DERIVATIVES

The present invention relates to 3-(phenylsulfonyl)-[l,2,3] triazolo[l,5a]quinazolin- 5(4H)-one derivatives and pharmaceutical compositions thereof as well as to their uses in methods of reducing the virulence of bacteria (preferably Staphylococcus aureus') that express accessory gene regulator A (AgrA) or an ortholog of AgrA; in methods of inhibition of the quorum sensing in bacteria, preferably in 5. aureus, and in methods for preventing or treating diseases caused or exacerbated by bacteria, preferably by 5. aureus, including but not limited to skin or lung infections, atopic dermatitis, Netherton syndrome, and/or psoriasis in a subject. Thus, the present invention relates to anti-virulence compositions and methods for treatment, amelioration and/or prevention of diseases caused or exacerbated by bacteria, preferably by 5. aureus, and more particularly to compositions and methods for reducing the virulence of bacteria that express AgrA or an ortholog of AgrA, preferably AgrA.

RELATED ART

Staphylococcus aureus is both a human commensal and a notorious opportunistic pathogen causing serious community-acquired and hospital-acquired infections. 5. aureus is capable of causing a vast array of infections ranging from mild superficial skin infections to severe systemic life-threatening conditions such as endocarditis, pneumonia or sepsis (Lee AS, et al. (2018) Nat. Rev. Dis. Primers, Vol 4, Article 18033 pp. 1-23. Moreover, S. aureus has also been implicated in contributing to allergic skin conditions such as atopic dermatitis (Geoghegan JA, et al. (2018) Trends Microbiol. 26(6):484-497) and Netherton syndrome (Williams MR, et al. (2020) Cell reports 30 (9): 2923-2933). The success of 5. aureus to cause such a variety of diseases is a consequence of the extensive arsenal of virulence factors produced combined with P-lactam resistance and, for most clones, resistance to other antibiotic classes. Clinically relevant antibiotic resistance has evolved against virtually every antibiotic deployed, while the discovery and development of novel antibiotic classes is lagging behind, provoking the antibiotic resistance crisis we are facing today. Consequently, alternative strategies to treat or prevent 5. m/ra./.s-mediated bacterial infections that are also efficacious against multidrug-resistant strains, such as the methicillin-resistant 5. aureus (MRSA), are needed (Dickey SW, et al. (2017) Nat. Rev. Drug Discov. 16(7):457-471).

One of these strategies is the anti-virulence approach by which only virulence- associated, but not survival/fitness-relevant traits are targeted. In contrast to common antibiotic therapies, anti- virulence drugs are not per se bacteriostatic (inhibiting bacterial growth) or bactericidal (killing bacteria). This approach focuses on disarming the pathogenic bacteria by blocking the expression or neutralizing their virulence factors, ultimately interfering with bacterial pathogenicity mechanisms and thereby promoting pathogen clearance by the host immune system. Because anti-virulence drugs do not interfere with essential mechanisms of bacterial growth and survival, they are supposed to alleviate the pressure on the pathogen to develop resistance. A further advantage is that specific anti- virulence drugs preserve the healthy host microbiota, and eventually even help to counteract microbial dysbiosis by tuning down the aggressiveness of pathogens such as 5. aureus. Importantly, anti-virulence approaches offer an increased repository of pharmacological targets, and thus the possibility of generating alternative antimicrobials with novel mode of action (Muhlen, S. & Dersch, P. (2016) Curr. Top. Microbiol. Immunol. 398:147-183).

How the virulence factors are regulated in 5. aureus'. The agr operon is a bacterial quorum-sensing system that controls cell-density dependent virulence factor expression in 5. aureus. It consists of two divergent promoters, P2 and P3, where P2 is responsible for producing the components of the quorum-sensing system (AgrB, AgrD, AgrC, and AgrA; see FIG 1). The precursor peptide AgrD is processed by AgrB to form the mature auto-inducing peptide (AIP) that is secreted across the bacterial membrane. AIP binds to the histidine-kinase AgrC and activates the transcriptional regulator AgrA by means of phosphorylation, driving the expression from P2 and P3. There are four allelic variants (types I-IV) of agr, each encoding a distinct AIP, which functions as specific ligand for the AgrC of its own cell, but as an inhibitor of other AgrC variants. P3 produces the agr effector molecule RNAIII that together with AgrA is responsible for transcriptional control of approximately 200 genes including multiple virulence factors and metabolic pathways involved in stationary phase growth (Khan, BA, et al. (2015) Expert Opin. Investig. Drugs 24(5):689-704). Examples for AgrA-regulated virulence factors are cell-surface associated proteins, such as protein A (SpA) and fibronectin- binding proteins, secreted toxins such as a-hemolysin/a-toxin (Hla), 5-hemolysin (Hid), phenol-soluble-modulins (PSMs), Panton-Valentine leukocidin (PVL), leukotoxin E and D (LukED), leukotoxin G and H (LukGH), or secreted proteases such as SspA or aureolysin. Taken together, the numerous and multi-functional virulence determinants make S. aureus pathogenesis particularly complex and provide the pathogen with an arsenal of mechanisms to damage the host or circumvent and evade the host immune defenses. Important to note, most AgrA-regulated virulence factors are causative for the 5. aureus pathogenicity in skin and soft tissue infections (SSTIs), lung infections, and were also shown to contribute to chronic inflammatory skin diseases, such as atopic dermatitis (Oliveira D. etal. (201 ) /Aw7.s' 10: 252; Geoghegan JA, et al. (2018) Trends Microbiol. 26(6): 484-497) and Netherton syndrome (Williams MR, et al. (2020) Cell reports 30 (9): 2923-2933).

How to prevent virulence factor expression: Inhibition of expression of the central regulatory RNAIII combined with inhibition of PSMa production is believed to lead to apotent global reduction in levels of virulence factors (see FIG 1). Current strategies to suppress RNAIII expression can be grouped into different categories: (1) competitive inhibitors of histidine kinase AgrC; (2) RNAIII transcription inhibitors (precise mechanisms undetermined); and (3) inhibition of AgrA-P2/Ps interactions. Targeting AgrA has the advantage of blocking AgrA- dependent virulence factor expression on all four agr groups (Gordon CP, et al. (2013) J. Med. Chem. 56(4): 1389-404).

Recently, an AgrA inhibitor termed Savirin was found by Sully and colleagues in a screen of 24’087 compounds selected for inhibition of cyclic thiolactone peptide pheromone (AlP)-induced agr in the context of studies related to acute bacterial skin and soft tissue infections caused by S. aureus (Sully EK, et al. (2014) PLoS Pathog. 10(6): el004174). Savirin was shown to be a potent modulator of AgrA-regulated toxin gene transcription such as hla, psma and pvl across all four agr groups without affecting viability of S. aureus. Savirin inhibited exotoxin-induced red blood cell (RBC) lysis. No resistance was developed after multiple passages with Savirin. The molecule was shown to interfere with the AgrA-DNA interaction, preventing virulence gene upregulation. A significant reduction in abscess size and dermonecrosis was observed in mice infected with a MRSA USA300 type strain when Savirin was applied multiple times.

Recently, WO 2020/109350 described 3-(phenylsulfonyl)-[l,2,3]triazolo[l,5a]quina- zolin-5(4h)-one derivatives that are capable of reducing the virulence of bacteria such as 5. aureus by interacting with AgrA and inhibiting the expression of AgrA-regulated virulence factors, and their use in preventing or treating bacterial infections and/or diseases caused or exacerbated by bacteria, preferably by 5. aureus, including skin or lung infections, atopic dermatitis, or psoriasis.

SUMMARY OF THE INVENTION

The inventors have surprisingly found that the ability to inhibit AgrA-related virulence factors by compounds of WO 2020/109350 could be affected and reduced when tested in the presence of human serum (HS) or human serum albumin (HSA; the major component of human serum). Without wishing to be bound, the reduced ability to inhibit expression of AgrA-related virulence factors is thought to be due to nonspecific binding of the compounds to serum proteins (e.g., HSA). Such reduced efficacy in the presence of HS or HSA can limit the ability to inhibit AgrA-related virulence factors when administered (e.g., in an animal, preferably a mammal, more preferably a human). The present invention now provides a new class of compounds overcoming said drawbacks, and thus provides compounds that can reduce the virulence of bacteria, preferably that express AgrA and/or inhibit quorum sensing in bacteria, preferably in 5. aureus, in biologically relevant systems that include human serum or human serum albumin.

Thus, the inventors have surprisingly identified a new class of compounds able to interact with AgrA and inhibit the expression of AgrA-regulated virulence factors even in the presence of human serum (HS) or human serum albumin (HSA). The inventors transfected 5. aureus with a plasmid expressing the reporter gene lacZ under the control of the P3 promoter, which is regulated by AgrA. In the presence of the inventive compounds, the inventors found that production of the gene product of lacZ (i.e., B-galactosidase) was inhibited. Surprisingly, the inventive compounds retained AgrA inhibition activity even when the assay was performed in the presence of HSA.

The inventive compounds can thus inhibit the transcription of genes (preferably virulence factors) under the control of the P3 promoter (e.g., in bacteria, preferably in 5. aureus) in the presence of HS or HSA. This activity strongly suggests that the inventive compounds can be administered in biologically relevant systems (e.g., in animals, preferably mammals, more preferably humans) while still maintaining efficacy. Additional features and advantages of the present technology will be understood by those of skill in the art upon reading the Detailed Description of the Invention, below.

In a first aspect, the present invention provides a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein: R ¹ is independently selected from -H, halogen, -Ci-Ce alkyl, and -Ci-Ce alkoxy, wherein said alkyl and alkoxy is each optionally substituted with one or more R ¹¹ ;

R ³ is independently selected from -H, halogen, Ci-Ce alkyl, -Ci-Ce alkoxy, and -C3-C6 cycloalkyl, wherein said alkyl, alkoxy, and cycloalkyl is each optionally substituted by one or more R ¹¹ ;

R ^8A and R ^8B are each independently, at each occurrence, -H or -CH3;

R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C3-C8 cycloalkyl, -Ci-Ce alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, - Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkylene-(C6-Cio aryl), 5- to 10-membered heteroaryl, and -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, - (OCH ₂ -CH ₂ )I- ₂ -OH, -CI-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); and wherein said cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is each independently optionally substituted with one or more R ⁸⁵ ;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl;

R ⁸⁵ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, -OH, - N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, -Ce-Cio aryl, and oxo; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more R ⁸⁶ ;

R ⁸⁶ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, -OH, - Ci-Ce alkylene-OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl, and oxo;

R ¹¹ is independently, at each occurrence, halogen or -OH; m is independently 1 or 2; and n is independently 0 or 1.

In one aspect, the invention provides an inventive compound as described herein or a pharmaceutical composition comprising an inventive compound as described herein for use in a method of reducing the virulence of a bacteria, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA.

In one aspect, the invention provides an inventive compound as described herein or a pharmaceutical composition comprising an inventive compound as described herein for use in a method of inhibiting the quorum sensing in bacteria, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA.

In one aspect, the invention provides an inventive compound as described herein or a pharmaceutical composition comprising an inventive compound as described herein for use in a method of preventing or treating diseases caused or exacerbated by bacteria, preferably a bacteria of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the disease is a skin disease or a lung disease, more preferably wherein the skin disease is atopic dermatitis, Netherton syndrome, or psoriasis.

In one aspect, the present invention provides a pharmaceutical composition comprising at least one compound according to Formula (I), or a pharmaceutically acceptable salt, tautomer, solvate or hydrate thereof, and a pharmaceutically acceptable excipient.

In one aspect, the present invention provides a compound according to Formula (I) or a pharmaceutically acceptable salt, tautomer, solvate or hydrate thereof, or a pharmaceutical composition comprising a compound of Formula (I), for use as a medicament.

DESCRIPTION OF THE FIGURES

FIG 1 shows quorum sensing signaling in staphylococci to control virulence factor production. The precursor peptide AgrD is processed by AgrB and the mature auto-inducing peptide (AIP) is secreted across the bacterial membrane. AIP binds to the histidine-kinase AgrC of its own cell or on other bacterial cells. AgrC then activates response regulator AgrA by means of phosphorylation. Phospho-AgrA binds to and activates transcription from the agr P2 and P3 promoters, as well as the promoters of the psm operons (RNAIII-independent AgrA regulation). The classic targets of AgrA are under RNAIII-dependent control, comprising several toxins and proteases that are up-regulated and several surface-binding proteins such as protein A, that are down-regulated. Small molecule inhibitors that bind to the DNA-binding domain of AgrA prevent the binding to P2 and P3 promoter, thus blocking continuous production of AIP (P2 driven) and virulence factors (P3 driven) but also block expression of RNAIII-independent AgrA regulation such as production of PSMs. DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to compounds and pharmaceutical compositions comprising the same that can inhibit the transcription of genes under the control of the P3 promoter (e.g., in bacteria, preferably in S. aureus).

Definitions

Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. The herein described and disclosed embodiments, preferred embodiments and very preferred embodiments should apply to all aspects and other embodiments, preferred embodiments and very preferred embodiments irrespective of whether is specifically again referred to or its repetition is avoided for the sake of conciseness.

The articles “a” and “an” are used in this disclosure to refer to one or more than one (i.e., to at least one) of the grammatical object of the article unless the context clearly dictates otherwise. By way of example, “an element” means one element or more than one element.

The term “and/or” is used in this disclosure to mean either “and” or “or” unless indicated otherwise.

The term “optionally substituted” is understood to mean that a given chemical moiety (e.g. an alkyl group) can (but is not required to) be bonded other substituents (e.g. heteroatoms). For instance, an alkyl group that is optionally substituted can be a fully saturated alkyl chain (i.e. a pure hydrocarbon). Alternatively, the same optionally substituted alkyl group can have substituents different from hydrogen. For instance, it can, at any point along the chain be bounded to a halogen atom, a hydroxyl group, or any other substituent described herein. Thus, the term “optionally substituted” means that a given chemical moiety has the potential to contain other functional groups, but does not necessarily have any further functional groups.

The term “alkyl” refers to a straight or branched chain saturated hydrocarbon. Ci-Ce alkyl groups contain 1 to 6 carbon atoms. Examples of a-Ci-Ce alkyl group include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, isopropyl, isobutyl, sec-butyl and tert-butyl, isopentyl and neopentyl.

The terms “alkylene” or “alkylenyl,” as used herein, refer to a straight or branched hydrocarbon chain bi-radical derived from alkyl, as defined herein, wherein one hydrogen of said alkyl is cleaved off generating the second radical of said alkylene. Examples of alkylene are, by way of illustration, -CH ₂ -, -CH2-CH2-, -CH(CH ₃ )-, -CH2-CH2-CH2-, -CH(CH ₃ )-CH ₂ -, or -CH(CH ₂ CH ₃ )-.

The term “cycloalkyl” means monocyclic saturated carbon ring containing 3-7 carbon atoms. Examples of cycloalkyl groups include, without limitations, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

The term “aryl” refers to cyclic, aromatic hydrocarbon groups that have 1 to 2 aromatic rings, including monocyclic or bicyclic groups such as phenyl and naphthyl. A Ce-Cio aryl group contains between 6 and 10 carbon atoms; preferably 6 or 10 carbon atoms. When containing two aromatic rings (bicyclic, etc.), the aromatic rings of the aryl group may be fused (e.g., naphthyl). The aryl group may be optionally substituted by one or more substituents, e.g., 1 to 5 substituents, at any point of attachment. Exemplary substituents include, but are not limited to, -H, -halogen, -O-Ci-Ce alkyl, -Ci-Ce alkyl, -OH, -NH ₂ , -NH(Ci-Ce alkyl), and - N(Ci-Ce alkyl) ₂ ,. The substituents (e.g., alkyl groups) can themselves be optionally substituted.

Unless otherwise specifically defined, “heteroaryl” means a monovalent monocyclic or bicyclic aromatic radical of 5 to 10 ring atoms containing one or more ring heteroatoms selected from N, S, P, and O, the remaining ring atoms being C. Preferably the heteroatom is selected from N, S, and O, more preferably N and O. The aromatic radical is optionally substituted independently with one or more substituents described herein. Examples include, but are not limited to, furyl, thienyl, pyrrolyl, pyridyl, pyrazolyl, pyrimidinyl, imidazolyl, isoxazolyl, oxazolyl, oxadiazolyl, and pyrazinyl.

The terms “heterocyclyl” or “heterocycloalkyl” or “heterocycle” refer to monocyclic or bicyclic saturated or partially saturated 4- to 11 -membered rings containing carbon and heteroatoms taken from O, N, and S (preferably O and N) and wherein the ring or rings do not comprise delocalized 7i electrons (aromaticity) shared among the ring carbons or heteroatoms. Heterocyclyl rings include, but are not limited to, oxetanyl, azetadinyl, tetrahydrofuranyl, pyrrolidinyl, oxazolinyl, oxazolidinyl, thiazolinyl, thiazolidinyl, pyranyl, thiopyranyl, tetrahydropyranyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, [1,4] diazepane, and [1,2] diazepane. In some embodiments the heterocyclyl ring is a fused bicyclic heterocycle, e.g., as in, without limitation, Compounds 129, 130, and 153, e.g., R ⁸¹ and R ⁸² of Compounds 129, 130, and 153.

In some embodiments, the heterocyclyl or heterocycloalkyl group is a spirocyclic heterocycle. As used herein a spirocyclic heterocycle or spiroheterocycle is understood to mean a bicyclic ring system in which both rings are connected through a single atom, and wherein at least one of the rings is a heterocycle (e.g., at least one of the rings is azetidine, pyrrolidine, morpholine, or piperidine). Exemplary spirocyclic heterocycles are, without limitation, compounds 124, 125, 126, 127, 128, 149, 150, 151, and 152, e.g., R ⁸¹ and R ⁸² of Compounds 124, 125, 126, 127, 128, 149, 150, 151, and 152.

As used herein, the term “halo” or “halogen” means fluoro (F), chloro (Cl), bromo (Br), or iodo (I).

The term “oxo” refers to a carbonyl functional group composing a carbon atom doublebonded to an oxygen atom. It can be abbreviated herein as “oxo”, as C(O), or as C=O.

The invention also includes pharmaceutical compositions comprising an effective amount of a disclosed compound and a pharmaceutically acceptable carrier. The terms “pharmaceutically acceptable” or “therapeutically acceptable” refers to a substance which does not interfere with the effectiveness or the biological activity of the active ingredients and which is not toxic to the host. Representative “pharmaceutically acceptable salts” include, e.g., water- soluble and water-insoluble salts, such as the acetate, amsonate (4,4-diaminostilbene-2,2- disulfonate), benzenesulfonate, benzonate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium, calcium edetate, camsylate, carbonate, chloride, citrate, clavulariate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, hydroiodide, sethionate, lactate, lactobionate, laurate, magnesium, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate (l,l-methene-bis-2-hydroxy-3- naphthoate, einbonate), pantothenate, phosphate/diphosphate, picrate, poly galacturonate, propionate, p-toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, teoclate, tosylate, triethiodide, and valerate salts.

The term “tautomers” refers to a set of compounds that have the same number and type of atoms, but differ in bond connectivity and are in equilibrium with one another. A “tautomer” is a single member of this set of compounds. Typically a single tautomer is drawn but it is understood that this single structure is meant to represent all possible tautomers that might exist. Examples include enol-ketone tautomerism. When a ketone is drawn it is understood that both the enol and ketone forms are part of the present disclosure.

The term “solvate” refers to a complex of variable stoichiometry formed by a solute and solvent. Such solvents for the purpose of the invention may not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, MeOH, EtOH, and AcOH. Solvates wherein water is the solvent molecule are typically referred to as “hydrates.” Hydrates include compositions containing stoichiometric amounts of water, as well as compositions containing variable amounts of water.

The term “carrier”, as used in this disclosure, encompasses carriers, excipients, and diluents and means a material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting a pharmaceutical agent from one organ, or portion of the body, to another organ, or portion of the body of a subject.

The term “disorder” is used in this disclosure to mean, and is used interchangeably with, the terms disease, condition, or illness, unless otherwise indicated.

The term “administer”, “administering”, or “administration” as used in this disclosure refers to either directly administering a disclosed compound or pharmaceutically acceptable salt of the disclosed compound or a composition to a subject, or administering a prodrug derivative or analog of the compound or pharmaceutically acceptable salt of the compound or composition to the subject, which can form an equivalent amount of active compound within the subject's body.

The term “ortholog”, as used herein, denotes the well-known meaning of this term. In this art, orthologs are genes in different species which evolved from a common ancestral gene. Due to their separation following a speciation event, orthologs may diverge, but usually have similarity at the sequence and structure levels; furthermore, orthologs usually have identical functions. Orthology is a type of homology. In this application, the term ortholog is used to include the ortholog gene (DNA or RNA) or the peptide/protein product of the ortholog. Sometimes the peptide/protein product of the ortholog is referred to as “ortholog product' or simply "ortholog”. The meaning is evident from the context (e.g., an anti-virulence compositions of the present invention may include an anti- virulence agent capable of reducing the virulence of bacterium that expresses peptides or proteins that may be referred to as orthologs of AgrA - that is, products of an ortholog gene of Staphylococcus aureus AgrA from another bacterium, such as Streptococcus pyogenes). In certain aspects, an ortholog of AgrA produces proteins/peptides that share greater than about 70%, about 80%, or about 90% identity with the amino acid sequence of the gene product of AgrA.

The terms “reducing” and “inhibiting” have their commonly understood meaning of lessening or decreasing.

The expression “reducing the virulence of bacteria that express AgrA”, as used herein, typically and preferably, refers to inhibiting the synthesis of one or more virulence factors by said bacteria by the inventive compounds of Formula (I) or the inventive compositions, preferably pharmaceutical compositions, comprising an inventive compound of Formula (I). Examples for AgrA-regulated virulence factors are cell-surface associated proteins, such as protein A (SpA) and fibronectin-binding proteins, secreted toxins such as a-hemolysin/a-toxin (Hla), 5-hemolysin (Hid), phenol-soluble-modulins (PSMs), Panton-Valentine leukocidin (PVL), leukotoxin E and D (LukED), leukotoxin G and H (LukGH) or secreted proteases such as SspA or aureolysin. In a preferred example and embodiment of the present invention, said reducing the virulence of bacteria that express AgrA is inhibiting the synthesis of one or more virulence factor selected from PSMa, RNAIII and its downstream targets. In a preferred example and embodiment of the present invention, said reducing the virulence of bacteria that express AgrA is inhibiting the synthesis of PSMa. In a preferred example and embodiment of the present invention, said reducing the virulence of bacteria that express AgrA is inhibiting the synthesis of RNAIII and/or its downstream targets, preferably of RNAIII.

The term “inhibiting the synthesis of one or more virulence factors” as used herein shall refer to a complete or partial inhibition (preferably more than 20%, further preferably more than 30%, further preferably more than 50%, further preferably more than 90%, still more preferably more than 95% or even more than 99%) of the synthesis of one or more virulence factors, as compared to the synthesis of one or more virulence factors by said bacteria in the absence of the inventive compounds of Formula (I) or the inventive compositions, preferably pharmaceutical compositions, comprising an inventive compound of Formula (I) or as compared to an inventive method where no such inventive compounds of Formula (I) or the inventive compositions, preferably pharmaceutical compositions, comprising an inventive compound of Formula (I) are applied or used.

Virulence factors as contemplated herein include any molecules expressed and secreted by bacteria to promote colonization and/or adhesion in a host subject, promote inflammation in the host tissue, promote evasion of the host’s immune response and obtain nutrition from the host subject. Virulence factors can also include both exotoxins and endotoxins. Non-limiting examples of virulence factors inhibited by an inventive compound of Formula (I) or an inventive composition, preferably an inventive pharmaceutical composition, comprising an inventive compound of Formula (I), as described herein, include one or more of toxins (e.g., a, P, y, y-variant, and 8-hemolysins, PSMs (e.g., PSMa), Panton-Valentine leukocidin (PVL), leukotoxin E and D (LukED), leukotoxin G and H (LukGH), enterotoxins (e.g., enterotoxin B), exfoliative toxin), proteases (e.g., serine proteases, metalloproteases and cysteine proteases), nuclease, lipase, coagulase, hyaluronidase, fibronectin-binding protein, clumping factor, pyrogenic toxin superantigen (e.g., TSST-1). In a preferred embodiment, the virulence factor inhibited is RNAIII and/or its downstream targets or PSMa.

Depending on the severity of the infection or if used in a preventive manner, the antivirulence drugs can be administered alone or in combination with therapeutic agents such as antibiotics typically and preferably used for prevention and treatment of infections caused by bacteria such as Staphylococcus, primarily by Staphylococcus aureus (e.g., pneumonia, blood stream infections, acute skin and skin structure infections).

The anti- virulence drugs can be administered alone or in combination with therapeutic agents typically and preferably used for prevention and treatment of chronic inflammatory skin disease (e.g., atopic dermatitis, Netherton syndrome, psoriasis) which are exacerbated by bacteria such as Staphylococcus, primarily by Staphylococcus aureus.

The term “antibiotic”, as used herein, refers to an antimicrobial agent or anti-infective that kills bacteria (bactericidal antibiotics) or inhibits growth and/or metabolism of bacteria (bacteriostatic antibiotics). Antibiotics are well-known to the skilled person in the art, and specific and preferred examples thereof include penicillins, cephalosporins, polymyxins, rifamycins, lipiarmycins, quinolones, sulfonamides, macrolides, oxazolidinones, lincosamides and tetracyclines.

The term "treating", “treatment” or "therapy" as used herein refers to means of obtaining a desired physiological effect. The effect may be therapeutic in terms of partially or completely curing a disease or a condition and/or symptoms attributed to the disease or the condition. The term refers to inhibiting the disease or condition, i.e. arresting its development; or ameliorating the disease or condition, i.e. causing regression of the disease or condition.

The term "prevention" as used herein refers to means of preventing or delaying the onset of disease or condition and/or symptoms attributed to the disease or condition.

As used herein, the terms "subject" or "animal" or "patient" or "mammal," refers to any subject, particularly a mammalian subject, for whom diagnosis, prognosis, prophylaxis or therapy is desired, for example, a human or a domesticated mammal such as a dog, cat or horse or a food animal such as a cow or sheep or pig, preferably to a human. Thus, in a preferred embodiment of the present invention, said subject is a human.

As used herein, the term "for use" as used in "composition for use in treatment or prevention of a disease” shall disclose also the corresponding method of treatment or prevention and the corresponding use of a preparation for the manufacture of a medicament for the treatment or prevention of a disease".

A "therapeutically effective amount" or "effective amount" is the amount of a compound or pharmaceutical composition in accordance with the present invention that will elicit the biological or medical response of a subject, preferably a human subject that is being sought by the researcher, veterinarian, medical doctor or other clinician. The term “therapeutic administration”, as used herein, should refer to the administration of therapeutically effective amount. In particular, the terms “effective”, “effective amount”, and “therapeutically effective amount”, as used herein, typically and preferably, refer to that amount of an inventive compounds of Formula (I) or an inventive composition, preferably an inventive pharmaceutical composition, comprising an inventive compound of Formula (I) that reduces the virulence of a bacterium or that results in amelioration of symptoms or a prolongation of survival in a subject with a bacteria related disease or disorder. The term "effective amount" is used generically herein to refer to the amount of a given compound or in case of a mixture the combined amount of mixture components that provides a measurable effect for a listed function. It will be understood by one of ordinary skill in the art, that for a given application, the effective amount can be determined by application of routine experimentation and without undue experimentation by methods that are described herein or that are known in the art. The term "therapeutically effective amount" is used generically herein to refer to the amount of a given compound or in case of a mixture the combined amount of mixture components when administered to the individual including a human or non-human animal, that provides a measurable therapeutic effect for a listed disease, disorder or condition to at least partially ameliorate a symptom of such disease, disorder or condition. The result of treatment can be partially or completely alleviating, inhibiting, preventing, ameliorating and/or relieving the disorder, condition or one or more symptoms thereof.

Compounds of the Invention

In one aspect, the present invention provides a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein:

R ¹ is independently selected from -H, halogen, -Ci-Ce alkyl, and -Ci-Ce alkoxy, wherein said alkyl and alkoxy is each optionally substituted with one or more R ¹¹ ;

R ³ is independently selected from -H, halogen, Ci-Ce alkyl, -Ci-Ce alkoxy, and -C3-C6 cycloalkyl, wherein said alkyl, alkoxy, and cycloalkyl is each optionally substituted by one or more R ¹¹ ;

R ^8A and R ^8B are each independently, at each occurrence, -H or -CH3;

R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C3-C8 cycloalkyl, -Ci-Ce alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, - Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkylene-(C6-Cio aryl), 5- to 10-membered heteroaryl, and -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, - (OCH ₂ -CH ₂ )I- ₂ -OH, -CI-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); and wherein said cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is each independently optionally substituted with one or more R ⁸⁵ ;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl;

R ⁸⁵ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, -OH, - N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, -Ce-Cio aryl, and oxo; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more R ⁸⁶ ;

R ⁸⁶ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, -OH, - Ci-Ce alkylene-OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl, and oxo;

R ¹¹ is independently, at each occurrence, halogen or -OH; m is independently 1 or 2; and n is independently 0 or 1.

In some embodiments, the compound is a compound of Formula (I- A), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ^8A , R ^8B , R ⁸¹ , R ⁸² , m and n are as defined in Formula (I). In some embodiments, the compound is a compound of Formula (I-Al), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ⁸¹ , R ⁸² , and m are as defined in Formula (I). In some embodiments, the compound is a compound of Formula (I-A2), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ⁸¹ , R ⁸² and m are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-A3), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ⁸¹ , R ⁸² , and m are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-A4), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ⁸¹ , R ⁸² , and m are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-B), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ³ , R ^8A , R ^8B , R ⁸¹ , R ⁸² , m and n are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-Bl), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ³ , R ⁸¹ , R ⁸² , and m are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-B2), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof:

wherein R ³ , R ⁸¹ , R ⁸² and m are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-B3), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ³ , R ⁸¹ , R ⁸² , and m are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-B4), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ³ , R ⁸¹ , R ⁸² , and m are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-C), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof:

wherein R ^8A , R ^8B , R ⁸¹ , R ⁸² , and n are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-Cl), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ⁸¹ and R ⁸² are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-C2), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ⁸¹ and R ⁸² are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-C3), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof:

wherein R ⁸¹ and R ⁸² are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-C4), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ⁸¹ and R ⁸² are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-D), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ^8A , R ^8B , R ⁸¹ , R ⁸² , and n are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-Dl), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof:

wherein R ⁸¹ and R ⁸² are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-D2), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ⁸¹ and R ⁸² are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-D3), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ⁸¹ and R ⁸² are as defined in Formula (I).

In some embodiments, the compound is a compound of Formula (I-D4), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein R ⁸¹ and R ⁸² are as defined in Formula (I).

In some embodiments, R ¹ is independently selected from -H, halogen, -C1-C4 alkyl, and -C1-C4 alkoxy, wherein said alkyl and alkoxy is each optionally substituted with one or more R ¹¹ .

In some embodiments, R ¹ is independently selected from -H, -F, -Cl, -C1-C4 alkyl, and -C1-C4 alkoxy, wherein said alkyl and alkoxy is each optionally substituted with one or more F, Cl, or -OH.

In some embodiments, R ¹ is independently selected from -H, -F, -C1-C2 alkyl, and -Ci- C2 alkoxy, wherein said alkyl and alkoxy is each optionally substituted with one or more -F or -OH.

In some embodiments, R ¹ is independently selected from -H, -F, and -C1-C2 alkyl, wherein said alkyl is optionally substituted with one or more -F or -OH.

In some embodiments, R ¹ is independently selected from -H, -F, and -C1-C2 alkyl.

In some embodiments, R ¹ is independently selected from -H and -C1-C2 alkyl.

In preferred embodiments, R ¹ is -CH3.

In some embodiments, R ³ is independently selected from -H, halogen, -C1-C4 alkyl, - C1-C4 alkoxy, and -C3-C6 cycloalkyl, wherein said alkyl, alkoxy, and cycloalkyl is each optionally substituted by one or more R ¹¹ .

In some embodiments, R ³ is independently selected from -H, halogen, -C1-C3 alkyl, - C1-C3 alkoxy, and -C3-C6 cycloalkyl, wherein said alkyl, alkoxy, and cycloalkyl is each optionally substituted by one or more R ¹¹ .

In some embodiments, R ³ is independently selected from -H, halogen, -C1-C3 alkyl, - C1-C2 alkoxy, and cyclopropyl, wherein said alkyl, alkoxy, and cyclopropyl is each optionally substituted by one or more R ¹¹ .

In some embodiments, R ³ is independently selected from -H, -Cl, -C1-C3 alkyl, -C1-C2 alkoxy, and cyclopropyl, wherein said alkyl is optionally substituted by one or more -F, -Cl, or -OH. In some embodiments, R ³ is independently selected from -H, -Cl, -C1-C3 alkyl, -C1-C2 alkoxy, and cyclopropyl, wherein said alkyl is optionally substituted by one or more -F, preferably wherein said alkyl is unsubstituted or substituted by three -F.

In some embodiments, R ³ is independently selected from -Cl, -C1-C3 alkyl, -C1-C2 alkoxy, and cyclopropyl, wherein said alkyl is optionally substituted by one or more -F, preferably wherein said alkyl is unsubstituted or substituted by three -F.

In some embodiments, R ³ is independently selected from -CH3, -CF3, -Cl, -CH(CH3)2, -OCH3, and cyclopropyl.

In some embodiments, R ³ is -CF3.

In some embodiments, R ³ is -Cl.

In some embodiments, R ³ is -CH(CH3)2.

In some embodiments, R ³ is -OCH3.

In some embodiments, R ³ is cyclopropyl.

In preferred embodiments, R ³ is -CH3. In preferred embodiments, R ¹ and R ³ are both - CH ₃ .

In preferred embodiments, R ^8A and R ^8B are both -H.

In some embodiments, R ^8A and R ^8B are both -H or both -CH3.

In some embodiments, R ^8A and R ^8B are both -CH3.

In some embodiments, R ^8A is -H and R ^8B is -CH3.

In preferred embodiments, n is 0 and m is 1.

In some embodiments, n is 0.

In some embodiments, n is 1.

In some embodiments, m is 1.

In some embodiments, m is 2.

In some embodiments, n is 0 and m is 2.

In some embodiments, n is 1 and m is 1

In some embodiments, n is 1 and m is 2.

In preferred embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl, wherein said alkyl is optionally substituted with one or more -C ₂ -C ₆ alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are both -H.

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl, wherein said alkyl is optionally substituted with one or more -C2-C4 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -Ci-C ₄ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₄ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl, wherein said alkyl is optionally substituted with one or more -C ₂ -C ₄ alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -Ci-C ₄ alkoxy, -NH ₂ , -NH(CI-C ₄ alkyl), -N(CI-C ₄ alkyl) ₂ , or -S(Ci-C ₄ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl, wherein said alkyl is optionally substituted with one or more -C ₂ alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three halogen; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I. ₂ -OH, -Ci-C ₂ alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F, -Cl, or -Br; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), - N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I. ₂ -OH, -Ci-C ₂ alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH ₂ , or -S(Ci-C ₂ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I. ₂ -OH, -OCH ₃ , -NH ₂ , or SCH ₃ .

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -OCH ₃ , or -NH ₂ .

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -OCH3, or -NH ₂ .

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three halogen; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F, -Cl, or -Br; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I. ₂ -OH, -Ci-C ₂ alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH ₂ , or -S(Ci-C ₂ alkyl).

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ -CH ₂ )I. ₂ -OH, -OCH3, -NH ₂ , or -SCH3.

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -OCH3, or -NH ₂ .

In some embodiments, R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -OH, -OCH ₃ , or -NH ₂ .

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl, wherein said alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH2- CH ₂ )I- ₂ -OH, -CI-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl, wherein said alkyl is optionally substituted with one or more -C2-C4 alkynyl, halogen, -OH, -(OCH ₂ - CH ₂ )I- ₂ -OH, -C1-C4 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₄ alkyl); and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl, wherein said alkyl is optionally substituted with one or more -C2-C4 alkynyl, halogen, -OH, -(OCH ₂ - CH ₂ )I- ₂ -OH, -C1-C4 alkoxy, -NH ₂ , -NH(CI-C4 alkyl), -N(CI-C4 alkyl) ₂ , or -S(Ci-C4 alkyl); and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl, wherein said alkyl is optionally substituted with one or more -C ₂ alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- 2-OH, -C1-C2 alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl); and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three halogen; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ - CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl); and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F, -Cl, or -Br; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, - (OCH ₂ -CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl); and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C ₂ alkynyl, -OH, -(OCH ₂ - CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl); and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C2 alkynyl, -OH, -(OCH2- CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH2, or -S(Ci-C2 alkyl); and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -C2 alkynyl, -OH, -(OCH2- CH ₂ )I- ₂ -OH, -OCH3, -NH ₂ , or -SCH ₃ ; and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -OH, -(OCH2-CH2)i-2-OH, -OCH3, or -NH ₂ ; and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; and wherein said alkyl is further optionally substituted with one or two, preferably with exactly one -OH, -OCH3, or -NH2; and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three halogen; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -C2 alkynyl, -OH, -(OCH2-CH2)i-2- OH, -C1-C2 alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl); and R ⁸² is - H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F, -Cl, or -Br; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -C2 alkynyl, -OH, -(OCH2- CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -NH2, -NH(CI-C2 alkyl), -N(CI-C2 alkyl)2, or -S(Ci-C2 alkyl); and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -C2 alkynyl, -OH, -(OCH2-CH2)i-2- OH, -C1-C2 alkoxy, -NH ₂ , -NH(CI-C ₂ alkyl), -N(CI-C ₂ alkyl) ₂ , or -S(Ci-C ₂ alkyl); and R ⁸² is - H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -C2 alkynyl, -OH, -(OCH2-CH2)i-2- OH, -C1-C2 alkoxy, -NH2, or -S(Ci-C2 alkyl); and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -C2 alkynyl, -OH, -(OCH2-CH2)I-2- OH, -OCH3, -NH ₂ , or -SCH ₃ ; and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -OH, -(OCH2-CH2)I-2-OH, -OCH3, or -NH2; and R ⁸² is -H.

In some embodiments, R ⁸¹ is independently selected from -H and -Ci-Ce alkyl; wherein said alkyl is optionally substituted with one to three -F; or wherein said alkyl is optionally substituted with one or two, preferably with exactly one -OH, -OH, -OCH3, or -NH2; and R ⁸² is -H.

In some embodiments, R ⁸¹ and R ⁸² are each -Ci-Ce alkyl.

In some embodiments, R ⁸¹ and R ⁸² are each -C1-C4 alkyl.

In some embodiments, R ⁸¹ and R ⁸² are each -C1-C2 alkyl, preferably wherein R ⁸¹ is - CH ₃ and R ⁸² is -C1-C2 alkyl.

In preferred embodiments, R ⁸¹ is selected from -C3-C8 cycloalkyl and -Ci-Ce alkylene- (C3-C8 cycloalkyl).

In some embodiments, R ⁸¹ is selected from -C3-C8 cycloalkyl and -Ci-Ce alkylene-(C3- Cs cycloalkyl); and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C8 cycloalkyl and -Ci-Ce alkylene-(C3- Cs cycloalkyl), wherein said cycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, - OH, -Ci-Ce alkoxy, -N(R ⁸³ )(R ⁸⁴ ), -Ce-Cio aryl, and oxo; and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C8 cycloalkyl and -C1-C4 alkylene-(C3- Cs cycloalkyl), wherein said cycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, - OH, -C1-C4 alkoxy, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl)2, -Ce-Cio aryl, and oxo; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C8 cycloalkyl and -C1-C4 alkylene-(C3- Cs cycloalkyl), wherein said cycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, - OH, -C1-C4 alkoxy, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl)2, and -Ce-Cio aryl; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H. In some embodiments, R ⁸¹ is selected from -Cs-Cs cycloalkyl and -C1-C3 alkylene-(C3- Cs cycloalkyl), wherein said cycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C3 alkyl, halogen, - OH, -C1-C3 alkoxy, -NH2, -NH(CI-C3 alkyl), -N(CI-C3 alkyl)2, and phenyl; and R ⁸² is -H or - C1-C3 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C7 cycloalkyl and -C1-C3 alkylene-(C3- C7 cycloalkyl), preferably -C3-C6 cycloalkyl and -C1-C3 alkylene-(C3-C6 cycloalkyl), wherein said cycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C3 alkyl, halogen, -OH, -C1-C3 alkoxy, - NH2, -NH(CI-C3 alkyl), -N(CI-C3 alkyl)2, and phenyl; and R ⁸² is -H or -C1-C3 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C7 cycloalkyl and -C1-C3 alkylene-(C3- C7 cycloalkyl), preferably -C3-C6 cycloalkyl and -C1-C3 alkylene-(C3-C6 cycloalkyl), wherein said cycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C3 alkyl, -OH, -NH2, -NH(CI-C3 alkyl), - N(Ci-Cs alkyl)2, and phenyl; and R ⁸² is -H or -C1-C3 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C7 cycloalkyl and -C1-C3 alkylene-(C3- C7 cycloalkyl), preferably -C3-C6 cycloalkyl and -C1-C3 alkylene-(C3-C6 cycloalkyl), wherein said cycloalkyl is each independently substituted with one or two, preferably with exactly one R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -OH, -NH2, -NH(Ci- C3 alkyl), -N(CI-C3 alkyl)2, and phenyl; and R ⁸² is -H or -C1-C3 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C7 cycloalkyl and -C1-C3 alkylene-(C3- C7 cycloalkyl), preferably -C3-C6 cycloalkyl and -C1-C3 alkylene-(C3-C6 cycloalkyl), wherein said cycloalkyl is each independently substituted with one or two, preferably with exactly one R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -OH, -NH2, -NH(Ci- C2 alkyl), -N(CI-C2 alkyl)2, and phenyl; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C7 cycloalkyl and -C1-C3 alkylene-(C3- C7 cycloalkyl), preferably -C3-C6 cycloalkyl and -C1-C3 alkylene-(C3-Ce cycloalkyl), wherein said cycloalkyl is each independently substituted with one or two, preferably with exactly one R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -OH, -N(CI-C2 alky 1)2. and phenyl; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C8 cycloalkyl and -Ci-Ce alkylene-(C3- Cs cycloalkyl), wherein when R ⁸¹ is -C3-C8 cycloalkyl, R ⁸¹ is independently substituted with one or more R ⁸⁵ , and wherein when R ⁸¹ is -Ci-Ce alkylene-(C3-C8 cycloalkyl), R ⁸¹ is unsubstituted; R ⁸⁵ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, - OH, -Ci-Ce alkoxy, -N(R ⁸³ )(R ⁸⁴ ), -Ce-Cio aryl, and oxo; and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -Cs-Cs cycloalkyl and -C1-C4 alkylene-(C3- Cs cycloalkyl), wherein when R ⁸¹ is -Cs-Cs cycloalkyl, R ⁸¹ is independently substituted with one or more R ⁸⁵ , and wherein when R ⁸¹ is -C1-C4 alkylene-(C3-Cs cycloalkyl), R ⁸¹ is unsubstituted; R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, - OH, -C1-C4 alkoxy, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl)2, -Ce-Cio aryl, and oxo; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -Cs-Cs cycloalkyl and -C1-C4 alkylene-(C3- Cs cycloalkyl), wherein when R ⁸¹ is -Cs-Cs cycloalkyl, R ⁸¹ is independently substituted with one or more R ⁸⁵ , and wherein when R ⁸¹ is -C1-C4 alkylene-(C3-Cs cycloalkyl), R ⁸¹ is unsubstituted; R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, - OH, -C1-C4 alkoxy, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl)2, and -Ce-Cio aryl; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -Cs-Cs cycloalkyl and -C1-C3 alkylene-(C3- Cs cycloalkyl), wherein when R ⁸¹ is -C's-C's cycloalkyl, R ⁸¹ is independently substituted with one or more R ⁸⁵ , and wherein when R ⁸¹ is -C1-C3 alkylene-(C3-Cs cycloalkyl), R ⁸¹ is unsubstituted; R ⁸⁵ is independently, at each occurrence, selected from -C1-C3 alkyl, halogen, - OH, -C1-C3 alkoxy, -NH2, -NH(CI-C3 alkyl), -N(CI-C3 alkyl)2, and phenyl; and R ⁸² is -H or - C1-C3 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C7 cycloalkyl and -C1-C3 alkylene-(C3- C7 cycloalkyl), preferably -C3-C6 cycloalkyl and -C1-C3 alkylene-(C3-C6 cycloalkyl), wherein when R ⁸¹ is -C3-C7 cycloalkyl or C3-C6 cycloalkyl, R ⁸¹ is independently substituted with one or more R ⁸⁵ , and wherein when R ⁸¹ is -C1-C3 alkylene-(C3-C7 cycloalkyl) or -C1-C3 alkylene-(C3- Ce cycloalkyl), R ⁸¹ is unsubstituted; R ⁸⁵ is independently, at each occurrence, selected from - C1-C3 alkyl, halogen, -OH, -C1-C3 alkoxy, -NH2, -NH(CI-C3 alkyl), -N(CI-C3 alkyl)2, and phenyl; and R ⁸² is -H or -C1-C3 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C7 cycloalkyl and -C1-C3 alkylene-(C3- C7 cycloalkyl), preferably -C3-C6 cycloalkyl and -C1-C3 alkylene-(C3-C6 cycloalkyl), wherein when R ⁸¹ is -C3-C7 cycloalkyl or C3-C6 cycloalkyl, R ⁸¹ is independently substituted with one or more R ⁸⁵ , and wherein when R ⁸¹ is -C1-C3 alkylene-(C3-C7 cycloalkyl) or -C1-C3 alkylene-(C3- Ce cycloalkyl), R ⁸¹ is unsubstituted; R ⁸⁵ is independently, at each occurrence, selected from - C1-C3 alkyl, -OH, -NH2, -NH(CI-C3 alkyl), -N(CI-C3 alkyl)2, and phenyl; and R ⁸² is -H or -Ci- C alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C7 cycloalkyl and -C1-C3 alkylene-(C3- C7 cycloalkyl), preferably -C3-C6 cycloalkyl and -C1-C3 alkylene-(C3-Ce cycloalkyl), wherein when R ⁸¹ is -C3-C7 cycloalkyl or C3-C6 cycloalkyl, R ⁸¹ is independently substituted with one two, preferably with exactly one R ⁸⁵ , and wherein when R ⁸¹ is -C1-C3 alkylene-(C3-C7 cycloalkyl) or -C1-C3 alkylene-(C3-Ce cycloalkyl), R ⁸¹ is unsubstituted; R ⁸⁵ is independently, at each occurrence, selected from -OH, -NH2, -NH(CI-C3 alkyl), -N(CI-C3 alkyl)2, and phenyl; and R ⁸² is -H or -C1-C3 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C7 cycloalkyl and -C1-C3 alkylene-(C3- C7 cycloalkyl), preferably -C3-C6 cycloalkyl and -C1-C3 alkylene-(C3-Ce cycloalkyl), wherein when R ⁸¹ is -C3-C7 cycloalkyl or C3-C6 cycloalkyl, R ⁸¹ is independently substituted with one two, preferably with exactly one R ⁸⁵ , and wherein when R ⁸¹ is -C1-C3 alkylene-(C3-C7 cycloalkyl) or -C1-C3 alkylene-(C3-Ce cycloalkyl), R ⁸¹ is unsubstituted; R ⁸⁵ is independently, at each occurrence, selected from -OH, -NH2, -NH(CI-C2 alkyl), -N(CI-C2 alkyl)2, and phenyl; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -C3-C7 cycloalkyl and -C1-C3 alkylene-(C3- C7 cycloalkyl), preferably -C3-C6 cycloalkyl and -C1-C3 alkylene-(C3-C6 cycloalkyl), wherein when R ⁸¹ is -C3-C7 cycloalkyl or C3-C6 cycloalkyl, R ⁸¹ is independently substituted with one two, preferably with exactly one R ⁸⁵ , and wherein when R ⁸¹ is -C1-C3 alkylene-(C3-C7 cycloalkyl) or -C1-C3 alkylene-(C3-C6 cycloalkyl), R ⁸¹ is unsubstituted; R ⁸⁵ is independently, at each occurrence, selected from -OH, -N(CI-C2 alkyl)2, and phenyl; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In preferred embodiments, R ⁸¹ is selected from 4- to 8-membered heterocycloalkyl and -Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl).

In some embodiments, R ⁸¹ is selected from 4- to 8-membered heterocycloalkyl and -Ci- Ce alkylene-(4- to 8-membered heterocycloalkyl); and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 4- to 8-membered heterocycloalkyl and -Ci- Ce alkylene-(4- to 8-membered heterocycloalkyl); wherein said heterocycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, -OH, -Ci-Ce alkoxy, -N(R ⁸³ )(R ⁸⁴ ), -Ce-Cio aryl, and oxo; and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 4- to 8-membered heterocycloalkyl and -Ci- C4 alkylene-(4- to 8-membered heterocycloalkyl); wherein said heterocycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C4 alkoxy, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl)2, -Ce-Cio aryl, and oxo; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 4- to 8-membered heterocycloalkyl and -Ci- C4 alkylene-(4- to 8-membered heterocycloalkyl); wherein said heterocycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C4 alkoxy, -Ce-Cio aryl, and oxo; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 4- to 8-membered heterocycloalkyl and -Ci- C4 alkylene-(4- to 8-membered heterocycloalkyl); wherein said heterocycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C4 alkoxy, and oxo; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 4- to 8-membered heterocycloalkyl and -Ci- C4 alkylene-(4- to 8-membered heterocycloalkyl); wherein said heterocycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, and oxo; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 4- to 8-membered heterocycloalkyl and -Ci- C3 alkylene-(4- to 8-membered heterocycloalkyl); wherein said heterocycloalkyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C2 alkyl, halogen, -OH, and oxo; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 4- to 8-membered heterocycloalkyl and -Ci- C3 alkylene-(4- to 8-membered heterocycloalkyl); wherein said heterocycloalkyl each independently comprises one or more heteroatoms selected from N and O, preferably one or two heteroatoms selected from N and O, and wherein said heterocycloalkyl is each independently substituted with one or more R ⁸⁵ ; wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C2 alkyl, halogen, -OH, and oxo; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 4- to 8-membered heterocycloalkyl and -Ci- C3 alkylene-(4- to 8-membered heterocycloalkyl); wherein said heterocycloalkyl each independently comprises one or more heteroatoms selected from N and O, preferably one or two heteroatoms selected from N and O, and wherein said heterocycloalkyl is each independently substituted with one or more R ⁸⁵ ; wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C2 alkyl and oxo; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 4- to 6-membered heterocycloalkyl and -Ci- C3 alkylene-(4- to 6-membered heterocycloalkyl); wherein said heterocycloalkyl is selected from azetidine, pyrrolidine, tetrahydrofuran, piperidine, piperazine, tetrahydropyran, and morpholine; and wherein said heterocycloalkyl is each independently substituted with one or more R ⁸⁵ ; wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C2 alkyl and oxo; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 4- to 6-membered heterocycloalkyl and -Ci- C3 alkylene-(4- to 6-membered heterocycloalkyl); wherein said heterocycloalkyl is selected from azetidine, pyrrolidine, tetrahydrofuran, piperidine, piperazine, tetrahydropyran, and morpholine; wherein when R ⁸¹ is 4- to 6-membered heterocycloalkyl, said heterocycloalkyl is optionally substituted with one or more, preferably one or two -C1-C2 alkyl; and wherein when R ⁸¹ is -C1-C3 alkylene-(4- to 6-membered heterocycloalkyl), said heterocycloalkyl is optionally substituted with one or more, preferably one or two -C1-C2 alkyl or oxo; and R ⁸² is -H or -Ci- C2 alkyl, preferably R ⁸² is -H.

In preferred embodiments, R ⁸¹ is selected from -Ce-Cio aryl and -Ci-Ce alkylene-(C6- C10 aryl).

In some embodiments, R ⁸¹ is selected from -Ce-Cio aryl and -Ci-Ce alkylene-(C6-Cio aryl); and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -Ce-Cio aryl and -Ci-Ce alkylene-(C6-Cio aryl); wherein said aryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, -OH, -Ci-Ce alkoxy, -N(R ⁸³ )(R ⁸⁴ ), -Ce-Cio aryl, and oxo; and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -Ce-Cio aryl and -Ci-Ce alkylene-(C6-Cio aryl); wherein said aryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, -OH, -Ci-Ce alkoxy, -N(R ⁸³ )(R ⁸⁴ ), and -Ce-Cio aryl; and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -Ce-Cio aryl and -C1-C4 alkylene-(C6-Cio aryl); wherein said aryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C4 alkoxy, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl)2, and -Ce-Cio aryl; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H. In some embodiments, R ⁸¹ is selected from -Ce-Cio aryl and -C1-C4 alkylene-(C6-Cio aryl); wherein said aryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C4 alkoxy, -NH2, -NH(CI-C4 alkyl), and -N(CI-C4 alkyl)2; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -Ce-Cio aryl and -C1-C2 alkylene-(C6-Cio aryl); wherein said aryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C2 alkyl, halogen, -OH, -C1-C2 alkoxy, -NH2, -NH(CI-C2 alkyl), and -N(CI-C2 alkyl)2; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from -Ce-Cio aryl and -C1-C2 alkylene-(C6-Cio aryl); wherein said aryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C2 alkyl, halogen, -OH, and -C1-C2 alkoxy; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from phenyl and -C1-C2 alkylene-(phenyl); wherein said phenyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C2 alkyl, halogen, -OH, and -C1-C2 alkoxy; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from phenyl and -C1-C2 alkylene-(phenyl); wherein said phenyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C2 alkyl, -F, -Cl, -OH, and -C1-C2 alkoxy; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from phenyl and -C1-C2 alkylene-(phenyl); wherein said phenyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C2 alkyl, -Cl, -OH, and -C1-C2 alkoxy; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from phenyl and -C1-C2 alkylene-(phenyl); wherein said phenyl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -Cl, -OH, and -C1-C2 alkoxy; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In preferred embodiments, R ⁸¹ is selected from 5- to 10-membered heteroaryl and -Ci- Ce alkyl ene-(5- to 10-membered heteroaryl).

In some embodiments, R ⁸¹ is selected from 5- to 10-membered heteroaryl and -Ci-Ce alkylene-(5- to 10-membered heteroaryl); and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H. In some embodiments, R ⁸¹ is selected from 5- to 10-membered heteroaryl and -Ci-Ce alkyl ene-(5- to 10-membered heteroaryl); wherein said heteroaryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, -OH, -Ci-Ce alkoxy, -N(R ⁸³ )(R ⁸⁴ ), -Ce-Cio aryl, and oxo; and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 5- to 10-membered heteroaryl and -C1-C4 alkyl ene-(5- to 10-membered heteroaryl); wherein said heteroaryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C4 alkoxy, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl), - Ce-Cio aryl, and oxo; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 5- to 10-membered heteroaryl and -C1-C4 alkyl ene-(5- to 10-membered heteroaryl); wherein said heteroaryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, and -OH; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 5- to 6-membered heteroaryl and -C1-C4 alkyl ene-(5- to 6-membered heteroaryl); wherein said heteroaryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, and -OH; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 5- to 6-membered heteroaryl and -C1-C4 alkyl ene-(5- to 6-membered heteroaryl); wherein said heteroaryl each independently comprises one or more heteroatoms selected firomN, O, and S, preferably one or two heteroatoms selected from N, O, and S, and wherein said heteroaryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, and -OH; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 5- to 6-membered heteroaryl and -C1-C4 alkyl ene-(5- to 6-membered heteroaryl); wherein said heteroaryl is each independently selected from pyrrole, furan, thiophene, oxazole, imidazole, pyridine, pyrazole, isoxazole, triazole, and pyrimidine; and wherein said heteroaryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, and -OH; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 5- to 6-membered heteroaryl and -C1-C4 alkyl ene-(5- to 6-membered heteroaryl); wherein said heteroaryl is each independently selected from pyrrole, furan, thiophene, oxazole, imidazole, and pyridine; and wherein said heteroaryl is each independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, and -OH; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 5- to 6-membered heteroaryl and -C1-C4 alkyl ene-(5- to 6-membered heteroaryl); wherein said heteroaryl is each independently selected from pyrrole, furan, thiophene, oxazole, imidazole, and pyridine; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is selected from 5- to 6-membered heteroaryl and -C1-C2 alkyl ene-(5- to 6-membered heteroaryl); wherein said heteroaryl is each independently selected from pyrrole, furan, thiophene, oxazole, imidazole, and pyridine; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is -Ci-Ce alkylene-(5- to 10-membered heteroaryl).

In some embodiments, R ⁸¹ is -Ci-Ce alkylene-(5- to 10-membered heteroaryl); and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said heteroaryl is independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -Ci-Ce alkyl, halogen, -OH, -Ci-Ce alkoxy, - N(R ⁸³ )(R ⁸⁴ ), -Ce-Cio aryl, and oxo; and R ⁸² is -H or -Ci-Ce alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is -C1-C4 alkylene-(5- to 10-membered heteroaryl); wherein said heteroaryl is independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C4 alkoxy, - NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl), -Ce-Cio aryl, and oxo; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is -C1-C4 alkylene-(5- to 10-membered heteroaryl); wherein said heteroaryl is independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, and -OH; and R ⁸² is - H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is -C1-C4 alkylene-(5- to 6-membered heteroaryl); wherein said heteroaryl is independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, and -OH; and R ⁸² is - H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is -C1-C4 alkylene-(5- to 6-membered heteroaryl); wherein said heteroaryl independently comprises one or more heteroatoms selected from N, O, and S, preferably one or two heteroatoms selected from N, O, and S, and wherein said heteroaryl is independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, and -OH; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is -C1-C4 alkylene-(5- to 6-membered heteroaryl); wherein said heteroaryl is independently selected from pyrrole, furan, thiophene, oxazole, imidazole, pyridine, pyrazole, isoxazole, triazole, and pyrimidine; and wherein said heteroaryl is independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, and -OH; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is -C1-C4 alkylene-(5- to 6-membered heteroaryl); wherein said heteroaryl is independently selected from pyrrole, furan, thiophene, oxazole, imidazole, and pyridine; and wherein said heteroaryl is independently substituted with one or more R ⁸⁵ , and wherein R ⁸⁵ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, and -OH; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is -C1-C4 alkylene-(5- to 6-membered heteroaryl); wherein said heteroaryl is independently selected from pyrrole, furan, thiophene, oxazole, imidazole, and pyridine; and R ⁸² is -H or -C1-C4 alkyl, preferably R ⁸² is -H.

In some embodiments, R ⁸¹ is -C1-C2 alkylene-(5- to 6-membered heteroaryl); wherein said heteroaryl is independently selected from pyrrole, furan, thiophene, oxazole, imidazole, and pyridine; and R ⁸² is -H or -C1-C2 alkyl, preferably R ⁸² is -H.

In preferred embodiments, R ⁸² is -H or Ci-Ce alkyl, preferably R ⁸² is -H or C1-C2 alkyl, more preferably R ⁸² is H.

In some embodiments, R ⁸² is -H or -C1-C2 alkyl.

In some embodiments, R ⁸² is -H.

In preferred embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11- membered heterocycloalkyl is optionally substituted with one or more R ⁸⁶ .

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11- membered heterocycloalkyl is optionally substituted with one or more R ⁸⁶ , wherein R ⁸⁶ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C4 alkylene- OH, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl)2, -C1-C4 alkylene-Ndt, -C1-C4 alkylene-NH(Ci- C4 alkyl), -C1-C4 alkylene-N(Ci-C4 alkyl)2, -C3-C6 cycloalkyl, and oxo.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11- membered heterocycloalkyl is optionally substituted with one or more R ⁸⁶ , wherein R ⁸⁶ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C2 alkylene- OH, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl)2, -C1-C2 alkylene-NH2, -C1-C2 alkylene-NH(Ci- C4 alkyl), -C1-C2 alkylene-N(Ci-C4 alkyl)2, -C3-C6 cycloalkyl, and oxo.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said heterocycloalkyl comprises one or more heteroatoms selected from O and N, and wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more R ⁸⁶ , wherein R ⁸⁶ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C2 alkylene-OH, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl)2, -C1-C2 alkylene-NH2, -C1-C2 alkylene-NH(Ci-C4 alkyl), -C1-C2 alkylene-N(Ci-C4 alkyl)2, -C3-C6 cycloalkyl, and oxo.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said heterocycloalkyl comprises one or two heteroatoms selected from O and N, and wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or two R ⁸⁶ , wherein R ⁸⁶ is independently, at each occurrence, selected from -C1-C4 alkyl, halogen, -OH, -C1-C2 alkylene-OH, -NH2, -NH(CI-C4 alkyl), -N(CI-C4 alkyl)2, -C1-C2 alkylene-NH2, -C1-C2 alkylene-NH(Ci-C4 alkyl), -C1-C2 alkylene-N(Ci-C4 alkyl)2, -C3-C6 cycloalkyl, and oxo.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said heterocycloalkyl comprises one or two heteroatoms selected from O and N, and wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or two R ⁸⁶ , wherein R ⁸⁶ is independently, at each occurrence, selected from-Ci-C4 alkyl, -F, -Cl, -OH, -C1-C2 alkylene- OH, -NH2, -N(CI-C2 alkyl)2, -C1-C2 alkylene-NH2, -C3-C6 cycloalkyl, and oxo.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said heterocycloalkyl comprises one or two heteroatoms selected from O and N, and wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or two R ⁸⁶ , wherein R ⁸⁶ is independently, at each occurrence, selected from -C1-C4 alkyl, -F, -OH, -C1-C2 alkylene-OH, -NH2, -N(CI-C2 alkyl)2, -C1-C2 alkylene-NH2, -C3-C6 cycloalkyl (preferably cyclopropyl), and oxo.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said heterocycloalkyl is selected from azetidine, oxetane, pyrrolidine, tetrahydrofuran, piperidine, piperazine, morpholine, azepane, oxazepane (preferably [1,4] oxazepane), diazepane (preferably [1,4] diazepane), 7- to 11 -membered spirocyclic heterocycloalkyl, and 8- to 10- membered fused bicyclic heterocycloalkyl; and wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or two R ⁸⁶ , wherein R ⁸⁶ is independently, at each occurrence, selected from -C1-C4 alkyl, -F, -OH, -C1-C2 alkylene-OH, -NH2, -N(CI-C2 alkyl)2, -C1-C2 alkylene-NH2, -C3-C6 cycloalkyl (preferably cyclopropyl), and oxo.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said heterocycloalkyl is selected from azetidine, pyrrolidine, piperidine, piperazine, morpholine, azepane, oxazepane (preferably [1,4] oxazepane), diazepane (preferably [1,4] diazepane), 7- to 11-membered spirocyclic heterocycloalkyl, and 8- to 10-membered fused bicyclic heterocycloalkyl; and wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or two R ⁸⁶ , wherein R ⁸⁶ is independently, at each occurrence, selected from -C1-C4 alkyl, -F, -OH, -C1-C2 alkylene-OH, -NH2, -N(CI-C2 alkyl)2, -C1-C2 alkylene-NH2, -C3- Ce cycloalkyl (preferably cyclopropyl), and oxo.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a pyrrolidine, piperidine, or morpholine, wherein said pyrrolidine, piperidine, or morpholine is optionally substituted with one or two R ⁸⁶ .

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a pyrrolidine or morpholine, wherein said pyrrolidine or morpholine is optionally substituted with one or two R ⁸⁶ .

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a pyrrolidine, wherein said pyrrolidine is optionally substituted with one or two R ⁸⁶ .

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a pyrrolidine, wherein said pyrrolidine is unsubstituted.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form an N-bound pyrrolidine, wherein said N-bound pyrrolidine is optionally substituted with one or two R ⁸⁶ .

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form an N-bound pyrrolidine, wherein said N-bound pyrrolidine is unsubstituted.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a morpholine, wherein said morpholine is optionally substituted with one or two R ⁸⁶ .

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a morpholine, wherein said morpholine is unsubstituted.

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form an N-bound morpholine, wherein said an N-bound morpholine is optionally substituted with one or two R ⁸⁶ .

In some embodiments, R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form an N-bound morpholine, wherein said an N-bound morpholine is unsubstituted.

In some embodiments, the compound is a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein:

R ¹ is -CH ₃ ;

R ³ is -CH ₃ ;

R ^8A and R ^8B are each independently, at each occurrence, -H or -CH ₃ ;

R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C ₃ -Cs cycloalkyl, -Ci-Ce alkylene-(C ₃ -Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, - Ci-C ₆ alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkylene-Ce-Cio aryl, 5- to 10-membered heteroaryl, or -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); wherein said -C ₃ -Cs cycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), or -Ci-Ce alkoxy; wherein said 5- to 10-membered heteroaryl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl, or oxo;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl; m is independently 1 or 2; and n is independently 0 or 1.

In some embodiments, the compound is a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: wherein:

R ¹ is -CH ₃ ;

R ³ is -CH ₃ ;

R ^8A and R ^8B are each independently, at each occurrence, -H or -CH3;

R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -Cs-Cs cycloalkyl, -Ci-Ce alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, - Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkylene-Ce-Cio aryl, 5- to 10-membered heteroaryl, or -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); wherein said -C3-C8 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more halogen, -OH, or -Ci-Ce alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl, or oxo;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl; m is independently 1 or 2; and n is independently 0 or 1.

In some embodiments, the compound is a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: Formula (I) wherein:

R ¹ is -CH ₃ ;

R ³ is -CH ₃ ;

R ^8A and R ^8B are each independently, at each occurrence, -H or -CH3;

R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, - C1-C3 alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -C1-C3 alkylene-Ce-Cio aryl, 5- to 10-membered heteroaryl, or -C1-C3 alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C ₂ alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I-2-OH, -Ci-C ₂ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₂ alkyl); wherein said -C3-C8 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -C1-C2 alkyl or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more halogen, -OH, or -C1-C2 alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -C1-C2 alkyl, halogen, -OH, -Ci-C ₃ alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -CI-C ₃ alkylene-N(R ⁸³ )(R ⁸⁴ ), -C ₃ -C6 cycloalkyl (preferably cyclopropyl), or oxo;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3; m is independently 1 or 2; and n is independently 0 or 1.

In some embodiments, the compound is a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: Formula (I) wherein:

R ¹ is -CH ₃ ;

R ³ is -CH ₃ ;

R ^8A and R ^8B are each independently, at each occurrence, -H or -CH ₃ ;

R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci-Ce alkyl,

-C ₃ -Cs cycloalkyl, -Ci-C ₃ alkylene-(C ₃ -C6 cycloalkyl), 4- to 6-membered heterocycloalkyl, -

Ci-C ₃ alkylene-(4- to 6-membered heterocycloalkyl), -Ce aryl, -Ci-C ₃ alkylene-Ce aryl, 5- to 6- membered heteroaryl, or -Ci-C ₃ alkylene-(5- to 6-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₂ alkyl); wherein said -C ₃ -Ce cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 6-membered heterocycloalkyl is each independently optionally substituted with one or more -C1-C2 alkyl or oxo; wherein said -Ce aryl is each independently optionally substituted with one or more halogen, -OH, or -C1-C2 alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -C1-C2 alkyl, halogen, -OH, -Ci-C ₃ alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -CI-C ₃ alkylene-N(R ⁸³ )(R ⁸⁴ ), -C ₃ -Ce cycloalkyl (preferably cyclopropyl), or oxo;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH ₃ ; m is independently 1 or 2; and n is independently 0 or 1.

In some embodiments, the compound is a compound of Formula (I), or a pharmaceutically acceptable salt, hydrate, or tautomer thereof: Formula (I) wherein:

R ¹ is -CH ₃ ;

R ³ is -CH ₃ ;

R ^8A and R ^8B are each independently, at each occurrence, -H or -CH ₃ ;

R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci-Ce alkyl,

-C ₃ -Cs cycloalkyl, -Ci-C ₃ alkylene-(C ₃ -Ce cycloalkyl), 4- to 6-membered heterocycloalkyl, - Ci-C ₃ alkylene-(4- to 6-membered heterocycloalkyl), -Ce aryl, -Ci-C ₃ alkylene-Ce aryl, 5- to 6- membered heteroaryl, or -Ci-C ₃ alkylene-(5- to 6-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or two, preferably with exactly one -C2 alkynyl, halogen, -OH, -(OCH2-CH2)I-2-OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or - S(Ci-C ₂ alkyl); wherein said -C ₃ -Ce cycloalkyl is each independently optionally substituted with one or two, preferably with exactly one -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 6-membered heterocycloalkyl is each independently optionally substituted with one or two, preferably with exactly one -C1-C2 alkyl or oxo; wherein said -Ce aryl is each independently optionally substituted with one or two, preferably with exactly one halogen, -OH, or -C1-C2 alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or two -C1-C2 alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl (preferably cyclopropyl), or oxo;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3; m is independently 1 or 2; and n is independently 0 or 1.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci- Ce alkyl, -Cs-Cs cycloalkyl, -Ci-Ce alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, -Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkyl ene-Ce-C 10 aryl, 5- to 10-membered heteroaryl, or -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I-2-OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); wherein said -Cs-Cs cycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), or -Ci-Ce alkoxy; wherein said 5- to 10-membered heteroaryl is independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl, or oxo; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), (I- D3), or (I-D4), R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci- Ce alkyl, -Cs-Cs cycloalkyl, -Ci-Ce alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, -Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkyl ene-Ce-C io aryl, 5- to 10-membered heteroaryl, or -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); wherein said -Cti-Cs cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more halogen, -OH, or -Ci-Ce alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl, or oxo; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci- Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-C8 cycloalkyl), 4- to 8-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -C1-C3 alkyl ene-Ce-C 10 aryl, 5- to 10-membered heteroaryl, or -C1-C3 alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C ₂ alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I. ₂ -OH, -Ci-C ₂ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₂ alkyl); wherein said -C3-C8 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -C1-C2 alkyl or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more halogen, -OH, or -C1-C2 alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -C1-C2 alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl (preferably cyclopropyl), or oxo; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci- Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-C6 cycloalkyl), 4- to 6-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 6-membered heterocycloalkyl), -Ce aryl, -C1-C3 alkylene-Ce aryl, 5- to 6-membered heteroaryl, or -C1-C3 alkylene-(5- to 6-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I-2-OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₂ alkyl); wherein said -C3-C6 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 6-membered heterocycloalkyl is each independently optionally substituted with one or more -C1-C2 alkyl or oxo; wherein said -Ce aryl is each independently optionally substituted with one or more halogen, -OH, or -C1-C2 alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -C1-C2 alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl (preferably cyclopropyl), or oxo;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), (I- D3), or (I-D4), R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci- Ce alkyl, -Cs-Cs cycloalkyl, -C1-C3 alkylene-(C3-Ce cycloalkyl), 4- to 6-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 6-membered heterocycloalkyl), -Ce aryl, -C1-C3 alkylene-Ce aryl, 5- to 6-membered heteroaryl, or -C1-C3 alkylene-(5- to 6-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or two, preferably with exactly one -C2 alkynyl, halogen, -OH, -(OCH2-CH2)i-2-OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or - S(Ci-C ₂ alkyl); wherein said -C3-C6 cycloalkyl is each independently optionally substituted with one or two, preferably with exactly one -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 6-membered heterocycloalkyl is each independently optionally substituted with one or two, preferably with exactly one -C1-C2 alkyl or oxo; wherein said -Ce aryl is each independently optionally substituted with one or two, preferably with exactly one halogen, -OH, or -C1-C2 alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or two -C1-C2 alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl (preferably cyclopropyl), or oxo;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸² is -H or C1-C2 alkyl, preferably R ⁸² is H; R ⁸¹ is independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -Cs-Cs cycloalkyl, -Ci-Ce alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, -Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkylene-Ce-Cio aryl, 5- to 10-membered heteroaryl, or -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )i-2-OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); wherein said -Cs-Cs cycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), or -Ci-Ce alkoxy; wherein said 5- to 10-membered heteroaryl is independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl, or oxo; and R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸² is -H or C1-C2 alkyl, preferably R ⁸² is H; R ⁸¹ is independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -Cs-Cs cycloalkyl, -Ci-Ce alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, -Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkylene-Ce-Cio aryl, 5- to 10-membered heteroaryl, or -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I-2-OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); wherein said -Cs-Cs cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more halogen, -OH, or -Ci-Ce alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl, or oxo; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), (I- D3), or (I-D4), R ⁸² is -H or C1-C2 alkyl, preferably R ⁸² is H; R ⁸¹ is independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -C1-C3 alkylene-Ce-Cio aryl, 5- to 10-membered heteroaryl, or -C1-C3 alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₂ alkyl); wherein said -C3-C8 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -C1-C2 alkyl or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more halogen, -OH, or -C1-C2 alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -C1-C2 alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl (preferably cyclopropyl), or oxo; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸² is -H or C1-C2 alkyl, preferably R ⁸² is H; R ⁸¹ is independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-C6 cycloalkyl), 4- to 6-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 6-membered heterocycloalkyl), -Ce aryl, -C1-C3 alkylene-Ce aryl, 5- to 6-membered heteroaryl, or -C1-C3 alkyl ene-(5- to 6-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )i-2-OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₂ alkyl); wherein said -C3-C6 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 6-membered heterocycloalkyl is each independently optionally substituted with one or more -C1-C2 alkyl or oxo; wherein said -Ce aryl is each independently optionally substituted with one or more halogen, -OH, or -C1-C2 alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -C1-C2 alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl (preferably cyclopropyl), or oxo;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), (I- D3), or (I-D4), R ⁸² is -H or C1-C2 alkyl, preferably R ⁸² is H; R ⁸¹ is independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -Cs-Cs cycloalkyl, -C1-C3 alkylene-(C3-Ce cycloalkyl), 4- to 6-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 6-membered heterocycloalkyl), -Ce aryl, -C1-C3 alkylene-Ce aryl, 5- to 6-membered heteroaryl, or -C1-C3 alkyl ene-(5- to 6-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or two, preferably with exactly one -C2 alkynyl, halogen, -OH, -(OCH2-CH2)I-2-OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or - S(Ci-C ₂ alkyl); wherein said -C3-C6 cycloalkyl is each independently optionally substituted with one or two, preferably with exactly one -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 6-membered heterocycloalkyl is each independently optionally substituted with one or two, preferably with exactly one -C1-C2 alkyl or oxo; wherein said -Ce aryl is each independently optionally substituted with one or two, preferably with exactly one halogen, -OH, or -C1-C2 alkoxy; or R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or two -C1-C2 alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl (preferably cyclopropyl), or oxo;

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci- Ce alkyl, -Cs-Cs cycloalkyl, -Ci-Ce alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, -Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkyl ene-Ce-C 10 aryl, 5- to 10-membered heteroaryl, or -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )i-2-OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); wherein said -Cti-Cs cycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), or -Ci-Ce alkoxy; wherein said 5- to 10-membered heteroaryl is independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci- Ce alkyl, -Cs-Cs cycloalkyl, -Ci-Ce alkylene-(C3-C8 cycloalkyl), 4- to 8-membered heterocycloalkyl, -Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkyl ene-Ce-C 10 aryl, 5- to 10-membered heteroaryl, or -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I-2-OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); wherein said -C3-C8 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more halogen, -OH, or -Ci-Ce alkoxy; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), (I- D3), or (I-D4), R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci- Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-C8 cycloalkyl), 4- to 8-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -C1-C3 alkyl ene-Ce-C 10 aryl, 5- to 10-membered heteroaryl, or -C1-C3 alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C ₂ alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -Ci-C ₂ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₂ alkyl); wherein said -C3-C8 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -C1-C2 alkyl or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more halogen, -OH, or -C1-C2 alkoxy; and R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci- Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-C6 cycloalkyl), 4- to 6-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 6-membered heterocycloalkyl), -Ce aryl, -C1-C3 alkylene-Ce aryl, 5- to 6-membered heteroaryl, or -C1-C3 alkylene-(5- to 6-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I-2-OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₂ alkyl); wherein said -C3-C6 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 6-membered heterocycloalkyl is each independently optionally substituted with one or more -C1-C2 alkyl or oxo; wherein said -Ce aryl is each independently optionally substituted with one or more halogen, -OH, or -C1-C2 alkoxy; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), (I- D3), or (I-D4), R ⁸¹ and R ⁸² are each independently, at each occurrence, selected from -H, -Ci- Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-C6 cycloalkyl), 4- to 6-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 6-membered heterocycloalkyl), -Ce aryl, -C1-C3 alkylene-Ce aryl, 5- to 6-membered heteroaryl, or -C1-C3 alkylene-(5- to 6-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or two, preferably with exactly one -C2 alkynyl, halogen, -OH, -(OCH2-CH2)i-2-OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or - S(Ci-C ₂ alkyl); wherein said -C3-C6 cycloalkyl is each independently optionally substituted with one or two, preferably with exactly one -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 6-membered heterocycloalkyl is each independently optionally substituted with one or two, preferably with exactly one -C1-C2 alkyl or oxo; wherein said -Ce aryl is each independently optionally substituted with one or two, preferably with exactly one halogen, -OH, or -C1-C2 alkoxy; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸² is -H or C1-C2 alkyl, preferably R ⁸² is H; R ⁸¹ is independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C3-C8 cycloalkyl, -Ci-Ce alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, -Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkylene-Ce-Cio aryl, 5- to 10-membered heteroaryl, or -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I-2-OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); wherein said -C3-C8 cycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), or -Ci-Ce alkoxy; wherein said 5- to 10-membered heteroaryl is independently optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -N(R ⁸³ )(R ⁸⁴ ), -Ci-Ce alkoxy, phenyl, or oxo; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), (I- D3), or (I-D4), R ⁸² is -H or C1-C2 alkyl, preferably R ⁸² is H; R ⁸¹ is independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C3-C8 cycloalkyl, -Ci-Ce alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, -Ci-Ce alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -Ci-Ce alkylene-Ce-Cio aryl, 5- to 10-membered heteroaryl, or -Ci-Ce alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2-C6 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -Ci-C ₆ alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₆ alkyl); wherein said -C3-C8 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -Ci-Ce alkyl or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more halogen, -OH, or -Ci-Ce alkoxy; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸² is -H or C1-C2 alkyl, preferably R ⁸² is H; R ⁸¹ is independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-Cs cycloalkyl), 4- to 8-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 8-membered heterocycloalkyl), -Ce-Cio aryl, -C1-C3 alkylene-Ce-Cio aryl, 5- to 10-membered heteroaryl, or -C1-C3 alkylene-(5- to 10-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I-2-OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₂ alkyl); wherein said -C3-C8 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 8-membered heterocycloalkyl is each independently optionally substituted with one or more -C1-C2 alkyl or oxo; wherein said -Ce-Cio aryl is each independently optionally substituted with one or more halogen, -OH, or -C1-C2 alkoxy; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), (I- D3), or (I-D4), R ⁸² is -H or C1-C2 alkyl, preferably R ⁸² is H; R ⁸¹ is independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-C6 cycloalkyl), 4- to 6-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 6-membered heterocycloalkyl), -Ce aryl, -C1-C3 alkylene-Ce aryl, 5- to 6-membered heteroaryl, or -C1-C3 alkyl ene-(5- to 6-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or more -C2 alkynyl, halogen, -OH, -(OCH ₂ -CH ₂ )I- ₂ -OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or -S(Ci-C ₂ alkyl); wherein said -C3-C6 cycloalkyl is each independently optionally substituted with one or more -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 6-membered heterocycloalkyl is each independently optionally substituted with one or more -C1-C2 alkyl or oxo; wherein said -Ce aryl is each independently optionally substituted with one or more halogen, -OH, or -C1-C2 alkoxy; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), CIDS), or (I-D4), R ⁸² is -H or C1-C2 alkyl, preferably R ⁸² is H; R ⁸¹ is independently, at each occurrence, selected from -H, -Ci-Ce alkyl, -C3-C8 cycloalkyl, -C1-C3 alkylene-(C3-C6 cycloalkyl), 4- to 6-membered heterocycloalkyl, -C1-C3 alkylene-(4- to 6-membered heterocycloalkyl), -Ce aryl, -C1-C3 alkylene-Ce aryl, 5- to 6-membered heteroaryl, or -C1-C3 alkyl ene-(5- to 6-membered heteroaryl); wherein said -Ci-Ce alkyl is optionally substituted with one or two, preferably with exactly one -C2 alkynyl, halogen, -OH, -(OCH2-CH2)I-2-OH, -C1-C2 alkoxy, -N(R ⁸³ )(R ⁸⁴ ), or - S(Ci-C ₂ alkyl); wherein said -C3-C6 cycloalkyl is each independently optionally substituted with one or two, preferably with exactly one -OH, -N(R ⁸³ )(R ⁸⁴ ), or phenyl; wherein said 4- to 6-membered heterocycloalkyl is each independently optionally substituted with one or two, preferably with exactly one -C1-C2 alkyl or oxo; wherein said -Ce aryl is each independently optionally substituted with one or two, preferably with exactly one halogen, -OH, or -C1-C2 alkoxy; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), (I- D3), or (I-D4), R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -Ci-Ce alkyl, halogen, -OH, -C1-C3 alkylene-OH, -N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl, or oxo; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and -Ci-Ce alkyl.

In some embodiments of any of Formulae (I), (I-A), (I-Al), (I-A2), (I- A3), (I-A4), (I- B), (I-Bl), (I-B2), (I-B3), (I-B4), (I-C), (I-Cl), (I-C2), (I-C3), (I-C4), (I-D), (I-Dl), (I-D2), (I- D3), or (I-D4),

R ⁸¹ and R ⁸² , together with the nitrogen atom to which they are attached, combine to form a 4- to 11 -membered heterocycloalkyl, wherein said 4- to 11 -membered heterocycloalkyl is optionally substituted with one or more -C1-C2 alkyl, halogen, -OH, -C1-C3 alkylene-OH, - N(R ⁸³ )(R ⁸⁴ ), -C1-C3 alkylene-N(R ⁸³ )(R ⁸⁴ ), -C3-C6 cycloalkyl (preferably cyclopropyl), or oxo; and

R ⁸³ and R ⁸⁴ are independently, at each occurrence, selected from -H and C1-C2 alkyl, preferably -H and -CH3.

In preferred embodiments, said compound is selected from:

In some embodiments, said compound is 3-(2,4-dimethylphenyl)sulfonyl-8-[4-(2-oxo- 2-pyrrolidin- 1 -yl-ethy l)piperazin- 1 -yl] -4H-triazolo [ 1 ,5 -a] quinazolin-5 -one (Compound 1 ).

In some embodiments, said compound is 3-(2,4-dimethylphenyl)sulfonyl-8-[4-(2- morpholino-2-oxo-ethyl)piperazin-l -yl]-4H-triazolo[l,5-a]quinazolin-5-one (Compound 2).

Anti- virulence Activity of the Inventive Compounds

The inventive compounds are 3-(phenylsulfonyl)-[l,2,3]triazolo[l,5a]quinazolin- 5(4h)-one derivatives comprising an N-bound piperazine or homopiperazine at the 8-position of the [l,2,3]triazolo[l,5a]quinazolin-5(4h)-one core, further substituted (on the second nitrogen atom) by an (optionally substituted) acetyl or propionyl amide group. As shown in Table 3, the inventive compounds were tested in an AgrA reporter assay to evaluate their ability to inhibit the expression of genes under control of the P3 promoter, which is regulated by AgrA. Specifically, the reporter gene lacZ (which encodes the protein product B-galactosidase) was transfected into 5. aureus with a plasmid under control of the P3 promoter. The compounds were serially diluted in lysogeny broth (LB), and incubated with 5. aureus. Following incubation, the 5. aureus cells were lysed and treated with 4-methylumbelliferyl- galactopyranoside (MUG). The B-galactosidase produced by expression of lacZ cleaved the MUG into galactopyranose and 4-methylumbelliferone, which was measured by fluorescence. The fluorescence readout thus serves as a proxy for the ability of the inventive compounds to inhibit genes under the control of the P3 promoter (e.g., in bacteria, preferably in S. aureus).

The inventive compounds retained activity in the presence of 40 mg/mL HSA with IC50 values as low as 8 pg/mL. Thus, the inventive compounds retained activity to inhibit expression of AgrA-regulated genes even when incubated with physiologically relevant concentrations of - 1Q -

HSA. In contrast, and as shown in Example 3, even though Compound 45 of WO 2020/109350 inhibited the expression of lacZ at a concentration of less than 1 pg/mL in the AgrA reporter assay in the absence of HSA, when incubated with 5. aureus in the presence of HSA, the compound was inactive even at the highest concentration of compound tested, i.e., 128 pg/mL. Without wishing to be bound, the concentration of 40 mg/mL HSA was chosen because this is the same concentration of HSA found in human serum.

Without wishing to be bound, the ability of the inventive compounds to retain their activity even in the presence of HSA enables the inventive compounds to be administered (e.g., to a subject such as a mammal, preferably a human; and preferably topically) while still maintaining efficacy. Thus, the inventive compounds were surprisingly able to maintain their activity even when incubated in biologically relevant concentrations of HSA.

Use of the Inventive Compounds

In one aspect, the present invention provides a pharmaceutical composition comprising at least one compound according to Formula (I), or a pharmaceutically acceptable salt, tautomer, solvate or hydrate thereof, and a pharmaceutically acceptable excipient.

In one aspect, the present invention provides a compound according to Formula (I) or a pharmaceutically acceptable salt, tautomer, solvate or hydrate thereof, or a pharmaceutical composition comprising a compound of Formula (I), for use as a medicament.

In a further aspect, the present invention provides a compound according to Formula (I) and pharmaceutically acceptable salts, stereoisomers, enantiomers, tautomers of the compounds of Formula (I) as disclosed herein for use in a method of reducing the virulence of bacteria, preferably of bacteria expressing AgrA or an ortholog of AgrA, preferably AgrA, and further preferably of bacteria of the genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably of Staphylococcus aureus. In a further very preferred embodiment, said bacteria expresses AgrA or an ortholog of AgrA. In a further very preferred embodiment, said bacteria expresses AgrA. In a further very preferred embodiment, said bacteria is of the genus selected from Staphylococcus, Streptococcus or Clostridium. In a further very preferred embodiment, said bacteria is of the genus Staphylococcus. In a further very preferred embodiment, said bacteria is Staphylococcus aureus. The herein described and disclosed embodiments, preferred embodiments and very preferred embodiments for the compounds of Formula (I) should apply to this inventive method irrespective of whether is specifically referred to again or its repetition is avoided for the sake of conciseness. In a further aspect, the present invention provides a compound according to Formula (I) and pharmaceutically acceptable salts, stereoisomers, enantiomers, tautomers of the compounds of Formula (I) as disclosed herein for use in a method of preventing or treating a disease in a subject, preferably an infection or an inflammatory disease, further preferably a bacterial infection or an inflammatory skin disease, caused or exacerbated by bacteria, wherein preferably said bacteria is selected from the genus of Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably wherein said bacteria is Staphylococcus aureus. In a further very preferred embodiment, said disease is an infection or an inflammatory disease. In a further very preferred embodiment, said disease is an infection. In a further very preferred embodiment, said disease is a bacterial infection or an inflammatory skin disease, caused or exacerbated by bacteria. In a further very preferred embodiment, said disease a bacterial infection, wherein said bacteria is selected from the genus of Staphylococcus, Streptococcus or Clostridium. In a further very preferred embodiment, said disease is exacerbated by bacteria of the genus selected from Staphylococcus, preferably from Staphylococcus aureus. In a further very preferred embodiment, said disease is an infection caused or exacerbated by bacteria selected from the genus of Staphylococcus. In a further very preferred embodiment, said disease is an infection caused or exacerbated by Staphylococcus aureus. In a further very preferred embodiment, said disease is Netherton syndrome or an inflammatory skin disease, preferably atopic dermatitis. In a further very preferred embodiment, said disease is an inflammatory skin disease, preferably atopic dermatitis exacerbated by Staphylococcus aureus. In a preferred embodiment, said disease is Netherton syndrome. The herein described and disclosed embodiments, preferred embodiments and very preferred embodiments for the compounds of formula I should apply to this inventive method irrespective of whether is specifically referred to again or its repetition is avoided for the sake of conciseness.

In a further aspect, the present invention provides a compound according to Formula (I) for use in a method of inhibition of the quorum sensing, preferably AgrA quorum sensing, in bacteria, preferably in bacteria of the genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably in Staphylococcus, and again further preferably in Staphylococcus aureus. The herein described and disclosed embodiments, preferred embodiments and very preferred embodiments for the compounds of formula I should apply to this inventive method irrespective of whether is specifically referred to again or its repetition is avoided for the sake of conciseness.

In a further preferred embodiment, said bacteria is selected from Streptococcus pyogenes, Clostridium difficile or Staphylococcus aureus, preferably said bacteria is Staphylococcus aureus. In a very preferred embodiment, said bacteria is Staphylococcus aureus.

Thus, the inventive compositions and compounds represent anti-virulence therapeutics which may be used as standalone therapy to strengthen the infected host’s self-defense and - healing capacity through reduction of tissue damage, reduction in inflammation, reduction in disease dissemination, full immune response and reduction in reoccurrence rate. In cases where the disarmed pathogen may not be sufficiently cleared by the host, a combination therapy with traditional antibiotics (for infections caused by 5. aureus) or with anti-inflammatory agents (for diseases exacerbated by 5. aureus) might be considered, allowing eventually to optimize the timeliness and/or the dose of therapy (Dickey SW, et al. (2017) Nat Rev Drug Discov 16(7):457-471).

In a preferred embodiment of the inventive method, said compound of Formula (I) inhibits the synthesis of one or more virulence factors by the bacteria, wherein said one or more virulence factor is selected from the group consisting of one or more of toxins (e.g., a, p, y, y- variant, and 8-hemolysins, PSMs (e.g., PSMa), Panton-Valentine leukocidin (PVL), leukotoxin E and D (LukED), leukotoxin G and H (LukGH), enterotoxins (e.g., enterotoxin B), exfoliative toxin), proteases (e.g., serine proteases, metalloproteases and cysteine proteases), nuclease, lipase, coagulase, hyaluronidase, clumping factor, pyrogenic toxin superantigen (e.g., TSST- 1), and combinations thereof. Thus, in a further preferred embodiment of the inventive method, said one toxin is selected from a, , y, y-variant, and 8-hemolysins. In a further preferred embodiment of the inventive method, said PSM is PSMa. In a further preferred embodiment of the inventive method, said enterotoxin is enterotoxin B or exfoliative toxin. In a further preferred embodiment of the inventive method, said protease is selected from serine proteases, metalloproteases and cysteine proteases. In a further preferred embodiment of the inventive method, said pyrogenic toxin superantigen is TSST-1. In a further preferred embodiment of the inventive method, said compound of Formula (I) inhibits the synthesis of one or more virulence factors by the bacteria, wherein said one or more virulence factor is selected from the group consisting of a, p, y, y-variant, and 8-hemolysin, PSMa, Panton-Valentine leukocidin (PVL), leukotoxin E and D (LukED), leukotoxin G and H (LukGH), enterotoxin B, exfoliative toxin, a serine protease, a metalloprotease, a cysteine protease, a nuclease, a lipase, a coagulase, a hyaluronidase, a clumping factor, TSST-1, and any combination of one or more of any specific virulence factor or generic group of virulence factors thereof. In a further very preferred embodiment of the inventive method, said compound of Formula (I) inhibits the expression of PSMa, RNAIII and/or any of its downstream targets. In a further very preferred embodiment of the inventive method, said compound of Formula (I) inhibits the expression of PSMa. In a further very preferred embodiment of the inventive method, said compound of Formula (I) inhibits the expression of RNAIII. In a further very preferred embodiment of the inventive method, said compound of Formula (I) inhibits the expression of a downstream target of RNAIII. In a further very preferred embodiment, said method further comprises administering an antibiotic or an anti-inflammatory agent to said subject, preferably to said human. In a further very preferred embodiment, said method further comprises administering an antibiotic to said subject, preferably to said human. In a further very preferred embodiment, said method further comprises administering an anti-inflammatory agent, preferably to said human.

In a further aspect, the present invention provides for a method of preventing or treating a disease of a subject, preferably an infection or an inflammatory disease, further preferably a bacterial infection or an inflammatory skin disease, caused or exacerbated by bacteria, wherein preferably said bacteria is selected from the genus of Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably wherein said bacteria is Staphylococcus aureus, said method comprises administering to the subject in need of such prevention or treatment an effective amount of a compound of Formula (I), a pharmaceutical composition or a combination product, preferably a compound of Formula (I). In a further very preferred embodiment, said disease is an infection or an inflammatory disease. In a further very preferred embodiment, said disease is an infection. In a further very preferred embodiment, said disease is a bacterial infection. In a further very preferred embodiment, said disease is an inflammatory skin disease caused or exacerbated by bacteria. Preferably said infection, preferably said bacterial infection is caused by bacteria of the genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably by Staphylococcus, and again further preferably by Staphylococcus aureus. In a further very preferred embodiment, saidaid infection, preferably said bacterial infection comprises an antibiotic resistant Staphylococcus infection, preferably said antibiotic resistant Staphylococcus infection comprises a Methicillin- resistant Staphylococcus aureus infection.

Thus, an inventive compound of Formula (I) or an inventive composition, preferably an inventive pharmaceutical composition, comprising an inventive compound of Formula (I), as described herein, can be administered to a subject to inhibit the activity of AgrA thereby preventing the production of virulence factors that aid in bacterial infection or development of a disease condition or disorder associated with the bacterial infection. Examples of diseases and disorders associated with a bacterial infection responsive to treatment with the inventive compounds and/or compositions can include, without limitation, skin and soft tissue infections, lung infections or chronic inflammatory skin diseases such as atopic dermatitis or Netherton syndrome.

In a further aspect, the present invention provides a method for inhibiting the quorum sensing, preferably AgrA quorum sensing, in bacteria, preferably in bacteria of the genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably in Staphylococcus, and again further preferably in Staphylococcus aureus, wherein said comprises administering to a subject an effective amount of a compound of Formula (I), a pharmaceutical composition or a combination product, preferably a compound of Formula (I), in accordance.

In a further preferred embodiment of the present invention, said compound of Formula (I) is provided in a topical composition with a pharmaceutically acceptable carrier and is topically administered to a subject, wherein preferably subject has a disease or disorder associated with a bacterial infection, wherein said bacterial infection is an infection caused or exacerbated by bacteria, preferably by Staphylococcus aureus. In a further preferred embodiment said infection caused or exacerbated by bacteria is SSTI, Netherton syndrome or atopic dermatitis. In a further preferred embodiment said bacterial infection is SSTI, Netherton syndrome or atopic dermatitis.

Thus, an inventive compound of Formula (I) or an inventive composition or an inventive combination product, preferably an inventive compound of Formula (I) inventive pharmaceutical composition, comprising an inventive compound of Formula (I), as described herein, can be used to prevent or treat infection of a subject by any bacteria species that utilizes the AgrA response regulator in quorum sensing and the production of virulence factors. The inventive compounds and compositions are typically and preferably administered to subjects having or at risk of having an infection, preferably a bacterial infection such as a. Staphylococcus and/or Streptococcus infection. For example, a subject that can benefit from treatment with an inventive compound or composition, as described herein, can be a hospital patient at risk of developing nosocomial infection or a subject known to be infected with or having been exposed to antibiotic resistant bacteria such as, for example, Methicillin-resistant 5. aureus, Vancomycin-intermediary-sensible S. aureus, and Vancomycin-resistant S. aureus. Methods of detecting the presence of a Staphylococcus bacterial infection are well known, for example, by culturing from a sample from the subject, e.g. a blood culture, can be used.

In a further aspect, the present invention provides a method for the treatment of a subject suffering from an infection or an inflammatory disease, preferably a bacterial infection or an inflammatory skin disease, caused by bacteria of the genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably by Staphylococcus, and again further preferably by Staphylococcus aureus, said method comprising administering to the subject a compound of Formula (I) in accordance with the present invention and at least one antibiotic active against bacteria, preferably against bacteria of the genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably against Staphylococcus, and again further preferably against Staphylococcus aureus.

In a further aspect, the present invention provides a method for preventing or treating a bacterial infection in a subject comprising the step of administering an effective amount of a compound of Formula (I) in accordance with the present invention to said subject in need of such prevention or treatment. Preferably said bacterial infection is caused by bacteria of the genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably by Staphylococcus, and again further preferably by Staphylococcus aureus.

In a further aspect, the present invention provides a method of preventing or treating a disease caused or exacerbated by Gram-positive quorum sensing bacteria, comprising administering to a subject an effective amount of a compound of Formula (I) in accordance with the present invention to a subject in need of such prevention or treatment.

In one aspect, the present disclosure provides a pharmaceutical composition comprising at least one compound according to Formula (I), or a pharmaceutically acceptable salt, tautomer, solvate or hydrate thereof, and a pharmaceutically acceptable excipient.

In one aspect, the present disclosure provides a compound according to Formula (I) or a pharmaceutically acceptable salt, tautomer, solvate or hydrate thereof, or a pharmaceutical composition comprising a compound according to Formula (I), for use as a medicament.

In one aspect, the invention provides an inventive compound as described herein or a pharmaceutical composition comprising an inventive compound as described herein for use in a method of reducing the virulence of a bacteria, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA.

In one aspect, the invention provides an inventive compound as described herein or a pharmaceutical composition comprising an inventive compound as described herein for use in a method of inhibiting the quorum sensing in bacteria, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA.

In one aspect, the invention provides an inventive compound as described herein or a pharmaceutical composition comprising an inventive compound as described herein for use in a method of preventing or treating diseases caused or exacerbated by bacteria in a subject, preferably a bacteria of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the disease is a skin disease or a lung disease, more preferably wherein the skin disease is atopic dermatitis, Netherton syndrome or psoriasis.

In some embodiments, the invention provides the use of an inventive compound as described herein or the use of a pharmaceutical composition comprising an inventive compound as described herein in a method of reducing the virulence of a bacteria, preferably in a subject in need thereof, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA.

In some embodiments, the invention provides the use of an inventive compound as described herein or the use of a pharmaceutical composition comprising an inventive compound as described herein in a method of inhibiting the quorum sensing in bacteria, preferably in a subject in need thereof, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA.

In some embodiments, the invention provides the use of an inventive compound as described herein or the use of a pharmaceutical composition comprising an inventive compound as described herein in a method of preventing or treating diseases caused or exacerbated by bacteria, preferably in a subject in need thereof, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the disease is a skin disease or a lung disease, more preferably wherein the skin disease is atopic dermatitis, Netherton syndrome or psoriasis.

In some embodiments, the invention provides the use of an inventive compound as described herein or the use of a pharmaceutical composition comprising an inventive compound as described herein in the manufacture of a medicament for reducing the virulence of a bacteria, preferably in a subject in need thereof, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA.

In some embodiments, the invention provides the use of an inventive compound as described herein or the use of a pharmaceutical composition comprising an inventive compound as described herein in the manufacture of a medicament for inhibiting the quorum sensing in bacteria, preferably in a subject in need thereof, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA.

In some embodiments, the invention provides the use of an inventive compound as described herein or the use of a pharmaceutical composition comprising an inventive compound as described herein in the manufacture of a medicament for preventing or treating diseases caused or exacerbated by bacteria, preferably in a subject in need thereof, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the disease is a skin disease or a lung disease, more preferably wherein the skin disease is atopic dermatitis, Netherton syndrome or psoriasis.

In some embodiments, the invention provides a method of treating a bacterial infection in a subject in need thereof, the method comprising administering to the subject an inventive compound as described herein or a pharmaceutical composition comprising an inventive composition as described herein, preferably wherein the virulence of the bacteria is reduced; preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA.

In some embodiments, the invention provides a method of treating a bacterial infection in a subject in need thereof, the method comprising administering to the subject an inventive compound as described herein or a pharmaceutical composition comprising an inventive composition as described herein, preferably wherein the quorum sensing of the bacteria is inhibited, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA.

In some embodiments, the invention provides a method of treating a disease caused or exacerbated by a bacterial infection in a subject in need thereof, the method comprising administering to the subject an inventive compound as described herein or a pharmaceutical composition comprising an inventive composition as described herein, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the disease is a skin disease or a lung disease, more preferably wherein the skin disease is atopic dermatitis, Netherton syndrome or psoriasis.

In some embodiments, the invention provides a method of reducing the virulence of a bacteria, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA, comprising exposing the bacteria to an inventive compound as described herein or a pharmaceutical composition comprising an inventive compound as described herein.

In some embodiments, the invention provides a method of inhibiting the quorum sensing in bacteria, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the bacteria expresses AgrA or an ortholog of AgrA, more preferably wherein the bacteria expresses AgrA, comprising exposing the bacteria to an inventive compound as described herein or a pharmaceutical composition comprising an inventive compound as described herein.

In some embodiments, the invention provides a method of preventing or treating diseases caused or exacerbated by bacteria, preferably wherein the bacteria is of a genus selected from Staphylococcus, Streptococcus or Clostridium, more preferably of Staphylococcus, and again further preferably is Staphylococcus aureus, preferably wherein the disease is a skin disease or a lung disease, more preferably wherein the skin disease is atopic dermatitis, Netherton syndrome or psoriasis, comprising exposing the bacteria to an inventive compound as described herein or a pharmaceutical composition comprising an inventive compound as described herein.

The inventive compounds of Formula (I), pharmaceutical compositions comprising the same or combination products can be administered to any subject that can experience the beneficial effects of the inventive compounds, compositions or products, as described herein. Preferably said subject is a human. The inventive compounds, compositions comprising the same or products as described herein can be administered by any means that achieve their intended purpose. For example, administration can be local or systemic, e.g., administration can be parenteral, topical, local, subcutaneous, oral, intravenous, intraarticular, intrathecal, intramuscular, intraperitoneal, intradermal, transdermal, buccal, oromucosal, or ocular, or administration via inhalation. Equivalents

While the present technology has been described in conjunction with the specific embodiments set forth above, many alternatives, modifications and other variations thereof will be apparent to those of ordinary skill in the art. All such alternatives, modifications and variations are intended to fall within the spirit and scope of the present invention.

EXAMPLES

The invention will now be illustrated by way of the following non-limiting examples.

While particular embodiments of the invention are described below a skilled person will appreciate that various changes and modifications can be made. References to preparations carried out in a similar manner to, or by the general method of, other preparations, may encompass variations in routine parameters such as time, temperature, workup conditions, minor changes in reagents amounts, and the like. Abbreviations

The following list provides definitions of certain abbreviations and symbols as used herein. It will be appreciated that the list is not exhaustive, but the meaning of those abbreviations and symbols not herein below defined will be readily apparent to those skilled in the art. In describing the invention, chemical elements are identified in accordance with the Periodic Table of the Elements.

HPLC-MS

Unless specified otherwise, the purity and identity of Intermediate or Example compounds were assessed by state-of-the-art HLPC-MS as described below. HPLC-MS analysis was performed on LC-MS Waters Alliance Micromass ZQ 2000 system equipped with a Waters 2747 sample manager, a Waters 2695 separations module, a Waters 2996 photodiode array detector (200-800 nm) and a Waters Micromass ZQ2000 detector (scant 00-800). XBridge Cl 8 column (3.5 pm particle size, dimensions 50 x 4.6 mm) was used for HPLC analysis. The injection volume was 20 pL. A mixture of water and acetonitrile was used as mobile phase in gradient-elution. The pH of the mobile phase was adjusted with HCOOH and NH4OH to form a buffer solution at pH 3.8. The analysis time was 5 min (Method Al) or 10 min (Method A2) using a gradient starting from 100% aqueous buffer pH = 3.8 ± 0.3 (NH4OH 0.01%, HCOOH 0.02%) and reaching 100% acetonitrile buffer (NH ₄ OH 0.01%, HCOOH 0.02%) - within 3.25 min at a flow rate of 2 mL/min (5 min run; Method Al), within 6.5 min at a flow rate of 2 mL/min (10 min run; Method A2).

Preparative Example 1: 8-bromo-3-(2.4-dimethylphenyl)sulfonyl-4H-triazolo[l,5-

To an ice-cooled suspension of 2,4-dimethylbenzenethiol (9.8ml, 72.46mmol) and potassium carbonate (20g, 144.7mmol) in dry DMF (200ml) was added 2-bromoacetonitrile (76.09 mmol). The reaction mixture was stirred for 1.5h (on ice). Water (350ml) was then added, and the product was extracted with Et20 (3x100ml). The organic layers were combined, washed with brine (3x80ml), dried over MgSO4, filtered and evaporated to afford the intermediate 2-(2,4-dimethylphenyl)-sulfanylacetonitrile as a colourless oil (99.8% yield, >99% purity by HPLC).

The previous intermediate was solubilized in DCM (250ml) and cooled down on an ice bath. mCPBA (<77%, 33g, 147.2mmol) was then added portionwise. The reaction mixture was stirred for 40min on ice and was left at rt overnight. The reaction mixture was then diluted with DCM (250ml), washed with 5% aq. Na2S20s (2x100ml) and cone. aq. NaHCOs (8x200ml). The combined aqueous were extracted with DCM (2x100ml). Organic phases were combined and dried over MgSO4, filtered and evaporated to yield 2-(2,4-dimethylphenyl)-sulfonylacetonitrile as a white powder (92.1% yield, 98% purity by HPLC).

To a cooled (4°C) solution of methyl 2-amino-4-bromo-benzoate (7g, 30.4 mmol) in dry acetonitrile, under Ar atmosphere, tert-butyl nitrite (1.5 eq.) was added dropwise. After 20 min stirring, azidotrimethylsilane (1.2 eq.) was added dropwise (exothermic reaction - reaction kept below 15°C). After addition was completed, the reaction mixture was stirred for 40min in an ice bath and then allowed to reach rt and stirred for another 2h. Concentrated aq. NaHCCL solution (500ml) was added to the reaction mixture leading to a suspension which was extracted with EtOAc (3x200ml). The combined organic layers were washed with brine (200ml) dried over MgSCL, filtered and evaporated to afford methyl 2-azido-4-bromo-benzoate as an orange oil (97% yield, 99% purity by HPLC).

To an ice-cooled solution of 2-(2,4-dimethylphenyl)-sulfonylacetonitrile (11.67g, 54.65mmol) in dry EtOH (560ml) was added dropwise a freshly prepared solution of sodium ethoxide in EtOH (15%, 117mmol). After 30min, a solution of methyl 2-azido-4-bromo- benzoate (54.13mmol) in dry EtOH (225ml) was added dropwise. The resulting mixture was stirred on ice for Ih and then 1.5h at rt and then concentrated under vacuum. pH was adjusted to 3.1 with aq. hydrochloric acid (2M). The precipitate was filtered off. The solid was washed with water (3x400ml) and dried under vacuum. The crude product was triturated several times with cold EtOAc and Et20 and dried overnight to yield the title Intermediate 1-1 as a beige solid (86.6% yield, 99.1% purity by HPLC).

Preparative Example 2: 2-[4-[3-(2.4-dimethylphenyl)sulfonyl-5-oxo-4H-triazolo[l,5- a]quinazolin-8-yl]piperazin-l-yl]acetic acid (Intermediate II-l)

Intermediate II- 1 was prepared in at least two ways from Intermediate 1-1 following the general scheme below:

Route B

>

2. Saponification

Route A:

A 100 ml round -bottom flask was charged with Intermediate 1-1 (600 mg, 1.38 mmol), tert-butyl piperazine- 1 -carboxylate (516 mg, 2.77 mmol, 2 eq), Pd2dbas (39.8 mg, 5 mol %), CS2CO3 (677 mg, 2.08 mmol, 1.5 eq), RuPhos (64.6 mg, 0.138 mmol, 10 mol %) and DMF (40 ml) was added. The mixture was degassed by Ar flush then equipped with a reflux condenser and stirred in a preheated heating block at 120°C for 3h. After this time the mixture was cool down to rt, the mixture was diluted with MeOH, filtered on celite and evaporated to dryness. The crude product was purified by flash chromatography with EtOAc-MeOH (100-0 to 95-5) to afford 750 mg of brownish solid, which were triturated in Et20 to yield 514 mg of the tert-butyl 4-[3-(2,4-dimethylphenyl)sulfonyl-5-oxo-4H-triazolo[l,5-a]qu inazolin-8- yl]piperazine-l -carboxylate as a beige solid (68% yield, 99% purity by HPLC, Method Al). MS (ES+) m/z 539.

To a solution of tert-butyl 4-[3-(2,4-dimethylphenyl)sulfonyl-5-oxo-4H-triazolo[l,5- a]quinazolin-8-yl]piperazine-l-carboxylate (500 mg, 0.928 mmol) in 20 ml of dioxane was added HC1 4M in dioxane (3.45 ml, 15 eq) and the mixture was stirred at 80°C overnight. The precipitate was filtered and triturated with Et20 to afford 410 mg of 3 -(2,4- dimethylphenyl)sulfonyl-8-piperazin-l-yl-4H-triazolo[l,5-a]q uinazolin-5-one as a beige solid (93% yield, 100% purity by HPLC, Method Al). MS (ES+) m/z 439.

Intermediate II- 1 was then prepared in two additional steps, starting from the previous compound (3-(2,4-dimethylphenyl)sulfonyl-8-piperazin-l-yl-4H-triazolo [l,5-a]quinazolin-5- one and methyl bromoacetate in the presence of sodium bicarbonate in acetone, followed by saponification of the ester in the presence of lithium hydroxide as described in route B, below.

Alternatively, Intermediate II- 1 was prepared in two additional steps starting from (3- (2,4-dimethylphenyl)sulfonyl-8-piperazin-l -yl-4H-triazolo[l,5-a]quinazolin-5-one and methyl bromoacetate (3.0 eq) in the presence of potassium carbonate in DMSO. This mixture was stirred at 120°C overnight, then cooled to room temperature. DMSO was removed by freeze drying and the crude product was purified by flash chromatography with ethyl acetate/MeOH (100 to 80-20) to afford a beige solid. Subsequent saponification of the ester in the presence of lithium hydroxide was performed as described in route B, below.

Route B:

In a sealed vial, methyl 2-piperazin-l-ylacetate dihydrochloride (80 mg, 0.35 mmol,l eq), CS2CO3 (395 mg, 1.21mmol, 3.5 eq) in dry DMF (9 ml) was stirred for 2 h. Then Tris(dihenzylideneacetone)dipalladium(0) (9.95 mg, 5mol%), 2-Dicyclohexylphosphino-2',6'- di-i-propoxy-l,l'-biphenyl (16.2 mg, 10 mol%), 1-1 (150 mg, 0.35 mmol) was added to the vial flushed with Ar . The vial was then sealed and stirred in a preheated heating block. After 4h, the mixture was cool down to rt and filtered on celite. The celite pad was washed with MeOH. The solution was then concentrated to dryness and the residue was purified by flash chromatography with Ethyl Acetate/MeOH (100 to 80-20) to afford a beige solid, (137 mg, 73.9 % yield, purity by HPLC : 95.3 %, LC tR = 2.01 min, MS (ESI ⁺ ): m/z = 511 [M + H] ⁺ ,MS (ESP): m/z = 509 [M + H]’).

In a round-bottom flask, methyl 2-[4-[3-(2,4-dimethylphenyl)sulfonyl-5-oxo-4H- triazolo[l,5-a]quinazolin-8-yl]piperazin-l-yl]acetate (159 mg, 0.311 mmol) was dissolved in a solution of THF-H2O (50:50, 50 ml) then LiOH monohydrate (52.3 mg, 1.25 mmol, 4 eq) was added. The mixture was stirred for 2 h at 0°C then at RT overnight. After this time, THF was removed under reduced pressure, then the mixture was brought to 0°C and HC1 IN was added dropwise until precipitation. The crude solid was filtered and washed with cold EtOH then dried under vacuum overnight. Obtained 140 mg of a white solid, 90% yield.

Alternatively, the crude solid was filtered and triturated with CHCh then dried under vacuum overnight. Obtained 148 mg of a white solid, 95% yield, purity by HPLC: 100 %, LC TR = 1.60 min, MS (ESI ⁺ ): m/z = 497.2 [M + H] ⁺ , MS (EST): m/z = 495.2 [M + H]'.

Intermediates II-2 to II- 11 were prepared following the same procedures as described for Intermediate II- 1 starting from the corresponding raw materials.

Table 1 : Intermediates II-2 to II- 11 General Reaction Conditions for Amide Formation (Using II- 1 as an Illustration).

A suspension of l-(3-dimethylaminopropyl)-3 -ethylcarbodiimide hydrochloride (1.1 eq.) in anhydrous DMF (IM) and DCM (0.5M) was treated with Intermediate II-l then with 1- hydroxybenzotriazole hydrate (1.1 eq.). After the suspension had cleared, the appropriate amine R ⁸¹ R ⁸² NH (1.5 eq.) in dichloromethane (2.5M) was added. The mixture was stirred at room temperature for 18h, then concentrated in vacuo to remove DMF. If needed, the residue was purified by chromatography in appropriate eluent to give the desired product as a solid or tested without further workup.

Alternatively, a solution of 2-[4-[3-(2,4-dimethylphenyl)sulfonyl-5-oxo-4H-triazolo[l,5- a]quinazolin-8-yl]piperazin-l-yl]acetic acid (Intermediate II-l, 2.5 mg, 0.005 mmol) in NMP (0.75 mL) was added to a matrix tube with TMP (0,004 mL, 0.025 mmol, 5 eq) and T3P (0.003 mL, 0.05 mmol, 1 eq). After Ih the appropriate amine R ⁸¹ R ⁸² NH (0.005 mmol, 1 eq) and T3P (0.003 mL, 0.005 mmol, 1 eq) were added to the mixture which was stirred at RT for 16h (using BioShake IQ at 1800 rpm). The crude product was then concentrated to dryness (using Genevac EZ-2 Plus, medium boiling point, 65°C, 7h). The compounds were tested without further purification. Alternative methods of acid activation known in the art can be used as well.

General Reaction Conditions for -Boc Protecting Group Removal

When the amine R ⁸¹ R ⁸² NH contains an additional nitrogen protected by a -Boc group, the following general procedures were used to generate the free amine:

A 50 ml flask was charged with the Boc-protected derivative (1 eq) in dry dioxane (0.35 M), then HC1 4 N in dioxane (15 eq) was added dropwise and the mixture stirred at 80°C. The heating was stopped when the reaction was judged completed by LC-MS then the precipitate was filtered and triturated in Et2O to afford the desired product.

Alternatively, TFA was used: a 50 ml flask was charged with the Boc-protected derivative (1 eq) in dry DCM (1.7 M), then TFA (15 eq) was added dropwise, and the mixture stirred at 40°C. The heating was stopped when the reaction was judged completed by LC-MS, then the mixture was concentrated under reduced pressure and triturated in Et2O to afford the desired product. Alternatively, a matrix tube charged with the Boc-protected derivative was dissolved in dry DCM (0.02 M), then TFA (250 eq) was added dropwise, and the mixture stirred at 40°C (using BioShake IQ at 1800 rpm). The heating was stopped when the reaction was judged completed by LC-MS, then the mixture was concentrated to dryness (using Genevac EZ-2 Plus, medium boiling point, 65°C, 7h) and left under vacuum for 4 days to afford the desired product.

Alternatively, a matrix tube charged with the Boc-protected derivative was dissolved in NMP (0.02 M), then HC1 aq. (37%) was added dropwise, and the mixture was stirred at 80°C. The heating was stopped when the reaction was judged completed by LC-MS, then the mixture was concentrated to dryness (using Genevac EZ-2 Plus, medium boiling point, 65°C, 7h) and left under vacuum for 4 days to afford the desired product.

Example 1: 3-(2.4-dimethylphenyl)sulfonyl-8-[4-(2-oxo-2-pyrrolidin-l-yl -ethyl)piperazin-l- yl |-4H-triazolo| 1.5-a|quinazolin-5-one (Compound I )

Compound 1 was obtained following the general protocol of amide formation from Intermediate II- 1 as described above using pyrrolidine as the amine partner. The title product was isolated as a beige solid (100% purity by HPLC, method A2); or

A round-bottom flask was charged with Intermediate 1-1 (30 mg, 0.0692 mmol, 1 eq), 2- piperazin-l-yl-l-pyrrolidin-l-yl-ethanone (27.3 mg, 0.138 mmol, 2 eq), Pd2dbas (2 mg, 0.00346 mmol, 5 mol%), CS2CO3 (34 mg, 0.104 mmol, 1.5 eq) and RuPhos (3.23 mg, 0.00692 mmol, 10mol%) then DMF (0.035 M) was added. The mixture was degassed by Ar flush then equipped with a reflux condenser and stirred in a preheated heating block at 120°C overnight. After this time the mixture was cool down to rt, the mixture was diluted with MeOH and filtered on celite and concentrated under reduced pressure. The crude product was purified by flash chromatography with 100% DCM until DCM-MeOH (95-5) to afford 13 mg of the title product as a beige solid (34% yield, 100% purity by HPLC, Method A2). LC TR = 1.87, [ES+ MS] m/z 550.3 (MH+). Example 2: 3-(2.4-dimethylphenyl)sulfonyl-8-[4-(2-morpholino-2-oxo-ethy l)piperazin-l-yl]-

4H-triazolo[1.5-a]quinazolin-5-one (Compound 2)

Compound 2 was obtained following the same procedures as described for Compound 1 starting from 2-morpholino-l -piperazin- 1 -yl-ethanone. The title product was isolated as a white solid (100% purity by HPLC, method A2). LC TR = 1.82, [ES+ MS] m/z 566.2 (MH+)

Compounds 3 to 175 of the present invention were prepared from the corresponding Intermediate as indicated in the Table 2 below, following the appropriate general protocol(s) described above. Table 2: Exemplary Compounds of the Invention

Example 3: AgrA Reporter Assay

The 5. aureus USA300 strain UAMS-1625 was transformed with a plasmid expressing the reporter gene lacZ under the control of the P3 promoter, which is regulated by AgrA. The resulting reporter strain was used to measure AgrA inhibitory activity as follows. From an overnight culture plate, cells were resuspended in 0.9% (w/v) saline solution and bacterial inoculum was prepared in LB medium with 5xl0 ⁵ CFU/ml. Compounds were two-fold serially diluted in LB and 50 pL of this lOx concentrated samples were added into the 96-deep well plate. 500 pL of bacterial suspension was finally added to the compounds or DMSO control. Plates were covered and incubated with shaking at 37 °C. After 18 hours incubation, 100 pL was transferred to a new clear 96-well plate and the ODsoo was measured using a TECAN Infinite F200 microplate reader to control for growth inhibition. Another 100 pL was transferred to a new black 96-well plate containing 30 pL of lysis buffer (20 mM Tris/HCl, 3 mM MgCh, 0.5 % Tween 20 (v/v), 0.5 % NP-40 (v/v)) and the plate was incubated with shaking at 37 °C for 60 min. 20 pL of the substrate MUG (4-Methylumbelliferyl P-D-galactopyranoside) at 0.25 mg/ml was added and the fluorescence (excitation: 360 nm, emission: 450 nm) was measured with a TECAN Infinite F200 microplate reader after 3 hours incubation at 37 °C with shaking. The ICso was determined using GraphPad PRISM after normalization to DMSO control.

IC50 in the presence of human serum albumin (HSA, 40mg/mL) was determined by supplementing the LB medium used for inoculum preparation.

The activity of the inventive compounds in the absence of human serum albumin and in the presence of human serum albumin is given in Table 3, below. The corresponding data for Compounds 45 of WO 2020/109350 under the same conditions is additionally provided.

Compound 45 of WO 2020/109350

Table 3: Reporter Assay Results

IC50 (without HSA): +++: IC50 < Ipg/mL, ++: Ipg/mL < IC50 < 5pg/mL

IC50 (with HSA): +++: IC50 < lOpg/mL, ++: lOpg/mL < IC50 < 50pg/mL, +: IC50 > 50pg/mL

Ratio IC50 (in presence of HSA) / IC50 (in absence of HSA): +++: ratio < 25; ++: 25 < ratio < 125; +: ratio > 125