mL) and then extracted with ethyl acetate (2×20 mL). The combined organic extracts were washed with water (2×10 mL) and brine (1 mL), dried (Na2SO4) and concentrated. The crude material was purified by column on silica (0-30% ethyl acetate:hexane) to give N-(3-cyano-4,5-dimethyl-thiophen-2-yl)-2-phenyl-N-(2-phenyl-butyryl)-butyramide (25 mg, 10%) as a grey solid. MS: [M+H] + 445.

[0214] Compound 47. 5-[bis-(1,2,3,4-tetrahydro-naphthalene-1-carbonyl)-amino]-4-cyano-3-methyl-thiophene-2-carboxylic acid amide A mixture of 1,2,3,4-tetrahydro-naphthalene-1-carboxylic acid (211 mg, 1.20 mmol) in 2M oxalyl chloride in dichloromethane (0.60 mL, 1.20 mmol) was treated with DMF (3 drops). The reaction was stirred for 2 h and then concentrated. The residue was treated with pyridine (0.5 mL), 5-amino-4-cyano-3-methyl-thiophene-2-carboxylic acid amide (72 mg, 0.40 mmol), and DMAP (5 mg). The mixture was stirred for 16 h. The mixture was treated with ethyl acetate (30 mL) and then washed with 1N HCl (2×20 mL), water (10 mL), and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by column on silica (0-100% ethyl acetate:hexanes) to give 5-[bis-(1,2,3,4-tetrahydro-naphthalene-1-carbonyl)-amino]-4-cyano-3-methyl-thiophene-2-carboxylic acid amide (11 mg, 6%) as a clear sticky gum. MS: [M+H] + 498.

[0215] Compound 50. 5-[(5-chloro-indan-1-carbonyl)-amino]-4-cyano-3-methyl-thiophene-2-carboxylic acid amide A mixture of 6-chloro-indan-1-carboxylic acid (125 mg, 0.64 mmol) in 2M oxalyl chloride in dichloromethane (0.32 mL, 0.64 mmol) was treated with DMF (2 drops). The reaction was stirred for 2 h and then concentrated. The residue was treated with pyridine (0.35 mL), 5-amino-4-cyano-3-methyl-thiophene-2-carboxylic acid amide (58 mg, 0.32 mmol), and DMAP (5 mg). The mixture was stirred for 16 h. The mixture was treated with water and then extracted with ethyl acetate (2×20 mL). The combined organic extracts were washed with water (2×10 mL) and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by column on silica (0-100% ethyl acetate:hexanes) to give 5-[(5-chloro-indan-1-carbonyl)-amino]-4-cyano-3-methyl-thiophene-2-carboxylic acid amide (101 mg, 88%) as a white solid. MS: [M+H] +360.

[0216] Compound 57. 5-(2-{bis[(4-methoxyphenyl)methyl]amino}-3-phenylpropanamido)-4-cyano-3-methylthiophene-2-carboxamide tert-Butyl 2-amino-3-phenylpropanoate. HCl (600 mg, 2.32 mmol), 1-(bromomethyl)-4-methoxybenzene (0.85 mL, 5.8 mmol), and DIEA (1.4 mL, 8.1 mmol) were taken up in 4 mL of DMF and stirred at room temperature overnight. EtOAc was added and washed with saturated NaHCO3 then with brine. The solvent was removed under vacuum and then purified by normal phase chromatography (0-30% EtOAc / Hex). 900 mg (84% yield) was produced.

[0217] tert-Butyl 2-(bis(4-methoxybenzyl)amino)-3-phenylpropanoate (900 mg, 2.0 mmol) was taken up in 3 mL of DCM and 3 mL of TFA. The reaction was stirred at room temperature overnight. The solvent was removed in vacuo and taken up in EtOAc. The organic layer was washed with saturated NaHCO3 and then with brine. The solvent was removed in vacuo to give 740 mg of 2-(bis(4-methoxybenzyl)amino)-3-phenylpropanoic acid (335 mg, 0.83 mmol), which was treated with fluoro-N,N,N',N'-tetramethylformamidinium hexafluorophosphate (219 mg, 0.83 mmol) and triethylamine (0.58 mL, 4.2 mmol) in 4 mL of DCM at room temperature for 1 h. The solvent was removed in vacuo and taken up in 3 mL of pyridine. 5-Amino-4-cyano-3-methylthiophene-2-carboxamide (50 mg, 0.28 mmol) and a catalytic amount of DMAP were added and the reaction was stirred overnight. EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by normal phase chromatography (0-10% MeOH / DCM). 85 mg (53% yield). MS [M+H] + = 569.

[0218] Compound 68. N-(4-cyano-3-methyl-isothiazol-5-yl)-2-phenyl-butyramide A mixture of 5-amino-3-methyl-isothiazole-4-carbonitrile (50 mg, 0.36 mmol) and DMAP (5 mg) in pyridine (0.5 mL) was treated with 2-phenyl-butyryl chloride (66 mg, 0.36 mmol). The mixture was stirred for 16 h. The mixture was treated with ethyl acetate (30 mL), then washed with 1N HCl (2×20 mL), water (10 mL), and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by column on silica (0-100% ethyl acetate:hexanes) to give N-(4-cyano-3-methyl-isothiazol-5-yl)-2-phenyl-butyramide (60 mg, 59%) as a white solid. MS: [M+H] + 286.

[0219] Compound 75. 5-(2-(4-nitrophenyl)butanamido)-4-cyano-3-methylthiophene-2-carboxamide 2-(4-Nitrophenyl)butanoic acid (0.20 g, 1.0 mmol) was taken up in 5 mL of dry DCM under N2 atmosphere. 1.2 eq of oxalyl chloride (0.10 mL, 1.2 mmol) was added to the solution along with one drop of dry DMF. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The acyl chloride was taken up in 3 mL of pyridine and 5-amino-4-cyano-3-methylthiophene-2-carboxamide (0.14 g, 0.80 mmol) was added. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, followed by washing with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by normal phase chromatography (0-10% MeOH / DCM). 95 mg (32% yield) of solid was produced. MS [M+H] + = 373.

[0220] Compound 82. Methyl 5-(2-phenylbutanamido)-4-cyanothiophene-2-carboxylate Methyl 5-amino-4-cyanothiophene-2-carboxylate (100 mg, 0.55 mmol) was taken up in 3 mL of pyridine. 2-Phenylbutanoyl chloride (118 μL, 0.72 mmol) was added along with a catalytic amount of DMAP. The reaction was stirred overnight. EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by HPLC. 75 mg (43% yield) of methyl 5-(2-phenylbutanamido)-4-cyanothiophene-2-carboxylate was produced as a solid TFA salt. MS [M+H] + = 329.

[0221] Compound 87. Indan-1-carboxylic acid (4-cyano-3-methyl-isothiazol-5-yl)-amide A mixture of indan-1-carboxylic acid (26 mg, 0.16 mmol), 5-amino-3-methyl-isothiazole-4-carbonitrile (45 mg, 0.32 mmol), and trimethylamine (50 mg, 0.48 mmol) in ethyl acetate (1.5 mL) was treated with a 50% solution of cyclic 1-propanephosphonic anhydride in ethyl acetate (0.19 mL, 0.32 mmol). The reaction was heated in a microwave reactor at 160° C. for 15 min. The reaction was diluted with ethyl acetate (15 mL) and then washed with saturated aqueous sodium bicarbonate (10 mL), water (5 mL), and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by preparative HPLC to give indan-1-carboxylic acid (4-cyano-3-methyl-isothiazol-5-yl)-amide as a light yellow solid as the TFA salt (40 mg, 52%). MS: [M+H] + 284.

[0222] Compound 89. 3-Methyl-5-(2-phenyl-butyrylamino)-isothiazole-4-carboxylic acid methyl ester A mixture of 5-amino-3-methyl-isothiazole-4-carboxylic acid methyl ester (67 mg, 0.36 mmol) and DMAP (5 mg) in pyridine (0.5 mL) was treated with 2-phenyl-butyryl chloride (66 mg, 0.36 mmol). The mixture was stirred for 16 h. The mixture was treated with ethyl acetate (30 mL), then washed with 1N HCl (2×10 mL), water (5 mL), and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by column on silica (0-50% ethyl acetate:hexanes). The nearly pure material was dissolved in methanol (1 mL) and eluted with 6N NaOH (50 μL). The reaction was stirred for 2 h and then concentrated. The residue was treated with ethyl acetate (20 mL) and then washed with saturated aqueous sodium carbonate (10 mL), water (5 mL), and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by preparative HPLC to give 3-methyl-5-(2-phenyl-butyrylamino)-isothiazole-4-carboxylic acid methyl ester (8 mg, 7%) as a beige solid. MS: [M+H] + 319.

[0223] Compound 90. N-(4-cyano-isothiazol-5-yl)-2-phenyl-butyramide A mixture of 5-amino-isothiazole-4-carbonitrile (45 mg, 0.36 mmol) and DMAP (5 mg) in pyridine (0.5 mL) was treated with 2-phenyl-butyryl chloride (66 mg, 0.36 mmol). The mixture was stirred for 16 h. The mixture was treated with ethyl acetate (30 mL), washed with 1N HCl (2×20 mL), water (10 mL), and brine (1 mL), dried (Na2SO4), and concentrated. The crude material was purified by column on silica (0-100% ethyl acetate:hexanes) to give N-(4-cyano-isothiazol-5-yl)-2-phenyl-butyramide (81 mg, 83%) as a white solid. MS: [M+H] + 272.

[0224] Compound 95. 2-(2-(2-methoxyphenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate methyl 2-(2-Methoxyphenyl)acetic acid (0.70 g, 4.2 mmol) was taken up in 16 mL of dry THF. The solution was cooled to -78 °C in a dry ice / acetone bath. 2.1 eq of 2.0 M n-butyllithium in hexanes (4.4 mL, 0.010 mol) was added dropwise. The solution was stirred at 0 °C for 2 h. 1.5 eq of iodoethane (0.50 mL, 6.3 mmol) was added slowly and the reaction was stirred at room temperature overnight. The reaction was quenched with water and the volatiles were removed under vacuum. 1N HCl was added to the solution and the product was extracted twice with diethyl ether. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM). 420 mg (51% yield) of a white solid was produced.

[0225] 2-(2-Methoxyphenyl)butanoic acid (70 mg, 0.36 mmol) was taken up in 1 mL of dry DCM under N2 atmosphere. 1.2 eq of oxalyl chloride (34 μL, 0.4 mmol) was added to the solution along with one drop of dry DMF. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The acyl chloride was taken up in 2 mL of pyridine and methyl 2-amino-5-carbamoyl-4-methylthiophene-3-carboxylate (40 mg, 0.18 mmol) was added. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by HPLC. 5 mg (8% yield) of methyl 2-(2-(2-methoxyphenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate was produced as a solid TFA salt. MS [M+H] + = 391.

[0226] Compound 96. 2-(2-(3-methoxyphenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate methyl 2-(3-Methoxyphenyl)acetic acid (0.70 g, 4.2 mmol) was taken up in 16 mL of dry THF. The solution was cooled to -78 °C in a dry ice / acetone bath. 2.1 eq of 2.0 M n-butyllithium in hexanes (4.4 mL, 0.010 mol) was added dropwise. The solution was stirred at 0 °C for 2 h. 1.5 eq of iodoethane (0.50 mL, 6.3 mmol) was added slowly and the reaction was stirred at room temperature overnight. The reaction was quenched with water and the volatiles were removed under vacuum. 1N HCl was added to the solution and the product was extracted twice with diethyl ether. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM). 420 mg (51% yield) of 2-(3-methoxyphenyl)butanoic acid was produced as a white solid.

[0227] 2-(3-Methoxyphenyl)butanoic acid (70 mg, 0.36 mmol) was taken up in 1 mL of dry DCM under N2 atmosphere. 1.2 eq of oxalyl chloride (34 μL, 0.4 mmol) was added to the solution along with one drop of dry DMF. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The acyl chloride was taken up in 2 mL of pyridine and 2-amino-5-carbamoyl-4-methylthiophene-3-carboxylate (40 mg, 0.18 mmol) was added. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified to give 28 mg (44% yield) of 2-(2-(3-methoxyphenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate as a solid TFA salt. MS [M+H] + = 391.

[0228] Compound 97. Methyl 2-(2-(4-methoxyphenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate 2-(4-Methoxyphenyl)butanoic acid (70 mg, 0.36 mmol) was taken up in 1 mL of dry DCM under N2 atmosphere. 1.2 eq of oxalyl chloride (34 μL, 0.4 mmol) was added to the solution along with one drop of dry DMF. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The acyl chloride was taken up in 2 mL of pyridine and methyl 2-amino-5-carbamoyl-4-methylthiophene-3-carboxylate (40 mg, 0.18 mmol) was added. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by HPLC. 14 mg (22% yield) of methyl 2-(2-(4-methoxyphenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate was produced as a solid TFA salt. MS [M+H] + = 391.

[0229] Compound 98. 2-(2-(3-fluorophenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate methyl 2-(3-Fluorophenyl)acetic acid (0.50 g, 3.2 mmol) was taken up in 16 mL of dry THF. The solution was cooled to -78 °C in a dry ice / acetone bath. 2.1 eq of 2.0 M n-butyllithium in hexanes (3.4 mL, 0.067 mol) was added dropwise. The solution was stirred at 0 °C for 2 h. 1.5 eq of iodoethane (0.39 mL, 4.8 mmol) was added slowly and the reaction was stirred at room temperature overnight. The reaction was quenched with water and the volatiles were removed under vacuum. 1N HCl was added to the solution and the product was extracted twice with diethyl ether. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM). 310 mg (53% yield) of 2-(3-fluorophenyl)butanoic acid was produced as a solid.

[0230] 2-(3-Fluorophenyl)butanoic acid (68 mg, 0.36 mmol) was taken up in 1 mL of dry DCM under N2 atmosphere. 1.2 eq of oxalyl chloride (34 μL, 0.4 mmol) was added to the solution along with one drop of dry DMF. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The acyl chloride was taken up in 2 mL of pyridine and methyl 2-amino-5-carbamoyl-4-methylthiophene-3-carboxylate (40 mg, 0.18 mmol) was added. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by HPLC. 14 mg (22% yield) of methyl 2-(2-(3-fluorophenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate was produced as a solid TFA salt. MS [M+H] + = 379.

[0231] Compound 99. 5-Carbamoyl-2-[2-(4-fluoro-phenyl)-butyrylamino]-4-methyl-thiophene-3-carboxylic acid methyl ester 2-(4-Fluorophenyl)acetic acid (6.16 g, 0.040 mol) was taken up in 160 mL of dry THF. The solution was cooled to -78 °C in a dry ice / acetone bath. 2.1 eq of 2.5 M n-butyllithium in hexanes (33.6 mL, 0.084 mol) was added dropwise. The solution was stirred at 0 °C for 2 h. 1.2 eq of iodoethane (3.8 mL, 0.048 mol) was added slowly and the reaction was stirred at room temperature overnight. The reaction was quenched with 40 mL of water and the volatiles were removed under vacuum. 1N HCl was added to the solution and the product was extracted twice with diethyl ether. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM). 5.8 g (80% yield) of 2-(4-fluorophenyl)butanoic acid was produced as a white solid.

[0232] 2-(4-Fluorophenyl)butanoic acid (2.55 g, 0.014 mol) was taken up in 50 mL dry DCM under N2 atmosphere. 1.1 eq oxalyl chloride (1.32 mL, 0.0154 mol) was added to the solution along with 4 drops of dry DMF. The reaction was stirred at room temperature for 1 h. It was quenched with 1 drop of MeOH and the reaction was complete as observed by LC / MS with only the methyl ester. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The product, 2-(4-fluorophenyl)butanoyl chloride, was used as is.

[0233] 2-(4-fluorophenyl)butanoyl chloride (2.8 g, 0.014 mol) and 2-amino-5-carbamoyl-4-methylthiophene-3-carboxylic acid methyl ester (2.35 g, 0.011 mol) were taken up in 50 mL of pyridine. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. 300 mL of EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by normal phase chromatography (0-50% EtOAc / DCM). 1.6 g (30% yield) of 5-carbamoyl-2-[2-(4-fluoro-phenyl)-butyrylamino]-4-methyl-thiophene-3-carboxylic acid methyl ester was produced as a brown solid.

[0234] Compound 106. 5-(2-phenyl-butyrylamino)-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester Ethyl 5-amino-1,2,3-thiadiazole-4-carboxylate (0.169 g, 0.98 mmol) was dissolved in pyridine (5 mL) and then 2-phenylbutyryl chloride (0.196 g, 1.07 mmol) was added. The mixture was stirred at room temperature for 1 h, then diluted with water and extracted with dichloromethane. The extract was concentrated and subjected to chromatography (12 g column, 10-40% ethyl acetate in hexane) to give a white solid (172 mg). MS: [M+H] + 320.05.

[0235] Compound 109. 2-Phenyl-N-[4-(4-trifluoromethyl-phenyl)-[1,2,3]thiadiazol-5-yl]-butyramide 4-(4-Trifluoromethyl-phenyl)-[1,2,3]thiadiazol-5-ylamine (0.073 g, 0.30 mmol) was dissolved in a mixture of dichloromethane (2 mL) and pyridine (0.5 mL). 2-Phenylbutyryl chloride (0.065 g, 0.36 mmol) was added and the mixture was stirred at room temperature for 1 h. The reaction was diluted with water and extracted with dichloromethane. The extract was concentrated and chromatographed (12 g column, 0-30% ethyl acetate in hexanes) to give a white foam (48 mg). MS: [M+H] + 392.

[0236] Compound 110. 2-(2-phenylbutanamido)-N-(4-chlorobenzyl)thiophene-3-carboxamide Methyl 2-(2-phenylbutanamido)thiophene-3-carboxylate (0.45 g, 1.5 mmol) was taken up in 15 mL of MeOH / THF / H2O (1 / 1 / 1) and lithium hydroxide (623 mg, 15 mmol) was added. The reaction was stirred at room temperature overnight. Volatiles were removed under vacuum and the solution was acidified with 4 mL of 1N HCl. The precipitate was filtered off and 320 mg of the resulting 2-(2-phenylbutanamido)thiophene-3-carboxylic acid (50 mg, 0.17 mmol) was treated with 4-chlorobenzylamine (42 μL, 0.34 mmol), EDCI (67 mg, 0.34 mmol), HOBt (53 mg, 0.34 mmol), and DIEA (89 μL, 0.51 mmol) in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give 2-(2-phenylbutanamido)-N-(4-chlorobenzyl)thiophene-3-carboxamide (45 mg, 64% yield) as the TFA salt. MS [M+H] + = 413.

[0237] Compound 111. 4-Pyridin-2-yl-piperazine-1-carboxylic acid [3-(4-chloro-benzylcarbamoyl)-thiophen-2-yl]-amide Methyl 2-aminothiophene-3-carboxylate (2.47 g, 0.016 mol) and phenyl chloroformate (2.4 mL, 0.019 mol) were taken up in 70 mL of dry THF under N2 atmosphere. Pyridine (1.9 mL, 0.023) and a catalytic amount of DMAP were added and the reaction was stirred at room temperature overnight. 300 mL of EtOAc was added, followed by washing with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum. 4.35 g (quantitative) of phenyl 3-(methoxycarbonyl)thiophen-2-ylcarbamate was produced, which was used as is.

[0238] Phenyl 3-(methoxycarbonyl)thiophen-2-ylcarbamate (4.35 g, 0.016 mol) and 1-(pyridin-2-yl)piperazine (2.95 mL, 0.020 mol) were taken up in 60 mL of DMF. DIEA (6.4 mL, 0.037 mol) was added and the reaction was stirred at room temperature overnight. EtOAc was added and washed with saturated NaHCO3 and brine. The crude material was purified by normal phase chromatography (0-10% MeOH / DCM). 4.8 g (86% yield) of a solid, methyl 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)thiophene-3-carboxylate was produced.

[0239] Methyl 2-(4-(pyridin-2-yl)piperazine-1-carboxamido)thiophene-3-carboxylate (4.75 g, 0.014 mol) was taken up in 60 mL of MeOH / THF / H2O (1 / 1 / 1) and LiOH·H2O (5.8 g, 0.14 mol) was added. The reaction was stirred overnight at room temperature. The volatiles were removed under vacuum and 150 mL of 1N HCl was added. The product, 2-(4-(pyridin-2-yl)piperazine-1-carboxamido)thiophene-3-carboxylic acid, precipitated out of solution, was filtered and washed with water (3.5 g, 77% yield).

[0240] 2-(4-(pyridin-2-yl)piperazine-1-carboxamido)thiophene-3-carboxylic acid (3.0 g, 9.04 mmol), 4-chlorobenzylamine (1.21 mL, 9.9 mmol), EDCI (2.1 g, 10.8 mmol), HOBt (1.65 g, 10.8 mmol), and DIEA (3.15 mL, 18 mmol) were taken in 35 mL of DMF and stirred at room temperature overnight. EtOAc was added and washed with saturated NaHCO3 and brine. The crude material was purified by normal phase chromatography (0-10% MeOH / DCM). 3.0 g (73% yield) was produced. The solid was converted to the HCl salt by adding 3.5 mL of 4N HCl in dioxane to the material in 5 mL of dioxane. The solution was stirred for 4 h and then frozen. The solvent was removed by lyophilization to give 3.25 g of 4-pyridin-2-yl-piperazine-1-carboxylic acid [3-(4-chloro-benzylcarbamoyl)-thiophen-2-yl]-amide.

[0241] Compound 112. Methyl 2-(2-phenylbutanamido)thiophene-3-carboxylate Methyl 2-aminothiophene-3-carboxylate (250 mg, 1.6 mmol) was taken up in 8 mL of pyridine. 2-Phenylbutanoyl chloride (221 μL, 1.6 mmol) was added along with a catalytic amount of DMAP. The reaction was stirred overnight. EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by normal phase chromatography. 450 mg (93% yield) of methyl 2-(2-phenylbutanamido)thiophene-3-carboxylate was produced as a solid. MS [M+H] + = 304.

[0242] Compound 114. Methyl 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)thiophene-3-carboxylate Methyl 2-aminothiophene-3-carboxylate (2.47 g, 0.016 mol) and phenyl chloroformate (2.4 mL, 0.019 mol) were taken up in 70 mL of dry THF under N2 atmosphere. Pyridine (1.9 mL, 0.023) and a catalytic amount of DMAP were added and the reaction was stirred at room temperature overnight. EtOAc (300 mL) was added and the solution was washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed in vacuo to give phenyl 3-(methoxycarbonyl)thiophen-2-ylcarbamate (4.35 g, quantitative), which was used as is.

[0243] Phenyl 3-(methoxycarbonyl)thiophen-2-ylcarbamate (4.35 g, 0.016 mol) and 1-(pyridin-2-yl)piperazine (2.95 mL, 0.020 mol) were taken up in 60 mL of DMF. DIEA (6.4 mL, 0.037 mol) was added and the reaction was stirred at room temperature overnight. EtOAc was added and washed with saturated NaHCO3 and brine. The crude material was purified by normal phase chromatography (0-10% MeOH / DCM). 4.8 g (86% yield) of methyl 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)thiophene-3-carboxylate was produced as a solid. MS [M+H] + = 347.

[0244] Compound 115. 2-(2-phenylbutanamido)-N3-(4-chlorobenzyl)-4-methylthiophene-3,5-dicarboxamide Methyl 2-(2-phenylbutanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate (100 mg, 0.28 mmol) was dissolved in 9 mL of MeOH / THF / H2O (1 / 1 / 1) and lithium hydroxide (116 mg, 2.8 mmol) was added. The reaction was stirred overnight at room temperature. The volatiles were removed in vacuo and the solution was acidified with 4 mL of 1N HCl. The precipitate was filtered off and 60 mg of 2-(2-phenylbutanamido)-5-carbamoyl-4-methylthiophene-3-carboxylic acid was used as is.

[0245] 2-(2-phenylbutanamido)-5-carbamoyl-4-methylthiophene-3-carboxylic acid (30 mg, 0.09 mmol), 4-chlorobenzylamine (15 mg, 0.11 mmol), EDCI (25 mg, 0.14 mmol), HOBt (21 mg, 0.14 mmol), and DIEA (47 μL, 0.23 mmol) were taken in 0.5 mL of DMF and stirred overnight at room temperature. The reaction was purified by preparative HPLC to give 2-(2-phenylbutanamido)-N3-(4-chlorobenzyl)-4-methylthiophene-3,5-dicarboxamide (20 mg, 47% yield) as the TFA salt. MS [M+H] + = 470.

[0246] Compound 116. Methyl 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)-5-carbamoyl-4-methylthiophene-3-carboxylate 2-Amino-5-carbamoyl-4-methylthiophene-3-carboxylic acid (50 mg, 0.23 mmol) was taken up in 3 mL of pyridine. Phenyl chloroformate (95 μL, 0.75 mmol) was added along with a catalytic amount of DMAP. The reaction was stirred overnight. EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by normal phase chromatography (0-10% MeOH / DCM) to give phenyl 3-(methoxycarbonyl)-5-carbamoyl-4-methylthiophen-2-ylcarbamate (77 mg, quantitative yield).

[0247] Phenyl 3-(methoxycarbonyl)-5-carbamoyl-4-methylthiophen-2-ylcarbamate (77 mg, 0.23 mmol), 1-(pyridin-2-yl)piperazine (67 μL, 0.46 mmol), and triethylamine (120 μL, 0.69 mmol) were taken up in 1 mL of DMF and stirred at room temperature overnight. EtOAc was added, then washed with saturated sodium bicarbonate and brine. The solvent was removed under vacuum and purified by normal phase chromatography (0-10% MeOH / DCM) to give methyl 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)-5-carbamoyl-4-methylthiophene-3-carboxylate (85 mg, 90% yield). MS [M+H] + = 404.

[0248] Compound 118. 4-Pyridin-2-yl-piperazine-1-carboxylic acid [4-(4-trifluoromethyl-phenyl)-[1,2,3]thiadiazol-5-yl]-amide Trifluoromethyl-phenyl)-[1,2,3]thiadiazol-5-ylamine (0.120 g, 0.49 mmol) was dissolved in a mixture of dichloromethane (5 mL) and pyridine (1 mL). Phenyl chloroformate (0.085 g, 0.54 mmol) was added and the mixture was stirred at room temperature for 2 hours. The reaction was diluted with water and extracted with dichloromethane. The extract was concentrated and then dissolved in tetrahydrofuran (2 mL) in a pressure tube. 1-Pyridin-2-yl-piperazine (0.40 g, 0.49 mmol) was added, the tube was sealed, and the mixture was heated to about 80° C. overnight. The reaction mixture was concentrated and the residue was chromatographed (12 g column, 10-40% ethyl acetate in hexanes) to give 4-pyridin-2-yl-piperazine-1-carboxylic acid [4-(4-trifluoromethyl-phenyl)-[1,2,3]thiadiazol-5-yl]-amide as a white solid (52 mg). MS: [M+H] + 434.96.

[0249] Compound 119. 4-Pyridin-2-yl-piperazine-1-carboxylic acid (4-benzylcarbamoyl-[1,2,3]thiadiazol-5-yl)-amide 5-[(4-Pyridin-2-yl-piperazine-1-carbonyl)-amino]-[1,2,3]thiadiazole-4-carboxylic acid (0.046 g, 0.14 mmol) was suspended in anhydrous dimethylformamide. Diisopropylethylamine (0.089 g, 0.69 mmol) was added followed by benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (0.143 g, 0.275 mmol). The mixture was stirred at room temperature for 15 minutes, then benzylamine (0.029 g, 0.275 mmol) was added. The mixture was stirred at room temperature for 16 hours, diluted with brine and extracted with ethyl acetate. The extract was concentrated and chromatographed (12 g column, 25-50% ethyl acetate in hexanes) to give 4-pyridin-2-yl-piperazine-1-carboxylic acid (4-benzylcarbamoyl-[1,2,3]thiadiazol-5-yl)-amide as a white solid (5 mg). MS: [M+H] + 423.96.

[0250] Compound 123. 4-Phenyl-piperazine-1-carboxylic acid (4-benzylcarbamoyl-[1,2,3]thiadiazol-5-yl)-amide 5-Phenoxycarbonylamino-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester (0.331 g, 1.13 mmol), 1-phenylpiperazine hydrochloride (0.247 g, 1.24 mmol), and diisopropylethylamine (0.219 g, 1.70 mmol) were dissolved in anhydrous tetrahydrofuran in a pressure tube. The tube was sealed and heated at approximately 80 °C for 16 h. The reaction was cooled, diluted with saturated aqueous sodium bicarbonate, and extracted with ethyl acetate. The extract was concentrated and subjected to chromatography (12 g column, 20-50% ethyl acetate in hexanes) to give 5-[(4-phenyl-piperazine-1-carbonyl)-amino]-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester (398 mg) as a white solid.

[0251] 5-[(4-Phenyl-piperazine-1-carbonyl)-amino]-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester (0.393 g, 1.09 mmol) was dissolved in tetrahydrofuran (15 mL). Lithium hydroxide hydrate (0.228 g, 5.44 mmol) was added, followed by approximately 4 mL of water, enough to dissolve most of the solid. The reaction was stirred at room temperature for 16 h. Aqueous hydrochloric acid (0.1 N) was added and the mixture was extracted repeatedly with ethyl acetate. The extracts were concentrated to a white solid, which was subjected to chromatography (12 g column, 2-10% methanol in dichloromethane) to give 5-[(4-Phenyl-piperazine-1-carbonyl)-amino]-[1,2,3]thiadiazole-4-carboxylic acid (212 mg) as a white solid.

[0252] 5-[(4-phenyl-piperazine-1-carbonyl)-amino]-[1,2,3]thiadiazole-4-carboxylic acid (0.051 g, 0.15 mmol) was suspended in anhydrous dimethylformamide. Diisopropylethylamine (0.040 g, 0.31 mmol) was added followed by benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (0.124 g, 0.23 mmol). The mixture was stirred at room temperature for 15 minutes and then benzylamine (0.025 g, 0.23 mmol) was added. The mixture was stirred at room temperature for 16 hours, diluted with brine and extracted with ethyl acetate. The extract was concentrated and chromatographed (12 g column, 25-50% ethyl acetate in hexanes) to give 4-phenyl-piperazine-1-carboxylic acid (4-benzylcarbamoyl-[1,2,3]thiadiazol-5-yl)-amide as a white solid (27 mg). MS: [M+H] + 422.94.

[0253] Compound 124. 4-Pyridin-2-yl-piperazine-1-carboxylic acid [4-(4-methyl-benzylcarbamoyl)-[1,2,3]thiadiazol-5-yl]-amide 5-[(4-phenyl-piperazine-1-carbonyl)-amino]-[1,2,3]thiadiazole-4-carboxylic acid (0.051 g, 0.15 mmol) was suspended in anhydrous dimethylformamide. Diisopropylethylamine (0.040 g, 0.31 mmol) was added followed by benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (0.124 g, 0.23 mmol). The mixture was stirred at room temperature for 15 minutes and then 4-methylbenzylamine (0.028 g, 0.23 mmol) was added. The mixture was stirred at room temperature for 16 hours, diluted with brine and extracted with ethyl acetate. The extract was concentrated and chromatographed (12 g column, 25-50% ethyl acetate in hexanes) to give 4-pyridin-2-yl-piperazine-1-carboxylic acid [4-(4-methyl-benzylcarbamoyl)-[1,2,3]thiadiazol-5-yl]-amide as a white solid (31 mg) MS: [M+H] + 437.96.

[0254] Compound 127. Ethyl 5-(4-(pyridin-2-yl)piperazine-1-carboxamide)thiazole-4-carboxylate Ethyl 5-aminothiazole-4-carboxylate (500 mg, 02.9 mmol) was dissolved in 30 mL of THF. Phenyl chloroformate (403 μL, 3.1 mmol) was added along with pyridine (290 μL, 3.5 mmol). The reaction was stirred overnight. EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by normal phase chromatography (0-10% MeOH / DCM). 850 mg (quantitative yield) of a solid was produced.

[0255] Phenyl 4-(ethoxycarbonyl)thiazol-5-ylcarbamate (400 mg, 1.3 mmol), 1-(pyridin-2-yl)piperazine (210 μL, 1.4 mmol), and DIEA (450 μL, 2.6 mmol) were taken up in 8 mL of DMF and stirred at room temperature overnight. EtOAc was added, then washed with saturated sodium bicarbonate and brine. The solvent was removed under vacuum and purified by normal phase chromatography (0-10% MeOH / DCM) to give ethyl 5-(4-(pyridin-2-yl)piperazine-1-carboxamide)thiazole-4-carboxylate (430 mg, 92% yield) as a solid. MS [M+H] + = 362.

[0256] Compound 130. N-(3-(benzylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)thiophene-3-carboxylic acid (33 mg, 0.10 mmol), benzylamine (25 μL, 0.11 mmol), EDCI (38 mg, 0.20 mmol), HOBt (38 mg, 0.20 mmol), and DIEA (52 μL, 0.30 mmol) were taken in 0.5 mL of DMF and stirred overnight at room temperature. The reaction was purified by preparative HPLC to give N-(3-(benzylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide (40 mg, 75% yield) as the TFA salt. MS [M+H] + = 422.

[0257] Compound 131. 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)-N3-benzyl-4-methylthiophene-3,5-dicarboxamide 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)-5-carbamoyl-4-methylthiophene-3-carboxylic acid (38 mg, 0.10 mmol), benzylamine (25 μL, 0.11 mmol), EDCI (38 mg, 0.20 mmol), HOBt (38 mg, 0.20 mmol), and DIEA (52 μL, 0.30 mmol) were taken in 0.5 mL of DMF and stirred overnight at room temperature. The reaction was purified by preparative HPLC to give 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)-N3-benzyl-4-methylthiophene-3,5-dicarboxamide (44 mg, 74% yield) as the TFA salt. MS [M+H] + = 479.

[0258] Compound 132. [4-(4-chloro-benzylcarbamoyl)-[1,2,3]thiadiazol-5-yl]-carbamic acid phenyl ester Ethyl 5-amino-1,2,3-thiadiazole-4-carboxylate (0.328 g, 1.89 mmol) and triethylamine (0.287 g, 2.84 mmol) were dissolved in anhydrous dichloromethane (25 ml). Di-tert-butyl dicarbonate (0.413 g, 1.89 mmol) was added and the reaction was then stirred at room temperature for 16 hours. The mixture was diluted with saturated aqueous sodium bicarbonate and extracted with dichloromethane. The extract was concentrated and subjected to chromatography (20 g column, 10-50% ethyl acetate in hexanes) to give 5-tert-butoxycarbonylamino-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester (352 mg).

[0259] 5-tert-Butoxycarbonylamino-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester (0.352 g, 1.29 mmol) and lithium hydroxide hydrate (0.162 g, 3.86 mmol) were mixed in tetrahydrofuran (40 mL). Sufficient water was added to bring most of the solid into solution (8 mL). The mixture was stirred at room temperature overnight. TLC indicated the reaction was incomplete, so it was heated to reflux for 24 h. The reaction was then acidified with 0.1 N aqueous hydrochloric acid and extracted with ethyl acetate. The extract was concentrated and subjected to chromatography (20 g column, 2-10% methanol in dichloromethane) to give 5-tert-butoxycarbonylamino-[1,2,3]thiadiazole-4-carboxylic acid (270 mg).

[0260] 5-tert-Butoxycarbonylamino-[1,2,3]thiadiazole-4-carboxylic acid (0.270 g, 1.10 mmol) and diisopropylethylamine (0.284 g, 2.20 mmol) were dissolved in anhydrous dimethylformamide (10 mL). Benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (0.859 g, 1.65 mmol) was added and the mixture was stirred at room temperature for 15 minutes. 4-Chlorobenzylamine (0.234 g, 1.65 mmol) was added. The mixture was stirred at room temperature for 16 hours, diluted with brine and extracted with ethyl acetate. The extract was concentrated and chromatographed (40 g column, 100% dichloromethane) to give a white solid, [4-(4-chloro-benzylcarbamoyl)-[1,2,3]thiadiazol-5-yl]-carbamic acid tert-butyl ester (404 mg).

[0261] [4-(4-chloro-benzylcarbamoyl)-[1,2,3]thiadiazol-5-yl]-carbamic acid tert-butyl ester (0.404 g, 1.10 mmol) was dissolved in a mixture of dichloromethane (25 mL) and trifluoroacetic acid (5 mL) and the solution was stirred at room temperature for 16 h. The reaction was carefully basified with saturated aqueous sodium bicarbonate and then extracted with dichloromethane. The extract was concentrated and subjected to chromatography (40 g column, 0-5% methanol in dichloromethane) to give 5-amino-[1,2,3]thiadiazole-4-carboxylic acid 4-chloro-benzylamide (291 mg) as a white solid.

[0262] 5-Amino-[1,2,3]thiadiazole-4-carboxylic acid 4-chloro-benzylamide (0.291 g, 1.08 mmol) and triethylamine (0.164 g, 1.19 mmol) were dissolved in anhydrous dichloromethane. Phenyl chloroformate (0.187 g, 1.19 mmol) was added and the mixture was stirred at room temperature for 6 h. Saturated aqueous sodium bicarbonate was added and the mixture was extracted with dichloromethane. The extract was concentrated and subjected to chromatography (20 g column, 10-50% ethyl acetate in hexane) to give the solid [4-(4-chloro-benzylcarbamoyl)-[1,2,3]thiadiazol-5-yl]-carbamic acid phenyl ester (351 mg).

[0263] Compound 134. 5-(2-phenyl-butyrylamino)-[1,2,3]thiadiazole-4-carboxylic acid 4-chloro-benzylamide 5-(2-Phenyl-butyrylamino)-[1,2,3]thiadiazole-4-carboxylic acid (0.073 g, 0.25 mmol) was suspended in anhydrous dimethylformamide (3 mL). Diisopropylethylamine (0.065 g, 0.50 mmol) was added, followed by benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (0.196 g, 0.38 mmol). The mixture was stirred at room temperature for 15 min, then 4-chlorobenzylamine (0.040 g, 0.38 mmol) was added. The mixture was stirred at room temperature for 16 h, diluted with brine, and extracted with ethyl acetate. The extract was concentrated and subjected to chromatography (12 g column, 20-50% ethyl acetate in hexanes) to give 5-(2-Phenyl-butyrylamino)-[1,2,3]thiadiazole-4-carboxylic acid 4-chloro-benzylamide as a white solid (36 mg). MS: [M+H] + 415.87.

[0264] Compound 135. 5-(2-phenyl-butyrylamino)-[1,2,3]thiadiazole-4-carboxylic acid benzylamide Ethyl 5-amino-1,2,3-thiadiazole-4-carboxylate (0.169 g, 0.98 mmol) was dissolved in pyridine (5 mL). 2-Phenylbutyryl chloride (0.196 g, 1.07 mmol) was added. The mixture was stirred at room temperature for 1 h, diluted with water, and extracted with dichloromethane. The extract was concentrated and subjected to chromatography (12 g column, 10-40% ethyl acetate in hexanes) to give 5-(2-phenyl-butyrylamino)-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester (172 mg) as a white solid.

[0265] 5-(2-Phenyl-butyrylamino)-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester (0.167 g, 0.52 mmol) was dissolved in tetrahydrofuran. Lithium hydroxide hydrate (0.110 g, 2.61 mmol) was added, followed by enough water to bring most of the solid into solution (4 mL). The mixture was heated at reflux for 16 h, cooled to room temperature, and acidified with 0.1 N hydrochloric acid. The mixture was extracted with ethyl acetate. The extract was concentrated and chromatographed (12 g column, 2-10% methanol in dichloromethane) to give 5-(2-phenyl-butyrylamino)-[1,2,3]thiadiazole-4-carboxylic acid (151 mg) as a white solid.

[0266] 5-(2-Phenyl-butyrylamino)-[1,2,3]thiadiazole-4-carboxylic acid (0.073 g, 0.25 mmol) was suspended in anhydrous dimethylformamide (3 mL). Diisopropylethylamine (0.065 g, 0.50 mmol) was added, followed by benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (0.196 g, 0.38 mmol). The mixture was stirred at room temperature for 15 min, then benzylamine (0.040 g, 0.38 mmol) was added. The mixture was stirred at room temperature for 16 h, diluted with brine, and extracted with ethyl acetate. The extract was concentrated and subjected to chromatography (12 g column, 20-50% ethyl acetate in hexane) to give 5-(2-Phenyl-butyrylamino)-[1,2,3]thiadiazole-4-carboxylic acid benzylamide as a white solid (28 mg). MS: [M+H] + 381.92.

[0267] Compound 144. N-(3-((6-chloropyridin-2-yl)methylcarbamoyl)thiophen-2-yl)-4-phenylpiperazine-1-carboxamide 2-(4-phenylpiperazine-1-carboxamide)thiophene-3-carboxylic acid (33 mg, 0.1 mmol), (6-chloropyridin-3-yl)methanamine (28 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred overnight at room temperature. The reaction was purified by preparative HPLC to give N-(3-((6-chloropyridin-2-yl)methylcarbamoyl)thiophen-2-yl)-4-phenylpiperazine-1-carboxamide (25 mg, 55% yield). MS [M+H] + = 456.

[0268] Compound 153. N-[4-(2-fluoro-phenyl)-[1,2,3]thiadiazol-5-yl]-2-phenyl-butyramide 4-(2-Fluorophenyl)thiadiazol-5-amine (0.050 g, 0.26 mmol) and triethylamine (0.039 g, 0.39 mmol) were dissolved in anhydrous dichloromethane (2 mL). 2-Phenylbutyryl chloride (0.056 g, 0.31 mmol) was added. The reaction was stirred at room temperature for 3 h. The reaction was diluted with saturated aqueous sodium bicarbonate and extracted with dichloromethane. The extract was concentrated and chromatographed (12 g column, 10-50% ethyl acetate in hexanes) to give the product as a solid (4 mg). MS: [M+H] + 342.05.

[0269] Compound 154. N-[4-(4-fluoro-phenyl)-[1,2,3]thiadiazol-5-yl]-2-phenyl-butyramide 4-(4-Fluorophenyl)thiadiazol-5-amine (0.050 g, 0.26 mmol) and triethylamine (0.039 g, 0.39 mmol) were dissolved in anhydrous dichloromethane (2 mL). 2-Phenylbutyryl chloride (0.056 g, 0.31 mmol) was added. The reaction was stirred at room temperature for 3 h. The reaction was diluted with saturated aqueous sodium bicarbonate and extracted with dichloromethane. The extract was concentrated and chromatographed (12 g column, 10-50% ethyl acetate in hexanes) to give the product as a solid (46 mg). MS: [M+H] + 342.05.

[0270] Compound 155. 2-Phenyl-N-(4-phenyl-[1,2,3]thiadiazol-5-yl)-butyramide 4-Phenylthiadiazol-5-amine (0.045 g, 0.26 mmol) and triethylamine (0.039 g, 0.39 mmol) were dissolved in anhydrous dichloromethane (2 mL). 2-Phenylbutyryl chloride (0.056 g, 0.31 mmol) was added. The reaction was stirred at room temperature for 3 h. The reaction was diluted with saturated aqueous sodium bicarbonate and extracted with dichloromethane. The extract was concentrated and chromatographed (12 g column, 10-50% ethyl acetate in hexanes) to give the product as a solid (12 mg). MS: [M+H] + 324.06.

[0271] Compound 159. N-(3-(3,4-difluorobenzylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)thiophene-3-carboxylic acid (33 mg, 0.1 mmol), (3,4-difluorophenyl)methanamine (28 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give N-(3-(3,4-difluorobenzylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide (41 mg, 89% yield) as the TFA salt. MS [M+H] + = 458.

[0272] Compound 160. 2-(2-phenylbutanamido)-N-(3,4-difluorobenzyl)thiophene-3-carboxamide 2-(2-phenylbutanamido)thiophene-3-carboxylic acid (29 mg, 0.1 mmol), (3,4-difluorophenyl)methanamine (28 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken up in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give 2-(2-phenylbutanamido)-N-(3,4-difluorobenzyl)thiophene-3-carboxamide (37 mg, 89% yield) as the TFA salt. MS [M+H] + = 415.

[0273] Compound 161. 2-(2-phenylbutanamido)-N-benzylthiophene-3-carboxamide 2-(2-phenylbutanamido)thiophene-3-carboxylic acid (29 mg, 0.1 mmol), benzylamine (21 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken up in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give 2-(2-phenylbutanamido)-N-benzylthiophene-3-carboxamide (28 mg, 75% yield) as the TFA salt. MS [M+H]+ = 379.

[0274] Compound 162. 5-(2-phenylbutanamido)-N-benzylthiazole-4-carboxamide 5-(2-phenylbutanamido)thiazole-4-carboxylic acid (29 mg, 0.1 mmol), benzylamine (21 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken up in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by chromatography to give 5-(2-phenylbutanamido)-N-benzylthiazole-4-carboxamide (12 mg, 31% yield). MS [M+H] + = 380.

[0275] Compound 163. 5-(2-phenylbutanamido)-N-(4-chlorobenzyl)thiazole-4-carboxamide Ethyl 5-(2-phenylbutanamido)thiazole-4-carboxylate (140 mg, 0.44 mmol) was taken up in 5 mL of MeOH / H2O (1 / 1) and lithium hydroxide (184 mg, 4.4 mmol) was added. The reaction was stirred at room temperature overnight. The volatiles were removed in vacuo and the solution was acidified with 4 mL of 1N HCl. The precipitate was filtered off and 100 mg of the product was used as is.

[0276] 5-(2-phenylbutanamido)thiazole-4-carboxylic acid (29 mg, 0.1 mmol), 4-chlorobenzylamine (28 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give 5-(2-phenylbutanamido)-N-(4-chlorobenzyl)thiazole-4-carboxamide (13 mg, 31% yield) as the TFA salt. MS [M+H] + = 414.

[0277] Compound 165. Ethyl 5-(2-phenylbutanamido)thiazole-4-carboxylate Ethyl 5-aminothiazole-4-carboxylate (250 mg, 1.45 mmol), 2-phenylbutanoyl chloride (265 μL, 1.6 mmol), and pyridine (140 μL, 1.8 mmol) were taken up in 5 mL of THF. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, then washed with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purified by normal phase chromatography to give ethyl 5-(2-phenylbutanamido)thiazole-4-carboxylate (140 mg, 30% yield) as a solid. MS [M+H] + = 319.

[0278] Compound 167. N-(4-(4-chlorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-1-(pyridin-2-yl)piperidine-4-carboxamide N-(4-chlorobenzyl)-5-amino-1,2,3-thiadiazole-4-carboxamide (25 mg, 0.1 mmol), 1-(pyridin-2-yl)piperidine-4-carboxylic acid (21 mg, 0.10 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred overnight at room temperature. The reaction was purified by chromatography to give N-(4-(4-chlorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-1-(pyridin-2-yl)piperidine-4-carboxamide (8 mg, 18% yield). MS [M+H] + = 457.

[0279] Compound 168. 4-(2-Fluoro-phenyl)-piperazine-1-carboxylic acid [4-(4-chloro-benzylcarbamoyl)-[1,2,3]thiadiazol-5-yl]-amide 5-Phenoxycarbonylamino-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester (0.177 g, 0.603 mmol), 1-(2-fluorophenyl)piperazine hydrochloride (0.144 g, 0.664 mmol), and diisopropylethylamine (0.117 g, 0.905 mmol) were dissolved in anhydrous tetrahydrofuran (5 mL) and the solution was heated to reflux for 16 h. The reaction was cooled to room temperature, concentrated to dryness, and subjected to chromatography on silica gel (24 g column, 20-50% ethyl acetate in hexanes) to give 5-{[4-(2-fluoro-phenyl)-piperazine-1-carbonyl]-amino}-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester (210 mg), a white solid.

[0280] 5-{[4-(2-fluoro-phenyl)-piperazine-1-carbonyl]-amino}-[1,2,3]thiadiazole-4-carboxylic acid ethyl ester (0.210 g, 0.553 mmol) and lithium hydroxide hydrate (0.116 g, 2.77 mmol) were mixed in tetrahydrofuran (20 mL). Water was added until most of the solid went into solution (approximately 5 mL). The reaction was stirred at room temperature for 3 days. Hydrochloric acid (0.1 N) was added and the mixture was repeatedly extracted with ethyl acetate. The extracts were concentrated and subjected to chromatography (12 g column, gradient from 100% dichloromethane to 0.5:10:90 acetic acid:methanol:dichloromethane) to give 5-{[4-(2-fluoro-phenyl)-piperazine-1-carbonyl]-amino}-[1,2,3]thiadiazole-4-carboxylic acid (110 mg) as a white solid.

[0281] 5-{[4-(2-fluoro-phenyl)-piperazine-1-carbonyl]-amino}-[1,2,3]thiadiazole-4-carboxylic acid (0.040 g, 0.11 mmol) was dissolved in anhydrous dimethylformamide (2 mL). Diisopropylethylamine (0.022 g, 0.17 mmol) was added, followed by benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (0.071 g, 0.14 mmol). The mixture was stirred at room temperature for 15 minutes, then 4-chlorobenzylamine (0.019 g, 0.14 mmol) was added. The mixture was stirred at room temperature for 16 hours, diluted with brine, and extracted with ethyl acetate. The extract was concentrated and chromatographed (12 g column, 0-40% ethyl acetate in hexanes) to give 4-(2-fluoro-phenyl)-piperazine-1-carboxylic acid [4-(4-chloro-benzylcarbamoyl)-[1,2,3]thiadiazol-5-yl]-amide as a white solid (13 mg). MS: [M+H] + 475.12.

[0282] Compound 169. 4-(2-Fluoro-phenyl)-piperazine-1-carboxylic acid (4-benzylcarbamoyl-[1,2,3]thiadiazol-5-yl)-amide 5-{[4-(2-fluoro-phenyl)-piperazine-1-carbonyl]-amino}-[1,2,3]thiadiazole-4-carboxylic acid (0.040 g, 0.11 mmol) was dissolved in anhydrous dimethylformamide (2 mL). Diisopropylethylamine (0.022 g, 0.17 mmol) was added, followed by benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (0.071 g, 0.14 mmol). The mixture was stirred at room temperature for 15 minutes, then benzylamine (0.015 g, 0.14 mmol) was added. The mixture was stirred at room temperature for 16 hours, diluted with brine, and extracted with ethyl acetate. The extract was concentrated and chromatographed (12 g column, 0-40% ethyl acetate in hexanes) to give 4-(2-fluoro-phenyl)-piperazine-1-carboxylic acid (4-benzylcarbamoyl-[1,2,3]thiadiazol-5-yl)-amide as a white solid (3 mg). MS: [M+H] + 440.94.

[0283] Compound 170. N-(3-((6-chloropyridin-3-yl)methylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)thiophene-3-carboxylic acid (33 mg, 0.1 mmol), (6-chloropyridin-3-yl)methanamine (28 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC, which yielded 12 mg (21% yield) of the TFA salt of N-(3-((6-chloropyridin-3-yl)methylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide. MS [M+H] + = 457.

[0284] Compound 172. N-(3-(N-(2-methoxybenzyl)-N-methylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)thiophene-3-carboxylic acid (33 mg, 0.1 mmol), (2-methoxyphenyl)-N-methylmethanamine (30 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by chromatography to give N-(3-(N-(2-methoxybenzyl)-N-methylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide (40 mg, 69% yield) as the TFA salt. MS [M+H] + = 466.

[0285] Compound 173. N-(3-(N-benzyl-N-methylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide 2-(4-(pyridin-2-yl)piperazine-1-carboxamide)thiophene-3-carboxylic acid (33 mg, 0.1 mmol), N-methyl(phenyl)methanamine (24 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred overnight at room temperature. The reaction was purified by chromatography to give N-(3-(N-benzyl-N-methylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide (35 mg, 64% yield) as the TFA salt. MS [M+H] + = 436.

[0286] Compound 174. Methyl 5-(2-(4-fluorophenyl)butanoylcarbamoyl)-2-amino-4-methylthiophene-3-carboxylate A small amount of by-product was observed during the preparation of compound 99, which was purified by normal phase chromatography and identified as methyl 5-(2-(4-fluorophenyl)butanoylcarbamoyl)-2-amino-4-methylthiophene-3-carboxylate. MS [M+H] + = 379.

[0287] Compound 175. 5-(4-(pyridin-2-yl)butanamido)-N-(4-chlorobenzyl)-1,2,3-thiadiazole-4-carboxamide N-(4-chlorobenzyl)-5-amino-1,2,3-thiadiazole-4-carboxamide. TFA (40 mg, 0.1 mmol), 4-(pyridin-2-yl)butanoic acid (21 mg, 0.13 mmol), EDCI (29 mg, 0.15 mmol), HOBt (23 mg, 0.15 mmol), and DIEA (52 μL, 0.3 mmol) were taken in 1.0 mL of DMF and stirred overnight at room temperature. The reaction was purified by preparative HPLC to give 5-(4-(pyridin-2-yl)butanamido)-N-(4-chlorobenzyl)-1,2,3-thiadiazole-4-carboxamide (5 mg, 12% yield) as the TFA salt. MS [M+H] + = 416.

[0288] Compound 179. 2-(2-phenylbutanamido)-N-(4-fluorobenzyl)thiophene-3-carboxamide 2-(2-phenylbutanamido)thiophene-3-carboxylic acid (29 mg, 0.10 mmol), 4-fluorobenzylamine (17 μL, 0.15 mmol), EDCI (29 mg, 0.15 mmol), and HOBt (23 mg, 0.15 mmol) were taken in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give 2-(2-phenylbutanamido)-N-(4-fluorobenzyl)thiophene-3-carboxamide (37 mg, 93% yield) as the TFA salt. MS [M+H] + = 397.

[0289] Compound 180. N-(4-(4-fluorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide 5-(4-(pyridin-2-yl)piperazine-1-carboxamide)-1,2,3-thiadiazole-4-carboxylic acid (33 mg, 0.10 mmol), 4-fluorobenzylamine (17 μL, 0.15 mmol), EDCI (29 mg, 0.15 mmol), and HOBt (23 mg, 0.15 mmol) were taken in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give N-(4-(4-fluorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide (40 mg, 70% yield) as the TFA salt. MS [M+H] + = 442.

[0290] Compound 181. N-(3-(4-fluorobenzylcarbamoyl)thiophen-2-yl)-4-(pyridin-2-yl)piperazine-1-carboxamide 2-(4-(pyridin-2-yl)piperazine-1-carboxamido)thiophene-3-carboxylic acid (33 mg, 0.1 mmol), (4-fluorophenyl)methanamine (25 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC. 25 mg (45% yield) of TFA salt was produced. MS [M+H] + = 440.

[0291] Compound 182. 1-(4-(4-fluorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-3-(2-morpholinoethyl)urea 5-(3-(2-morpholinoethyl)ureido)-1,2,3-thiadiazole-4-carboxylic acid (23 mg, 0.08 mmol), (4-fluorophenyl)methanamine (25 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give 1-(4-(4-fluorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-3-(2-morpholinoethyl)urea (21 mg, 50% yield) as the TFA salt. MS [M+H] + = 409.

[0292] Compound 183. 1-(4-(4-chlorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-3-(2-morpholinoethyl)urea Phenyl 4-(ethoxycarbonyl)-1,2,3-thiadiazol-5-ylcarbamate (100 mg, 0.34 mmol) and 2-morpholinoethanamine (66 mg, 0.51 mmol) were taken up in 1.5 mL of DMF. DIEA (118 μL, 0.68 mol) was added and the reaction was stirred at room temperature overnight. EtOAc was added and washed with saturated NaHCO3 and brine. The crude material was purified by normal phase chromatography (0-10% MeOH / DCM). 80 mg (72% yield) of a solid was produced.

[0293] Ethyl 5-(3-(2-morpholinoethyl)ureido)-1,2,3-thiadiazole-4-carboxylate (75 mg, 0.23 mmol) was taken up in 2 mL of MeOH / H2O (1 / 1) and lithium hydroxide (100 mg, 2.4 mmol) was added. The reaction was stirred at room temperature overnight. The volatiles were removed under vacuum and the solution was acidified with 4 mL of 1N HCl. The precipitate was filtered off to give 46 mg of 5-(3-(2-morpholinoethyl)ureido)-1,2,3-thiadiazole-4-carboxylic acid.

[0294] 5-(3-(2-morpholinoethyl)ureido)-1,2,3-thiadiazole-4-carboxylic acid (23 mg, 0.08 mmol), (4-chlorophenyl)methanamine (28 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give 1-(4-(4-chlorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-3-(2-morpholinoethyl)urea (28 mg, 65% yield) as the TFA salt. MS [M+H] + = 425.

[0295] Compound 184. 1-(4-(4-fluorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-3-(2-(pyridin-2-yl)ethyl)urea 5-(3-(2-(pyridin-2-yl)ethyl)ureido)-1,2,3-thiadiazole-4-carboxylic acid (30 mg, 0.1 mmol), (4-fluorophenyl)methanamine (25 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give 1-(4-(4-fluorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-3-(2-(pyridin-2-yl)ethyl)urea (48 mg, 93% yield) as the TFA salt. MS [M+H] + = 401.

[0296] Compound 185. 1-(4-(4-chlorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-3-(2-(pyridin-2-yl)ethyl)urea Phenyl 4-(ethoxycarbonyl)-1,2,3-thiadiazol-5-ylcarbamate (100 mg, 0.34 mmol) and 2-(pyridin-2-yl)ethanamine (62 mg, 0.51 mmol) were taken up in 1.5 mL of DMF. DIEA (118 μL, 0.68 mol) was added and the reaction was stirred at room temperature overnight. EtOAc was added and washed with saturated NaHCO3 and brine. The crude material was purified by normal phase chromatography (0-10% MeOH / DCM). 85 mg (78% yield) of a solid was produced.

[0297] Ethyl 5-(3-(2-(pyridin-2-yl)ethyl)ureido)-1,2,3-thiadiazole-4-carboxylate (77 mg, 0.24 mmol) was taken up in 2 mL of MeOH / H2O (1 / 1) and lithium hydroxide (100 mg, 2.4 mmol) was added. The reaction was stirred at room temperature overnight. The volatiles were removed in vacuo and the solution was acidified with 4 mL of 1N HCl. The precipitate was filtered off and 60 mg of the product was used as is.

[0298] 5-(3-(2-(pyridin-2-yl)ethyl)ureido)-1,2,3-thiadiazole-4-carboxylic acid (30 mg, 0.1 mmol), (4-chlorophenyl)methanamine (28 mg, 0.20 mmol), EDCI (38 mg, 0.20 mmol), and HOBt (30 mg, 0.20 mmol) were taken in 1.0 mL of DMF and stirred at room temperature overnight. The reaction was purified by preparative HPLC to give 1-(4-(4-chlorobenzylcarbamoyl)-1,2,3-thiadiazol-5-yl)-3-(2-(pyridin-2-yl)ethyl)urea (44 mg, 83% yield) as the TFA salt. MS [M+H] + = 417.

[0299] Compound 188. 5-Carbamoyl-4-methyl-2-(2-(p-tolyl)butanamido)thiophene-3-carboxylate methyl 2-(p-Tolyl)acetic acid (0.60 g, 4.0 mmol) was taken up in 16 mL of dry THF. The solution was cooled to -78 °C in a dry ice / acetone bath. n-Butylithium (3.4 mL, 0.0084 mol, 2.1 eq, 2.5 M in hexanes) was then added dropwise. The solution was stirred at 0 °C for 2 h. Iodoethane (0.40 mL, 4.8 mmol, 1.2 eq) was added slowly and the reaction was stirred at room temperature overnight. The reaction was quenched with water and the volatiles were removed under vacuum. 1N HCl was added to the solution and the product was extracted twice with diethyl ether. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM). 370 mg (52% yield) of 2-(p-tolyl)butanoic acid was produced as a white solid.

[0300] 2-(p-Tolyl)butanoic acid (370 mg, 2.07 mmol) was taken up in 3.7 mL of dry DCM under N2 atmosphere. Oxalyl chloride (196 μL, 2.28 mmol, 1.1 eq) was added to the solution along with one drop of dry DMF. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The acyl chloride was taken up in 4 mL of pyridine and methyl 2-amino-5-carbamoyl-4-methylthiophene-3-carboxylate (356 mg, 1.66 mmol) was added. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, followed by washing with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purification gave 196 mg (37% yield) of methyl 5-carbamoyl-4-methyl-2-(2-(p-tolyl)butanamido)thiophene-3-carboxylate as a solid TFA salt. MS [M+H] + = 375.

[0301] Compound 189. 5-Carbamoyl-2-(2-(2-fluorophenyl)butanamido)-4-methylthiophene-3-carboxylate methyl 2-(2-Fluorophenyl)acetic acid (0.62 g, 4.0 mmol) was taken up in 16 mL of dry THF. The solution was cooled to -78 °C in a dry ice / acetone bath. Then, n-butylithiuric acid (3.4 mL, 0.0084 mol, 2.1 eq, 2.5 M in hexanes) was added dropwise. The solution was stirred at 0 °C for 2 h. Iodoethane (0.40 mL, 4.8 mmol, 1.2 eq) was added slowly and the reaction was stirred at room temperature overnight. The reaction was quenched with water and the volatiles were removed under vacuum. 1N HCl was added to the solution and the product was extracted twice with diethyl ether. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM). 2-(2-Fluorophenyl)butanoic acid was obtained as a clear oil (260 mg, 36% yield).

[0302] 2-(2-Fluorophenyl)butanoic acid (260 mg, 1.42 mmol) was taken up in 2.6 mL of dry DCM under N2 atmosphere. Oxalyl chloride (134.7 μL, 1.570 mmol, 1.1 eq) was added to the solution along with one drop of dry DMF. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The acyl chloride was taken up in 4 mL of pyridine and methyl 2-amino-5-carbamoyl-4-methylthiophene-3-carboxylate (245 mg, 1.14 mmol) was added. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, followed by washing with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purification gave 203 mg (38% yield) of methyl 5-carbamoyl-2-(2-(2-fluorophenyl)butanamido)-4-methylthiophene-3-carboxylate as a solid TFA salt. MS [M+H] + = 379.

[0303] Compound 190. 5-Carbamoyl-4-methyl-2-(2-(4-(trifluoromethyl)phenyl)butanamido)thiophene-3-carboxylate methyl 2-(4-(trifluoromethyl)phenyl)acetic acid (0.50 g, 2.4 mmol) was taken up in 10 mL of dry toluene. The solution was cooled to -48 °C in a dry ice / acetonitrile bath. Lithium bis(trimethylsilyl)amide (0.86 g, 5.14 mmol, 2.1 eq) was then added dropwise. The solution was stirred at -48 °C for 1 h, iodoethane (0.23 mL, 2.9 mmol, 1.2 eq) was added slowly and the reaction was stirred at room temperature for 30 min. The reaction mixture was quenched with HCl (1 M) to pH = 1 and the product was extracted with EtOAc. The combined organic layers were dried (Na2SO4) and the solvent was removed under reduced pressure. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM). 296 mg (52% yield) of 2-(4-(trifluoromethyl)phenyl)butanoic acid was produced as a white solid.

[0304] 2-(4-(trifluoromethyl)phenyl)butanoic acid (80 mg, 0.34 mmol) was taken up in 1 mL of dry DCM under N2 atmosphere. Oxalyl chloride (32 μL, 0.37 mmol, 1.1 eq) was added to the solution along with one drop of dry DMF. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The acyl chloride was taken up in 2 mL of pyridine and methyl 2-amino-5-carbamoyl-4-methylthiophene-3-carboxylate (59 mg, 0.27 mmol) was added. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, followed by washing with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed under vacuum and purification gave 65 mg (44% yield) of methyl 5-carbamoyl-4-methyl-2-(2-(4-(trifluoromethyl)phenyl)butanamido)thiophene-3-carboxylate as a solid TFA salt. MS [M+H] + = 429.

[0305] Compound 191. 5-Carbamoyl-2-(2-(4-chlorophenyl)butanamido)-4-methylthiophene-3-carboxylate methyl 2-(4-Chlorophenyl)acetic acid (0.68 g, 4.0 mmol) was taken up in 16 mL of dry THF. The solution was cooled to -78 °C in a dry ice / acetone bath. Then, n-butylithiuric acid (3.4 mL, 0.0084 mol, 2.1 eq, 2.5 M in hexanes) was added dropwise. The solution was stirred at 0 °C for 2 h. Iodoethane (0.40 mL, 4.8 mmol, 1.2 eq) was added slowly and the reaction was stirred at room temperature overnight. The reaction was quenched with water and the volatiles were removed under vacuum. 1N HCl was added to the solution and the product was extracted twice with diethyl ether. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM). 450 mg (63% yield) of 2-(4-chlorophenyl)butanoic acid was produced as a clear oil.

[0306] 2-(4-Chlorophenyl)butanoic acid (450 mg, 2.52 mmol) was taken up in 5 mL of dry DCM under N2 atmosphere. Oxalyl chloride (238 μL, 2.77 mmol, 1.1 eq) was added to the solution along with one drop of dry DMF. The reaction was stirred at room temperature for 1 h. The solvent was removed under vacuum and the material was placed under high vacuum for 1 h. The acyl chloride was taken up in 6 mL of pyridine and methyl 2-amino-5-carbamoyl-4-methylthiophene-3-carboxylate (432 mg, 2.02 mmol) was added. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc was added, followed by washing with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed in vacuo and purification gave 465 mg (35% yield) of methyl 5-carbamoyl-2-(2-(4-chlorophenyl)butanamido)-4-methylthiophene-3-carboxylate as a solid TFA salt. MS [M+H] + = 394.87.

[0307] Compound 192. 5-(2-(benzo[d][1,3]dioxol-5-yl)butanamido)-3-methylthiophene-2,4-dicarboxylate 2-tert-butyl 4-methyl 2-(benzo[d][1,3]dioxol-6-yl)acetic acid (0.61 g, 3.37 mmol) was taken up in 15 mL of dry THF and the solution was cooled to -78 °C in a dry ice / acetone bath, then n-butyllithium in hexanes (3.0 mL, 7.0 mmol, 2.1 equiv, 2.5 M) was added dropwise. The solution was stirred at 0 °C for 2 h, then iodoethane (0.33 mL, 3.7 mmol, 1.1 equiv) was added slowly and the reaction was stirred at room temperature overnight. The reaction was quenched with 5 mL of water and the volatiles were removed in vacuo. To the resulting solution was added 1N HCl and the solution was extracted twice with diethyl ether. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM) to afford 2-(benzo[d][1,3]dioxol-6-yl)butanoic acid as a white solid (0.38 g, 54% yield).

[0308] 2-(Benzo[d][1,3]dioxol-6-yl)butanoic acid (0.235 g, 1.13 mmol) was taken up in 5 mL of dry DCM under N2 atmosphere. Oxalyl chloride (0.11 mL, 1.24 mmol, 1.1 equiv) was added to the solution along with 2 drops of dry DMF. The reaction was stirred at room temperature for 1 h. The progress of the reaction (i.e., formation of acyl chloride) was checked by quenching an aliquot of the reaction mixture with MeOH and was considered complete when only the methyl ester was observed by LC / MS analysis of the processed aliquot. The solvent was removed in vacuo and the material was placed under high vacuum for 1 h. 2-tert-butyl 4-methyl 5-amino-3-methylthiophene-2,4-dicarboxylate (0.28 g, 1.0 mmol) was added along with 5 mL of pyridine. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. 50 mL of EtOAc was added, followed by washing with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed in vacuo, and the crude material was purified by normal phase chromatography (0-20% EtOAc / DCM) to give the title compound as a white solid (0.18 g, 35% yield). MS: [M+H] + 462.

[0309] Compound 193. 2-(2-(benzo[d][1,3]dioxol-5-yl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate methyl 2-tert-Butyl 4-methyl 5-(2-(benzo[d][1,3]dioxol-5-yl)butanamido)-3-methylthiophene-2,4-dicarboxylate (60 mg, 0.13 mmol) was taken up in 0.5 mL of DCM and 0.25 mL of TFA and stirred for 1 h. The solvent was removed in vacuo and the material was placed under high vacuum for 1 h. DMF (1 mL) was then added along with ammonium chloride (65 mg, 1.02 mmol), EDCI (37 mg, 0.2 mmol), HOBt (30 mg, 0.2 mmol) and triethylamine (0.17 mL, 1.2 mmol). The reaction was stirred at room temperature overnight. The reaction was quenched with water and extracted with EtOAc. The crude material was purified by preparative HPLC to give the title compound as a white powder (30 mg, 57% yield). MS: [M+H] + 405.

[0310] Compound 194. 5-(2-(3,5-difluorophenyl)butanamido)-3-methylthiophene-2,4-dicarboxylate 2-tert-butyl 4-methyl 2-(3,5-Difluorophenyl)acetic acid (0.655 g, 3.8 mmol) was taken up in 15 mL of dry THF and the solution was cooled to -78 °C via a dry ice / acetone bath, then n-butyllithium in hexanes (3.4 mL, 8.0 mmol, 2.1 equiv, 2.5 M) was added dropwise. The solution was stirred at 0 °C for 2 h, then iodoethane (0.37 mL, 4.2 mmol, 1.1 equiv) was added slowly and the reaction was stirred at room temperature overnight. The reaction was quenched with 5 mL of water and the volatiles were removed in vacuo. To the resulting solution was added 1N HCl and the solution was extracted twice with diethyl ether. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM) to give 2-(3,5-difluorophenyl)butanoic acid as a white solid (0.42 g, 55% yield).

[0311] 2-(3,5-Difluorophenyl)butanoic acid (0.42 g, 2.1 mmol) was taken up in 10 mL of dry DCM under N2 atmosphere. Oxalyl chloride (0.20 mL, 2.3 mmol, 1.1 equiv) was added to the solution along with 2 drops of dry DMF. The reaction was stirred at room temperature for 1 h. The progress of the reaction (i.e., formation of acyl chloride) was checked by quenching an aliquot of the reaction mixture with MeOH and was considered complete when only the methyl ester was observed by LC / MS analysis of the processed aliquot. The solvent was removed in vacuo and the material was placed under high vacuum for 1 h. 2-tert-butyl 4-methyl 5-amino-3-methylthiophene-2,4-dicarboxylate (0.51 g, 1.9 mmol) was added along with 5 mL of pyridine. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc (150 mL) was added, followed by washing with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed in vacuo and the crude material was purified by normal phase chromatography (0-20% EtOAc / DCM) to give the title compound as a white solid (0.49 g, 52% yield). MS: [M+H] + 454.

[0312] Compound 195. 2-(2-(3,5-difluorophenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate methyl 2-tert-Butyl 4-methyl 5-(2-(3,5-difluorophenyl)butanamido)-3-methylthiophene-2,4-dicarboxylate (60 mg, 0.13 mmol) was taken up in 1.0 mL of DCM and 0.50 mL of TFA and stirred for 1 h. The solvent was removed in vacuo and the material was placed under high vacuum for 1 h. DMF (2 mL) was added along with ammonium chloride (65 mg, 1.02 mmol), EDCI (37 mg, 0.2 mmol), HOBt (30 mg, 0.2 mmol) and triethylamine (0.17 mL, 1.2 mmol). The reaction was stirred at room temperature overnight. The reaction was quenched with water and extracted with EtOAc. The crude material was purified by preparative HPLC to give the title compound as a white powder (10 mg, 20% yield). MS: [M+H] + 397.

[0313] Compound 196. 5-(2-(4-cyanophenyl)butanamido)-3-methylthiophene-2,4-dicarboxylate 2-tert-butyl 4-methyl 2-(4-Cyanophenyl)acetic acid (0.745 g, 4.6 mmol) was taken up in 20 mL of dry THF and the solution was cooled to -78 °C via a dry ice / acetone bath, then n-butyllithium in hexanes (4.0 mL, 9.7 mmol, 2.1 equiv, 2.5 M) was added dropwise. The solution was stirred at 0 °C for 2 h, then iodoethane (0.45 mL, 5.5 mmol, 1.1 equiv) was added slowly and the reaction was stirred at room temperature overnight. The reaction was quenched with 5 mL of water and the volatiles were removed in vacuo. To the resulting solution was added 1N HCl and the solution was extracted twice with diethyl ether. The crude material was purified using normal phase chromatography (0-5% MeOH / DCM) to afford 2-(4-cyanophenyl)butanoic acid as a white solid (0.41 g, 47% yield).

[0314] 2-(4-Cyanophenyl)butanoic acid (0.41 g, 2.1 mmol) was taken up in 10 mL of dry DCM under N2 atmosphere. Oxalyl chloride (0.20 mL, 2.3 mmol, 1.1 equiv.) was added to the solution along with 2 drops of dry DMF. The progress of the reaction (i.e., formation of acyl chloride) was checked by quenching an aliquot of the reaction mixture with MeOH and was considered complete when only the methyl ester was observed by LC / MS analysis of the processed aliquot. The solvent was removed in vacuo and the material was placed under high vacuum for 1 h. 2-tert-butyl 4-methyl 5-amino-3-methylthiophene-2,4-dicarboxylate (0.51 g, 1.9 mmol) was added along with 5 mL of pyridine. A catalytic amount of DMAP was added and the reaction was stirred at room temperature overnight. EtOAc (150 mL) was added, followed by washing with 1N HCl, saturated sodium bicarbonate, and brine. The solvent was removed in vacuo and purified by normal phase chromatography (0-20% EtOAc / DCM) to give the title compound as a white solid (0.35 g, 38% yield). MS: [M+H] + 443.

[0315] Compound 197. Methyl 2-(2-(4-cyanophenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxylate 2-tert-Butyl 4-methyl 5-(2-(4-cyanophenyl)butanamido)-3-methylthiophene-2,4-dicarboxylate (66 mg, 0.15 mmol) was taken up in 1.0 mL of DCM and 0.50 mL of TFA and stirred for 1 h. The solvent was removed in vacuo and the material was placed under high vacuum for 1 h. 2 mL of DMF was added along with ammonium chloride (65 mg, 1.2 mmol), EDCI (37 mg, 0.2 mmol), HOBt (30 mg, 0.2 mmol) and triethylamine (0.17 mL, 1.2 mmol). The reaction was stirred at room temperature overnight. The reaction was quenched with water and extracted with EtOAc. The crude material was purified by preparative HPLC to give the title compound as a white powder (42 mg, 73% yield). MS: [M+H] + 386.

[0316] Compound 198. 5-(2-(4-(trifluoromethyl)phenyl)butanamido)-3-methylthiophene-2,4-dicarboxylate 2-tert-butyl 4-methyl 2-(4-(trifluoromethyl)phenyl)butanoic acid (0.965 g, 4.16 mmol) was taken up in 50 mL of anhydrous DCM under nitrogen atmosphere. Oxalyl chloride (2.0 M in DCM, 2.08 mL, 4.16 mmol) was added to the solution along with 2 drops of anhydrous DMF. The reaction was stirred at room temperature for 1 h. The solvent was evaporated and 2-tert-butyl 4-methyl-5-amino-3-methylthiophene-2,4-dicarboxylate (0.51 g, 1.9 mmol) was added along with 10 mL of pyridine. 4-Dimethylaminopyridine (51 mg, 0.42 mmol) was added and the reaction was stirred at room temperature overnight. The mixture was diluted with brine and extracted with ethyl acetate. The extract was washed with 1N HCl and then concentrated. The resulting solid was chromatographed (40 g silica column, EtOAc / hexanes) to give the title compound as an off-white solid (0.61 g, 66% yield). MS: [M+H] + 486.2.

[0317] Compound 199. 4-(Methoxycarbonyl)-5-(2-(4-(trifluoromethyl)phenyl)butanamido)-3-methylthiophene-2-carboxylic acid 2-tert-Butyl 4-methyl 5-(2-(4-(trifluoromethyl)phenyl)butanamido)-3-methylthiophene-2,4-dicarboxylate (0.60 g, 1.25 mmol) was dissolved in DCM (18 mL) and then trifluoroacetic acid (2 mL) was added. The solution was stirred at room temperature overnight and then concentrated to give the title compound as an off-white solid (0.53 g, quantitative yield). MS: [M+H] + 430.1.

[0318] Compound 200. 5-(2-aminoethylcarbamoyl)-2-(2-(4-(trifluoromethyl)phenyl)butanamido)-4-methylthiophene-3-carboxylate methyl 4-(Methoxycarbonyl)-5-(2-(4-(trifluoromethyl)phenyl)butanamido)-3-methylthiophene-2-carboxylic acid (0.384 g, 0.90 mmol) was dissolved in anhydrous dichloromethane (10 mL). Oxalyl chloride (2M in dichloromethane, 0.450 mL) and dimethylformamide (approximately 5 drops) were added. The reaction was stirred at room temperature for 1 hour. Then, tert-butyl 2-aminoethylcarbamate (0.173 g, 1.08 mmol) and diisopropylethylamine (0.289 g, 2.24 mmol) were added and the reaction was stirred at room temperature overnight. The reaction was diluted with saturated aqueous sodium bicarbonate and extracted with dichloromethane. The extract was concentrated and subjected to chromatography (12 g silica column, hexane / ethyl acetate). The resulting product was dissolved in a solution of HCl / dioxane (4N, 20 mL). After 2 hours, the solution was lyophilized to give the hydrochloride salt of the title compound as an off-white powder (0.305 g, 67% yield). MS: [M+H] + 472.4

[0319] Compound 201. 2-[2-(4-fluorophenyl)butanamido]-4-methyl-5-(piperazine-1-carbonyl)thiophene-3-carboxylate methyl 4-(Methoxycarbonyl)-5-(2-(4-fluorophenyl)butanamido)-3-methylthiophene-2-carboxylic acid (0.18 g, 0.47 mmol) was dissolved in anhydrous dichloromethane (10 mL). Oxalyl chloride (2M in dichloromethane, 0.237 mL) and dimethylformamide (approximately 5 drops) were added. The reaction was stirred at room temperature for 1 hour. Piperazine (0.122 g, 1.41 mmol) was added and the reaction was stirred at room temperature overnight. The reaction was diluted with brine and extracted with ethyl acetate. The extract was diluted with saturated aqueous sodium bicarbonate and extracted with dichloromethane. The extract was concentrated and chromatographed (12 g silica column, hexane / ethyl acetate). The resulting product was dissolved in a solution of HCl / dioxane (4N, 3 mL) and lyophilized to give the hydrochloride salt of the title compound as an off-white powder (0.10 g, 44% yield). MS: [M+H] + 448.2.

[0320] Compound 202. Methyl 5-((3-((tert-butoxycarbonyl)amino)propyl)carbamoyl)-2-(2-(4-fluorophenyl)butanamido)-4-methylthiophene-3-carboxylate 4-(Methoxycarbonyl)-5-(2-(4-fluorophenyl)butanamido)-3-methylthiophene-2-carboxylic acid (0.090 g, 0.24 mmol) was dissolved in anhydrous dimethylformamide (2 mL) and then PyBOP (0.146 g, 0.28 mmol) was added. The reaction was stirred at room temperature for 15 minutes. Diisopropylethylamine (0.077 g, 0.60 mmol) and tert-butyl 3-aminopropylcarbamate (0.050 g, 0.28 mmol) were then added and the reaction was stirred at room temperature overnight. The reaction was diluted with brine and extracted with ethyl acetate. The extract was concentrated and chromatographed (12 g silica column; hexane / ethyl acetate) to give the title compound as a white solid (0.085 g, 66% yield). MS: [M+H] + 536.5.

[0321] Compound 203. Methyl 5-(3-aminopropylcarbamoyl)-2-(2-(4-fluorophenyl)butanamido)-4-methylthiophene-3-carboxylate tert-Butyl 3-(methyl 2-(2-(4-fluorophenyl)butanamido)-5-carbamoyl-4-methylthiophene-3-carboxyloyl)propylcarbamate (0.080 g, 0.15 mmol) was dissolved in a solution of hydrogen chloride in dioxane (4N, 2 mL). After 2 h, the solution was lyophilized to give the hydrochloride salt of the title compound as a white powder (0.073 g, quantitative yield). MS: [M+H] + 436.5.

[0322] Example 5: Biological Assays I C 50 In vitro DNA synthesis assay: Continuous DNA synthesis assay Rapid plate assays were performed as previously described (Lin & Ricciardi, 2000, J. Virol. Methods 88:219-225). Briefly, 5' biotinylated 100 nucleotide templates containing an adenine only at their 5' distal end were annealed to a 15 nucleotide primer at their 3' end and attached to streptavidin-coated 96-plate wells (Roche Applied Science). DNA synthesis was carried out in 50 μL reaction mixtures containing 100 mM (NH)2SO4, 20 mM Tris-HCl (pH 7.5), 3 mM MgCl2, 0.1 mM EDTA, 0.5 mM DTT, 2% glycerol, 40 μg / ml BSA, 5 μM dATP, 5 μM dCTP, 5 μM dGTP, 1 μM digoxigenin-11-dUTP, and E9 / A20 / D4 proteins. Vaccinia virus vD4 or molluscum contaminant mD4 and TNT reticulocyte lysate containing vEC50 and vA20 or in vitro translated luciferase were used as negative controls and were added to the reaction mixtures. After incubation at 37°C for 30 min, the plates were washed extensively with phosphate-buffered saline (PBS). The wells were then incubated with anti-digoxigenin peroxidase antibody (Roche) for 1 h at 37°C and then washed with PBS. The substrate 2,2'-azino-bis(3-ethylbenzthiazoline)-sulfonate (Roche) was added and the plate was developed by gently rocking. DNA synthesis was quantified by measuring the absorbance of each reaction at 405 nm with a microplate reader (Tecan). Experiments were performed in triplicate and independently repeated at least twice. Increasing concentrations of compounds of interest were added independently to all reaction mixtures to determine IC 50 Got the value.

[0323] EC 50 Plaque Reduction Assay: Reduction of viral plaques Viral plaque reduction assays were performed in triplicate using BSC-1 cells as previously described (Nuth, et al., 2011, J. Med. Chem. 54:3260-3267) and repeated independently with compounds of interest. Cells were adsorbed and infected with poxvirus at 80 PFU / well in 100 μL of growth medium in 48-well plates for 1 h, followed by treatment with compounds for 16 h. Cells were stained, plaques were counted, and data were plotted in GraphPad Prism.

[0324] Table 1. Summary of DSF experiments with 25 and 50 μM compound treatment. For TIFF2024536861000101.tif41128D4, m =42.24±0.66° C. Values ​​for compound 10 reflect n=9 and n=3 for cidofovir (CDV) and tecovirimat (ST-246).

[0325] Table 2. Compounds and biological activities TIFF2024536861000102.tif46164TIFF2024536861000103.tif224164TIFF2024536861000104.tif246164T IFF2024536861000105.tif243164TIFF2024536861000106.tif255164TIFF2024536861000107.tif202164TI FF2024536861000108.tif235164TIFF2024536861000109.tif224164TIFF2024536861000110.tif231164TIF F2024536861000111.tif241164TIFF2024536861000112.tif255164TIFF2024536861000113.tif214164TIFF 2024536861000114.tif255162TIFF2024536861000115.tif212164TIFF2024536861000116.tif253164TIFF2 024536861000117.tif225164TIFF2024536861000118.tif235164TIFF2024536861000119.tif246164TIFF20 24536861000120.tif190164TIFF2024536861000121.tif248164TIFF2024536861000122.tif236164TIFF202 4536861000123.tif243164TIFF2024536861000124.tif245164TIFF2024536861000125.tif52164 Compound 199(CC 50 =>100μM; ATP assay)

[0326] Example 6: In vitro skin permeation studies A non-GLP in vitro skin permeation study was conducted with compounds 99 and 111. The project included preformulation studies producing five prototype formulations for each compound, development of a robust method to measure percutaneous permeation, and skin permeation studies using human cadaver skin in a vertical diffusion (Franz) cell model. The diffusion chamber was designed to maintain skin slices at a temperature and humidity representative of typical in vivo conditions. The vertical diffusion cell human skin dosing model has historical precedent for accurately predicting in vivo percutaneous absorption kinetics.

[0327] Table 3: Formulation compounds TIFF2024536861000126.tif115160 a Determined by NMR and LC / MS; b pKa calculated for the HCl salt of the indicated compound.

[0328] Compound solubility for compounds 99 and 111 was assessed visually and by HPLC in 42 excipients. Based on these results, 10 theoretical solvent mixtures were designed for each compound, and of these 10 mixtures, 3 mixtures were prepared and tested for solubility at a target final concentration of 10 mg / mL (1%). Samples were prepared by dissolving 1.4% of API in the mixture, stirring overnight, filtering, and then analyzing the solution by HPLC for % recovery and target concentration.

[0329] Table 4A. Formulations of Compound 111 TIFF2024536861000127.tif135160 a Values ​​are given as %w / w

[0330] Table 4B: Formulations of Compound 99 TIFF2024536861000128.tif135160 a Values ​​are given as %w / w

[0331] For compound 111, all three blends were able to achieve the 1% target concentration, and for 99, two blends were able to reach the 1% target concentration. From these results, five prototype formulations were designed for each compound, including gels and ointments with different levels of permeation enhancers (Tables 5A-5B). Optimized blending protocols were developed to reach the 1% target concentration in all five formulations for each compound. For compound 111, F1 and F2 are ointments, and F3, F4, and F5 are gels. For compound 99, F8 is an ointment, and F6, F7, F9, and F10 are gels. For both compounds 99 and 111, pilot 2-week stability studies were conducted at room temperature with the three formulations. The percent recoveries for the T=0 to T=2 week samples showed API loss of less than 2.0% for all formulations, representing good product stability.

[0332] Five formulations of each of compounds 99 and 111 were tested in triplicate in an in vitro skin permeation (IVPT) study using a Franz cell apparatus as described elsewhere herein to measure compound levels in the stratum corneum, epidermis, dermis and receptor medium using human dermatome skin. For this study, human torso skin was used rather than a membrane. First, a method was developed to identify the receptor medium in the Franz cell apparatus reservoir with the highest compound solubility, so that complete penetration was not underestimated due to low solubility. A total of six receptor medium buffers were evaluated, and 1x PBS / 4% BSA was found to provide the highest solubility.

[0333] Furthermore, the solubility values ​​are sufficient to detect up to 143% and 90% of the applied doses of 99 and 111, respectively, in a typical IVPT study, which far exceeds the amount of compound expected to fully penetrate the skin.

[0334] The total percent of compound recovered in all four samples for each formulation is shown (Figures 17A-17B). For formulations F1-F4, less than 5% of 111 was recovered in four samples, but it is assumed that most remained in the formulation at the skin surface and was washed off. F5 gel showed the greatest skin penetration, with the highest levels of compound 111 found in the stratum corneum. Although lower than the stratum corneum, F5 also recovered significant levels of 111 in the epidermis and dermis (5.1% of total applied amount).

[0335] Stratum corneum levels of 99 were highest in F9 and F10, whereas epidermal and dermal levels were more comparable in all five formulations, with the highest levels obtained in F7 (6.1% of total applied dose) and F9 (5.1% of total applied dose). Negligible amounts were detected in the receptor medium in all formulations for each compound.

[0336] For 111, only two formulations showed complete skin penetration (recovered in the receptor medium), with the highest value being obtained for F5 (4.2 ng, i.e. 0.004% of the total applied dose). For 99, all five formulations showed complete skin penetration, with the highest value being obtained for F9 (95 ng, i.e. 0.09% of the total applied dose). Interestingly, F7 showed the highest penetration into the epidermis and dermis, but the least complete penetration, with only 18.9 ng recovered in the receptor medium (0.018% of the total applied dose). Only very low levels of the 99 acid analogue (99b) were detected in the epidermis for all five formulations (3.4–10 ng in total), representing a low degree of metabolic transformation in this study.

[0337] Because molluscum contagiosum grows only in the epidermis of human skin, the goal is to deliver effective antiviral levels of compound to the epidermis while minimizing systemic exposure. To better evaluate the significance of the 99 and 111 compound levels obtained with the more skin-penetrating formulations, weights (i.e., ng / mg tissue sample) were converted to μM levels, assuming 1 g of tissue equals 1 mL. While it is recognized that there are many variables in predicting antiviral concentrations of compounds in tissues, including the presence of multiple tissue compartments (e.g., cells, cell types, interstitial fluid, matrix, different protein and lipid concentrations) and their effects on compound distribution and activity, this is believed to be a useful first step in assessing whether there is a realistic chance of achieving effective compound concentrations in target tissues following local delivery.

[0338] Table 5A: Skin levels of compound (compound 111) TIFF2024536861000129.tif71160

[0339] Table 5B: Skin levels of compound (compound 99) TIFF2024536861000130.tif162160

[0340] The mean ng / mL and calculated mean μM concentrations of compound in the epidermis and dermis obtained for formulations F1 and F5 containing 111 and formulations F6-F10 containing 99 are provided herein (Tables 6A-6B). These data are also presented graphically in Figures 18A-18B, where the mean μM concentration of compound is plotted for each formulation.

[0341] The antiviral activity of compounds 111 and 99 in a cell plaque assay against hybrid mD4 / vaccinia hybrid virus was EC 50 The epidermal concentrations of 111 in F5 and 99 in all five formulations were found to be EC 50 Concentrations exceed those of the 100-kDa 2 ...

[0342] Thus, in vitro skin permeation studies confirmed that formulations of compounds 111 and 99 appear to deliver effective concentrations of the antiviral compounds to the epidermis of human cadaver skin following topical administration. Complete penetration through all skin layers was low, consistent with low systemic exposure, and negligible metabolic conversion of 99 to its acid analogue.

[0343] Stability studies were performed on 99 formulated in F7 gel (8 mg / mL) at RT and 40° C. 99 was obtained with 98% recovery over the 3-month study period, indicating excellent stability.

[0344] Additionally, acute skin irritation studies were conducted in rabbits with 10 mg / mL of 99 in the F7 gel formulation. Briefly, the gel was applied to a shaved skin area, covered for 24 hours, and irritation was measured daily for 3 days after removal of the cover using the Draize scoring system. The F7 gel formulation containing 99 was scored as non-irritating. Similarly, 99 formulated in a gel was scored as non-irritating on human cadaver skin.

[0345] Sequence Listing TIFF2024536861000131.tif23158TIFF2024536861000132.tif23158

[0346] Numbering aspects The following illustrative aspects are provided, the numbering of which is not to be construed as indicating order of importance.

[0347] Embodiment 1 provides a compound of formula (I), or a salt, solvate, enantiomer, diastereoisomer, geometric isomer, or tautomer thereof: TIFF2024536861000133.tif23128 formula, X is CR 1 or N; Y is CR 2 or N; R 1is H, optionally substituted C1-C6 alkyl, -C(=O)NR 6 R 6 , -C(=O)OR 6 , or R 8 and; R 2 is H or optionally substituted C1-C6 alkyl; R 3 H, -CN, -C(=O)OR 6 , -C(=O)NR 6 R 6 , optionally substituted phenyl, or optionally substituted C1-C6 alkyl; R 4 -C(=O)OR 6 or R 8 and; R 5 is, at each occurrence, independently, optionally substituted C1-C6 alkyl or optionally substituted phenyl; R 6 is independently at each occurrence H or an optionally substituted C1-C6 alkyl; or two R's 6 together with the N atom to which they are both attached form an optionally substituted 4- to 7-membered heterocyclyl; R 8 is -C(=O)NH(optionally substituted acyl), -N(optionally substituted acyl)C(=O)R 7 , -NR 6 C(=O)R 7 or -NR 6 C(=O)NR 6 R 7 and; R 7 each occurrence independently represents optionally substituted C1-C6 alkyl, optionally substituted cycloalkyl, CH(optionally substituted heterocyclyl)(R 5 ), CH(R 5 )(R 5 ), or an optionally substituted 4- to 7-membered heterocyclyl; or R 6 and R 7together with the N atom to which they are both attached form an optionally substituted 4- to 7-membered heterocyclyl; However, (I) is R 8 contains one; and With the proviso that the compound is not selected from the group consisting of compounds 1, 2, 4, 5, 8, 9, 15, 16, 17, 19, 24, 26, 29, 31, 40, 66, 72, 76, 77, 78, 83, 84, and 86.

[0348] In the second aspect, X is CR 1 and Y is CR 2 The compound of embodiment 1 is provided,

[0349] Embodiment 3 provides a compound of embodiment 1, wherein X is N and Y is N.

[0350] In the fourth aspect, X is CR 1 and Y is N.

[0351] Aspect 5 is where X is N and Y is CR 2 The compound of embodiment 1 is provided,

[0352] Aspect 6 is The compound of any one of embodiments 1 to 5 is provided, wherein the compound is selected from the group consisting of: TIFF2024536861000134.tif26131.

[0353] Aspect 7 is R 4 R 8 The compound of any one of embodiments 1-6 is provided,

[0354] Embodiment 8 provides a compound of any one of embodiments 1-7, wherein at least one of the following is true: R 8 -NR 6 C(=O)R 7 and R 6 is H and R 7 is CH(CH2CH3)Ph; R 8 -NR 6C(=O)R 7 and R 6 is H and R 7 is CH(CH2CH3)(4-F-Ph); R 8 -NR 6 C(=O)NR 6 R 7 and R 6 is H and NR 6 R 7 is 4-phenyl-piperazin-1-yl; and R 8 -NR 6 C(=O)NR 6 R 7 and R 6 is H and NR 6 R 7 is 4-(2-pyridyl)-piperazin-1-yl.

[0355] Aspect 9 provides a compound of any one of aspects 1-8, wherein at least one of the following is true: R 3 -C(=O)OR 6 and R 6 is CH3; R 3 -C(=O)OR 6 and R 6 is CH2CH3; and R 3 is -C(=O)NR 6 R 6 and NR 6 R 6 is NH(CH2-aryl), wherein the aryl is selected from the group consisting of phenyl, 4-fluorophenyl, 4-chlorophenyl, and 4-trifluoromethylphenyl.

[0356] Aspect 10 is a compound comprising R 1 and R 4 One of The compound of any one of embodiments 1 to 6 is provided, wherein the compound is selected from the group consisting of: During the ceremony, Ra1 and R a2 each, if present, is independently selected from the group consisting of H and optionally substituted C1-C6 alkyl; R b1 , R b2 , R b3 , R b4 , and R b5 are each independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 haloalkyl, halogen, CN, and NO2, if present; R b1 , R b2 , R b3 , R b4 , and R b5 Two vicinal substituents selected from the group consisting of, together with the atoms to which they are attached, optionally substituted C2-C 10 may form a heterocycloalkyl; A 1 C2 to C 10 Heteroaryl and optionally substituted C2-C 10 heterocycloalkyl; G 1 is a bond and C(R c6 )(R c7 ) selected from the group consisting of; G 2 is a bond and C(R c8 )(R c9 ) selected from the group consisting of; G 3 is a bond and C(R c10 )(R c11 ) selected from the group consisting of; R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11are each independently selected from the group consisting of H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxy, optionally substituted C1-C6 haloalkyl, and halogen, if present; R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 Two vicinal substituents selected from the group consisting of, together with the atoms to which they are attached, optionally substituted C6-C 10 may form an aryl; Z 1 is N and CR c9 and Z 2 is N and CR b5 is selected from the group consisting of:

[0357] Embodiment 11 provides a compound of embodiment 10, wherein one of the following is true: (a)R a1 is H and R a2 is ethyl; or (b)R a1 is ethyl, and R a2 is H.

[0358] Aspect 12 is R b1 , R b2 , R b3 , R b4 , and R b5 12. The compound of embodiment 10 or 11, wherein each of, if present, is independently selected from the group consisting of H, Me, OMe, F, Cl, CF3, CN, and NO2.

[0359] Aspect 13 is R c1 , R c2 , R c3 , R c4 , R c5 , Rc6 , R c7 , R c8 , R c9 , R c10 , and R c11 Each of, when present, is independently selected from the group consisting of H and Ph.

[0360] Embodiment 14 provides a compound of embodiment 10 or 13, wherein at least one of the following is true: (a)G 1 , G 2 , and G 3 None of the are bonds; (b) G 1 , G 2 , and G 3 One of them is a bond; (c)G 1 , G 2 , and G 3 two of which are bonds; and (d) G 1 , G 2 , and G 3 Each of is a bond.

[0361] Aspect 15 is A 1 but The compound of embodiment 10 is provided, wherein the compound is selected from the group consisting of: TIFF2024536861000136.tif13128.

[0362] Embodiment 16 provides a compound of any one of embodiments 10-15, wherein at least one of the following is true: (a) R 2 is an optionally substituted C1-C6 alkyl, R 3 is C(=O)OR 6 and R 1 and R 4 One of TIFF2024536861000137.tif24128, and R b1 , R b2, R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF3, CN, and NO2 is selected from the group consisting of R b1 , R b2 , R b3 , R b4 , and R b5 two vicinal substituents selected from the group consisting of taken together to form methylenedioxy; (b) R 3 is an optionally substituted C1-C6 alkyl, R 2 is C(=O)OR 6 and R 1 and R 4 One of TIFF2024536861000138.tif24128, and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF3, CN, and NO2 is selected from the group consisting of R b1 , R b2 , R b3 , R b4 , and R b5 two vicinal substituents selected from the group consisting of taken together to form methylenedioxy; (c) R 2 is an optionally substituted C1-C6 alkyl, R 3 is CN, R 1 and R 4 One of TIFF2024536861000139.tif24128, and R b1 , R b2 , R b3 , R b4 , and Rb5 At least one selected from the group consisting of Me, OMe, F, Cl, CF3, CN, and NO2 is selected from the group consisting of R b1 , R b2 , R b3 , R b4 , and R b5 two vicinal substituents selected from the group consisting of taken together to form methylenedioxy; (d) R 2 is C(=O)OR 6 and R 1 and R 4 One of TIFF2024536861000140.tif24128, and R 3 and R 1 or R 4 Of these, not more than one is a C1-C6 alkyl; (e) R 1 is H, R 2 is H, R 3 is -C(=O)NR 6 R 6 and R 6 At most one occurrence of is H; (f) R 1 teeth TIFF2024536861000141.tif24128, R 2 is methyl, R 3 is H, and R 4 is selected from the group consisting of C(=O)O(C1 alkyl), C(=O)O(optionally substituted C3 alkyl), C(=O)O(optionally substituted C4 alkyl), C(=O)O(optionally substituted C5 alkyl), and C(=O)O(optionally substituted C6 alkyl); (g) R 4teeth TIFF2024536861000142.tif24128, R 3 is methyl, R 2 is H, and R 1 is selected from the group consisting of C(=O)O(C1 alkyl), C(=O)O(optionally substituted C3 alkyl), C(=O)O(optionally substituted C4 alkyl), C(=O)O(optionally substituted C5 alkyl), and C(=O)O(optionally substituted C6 alkyl); (h) R 1 is C(=O)NH2, R 4 teeth TIFF2024536861000143.tif24128, G 1 , G 2 , and G 3 At most one of is a bond, and R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 A pair of vicinal substituents selected from the group consisting of, together with the atom to which they are attached, is C6-C 10 Forming aryl; (i) R 1 is C(=O)NH2, R 4 teeth TIFF2024536861000144.tif24128, G 1 , G 2 , and G 3 At most one of is a bond, and R c2 , R c3 , R c4 , R c5, R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 A pair of vicinal substituents selected from the group consisting of, together with the atom to which they are attached, is C6-C 10 Forming aryl; (j) Y is N, X is CR 1 and R 1 and R 4 One of TIFF2024536861000145.tif24128, and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF3, CN, and NO2 is selected from the group consisting of R b1 , R b2 , R b3 , R b4 , and R b5 two vicinal substituents selected from the group consisting of taken together to form methylenedioxy; (k) X is N, Y is CR 2 and R 4 teeth TIFF2024536861000146.tif24128, and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF3, CN, and NO2 is selected from the group consisting of R b1 , R b2 , R b3 , R b4 , and R b5two vicinal substituents selected from the group consisting of taken together to form methylenedioxy; (l) X is N, Y is N, R 4 teeth TIFF2024536861000147.tif31128, and Z 1 is CR c9 and (m) X is N, Y is N, R 4 teeth TIFF2024536861000148.tif31128, Z 2 is CR b5 and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of is selected from the group consisting of halogen, C1-C6 alkyl, NO2, and CN.

[0363] Aspect 17 is R 1 is H, Me, C(=O)NH2, C(=O)NMe2, C(=O)NEt2, C(=O)OEt, C(=O)OMe, C(=O)OEt, C(=O)Ot-Bu, C(=O)O(CH2)2NH2, C(=O)NH(CH2)3NH2, The compound of any one of embodiments 1-16 is provided, wherein the compound is selected from the group consisting of: TIFF2024536861000149.tif198159TIFF2024536861000150.tif57128.

[0364] Aspect 18 is R 4 is Me, C(=O)NH2, C(=O)NMe2, C(=O)NEt2, C(=O)OEt, C(=O)OMe, C(=O)OEt, C(=O)Ot-Bu, C(=O)O(CH2)2NH2, C(=O)NH(CH2)3NH2, The compound of any one of embodiments 1-17 is provided, wherein the compound is selected from the group consisting of: TIFF2024536861000151.tif152148TIFF2024536861000152.tif103159.

[0365] Aspect 19 is R 2 is selected from the group consisting of H, Me and Et.

[0366] Aspect 20 is R 3 H, Me, Et, C(=O)OMe, C(=O)OEt, C(=O)NH2, CN, Ph, 4-trifluoromethylphenyl, 4-fluorophenyl, The compound of any one of embodiments 1-19 is provided, wherein the compound is selected from the group consisting of: TIFF2024536861000153.tif68140.

[0367] Aspect 21 is a compound of Compounds 22, 32, 33, 34, 35, 36, 39, 47, 50, 57, 68, 75, 82, 87, 89, 90, 95, 96, 97, 98, 99, 106, 109, 110, 111, 112, 114, 115, 116, 118, 119, 123, 124, 127, 130, 131, 132, 134, 135, 144, 153, 1 54, 155, 159, 160, 161, 162, 163, 165, 167, 168, 169, 170, 172, 173, 174, 175, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, and 191.

[0368] Aspect 22 is where alkyl or cycloalkyl, at each occurrence, is independently selected from C1-C6 alkyl, halo, carbonyl (C=O), -OR, optionally substituted phenyl, optionally substituted heteroaryl, optionally substituted heterocyclyl, -N(R)(R), -N(R)-(C=O)R, -C(=O)R, -C(=O)(optionally substituted phenyl), -C(=O)(optionally substituted heteroaryl), -C(=O)N(R)(R), -C(=O)(CH2) 0~3 and -OR, -S(=O)2R, and -SON(R)(R), where R, at each occurrence, is independently selected from the group consisting of H, C1-C6 alkyl, and C3-C8 cycloalkyl.

[0369] Embodiment 23 provides a compound of any one of embodiments 1-22, wherein phenyl, at each occurrence, is optionally substituted with at least one substituent selected from the group consisting of C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 haloalkoxy, halo, -CN, -OR, -N(R)(R), -NO2, -S(=O)2N(R)(R), acyl, and C1-C6 alkoxycarbonyl, and R, at each occurrence, is independently selected from the group consisting of H, C1-C6 alkyl, and C3-C8 cycloalkyl.

[0370] Embodiment 24 provides a compound of any one of embodiments 1-23, wherein phenyl, at each occurrence, is optionally substituted with at least one substituent independently selected from the group consisting of C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 haloalkoxy, halo, -CN, -OR, -N(R)(R), and C1-C6 alkoxycarbonyl, and R, at each occurrence, is independently selected from the group consisting of H, C1-C6 alkyl, and C3-C8 cycloalkyl.

[0371] Embodiment 25 provides a pharmaceutical composition comprising at least one compound of any one of embodiments 1-24 and at least one pharma- ceutically acceptable excipient.

[0372] Embodiment 26 provides a pharmaceutical composition comprising at least one compound selected from the group consisting of compounds 1, 2, 4, 5, 8, 9, 15, 16, 17, 19, 24, 26, 29, 31, 40, 66, 72, 76, 77, 78, 83, 84, 86, 99, and 111, and at least one pharma- ceutically acceptable excipient.

[0373] Embodiment 27 provides the pharmaceutical composition of embodiment 25 or 26, wherein at least one compound is compound 111.

[0374] Aspect 28 is a pharmaceutical composition comprising the at least one pharma- ceutically acceptable excipient selected from the group consisting of water, polyethylene glycol (PEG) 400, PEG 300, propylene glycol (PG), benzyl alcohol, polysorbate 80, diethylene glycol monoethyl ether (DEGEE), isopropyl myristate, ethanol, diisopropyl adipate, lactate C. 12~15 The pharmaceutical composition of embodiment 27 is provided, wherein the at least one selected from the group consisting of an alkyl, a viscosity enhancing agent, hydroxypropylcellulose, and PEG4000.

[0375] Example 29 provides the pharmaceutical composition of example 28, wherein the viscosity enhancing agent comprises a concentrated dispersion of a copolymer of acrylamide and sodium acryloyldimethyltaurate in isohexadecane.

[0376] Embodiment 30 provides the pharmaceutical composition of embodiment 29, wherein at least one of the following is present: (a) Compound 111, constituting about 0.1% to about 10.0% (w / w) of the pharmaceutical composition; (b) water, constituting about 10% to about 15% (w / w) of the pharmaceutical composition; (c) PEG 400 constituting about 20% to about 40% (w / w) of the pharmaceutical composition; (d) PEG300 constituting about 35% to about 60% (w / w) of the pharmaceutical composition; (e) PG constituting about 5% to about 15% (w / w) of the pharmaceutical composition; (f) benzyl alcohol, constituting from about 0.1% to about 5% (w / w) of the pharmaceutical composition; (g) polysorbate 80, constituting about 1% to about 10% (w / w) of the pharmaceutical composition; (h) a DEGEE constituting about 1% to about 15% (w / w) of the pharmaceutical composition; (i) isopropyl myristate, constituting about 0.1% to about 5% (w / w) of the pharmaceutical composition; (j) ethanol constituting about 5% to about 15% (w / w) of the pharmaceutical composition; (k) diisopropyl adipate, constituting about 5% to about 15% (w / w) of the pharmaceutical composition; (l) Lactic acid C, constituting about 5% to about 15% (w / w) of the pharmaceutical composition 12~15 Alkyl; (m) a viscosity enhancing agent comprising about 1% to about 10% (w / w) of the pharmaceutical composition; (n) hydroxypropylcellulose, constituting about 0.1% to about 5% (w / w) of the pharmaceutical composition; and (o) PEG 4000, constituting about 5% to about 15% (w / w) of the pharmaceutical composition.

[0377] Embodiment 31 provides the pharmaceutical composition of embodiment 30, consisting essentially of 111 (about 1.0% w / w), PEG400 (about 24.3% w / w), PEG300 (about 40.0% w / w), PG (about 10.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), isopropyl myristate (about 2.0% w / w), and PEG4000 (about 10% w / w).

[0378] Embodiment 32 provides the pharmaceutical composition of embodiment 30, consisting essentially of 111 (about 1.0% w / w), PEG400 (about 23.6% w / w), PEG300 (about 40.0% w / w), PG (about 10.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), and PEG4000 (about 10.0% w / w).

[0379] Embodiment 33 provides the pharmaceutical composition of embodiment 30, consisting essentially of 111 (about 1.0% w / w), PEG400 (about 34.0% w / w), PEG300 (about 40.0% w / w), PG (about 10% w / w), benzyl alcohol (about 2.0% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), isopropyl myristate (about 2.0% w / w), and hydroxypropylcellulose (about 1.0% w / w).

[0380] Embodiment 34 provides the pharmaceutical composition of embodiment 30, consisting essentially of 111 (about 1.0% w / w), water (about 13.3% w / w), PEG300 (about 57.0% w / w), PG (about 10% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), isopropyl myristate (about 2.0% w / w), and a viscosity enhancing agent (about 4% w / w).

[0381] Embodiment 35 is a mixture of 111 (about 1.0% w / w), PEG300 (about 48.0% w / w), PG (about 10% w / w), benzyl alcohol (about 1.5% w / w), DEGEE (about 10% w / w), ethanol (about 8.5% w / w), diisopropyl adipate (about 10% w / w), lactate C 12~15 The pharmaceutical composition of embodiment 30 is provided, consisting essentially of alkyl (about 10% w / w), and hydroxypropyl cellulose (about 1.0% w / w).

[0382] Embodiment 36 provides the pharmaceutical composition of embodiment 25 or 26, wherein at least one compound is compound 99.

[0383] Aspect 37 is a pharmaceutical composition comprising the at least one pharma- ceutically acceptable excipient selected from the group consisting of water, PEG400, PG, benzyl alcohol, polysorbate 80, DEGEE, isopropyl myristate, ethanol, diisopropyl adipate, and lactate C. 12~15The pharmaceutical composition of embodiment 36 is provided, wherein the at least one selected from the group consisting of alkyl, dimethyl isosorbide, PEG40 hydrogenated castor oil (HCO), hydroxypropyl cellulose, and PEG4000.

[0384] Embodiment 38 provides the pharmaceutical composition of embodiment 36 or 37, wherein at least one of the following is present: (a) Compound 99, constituting about 0.1% to about 10.0% (w / w) of the pharmaceutical composition; (b) water, constituting about 1% to about 10% (w / w) of the pharmaceutical composition; (c) PEG 400 constituting about 25% to about 35% (w / w) of the pharmaceutical composition; (d) PG constituting about 10% to about 30% (w / w) of the pharmaceutical composition; (e) benzyl alcohol, constituting about 0.1% to about 5% (w / w) of the pharmaceutical composition; (f) polysorbate 80, constituting about 1% to about 10% (w / w) of the pharmaceutical composition; (g) a DEGEE constituting about 10% to about 50% (w / w) of the pharmaceutical composition; (h) isopropyl myristate, constituting about 0.1% to about 5% (w / w) of the pharmaceutical composition; (i) ethanol constituting about 20% to about 35% (w / w) of the pharmaceutical composition; (j) diisopropyl adipate, constituting about 5% to about 15% (w / w) of the pharmaceutical composition; (k) lactic acid C, constituting about 1% to about 15% (w / w) of the pharmaceutical composition 12~15 Alkyl; (l) dimethylisosorbide, constituting about 5% to about 15% (w / w) of the pharmaceutical composition; (m) PEG40 HCO, constituting about 1% to about 10% (w / w) of the pharmaceutical composition; (n) hydroxypropylcellulose, constituting about 0.1% to about 5% (w / w) of the pharmaceutical composition; and (o) PEG 4000, constituting about 5% to about 15% (w / w) of the pharmaceutical composition.

[0385] Embodiment 39 provides the pharmaceutical composition of embodiment 38, consisting essentially of PEG 99 (about 1.0% w / w), PEG 400 (about 28.3% w / w), PG (about 20.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 25.0% w / w), isopropyl myristate (about 2.0% w / w), dimethyl isosorbide (about 10.0% w / w), PEG 40 HCO (about 5.0% w / w), and hydroxypropyl cellulose (about 1.0% w / w).

[0386] Embodiment 40 provides the pharmaceutical composition of embodiment 38, consisting essentially of PEG 99 (about 1.0% w / w), PEG 400 (about 30.30% w / w), PG (about 20.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate (about 5.0% w / w), DEGEE (about 25.0% w / w), dimethyl isosorbide (about 10.0% w / w), PEG 40 HCO (about 5.0% w / w), and hydroxypropyl cellulose (about 1.0% w / w).

[0387] Embodiment 41 provides the pharmaceutical composition of embodiment 38, consisting essentially of PEG 99 (about 1.0% w / w), PEG400 (about 19.30% w / w), PG (about 20.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate (about 5.0% w / w), DEGEE (about 25.0% w / w), isopropyl myristate (about 2.0% w / w), dimethyl isosorbide (about 10.0% w / w), PEG40 HCO (about 5.0% w / w), and PEG4000 (about 10.0% w / w).

[0388] Embodiment 42 is a mixture of 99 (about 1.0% w / w), DEGEE (about 40.0% w / w), ethanol (about 28.0% w / w), diisopropyl adipate (about 10.0% w / w), lactate C 12~15 The pharmaceutical composition of embodiment 38 is provided, consisting essentially of alkyl (about 10.0% w / w), dimethyl isosorbide (about 10.0% w / w), and hydroxypropyl cellulose (about 1.0% w / w).

[0389] Embodiment 43 is a mixture of 99 (about 1.0% w / w), water (about 5.0% w / w), DEGEE (about 42.5% w / w), ethanol (about 25.0% w / w), diisopropyl adipate (about 10.0% w / w), lactate C 12~15 The pharmaceutical composition of embodiment 38 is provided, consisting essentially of alkyl (about 5.0% w / w), dimethyl isosorbide (about 10.0% w / w), and hydroxypropyl cellulose (about 1.5% w / w).

[0390] Embodiment 44 provides the pharmaceutical composition of any one of embodiments 25 to 43, formulated for topical administration.

[0391] Embodiment 45 provides the pharmaceutical composition of embodiment 44, wherein the topical formulation comprises a gel or an ointment.

[0392] Embodiment 46 provides a method of treating, ameliorating, and / or preventing an orthopoxvirus infection in a human subject in need thereof, comprising the steps of: A therapeutically effective amount of at least one pharmaceutical composition according to any one of embodiments 25 to 45, and / or a compound of formula (II): administering to the subject a compound of the formula: TIFF2024536861000154.tif26128, or a salt, solvate, enantiomer, diastereoisomer, geometric isomer, or tautomer thereof; During the ceremony, X is CR 1 or N; Y is CR 2 or N; R 1 is H, optionally substituted C1-C6 alkyl, -C(=O)NR 6 R 6 , -C(=O)OR 6 , or R 8 and; R 2 is H or optionally substituted C1-C6 alkyl; R 3 H, -CN, -C(=O)OR 6 , -C(=O)NR 6 R6 , optionally substituted phenyl, or optionally substituted C1-C6 alkyl; R 4 -C(=O)OR 6 or R 8 and; R 5 is, at each occurrence, independently, optionally substituted C1-C6 alkyl or optionally substituted phenyl; R 6 is independently at each occurrence H or an optionally substituted C1-C6 alkyl; or two R's 6 together with the N atom to which they are both attached form an optionally substituted 4- to 7-membered heterocyclyl; R 8 is -C(=O)NH(optionally substituted acyl), -N(optionally substituted acyl)C(=O)R 7 , -NR 6 C(=O)R 7 or -NR 6 C(=O)NR 6 R 7 and; R 7 each occurrence independently represents optionally substituted C1-C6 alkyl, optionally substituted cycloalkyl, CH(optionally substituted heterocyclyl)(R 5 ), CH(R 5 )(R 5 ), or an optionally substituted 4- to 7-membered heterocyclyl; or R 6 and R 7 together with the N atom to which they are both attached form an optionally substituted 4- to 7-membered heterocyclyl; However, (I) is R 8 Contains one The above process.

[0393] Example 47 provides the method of example 46, wherein the orthopoxvirus infection is caused by a virus selected from the group consisting of Molluscum contagiosum virus (MCV), amerpoxvirus, cowpox virus, mousepox virus, horsepoxvirus, monkeypox virus, raccoonpox virus, tanapox virus, variola (smallpox) virus, yokapoxvirus, cervidopoxvirus (deerpox), avipoxvirus (fowlpox), capripoxvirus (goatpox), leporipoxvirus (myxoma virus), parapoxvirus (orph virus), suipoxvirus (swinepox), and yatapoxvirus (yaba-like disease virus).

[0394] Example 48 provides the method of example 47, wherein the orthopoxvirus infection is caused by molluscum contagiosum virus (MCV).

[0395] Example 49 provides the method of Example 46, wherein the compound or composition is applied to the skin of the subject.

[0396] Example 50 provides the method of Example 46, wherein the compound or composition is applied to at least one MCV lesion on the skin of the subject.

[0397] Example 51 provides the method of example 46, wherein the at least one compound or composition is formulated as a topical pharmaceutical composition.

[0398] Example 52 provides the method of example 51, wherein the topical pharmaceutical composition comprises a gel or ointment.

[0399] The disclosures of all patents, patent applications, and publications cited herein are incorporated herein by reference in their entirety. Although the present invention has been disclosed with reference to certain embodiments, it is apparent that other embodiments and modifications of the present invention may be made by others skilled in the art without departing from the true spirit and scope of the invention. It is intended that the appended claims be construed to include all such embodiments and equivalent modifications.

Claims

1. A compound of formula (I) or a salt, solvate, enantiomer, diastereoisomer, geometric isomer, or tautomer thereof: During the ceremony, X is CR 1 or N; Y is CR 2 or N; R 1 is H, optionally substituted C 1 ~C 6 Alkyl, -C(=O)NR 6 R 6 , -C(=O)OR 6 , or R 8 and R 2 is H or optionally substituted C 1 ~C 6 is alkyl; R 3 is H, -CN, -C(=O)OR 6 , -C(=O)NR 6 R 6 , optionally substituted phenyl, or optionally substituted C 1 ~C 6 is alkyl; R 4 is -C(=O)OR 6 or R 8 and R 5 each occurrence independently represents an optionally substituted C 1 ~C 6 alkyl or optionally substituted phenyl; R 6 is, in each occurrence, independently H or optionally substituted C 1 ~C 6 Is it alkyl? or two R's 6 together with the N atom to which they are both attached form an optionally substituted 4- to 7-membered heterocyclyl; R 8 is -C(=O)NH(optionally substituted acyl), -N(optionally substituted acyl)C(=O)R 7 , -NR 6 C(=O)R 7 or -NR 6 C(=O)NR 6 R 7 and R 7 each occurrence independently represents an optionally substituted C 1 ~C 6 alkyl, optionally substituted cycloalkyl, CH(optionally substituted heterocyclyl)(R 5 ), CH(R 5 )(R 5 ), or an optionally substituted 4- to 7-membered heterocyclyl; or R 6 and R 7 together with the N atom to which they are both attached form an optionally substituted 4- to 7-membered heterocyclyl; However, (I) is R 8 and With the proviso that the compound is not selected from the group consisting of the following compounds: 1, 2, 4, 5, 8, 9, 15, 16, 17, 19, 24, 26, 29, 31, 40, 66, 72, 76, 77, 78, 83, 84, and 86; 。

2. X is CR 1 and Y is CR 2 2. The compound of claim 1, wherein:

3. 2. The compound of claim 1, wherein X is N and Y is N.

4. X is CR 1 and Y is N.

5. X is N and Y is CR 2 2. The compound of claim 1, wherein:

6. The compound of claim 1 selected from the group consisting of:

7. R 4 R 8 7. The compound of any one of claims 1 to 6, wherein

8. The compound of any one of claims 1 to 6, wherein at least one of the following is true: R 8 Ha-NR 6 C(=O)R 7 and R 6 is H and R 7 is CH(CH 2 CH 3 )Ph; R 8 Ha-NR 6 C(=O)R 7 and R 6 is H and R 7 is CH(CH 2 CH 3 )(4-F-Ph); R 8 Ha-NR 6 C(=O)NR 6 R 7 and R 6 is H and NR 6 R 7 is 4-phenyl-piperazin-1-yl; R 8 Ha-NR 6 C(=O)NR 6 R 7 and R 6 is H and NR 6 R 7 is 4-(2-pyridyl)-piperazin-1-yl; R 3 is —C(═O)OR 6 and R 6 is CH 3 ; R 3 is —C(═O)OR 6 and R 6 is CH 2 CH 3 ; or R 3 is —C(═O)NR 6 R 6 , where NR 6 R 6 is NH(CH 2 -aryl), wherein the aryl is selected from the group consisting of phenyl, 4-fluorophenyl, 4-chlorophenyl, and 4-trifluoromethylphenyl.

9. R 1 and R 4 One of the The compound of any one of claims 1 to 6, selected from the group consisting of: During the ceremony, R a1 and R a2 are, if present, each independently H and optionally substituted C 1 ~C 6 selected from the group consisting of alkyl; R b1 , R b2 , R b3 , R b4 , and R b5 are, if present, each independently H, optionally substituted C 1 ~C 6 Alkyl, optionally substituted C 1 ~C 6 Alkoxy, optionally substituted C 1 ~C 6 Haloalkyl, halogen, CN, and NO 2 selected from the group consisting of R b1 , R b2 , R b3 , R b4 , and R b5 Two vicinal substituents selected from the group consisting of: 2 ~C 10 may form a heterocycloalkyl; A 1 is optionally substituted C 2 ~C 10 Heteroaryl and optionally substituted C 2 ~C 10 heterocycloalkyl; G 1 is a bond and C(R c6 )(R c7 ) selected from the group consisting of; G 2 is a bond and C(R c8 )(R c9 ) selected from the group consisting of; G 3 is a bond and C(R c10 )(R c11 ) selected from the group consisting of; R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 are, if present, each independently H, optionally substituted C 1 ~C 6 Alkyl, optionally substituted C 1 ~C 6 Alkoxy, optionally substituted C 1 ~C 6 selected from the group consisting of haloalkyl, halogen; R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 Two vicinal substituents selected from the group consisting of: 6 ~C 10 may form an aryl; Z 1 is N and CR c9 and Z 2 is N and CR b5 is selected from the group consisting of:

10. The compound of claim 9, wherein one of the following applies: (a)R a1 is H and R a2 is ethyl; (b)R a1 is ethyl, and R a2 is H; (c) each of R b1 , R b2 , R b3 , R b4 , and R b5 , if present, is independently selected from the group consisting of H, Me, OMe, F, Cl, CF 3 , CN, and NO 2 ; or (d) Each of R c1 , R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 , when present, is independently selected from the group consisting of H and Ph.

11. The compound of claim 10, wherein at least one of the following is true: (a)G 1 , G 2 , and G 3 None of the are bonds; (b) G 1 , G 2 , and G 3 One of them is a bond; (c)G 1 , G 2 , and G 3 two of which are bonds; and (d)G 1 , G 2 , and G 3 Each of these is a bond.

12. A 1 but 11. The compound of claim 10, selected from the group consisting of:

13. The compound of claim 10, wherein at least one of the following is true: (a) R 2 is optionally substituted C 1 ~C 6 is alkyl, R 3 is C(=O)OR 6 and R 1 and R 4 One of the and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF 3 , CN, and NO 2 or R b1 , R b2 , R b3 , R b4 , and R b5 two vicinal substituents selected from the group consisting of: taken together to form methylenedioxy; (b) R 3 is optionally substituted C 1 ~C 6 is alkyl, R 2 is C(=O)OR 6 and R 1 and R 4 One of the and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF 3 , CN, and NO 2 or R b1 , R b2 , R b3 , R b4 , and R b5 two vicinal substituents selected from the group consisting of: taken together to form methylenedioxy; (c) R 2 is optionally substituted C 1 ~C 6 is alkyl, R 3 is CN, R 1 and R 4 One of the and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF 3 , CN, and NO 2 or R b1 , R b2 , R b3 , R b4 , and R b5 two vicinal substituents selected from the group consisting of: taken together to form methylenedioxy; (d) R 2 is C(=O)OR 6 and R 1 and R 4 One of the and R 3 and R 1 or R 4 C 1 ~C 6 Not more than one can be alkyl; (e) R 1 is H, R 2 is H, R 3 is -C(=O)NR 6 R 6 and R 6 At most one occurrence of is H; (f) R 1 teeth and R 2 is methyl, R 3 is H, and R 4 is C(=O)O(C 1 alkyl), C(=O)O(optionally substituted C 3 alkyl), C(=O)O(optionally substituted C 4 alkyl), C(=O)O(optionally substituted C 5 alkyl), and C(=O)O (optionally substituted C 6 alkyl); (g) R 4 teeth and R 3 is methyl, R 2 is H, and R 1 is C(=O)O(C 1 alkyl), C(=O)O(optionally substituted C 3 alkyl), C(=O)O(optionally substituted C 4 alkyl), C(=O)O(optionally substituted C 5 alkyl), and C(=O)O (optionally substituted C 6 alkyl); (h) R 1 is C(=O)NH 2 and R 4 teeth and G 1 , G 2 , and G 3 At most one of the is a bond, and R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 A pair of vicinal substituents selected from the group consisting of, together with the atoms to which they are attached, is C 6 ~C 10 Forming an aryl; (i) R 1 is C(=O)NH 2 and R 4 teeth and G 1 , G 2 , and G 3 At most one of the is a bond, and R c2 , R c3 , R c4 , R c5 , R c6 , R c7 , R c8 , R c9 , R c10 , and R c11 A pair of vicinal substituents selected from the group consisting of, together with the atoms to which they are attached, is C 6 ~C 10 Forming an aryl; (j) Y is N, X is CR 1 and R 1 and R 4 One of the and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF 3 , CN, and NO 2 or R b1 , R b2 , R b3 , R b4 , and R b5 two vicinal substituents selected from the group consisting of: taken together to form methylenedioxy; (k) X is N, Y is CR 2 and R 4 teeth and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of Me, OMe, F, Cl, CF 3 , CN, and NO 2 or R b1 , R b2 , R b3 , R b4 , and R b5 two vicinal substituents selected from the group consisting of: taken together to form methylenedioxy; (l) X is N, Y is N, R 4 teeth and Z 1 is CR c9 and (m) X is N, Y is N, R 4 teeth and Z 2 is CR b5 and R b1 , R b2 , R b3 , R b4 , and R b5 At least one selected from the group consisting of halogen, C 1 ~C 6 Alkyl, NO 2 and CN.

14. R 1 がH、Me、C(=O)NH 2 、C(=O)NMe 2 、C(=O)NEt 2 、C(=O)OEt、C(=O)OMe、C(=O)OEt、C(=O)Hot-Bu、C(=O)O(CH 2 ) 2 NH 2 、C(=O)NH(CH 2 ) 3 NH 2 、 7. The compound of any one of claims 1 to 6, selected from the group consisting of:

15. R 4 がMe、C(=O)NH 2 、C(=O)NMe 2 、C(=O)NEt 2 、C(=O)OEt、C(=O)OMe、C(=O)OEt、C(=O)Hot-Bu、C(=O)O(CH 2 ) 2 NH 2 、C(=O)NH(CH 2 ) 3 NH 2 、 7. The compound of any one of claims 1 to 6, selected from the group consisting of:

16. R 2 7. The compound of any one of claims 1 to 6, wherein is selected from the group consisting of H, Me, and Et.

17. R 3 is H, Me, Et, C(=O)OMe, C(=O)OEt, C(=O)NH 2 , CN, Ph, 4-trifluoromethylphenyl, 4-fluorophenyl, 7. The compound of any one of claims 1 to 6, selected from the group consisting of:

18. The following compounds 22, 32, 33, 34, 35, 36, 39, 47, 50, 57, 68, 75, 82, 87, 89, 90, 95, 96, 97, 98, 99, 106, 109, 110, 111, 112, 114, 115, 116, 118, 119, 123, 124, 127, 130, 131, 132, 134, 135, 144, 153, 1 10. The compound of any one of claims 1 to 6, selected from the group consisting of: 54, 155, 159, 160, 161, 162, 163, 165, 167, 168, 169, 170, 172, 173, 174, 175, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, and 191; 。 19. The compound of any one of claims 1 to 6, wherein at least one of the following is true: (a) each occurrence of alkyl or cycloalkyl is independently C 1 ~C 6 Alkyl, halo, carbonyl (C=O), -OR, optionally substituted phenyl, optionally substituted heteroaryl, optionally substituted heterocyclyl, -N(R)(R), -N(R)-(C=O)R, -C(=O)R, -C(=O)(optionally substituted phenyl), -C(=O)(optionally substituted heteroaryl), -C(=O)N(R)(R), -C(=O)(CH 2 ) 0～3 OR, -S(=O) 2 R, and -SO 2 and optionally substituted with at least one substituent selected from the group consisting of N(R)(R), where R, at each occurrence, is independently selected from H, C, 1 ~C 6 Alkyl, and C 3 ~C 8 cycloalkyl; and (b) phenyl, in each occurrence, is optionally substituted with at least one substituent selected from the group consisting of C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 haloalkoxy, halo, -CN, -OR, -N(R)(R), -NO2, -S(=O)2N(R)(R), acyl, and C1-C6 alkoxycarbonyl, where R, in each occurrence, is independently selected from the group consisting of H, C1-C6 alkyl, and C3-C8 cycloalkyl; Preferably, phenyl, at each occurrence, is optionally substituted with at least one substituent independently selected from the group consisting of C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 haloalkoxy, halo, -CN, -OR, -N(R)(R), and C1-C6 alkoxycarbonyl, and R, at each occurrence, is independently selected from the group consisting of H, C1-C6 alkyl, and C3-C8 cycloalkyl.

20. A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 6 and at least one pharmaceutically acceptable excipient.

21. The following compounds 1, 2, 4, 5, 8, 9, 15, 16, 17, 19, 24, 26, 29, 31, 40, 66, 72, 76, 77, 78, 83, 84, 86, 99, and 111: and at least one pharmaceutically acceptable excipient.

22. The following compound 111: and at least one pharmaceutically acceptable excipient.

23. At least one pharmaceutically acceptable excipient is selected from the group consisting of water, polyethylene glycol (PEG) 400, PEG 300, propylene glycol (PG), benzyl alcohol, polysorbate 80, diethylene glycol monoethyl ether (DEGEE), isopropyl myristate, ethanol, diisopropyl adipate, and lactic acid C 12～15 at least one selected from the group consisting of alkyl, thickener, hydroxypropyl cellulose, and PEG 4000; 23. The pharmaceutical composition of claim 22, wherein the viscosity increasing agent preferably comprises a concentrated dispersion of a copolymer of acrylamide and sodium acryloyldimethyltaurate in isohexadecane.

24. 24. The pharmaceutical composition of claim 23, wherein at least one of the following is present: (a) Compound 111, comprising about 0.1% to about 10.0% (w / w) of the pharmaceutical composition; (b) water, comprising about 10% to about 15% (w / w) of the pharmaceutical composition; (c) PEG400 comprising about 20% to about 40% (w / w) of the pharmaceutical composition; (d) PEG300 comprising about 35% to about 60% (w / w) of the pharmaceutical composition; (e) PG, comprising about 5% to about 15% (w / w) of the pharmaceutical composition; (f) benzyl alcohol, comprising about 0.1% to about 5% (w / w) of the pharmaceutical composition; (g) polysorbate 80, comprising about 1% to about 10% (w / w) of the pharmaceutical composition; (h) a DEGEE comprising about 1% to about 15% (w / w) of the pharmaceutical composition; (i) isopropyl myristate, comprising about 0.1% to about 5% (w / w) of the pharmaceutical composition; (j) ethanol comprising about 5% to about 15% (w / w) of the pharmaceutical composition; (k) diisopropyl adipate, comprising about 5% to about 15% (w / w) of the pharmaceutical composition; (l) lactic acid C, constituting about 5% to about 15% (w / w) of the pharmaceutical composition 12～15 Alkyl; (m) a viscosity enhancing agent comprising about 1% to about 10% (w / w) of the pharmaceutical composition; (n) hydroxypropyl cellulose, comprising about 0.1% to about 5% (w / w) of the pharmaceutical composition; and (o) PEG 4000 comprising about 5% to about 15% (w / w) of the pharmaceutical composition.

25. The pharmaceutical composition of claim 24, consisting essentially of: (a) Compound 111 (about 1.0% w / w), PEG400 (about 24.3% w / w), PEG300 (about 40.0% w / w), PG (about 10.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), isopropyl myristate (about 2.0% w / w), and PEG4000 (about 10% w / w); (b) Compound 111 (about 1.0% w / w), PEG400 (about 23.6% w / w), PEG300 (about 40.0% w / w), PG (about 10.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), and PEG4000 (about 10.0% w / w); (c) Compound 111 (about 1.0% w / w), PEG400 (about 34.0% w / w), PEG300 (about 40.0% w / w), PG (about 10% w / w), benzyl alcohol (about 2.0% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), isopropyl myristate (about 2.0% w / w), and hydroxypropyl cellulose (about 1.0% w / w). (d) Compound 111 (about 1.0% w / w), water (about 13.3% w / w), PEG300 (about 57.0% w / w), PG (about 10% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 5.0% w / w), isopropyl myristate (about 2.0% w / w), and a viscosity increasing agent (about 4% w / w); or (e) Compound 111 (about 1.0% w / w), PEG300 (about 48.0% w / w), PG (about 10% w / w), benzyl alcohol (about 1.5% w / w), DEGEE (about 10% w / w), ethanol (about 8.5% w / w), diisopropyl adipate (about 10% w / w), C 12-15 alkyl lactate (about 10% w / w), and hydroxypropyl cellulose (about 1.0% w / w).

26. Compound 99 below: and at least one pharmaceutically acceptable excipient, Optionally, the pharmaceutical composition wherein the at least one pharmaceutically acceptable excipient is at least one selected from the group consisting of water, PEG400, PG, benzyl alcohol, polysorbate 80, DEGEE, isopropyl myristate, ethanol, diisopropyl adipate, C12-15 alkyl lactate, dimethyl isosorbide, PEG40 hydrogenated castor oil (HCO), hydroxypropyl cellulose, and PEG4000.

27. 27. The pharmaceutical composition of claim 26, wherein at least one of the following is present: (a) Compound 99, comprising about 0.1% to about 10.0% (w / w) of the pharmaceutical composition; (b) water, comprising about 1% to about 10% (w / w) of the pharmaceutical composition; (c) PEG 400 comprising about 25% to about 35% (w / w) of the pharmaceutical composition; (d) PG, comprising about 10% to about 30% (w / w) of the pharmaceutical composition; (e) benzyl alcohol, comprising about 0.1% to about 5% (w / w) of the pharmaceutical composition; (f) polysorbate 80, comprising about 1% to about 10% (w / w) of the pharmaceutical composition; (g) a DEGEE comprising about 10% to about 50% (w / w) of the pharmaceutical composition; (h) isopropyl myristate, comprising about 0.1% to about 5% (w / w) of the pharmaceutical composition; (i) ethanol, comprising about 20% to about 35% (w / w) of the pharmaceutical composition; (j) diisopropyl adipate, comprising about 5% to about 15% (w / w) of the pharmaceutical composition; (k) lactic acid C, constituting about 1% to about 15% (w / w) of the pharmaceutical composition 12～15 Alkyl; (l) dimethylisosorbide, comprising about 5% to about 15% (w / w) of the pharmaceutical composition; (m) PEG40 HCO, comprising about 1% to about 10% (w / w) of the pharmaceutical composition; (n) hydroxypropyl cellulose, comprising about 0.1% to about 5% (w / w) of the pharmaceutical composition; and (o) PEG 4000 comprising about 5% to about 15% (w / w) of the pharmaceutical composition.

28. The pharmaceutical composition of claim 26, consisting essentially of: (a) Compound 99 (about 1.0% w / w), PEG400 (about 28.3% w / w), PG (about 20.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate 80 (about 5.0% w / w), DEGEE (about 25.0% w / w), isopropyl myristate (about 2.0% w / w), dimethyl isosorbide (about 10.0% w / w), PEG40 HCO (about 5.0% w / w), and hydroxypropyl cellulose (about 1.0% w / w); (b) Compound 99 (about 1.0% w / w), PEG400 (about 30.30% w / w), PG (about 20.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate (about 5.0% w / w), DEGEE (about 25.0% w / w), dimethyl isosorbide (about 10.0% w / w), PEG40 HCO (about 5.0% w / w), and hydroxypropyl cellulose (about 1.0% w / w); (c) Compound 99 (about 1.0% w / w), PEG400 (about 19.30% w / w), PG (about 20.0% w / w), benzyl alcohol (about 2.7% w / w), polysorbate (about 5.0% w / w), DEGEE (about 25.0% w / w), isopropyl myristate (about 2.0% w / w), dimethyl isosorbide (about 10.0% w / w), PEG40 HCO (about 5.0% w / w), and PEG4000 (about 10.0% w / w); (d) Compound 99 (about 1.0% w / w), DEGEE (about 40.0% w / w), ethanol (about 28.0% w / w), diisopropyl adipate (about 10.0% w / w), C 12-15 alkyl lactate (about 10.0% w / w), dimethyl isosorbide (about 10.0% w / w), and hydroxypropyl cellulose (about 1.0% w / w); or (e) Compound 99 (about 1.0% w / w), water (about 5.0% w / w), DEGEE (about 42.5% w / w), ethanol (about 25.0% w / w), diisopropyl adipate (about 10.0% w / w), C12-15 alkyl lactate (about 5.0% w / w), dimethyl isosorbide (about 10.0% w / w), and hydroxypropyl cellulose (about 1.5% w / w).

29. 1. A pharmaceutical composition formulated for topical administration, comprising:

29. The pharmaceutical composition of any one of claims 22, 24, and 28, wherein the topical formulation preferably comprises a gel or an ointment.

30. 1. A pharmaceutical composition for treating, ameliorating, and / or preventing an orthopoxvirus infection in a human subject in need thereof, the pharmaceutical composition comprising at least one compound of claim 1 and / or a compound of formula (II): or a salt, solvate, enantiomer, diastereoisomer, geometric isomer or tautomer thereof, During the ceremony, X is CR 1 or N; Y is CR 2 or N; R 1 is H, optionally substituted C 1 ~C 6 Alkyl, -C(=O)NR 6 R 6 , -C(=O)OR 6 , or R 8 and R 2 is H or optionally substituted C 1 ~C 6 is alkyl; R 3 is H, -CN, -C(=O)OR 6 , -C(=O)NR 6 R 6 , optionally substituted phenyl, or optionally substituted C 1 ~C 6 is alkyl; R 4 is -C(=O)OR 6 or R 8 and R 5 each occurrence independently represents an optionally substituted C 1 ~C 6 alkyl or optionally substituted phenyl; R 6 is, in each occurrence, independently H or optionally substituted C 1 ~C 6 Is it alkyl? or two R's 6 together with the N atom to which they are both attached form an optionally substituted 4- to 7-membered heterocyclyl; R 8 is -C(=O)NH(optionally substituted acyl), -N(optionally substituted acyl)C(=O)R 7 , -NR 6 C(=O)R 7 or -NR 6 C(=O)NR 6 R 7 and R 7 each occurrence independently represents an optionally substituted C 1 ~C 6 alkyl, optionally substituted cycloalkyl, CH(optionally substituted heterocyclyl)(R 5 ), CH(R 5 )(R 5 ), or an optionally substituted 4- to 7-membered heterocyclyl; or R 6 and R 7 together with the N atom to which they are both attached form an optionally substituted 4- to 7-membered heterocyclyl; However, (II) is R 8 Contains one Preferably, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient. Pharmaceutical compositions.

31. the orthopoxvirus infection is caused by a virus selected from the group consisting of Molluscum contagiosum virus (MCV), amerpoxvirus, cowpox virus, mousepox virus, horsepox virus, monkeypox virus, raccoonpox virus, tanapox virus, variola (smallpox) virus, yokapoxvirus, cervidopoxvirus (deerpox), avipoxvirus (fowlpox), capripoxvirus (goatpox), leporipoxvirus (myxoma virus), parapoxvirus (orph virus), suipoxvirus (swinepox), and yatapoxvirus (yaba-like disease virus); 31. The pharmaceutical composition of claim 30, wherein the orthopoxvirus infection is preferably caused by molluscum contagiosum virus (MCV).

32. The pharmaceutical composition of claim 30 or 31, wherein at least one of the following applies: (a) applying the pharmaceutical composition to the skin of the subject; (b) applying the pharmaceutical composition to at least one MCV lesion on the skin of the subject; (c) the pharmaceutical composition is formulated as a topical pharmaceutical composition; and (d) the pharmaceutical composition is formulated as a topical pharmaceutical composition, the topical pharmaceutical composition comprising a gel or an ointment;

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