polyamides

Abstract

The present disclosure provides novel polyamide compounds that are useful as binders of DNA, mitochondrial DNA in particular, and in targeted suppression of gene expression. Also provided are novel polyamide compounds comprising a mitochondrial delivery agent or a fluorescent label as well as mitochondria-targeting capsules comprising a novel polyamide compound. Pharmaceutical compositions comprising the compounds or mitochondria-targeting capsules of the disclosure are also provided, as well as medical uses and methods of treatment including the compounds, mitochondria-targeting capsules and pharmaceutical compositions.

Description

[0001] POLYAMIDES

[0002] FIELD

[0003] The present disclosure provides novel polyamide compounds that are useful as binders of DNA, mitochondrial DNA in particular, and in targeted suppression of gene expression. Also provided are novel polyamide compounds comprising a mitochondrial delivery agent or a fluorescent label as well as mitochondria-targeting capsules comprising a novel polyamide compound. Pharmaceutical compositions comprising the compounds or mitochondria-targeting capsules of the disclosure are also provided, as well as medical uses and methods of treatment including the compounds, mitochondria- targeting capsules and pharmaceutical compositions.

[0004] BACKGROUND

[0005] Mitochondria supply 90% of the energy needed by a cell by generating adenosine triphosphate through oxidative phosphorylation of adenosine diphosphate. Mitochondria also play a role in regulating apoptosis of a cell. Thus, mitochondrial dysfunction can lead to problems in all tissues, particularly in tissues that have a high energy demand, such as the central nervous system, and skeletal and cardiac muscles.

[0006] Mitochondrial deoxyribonucleic acids (mtDNAs) are double stranded, circular deoxyribonucleic acids (DNAs) and comprise 37 genes encoding 13 essential subunits of the oxidative phosphorylation system. Mitochondrial dysfunction occurs where one or more of the genes within mtDNA has one or more mutations or defects resulting in the gene being unable to function as it usually would. Often, both mutated or defective and healthy mtDNA co-exist within the same cells, and symptoms of mitochondrial dysfunction arise when the amount of mutated or defective mtDNA increases to a level where oxidative phosphorylation function of the mitochondria falls below a certain threshold. Preventing this can be achieved through the use of N-methylpyrrole-N- methylimidazole polyamides (PIPs), which are known to bind to specific DNA sequences in the minor groove of double-stranded DNA (see, for example, Murty and Sugiyama, Biol. Pharm. Bull., 2004, 27(4), 468-74).

[0007] PIPs are synthetic ligands that recognise predetermined DNA sequences with affinities and specificities comparable to many DNA-binding proteins. Known PIPs are hairpin structures that comprise pyrrole and imidazole moieties. The pyrrole moieties favour thymine (T), adenine (A) and cytosine (C) bases, whereas the imidazole moieties favour guanine (G) bases. Antiparallel pairings of imidazole / pyrrole and pyrrole / imidazole target G:C and C:G, respectively, and antiparallel pairings of pyrrol e / pyrrole target A:T or T:A. Aliphatic β-alanine can be substituted for pyrrole and has been used effectively when molecules have more than five consecutive pyrrole or imidazole residues, by adjusting the pitch between amide bonds and the accepting residue of the minor groove. Antiparallel pairings of pyrrole / β-alanine and β-alanine / pyrrole target A:T or T:A, and antiparallel pairings of imidazole / β-alanine and β-alanine / imidazole specify G:C and C:G, respectively (see, for example, Han et al., Nucleic Acids Research, 2012, 40, 22).

[0008] Hidaka et al., in J. Am. Chem. Soc., 2017, 139, 8444-8447, report the conjugation of mitochondria-penetrating peptide (MPP) with PIPs to generate mitochondria-specific PIPs (termed MITO-PIPs). These were designed to target the known binding site of mitochondrial transcription factor A (TFAM) in the L-strand promoter (LSP) of mDNA, which would result in a reduction in the expression of a downstream gene. In this case, the expression of mitochondrially encoded NADH dehydrogenase 6 was reduced by binding of the MITO-PIPs.

[0009] In US 2015 / 0191571 A1 (KNC Laboratories CO. LTD.), specific polyamide compounds are described which bind to healthy mtDNA and increase replication of healthy mtDNA over A3243G mutant mtDNA, thereby reducing the amount of A3243G mutant mtDNA and increasing the amount of healthy mtDNA in cells.

[0010] Pyrrolidine monomers have previously been incorporated into polyamides, but not in hairpin polyamides employing the Dervan pairing rules. Pyrrolidine use was first reported by C. Woods et al., in Bioorganic & Medicinal Chemistry Letters, 12, 18, 2002, 2647- 2650, to produce single-strand polyamide (ss-PA), also known as linear polyamides, analogues of distamycin, where all three pyrrole groups were replaced by pyrrolidine (or alternatively cyclopentane or tetrahydrofuran). However, all resulting linear polyamides had greatly reduced DNA binding relative to distamycin (measured using a fluorescent indicator displacement (FID) assay).

[0011] In two subsequent papers by Yang et al (C. Lin, Y. Yang and C. Ong, J. Chin. Chem. Soc. 2012, 59: 436-442 and Y. Yang et al. Proteome Sci, 11 , 23 (2013) only one pyrrolidine group of a linear polyamide distamycin-type analogue was replaced with a pyrrolidine (containing a BOC sidechain on the heterocyclic nitrogen). In both papers, a poor DNA binding affinity was measured by assessing changes in the melting temperature of calf-thymus DNA. There was no indication of sequence selectivity.

[0012] A cyclopentyl (Cyp) moiety is reported by H. Zhang et al., in ChemBioChem. 2012, 13, 1366-1374, to have been incorporated into a hairpin polyamide, replacing a pyrrole moiety over an A nucleotide and paired with a pyrrole (Py) moiety over the T nucelotide. A reduction in affinity of Cyp / Py pairing relative to Py / Py pairing was reported.

[0013] Floreancig et al., in “Recognition of the Minor Groove of DNA by Hairpin Polyamides Containing a-Substituted-β-Amino Acids”, J. Am. Chem. Soc., 2000, 6342-6350, 122, describe incorporation of β-alanine (β-) into hairpin polyamides composed of N- methylpyrrole (Py) and N-methylimidazole (Im) amino acids and report that this incorporation is required for selective binding to DNA sequences longer than seven base pairs. It is also reported that DNA binding in the minor groove is sensitive to the stereochemistry, steric bulk, and electronics of the substituent at the a-position of β- amino acids in hairpin polyamides containing β / β pairs.

[0014] In WO 98 / 37067 A1 , polyamides for binding to specific nucleotide sequences of double stranded DNA as well as methods for designing and synthesizing polyamide DNA binding ligands for specific nucleotide sequences are described.

[0015] Supekova et al., in “Genomic Effects of Polyamide / DNA Interactions on mRNA Expression”, 2002, 821-827, 9, describe the biological activity of a hairpin polyamide that inhibits binding of the minor-groove transcription factor LEF-1 , constitutively expressed in colon cancers.

[0016] There is a need in the art to provide alternative polyamides that bind to DNA, such as mtDNA, and that preferably bind in a sequence selective manner. There is also a need in the art to provide alternative polyamides capable of binding to DNA or of modulating DNA activity, such as binding to or modulating mtDNA activity, and having improved physical properties, such as greater solubility in bio-compatible solvents, and / or improved pharmacokinetics. SUMMARY

[0017] The present disclosure is based on the inventors having found that polyamides of formula (I) are surprisingly selective for target DNA sequences and / or have surprisingly beneficial physical and / or pharmacokinetic properties.

[0018] Viewed from a first aspect, therefore, the disclosure provides a compound of formula (I): wherein:

[0019] A is a 5-membered heterocycle optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl; each B1to B7is independently a 5- or 6-membered heterocycle, pentanone, benzene, 5- or 6-membered aliphatic carbocycle or ethan-1 ,2-di-yl, wherein: the 5- or 6- membered heterocycle, pentanone, benzene and 5- or 6-membered aliphatic carbocycle are optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-ehaloalkyl, halo, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl-(5- membered heterocyclyl), C1-4alkyl(C1-4alkoxy)pC1-4alkoxy, C1-4alkylcarboxyl, C1-6alkyl CO2( C1-4alkyl), carboxyl, CO2(C1-4alkyl), CO(C1-4alkyl), C1-4alkylCO(C1-4alkyl), oxo,C2-6alkynyl, C2-6haloalkynyl, amino, hydroxy, C1-6alkyldiC1-4alkylamino, C1-4alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC3-6azacycloalkyl, C1-4alkylmorpholinyl, C1-4alkylCs- ehaloazacycloalkyl, phenyl, 5-membered heterocyclyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2; and the ethan-1 , 2-di-yl is optionally substituted with any one or more selected from hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy wherein the C1-4alkyl substituent may combine with the -NH- group adjacent to the ethan-1 , 2-di-yl to form a C3-5azacycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; each C1and C2is independently selected from a 5- or 6-membered N-heterocycle and ethan-1 ,2-di-yl, wherein the 5- or 6-membered N-heterocycle is optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl-(5-membered heterocyclyl), C1-4alkyl(C1-4alkoxy)pC1-4alkoxy, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), carboxyl, CO2(C1-4alkyl), oxo, C2-6alkynyl, C2-6haloalkynyl, amino, hydroxy, C1-4alkyldiC1-4alkylamino, C1-6alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC3-6azacycloalkyl, C1-4alkylmorpholinyl, C1-4alkylC3-6haloazacycloalkyl, phenyl, 5-membered heterocyclyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2; and the ethan-1 , 2-di-yl is optionally substituted with any one or more selected from hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy; p is 0 to 5;

[0020] D is selected from C3-6azacycloalkyl and aminopropyl, wherein the azacycloalkyl is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy and the aminopropyl is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy, wherein two C1-4alkyl substituents may combine to form a C2-6cycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; and

[0021] R is selected from H, C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl)2, C1-4alkylC(O)NHC1-6alkylN(C1-6alkyl)H and C1-6alkylC(O)NH C1-6alkylNH2.

[0022] The compounds of the first aspect may be capable of penetrating a cell and entering the mitochondria within the cell without further modification. Alternatively, the compounds of the first aspect may be modified in such a way to enhance cell and / or mitochondria penetration. For example, the compounds of the first aspect may be modified so that one or more mitochondrial delivery agents is / are conjugated to the compounds at one or more suitable positions. Suitable positions are those that allow the mitochondrial delivery agent to enhance penetration of the compound into mitochondria and also allow the compound to bind to its target DNA sequence.

[0023] In order to assess penetration of the compounds of the disclosure into a cell and / or mitochondria, the compounds of the first aspect may be modified so that one or more detectable tags is / are conjugated at one or more suitable positions. Suitable positions are those that allow the detectable tag to be detected on entry of the compound into a cell and / or a mitochondria and also allow the compound to bind to its target DNA sequence.

[0024] Thus, viewed from a second aspect, there is provided a compound as defined in the first aspect, modified such that the compound is conjugated to one or more mitochondrial delivery agents and / or detectable tags. For example, the compound of the second aspect may be a compound of formula (1), (2), (3) or (4):

[0025] wherein:

[0026] A, B1to B7, C1, C2, D and R are as defined in the first aspect; R’ is selected from C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl) and C1-6alkylC(O)NHC1-6alkylNH, R2’ is selected from NHC(O)C1-6alkylNH, C(O)NHC1-6alkylNH, NHC(O)C1-6alkylC(O) and C(O)NHC1-ealkylC(O) and

[0027] M is a mitochondrial delivery agent or a detectable tag.

[0028] Alternatively, the compounds of the first aspect may be modified to enhance cell and / or mitochondria penetration by encapsulating the compound within a mitochondria targeting capsule capable of delivering the compound into the mitochondria. For example, the capsule may be capable of fusing with the outer membranes of mitochondria, allowing for delivery of the contents of the capsule (including the compound of the disclosure) into the mitochondria.

[0029] Thus, viewed from a third aspect, there is provided a mitochondria-targeting capsule comprising a compound of the first and / or second aspect.

[0030] As described above, mitochondrial dysfunction occurs where one or more of the genes within mtDNA have (or harbour) one or more mutations or defects resulting in the gene being unable to function as it usually would (or should). Symptoms of mitochondrial dysfunction may arise when the amount of mutated or defective mtDNA increases to a level where the oxidative phosphorylation function of the mitochondria falls below a certain threshold. The compounds of the disclosure are able to bind to specific mtDNA sequences in order to inhibit replication or expression of mutated or defective mtDNA, or to enhance or promote replication or expression of healthy mtDNA over mutated or defective mtDNA. Accordingly, the compounds and capsules of the disclosure are useful in medicine, as medicaments, as pharmaceuticals. By way of example, any of the compounds of this disclosure may be for use in the treatment and / or prevention of diseases and / or conditions caused or contributed to by mutated or defective mtDNA and / or mitochondrial dysfunction.

[0031] Thus, viewed from a fourth aspect, there is provided a pharmaceutical composition comprising one or more compounds of the first and / or second aspects of the disclosure and / or a mitochondria-targeting capsule of the third aspect of the disclosure and a pharmaceutically acceptable excipient.

[0032] Viewed from a fifth aspect, there is provided a compound of the first or second aspect of the disclosure, a mitochondria-targeting capsule of the third aspect of the disclosure, or pharmaceutical composition of the fourth aspect of the disclosure for use in a method of treatment.

[0033] Viewed from a sixth aspect, there is provided a compound of the first or second aspect of the disclosure, a mitochondria-targeting capsule of the third aspect of the disclosure, or pharmaceutical composition of the fourth aspect of the disclosure for use in a method of treating or preventing mitochondrial dysfunction or a disease or condition associated therewith.

[0034] Viewed from a seventh aspect, there is provided a method of treating or preventing mitochondrial dysfunction or a disease or condition associated therewith in a subject comprising administering a therapeutically effective amount of a compound of the first or second aspect of the disclosure, a mitochondria-targeting capsule of the third aspect of the disclosure, or pharmaceutical composition of the fourth aspect of the disclosure, to the subject.

[0035] Viewed from an eighth aspect, there is provided a compound of the first or second aspect of the disclosure, a mitochondria-targeting capsule of the third aspect of the disclosure, or pharmaceutical composition of the fourth aspect of the disclosure for use in medicine or as a medicament. Particular isolatable intermediates used to synthesise the compounds of the first or second aspects of the invention are novel and have been found by the inventors to be particularly useful as components of the compounds of the first or second aspects of the invention. For example, particular intermediates, when included in the compounds of the first or second aspects, have been found to give rise to favourable properties, such as increased solubility and / or selectivity for binding to a particular sequence of mtDNA.

[0036] Therefore, viewed from a ninth aspect, there is provided a compound selected from formulae (i), (ii) and (iii): wherein:

[0037] P is an amine protecting group;

[0038] LG is a leaving group; r1is selected from C1-4alkylC(O)OC1-4alkyl, C1-4alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy, C1-4alkylcyano and C1-4alkylO-P1, wherein P1is an alcohol protecting group; r2is selected from C1-4alkyl and C1-4haloalkyl; r3is selected from C1-4alkyl, halo and C1-4haloalkyl; r4is selected from C1-4haloalkyl, C1-4alkylO-P1and formula (iiia):

[0039] P (iiia); s1and s2are each independently selected from C1-4alkyl and C1-4haloalkyl; and ns1 and ns2 are each independently selected from 0 to 2.

[0040] Viewed from a tenth aspect, there is provided use of a compound of the ninth aspect in the synthesis of polyamides. DEFINITIONS

[0041] In the discussion that follows, reference is made to a number of terms, which are to be understood to have the meanings provided below, unless a context indicates to the contrary. The nomenclature used herein for defining compounds, in particular the compounds described herein, is intended to be in accordance with the rules of the International Union of Pure and Applied Chemistry (IUPAC) for chemical compounds, specifically the “IUPAC Compendium of Chemical Terminology (Gold Book)” (see A. D. Jenkins et al., Pure & Appl. Chem., 68, 2287-2311 (1996)). For the avoidance of doubt, if an IUPAC rule is contrary to a definition provided herein, the definition herein is to prevail.

[0042] The term “comprising” or variants thereof is to be understood herein to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

[0043] The term “consisting” or variants thereof is to be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, and the exclusion of any other element, integer or step or group of elements, integers or steps.

[0044] The term “about” herein, when qualifying a number or value, is used to refer to values that lie within ± 5% of the value specified. For example, where a capsule is defined as having a diameter of about 50 to about 900 nm, diameters of 47.5 to 945 nm are included.

[0045] The term “heterocycle” refers to monocyclic rings having within the ring at least one heteroatom, for example, a heteroatom selected from N, S and O. The ring may contain one or more double bonds and may be aliphatic or aromatic. The ring may be 5- or 6- membered. Any N heteroatom present in the heterocycle may be substituted. Representative examples of heterocycles include, but are not limited to, thiophene, thiazole, pyrrole, imidazole, pyrazole, pyrrolidine, pyrroline, pyrazolidine, imidazolidine, pyrazoline, oxazole, isoxazole, isothiazole, furan, furazan, tetrahydrothiophene, tetra hydrofuran, piperidine, pyridine, piperazine, pyridazine, pyrimidine, pyrazine, tetrahydropyran, 1 ,4-dioxane, 1 ,4-dioxine, thiane, thiopyran, 1 ,4-dithiane, morpholine, oxazine, thiazine and thiomorpholine. As used herein, “optionally substituted heterocycle” refers to a heterocycle as defined herein which comprises one or more substituents on the heterocyclic ring. For the avoidance of doubt, the term “heterocyclyl” refers to a monovalent heterocycle formed by the removal of a hydrogen atom from any carbon atom.

[0046] The term “heteroaromatic ring” refers to compounds derived from aromatic rings, or arenes, by replacement of one or more methine (-C=) and / or vinylene (-CH=CH-) groups by trivalent or divalent heteroatoms, respectively, in such a way as to maintain the continuous TT-electron system characteristic of aromatic systems and a number of out-of-plane TT-electrons corresponding to the Huckel rule. The Huckel rule is often used in the art to assess aromatic character; monocyclic planar (or almost planar) systems of trigonally (or sometimes digonally) hybridised atoms that contain (4n+2) TT-electrons (where n is a non-negative integer) will exhibit aromatic character. Heteroaromatic rings may be 5- or 6-membered. Examples of 5- or 6-membered aromatic rings include pyridine, thiophene, pyrrole, furan, imidazole, oxazole, thiazole, pyrazole, isoxazole and isothiazole.

[0047] Heteroaryl groups are univalent groups derived from heteroaromatic rings by removal of a hydrogen atom from any carbon atom.

[0048] The term “carbocycle” refers to monocyclic rings having only carbon atoms within the ring. The ring may contain one or more double bonds and is aliphatic. The ring may be 5- or 6-membered. Representative examples of carbocycles include, but are not limited to, cyclopentane, cyclopentadiene and cyclohexane. As used herein, “optionally substituted carbocycle” refers to a carbocycle as defined herein which comprises one or more substituents on the cyclic ring. For the avoidance of doubt, the term “carbocyclyl” refers to a monovalent cyclic ring formed by the removal of a hydrogen atom from any carbon atom.

[0049] The term “cycloalkane” refers to saturated monocyclic hydrocarbons. Cycloalkanes may be 5- or 6-membered, i.e. may be cyclopentane or cyclohexane.

[0050] The term “azacycloalkane” refers to compounds derived from cycloalkanes by replacement of one or more methylene (-CH2-) groups by a nitrogen atom. As used herein, “optionally substituted azacycloalkane” refers to an azacycloalkane as defined herein which comprises one or more substituents on the ring, connected at either a carbon or nitrogen ring atom. For the avoidance of doubt, the term “azacycloalkyl” refers to a monovalent azacycloalkane formed by the removal of a hydrogen atom from any carbon atom.

[0051] The term “alkyl” is well known in the art and defines univalent groups derived from alkanes by removal of a hydrogen atom from any carbon atom, wherein the term “alkane” is intended to define acyclic branched or unbranched hydrocarbons having the general formula CnH2n+2, wherein n is an integer >1. Alkyl groups may be C1-4alkyl groups. In some cases, alkyl groups are C1-4alkyl groups. C1-4alkyl refers to any selected from the group consisting of methyl, ethyl, n-propyl, / so-propyl, n-butyl, sec-butyl, / so-butyl and terf-butyl.

[0052] The term “alkylol” defines univalent groups derived from alcohols by removal of a hydrogen atom from a carbon atom, wherein the term “alcohol” is intended to define groups derived from alkanes by the replacement of a hydrogen atom with a hydroxy group. Often, alkylol groups are C1-4alkylol groups or C1-4alkylol groups.

[0053] The term “alkoxy” defines univalent groups derived from alcohols by removal of a hydrogen atom from an OH group. Often, alkoxy groups are C1-4alkoxy groups or C1-4alkoxy groups.

[0054] Halo refers to a halogen radical. Typically, halo refers to any selected from fluoro, bromo, chloro and iodo. In some cases, halo refers to fluoro.

[0055] The term “haloalkyl” is also well known and defines univalent groups derived from alkyl groups by replacement of one or more hydrogen atoms from one or more carbon atoms with a halo group. Haloalkyl groups may comprise one or more different types of halo. For example, one or more independently selected from fluoro, chloro, bromo and iodo. In some cases, the haloalkyl is a fluoroalkyl.

[0056] The term “alkylphenyl” defines univalent groups derived from phenylalkanes by removal of a hydrogen atom from a carbon atom within the alkane. Phenylalkanes are formed by the replacement of a hydrogen atom on a benzene ring with an alkyl group. Alkylphenyl groups are typically C1-4alkylphenyl groups. The term “alkylcycloalkyl” defines univalent groups derived from cycloalkylalkanes by removal of a hydrogen atom from a carbon atom within the alkane. Cycloalkylalkanes are formed by the replacement of a hydrogen atom on a cycloalkane with an alkyl group. Alkylcycloalkyl groups are typically C1-4alkylcycloalkyl, alkylcycloC3-6alkyl, or C1-4alkylcycloC3-6alkyl groups.

[0057] The term “alkylpyridyl” defines univalent groups derived from pyridylalkanes by removal of a hydrogen atom from a carbon atom within the alkane. Pyridylalkanes are formed by the replacement of a hydrogen atom on a carbon ring atom within a pyridine ring with an alkyl group. Alkylpyridyl groups are typically C1-4alkylpyridyl groups.

[0058] The term “C1-6alkylC1-4alkoxy” defines univalent groups derived from alkoxyalkanes by removal of a hydrogen atom from a carbon atom within the alkane. Alkoxyalkanes are formed by the replacement of a hydrogen atom on the hydroxy group of an alcohol with an alkyl group. Alkylalkoxy groups are typically C1-6alkylC1-4alkoxy groups, such as C1-4alkylC1-4alkoxy groups.

[0059] The term “alkylcarboxyl” defines univalent groups derived from carboxyalkanes by removal of a hydrogen atom from a carbon atom within the alkane. The alkylcarboxyl group is typically a C1-4alkylcarboxyl.

[0060] The term “alkylCO2(alkyl)” defines univalent groups derived from alkylcarboxyalkanes by removal of a hydrogen atom from a carbon atom within the alkane. Alkylcarboxyalkanes are formed by the replacement of a hydrogen atom on the hydroxy group of a carboxyalkane with an alkyl group. The alkylCO2(alkyl) is typically a C1-6alkylCO2(alkyl), an alkylCO2(C1-4alkyl), or a C1-6alkylCO2(C1-4alkyl).

[0061] The term “CO(alkyl)” defines univalent groups derived from aldehydes by removal of a hydrogen atom from the C(O)H aldehyde moiety within the aldehyde. “alkylCO(alkyl)” groups are univalent groups derived from ketones by removal of a hydrogen atom from one of the alkyl moieties of the ketone. The CO(alkyl) is typically a CO(C1-4alkyl), or a CO(C1-4alkyl). The alkylCO(alkyl) is typically a C1-6alkylCO(C1-6alkyl) or a C1-4alkylCO(C1-4alkyl). The term “alkynyl” defines univalent groups derived from alkynes by removal of a hydrogen atom from any carbon atom, wherein the term “alkyne” is intended to define acyclic branched or unbranched hydrocarbons having one carbon-carbon triple bond and the general formula CnH2n-2, where n is an integer >2. C2-C4alkynyl refers to any one selected from the group consisting of ethynyl, prop-1-ynyl, prop-2-ynyl, but-1-ynyl, but- 2-ynyl, but-3-ynyl, and 1-methyl-prop-2-ynyl.

[0062] The term “C2-6haloalkynyl” defines univalent groups derived from alkynyl groups by replacement of one or more hydrogen atoms from one or more carbon atoms with a halo group. Haloalkynl groups may comprise one or more different types of halo. For example, one or more independently selected from fluoro, chloro, bromo and iodo. In some cases, the haloalkynl is a fluoroalkynl.

[0063] For the avoidance of doubt, the term “amino” refers to an -NH2group.

[0064] The terms “alkyldialkylamino” and “alkylalkylamino” define univalent groups derived from dialkylaminoalkanes and alkyaminoalkanes respectively by removal of a hydrogen atom from a carbon atom within the alkane. Dialkylaminoalkanes and alkylaminoalkanes are formed by the replacement of a hydrogen atom on the nitrogen atom of an alkylamino or dialkylamino with an alkyl group. The alkyl groups within the alkyldialkylamino and alkylalkylamino typically comprise 1 to 6 or 1 to 4 carbon atoms.

[0065] The term “alkylcyano” defines a univalent group derived from a cyanoalkane by removal of a hydrogen atom from a carbon atom within the alkane. The alkylcyano is typically a C1-4alkylcyano or a C1-4alkylcyano.

[0066] The term “alkylalkoxyalkoxy” defines univalent groups derived from alkoxyalkoxyalkanes by removal of a hydrogen atom from a carbon atom within the alkane. Alkoxyalkoxyalkanes are formed by the replacement of a hydrogen atom on the hydroxy group of an alkoxyalcohol with an alkyl group. The alkyl groups within the alkylalkoxyalkoxy typically comprise 1 to 6 or 1 to 4 carbon atoms.

[0067] The term “alkylazacycloalkyl” defines univalent groups derived from azacycloalkylalkanes by removal of a hydrogen atom from a carbon atom within the alkane. Azacycloalkylalkanes are formed by the replacement of a hydrogen atom on a carbon ring atom within an azacycloalkane with an alkyl group. Alkylazacycloalkyl groups are typically C1-4alkylazacycloalkyl, alkylC3-6azacycloalkyl, or C1-4alkylC3-6azacycloalkyl groups.

[0068] The term “alkylmorpholinyl” defines univalent groups derived from morpholinylalkanes by removal of a hydrogen atom from a carbon atom within the alkane. Morpholinylalkanes are formed by the replacement of a hydrogen atom on a carbon or nitrogen ring atom within a morpholine with an alkyl group. Alkylmorpholinyl groups are typically C1-4alkylmorpholinyl groups.

[0069] The term “alkylhaloazacycloalkyl” defines univalent groups derived from haloazacycloalkylalkanes by removal of a hydrogen atom from a carbon atom within the alkane. Azahalocycloalkylalkanes are formed by the replacement of a hydrogen atom on a carbon ring atom within an azahalocycloalkane with an alkyl group. Alkylhaloazacycloalkyl groups are typically C1-4alkylhaloazacycloalkyl, alkylhaloCs-6azacycloalkyl, or C1-4alkylhaloC3-6azacycloalkyl groups.

[0070] The term “oxo” defines a bivalent oxygen substituent, which connects to another atom, such as carbon, by way of a double bond. For the avoidance of doubt, an oxo substituent replaces two monovalent substituents or one bivalent substituent. Often, an oxo substituent replaces two hydrogen atoms on a carbon atom to form a C=O bond.

[0071] For the avoidance of doubt, a wavy line in a chemical structure bisects the bond linking the moiety shown to the rest of the compound.

[0072] The term “stereoisomer” is used herein to refer to isomers that possess identical molecular formulae and sequence of bonded atoms, but which differ in the arrangement of their atoms in space.

[0073] The term “enantiomer” defines one of a pair of molecular entities that are mirror images of each other and non-superimposable, i.e. cannot be brought into coincidence by translation and rigid rotation transformations. Enantiomers are chiral molecules, i.e. are distinguishable from their mirror image. The term “racemic” is used herein to pertain to a racemate. A racemate defines a substantially equimolar mixture of a pair of enantiomers, which typically comprises a pair of enantiomers in a ratio of about 1 :1.

[0074] The term “diastereoisomers” (also known as diastereomers) defines stereoisomers that are not related as mirror images.

[0075] The term “solvate” is used herein to refer to a complex comprising a solute, such as a compound or salt of the compound, and a solvent. If the solvent is water, the solvate may be termed a hydrate, for example a mono-hydrate, di-hydrate, tri-hydrate etc., depending on the number of water molecules present per molecule of substrate.

[0076] The term “isotope” is used herein to define a variant of a particular chemical element, in which the nucleus necessarily has the same atomic number but has a different mass number owing to it possessing a different number of neutrons.

[0077] The term "treatment" or “therapy” defines the therapeutic treatment of a human or nonhuman animal, in order to impede or reduce or halt the rate of the progress of a disease or condition, or to ameliorate or cure the disease or condition. Prophylaxis of the disease or condition as a result of treatment is also included. References to prophylaxis are intended herein not to require complete prevention of a disease or condition: its development may instead be hindered through treatment in accordance with the disclosure. Typically, treatment is not prophylactic, and the compound or composition is administered to a patient having a diagnosed or suspected disease or condition. By an "effective amount" herein defines an amount of the compound or composition of the disclosurethat is sufficient to impede the noted disease or condition and thus produce the desired therapeutic or inhibitory effect.

[0078] As used herein, the term “patient” or “subject” is used to describe an animal, such as a mammal (e.g. a human or a domesticated animal), to whom treatment, including prophylactic treatment, with the compounds and / or compositions according to the present disclosure is provided. For treatment of those infections, conditions or disease states which are specific to a specific animal such as a human patient, the term patient refers to that specific animal, including a domesticated animal such as a dog or cat or a farm animal such as a horse, cow, sheep, etc. In general, in the present disclosure, the term patient refers to a human patient unless otherwise stated or implied from the context of the use of the term.

[0079] The term “liposome” is well known in the art and refers to a self-assembling spherical artificial vesicle having at least one lipid bilayer, and which encloses an aqueous space. Typically, liposomes are composed of one or more selected from phospholipids, such as sphingolipids (e.g. sphingomyelin) or phosphatidylcholine, lipopetides, and cholesterol. See Weissig et al., Journal of Liposome Research, 2006, 16, 3, 249-264, for a review of liposomes and liposome-like vesicles for the delivery of actives to mitochondria.

[0080] The term “DQAsome” is also well known in the art and refers to mitochondriotropic cationic bolalipid-based vesicles. Typically, DQAsomes comprise dequalinium. See Weissig et al., Methods Mol. Biol., 2021 , 2275, 13-25, for an update on DQAsomes as mitochondria-targeted pharmaceutical nanocarriers.

[0081] The term “polymer nanoparticle” refers to nano-sized spherical particles comprising a polymeric shell capable of encapsulating actives. Typically, the polymeric shell comprises one or more selected from polysaccharides, chitosan, alginate, dextran, proteins, polyesters, acrylate and methacrylates, and poly(ethylene glycol). See Deng et al., Nanomaterials (Basel), 2020, 10(5), 847, for a review of the use of polymeric nanocapsules in drug delivery.

[0082] The term “inorganic nanoparticle” refers to nano-sized spherical particles comprising a polymeric shell composed of inorganic materials (i.e. materials not containing carbon), capable of encapsulating actives. Typically, the polymeric shell of inorganic nanoparticles comprises hydroxyapatite. See Xu et al, Materials Today Chemistry, 2019, 12, 240-260 for an overview of recent progress on mitochondrial targeted cancer therapy based on inorganic nanoparticles.

[0083] DETAILED DESCRIPTION

[0084] As described above, the present disclosure is based on the inventors having found that polyamides of formula (I), comprising an optionally substituted C3-6azacycloalkyl or a substituted aminopropyl at the hairpin loop, have unexpectedly improved selectivity and / or physical and / or pharmacokinetic properties. Formula (I) comprises A, which is a 5-membered heterocycle optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl. The optionally substituted 5-membered heterocycle comprises one or more heteroatoms, typically one or two heteroatoms. The heteroatoms may be one or more selected from nitrogen, oxygen, and sulfur atoms. The optionally substituted 5-membered heterocycle may be aromatic or aliphatic. For example, the optionally substituted 5-membered heterocycle may be selected from the group consisting of pyrrole, imidazole, pyrazole, pyrrolidine, pyrroline, pyrazolidine, imidazolidine, pyrazoline, oxazole, isoxazole, thiazole, isothiazole, thiophene, furan, furazan, tetrahydrothiophene and tetrahydrofuran, each of which is optionally substituted.

[0085] In some embodiments, A is an optionally substituted N-heteroaryl, e.g. selected from the group consisting of pyrrole, imidazole, pyrazole, oxazole, isoxazole, thiazole, isothiazole, furan and furazan, each of which is optionally substituted. The N-heteroaryl may comprise 1 or 2 nitrogen atoms. In some embodiments, A is an optionally substituted pyrrole or imidazole.

[0086] A is optionally substituted with one or more substituents selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl. The 5-membered heterocyclyl may comprise one or more heteroatoms, typically one or two heteroatoms. The heteroatoms may be one or more selected from nitrogen, oxygen, and sulfur atoms, typically nitrogen and sulphur atoms. The 5-membered heterocyclyl may be aromatic or aliphatic. For example, the 5- membered heterocyclyl may be selected from the group consisting of thiophenyl, thiazolyl, pyrrolyl, imidazolyl, pyrazolyl, pyrrolidinyl, pyrrolinyl, pyrazolidinyl, imidazolidinyl, pyrazolinyl, oxazolyl, isoxazolyl, isothiazolyl, furanyl, furazanyl, tetrahydrothiophenyl and tetrahydrofuranyl. In some embodiments, the 5-membered heterocyclyl is a heteroaryl, e.g. selected from the group consisting of thiophenyl, thiazolyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, isothiazolyl, furanyl and furazanyl. The heteroaryl may comprise 1 or 2 heteroatoms selected from nitrogen and sulfur. In some embodiments, 5-membered heterocyclyl is thiophenyl or thiazolyl. In some embodiments, A is optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-2alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-4azacycloalkyl, thiophenyl and thiazolyl.

[0087] In some embodiments, A is of formula (Ila) or (lib): wherein R1to R4are independently H or as defined above with respect to the optional substituents of A.

[0088] In some embodiments, R1and R2of formulae (Ila) and (lib) are each independently selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl; and R3and R4of formula (lib) are each independently selected from hydrogen, halo, C1-4alkyl and C1-4haloalkyl. In some embodiments, R1is selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-2alkylphenyl, C1-2alkyl C5-6cycloalkyl, C2-6azacycloalkyl, thiophenyl and thiazolyl; R2is C1-4alkyl; and R3and R4are each independently selected from H and halo. In specific embodiments, A is of formula (Ila) and R1is C1-4alkyl, such as methyl.

[0089] In some embodiments, A is of formula (Ila) and R1is selected from C2-6alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-6azacycloalkyl, and 5- membered heterocyclyl. In some embodiments, A is of formula (Ila) and R1is selected from C2-6alkyl, C1-4alkylol, C1-4haloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl. In some embodiments, A is of formula (Ila) and R1is selected from C2-6alkyl, C1-4alkylol, C1-4haloalkyl, C1-2alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-4azacycloalkyl, thiophenyl and thiazolyl. In some embodiments, A is of formula (Ila) and R1is selected from C2-6alkyl, C1-4alkylol, C1-4haloalkyl, C3-4azacycloalkyl, thiophenyl and thiazolyl.

[0090] Formula (I) comprises B1to B7. Each B1to B7is independently a 5- or 6-membered heterocycle, pentanone, benzene, 5- or 6-membered aliphatic carbocycle or ethan-1 ,2- di-yl, each of which are optionally substituted. The 5- or 6-membered heterocycle, pentanone, benzene and 5- or 6-membered aliphatic carbocycle are optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl-(5-membered heterocyclyl), C1-4alkyl(C1-4alkoxy)pC1-4alkoxy, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), carboxyl, CO2(C1-4alkyl), CO(C1-4alkyl), C1-4alkylCO(C1-4alkyl), oxo, C2-6alkynyl, C2- ehaloalkynyl, amino, hydroxy, C1-6alkyldiC1-4alkylamino, C1-6alkylC1-4alkylamino, C1-ealkylcyano, C1-4alkylC3-6azacycloalkyl, C1-4alkylmorpholinyl, C1-4alkylC3-ehaloazacycloalkyl, phenyl, 5-membered heterocyclyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2, where p is 0 to 5.

[0091] In some embodiments, each B1to B7is independently a 5- or 6-membered heterocycle, pentanone, benzene or ethan-1 ,2-di-yl, each of which are optionally substituted. The 5- or 6-membered heterocycle, pentanone, benzene and 5- or 6-membered aliphatic carbocycle are optionally substituted with any one or more selected from C1-4alkyl, C1-ealkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl-(5-membered heterocyclyl), C1-4alkyl(C1-4alkoxy)pC1-4alkoxy, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), carboxyl, CO2(C1-4alkyl), oxo, C2-6alkynyl, C2-6haloalkynyl, amino, hydroxy, C1-6alkyldiC1-4alkylamino, C1-6alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylCs-6azacycloalkyl, C1-4alkylmorpholinyl, C1-4alkylC3-6haloazacycloalkyl, phenyl, 5-membered heterocyclyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2, where p is 0 to 5.

[0092] The optionally substituted 5- or 6-membered heterocycle of B1to B7may comprise one or more heteroatoms selected from nitrogen, sulphur and oxygen, and may be selected from the group consisting of pyrrole, pyrazole, furazan, thiophene, thiazole, imidazole, pyrrolidine, tetra hydrofuran, imidazolidine, piperidine, oxazolidine, pyrroline, pyrazolidine, imidazolidine, pyrazoline, oxazole, isoxazole, isothiazole, furan, and tetrahydrothiophene. In some embodiments, the optionally substituted 5- or 6- membered heterocycle is selected from the group consisting of pyrrole, pyrazole, furazan, thiophene, thiazole, imidazole, pyrrolidine, tetra hydrofuran, imidazolidine, piperidine, oxazolidine. In some embodiments, the optionally substituted 5- or 6- membered heterocycle is an optionally substituted 5-membered heterocyle. In some embodiments, the 5-membered heterocycle is a heteroarene, e.g. selected from the group consisting of thiophene, thiazole, pyrrole, imidazole, pyrazole, oxazole, isoxazole, isothiazole, furan and furazan. The optionally substituted 5- or 6-membered heterocycle of B1to B7may comprise 1 to 3 heteroatoms selected from nitrogen, sulphur and oxygen. For example, the 5- or 6- membered heterocycle may comprise 1 or 2 heteroatoms selected from nitrogen and sulphur.

[0093] The optionally substituted 5- or 6-membered aliphatic carbocycle of B1to B7may be an optionally substituted 5-membered aliphatic carbocycle, such as optionally substituted cyclopentane or cyclopentadiene.

[0094] In some embodiments, each of B1to B7is independently selected from an optionally substituted pyrrole, pyrazole, furazan, thiophene, pyrrolidine, tetra hydrofuran, imidazolidine, piperidine, and oxazolidine; an optionally substituted pentanone; an optionally substituted benzene; an optionally substituted cyclopentane; and an optionally substituted ethan-1 ,2-di-yl. In some embodiments, each of B1to B7is independently selected from an optionally substituted pyrrole, pyrazole, furazan, thiophene, pyrrolidine, tetra hydrofuran, imidazolidine, piperidine, and oxazolidine; an optionally substituted pentanone; an optionally substituted benzene; and an optionally substituted ethan-1 ,2- di-yl.

[0095] The 5- or 6-membered heterocycle, pentanone, benzene, and 5- or 6-membered aliphatic carbocycle of B1to B7are optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl-(5-membered heterocyclyl), C1-4alkyl(C1-4alkoxy)pC1-4alkoxy, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), carboxyl, CO2(C1-4alkyl), CO(C1-4alkyl), C1-4alkylCO(C1-4alkyl), oxo, C2-6alkynyl, C2-6haloalkynyl, amino, hydroxy, C1-6alkyldiC1-4alkylamino, C1-6alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC3-6azacycloalkyl, C1-4alkylmorpholinyl, C1-4alkylC3-6haloazacycloalkyl, phenyl, 5-membered heterocyclyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2, where p is 0 to 5. The 5- membered heterocyclyl may comprise one or more heteroatoms, typically one or two heteroatoms. The heteroatoms may be one or more selected from nitrogen, oxygen, and sulfur atoms, typically nitrogen and sulphur atoms. The 5-membered heterocyclyl may be aromatic or aliphatic. For example, the 5-membered heterocyclyl may be selected from the group consisting of thiophenyl, thiazolyl, pyrrolyl, imidazolyl, pyrazolyl, pyrrolidinyl, pyrrolinyl, pyrazolidinyl, imidazolidinyl, pyrazolinyl, oxazolyl, isoxazolyl, isothiazolyl, furanyl, furazanyl, tetrahydrothiophenyl and tetrahydrofuranyl. In some embodiments, the 5-membered heterocyclyl is a heteroaryl, e.g. selected from the group consisting of thiophenyl, thiazolyl, pyrrolyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, isothiazolyl, furanyl and furazanyl. The heteroaryl may comprise 1 or 2 heteroatoms selected from nitrogen and sulfur. In some embodiments, the 5-membered heterocyclyl is thiophenyl or thiazolyl.

[0096] In some embodiments, the 5- or 6-membered heterocycle, pentanone, benzene, and 5- or 6-membered aliphatic carbocycle of B1to B7are each independently optionally substituted with one or more substituents selected from C1-4alkyl, halo, C1-4alkylCO2(C1-4alkyl), CO(C1-4alkyl), C1-4alkylcyano, C1-4alkylcarboxyl, and C1-4alkyl(C1-4alkoxy)i-5, such as C1-4alkyl(C1-4alkoxy)2-5, e.g. C1-6alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxy. In some embodiments, the 5- or 6-membered heterocycle, pentanone, benzene, and 5- or 6-membered aliphatic carbocycle, such as the 5- or 6-membered heterocycle, pentanone, and benzene, of B1to B7are each independently optionally substituted with one or more substituents selected from C1-4alkyl, halo, C1-6alkylC1-4alkoxy, C1-6alkylCO2(C1-4alkyl), Ci -ealkylcyano, C1-4alkylcarboxyl, and C1-4alkylC1-4alkoxyC1-4alkoxy.

[0097] In some embodiments, B1to B7is an optionally substituted 5-membered heterocycle, benzene, 5-membered aliphatic carbocycle or ethan-1 ,2-di-yl, such as an optionally substituted 5-membered heterocycle, benzene or ethan-1 , 2-di-yl. In some embodiments, the 5-membered heterocycle and benzene are each independently optionally substituted with one or more substituents selected from C1-4alkyl, halo, C1-6alkylCO2(C1-4alkyl), C1-4alkylcyano, C1-4alkylcarboxyl, and C1-4alkyl(C1-4alkoxy)i-5, such as C1-4alkyl(C1-4alkoxy)2-5, e.g. C1-4alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxy. In some embodiments, the 5- membered heterocycle and benzene are each independently optionally substituted with one or more substituents selected from C1-4alkyl, halo, C1-6alkylCO2(C1-4alkyl), C1-ealkylcyano, C1-4alkylcarboxyl, and C1-4alkylC1-4alkoxyC1-4alkoxy.

[0098] In some embodiments, the optionally substituted 5- or 6-membered aliphatic carbocycle is unsubstituted cyclopentane. The ethan-1 ,2-di-yl of B1to B7is optionally substituted with any one or more selected from hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy, such as methyl or hydroxy. In some embodiments, each of B1to B7is independently selected from formulae (Illa) to

[0099] (Ilin), such as formulae (Illa) to (IIIm): wherein * labels the wavy line that dissects the bond to carbonyl, each X is independently selected from NR9, O and C(O), each of R5to R12are independently H or are as defined above with respect to the optional substituents of B1to B7, n is 0 to 2, n1 is 0 or 1 and n2 is 0 to 3.

[0100] In some embodiments, R5and R7are each independently selected from H, C1-4alkyl, C1-ealkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1- ealkylcarboxyl, C1-4alkylCO2C1-4alkyl, C1-6alkyldiC1-4alkylamino, C1-6alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkyl(C1-4alkoxy)i-5 (such as C1-4alkyl(C1-4alkoxy)2-5, e.g. C1-4alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxy), C1-4alkylC3-6azacycloalkyl, C1-4alkylC1-sheteroaryl, and C1-4alkylCs- ehaloazacycloalkyl. In some embodiments, R5and R7are each independently selected from H, C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylCs- ecycloalkyl, C1-6alkylC1-4alkoxy, C1-4alkylcarboxyl, C1-4alkylCO2C1-4alkyl, C1-4alkyldiC1-4alkylamino, C1-6alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC3-6azacycloalkyl, C1-4alkylC1-3heteroaryl, and C1-4alkylC3-6haloazacycloalkyl.

[0101] In more specific embodiments, R5is selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkylcarboxyl, C1-4alkylCO2C1-4alkyl, C1-4alkyldiC1-4alkylamino, C1-4alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkyl(C1-4alkoxy)i-5 (e.g. C1-4alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylC1-4a I koxy C 1 -4a I koxy C 1 -4a I koxy C 1 -4a I koxy C 1 -4a I koxy) , C 1 -4a I ky I C3-6azacy cl oa I ky I , C 1 -4a I ky I C 1. sheteroaryl, and C1-4alkylC3-6haloazacycloalkyl. In yet more specific embodiments, R5is selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), C1-4alkylcyano, and C1-4alkyl(C1-4alkoxy)i-5 (such as C1-4alkylC1-4alkoxy, C1-4a I ky I C 1 -4a I koxy C 1 -4a I koxy or C 1 -4a I ky I C 1 -4a I koxy C 1 -4a I koxy C 1 -4a I koxy C 1 -4a I koxy C 1. 4alkoxy). In some embodiments, R5is not an alkylcarboxyl. For example, R5may be selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylCs-6cycloalkyl, C1-6alkylCO2C1-4alkyl, C1-4alkyldiC1-4alkylamino, C1-4alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkyl(C1-4alkoxy)i-5 (e.g. C1-4alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxy), C1-4alkylCs-6azacycloalkyl, C1-4alkylC1-3heteroaryl, and C1-4alkylC3-6haloazacycloalkyl.

[0102] In some embodiments, C1-4alkyl(C1-4alkoxy)i-5 is C1-4alkylC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxy.

[0103] In some embodiments, R6is selected from C1-4alkyl, C1-4haloalkyl and halo. In more specific embodiments, R6is C1-4alkyl.

[0104] In some embodiments, R8is selected from H, C1-4alkyl, C1-4haloalkyl, halo, C2-6alkynyl, C2-ehaloalkynyl, amino and phenyl. In more specific embodiments, R8is selected from H, C1-4alkyl, C1-4haloalkyl, halo, C2-6alkynyl, C2-6haloalkynyl, amino and phenyl. In yet more specific embodiments, R8is H.

[0105] In some embodiments, each R9is independently selected from H, C1-4alkyl (such as methyl), C(O)C1-4alkyl (such as C(O)CHs), SO(C1-4alkyl) (such as SOCH3), SO2(C1-4alkyl) (such as SO2CH3) and SO2NH2, such as from H, C1-4alkyl and C(O)C1-4alkyl. In particular embodiments, each R9is H or methyl.

[0106] In some embodiments, each R10is independently selected from C1-4alkyl, C1-4haloalkyl, halo, hydroxy, amino and C1-4alkoxy, such as methyl, trifluoromethyl, difluoromethyl, fluoro, hydroxy, amino and methoxy.

[0107] In some embodiments, each R11and R12is independently selected from H, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, NH2, phenyl and C1-4alkoxy, wherein the C1-4alkyl substituent may combine with the -NH- group adjacent to the ethan-1 ,2-di-yl to form a C3-5azacycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy.

[0108] For the avoidance of doubt, where the C1-4alkyl substituent is described herein to combine with the -NH- group adjacent to the ethan-1 ,2-di-yl to form an optionally substituted C3-5azacycloalkane, the resultant C(O)-NH-B1-7-C(O) structure is of the following formula: wherein S is selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy. In some embodiments, the C3-5azacycloalkane is unsubstituted. In some embodiments, the C3-5azacycloalkane is pyrrolidine.

[0109] In more specific embodiments, R11is H or C1-4alkyl, wherein the C1-4alkyl substituent may combine with the -NH- group adjacent to the ethan-1 ,2-di-yl to form a C3-5azacycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; and R12is selected from H, hydroxy, C1-4alkyl, NH2, and phenyl.

[0110] In yet more specific embodiments, R5and R7are each independently selected from H, C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylCs- ecycloalkyl, C1-4alkylcarboxyl, C1-6alkyldiC1-4alkylamino, C1-6alkylC1-4alkylamino, C1-ealkylcyano, C1-4alkyl(C1-4alkoxy)i-5 (such as C1-4alkyl(C1-4alkoxy)2-5, e.g. C1-4alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxy), C1-4alkylC3-6azacycloalkyl, C1-4alkylC1-3heteroaryl, and C1-4alkylC3-ehaloazacycloalkyl; R6is selected from C1-4alkyl, C1-4haloalkyl and halo; R8is selected from H, C1-4alkyl, C1-4haloalkyl, halo, C2-6alkynyl, C^ehaloalkynyl, amino and phenyl; each R9is independently selected from H, C1-4alkyl, C(O)C1-4alkyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2; each R10is independently selected from C1-4alkyl, C1-4haloalkyl, halo, hydroxy, amino and C1-4alkoxy, and each R11and R12is independently selected from H, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, NH2, phenyl and C1-4alkoxy. In some embodiments, the C1-4alkyl(C1-4alkoxy)i-5 is C1-4alkylC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxy.

[0111] In some embodiments, R6and R7are each independently C1-4alkyl.

[0112] In more specific embodiments, R5is selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-cycloalkyl, C1-4alkylcarboxyl, C1-4alkyldiC1- kylcyano, C1-4alkyl(C1-4alkoxy)i-5 (such as C1-4alkyl(C1-4alkoxy)2-5, e.g. C1-6alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylCi . 4a I koxy C 1 -4a I koxy C 1 -4a I koxy C 1 -4a I koxy C 1 -4a I koxy) , C 1 -4a I ky I C3-6azacy cl oa I ky I , C 1 -4a I ky I C 1. sheteroaryl, and C1-4alkylC3-6haloazacycloalkyl; R6and R7are each independently C1-ealkyl; R8is selected from H, C1-4alkyl, C1-4haloalkyl, halo, C2-6alkynyl, C2-6haloalkynyl, amino and phenyl; each R9is independently selected from H, C1-4alkyl, C(O)C1-4alkyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2; each R10is independently selected from C1-4alkyl, C1-4haloalkyl, halo, hydroxy, amino and C1-4alkoxy; R11is H or C1-4alkyl; and R12is selected from H, hydroxy, C1-4alkyl, NH2, and phenyl. In some embodiments, C1-4alkyl(C1-4alkoxy)i-5 is C1-4alkylC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxy.

[0113] In yet more specific embodiments, R5is selected from C1-4alkyl, C1-4haloalkyl, C1-4alkylpyridyl, C1-4alkylcarboxyl, C1-6alkyldiC1-4alkylamino, C1-4alkyl(C1-4alkoxy)i-5 (such as C1-4alkyl(C1-4alkoxy)2-5, e.g. C1-6alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxy), C1-4alkylC3-6azacycloalkyl and C1-4alkylC1-3heteroaryl; n, n1 and n2 are each 0; R7is C1-4alkyl; and R8is H. In some embodiments, C1-4alkyl(C1-4alkoxy)i-5 is C1-4alkylC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxy.

[0114] In some embodiments, B1, B3, B4and B7are of formula (Illa). In such embodiments, R5of B4and B7may be independently selected from C1-4alkyl, such as methyl. R5of B7may be C1-4alkyl(C1-4alkoxy)i-5 (such as C1-4alkyl(C1-4alkoxy)2-5, e.g. C1-6alkylC1-4alkoxy, C1-4alkylC1-4alkoxy, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), C1-4alkylcyano, and C1-4alkoxy). R5of B3may be selected from C1-4alkyl, C1-6alkylCO2(C1-4alkyl), C1-ealkylcarboxyl, C1-4alkylC3-6azacycloalkyl, such as C1-4alkyl and C1-6alkylCO2(C1-4alkyl). In some embodiments, R5of B3is selected from C1-4alkyl, C1-4alkylCO2(C1-4alkyl), and C1-4alkylcarboxyl.

[0115] In some embodiments, B3is of formula (Illa) and R5of B3is selected from C1-4alkylol, C1-4alkylC1-4alkoxy, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), C1-4alkylcyano, and C1-4alkylC1-4alkoxyC1-4alkoxy, such as C1-4alkylCO2(C1-4alkyl).

[0116] In some embodiments, B2is selected from formulae (Illa) to (Illi) and (Ilin), such as (Illa) to (Illi). In some embodiments, B2is an optionally substituted pyrrolidine, for example B2is selected from formulae (Illg) to (Illi), such as (Illg), where X is NR9.

[0117] In some embodiments, B2is selected from: formula (Illa), wherein R5is selected from C1-4alkylol, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), C1-4alkylcyano, and C1-4alkyl(C1-4alkoxy)i-5 (such as C1-4alkyl(C1-4alkoxy)2-5, e.g. C1-6alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylC1-4a I koxy C 1 -4a I koxy C 1 -4a I koxy C 1 -4a I koxy C 1 -4a I koxy) ; formula (lllc), wherein R7is C1-4alkyl and R8is selected from C1-4alkyl, C1-4haloalkyl and halo; formula (Hid); formula (Illg), wherein X is NR9, R9is selected from H and CO(C1-4alkyl) and each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo; formula (IIIm), wherein one of R11and R12is ethyl and combines with the -NH- group adjacent to the ethan-1 ,2-di-yl to form pyrrolidinyl that is optionally substituted with any one or more selected from C1-4alkyl, halo and C1-4haloalkyl; and formula (Ilin), wherein each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo.

[0118] In some embodiments, where B2is of formula (Illa), R5is selected from C1-4alkylol, C1-4alkylCO2(C1-4alkyl), C1-4alkylcyano, and C1-4alkyl(C1-4alkoxy)i-5 (such as C1-4alkyl(C1-4alkoxy)2-5, e.g. C1-4alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylC1-4aIkoxyC1-4aIkoxyC1-4aIkoxyC1-4aIkoxyC1-4aIkoxy .

[0119] In some embodiments, where B2is selected from formula (IIIm), the resultant C(O)-NH- B2-C(O) is of the following formula:

[0120] In some embodiments, B2is selected from: formula (Illa), wherein R5is selected from C1-4alkylol, C1-4alkylC1-4alkoxy, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), C1-4alkylcyano, and C1-4alkylC1-4alkoxyC1-4alkoxy; formula (lllc), wherein R7is C1-4alkyl and R8is selected from C1-4alkyl, C1-4haloalkyl and halo; formula (Illg), wherein X is NH and each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo; and formula (IIIm), wherein one of R11and R12is ethyl and combines with the -NH- group adjacent to the ethan-1 ,2-di-yl to form pyrrolidinyl that is optionally substituted with any one or more selected from C1-4alkyl, halo and C1-4haloalkyl.

[0121] In some embodiments, B2is selected from formula (Illg), wherein X is NR9, R9is selected from H and CO(C1-4alkyl) and each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo. The inventors have found that such B2groups, whilst likely to improve the physicochemical properties of the compounds, have little or no negative impact on the binding or selectivity of the compounds. Particularly beneficial compounds are those in which B2is of formula (Illa), (lllc), (Illg), (IIIm) and (Ilin), such as (Illg). The inventors have found that such compounds have improved selectivity.

[0122] In some embodiments, B2is of formula (Illg) to (Illi), such as (Illg), (lllh) or (Illi).

[0123] In some embodiments, B5is an optionally substituted pyrrole or an optionally substituted ethan-1 ,2-di-yl.

[0124] In some embodiments, B5is of formula (Illa), (lllc), (Illg) or (IIIm), such as (Illa) or (IIIm). In such embodiments, R5of B5may be selected from C1-4alkyl, C1-4alkylcarboxyl and C1-4alkylC3-6azacycloalkyl. R7and R8of B5may be independently selected from H and C1-4alkyl, for example, R7may be C1-4alkyl and R8may be H. X of B5may be NH and each R10may be independently selected from C1-4alkyl, C1-4haloalkyl and halo. R11and R12of B5may each be independently selected from H and C1-4alkyl.

[0125] In particular embodiments, B5is: of formula (Illg), wherein X is NH and each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo; or of formula (IIIm), wherein R11is C1-4alkyl and R12is H or C1-4alkyl.

[0126] In particular embodiments, B5is: of formula (Illa), wherein R5is selected from C1-4alkyl, C1-4alkylcarboxyl and C1-4alkylC3-6azacycloalkyl, such as C1-4alkyl; or of formula (IIIm), wherein R11is C1-4alkyl and R12is H or C1-4alkyl, or wherein one of R11and R12is ethyl and combines with the -NH- group adjacent to the ethan-1 ,2-di-yl to form pyrrolidinyl that is optionally substituted with any one or more selected from C1-4alkyl, halo and C1-4haloalkyl, or is unsubstituted.

[0127] In particular embodiments, B2is an optionally substituted pyrrolidine and B5is an optionally substituted pyrrole or an optionally substituted ethan-1 ,2-di-yl. For example, in some embodiments, B2is selected from formulae (Illg) to (Illi), such as (Illg), where X is NR9, and B5is selected from formula (Illa) or (IIIm). In some embodiments, B6is selected from formula (Illa), (Illg) and (IIIm), such as formula (Illa). In such embodiments, R5of B6may be C1-4alkyl, such as methyl. Alternatively, R5of B6may be C1-4alkyl(C1-4alkoxy)i-5 (such as C1-4alkyl(C1-4alkoxy)2-5, e.g. C1-4alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy or C1-4alkylC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxyC1-4alkoxy). R11of B6may be H. R12of B6may be selected from H, hydroxy, C1-4alkyl, NH2, and phenyl.

[0128] Formula (I) comprises C1and C2. Each C1and C2is independently selected from a 5- or 6-membered N-heterocycle and ethan-1 ,2-di-yl, each of which is optionally substituted.

[0129] The optionally substituted 5- or 6-membered N-heterocycle of C1and C2may be selected from the group consisting of pyrrole, thiophene, thiazole, imidazole, pyrazole, pyrrolidine, pyrroline, pyrazolidine, imidazolidine, pyrazoline, oxazole, isoxazole, isothiazole, furan, furazan, tetrahydrothiophene, tetra hydrofuran, piperidine, pyridine, piperazine, pyridazine, pyrimidine, pyrazine, tetrahydropyran, 1 ,4-dioxane, 1 ,4-dioxine, thiane, thiopyran, 1 ,4-dithiane, morpholine, oxazine, thiazine and thiomorpholine, each of which is optionally substituted. The N-heterocycle may be aliphatic or aromatic and may comprise 1 or 2 nitrogen atoms. In some specific embodiments, the 5- or 6-membered N-heterocycle of C1and C2is pyrrole or pyrrolidine, each of which is optionally substituted.

[0130] The 5- or 6-membered N-heterocycle of C1and C2is optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl-(5-membered heterocyclyl), C1-4alkyl(C1-4alkoxy)pC1-4alkoxy, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), carboxyl, CO2(C1-4alkyl), oxo, C2-6alkynyl, C2-6haloalkynyl, amino, hydroxy, C1-6alkyldiC1-4alkylamino, C1-4alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC1-4alkoxyC1-4alkoxy, C1-4alkylC3-6azacycloalkyl, C1-4alkylmorpholinyl, C1-4alkylC3-6haloazacycloalkyl, phenyl, 5-membered heterocyclyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2.

[0131] In some embodiments, the 5- or 6-membered N-heterocycle of C1and C2is optionally substituted with one or more substituents selected from C1-4alkyl, halo, C1-4alkylC1-4alkoxy, C1-6alkylCO2(C1-4alkyl), Ci -ealkylcyano, C1-4alkylcarboxyl, and C1-4alkylC1- 4alkoxyC1-4alkoxy. In specific embodiments, the 5- or 6-membered N-heterocycle of C1and C2is optionally substituted with C1-4alkyl, such as methyl.

[0132] The ethan-1 ,2-di-yl of C1and C2is optionally substituted with any one or more selected from hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy, such as methyl or hydroxy. In some embodiments, the ethan-1 ,2-di-yl of C1and C2is unsubstituted.

[0133] In some embodiments, each C1and C2is independently selected from formulae (IVa) to (IVc): wherein * labels the wavy line that dissects the bond to carbonyl and R5, R11and R12are independently H or are as defined above with respect to the optional substituents of C1and C2.

[0134] In specific embodiments, R5of formula (IVa) is C1-4alkyl, such as methyl. In specific embodiments, each R11and R12of formula (IVc) is independently selected from H, hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy. In some embodiments, R11may be H and C2may be of formula (IVb). In yet more specific embodiments, R11of formula (IVc) is H.

[0135] In some embodiments, C2is of formula (IVa) or (IVc), such as (IVc).

[0136] In some embodiments, wherein C1is of formula (IVc), R11is H and R12is independently selected from C1-4alkyl, C1-4haloalkyl and C1-4alkoxy.

[0137] In some embodiments, wherein C1and C2are independently selected from formulae (IVa) and (IVc).

[0138] Formula (I) additionally comprises D, which is selected from C3-6azacycloalkyl and aminopropyl, wherein the C3-6azacycloalkyl is optionally substituted and the aminopropyl is substituted. The C3-6azacycloalkyl, may be selected from piperidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperazinyl, aziridinyl, azetidinyl and diazetidinyl, each of which is optionally substituted. In some embodiments, the C3-6azacycloalkyl is optionally substituted with one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy.

[0139] The aminopropyl of D is substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy, wherein two C1-4alkyl substituents may combine to form a C2-6cycloalkane, for example, cyclobutane, cyclopentane, cyclohexane, or cycloheptane.

[0140] In some embodiments, D is selected from C3-6azacycloalkyl and aminopropyl, wherein the azacycloalkyl is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy and the aminopropyl is substituted with two C1-4alkyl substituents combined to form a C2-6cycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy.

[0141] In some embodiments, D is aminopropyl, substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy, wherein two C1-4alkyl substituents may combine to form a cyclobutane.

[0142] In some embodiments, D is selected from formulae (Va) to (Vc): wherein * labels the wavy line that dissects the bond to C(O)-NH-B4; m is 1 to 3; o is 0 to 4; and R13and R14are independently H or, as with R15, are as defined above with respect to the optional substituents of D. R13may be selected from H, NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy, such as H. R14may be selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy, such as NH2, hydroxy and C1-4alkyl. R14may be selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy. In specific embodiments, R13is H and R14is selected from NH2, hydroxy, halo, and C1-4alkyl. In specific embodiments, o is 0.

[0143] In some embodiments, D is of formula (Va). In more particular embodiments, D is of formula (Va) and R14is selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy, such as NH2, hydroxy and C1-4alkyl. In yet more particular embodiments, D is of formula (Va) and R14is hydroxy.

[0144] In particular embodiments, D is selected from formula (Vb) or (Vc). In particular embodiments, D is selected from formula (Va) or (Vb), wherein m is 1.

[0145] In some embodiments, R is selected from H and C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl)2, such as H.

[0146] In specific embodiments, the compound of the first aspect is any one selected from formulae (VI 1) to (VI 64), such as (VI 1) to (VI 48):

[0147]

[0148] 

[0149] 

[0150] 

[0151] 

[0152] 

[0153] 

[0154] 

[0155] 

[0156] 

[0157] 

[0158] 

[0159] 

[0160] 

[0161] 

[0162] 

[0163] 

[0164] 

[0165] As described above, the compound of the second aspect may be of formula (1), (2), (3) or (4):

[0166]  wherein A, B1to B7, C1, C2, D and R are as defined in the first aspect; R’ is selected from C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl) and C1-6alkylC(O)NHC1-6alkylNH, R2’ is selected from NHC(O)C1-6alkylNH, C(O)NHC1-6alkylNH, NHC(O)C1-6alkylC(O) and C(O)NHC1-ealkylC(O), and M is a mitochondrial delivery agent or a detectable tag.

[0167] For the avoidance of doubt, the embodiments described in relation to A, B1to B7, C1, C2, D and R of the first aspect may apply mutatis mutandis to the second aspect. For example, A may be of formula (Ila) or (lib), each of B1to B7may be independently selected from formulae (Illa) to (Ilin), each C1and C2may be independently selected from formulae (IVa) to (IVc), D may be selected from formulae (Va) to (Vc), and R may be H.

[0168] In some embodiments, R’ is -C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl)-.

[0169] In some embodiments, R2’ is NHC(O)C1-4alkylNH.

[0170] In some embodiments, the compound of the second aspect is of formula (1), (2) or (3). In some embodiments, the compound of the second aspect is of formula (1) or formula (2).

[0171] Mitochondrial delivery agents are well known in the art. The mitochondrial delivery agent of the second aspect may be any one of those described by Wang et al., in Front. Chem., 2021 , 9, 683220. Due to the presence of proton pumps in the inner mitochondrial membrane, protons in the mitochondrial matrix are pumped out of the mitochondrial matrix, through the inner mitochondrial membrane and into the inter-membrane space, resulting in a net positive charge in the inter-membrane space and a net negative charge in the mitochondrial matrix. Therefore, existing mitochondria-targeting small molecules are typically cationic, cations more favourably entering the mitochondrial matrix owing to the net negative charge within.

[0172] In some embodiments, the mitochondrial delivery agent comprises a moiety selected from a mitochondria-penetrating peptide, triphenylphosphonium, dequalinium, (E)-4- (1 H-lndol-3-ylvinyl)-N-methylpyridineiodide, tetramethylrhodamine ethyl ester, guanidine salt, tetramethylrhodamine methyl ester, rhodamine 19, rhodamine 123 and tetrachlorotetraethyl benzimidazole carbocyanine iodine compounds such as 5, 5', 6,6'- tetrachloro-1 , 1 ',3,3'-tetraethyl-imidacarbocyanine, JC-1 .

[0173] In some examples, the mitochondrial delivery agent is a mitochondria-penetrating peptide. These have alternating cationic and hydrophobic residues, which results in excellent cell permeability and mitochondrial targeting. Many mitochondria-penetrating peptides are known in the art, including cysteine-rich peptides and Szeto-Schiller peptides (see, for example, Kim et al. , Biochemistry, 2020, 59 (3); and Szeto and Schiller, Pharm. Res., 2011 , 28 (11), 2669-2679). The mitochondria-penetrating peptide may comprise cyclohexylalanine and D-arginine amino acid residues. For example, the mitochondria-penetrating peptide may comprise alternating cyclohexylalanine and D- arginine amino acid residues, e.g. two of each residue.

[0174] The structures of specific examples of mitochondria-penetrating peptides are shown below:

[0175] wherein the wavy line of MMP-1 bisects the bond to a carbon atom to which M is attached, and the wavy line of MMP-2 bisects the bond to a nitrogen atom to which M is attached. In particular embodiments, MMP-1 is attached to A and MMP-2 is attached to NH-C(O)-B.

[0176] For example, the compound of the second aspect may be of formula (VII 1):

[0177] Where M is a detectable tag, it may be any tag that is capable of detection on entry of the compound into a cell and / or a mitochondria and that also allows the compound to bind to its target DNA sequence. In some embodiments, where M is a detectable tag, it is selected from a luminescent tag (such as a fluorescent tag, see for example T. Best et al., Proc. Natl. Acad. Sci. U. S. A. 2003, 100, 12063), an affinity tag (see, for example, M. Kimple et al., Curr. Protoc. Protein Sci. 2013;73:9.9.1-9.9.23, and C. Anandhakumar et al. ChemBioChem, 15, 2647-2651 (2014)) and an epitope tag (see, for example, J. Jarvik and C. Telmer. Annu Rev Genet. 1998;32:601-18, and A. Mohammed et al., Am. Chem. Soc. 2023, 145, 45, 24568-24579). In specific embodiments, M is a fluorescent tag, such as a fluorotag (e.g. a fluorescein isothiocyanate), a green fluorescent protein, a red fluorescent protein, HaloTag, SNAP-tag or CLIP-tag.

[0178] The structure of a specific example of a detectable tag is shown below:

[0179]

[0180] For example, the compound of the second aspect may be of formula (VII 2):

[0181] As described above, provided in the third aspect is a mitochondria-targeting capsule comprising a compound of the first and / or second aspect. For the avoidance of doubt, the embodiments described above in relation to A, B1to B7, C1, C2, D and M may apply mutatis mutandis to the third aspect. For example, A may be of formula (Ila) or (lib), each of B1to B7may be independently selected from formulae (Illa) to (Ilin), each C1and C2may be independently selected from formulae (IVa) to (IVc), D may be selected from formulae (Va) to (Vc), and M, when present, may be a fluorescent tag.

[0182] Mitochondria-targeting capsules are well known in the art, and are included in reviews of mitochondria-targeting drug delivery systems such as those by Buchke et al., “Mitochondria-Targeted, Nanoparticle-Based Drug-Delivery Systems: Therapeutics for Mitochondrial Disorders”, Life, 2022, 12, no. 5: 657; and Khan et al., “Mitochondrial Dysfunction: Pathophysiology and Mitochondria-Targeted Drug Delivery Approaches”, Pharmaceutics, 2022, 14(12):2657, https: / / doi.org / 10.3390 / pharmaceutics14122657. The mitochondria-targeting capsule may be a nanosystem, i.e. it may have a diameter of about 50 to about 900 nm, such as about 100 to about 500 nm. The capsule may be selected from a liposome, DQAsome, polymer nanoparticle and inorganic nanoparticle. Typically, the capsule is a liposome, which comprises at least one lipid bilayer. The liposome may be selected from the group consisting of small unilamellar vesicles (SUV), large unilamellar vesicles (LUV), multilamellar vesicle (MLV), and multivesicular vesicles (MW).

[0183] In some embodiments, the mitochondria-targeting capsule is a MITO-porter, such as any one of the MITO-porters described by Yamada et al. in Biochimica et Biophysica Acta (BBA) - Biomembranes, 1778, 2, 2008, 423-432, or Kawamura et al. in Molecular Therapy: Nucleic Acids, 20, 2020, 688-698. Typically, the MITO-porter comprises octaarginine (R8), which is able to target the mitochondrial membrane. In some embodiments, the MITO-porter comprises 1 ,2-dioleoyl-sn-glycero-3-phospho ethanolamine (DOPE), sphingomyelin (SM) and R8.

[0184] MITO-porters comprising one or more of the compounds disclosed herein may be synthesised by slowly titrating with vigorous agitation a solution, such as an ethanol solution, comprising the constituents of the MITO-porter with another solution comprising the one or more compounds to be encapsulated and diluting with further solvent, such as ethanol. The solution may then be concentrated to obtain the one or more compounds disclosed herein encapsulated with the MITO-porter.

[0185] As described above, provided in the fourth aspect is a pharmaceutical composition comprising one or more compounds of the first and / or second aspects of the disclosure and / or a mitochondria-targeting capsule of the third aspect of the disclosure and a pharmaceutically acceptable excipient. For the avoidance of doubt, the embodiments described above in relation to A, B1to B7, C1, C2, D, R, R’ and M and the capsule may apply mutatis mutandis to the fourth aspect. For example, A may be of formula (Ila) or (lib), each of B1to B7may be independently selected from formulae (Illa) to (Ilin), each C1and C2may be independently selected from formulae (IVa) to (IVc), D may be selected from formulae (Va) to (Vc), R (when present) may be H, R’ (when present) may be -C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl)-, M (when present) may be a fluorescent tag, and the capsule, when present, may be a MITO-porter.

[0186] In such pharmaceutical compositions, the one or more compounds of the first and / or second aspects of the disclosure and / or a mitochondria-targeting capsule of the third aspect of the disclosure may be suitably formulated such that it / they can be introduced into the environment of a cell.

[0187] Pharmaceutically acceptable excipients, or carriers, are well known to those skilled in the art and include, but are not limited to, phosphate buffer solutions and / or saline. Pharmaceutically acceptable carriers may be aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's or fixed oils. Preservatives and other additives may also be present, such as, for example, antimicrobials, antioxidants, chelating agents, inert gases and the like.

[0188] In addition to the aforementioned carrier ingredients the pharmaceutical compositions described above may alternatively or additionally include, an appropriate one or more additional excipients, or carrier ingredients, such as diluents, buffers, flavouring agents, binders, surface active agents, thickeners, lubricants, preservatives (including antioxidants) and the like, and substances included for the purpose of rendering the formulation isotonic with the blood of the intended recipient.

[0189] Pharmaceutical compositions may be present in any formulation typical for the administration of a pharmaceutical compound to a subject. Representative examples of typical formulations include, but are not limited to, capsules, granules, tablets, powders, lozenges, suppositories, pessaries, nasal sprays, gels, creams, ointments, sterile aqueous preparations, sterile solutions, aerosols, implants etc.

[0190] Pharmaceutical compositions are formulated to be compatible with the intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral, transdermal, topical, transmucosal, vaginal and rectal administration.

[0191] The pharmaceutical compositions may include those suitable for oral, parenteral (including subcutaneous, intradermal, intramuscular and intravenous), topical (including dermal, buccal and sublingual), rectal, nasal and pulmonary administration e.g., by inhalation. The composition may, where appropriate, be conveniently presented in discrete dosage units and may be prepared by any of the methods well known in the art of pharmacy. Methods typically include the step of bringing into association an active with liquid carriers or finely divided solid carriers or both and then, if necessary, shaping the product into the desired formulation.

[0192] Pharmaceutical compositions suitable for oral administration wherein the carrier is a solid are most preferably presented as unit dose formulations such as boluses, capsules or tablets each containing a predetermined amount of active. A tablet may be made by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine an active compound in a free-flowing form such as a powder or granules optionally mixed with a binder, lubricant, inert diluent, lubricating agent, surface-active agent or dispersing agent. Moulded tablets may be made by moulding an active compound with an inert liquid diluent. Tablets may be optionally coated and, if uncoated, may optionally be scored. Capsules may be prepared by inserting an active, either alone or in admixture with one or more accessory ingredients, into the capsule shells and then sealing them in the usual manner. Cachets are analogous to capsules wherein an active together with any accessory ingredient(s) is sealed in a rice paper envelope. The active may also be formulated as dispersible granules, which may for example be suspended in water before administration, or sprinkled on food. The granules may be packaged, e.g., in a sachet. Compositions suitable for oral administration wherein the carrier is a liquid may be presented as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water liquid emulsion. Compositions for oral administration include controlled release dosage forms, e.g., tablets wherein an active compound or capsule is formulated in an appropriate release-controlling matrix, or is coated with a suitable release-controlling film.

[0193] Pharmaceutical compositions suitable for parenteral administration include sterile solutions or suspensions of an active in aqueous or oleaginous vehicles. Injectable preparations may be adapted for bolus injection or continuous infusion. Such preparations are conveniently presented in unit dose or multi-dose containers, which are sealed after introduction of the formulation until required for use. Alternatively, the compound or capsule may be in powder form, which is constituted with a suitable vehicle, such as sterile, pyrogen-free water, before use. The pharmaceutical composition may also be formulated as a long-acting depot preparation, which may be administered by intramuscular injection or by implantation, e.g., subcutaneously or intramuscularly. Depot preparations include, for example, suitable polymeric or hydrophobic materials, or ion-exchange resins.

[0194] Pharmaceutical compositions suitable for topical formulation may be provided for example as gels, creams or ointments.

[0195] The compounds or capsules described herein may be present in the pharmaceutical compositions as a pharmaceutically and / or physiologically acceptable salt, solvate or derivative.

[0196] As used herein, the term "pharmaceutically acceptable salt" refers to those salts, which are generally considered suitable for use in medicine (including in a veterinary context). For example, pharmaceutically acceptable salts may be those which can be contacted with the tissues of a mammalian subject (e.g. humans) without undue toxicity, irritation, allergic response or the like. By way of further example of suitable pharmaceutically acceptable salts, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, the entire contents of which are incorporated herein by reference.

[0197] Representative examples of pharmaceutically and / or physiologically acceptable salts of the compounds or capsules of the disclosure may include, but are not limited to, acid addition salts formed with organic carboxylic acids such as acetic, lactic, tartaric, maleic, citric, pyruvic, oxalic, malonic, fumaric, oxaloacetic, isethionic, lactobionic and succinic acids; organic sulfonic acids such as methanesulfonic, ethanesulfonic, benzenesulfonic and p-toluenesulfonic acids and inorganic acids such as hydrochloric, hydrobromic, sulfuric, perchloric, phosphoric and sulfamic acids. Other pharmaceutically acceptable salts include (but are not limited to) adipate, alginate, ascorbate, aspartate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2- hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3- phenylpropionate, pivalate, propionate, stearate, thiocyanate, undecanoate, valerate salts, and the like.

[0198] In some examples, salts that may be derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(Ci-4alkyl)4 salts. Representative alkali or alkaline earth metal salts include, but are not limited to, sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts may include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.

[0199] Pharmaceutically and / or physiologically functional derivatives of compounds or capsules of the present disclosure are derivatives, which may be converted in the body into the parent compound or capsule. Such pharmaceutically and / or physiologically functional derivatives may also be referred to as "pro-drugs" or "bioprecursors". Pharmaceutically and / or physiologically functional derivatives of compounds or capsules of the present disclosure may include hydrolysable esters or amides, particularly esters, in vivo.

[0200] It may be convenient or desirable to prepare, purify, and / or handle a corresponding pharmaceutically and / or physiologically acceptable solvate of the compound or capsule described herein, which may be used in the any one of the uses / methods described. The term solvate is used herein to refer to a complex of solute, such as a compound or salt of the compound, and a solvent. If the solvent is water, the solvate may be termed a hydrate, for example a mono-hydrate, di-hydrate, tri-hydrate etc, depending on the number of water molecules present per molecule of substrate.

[0201] As described above, provided in the fifth aspect is a compound of the first or second aspect of the disclosure, a mitochondria-targeting capsule of the third aspect of the disclosure, or pharmaceutical composition of the fourth aspect of the disclosure for use in a method of treatment. Provided in the sixth aspect is a compound of the first or second aspect of the disclosure, a mitochondria-targeting capsule of the third aspect of the disclosure, or pharmaceutical composition of the fourth aspect of the disclosure for use in a method of treating or preventing mitochondrial dysfunction or a disease or condition associated therewith. Provided in the seventh aspect is a method of treating or preventing mitochondrial dysfunction or a disease or condition associated therewith in a subject comprising administering a therapeutically effective amount of a compound of the first or second aspect, a mitochondria-targeting capsule of the third aspect, or a pharmaceutical composition of the fourth aspect to the subject. Provided in the eighth aspect is the compound of the first or second aspect, the mitochondria-targeting capsule of the third aspect, or pharmaceutical composition of the fourth aspect for use in medicine or as a medicament.

[0202] For the avoidance of doubt, the embodiments described above in relation to A, B1to B7, C1, C2, D, R, R’, M, the capsule and the pharmaceutical excipient may apply mutatis mutandis to the fifth, sixth, seventh and / or eighth aspects. For example, A may be of formula (Ila) or (lib), each of B1to B7may be independently selected from formulae (Illa) to (Ilin), each C1and C2may be independently selected from formulae (IVa) to (IVc), D may be selected from formulae (Va) to (Vc), R (when present) may be H, R’ (when present) may be -C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl)-, M (when present) may be a fluorescent tag, the capsule, when present, may be a MITO-porter, and the pharmaceutical excipient, when present, may be a pharmaceutically acceptable salt.

[0203] As described above, mitochondria supply 90% of the energy needed by a cell by generating adenosine triphosphate through oxidative phosphorylation of adenosine diphosphate. The disease or condition associated with mitochondrial dysfunction may therefore be any symptom, e.g. human symptom, arising from sub-normal adenosine triphosphate (ATP) levels. For example, it may be any symptom arising from premature and progressively declining ATP levels (see, for example, Johnson, Jinnah and Kamatani, “Shortage of Cellular ATP as a Cause of Diseases and Strategies to Enhance ATP”, Front. Pharmacol., 2019, 10, https: / / www.frontiersin.Org / articles / 10.3389 / fphar.2019.00098).

[0204] The disease or condition associated with mitochondrial dysfunction may be any of those described by Chinnery in “Primary Mitochondrial Disorders Overview”, 2000, in: Adam MP, Feldman J, Mirzaa GM, et al., editors. GeneReviews® [Internet], Seattle (WA): University of Washington, Seattle; 1993-2023, or by Pieczenik and Neustadt in Experimental and Molecular Pathology, 83, 1 , 2007, 84-92.

[0205] The disease or condition associated with mitochondrial dysfunction may be mitochondrial myopathy, mitochondrial diabetes, schizophrenia, Leber’s hereditary optic neuropathy (LHON), Leigh syndrome, neuropathy, ataxia, retinitis pigmentosa, and ptosis (NARP), or myoneurogenic gastrointestinal encephalopathy (MNGIE). Alternatively, or in addition, treatment or prevention may be of any form of cardiovascular disease, cancer, obesity and / or insulin resistance (e.g. type 2 diabetes) that has been caused by a mitochondrial dysfunction.

[0206] In some embodiments, the mitochondrial dysfunction to be treated is related to oxidative stress.

[0207] In some embodiments, the disease or condition associated with mitochondrial dysfunction is a neurodegenerative disease. The neurodegenerative disease may be dementia, Alzheimer’s disease (AD), Huntington’s disease (HD), amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), Parkinson’s disease (PD), multiple system atrophy (MSA), Batten disease, or Creutzfeldt-Jakob disease (CJD).

[0208] Provided in the ninth embodiment is a compound selected from formulae (i), (ii) and (iii): wherein:

[0209] P is an amine protecting group;

[0210] LG is a leaving group; r1is selected from C1-4alkylC(O)OC1-4alkyl, C1-4alkylC1-4alkoxy, C1-4alkylC1-4alkoxyC1-4alkoxy, C1-4alkylcyano and C1-4alkylO-P1, wherein P1is an alcohol protecting group; r2is selected from C1-4alkyl and C1-4haloalkyl; r3is selected from C1-4alkyl, halo and C1-4haloalkyl; r4is selected from C1-4haloalkyl, C1-4alkylO-P1and formula (iiia):

[0211] P (iiia); s1and s2are each independently selected from C1-4alkyl and C1-4haloalkyl; and ns1 and ns2 are each independently selected from 0 to 2.

[0212] As described above, particular isolatable intermediates such as those within the scope of formulae (i), (ii) and (iii), used to synthesise the compounds of the first or second aspects of the invention have been found by the inventors to be particularly useful as components of the compounds of the first or second aspects of the invention. For example, particular intermediates, when included in the compounds of the first or second aspects, have been found to give rise to favourable properties, such as increased solubility and / or selectivity for binding to a particular sequence of mtDNA.

[0213] P is an amine protecting group. Such groups are well known in the art and bind to an amine group to reduce the reactivity of the amine so that reaction of the amine is prevented in subsequent chemical processes. After the subsequent chemical processes have been carried out, the amine protecting group can be easily removed under suitable conditions, which are well known in the art. For a review of amine protecting groups, see A. Isidro-Llobet et al., Chem. Rev. 2009, 109, 2455-2504. In some embodiments, P is a species capable of forming a substituted amide, including a carbamate, a substituted sulfonamide, or a substituted secondary amine on reaction with an amino (-NH2) group.

[0214] In some embodiments, P is selected from -CC^r5, -COr5, -SC^r6, and -C(r7)s, wherein: r5is selected from C1-4alkyl, C1-4alkylfluorenyl, C1-4alkenyl and C1-4alkylphenyl; r6is selected from phenyl, C1-4alkyl and C1-4haloalkyl, wherein the phenyl is optionally substituted with one or more substituents selected from C1-4alkyl, nitro, halo and C1-4haloalkyl; and each r7is independently selected from phenyl, H and C1-4alkyl, wherein the phenyl is optionally substituted with one or more substituents selected from C1-4alkyl, halo and C1-4haloalkyl. Typically, at least one r7is an optionally substituted phenyl.

[0215] In some embodiments, r6is phenyl substituted with one or more substituents selected from C1-4alkyl and nitro. In some embodiments, at least one r7is phenyl and the other two are independently selected from phenyl and H. In some embodiments, P is a species capable of forming a carbamate on reaction with an amino (-NH2) group. In some embodiments, P is -CC^r5, wherein r5is selected from C1-4alkyl, C1-4alkylfluorenyl, and C1-4alkenyl.

[0216] P1is an alcohol protecting group. Such groups are also well known in the art and bind to an alcohol group to an alcohol group to reduce the reactivity of the alcohol so that reaction of the alcohol is prevented in subsequent chemical processes. After the subsequent chemical processes have been carried out, the alcohol protecting group can be easily removed under suitable conditions, which are well known in the art. For a review of alcohol protecting groups, see M. Lalonde and T. H. Chan, Synthesis 1985; 1985(9): 817-845. In some embodiments, P1is a species capable of forming a silyl ether on reaction with an alcohol group. In some embodiments, P1is a trisubstituted silyl such as trimethylsilyl, triethylsilyl, triisopropylsilyl, tert-butyldimethylsilyl, di-tert- butylmethylsilyl or tert-butyldiphenylsilyl.

[0217] LG is a leaving group. This is defined by the IUPAC as an atom or group of atoms that detaches from the main or residual part of a substrate during a reaction or elementary step of a reaction. Herein, the leaving group is capable of detaching from a carbonyl. Such groups are well known to those skilled in the art and include halo, haloalkyl, alkoxy, haloalkoxy and carboxylate groups. In some embodiments, the leaving group is selected from hydroxy, C1-4haloalkyl (such as trihalomethyl, e.g. trichloromethyl), C1-4alkoxy, C1-4haloalkoxy and halo (such as chloro).

[0218] In some embodiments, wherein ns1 and ns2 are each 0.

[0219] In some embodiments, r1is selected from C2-4alkylC(O)OC1-2alkyl, C2-4alkylC1-2alkoxy, C2-6alkylC2-6alkoxyC2-6alkoxy, C2-6alkylcyano and C2-4alkylO-P1, wherein P1is a trisubstituted silyl such as trimethylsilyl, triethylsilyl, triisopropylsilyl, tert- butyldimethylsilyl, di-tert-butylmethylsilyl or tert-butyldiphenylsilyl.

[0220] In some embodiments, r2is selected from C1-2alkyl and C1-2haloalkyl, such as methyl.

[0221] In some embodiments, r3is selected from C2-6alkyl, halo and C2-6haloalkyl, such as methyl and bromo. In some embodiments, r4is selected from C2-6haloalkyl, C2-4alkylO-P1and formula (iiia), wherein wherein P1is a trisubstituted silyl such as trimethylsilyl, triethylsilyl, triisopropylsilyl, tert-butyldimethylsilyl, di-tert-butylmethylsilyl or tert-butyldiphenylsilyl and P is -CO2C1-6alkyl.

[0222] In particular embodiments, the compound is selected from formulae (iva) to (ivk):

[0223]

[0224] Provided in the tenth aspect is use of a compound of the ninth aspect in the synthesis of a polyamide, such as a polyamide of the first or second aspect.

[0225] For the avoidance of doubt, the embodiments described above in relation to P, LG, r1, r2, r3, r4, s1, s2, ns1 and ns2 may apply mutatis mutandis to the tenth aspect.

[0226] It will be appreciated that the compounds of the present disclosure may exist in different stereoisomeric forms. The present disclosure includes within its scope the use of all stereoisomeric forms, or the use of a mixture of stereoisomers of the bifunctional molecules. By way of example, where the bifunctional molecule comprises one or more chiral centres, the present disclosure encompasses each individual enantiomer of the bifunctional molecule as well as mixtures of enantiomers including racemic mixtures of such enantiomers. By way of further example, where the bifunctional molecule comprises two or more chiral centres, the present disclosure encompasses each individual diastereomer of the bifunctional molecule, as well as mixtures of the various diastereomers. Unless otherwise indicated, the various structures shown herein encompass all isomeric (e.g. enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure). For example, the present disclosure embraces the R and S configurations for each asymmetric centre, and Z and E double bond isomers. Therefore, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are to be understood to be within the scope of the present disclosure. Additionally, unless otherwise stated, where present, all tautomeric forms of the bifunctional molecules described herein are to be understood to be within the scope of the present disclosure.

[0227] Additionally, unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, bifunctional molecules as described herein in which one or more hydrogen atoms have been replaced by deuterium or tritium, or in which one or more carbon atoms have been replaced by a13C- or14C-enriched carbon are to be understood to within the scope of the present disclosure. Such molecules may be useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents in accordance with the present disclosure. By way of further example, a bifunctional molecule as described herein, may be substituted with one or more deuterium atoms.

[0228] Each and every patent and non-patent reference referred to herein is hereby incorporated by reference in its entirety, as if the entire contents of each reference were set forth herein in their entirety.

[0229] Aspects and embodiments of the invention are further described in the following clauses:

[0230] 1. A compound of formula (I): wherein:

[0231] A is a 5-membered heterocycle optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl; each B1to B7is independently a 5- or 6-membered heterocycle, pentanone, benzene, 5- or 6-membered aliphatic carbocycle or ethan-1 ,2-di-yl, wherein: the 5- or 6- membered heterocycle, pentanone, benzene and 5- or 6-membered aliphatic carbocycle are optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-ehaloalkyl, halo, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl-(5- membered heterocyclyl), C1-4alkyl(C1-4alkoxy)pC1-4alkoxy, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), carboxyl, CO2(C1-4alkyl), CO(C1-4alkyl), C1-4alkylCO2(C1-4alkyl), oxo, C2-6alkynyl, C2-6haloalkynyl, amino, hydroxy, C1-6alkyldiC1-4alkylamino, C1-4alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC3-6azacycloalkyl, C1-4alkylmorpholinyl, C1-4alkylCs- ehaloazacycloalkyl, phenyl, 5-membered heterocyclyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2; and the ethan-1 , 2-di-yl is optionally substituted with any one or more selected from hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy, wherein the C1-4alkyl substituent may combine with the -NH- group adjacent to the ethan-1 , 2-di-yl to form a C3-5azacycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; each C1and C2is independently selected from a 5- or 6-membered N-heterocycle and ethan-1 , 2-di-yl, wherein the 5- or 6-membered N-heterocycle is optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl-(5-membered heterocyclyl), C1-4alkyl(C1-4alkoxy)pC1-4alkoxy, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), carboxyl, CO2(C1-4alkyl), oxo, C2-6alkynyl, C2-6haloalkynyl, amino, hydroxy, C1-4alkyldiC1-4alkylamino, C1-6alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC1-4alkoxyC1-4alkoxy, C1-4alkylCs-6azacycloalkyl, C1-4alkylmorpholinyl, C1-4alkylC3-6haloazacycloalkyl, phenyl, 5- membered heterocyclyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2; and the ethan-1 ,2-di- yl is optionally substituted with any one or more selected from hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy; p is 0 to 5;

[0232] D is selected from C3-6azacycloalkyl and aminopropyl, wherein the azacycloalkyl is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy and the aminopropyl is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy, wherein two C1-4alkyl substituents may combine to form a C2-6cycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; and

[0233] R is selected from H, C1-6alkylC(O)NHCi.6alkylN(Ci.ealkyl)2, C1-4alkylC(O)NHC1-6alkylN(C1-6alkyl)H and C1-4alkylC(O)NHC1-4alkylNH2.

[0234] 2. The compound of clause 1 , wherein A is an optionally substituted N-heteroaryl.

[0235] 3. The compound of clause 2, wherein the N-heteroaryl comprises 1 or 2 nitrogen atoms.

[0236] 4. The compound of any one of clauses 1 to 3, wherein A is an optionally substituted pyrrole or imidazole.

[0237] 5. The compound of any one of clauses 1 to 4, wherein A is of formula (Ila) or (lib): wherein:

[0238] R1and R2are each independently selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl; and

[0239] R3and R4are each independently selected from hydrogen, halo, C1-4alkyl and C1-ehaloalkyl.

[0240] 6. The compound of clause 5, wherein:

[0241] R1is selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-2alkylphenyl, C1-4alkylCs- ecycloalkyl, C2-6azacycloalkyl, thiophenyl and thiazolyl;

[0242] R2is C1-4alkyl; and

[0243] R3and R4are each independently selected from H and halo.

[0244] 7. The compound of clause 5, wherein A is of formula (Ila) and R1is selected from C2- ealkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl. 8. The compound of clause 5, wherein A is of formula (Ila) and R1is C1-4alkyl.

[0245] 9. The compound of any one preceding clause, wherein each of B1to B7is independently an optionally substituted 5- or 6-membered heterocycle comprising one or more heteroatoms selected from nitrogen, sulphur and oxygen; an optionally substituted pentanone; an optionally substituted benzene; an optionally substituted 5-membered aliphatic carbocycle; or an optionally substituted ethan-1 ,2-di-yl.

[0246] 10. The compound of any one preceding clause, wherein each of B1to B7is independently selected from an optionally substituted pyrrole, pyrazole, furazan, thiophene, pyrrolidine, tetra hydrofuran, imidazolidine, piperidine, and oxazolidine; an optionally substituted pentanone; an optionally substituted benzene; an optionally substituted cyclopentane; and an optionally substituted ethan-1 ,2-di-yl. 11. The compound of any one preceding clause, wherein each of B1to B7is independently selected from formulae (Illa) to (Ilin):

[0247] wherein:

[0248] * labels the wavy line that bissects the bond to carbonyl;

[0249] R5and R7are each independently selected from H, C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, Ci.6alkylC1-4alkoxy, C1-ealkylcarboxyl, C1-4alkylCO2(C1-4alkyl), C1-4alkyldiC1-4alkylamino, C1-4alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC1-4alkoxyC1-4alkoxy, C1-4alkylC3-6azacycloalkyl, C1-4alkylC1-sheteroaryl, and C1-4alkylC3-6haloazacycloalkyl;

[0250] R6is selected from C1-4alkyl, C1-4haloalkyl and halo;

[0251] R8is selected from H, C1-4alkyl, C1-4haloalkyl, halo, C2-ealkynyl, C2.ehaloalkynyl, amino and phenyl; each X is independently selected from NR9, O and C(O); each R9is independently selected from H, C1-4alkyl, C(O)C1-4alkyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2; each R10is independently selected from C1-4alkyl, C1-4haloalkyl, halo, hydroxy, amino and C1-4alkoxy; each R11and R12is independently selected from H, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, NH2, phenyl and C1-4alkoxy, wherein the C1-4alkyl substituent may combine with the -NH- group adjacent to the ethan-1 ,2-di-yl to form a C3-5azacycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; n is 0 to 2; n1 is 0 or 1 ; and n2 is 0 to 3.

[0252] 12. The compound of clause 11 , wherein:

[0253] R5is selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl(C1-4alkoxy)i-5, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), C1-4alkyldiC1-4alkylamino, C1-4alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC3-6azacycloalkyl, C1-4alkylC1-3heteroaryl, and C1-4alkylC3-6haloazacycloalkyl;

[0254] R6and R7are each independently C1-4alkyl;

[0255] R8is selected from H, C1-4alkyl, C1-4haloalkyl, halo, C2-6alkynyl, C2-6haloalkynyl, amino and phenyl;

[0256] R11is H or C1-4alkyl, wherein the C1-4alkyl substituent may combine with the -NH- group adjacent to the ethan-1 ,2-di-yl to form a C3-5azacycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; and

[0257] R12is selected from H, hydroxy, C1-4alkyl, NH2, and phenyl.

[0258] 13. The compound of clause 11 or clause 12, wherein:

[0259] R5is selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkyl(C1-4alkoxy)i-5, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), and C1-4alkylcyano; n is 0;

[0260] R7is C1-4alkyl; and

[0261] R8is H.

[0262] 14. The compound of any one of clauses 11 to 13, wherein B1, B3, B4and B7are of formula (Illa).

[0263] 15. The compound of clause 14, wherein R5of B4and B7are each independently selected from C1-4alkyl, such as methyl.

[0264] 16. The compound of any one preceding clause, wherein B3is of formula (Illa) and R5of B3is selected from C1-4alkylol, C1-4alkyl(C1-4alkoxy)i-5, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), and C1-4alkylcyano, such as C1-4alkylCO2(C1-4alkyl).

[0265] 17. The compound of clause 14 or clause 15, wherein R5of B3is selected from C1-4alkyl, C1-4alkylCO2(C1-4alkyl), and C1-4alkylcarboxyl.

[0266] 18. The compound of any one preceding clause, wherein B2is selected from: formula (Illa), wherein R5is selected from C1-4alkylol, C1-4alkyl(C1-4alkoxy)i-5, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), and C1-4alkylcyano; formula (lllc), wherein R7is C1-4alkyl and R8is selected from C1-4alkyl, C1-

[0267] 4haloalkyl and halo; formula (Hid); formula (Illg), wherein X is NR9, wherein R9is selected from H and CO(C1-4alkyl) and each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo; formula (IIIm), wherein one of R11and R12is ethyl and combines with the -NH- group adjacent to the ethan-1 ,2-di-yl to form pyrrolidinyl that is optionally substituted with any one or more selected from C1-4alkyl, halo and C1-4haloalkyl; and formula (Ilin), wherein each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo.

[0268] 19. The compound of any one of clauses 11 to 18, wherein B5is of formula (Illa), (lllc), (Illg) or (IIIm).

[0269] 20. The compound of clause 19, wherein:

[0270] R5of B5is selected from C1-4alkyl, C1-4alkylcarboxyl and C1-4alkylCs-6azacycloalkyl;

[0271] R7and R8of B5are each independently selected from H and C1-4alkyl;

[0272] X is NH and each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo and

[0273] R11and R12of B5are each independently selected from H and C1-4alkyl.

[0274] 21 . The compound of any one preceding clause, wherein B5is: of formula (Illg), wherein X is NH and each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo; or of formula (IIIm), wherein R11is C1-4alkyl and R12is H or C1-4alkyl.

[0275] 22. The compound of any one of clauses 11 to 21 , wherein B6is selected from formula (Illa), (Illg) and (IIIm).

[0276] 23. The compound of clause 22, wherein:

[0277] R5of B6is C1-4alkyl, such as methyl, or C1-4alkyl(C1-4alkoxy)i-5;

[0278] R11of B6is H; and

[0279] R12of B6is selected from H, hydroxy, C1-4alkyl, NH2, and phenyl. 24. The compound of any one preceding clause, wherein the 5- or 6-membered N- heterocycle of C1and C2is pyrrolidine.

[0280] 25. The compound of any one preceding clause, wherein each C1and C2is independently selected from formulae (IVa) to (IVc): wherein:

[0281] * labels the wavy line that dissects the bond to carbonyl;

[0282] R5is C1-4alkyl, such as methyl; and each R11and R12is independently selected from H, hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy.

[0283] 26. The compound of clause 25, wherein R11of formula (IVc) is H.

[0284] 27. The compound of any one of clauses 1 to 25, wherein C1is of formula (IVc), R11is H and R12is independently selected from C1-4alkyl, C1-4haloalkyl and C1-4alkoxy.

[0285] 28. The compound of clause 25 or clause 26, wherein C1and C2are independently selected from formulae (IVa) and (IVc).

[0286] 29. The compound of any one preceding clause, wherein D is selected from C3-6azacycloalkyl and aminopropyl, wherein the azacycloalkyl is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy and the aminopropyl is substituted with two C1-4alkyl substituents combined to form a C4-ycycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy.

[0287] 30. The compound of any one of clauses 1 to 28, wherein D is selected from formulae (Va) to (Vc):

[0288] wherein:

[0289] R13is selected from H, NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy;

[0290] R14is selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy;

[0291] R15is selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; m is 1 to 3; and o is 0 to 4.

[0292] 31 . The compound of clause 30, wherein:

[0293] R13is H;

[0294] R14is selected from NH2, hydroxy, halo, and C1-4alkyl; m is 1 ; and o is 0.

[0295] 32. The compound of clause 30 or clause 31 , wherein D is of formula (Va).

[0296] 33. The compound of any one preceding clause, wherein R is selected from H, and C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl)2, such as H.

[0297] 34. A compound of formula (1), (2), (3), or (4):

[0298]  wherein:

[0299] A, B1to B7, C1, C2, D and R are as defined in any one preceding clause;

[0300] R’ is selected from C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl) and C1-6alkylC(O)NHCi.6alkylNH;

[0301] R2’ is selected from NHC(O)C1-6alkylNH, C(O)NHC1-6alkylNH, NHC(O)C1-6alkylC(O) and C(O)NHC1-6alkylC(O); and

[0302] M is a mitochondrial delivery agent or a detectable tag.

[0303] 35. A mitochondria-targeting capsule comprising a compound of any one preceding clause.

[0304] 36. The mitochondria-targeting capsule of clause 35, which is a MITO-porter comprising octaarginine.

[0305] 37. A pharmaceutical composition comprising a compound of any one of clauses 1 to 34 or a mitochondria-targeting capsule of clause 35 or clause 36 and a pharmaceutically acceptable excipient.

[0306] 38. The compound of any one of clauses 1 to 34, mitochondria-targeting capsule of clause 35 or clause 36, or pharmaceutical composition of clause 37 for use in a method of treatment.

[0307] 39. The compound of any one of clauses 1 to 34, mitochondria-targeting capsule of clause 35 or clause 36, or pharmaceutical composition of clause 37 for use in a method of treating or preventing mitochondrial dysfunction or a disease or condition associated therewith.

[0308] 40. A method of treating or preventing mitochondrial dysfunction or a disease or condition associated therewith in a subject comprising administering a therapeutically effective amount of a compound of any one of clauses 1 to 34, mitochondria-targeting capsule of clause 35 or clause 36, or pharmaceutical composition of clause 37 to the subject.

[0309] 41 . The compound of any one of clauses 1 to 34, mitochondria-targeting capsule of clause 35 or clause 36, or pharmaceutical composition of clause 37 for use in medicine or as a medicament.

[0310] 42. A compound selected from formulae (i), (ii) and (iii): wherein:

[0311] P is an amine protecting group;

[0312] LG is a leaving group; r1is selected from C1-4alkylC(O)OC1-4alkyl, C1-4alkylC1-4alkoxy, C1-4alkylC1-

[0313] 4alkoxyC1-4alkoxy, C1-4alkylcyano and C1-4alkylO-P1, wherein P1is an alcohol protecting group; r2is selected from C1-4alkyl and C1-4haloalkyl; r3is selected from C1-4alkyl, halo and C1-4haloalkyl; r4is selected from C1-4haloalkyl, C1-4alkylO-P1and formula (iiia):

[0314] P (iiia); s1and s2are each independently selected from C1-4alkyl and C1-4haloalkyl; and ns1 and ns2 are each independently selected from 0 to 2.

[0315] 43. The compound of clause 42, wherein P is selected from -CC^r5, -COr5, -SC^r6, and -C(r7)s, wherein: r5is selected from C1-4alkyl, C1-4alkylfluorenyl, C1-4alkenyl and C1-4alkylphenyl; r6is selected from phenyl, C1-4alkyl and C1-4haloalkyl, wherein the phenyl is optionally substituted with one or more substituents selected from C1-4alkyl, nitro, halo and C1-4haloalkyl; and each r7is independently selected from phenyl, H and C1-4alkyl, wherein the phenyl is optionally substituted with one or more substituents selected from C1-4alkyl, halo and C1-4haloalkyl. Typically, at least one r7is an optionally substituted phenyl.

[0316] 44. The compound of clause 42 or clause 43, wherein P1is a trisubstituted silyl such as trimethylsilyl, triethylsilyl, triisopropylsilyl, tert-butyldimethylsilyl, di-tert- butylmethylsilyl or tert-butyldiphenylsilyl.

[0317] 45. The compound of any one of clauses 42 to 44, wherein LG is selected from hydroxy, C1-4haloalkyl, C1-4alkoxy, C1-4haloalkoxy and halo.

[0318] 46. The compound of any one of clauses 42 to 45, wherein ns1 and ns2 are each 0.

[0319] 47. Use of a compound of any one of clauses 42 to 45 in the synthesis of a polyamide.

[0320] 47. The compound of any one of clauses 42 to 46, wherein the compound is selected from formulae (iva) to (ivk):

[0321]  BRIEF DESCRIPTION OF FIGURES

[0322] Fig. 1 contains bar charts showing the full screening results of high throughput screening of polyamides for transcriptional inhibition.

[0323] Fig. 2 contains bar charts showing representative polyamide results of transcriptional inhibition to demonstrate the effect of different building blocks.

[0324] EXAMPLES

[0325] The present invention will now be described in detail with reference to the following nonlimiting examples.

[0326] SYNTHESIS

[0327] Synthesis of monomers

[0328] General procedures

[0329] All commercially available reagents and solvents used were obtained from Sigma- Aldrich, Fluorochem Fisher Scientific, Acros, Alfa Aesar, Apollo scientific and Advanced Chem Blocks and used without further purification. Air- or moisture-sensitive reactions were carried out under argon or nitrogen atmosphere. Microwave reactions were carried out using a Biotage Initiator system. Flash chromatography was performed using a Biotage SP4 automated chromatography system using silica stationary phase (Fisher Scientific, 60 A, 35-70 micron; detection wavelength: 254 nm; monitoring: 280 nm) and the mobile phase used are detailed in the text. Reverse phase HPLC purifications were conducted on Shimadzu Prominance HPLC using a semi-preparative (50 x 21.2 mm) Luna 5pm C18 column at 40 °C; flow rate: 6 ml / min; detection wavelength: 254 nm eluting with an acetonitrile / water gradient with 0.1% TFA. NMR spectra were recorded on either a Bruker Avance3 / DPX400 (400 MHz), Bruker DRX500 (500 MHz), Bruker AV400 (400 MHz), Bruker AV500HD (500 MHz) or Bruker AV600 (600 MHz) instrument and analysed using Advanced Chemistry Development Labs (ACD / labs) NMR 87 O B O Cl F N processor 12.00 or MestReNova 10.0 software. Chemical shifts (5) are recorded in parts per million (ppm) relative to an internal solvent reference (tetramethylsilane) and coupling constants (J) in Hertz (Hz). Splitting patterns were indicated as singlet (s), broad singlet (br. s), doublet (d), doublet of doublet (dd), triplet (t), quartet (q) and multiplet (m). LCMS was carried out on an Agilent Technologies 1220 series LC system with Agilent 6100 series quadrupole mass spectrometer in ESI / APCI mode. Separation was achieved with an Agilent Eclipse C18 4.6x50 mm column; flow rate:1 ml / min; detection:254 nm; sample volume: 10 pl; mobile phase: acetonitrile / 5mM ammonium acetate :water / 5mM ammonium acetate; 5%, 1.48 min; 5-100%, 8 min; 100%, 13.5 min; 100-5%, 16.5 min; 18 min. HRMS was carried out on an Exactive (Thermo scientific) or LTQ orbitrap (Thermo scientific).

[0330] Thin-layer chromatography (TLC) was carried out on aluminium-backed SiC>2 plates (Merck, silica gel 60, F254) and spots visualised using ultra-violet light (254 nm) or by staining with potassium permanganate. All tested compounds were determined to be >95 % purity by LC-MS and analytical HPLC unless otherwise stated.

[0331] The FMOC-protected cyclopentane monomers used in the synthesis of VI 49, VI 50, VI 57 and VI 58 are commercially available.

[0332] Route to 4-(((Allyloxy)carbonyl)amino)-1-(3-methoxy-3-oxopropyl)-1H-pyrrole-2- carboxylic acid tert-Butyl 1-(3-methoxy-3-oxopropyl)-4-nitro-1H-pyrrole-2-carboxylate

[0333] To a mixture of terf-butyl 4-nitro-1 / 7-pyrrole-2-carboxylate (1.0 g, 4.71 mmol, 1.0 eq), methyl 3-bromopropionate (0.866 g, 0.566 mL, 5.18 mmol, 1.1 eq), potassium carbonate (0.977 g, 7.07 mmol, 1.5 eq) and tetrabutylammonium iodide (0.174 g, 0.471 mmol, 0.1 eq) in a sealed flask flushed with argon was added dimethylformamide (DMF) (5 mL) and the resultant mixture stirred at 80 °C overnight. The reaction was then cooled and quenched by the addition of water and extracted with ethyl acetate (3 X 50 mL). The combined organic layers were washed with brine (3 X 20 mL) and absorbed onto celite. Purification by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 40% percent) afforded the target compound as a yellow oil which solidified on standing (1.27 g, 91%).

[0334] 1H NMR (400 MHz, DMSO) σ 8.22 (d, J = 2.1 Hz, 1 H), 7.25 (d, J = 2.1 Hz, 1 H), 4.57 (t, J = 7.0 Hz, 2H), 3.61 (s, 3H), 2.88 (t, J = 7.1 Hz, 2H), 1.53 (s, 9H)13C NMR (101 MHz, DMSO) σ 170.7, 158.6, 134.2, 128.8, 123.4, 111.9, 81.9, 51.6, 45.1 , 34.6, 27.7. tert-Butyl 4-amino-1-(3-methoxy-3-oxopropyl)-1H-pyrrole-2-carboxylate

[0335] To a mixture of terf-butyl 1-(3-methoxy-3-oxopropyl)-4-nitro-1 / 7-pyrrole-2-carboxylate (1.19 g, 3.99 mmol, 1 eq) and palladium on carbon 10% wt (0.250 g) in a 250 mL round bottom flask, sealed and flushed with argon was added methanol (50 mL) followed by a balloon of hydrogen and the resulting reaction mixture stirred at 45 °covernight. On completion (monitored by LC-MS), the crude reaction mixture was absorbed onto celite under reduced pressure and purified by flash chromatography eluting with a gradient of petroleum ether to 10% methanol in ethyl acetate (0 to 80%) to afford the target compound as a deep red oil (0.98 g, 92%).

[0336] 1H NMR (400 MHz, DMSO) σ 6.37 (d, J = 2.3 Hz, 1 H), 6.19 (d, J = 2.2 Hz, 1 H), 4.12 (t, J = 6.8 Hz, 2H), 3.87 (s, 2H), 3.58 (s, 3H), 2.21 (t, J = 7.5 Hz, 2H), 1.87 (p, J = 7.2 Hz, 2H), 1.47 (s, 9H).

[0337] 13C NMR (101 MHz, DMSO) σ 171.2, 159.7, 132.4, 119.3, 115.1 , 107.6, 79.2, 51.4, 43.6, 35.7, 28.0.

[0338] LRMS (ESI+, m / z) expected for Chemical Formula: C13H20N2O4 is 268.3 found 269.3 (M+H). tert-Butyl 4-(((allyloxy)carbonyl)amino)-1-(3-methoxy-3-oxopropyl)-1H-pyrrole-2- carboxylate

[0339] To a solution of terf-butyl 4-amino-1-(3-methoxy-3-oxopropyl)-1 / 7-pyrrole-2-carboxylate (0.890 g, 3.32 mmol, 1.0 eq) dissolved in DCM (DCM) (50 mL) in a flask flushed with argon and cooled to -78 °C was added N,N-diisopropylethylamine (643 mg, 867 uL, 4.98 mmol, 1 .5 eq) and allyl chloroformate (440 mg, 388 uL, 3.65 mmol, 1 .1 eq) consecutively, allowed to warm up to room temperature and stirred for 18 hours. The reaction was then quenched by the addition of water and the organic layer collected. The aqueous layer was extracted with DCM (2 x 20 mL) and the combined organic layers were concentrated and purified by flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 50%) to afford the target compound as an orange oil (1.01 g, 86%).

[0340] 1H NMR (400 MHz, DMSO) σ 9.41 (s, 1 H), 7.10 - 7.02 (m, 1 H), 6.63 (d, J = 2.1 Hz, 1 H), 6.02 - 5.89 (m, 1 H), 5.32 (dq, J = 17.2, 1.7 Hz, 1 H), 5.21 (dq, J = 10.5, 1.4 Hz, 1 H), 4.56 (dt, J = 5.5, 1.5 Hz, 2H), 4.43 (t, J = 6.9 Hz, 2H), 3.60 (s, 3H), 2.74 (t, J = 6.9 Hz, 2H), 1.49 (s, 9H).

[0341] 13C NMR (101 MHz, DMSO) σ 171.1 , 159.5, 153.1 , 133.5, 122.7, 119.7, 118.0, 117.3, 108.3, 80.0, 64.5, 51.4, 44.0, 35.6, 28.0.

[0342] 4-(((Allyloxy)carbonyl)amino)-1-(3-methoxy-3-oxopropyl)-1H-pyrrole-2-carboxylic acid

[0343] To a solution of terf-butyl 4-(((allyloxy)carbonyl)amino)-1-(3-methoxy-3-oxopropyl)-1 / 7- pyrrole-2-carboxylate (1.01 g, 2.87 mmol, 1.0 eq) in DCM (50.0 mL) in a sealed flask flushed with argon and cooled to -10 °C was added titanium (IV) chloride 1.0 M in DCM (5.94 mL, 34.4 mmol, 12.0 eq). The resultant mixture was stirred at this temperature for 2 hours then quenched with aqueous HCI (1M, 30 mL). The organic layer was collected, and the aqueous layer was extracted with DCM (2 x 30 mL). The combined organic layers were concentrated under reduced pressure and purified by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (30 to 100%) to afford the target compound as a light brown solid (0.628 g 74%)

[0344] 1H NMR (400 MHz, DMSO) σ 12.23 (s, 1 H), 9.43 (s, 1 H), 7.13 - 7.05 (m, 1 H), 6.66 (d, J = 2.0 Hz, 1 H), 5.96 (ddt, J = 17.2, 10.6, 5.4 Hz, 1 H), 5.32 (dq, J = 17.2, 1.7 Hz, 1 H), 5.21 (dq, J = 10.5, 1.5 Hz, 1 H), 4.57 (dq, J = 5.4, 2.2 Hz, 2H), 4.46 (t, J = 6.9 Hz, 2H), 3.59 (s, 3H), 2.75 (t, J = 6.8 Hz, 2H).

[0345] 13C NMR (101 MHz, DMSO) σ 171.2, 161.2, 153.1 , 133.5, 122.7, 119.1 , 118.0, 117.2, 108.3, 64.5, 51.4, 43.9, 35.6.

[0346] LRMS (ESI+, m / z) expected for Chemical Formula: C13H16N2O6 is 296.3 found 297.2 (M+H), 265.0 (M-H).

[0347] Route to 4-(((allyloxy)carbonyl)amino)-1-(2-(2-methoxyethoxy)ethyl)-1H-pyrrole-2- carboxylic acid tert-Butyl 1-(2-(2-methoxyethoxy)ethyl)-4-nitro-1H-pyrrole-2-carboxylate

[0348] To a mixture of terf-butyl 4-nitro-1 / 7-pyrrole-2-carboxylate (1.0 g, 4.71 mmol, 1.0 eq), 1- bromo-2-(2-methoxyethoxy)ethane (0.949 g, 0.698 mL, 5.18 mmol, 1.1 eq), potassium carbonate (0.977 g, 7.07 mmol, 1.5 eq) and tetrabutylammonium iodide (0.174 g, 0.471 mmol, 0.1 eq) in a sealed flask flushed with argon was added DMF (5 mL) and the resultant mixture stirred at 80 °C overnight. The reaction was then cooled and quenched by the addition of water and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (3 x 20 mL) and absorbed onto celite. Purification by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 40% percent) afforded the target compound as a yellow oil (1 .44 g, 97%).

[0349] 1H NMR (400 MHz, DMSO) σ 8.18 (d, J = 2.1 Hz, 1 H), 7.25 (d, J = 2.1 Hz, 1 H), 4.52 (t, J = 5.2 Hz, 2H), 3.75 - 3.68 (m, 2H), 3.55 - 3.49 (m, 2H), 3.41 - 3.36 (m, 2H), 3.21 (s, 3H), 1.52 (s, 9H).

[0350] 13C NMR (101 MHz, DMSO) σ 158.7, 134.1 , 129.1 , 123.4, 111.8, 81.8, 71.2, 69.4, 69.2, 58.0, 48.9, 27.7. tert-Butyl 4-amino-1-(2-(2-methoxyethoxy)ethyl)-1H-pyrrole-2-carboxylate

[0351] To a mixture of terf-butyl 1-(2-(2-methoxyethoxy)ethyl)-4-nitro-1 / 7-pyrrole-2-carboxylate (1.44 g, 4.57 mmol, 1 eq) and palladium on carbon 10% wt (0.210 g) in a 250 mL round bottom flask, sealed and flushed with argon was added methanol (50 mL) followed by a balloon of hydrogen and the resulting reaction mixture stirred at 45 °C overnight. On completion (monitored by LCMS), the crude reaction mixture was absorbed onto celite under reduced pressure and purified by flash chromatography eluting with a gradient of petroleum ether to 10% methanol in ethyl acetate (0 to 80%) to afford the target compound as a deep red oil (0.930 g, 72%).

[0352] 1H NMR (400 MHz, DMSO) σ 6.41 (d, J = 2.2 Hz, 1 H), 6.19 (d, J = 2.2 Hz, 1 H), 4.25 (t, J = 5.8 Hz, 2H), 3.86 (s, 2H), 3.59 (t, J = 5.8 Hz, 2H), 3.50 - 3.46 (m, 2H), 3.43 - 3.39 (m, 2H), 3.23 (s, 3H), 1.47 (s, 9H).

[0353] 13C NMR (101 MHz, DMSO) σ 159.8, 132.2, 119.5, 115.8, 107.5, 79.0, 71.2, 70.5, 69.5, 58.0, 47.3, 28.0.

[0354] LRMS (ESI+, m / z) expected for Chemical Formula: C14H24N2O4 is 284.4 found 285.2 (M+H). tert-Butyl 4-(((allyloxy)carbonyl)amino)-1-(2-(2-methoxyethoxy)ethyl)-1H-pyrrole- 2-carboxylate

[0355] To a solution of terf-butyl 4-amino-1-(2-(2-methoxyethoxy)ethyl)-1H-pyrrole-2- carboxylate (0.930 g, 3.27 mmol, 1.0 eq) dissolved in DCM (50 mL) in a flask flushed with argon and cooled to -78 °C, was added N,N-diisopropylethylamine (634 mg, 855 uL, 4.91 mmol, 1.5 eq) and allyl chloroformate (473 mg, 417 uL, 3.92 mmol, 1.2 eq) consecutively, allowed to warm up to room temperature and stirred for 18 hours. The reaction was then quenched by the addition of water and the organic layer collected. The aqueous layer was extracted with DCM (2 x 20 mL) and the combined organic layers were concentrated and purified by flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 50%) to afford the target compound as a dark oil (0.919 g, 76%).

[0356] 1H NMR (400 MHz, DMSO) σ 9.39 (s, 1 H), 7.07 (d, J = 2.1 Hz, 1 H), 6.64 (d, J = 2.0 Hz, 1 H), 5.96 (ddt, J = 17.3, 10.6, 5.4 Hz, 1 H), 5.32 (dq, J = 17.2, 1.7 Hz, 1 H), 5.22 (dq, J = 10.5, 1.4 Hz, 1 H), 4.56 (dt, J = 5.6, 1.5 Hz, 2H), 4.36 (t, J = 5.6 Hz, 2H), 3.62 (t, J = 5.5 Hz, 2H), 3.51 - 3.43 (m, 2H), 3.42 - 3.37 (m, 2H), 3.22 (s, 3H), 1.49 (s, 9H).

[0357] 13C NMR (101 MHz, DMSO) σ 159.7, 153.1 , 133.5, 122.5, 119.8, 118.5, 117.3, 108.1 , 79.8, 71.2, 70.3, 69.5, 64.5, 58.0, 47.8, 28.0, 28.0, 27.9.

[0358] LRMS (ESI+, m / z) expected for Chemical Formula: 368.4 C18H28N2O6 is found (M+H), (M-H).

[0359] 4-(((Allyloxy)carbonyl)amino)-1-(2-(2-methoxyethoxy)ethyl)-1H-pyrrole-2- carboxylic acid

[0360]

[0361] To a solution of terf-butyl 4-(((allyloxy)carbonyl)amino)-1-(2-(2-methoxyethoxy)ethyl)- 1 / 7-pyrrole-2-carboxylate (0.919 g, 2.49 mmol, 1.0 eq) in DCM (50.0 mL) in a sealed flask flushed with argon and cooled to -10 °C was added titanium (IV) chloride 1.0 M in DCM (5.17 mL, 29.9 mmol, 12.0 eq). The resultant mixture was stirred at this temperature for 2 hours then quenched with aqueous HCI (1M, 30 mL). The organic layer was collected, and the aqueous layer was extracted with DCM (2 x 30 mL). The combined organic layers were concentrated under reduced pressure and purified by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (30 to 100%) to afford the target compound as a light brown solid (0.546 g 70%)

[0362] 1H NMR (400 MHz, DMSO) σ 12.16 (s, 1 H), 9.42 (s, 1 H), 7.13 - 7.07 (m, 1 H), 6.66 (d, J = 2.0 Hz, 1 H), 5.96 (ddt, J = 17.2, 10.6, 5.4 Hz, 1 H), 5.33 (dq, J = 17.2, 1.7 Hz, 1 H), 5.22 (dq, J = 10.5, 1.5 Hz, 1 H), 4.61 - 4.54 (m, 2H), 4.39 (t, J = 5.6 Hz, 2H), 3.63 (t, J = 5.6 Hz, 2H), 3.50 - 3.46 (m, 2H), 3.42 - 3.38 (m, 2H), 3.22 (s, 3H).

[0363] 13C NMR (101 MHz, DMSO) σ 161.7, 153.1 , 133.5, 122.5, 119.1 , 118.6, 117.2, 108.2, 71.2, 70.3, 69.5, 64.5, 58.1 , 47.6.

[0364] LRMS (ESI+, m / z) expected for Chemical Formula: C14H20N2O6 is 312.3 found 313.2 (M+H), 311.1 (M-H).

[0365] Route to 4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-1-(2, 5, 8,11,14- pentaoxahexadecan-16-yl)-1H-pyrrole-2-carboxylic acid

[0366] tert-Butyl 1-(2,5,8,11,14-pentaoxahexadecan-16-yl)-4-nitro-1H-pyrrole-2- carboxylate

[0367] To a sealed flask containing terf-butyl 4-nitro-1 / 7-pyrrole-2-carboxylate (1.30 g, 6.13 mmol, 1.0 eq), tetrabutylammonium (226 mg, 6.13 umol, 0.1 eq), potassium carbonate (1.02 g, 7.34 mmol, 1.2 eq) flushed with argon was added DMF (5 mL) followed by 16- bromo-2,5,8,11,14-pentaoxahexadecane (1.93 g, 6.13 mmol, 1.0 eq) and the resulting mixture stirred at 80 °C overnight. The reaction was then cooled and quenched with water, diluted with EtOAc, washed with brine (3 x 10 mL) and the aqueous layer extracted with EtOAc (2 x 50 mL). The combined organic layers were absorbed onto celite and purified by automated flash silica chromatography eluting with a gradient of 0 to 100% EtOAc in petroleum ether to afford the desired product as a clear oil (2.45g, 90%).

[0368] 1H NMR (400 MHz, DMSO) σ 8.18 (d, J = 2.1 Hz, 1 H), 7.25 (d, J = 2.1 Hz, 1 H), 4.52 (t, J = 5.2 Hz, 2H), 3.75 - 3.70 (m, 2H), 3.48 (d, J = 5.9 Hz, 14H), 3.44 - 3.40 (m, 2H), 3.24 (s, 3H), 1.53 (s, 9H).

[0369] 13C NMR (101 MHz, DMSO) σ 158.7, 134.1 , 129.2, 129.1 , 123.4, 111.8, 111.8, 81.8, 71.2, 69.8, 69.7, 69.7, 69.5, 69.2, 58.0, 49.0, 27.7, 23.2. tert-Butyl 4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-1-(2,5,8,11,14- pentaoxahexadecan-16-y l)-1H-pyrrole-2-carboxylate

[0370] To a mixture of terf-butyl 1-(2,5,8,11 ,14-pentaoxahexadecan-16-yl)-4-nitro-1 / 7-pyrrole-2- carboxylate (2.4 g, 5.37 mmol, 1 eq) and palladium on carbon 10% wt (0.2.4 g) in a 100 mL round bottom flask, sealed and flushed with argon was added methanol (50 mL) followed by a balloon of hydrogen and the resulting reaction mixture stirred at 50 °C overnight. On completion (monitored by LCMS), the crude reaction mixture was filtered through a pad of celite and concentrated under reduced pressure afford the crude amine as a deep red oil. The crude product was then dissolved in DCM and Fmoc-cuccinimide (2.17g, 6.45 mmol, 1.2 eq) was added and the resultant mixture stirred at room temperature overnight. On completion (monitored by LCMS), the reaction mixture was absorbed onto celite and purified by automated flash silica chromatography eluting with a gradient of 20 to 100% EtOAc in petroleum ether to afford the desired product as a clear oil (0.85 g, 26%)

[0371] 1H NMR (400 MHz, DMSO) σ 9.45 (s, 1 H), 7.90 (d, J = 7.5 Hz, 2H), 7.73 (d, J = 7.4 Hz, 2H), 7.47 - 7.41 (m, 2H), 7.38 - 7.33 (m, 2H), 7.10 (d, J = 2.1 Hz, 1 H), 6.69 (d, J = 2.0 Hz, 1 H), 4.44 (d, J = 6.7 Hz, 2H), 4.37 (t, J = 5.6 Hz, 2H), 4.29 (t, J = 6.8 Hz, 1 H), 3.66 - 3.60 (m, 2H), 3.47 (d, J = 1.2 Hz, 16H), 3.41 - 3.39 (m, 2H), 3.22 (s, 3H), 1.50 (s, 9H).

[0372] 13C NMR (101 MHz, DMSO) σ 159.7, 153.3, 143.8, 140.8, 127.6, 127.1 , 125.0, 122.5, 120.1 , 119.8, 118.6, 108.2, 79.8, 71.2, 70.3, 69.8, 69.7, 69.7, 69.5, 65.4, 58.0, 47.8, 46.7, 28.0.

[0373] HRMS (ESI+, m / z) expected for: C35H46N2O9Exact Mass: 638.3203 found 639.3276 (M+H); 661.3090 (M+Na)

[0374] 4-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-1-(2,5,8,11,14- pentaoxahexadecan-16-y l)-1H-pyrrole-2-carboxylic acid

[0375] To a solution of terf-butyl 4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-1-(2,5,8,11 ,14- pentaoxahexadecan-16-yl)-1H-pyrrole-2-carboxylate (0.85 g, 1.33 mmol, 1.0 eq) in DCM cooled to -10°C in an ice / salt bath, TiCU (2.66 mmol, 2.0 eq as a 1M solution in DCM), was added dropwise and the resultant mixture stirred at -10°C for 1 hour. The reaction was then quenched by the addition of cold I M HCI. The organic layer was collected and the aqueous layer extracted with DCM (2 x 20 mL). The combined organic layers were dried (Na2SO4), absorbed onto celite and purified by automated flash chromatography eluting with a gradient of 0 to 100 % chloroform (10% MeOH in chloroform with 1 % acetic acid as buffer) to afford the desired product as an off white solid (acetic acid salt; 0.12 g, 16%).

[0376] 1H NMR (400 MHz, DMSO) σ 12.00 (s, 5H), 9.43 (s, 1 H), 7.91 (dt, J = 7.6, 1.0 Hz, 2H), 7.73 (d, J = 7.4 Hz, 2H), 7.42 (ddd, J = 7.5, 1 .2, 0.6 Hz, 2H), 7.35 (td, J = 7.4, 1 .2 Hz, 2H), 7.12 (d, J = 2.1 Hz, 1 H), 6.68 (d, J = 2.0 Hz, 1 H), 4.45 (d, J = 6.7 Hz, 2H), 4.43 - 4.36 (m, 2H), 4.28 (t, J = 6.8 Hz, 1 H), 3.63 (t, J = 5.6 Hz, 2H), 3.48 - 3.47 (m, 14H), 3.40 (d, J = 3.5 Hz, 2H), 3.22 (s, 3H).

[0377] 13C NMR (101 MHz, DMSO) σ 172.0, 161.7, 153.3, 143.8, 140.8, 127.6, 127.1 , 125.0, 122.5, 120.1 , 119.0, 118.7, 108.2, 71.2, 70.3, 69.8, 69.7, 69.6, 69.5, 65.4, 59.7, 58.0, 47.6, 46.7, 21.0.

[0378] LCMS (ESI+ m / z) 583.3 (10%) M+H, 600.3 (100%) M+18, (ESI - m / z) 581.2 (10%) M-H

[0379] Route to 4-(((allyloxy)carbonyl)amino)-1-(4-methoxy-4-oxobutyl)-1H-pyrrole-2- carboxylic acid tert-Butyl 1-(4-methoxy-4-oxobutyl)-4-nitro-1H-pyrrole-2-carboxylate

[0380] To a mixture of terf-butyl 4-nitro-1H-pyrrole-2-carboxylate (1.0 g, 4.71 mmol, 1.0 eq), methyl 4-bromobutyrate (0.938 g, 0.652 mL, 5.18 mmol, 1.1 eq), potassium carbonate (0.977 g, 7.07 mmol, 1.5 eq) and tetrabutylammonium iodide (0.174 g, 0.471 mmol, 0.1 eq) in a sealed flask flushed with argon was added DMF (5 mL) and the resultant mixture stirred at 80 °C overnight. The reaction was then cooled and quenched by the addition of water and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (3 x 20 mL) and absorbed onto celite. Purification by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 40% percent) afforded the target compound as a yellow oil (1.46 g, 99%).

[0381] 1H NMR (400 MHz, DMSO) σ 8.24 (d, J = 2.1 Hz, 1 H), 7.24 (d, J = 2.1 Hz, 1 H), 4.37 (t, J = 6.8 Hz, 2H), 3.56 (s, 3H), 2.31 (t, J = 7.4 Hz, 2H), 2.05 - 1.96 (m, 2H), 1.52 (s, 9H).

[0382] 13C NMR (101 MHz, DMSO) σ 172.4, 158.6, 134.3, 128.6, 123.3, 112.0, 81.7, 51.3, 48.7, 30.1 , 27.7, 25.8. tert-Butyl 4-amino-1-(4-methoxy-4-oxobutyl)-1H-pyrrole-2-carboxylate

[0383] To a mixture of tert-butyl 1-(4-methoxy-4-oxobutyl)-4-nitro-1 / 7-pyrrole-2-carboxylate (1.46 g, 4.67 mmol, 1 eq) and palladium on carbon 10% wt (0.230 g) in a 250 mL round bottom flask, sealed and flushed with argon was added methanol (50 mL) followed by a balloon of hydrogen and the resulting reaction mixture stirred at 45 °C overnight. On completion (monitored by LC-MS), the crude reaction mixture was absorbed onto celite under reduced pressure and purified by flash chromatography eluting with a gradient of petroleum ether to 10% methanol in ethyl acetate (0 to 80%) to afford the target compound as a deep red oil (0.93 g, 70%).

[0384] 1H NMR (400 MHz, DMSO) σ 6.37 (d, J = 2.3 Hz, 1 H), 6.19 (d, J = 2.2 Hz, 1 H), 4.12 (t, J = 6.8 Hz, 2H), 3.87 (s, 2H), 3.58 (s, 3H), 2.21 (t, J = 7.5 Hz, 2H), 1.87 (p, J = 7.2 Hz, 2H), 1.47 (s, 9H).

[0385] 13C NMR (101 MHz, DMSO) σ 172.7, 159.7, 132.3, 119.5, 115.2, 107.5, 79.0, 51.3, 46.6, 30.2, 28.0, 26.5.

[0386] LRMS (ESI+, m / z) expected for Chemical Formula: C14H22N2O4 is 282.3 found 283.3 (M+H). tert-Butyl 4-(((allyloxy)carbonyl)amino)-1-(4-methoxy-4-oxobutyl)-1H-pyrrole-2- carboxylate

[0387]

[0388] To a solution of terf-butyl 4-amino-1-(4-methoxy-4-oxobutyl)-1 / 7-pyrrole-2-carboxylate (0.930 g, 3.29 mmol, 1.0 eq) dissolved in DCM (50 mL) in a flask flushed with argon and cooled to -78 °C was added N,N-diisopropylethylamine (639 mg, 861 uL, 4.94 mmol, 1.5 eq) and allyl chloroformate (476 mg, 420 uL, 3.95 mmol, 1.2 eq) consecutively, allowed to warm up to room temperature and stirred for 18 hours. The reaction was then quenched by the addition of water and the organic layer collected. The aqueous layer was extracted with DCM (2 x 20 mL) and the combined organic layers were concentrated and purified by flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 50%) to afford the target compound as a red oil (1.15 g, 95%).

[0389] 1H NMR (400 MHz, DMSO) σ 9.41 (s, 1 H), 7.04 (d, J = 2.1 Hz, 1 H), 6.62 (d, J = 2.0 Hz, 1 H), 5.32 (dq, J = 17.2, 1.7 Hz, 1 H), 5.22 (dq, J = 10.4, 1.4 Hz, 1 H), 4.56 (dt, J = 5.6, 1.5 Hz, 2H), 4.22 (t, J = 6.8 Hz, 2H), 3.58 (s, 3H), 2.23 (t, J = 7.5 Hz, 2H), 1.90 (p, J = 7.2 Hz, 2H), 1.49 (s, 9H).

[0390] 13C NMR (101 MHz, DMSO) σ 172.6, 159.6, 153.1 , 133.5, 122.6, 119.8, 117.9, 117.3, 108.1 , 79.8, 64.5, 51.3, 47.1 , 30.2, 28.0, 26.5.

[0391] 4-(((Allyloxy)carbonyl)amino)-1-(4-methoxy-4-oxobutyl)-1H-pyrrole-2-carboxylic acid

[0392] To a solution of terf-butyl 4-(((allyloxy)carbonyl)amino)-1-(4-methoxy-4-oxobutyl)-1 / 7- pyrrole-2-carboxylate (1.15 g, 3.14 mmol, 1.0 eq) in DCM (50.0 mL) in a sealed flask flushed with argon and cooled to -10 °C was added titanium (IV) chloride 1.0 M in DCM (6.50 mL, 37.7 mmol, 12.0 eq). The resultant mixture was stirred at this temperature for 2 hours then quenched with aqueous HCI (1M, 30 mL). The organic layer was collected, and the aqueous layer was extracted with DCM (2 x 30 mL). The combined organic layers were concentrated under reduced pressure and purified by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (30 to 100%) to afford the target compound as a light brown solid (0.841 g 86%)

[0393] 1H NMR (400 MHz, DMSO) σ 12.16 (s, 1 H), 9.44 (s, 1 H), 7.14 - 7.02 (m, 1 H), 6.65 (d, J = 2.0 Hz, 1 H), 5.96 (ddt, J = 17.2, 10.6, 5.4 Hz, 1 H), 5.33 (dq, J = 17.3, 1.8 Hz, 1 H), 5.22 (dq, J = 10.4, 1.4 Hz, 1 H), 4.57 (dt, J = 5.5, 1.5 Hz, 2H), 4.25 (t, J = 6.8 Hz, 2H), 3.57 (s, 3H), 2.22 (t, J = 7.5 Hz, 2H), 1.91 (p, J = 7.2 Hz, 2H).

[0394] 13C NMR (101 MHz, DMSO) σ 172.7, 161.6, 153.1 , 133.5, 122.6, 119.2, 117.9, 117.3, 108.1 , 64.5, 51.3, 47.0, 30.2, 26.4.

[0395] LRMS (ESI+, m / z) expected for Chemical Formula: CuHwNzOe is 310.3 found

[0396] 311.2 (M+H), 309.1 (M-H).

[0397] Route to 4-(((allyloxy)carbonyl)amino)-1-(3-cyanopropyl)-1H-pyrrole-2-carboxylic acid tert-Butyl 1-(3-cyanopropyl)-4-nitro-1H-pyrrole-2-carboxylate

[0398] To a mixture of terf-butyl 4-nitro-1 / 7-pyrrole-2-carboxylate (1.0 g, 4.71 mmol, 1.0 eq), 4- bromobutanenitrile (0.837 g, 5.65 mmol, 1.2 eq), potassium carbonate (0.977 g, 7.07 mmol, 1.5 eq) and tetrabutylammonium iodide (0.174 g, 0.471 mmol, 0.1 eq) in a sealed flask flushed with argon was added DMF (5 mL) and the resultant mixture stirred at 80 °C overnight. The reaction was then cooled and quenched by the addition of water and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (3 x 20 mL) and absorbed onto celite. Purification by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 40% percent) afforded the target compound as a yellow oil (1.32 g, 100%).

[0399] 1H NMR (400 MHz, DMSO) σ 8.27 (d, J = 2.1 Hz, 1 H), 7.27 (d, J = 2.1 Hz, 1 H), 4.41 (dd, J = 7.5, 6.4 Hz, 2H), 2.54 (t, J = 7.1 Hz, 2H), 2.07 (p, J = 7.2 Hz, 2H), 1.53 (s, 9H).

[0400] 13C NMR (101 MHz, DMSO) σ 158.6, 134.4, 128.7, 123.4, 119.7, 112.1 , 81.9, 48.3, 27.7, 26.2, 13.6.

[0401] LRMS (ESI+, m / z) expected for Chemical Formula: C13H17N3O4is 279.3 found 283.1 (M+H), 281.1 (M-H). tert-Butyl 4-amino-1-(3-cyanopropyl)-1H-pyrrole-2-carboxylate

[0402] To a mixture of tert-butyl 1-(3-cyanopropyl)-4-nitro-1H-pyrrole-2-carboxylate (1.28 g, 4.58 mmol, 1 eq) and palladium on carbon 10% wt (0.120 g) in a 250 mL round bottom flask, sealed and flushed with argon was added methanol (50 mL) followed by a balloon of hydrogen and the resulting reaction mixture stirred at 45 °C overnight. On completion (monitored by LC-MS), the crude reaction mixture was absorbed onto celite under reduced pressure and purified by flash chromatography eluting with a gradient of petroleum ether to 10% methanol in ethyl acetate (0 to 80%) to afford the target compound as a deep red oil (0.992 g, 87%).

[0403] 1H NMR (500 MHz, DMSO) σ 6.40 (d, J = 2.3 Hz, 1 H), 6.21 (d, J = 2.2 Hz, 1 H), 4.16 (t, J = 6.9 Hz, 2H), 3.92 (s, 2H), 2.40 (t, J = 7.2 Hz, 2H), 1 .92 (p, J = 7.2 Hz, 2H), 1 .48 (s, 9H).

[0404] LRMS (ESI+, m / z) 250.3 M+ H, 20% tert-Butyl 4-(((allyloxy)carbonyl)amino)-1-(3-cyanopropyl)-1H-pyrrole-2- carboxylate

[0405] To a solution of terf-butyl 4-amino-1-(3-cyanopropyl)-1 / 7-pyrrole-2-carboxylate (0.992 g, 3.98 mmol, 1.0 eq) dissolved in DCM (50 mL) in a flask flushed with argon and cooled to -78 °C was added N,N-diisopropylethylamine (771 mg, 1.04 mL, 5.97 mmol, 1.5 eq) and allyl chloroformate (576 mg, 508 uL, 4.77 mmol, 1.2 eq) consecutively, allowed to warm up to room temperature and stirred for 18 hours. The reaction was then quenched by the addition of water and the organic layer collected. The aqueous layer was extracted with DCM (2 x 20 mL) and the combined organic layers were concentrated and purified by flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 50%) to afford the target compound as a red oil (0.998 g, 75%).

[0406] 1H NMR (400 MHz, DMSO) σ 9.44 (s, 1 H), 7.14 - 7.06 (m, 1 H), 6.64 (d, J = 2.0 Hz, 1 H), 5.96 (ddt, J = 17.0, 10.5, 5.4 Hz, 1 H), 5.37 - 5.29 (m, 1 H), 5.22 (dq, J = 10.5, 1.5 Hz, 1 H), 4.57 (dt, J = 5.5, 1.5 Hz, 2H), 4.26 (dd, J = 7.7, 6.3 Hz, 2H), 2.43 (t, J = 7.2 Hz, 2H), 1.95 (p, J = 7.2 Hz, 2H), 1.50 (s, 9H).

[0407] 13C NMR (101 MHz, DMSO) σ 159.6, 153.1 , 133.5, 122.8, 119.9, 119.8, 117.8, 117.3, 108.3, 80.0, 64.5, 46.9, 28.0, 26.8, 13.6.

[0408] LRMS (ESI+, m / z) expected for Chemical Formula: C17H23N3O4 is 333.4 found 334.3 (M+H), 332.2 (M-H).

[0409] 4-(((Allyloxy)carbonyl)amino)-1-(3-cyanopropyl)-1H-pyrrole-2-carboxylic acid

[0410] To a solution of terf-butyl 4-(((allyloxy)carbonyl)amino)-1-(3-cyanopropyl)-1 / 7-pyrrole-2- carboxylate (998 mg, 2.99 mmol, 1.0 eq) in DCM (50.0 mL) in a sealed flask flushed with argon and cooled to -10 °C was added titanium (IV) chloride 1.0 M in DCM (6.20 mL, 35.9 mmol, 12.0 eq). The resultant mixture was stirred at this temperature for 2 hours then quenched with aqueous HCI (1M, 30 mL). The organic layer was collected, and the aqueous layer was extracted with DCM (2 x 30 mL). The combined organic layers were concentrated under reduced pressure and purified by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (30 to 100%) to afford the target compound as a light brown solid (0.603 g 73%)

[0411] 1H NMR (400 MHz, DMSO) σ 12.24 (s, 1 H), 9.46 (s, 1 H), 7.18 - 7.09 (m, 1 H), 6.68 (d, J = 2.0 Hz, 1 H), 5.96 (ddt, J = 17.3, 10.7, 5.4 Hz, 1 H), 5.33 (dq, J = 17.3, 1.7 Hz, 1 H), 5.21 (dq, J = 10.4, 1.4 Hz, 1 H), 4.57 (dt, J = 5.4, 1.5 Hz, 2H), 4.35 - 4.26 (m, 2H), 2.42 (t, J = 7.2 Hz, 2H), 1.97 (p, J = 7.1 Hz, 2H).

[0412] 13C NMR (101 MHz, DMSO) σ 161.6, 153.1 , 133.4, 122.8, 119.9, 119.3, 117.9, 117.3, 108.3, 64.5, 46.7, 26.8, 13.6.

[0413] LRMS (ESI+, m / z) expected for Chemical Formula: C13H15N3O4 is 277.3 found 278.2 (M+H), 277.2 (M-H).

[0414] Route to 4-(((allyloxy)carbonyl)amino)-1-(3-methoxypropyl)-1H-pyrrole-2- carboxylic acid tert-Butyl 1-(3-methoxypropyl)-4-nitro-1H-pyrrole-2-carboxylate

[0415] To a mixture of terf-butyl 4-nitro-1 / 7-pyrrole-2-carboxylate (1.0 g, 4.71 mmol, 1.0 eq),1- bromo-3-methoxypropane (0.793 g, 0.583 mL, 5.18 mmol, 1.1 eq) mmol, potassium carbonate (0.977 g, 7.07 mmol, 1.5 eq) and tetrabutylammonium iodide (0.174 g, 0.471 mmol, 0.1 eq) in a sealed flask flushed with argon was added dimethlyformamide (5 mL) and the resultant mixture stirred at 80 °C overnight. The reaction was then cooled and quenched by the addition of water and extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (3 x 20 mL) and absorbed onto celite. Purification by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 40% percent) afforded the target compound as a yellow oil (1.46 g, 99%).

[0416] 1H NMR (400 MHz, DMSO) σ 8.24 (d, J = 2.1 Hz, 1 H), 7.24 (d, J = 2.1 Hz, 1 H), 4.37 (t, J = 6.8 Hz, 2H), 3.56 (s, 3H), 2.31 (t, J = 7.4 Hz, 2H), 2.00 (p, J = 7.1 Hz, 2H), 1.53 (s, 9H). tert-Butyl 4-amino-1-(3-methoxypropyl)-1H-pyrrole-2-carboxylate

[0417] To a mixture of terf-butyl 1-(3-methoxy-3-oxopropyl)-4-nitro-1 / 7-pyrrole-2-carboxylate (1.32 g, 4.64 mmol, 1 eq) and palladium on carbon 10% wt (0.220 g) in a 250 mL round bottom flask, sealed and flushed with argon was added methanol (50 mL) followed by a balloon of hydrogen and the resulting reaction mixture stirred at 45 °C overnight. On completion (monitored by LC-MS), the crude reaction mixture was absorbed onto celite under reduced pressure and purified by flash chromatography eluting with a gradient of petroleum ether to 10% methanol in ethyl acetate (0 to 80%) to afford the target compound as a deep red oil (1.06 g, 90%).

[0418] 1H NMR (400 MHz, DMSO) σ 6.35 (d, J = 2.2 Hz, 1 H), 6.19 (d, J = 2.2 Hz, 1 H), 4.12 (t, J = 7.0 Hz, 2H), 3.85 (s, 2H), 3.23 (d, J = 5.9 Hz, 5H), 1.87 - 1.78 (m, 2H), 1.48 (s, 9H).13C NMR (101 MHz, DMSO) σ 159.7, 132.2, 119.4, 115.3, 107.4, 79.0, 68.9, 57.8, 44.9, 31.2, 28.0.

[0419] LRMS (ESI+, m / z) expected for Chemical Formula: C13H22N2O3 is 254.3 found 255.3 (M+H). tert-Butyl 4-(((allyloxy)carbonyl)amino)-1-(3-methoxypropyl)-1H-pyrrole-2- carboxylate

[0420] To a solution of terf-butyl 4-amino-1-(3-methoxypropyl)-1 / 7-pyrrole-2-carboxylate (1.06 g, 4.17 mmol, 1.0 eq) dissolved in DCM (50 mL) in a flask flushed with argon and cooled to -78 °C, was added N,N-diisopropylethylamine (808 mg, 1.09 mL, 6.25 mmol, 1.5 eq) and allyl chloroformate (603 mg, 532 uL, 5.0 mmol, 1.2 eq) consecutively, allowed to warm up to room temperature and stirred for 18 hours. The reaction was then quenched by the addition of water and the organic layer collected. The aqueous layer was extracted with DCM (2 x 20 mL) and the combined organic layers were concentrated and purified by flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 50%) to afford the target compound as a dark red oil (1.17 g, 83%).

[0421] 1H NMR (400 MHz, DMSO) σ 9.40 (s, 1 H), 7.02 (d, J = 2.0 Hz, 1 H), 6.63 (d, J = 2.1 Hz, 1 H), 6.02 - 5.89 (m, 1 H), 5.32 (dq, J = 17.3, 1.7 Hz, 1 H), 5.21 (dq, J = 10.5, 1.5 Hz, 1 H), 4.56 (dd, J = 5.6, 1.7 Hz, 2H), 4.23 (t, J = 6.9 Hz, 2H), 3.23 (d, J = 4.0 Hz, 5H), 1.91 - 1.80 (m, 2H), 1.49 (s, 9H).

[0422] 13C NMR (101 MHz, DMSO) σ 159.6, 153.1 , 133.5, 122.5, 119.8, 118.0, 117.3, 108.1 , 79.7, 79.7, 68.6, 68.6, 64.5, 57.8, 45.3, 31.0, 27.9. 339.3 LRMS (ESI+, m / z) (M+H, 60%), 337.2 (M-H, 20%)

[0423] LRMS (ESI+, m / z) expected for Chemical Formula: C17H26N2O5 is 338.4 found (M+H), (M-H).

[0424] 4-(((Allyloxy)carbonyl)amino)-1-(3-methoxypropyl)-1H-pyrrole-2-carboxylic acid To a solution of terf-butyl 4-(((allyloxy)carbonyl)amino)-1-(3-methoxypropyl)-1 / 7-pyrrole- 2-carboxylate (1.17 g, 3.46 mmol, 1.0 eq) in DCM (50.0 mL) in a sealed flask flushed with argon and cooled to -10 °C was added titanium (IV) chloride 1.0 M in DCM (7.16 mL, 41.5 mmol, 12.0 eq). The resultant mixture was stirred at this temperature for 2 hours then quenched with aqueous HCI (1M, 30 mL). The organic layer was collected, and the aqueous layer was extracted with DCM (2 x 30 mL). The combined organic layers were concentrated under reduced pressure and purified by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (30 to 100%) to afford the target compound as a light brown solid (0.675 g 69%)

[0425] 1H NMR (400 MHz, DMSO) σ 9.42 (s, 1 H), 7.07 - 7.01 (m, 1 H), 6.64 (d, J = 2.0 Hz, 1 H), 5.96 (ddt, J = 17.3, 10.6, 5.4 Hz, 1 H), 5.33 (dq, J = 17.2, 1.7 Hz, 1 H), 5.22 (dq, J = 10.5, 1.4 Hz, 1 H), 4.56 (d, J = 5.3 Hz, 2H), 4.26 (t, J = 6.9 Hz, 2H), 3.23 (d, J = 4.1 Hz, 5H), 1.86 (p, J = 6.5 Hz, 2H).

[0426] 13C NMR (101 MHz, DMSO) σ 161.6, 153.1 , 133.5, 122.5, 118.0, 117.3, 113.9, 108.0, 68.7, 64.5, 57.8, 45.2, 30.9.

[0427] LRMS (ESI+, m / z) expected for Chemical Formula: C13H18N2O5is 282.3 found 283.1 (M+H), 281.1 (M-H).

[0428] Route to 4-(((allyloxy)carbonyl)amino)-1-(2-((triisopropylsilyl)oxy)ethyl)-1 H- pyrrole-2-carboxylic acid tert-Butyl 4-nitro-1-(2-((triisopropylsilyl)oxy)ethyl)-1H-pyrrole-2-carboxylate

[0429] To a mixture of terf-butyl 4-nitro-1 / 7-pyrrole-2-carboxylate (1.0 g, 4.71 mmol, 1.0 eq), (2- bromoethoxy)triisopropylsilane (1.46 g, 5.18 mmol, 1.1 eq) mmol, potassium carbonate (0.977 g, 7.07 mmol, 1.5 eq) and tetrabutylammonium iodide (0.174 g, 0.471 mmOI, 0.1 eq) in a sealed flask flushed with argon was added DMF (5 mL) and the resultant mixture stirred at 80 °covernight. The reaction was then cooled and quenched by the addition of water and extracted with ethyl acetate (3 X 50 mL). The combined organic layers were washed with brine (3 X 20 mL) and absorbed onto celite. Purification by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 40% percent) afforded the target compound as a yellow oil (1 .5 g, 77%).

[0430] 1H NMR (500 MHz, DMSO) σ 8.17 (d, J = 2.1 Hz, 1 H), 7.26 (d, J = 2.0 Hz, 1 H), 4.50 (t, J = 5.1 Hz, 2H), 3.95 (t, J = 5.1 Hz, 2H), 1.52 (s, 9H), 1.03 - 0.91 (m, 21 H).

[0431] 13C NMR (101 MHz, DMSO) σ 158.7, 134.0, 129.4, 123.1 , 111.7, 81.3, 62.0, 51.4, 27.5, 17.4, 11.2.

[0432] LRMS (ESI+, m / z) 453 (M+41 , 10%) tert-Butyl 4-(((allyloxy)carbonyl)amino)-1-(2-((triisopropylsilyl)oxy)ethyl)-1 H- pyrrole-2-carboxylate

[0433] To a solution of terf-butyl 4-nitro-1-(2-((triisopropylsilyl)oxy)ethyl)-1H-pyrrole-2- carboxylate (1.5 g, 3.64 mmol) in methanol (50mL) was added palladium on carbon 10% wt (0.165 g) in a 250 mL round bottom flask, sealed and flushed with argon. A balloon of hydrogen was then fitted, and the resulting reaction mixture stirred at 45 °covernight. On completion (monitored by LCMS), the crude reaction mixture was filtered and concentrated under reduced pressure. The crude product, terf-butyl 4-amino-1-(2- ((triisopropylsilyl)oxy)ethyl)-1 / 7-pyrrole-2-carboxylate was redissolved in DOM (50 mL) in a 100mL flask, flushed with argon and cooled to -10 °C. N,N-Diisopropylethylamine (659 mg, 496 uL, 5.10 mmol, 1.50 eq) and allyl chloroformate (491 mg, 433 uL, 4.08 mmol, 1.2 eq) were added consecutively, and the reaction was allowed to warm up to room temperature and stirred for 18 hours. The reaction was then quenched by the addition of water and the organic layer collected. The aqueous layer was extracted with DCM (2 x 20 mL) and the combined organic layers were concentrated and purified by flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (0 to 50%) to afford the target compound as a dark red oil (0.860 g, 54%).

[0434] 1H NMR (400 MHz, DMSO) σ 9.35 (s, 1 H), 7.06 (d, J = 2.2 Hz, 1 H), 6.63 (d, J = 2.0 Hz, 1 H), 5.94 (ddd, J = 17.2, 10.6, 5.3 Hz, 1 H), 5.31 (dq, J = 17.2, 1.7 Hz, 1 H), 5.20 (dq, J = 10.5, 1.4 Hz, 1 H), 4.55 (dt, J = 5.4, 1.6 Hz, 2H), 4.31 (t, J = 5.5 Hz, 2H), 3.86 (t, J = 5.4 Hz, 2H), 1 .47 (s, 10H), 0.96 - 0.93 (m, 21 H).

[0435] 13C NMR (101 MHz, DMSO) σ 159.8, 153.0, 133.5, 122.4, 119.6, 118.8, 117.1 , 108.0, 79.5, 64.4, 63.1 , 50.4, 27.9, 17.6, 11.3.

[0436] LRMS (ESI+, m / z) 467 (M+H, 100%), 465(M-H, 90%).

[0437] 4-(((Allyloxy)carbonyl)amino)-1-(2-((triisopropylsilyl)oxy)ethyl)-1 / 7-pyrrole-2- carboxylic acid

[0438] To a solution of terf-butyl 4-(((allyloxy)carbonyl)amino)-1-(2-((triisopropylsilyl)oxy)ethyl)-

[0439] 1 H-pyrrole-2-carboxylate (0.919 g, 2.49 mmol, 1 .0 eq) in DCM (50.0 mL) in a sealed flask flushed with argon and cooled to -10 oC was added Titanium (IV) Chloride 1 .0 M in DCM (5.17 mL, 29.9 mmol, 12.0 eq). The resultant mixture was stirred at this temperature for

[0440] 2 hours then quenched with HCI (1M, 30 mL). The organic layer was collected and the aqueous layer was extracted with DCM (2 x 30 mL). The combined organic layers were concentrated under reduced pressure and purified by automated flash chromatography eluting with a gradient of ethyl acetate in petroleum ether (30 to 100%) to afford the target compound as a light brown solid (0.524 g 60%)

[0441] 1H NMR (400 MHz, DMSO) σ 12.09 (s, 1 H), 9.39 (s, 1 H), 7.10 (d, J = 2.1 Hz, 1 H), 6.65 (d, J = 2.0 Hz, 1 H), 5.95 (ddt, J = 17.2, 10.6, 5.4 Hz, 1 H), 5.32 (dq, J = 17.3, 1.7 Hz, 1 H), 5.21 (dq, J = 10.5, 1.5 Hz, 1 H), 4.55 (d, J = 5.4 Hz, 2H), 4.35 (t, J = 5.5 Hz, 2H), 3.87 (t, J = 5.5 Hz, 2H), 1.05 - 0.94 (m, 21 H).

[0442] LRMS (ESI+, m / z) 411.3 (M+H, 100%), 409.3 (M-H, 100%), 819.4 (2M-H, 10%) Route to 5-(3-((((9H-fluoren-9- yl)methoxy)carbonyl)amino)propanamido)thiophene-2-carboxylic acid

[0443] FmocHN tert-Butyl 5-nitrothiophene-2-carboxylate

[0444] To a stirred solution of 5-nitrothiophene-2-carboxylic acid (866 mg, 5.00 mmol, 1.0 eq) in DCM (10.0 mL, 0.50 M) at RT, was added oxalyl chloride (645 pL, 7.50 mmol, 1.5 eq) and DMF (ca. 20 drops, catalytic). The resulting mixture stirred at RT for 30 min and concentrated in vacuo. To the residue was added a 1 :1 mixture of pyridine and tert butanol (10 mL) at RT and the reaction was stired for 1 hour. The mixture was partitioned between ethyl acetate (30 mL) and water (30 mL) and the organic phase separated. The aqueous phase was extracted with ethyl acetate (3 X 10 mL) and the combined organic phase was dried (MgSCU), filtered, and concentrated in vacuo to afford the title compound as a beige solid (993 mg, 4.33 mmol, 87%) which was used further without purification.

[0445] 1H NMR (400 MHz, DMSO-d6) δ 8.11 (d, J = 4.4 Hz, 1 H), 7.72 (d, J = 4.4 Hz, 1 H), 1.55 (s, 9H).

[0446] 13C NMR (101 MHz, DMSO-d6) δ 159.2, 154.0, 139.8, 132.1 , 129.7, 83.7, 27.6. tert-Butyl 5-aminothiophene-2-carboxylate

[0447] To round bottomed flask containing tert-butyl 5-nitrothiophene-2-carboxylate (928 mg, 4.04 mmol, 1.0 equiv.) and palladium on charcoal [10 wt. %] (431 mg, 0.40 mmol, 0.1 equiv.) under an inert atmosphere was added anhydrous methanol (20 mL, 0.20 M). An atmosphere of hydrogen gas was applied, and the resulting mixture was stirred at RT for 6 hours. The mixture was filtered over celite and concentrated in vacuo to afford the title compound as a beige solid (746 mg, 3.74 mmol, 93%) which was used further without purification.

[0448] 1H NMR (400 MHz, DMSO-d6) δ 7.23 (d, J = 4.1 Hz, 1 H), 6.55 (br s, 2H), 5.85 (d, J = 4.1 Hz, 1 H), 1.45 (s, 9H).13C NMR (101 MHz, DMSO-d6) δ 162.2, 161.4, 134.7, 114.0, 104.2, 79.3, 28.1. tert-Butyl 5-(3-((((9H-fluoren-9- yl)methoxy)carbonyl)amino)propanamido)thiophene-2-carboxylate

[0449] FmocHN

[0450] Fmoc-β-Ala-OH (1.26 g, 4.03 mmol), 4-dimethylaminopyridine (493 mg, 4.03 mmol, 1.2 eq) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (773 mg, 4.03 mmol, 1.2 eq) were dissolved in DCM (16.8 mL, 0.20 M) and the resulting mixture stirred at RT for 5 min. Tert-butyl 5-aminothiophene-2-carboxylate (670 mg, 3.36 mmol, 1 eq) was added and the resulting mixture stirred at rt for 16h. Aqueous HCI (1M) was added and the layers were separated. The aqueous was extracted with DCM (3 X 20mL) and the combined organic layers dried with Na2SC>4 and concentrated in vacuo. Purification by flash column chromatography (silica gel, 0-100% EtOAc / Hexane) afforded the desired product as a white solid (604 mg, 36%).

[0451] 1H NMR (400 MHz, DMSO-d6) 6 11.58 (s, 1 H), 7.88 (dt, J = 7.6, 1.0 Hz, 2H), 7.67 (d, J = 7.4 Hz, 2H), 7.48 (d, J = 4.1 Hz, 1 H), 7.46 - 7.43 (m, 1 H), 7.40 (td, J = 7.5, 1.1 Hz, 2H), 7.30 (td, J = 7.5, 1.3 Hz, 2H), 6.63 (d, J = 4.2 Hz, 1 H), 4.28 (d, J = 7.0 Hz, 2H), 4.21 (d, J = 6.9 Hz, 1 H), 3.33 - 3.28 (m, 2H), 2.57 (t, J = 6.8 Hz, 2H), 1 .50 (s, 9H).

[0452] 13C NMR (101 MHz, DMSO-d6) δ 168.2, 161.6, 156.1 , 145.7, 143.9, 140.7, 131.3, 127.6, 127.0, 125.1 , 123.7, 120.1 , 111.2, 80.5, 65.4, 46.7, 36.5, 35.4, 27.9.

[0453] 5-(3-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)propanamido)thiophene-2- carboxylic acid

[0454] FmocHN

[0455] 5-(3-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)propanamido)thiophene-2-carboxylic acid (600 mg, 1.22 mmol, 1.0 eq) was dissolved in DOM (10.0 mL, 0.12 M) and cooled to 0 °C. Trifluoroacetic acid (93.3 uL, 1.22 mmol, 1 eq) was added and the reaction mixture stirred at RT for 2h. The mixture was concentrated in vacuo and purified by flash column chromatography (silica gel, 0-100% EtOAc / Hexane) afforded the desired product as a beige / brown / grey solid (340 mg, 64%).1H NMR (400 MHz, DMSO-d6) δ 12.56 (s, 1 H), 11.57 (s, 1 H), 7.88 (dt, J = 7.6, 1.0 Hz, 2H), 7.67 (d, J = 7.5 Hz, 2H), 7.51 (d, J = 4.1 Hz, 1 H), 7.46 (t, J = 5.7 Hz, 1 H), 7.40 (td, J = 7.5, 1.2 Hz, 2H), 7.30 (td, J = 7.4, 1 .2 Hz, 2H), 6.65 (d, J = 4.2 Hz, 1 H), 4.29 (d, J = 7.1 Hz, 2H), 4.21 (t, J = 6.9 Hz, 1 H), 3.33 - 3.28 (m, 2H), 2.57 (t, J = 6.8 Hz, 2H).

[0456] 13C NMR (101 MHz, DMSO-d6) δ 168.2, 163.6, 156.1 , 145.9, 143.9, 140.7, 131.4, 127.6, 127.0, 125.1 , 123.4, 120.1 , 111.4, 65.4, 46.7, 36.5, 35.4.

[0457] Route to 3-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-bromo- 1-methyl-1H-pyrazole-5-carboxylic acid

[0458] Methyl 1-methyl-3-nitro-1H-pyrazole-5-carboxylate

[0459] A oven-dried round bottom flask was charged with methyl 5-nitro-1H-pyrazole-3- carboxylate (1.91 g, 11.2 mmol, 1.0 eq) and potassium carbonate (4.63 g, 33.5 mmol, 3.0 eq) then sealed and purged with nitrogen. The reactants were suspended in DMF (30 mL, 0.37 M) and the reaction was set to stir at RT before adding iodomethane (1 .04 mL, 16.7 mmol, 1.5 eq) . The resulting mixture was stirred at RT for 18 h. The reaction mixture was then diluted with 5% aqueous LiCI (50 mL) and extracted with DCM (3 x 60 mL). The combined organics were washed with brine (150 mL), dried over Na2SO4 and evaporated in vacuo to afford the crude product. Purification by flash column chromatography (silica gel, 0-20% EtOAc / Hexane) afforded the desired product as a white solid (consistent with a 86:14 mixture of regioisomers) (1.81 g, 88%).

[0460] 1H NMR (400 MHz, CDCI3) δ 7.39 (s, 1 H), 4.29 (s, 3H), 3.95 (s, 3H).13C NMR (101 MHz, CDCI3) δ 158.8, 134.8, 108.6, 107.2, 52.9, 41.2.

[0461] Methyl 3-amino-1-methyl-1H-pyrazole-5-carboxylate A round bottomed flask was charged with methyl 1-methyl-3-nitro-1 / 7-pyrazole-5- carboxylate (200 mg, 1.08 mmol, 1 eq) and Pd / C (115 mg, 108 pmol, 10 mol%) and the flask was purged with nitrogen. MeOH (10.8 mL, 0.1 M) was added and the reaction mixture was bubbled with H2for 30 min, then stirred under a H2atmosphere for 18 h. The reaction mixture was diluted with Et2O (20 mL) and filtered through celite, then concentrated in vacuo to afford the crude product as a white solid (regioisomer mixture). Purification by flash column chromatography (silica gel, 0-40% EtOAc / Hexane) afforded the desired product as a white solid and single regioisomer (130 mg, 78%).

[0462] 1H NMR (400 MHz, CDCI3) 5 6.19 (s, 1 H), 4.01 (s, 3H), 3.85 (s, 3H), 3.36 (br s, 2H).13C NMR (101 MHz, CDCI3) 5 160.3, 151.6, 133.1 , 98.0, 52.0, 38.7.

[0463] Methyl 3-amino-4-bromo-1-methyl-1 / 7-pyrazole-5-carboxylate

[0464] To a stirred solution of methyl 3-amino-1-methyl-1 / 7-pyrazole-5-carboxylate (774 mg, 4.99 mmol, 1 .0 equiv.) in THF (20.0 mL, 0.25 M) at 0 °C, was added N-bromosuccinimide (977 mg, 5.49 mmol, 1.1 equiv.) and the resulting mixture stirred at RT for 30 min. The reaction mixture was filtered through celite, then concentrated in vacuo. Purification by flash column chromatography (silica gel, 0-80% EtOAc / Hexane) afforded the desired product as a yellow solid (1.10 g, 95%).

[0465] 1H NMR (400 MHz, CDCI3) δ 4.00 (s, 3H), 3.92 (s, 3H), 3.47 (br s, 2H).

[0466] 13C NMR (101 MHz, CDCI3) δ 159.6, 151.7, 130.4, 85.4, 52.2, 40.2.

[0467] Methyl 3-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-bromo- 1-methyl-1H-pyrazole-5-carboxylate

[0468] Fmoc-β-Ala-OH (299 mg, 0.96 mmol, 1.2 eq), 4-dimethylaminopyridine (117 mg, 0.96 mmol, 1.2 eq) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (184 mg, 0.96 mmol, 1.2 eq) were dissolved in DCM (4.0 mL, 0.20 M) and the resulting mixture stirred at RT for 5 min. Methyl 3-amino-4-bromo-1-methyl-1H-pyrazole-5- carboxylate (187 mg, 0.80 mmol, 1 eq) was added and the resulting mixture stirred at rt for 16h. Aqueous HCI (1M) was added and the layers were separated. The aqueous was extracted with DCM (3 X 10 mL) and the combined organic layers dried with Na2SO4 and concentrated in vacuo. Purification by flash column chromatography (silica gel, 0-10% MeOH / DCM) afforded the desired product as a white solid (149 mg, 35%).

[0469] 1H NMR (400 MHz, DMSO-d6) δ 9.81 (br s, 1 H), 7.89 (dt, J = 7.6, 1.0 Hz, 2H), 7.69 (d, J = 7.5 Hz, 2H), 7.42 (td, J = 7.5, 1.2 Hz, 2H), 7.33 (td, J = 7.4, 1.2 Hz, 2H), 4.29 (d, J = 6.9 Hz, 2H), 4.21 (t, J = 6.9 Hz, 1 H), 4.03 (s, 3H), 3.88 (s, 3H), 3.26 (q, J = 6.9 Hz, 2H), 2.50 - 2.45 (m, 2H).

[0470] 13C NMR (101 MHz, DMSO-d6) δ 170.0, 158.5, 156.0, 143.9, 143.7, 140.7, 130.9, 127.6, 127.0, 125.1 , 120.1 , 96.0, 65.4, 52.4, 46.7, 40.8, 36.7, 35.3.

[0471] LRMS (ESI+, m / z) expected for Chemical Formula: C24H23BrN40s is 526.1 found 527.1 / 529.1 (M+H).

[0472] 3-(3-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-bromo-1- methyl-1H-pyrazole-5-carboxylic acid

[0473] To a stirred solution of methyl 3-(3-((((9 / 7-fluoren-9- yl)methoxy)carbonyl)amino)propanamido)-4-bromo-1-methyl-1 / 7-pyrazole-5- carboxylate (144 mg, 0.27 mmol, 1 eq) in 2.3:1 IPA / Water (16.5 mL, 0.17 M), was added anhydrous calcium chloride (1.48 g, 13.3 mmol, 48.7 eq) then lithium hydroxide (115 mg, 2.73 mmol, 10 eq). The resulting mixture was stirred at RT for 16 h. The mixture was acidified with aqueous HCI (1M) then extracted with DCM (3 X20 mL), the combined organic layers dried with Na2SC>4 and concentrated in vacuo. Purification by flash column chromatography (silica gel, 0-10% MeOH / DCM) afforded the desired product as a white solid (125 mg, 89%).

[0474] 1H NMR (400 MHz, DMSO-d6) δ 9.74 (s, 1 H), 7.89 (d, J = 7.5 Hz, 2H), 7.69 (d, J = 7.5 Hz, 2H), 7.41 (td, J = 7.5, 1.2 Hz, 2H), 7.33 (td, J = 7.4, 1.2 Hz, 2H), 4.29 (d, J = 6.8 Hz, 2H), 4.21 (t, J = 7.0 Hz, 1 H), 4.02 (s, 3H), 3.43 - 3.23 (m, 4H).

[0475] 13C NMR (101 MHz, DMSO-d6) δ 169.9, 159.6, 156.1 , 156.0, 143.9, 143.4, 140.7, 127.6, 127.1 , 125.2, 120.1 , 65.4, 46.7, 40.6, 36.6, 35.3.* LRMS (ESI+, m / z) expected for Chemical Formula: C24H23BrN40s is 512.1 found 513.1.1 / 515.1 (M+H).

[0476] *Quaternary carbon attached to the bromo is not visible.

[0477] Route to 3-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4- methyl-1-methyl-1H-pyrazole-5-carboxylic acid

[0478] Methyl 3-amino-4-methyl-1-methyl-1H-pyrazole-5-carboxylate

[0479] A mixture of K2CO3 (472 mg, 3.42 mmol, 2.0 eq), Methyl 3-amino-4-methyl-1-methyl- 1 / 7-pyrazole-5-carboxylate (400 mg, 1.71 mmol, 1.0 eq), methylboronic acid (512 mg, 8.55 mmol, 5.0 eq) and [1 ,1'-bis(diphenylphosphino)ferrocene]palladium (II) chloride (105 mg, 0.13 mmol, 7.5 mol%) was degassed (vacuum / N23 cycles) and DMF (8.55 mL, 0.2 M) was added and the resulting mixture stirred at 100 °C for 16 h. The mixture was cooled to RT, diluted with DCM, filtrated through celite then washed with 10 % aqueous LiCI. The combined organic layers were dried with Na2SO4 and concentrated in vacuo. Purification by flash column chromatography (silica gel, 0-70% EtOAc / Hexane) afforded the desired product as a beige solid (289 mg, 47%).

[0480] 1H NMR (400 MHz, CDCI3) 6 3.96 (s, 3H), 3.88 (s, 3H), 3.48 (br s, 2H), 2.10 (s, 3H).13C NMR (101 MHz, CDCI3) 6 161.3, 151.6, 130.1 , 107.5, 51.7, 39.2, 8.9.

[0481] Methyl 3-(3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-methyl- 1-methyl-1H-pyrazole-5-carboxylate Fmoc-β-Ala-OH (287 mg, 0.92 mmol, 1.2 eq), 4-dimethylaminopyridine (113 mg, 0.92 mmol, 1.2 eq) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (177 mg, 0.92 mmol, 1.2 eq) were dissolved in DCM (3.84 mL, 0.20 M) and the resulting mixture stirred at RT for 5 min. Methyl 3-amino-4-methyl-1-methyl-1H-pyrazole-5- carboxylate (130 mg, 0.77 mmol, 1 eq) was added and the resulting mixture stirred at rt for 16h. Aqueous HCI (1M) was added and the layers were separated. The aqueous was extracted with DCM (3 X 10 mL) and the combined organic layers dried with Na2SO4 and concentrated in vacuo. Purification by flash column chromatography (silica gel, 0-10% MeOH / DCM) afforded the desired product as a white solid (355 mg, 33%).

[0482] 1H NMR (400 MHz, DMSO-d6) δ 9.71 (s, 1 H), 7.92 - 7.86 (m, 2H), 7.69 (d, J = 7.4 Hz, 2H), 7.41 (td, J = 7.5, 1.1 Hz, 2H), 7.37 (br s, 1 H), 7.32 (td, J = 7.5, 1.2 Hz, 2H), 4.29 (d, J = 6.6 Hz, 2H), 4.23 - 4.18 (m, 1 H), 3.97 (s, 3H), 3.85 (s, 3H), 3.28 - 3.24 (m, 2H), 2.50 - 2.45 (m, 2H), 1.99 (s, 3H).

[0483] 13C NMR (101 MHz, DMSO-d6) δ 169.9, 160.2, 156.1 , 143.9, 143.6, 140.7, 127.6, 129.8, 127.0, 125.2, 120.1 , 116.7, 65.4, 51.9, 46.7, 36.8, 35.4, 9.5.

[0484] LRMS (ESI+, m / z) expected for Chemical Formula: C25H26N4O5 is 462.2 found 463.3 (M+H).

[0485] 3-(3-((((9H-Flu°ren-9-yl)methoxy)carbonyl)amino)propanamido)-4-methyl-1- methyl-1H-pyrazole-5-carboxylic acid

[0486] To a stirred solution of methyl 3-(3-((((9 / 7-fluoren-9- yl)methoxy)carbonyl)amino)propanamido)-4-methyl-1-methyl-1 / 7-pyrazole-5- carboxylate 110 mg, 0.24 mmol, 1 eq) in 2.3:1 IPA / Water (14.4 mL, 0.17 M), was added anhydrous calcium chloride (1.29 g, 11.6 mmol, 48.7 eq) then lithium hydroxide (99.8 mg, 2.38 mmol, 10 eq). The resulting mixture was stirred at RT for 16 h. The mixture was acidified with aqueous HCI (1M) then extracted with DCM (3 X20 mL), the combined organic layers dried with Na2SC>4 and concentrated in vacuo. Purification by flash column chromatography (silica gel, 0-10% MeOH / DCM) afforded the desired product as a white solid (68.9 mg, 65%).1H NMR (400 MHz, DMSO-d6) δ 13.39 (br s, 1 H), 9.67 (s, 1 H), 7.89 (d, J = 7.5 Hz, 2H), 7.69 (d, J = 7.5 Hz, 2H), 7.43 - 7.36 (m, 2H), 7.32 (t, J = 7.3 Hz, 2H), 4.29 (d, J = 6.6 Hz, 2H), 4.21 (t, J = 6.9 Hz, 1 H), 3.97 (s, 3H), 3.27 (q, J = 6.6 Hz, 2H), 2.52 - 2.43 (m, 3H), 1.99 (s, 3H).

[0487] 13C NMR (101 MHz, DMSO-d6) δ 169.8, 161.2, 156.1 , 143.9, 143.4, 140.7, 130.8, 127.6, 127.0, 125.2, 120.1 , 116.5, 65.4, 46.7, 36.9, 35.4, 9.5.

[0488] Route to 1-(1-benzyl-1H-imidazol-2-yl)-2,2,2-trichloroethan-1-one 1-Benzyl-1H-imidazole

[0489] To a solution of imidazole (3.5 g, 51.44 mmol, 1.0 eq) in THF (10 mL) was added sodium hydride (60% in mineral oil), (1.61 g, 66.87 mmol, 1.3 eq) portion-wise at 0 °C over a period of 45 min. Benzyl bromide (10.56 g, 7.33 mL, 61.73 mmol, 1 .2 eq) was then added to the reaction mixture in one portion at the same temperature. The reaction mixture was then stirred at room temperature for 24 hours. The resultant mixture was diluted with saturated NaHCCh <aq) solution and extracted with EtOAc, washed with brine and the organic layer dried over MgSCU, filtered and purified by flash column chromatography eluting with a gradient from 30-100% EtOAc in Petroleum Ether to afford the target compound as an off white solid (3.8g, 47%).

[0490] 1H NMR (500 MHz, DMSO-d6) δ 7.76 (d, J = 1.2 Hz, 1 H), 7.39 - 7.34 (m, 2H), 7.33 - 7.28 (m, 1 H), 7.26 (dd, J = 7.0, 1.7 Hz, 2H), 7.19 (t, J = 1.2 Hz, 1 H), 5.20 (s, 2H).

[0491] 13C NMR (126 MHz, DMSO-d6) δ 138.3, 137.9, 129.2, 129.1 , 128.2, 127.9, 120.0, 49.9. 1-(1-Benzyl-1H-imidazol-2-yl)-2,2,2-trichloroethan-1-one

[0492] To a solution of trichloroacetyl chloride (4.66 g ,2.86 mL, 25.60 mmol, 1.0eq) in DCM (25 mL), cooled to 0 °C, was added N-benzylimidazole (3.68 g / 23.28 mmol, 1.1 eq) as a solution in DCM (25 mL) via cannula and the resulting solution allowed to warm up to room temperature and stirred overnight. The reaction mixture was then cooled to 0 °C and triethylamine (2.83 g, 4.03 mL, 27.93 mmol) added and the resultant mixture allowed to warm up to room temperature and stirred for a further 5 hours, absorbed onto celite and purified by flash column chromatography eluting with a gradient from 0 to 50% DCM in petroleum ether to afford the target compound (5.01 g , 71%) as an orange oil.

[0493] 1H NMR (400 MHz, CDCI3) δ 7.41 - 7.37 (m, 3H), 7.23 - 7.20 (m, 2H), 5.64 (s, 2H).13C NMR (101 MHz, CDCI3) δ 171.7, 135.3, 134.8, 130.3, 128.6, 128.0, 127.1 , 127.1 , 94.4, 52.3.

[0494] HRMS (ESI+, m / z) expected for Chemical Formula: C12H9CI3N2O Exact Mass: 301.9780 found 302.9855 (M+H (3.82 PPM)).

[0495] Route to 2,2,2-trioMoro-1 ~(1-(syclohexy^methyO-1 H-imidazoh2-yl)ethan-1-one

[0496] 1 ~(Cyclohexylmethy1)-1H-imidazote To a 250 mL flask was added imidazole (5.0g, 73.42 mmol, 1.0 eq) and freshly ground KOH (5.94 g, 10.6 mmol, 1.5 eq) in dimethyl sulfoxide (80 mL) and stirred for 3 h. The golden solution was then cooled in an ice bath, and bromomethylcyclohexane (10.3 mL, 77.1 mmol, 1.05 eq) added. The reaction was then stirred at 0 °C for a further 4 hr before being left to warm up to room temperature overnight. The reaction was then diluted with petroleum ether (80 mL) and diethyl ether (40 mL) and washed in a separating funnel with brine (3 x 150 mL). The combined organic layers were concentrated under reduced pressure and purified by flash column chromatography a gradient from 0 to 50% DCM in petroleum ether to afford the target compound (10.2 g, 85%) as an off white solid.

[0497] 1H NMR (500 MHz, DMSO-d6) δ 7.56 (s, 1 H), 7.11 (d, J = 1.2 Hz, 1 H), 6.87 (d, J = 1.1 Hz, 1 H), 3.79 (d, J = 7.1 Hz, 2H), 1.69 - 1.58 (m, 4H), 1.53 - 1.46 (m, 2H), 1.21 - 1.07 (m, 3H), 0.90 (qd, J = 11.9, 3.5 Hz, 2H).

[0498] 2,2, 2-Trichloro~1-(14^y^H>hexylmethyl)-1 H4msdazoh2~yl)efhan~1-one)

[0499] To a solution of trichloroacetyl chloride (12.5 g ,7.72 mL, 66.95 mmol, 1.1eq) in DCM (25 mL), cooled to 0 °C, was added 1-(cyclohexylmethyl)-1H-imidazole (10.0 g / 60.86 mmol, 1.0eq) as a solution in DCM (25 mL) via cannula and the resulting solution allowed to warm up to room temperature and stirred overnight. The reaction mixture was then cooled to 0 °C and triethylamine (2.83 g, 4.03 mL, 27.93 mmol) added and the resultant mixture allowed to warm up to room temperature and stirred for a further 5 hours, absorbed onto celite and purified by flash column chromatography eluting with a gradient from 0 to 50% DCM in petroleum ether to afford the target compound (13.47 g, 65%) as a clear oil.

[0500] 1H NMR (500 MHz, DMSO-d6) δ 7.78 (d, J = 0.8 Hz, 1 H), 7.37 (d, J = 0.8 Hz, 1 H), 4.29 (d, J = 7.3 Hz, 2H), 1.73 - 1.57 (m, 5H), 1.54 - 1.48 (m, 2H), 1.16 (tt, J = 11.6, 6.2 Hz, 3H), 0.99 (qd, J = 11.8, 3.5 Hz, 2H).

[0501] 13C NMR (101 MHz, CDCI3) δ 171.7, 135.2, 129.8, 127.9, 118.5, 55.1 , 38.3, 29.9, 25.6, 25.0.

[0502] HRMS (ESI+, m / z) expected for Chemical Formula: C12H15C13N2O Exact Mass: 308.0250 found 309.0321 (M+H, (-0.5PPM)) Route to tert-butyl 3-(2-(2,2,2-trichloroacetyl)-1 / 7-imidazol-1-yl)azetidine-1- carboxylate tert-Butyl 3-(1 / 7-imidazol-1-yl)azetidine-1-carboxylate

[0503] To a solution of imidazole (1.0 g, 14.47 mmol, 1.0 eq) in THF (10 mL) was added NaH (60% in mineral oil, (0.59 g, 14.67 mmol, 1.0 eq) portion-wise at O °C over a period of 45 min. terf-butyl 3-iodoazetidine-1-carboxylate (5.0 g, 17.64 mmol, 1.2 eq) was then added to the reaction mixture in one portion at the same temperature. The reaction mixture was then stirred at room temperature for 24 hours. The resultant mixture was diluted with saturated NaHCO3 (aq) solution and extracted with ethyl acetate, washed with brine and the organic layer dried over MgSCU, filtered and purified by flash column chromatography eluting with a gradient from 30-100% ethyl acetate in petroleum ether to afford the target compound as an off white solid (2.23g, 68%). tert-Butyl 3-(2-(2,2,2-trichloroacetyl)-1H-imidazol-1-yl)azetidine-1-carboxylate To a solution of trichloroacetyl chloride (1.97 g ,1.21 mL, 10.84 mmol, 1.1eq) in DCM (25 mL), cooled to 0 °C, was added 1-(cyclohexylmethyl)- 1 W-imidazole (2.2 g / 9.85 mmol, 1.0eq) as a solution in DCM (10 mL) via cannula and the resulting solution allowed to warm up to room temperature and stirred overnight. The reaction mixture was then cooled to 0 °C and triethylamine (1.21 g, 1.7 mL, 11.92 mmol) added and the resultant mixture allowed to warm up to room temperature and stirred for a further 5 hours, absorbed onto celite and purified by flash column chromatography eluting with a gradient from 0 to 50% DCM in petroleum ether to afford the target compound (1.97 g, 49%) as a light-yellow oil.

[0504] 1H NMR (400 MHz, CDCI3) δ 7.58 (d, J = 1.1 Hz, 1 H), 7.47 (d, J = 1.1 Hz, 1 H), 5.70 (tt, J = 7.8, 5.0 Hz, 1 H), 4.58 (ddd, J = 9.8, 7.8, 0.9 Hz, 2H), 4.14 - 4.07 (m, 2H), 1.48 (s, 9H).

[0505] 13C NMR (101 MHz, CDCI3) δ 172.1 , 155.5, 135.7, 130.9, 123.1 , 116.3, 94.1 , 80.2, 47.1 , 27.8.

[0506] HRMS (ESI+, m / z) expected for Chemical Formula: C13H16C13N3O3Exact Mass: 367.0257 found 369.0326 (M+H (3.98 PPM)).

[0507] Route to 2,2,2-trichloro-1-(1-(2,2-difluoroethyl)-1H-imidazol-2-yl)ethan-1-one 1-(2,2-Difluoroethyl)-1 H -imidazole

[0508] Imidazole (0.710 g, 10.42 mmol, 1.0 eq), caesium carbonate (4.0 g, 12.50 mmol, 1.2 eq) and 1 ,1-difluoro-2-iodoethane (2g, 10.42 mmol) were added to a 20 mL microwave vial which was then sealed and flushed with argon. Acetonitrile (10 mL) was then added and the vail stirred at 70 °C for 18 hours in a sand bath. The reaction was partitioned between water and ethyl acetate and the organic layer collected. The aqueous layer was extracted with ethyl acetate (2 x 30 mL) and the combined organic layers washed with brine and dried over MgSCU, filtered and purified by flash column chromatography eluting with a gradient from 30-100% ethyl acetate in petroleum ether to afford the target compound as an off white solid (0.86g, 63%).

[0509] 1H NMR (500 MHz, DMSO-d6) δ 7.66 (s, 1 H), 7.20 (s, 1H), 6.95 (d, J= 1.1 Hz, 1H), 6.46 - 6.20 (m, 1H), 4.51 (td, J = 16.0, 3.4 Hz, 2H).

[0510] 19F NMR (471 MHz, DMSO) σ -123.16.

[0511] 2,2,2-trichloro-1-(1-(2,2-difluoroethyl)-1H-imidazol-2-yl)ethan-1-one

[0512] To a solution of trichloroacetyl chloride (1.183 g, 7.2mmol) in DCM (10 mL), cooled to 0 °C, was added 1-(2,2-difluoroethyl)-1 / 7-imidazole (0.86g g, 6.51 mmol) as a solution in DCM (25 mL) via cannula and the resulting solution allowed to warm up to room temperature and stirred overnight. The reaction mixture was then cooled to 0 °C and triethylamine (0.73 g / 7.2 mmol) added and the resultant mixture allowed to warm up to room temperature and stirred for a further 5 hours, absorbed onto celite and purified by flash column chromatography eluting with a gradient from 0 to 50% DCM in petroleum ether to afford the target compound (0.32 g 69%) as a white waxy solid.

[0513] 1H NMR (500 MHz, Chloroform-d) 5 7.40 (d, J = 1.1 Hz, 1 H), 7.28 (s, 1 H), 6.10 (tt, J = 55.3, 4.0 Hz, 1 H), 4.87 - 4.73 (m, 2H).

[0514] 19F NMR (471 MHz, Chloroform-d) 5 -122.72.

[0515] HRMS (ESI+, m / z) expected for Chemical Formula: C7H5CI3F2N2O Exact mass: 275.9436 found 276.9504 (M+H (3.55 PPM)

[0516] Route to 2,2,2-trichloro-1-(1-isopropyl-1H-imidazol-2-yl)ethan-1-one 2,2,2-trichloro-1-(1-isopropyl-1H-imidazol-2-yl)ethan-1-one To a solution of trichloroacetyl chloride (4.17 g, 22.91 mmol) in DCM (25 mL), cooled to 0 °C, was added N-isopropylimidazole (2.47 g / 22.46 mmol) as a solution in DCM (25 mL) via cannula and the resulting solution allowed to warm up to room temperature and stirred overnight. The reaction mixture was then cooled to 0 °C and triethylamine (2.32 g, 22.91 mmol) added and the resultant mixture allowed to warm up to room temperature and stirred for a further 5 hours, absorbed onto celite and purified by flash column chromatography eluting with a gradient from 0 to 50% DCM in petroleum ether to afford the target compound (3.93, 69%) as an off-white solid.

[0517] 1H NMR (400 MHz, CDCI3) δ 7.40 (q, J = 1.1 Hz, 2H), 5.43 (hept, J = 6.6 Hz, 1 H), 1.54 (d, J = 6.7 Hz, 6H).

[0518] 13C NMR (101 MHz, CDCI3) δ 171.8, 134.9, 130.4, 122.5, 94.8, 49.9, 23.1.

[0519] HRMS (ESI+, m / z) expected for Chemical Formula: C8H9C13N2O Exact Mass: 253.9780 found 254.9851 (M+H, (2.76ppm)).

[0520] Route to 1-(1-(2-((tert-butyldiphenylsilyl)oxy)ethyl)-1H-imidazol-2-yl)-2,2,2- trichloroethan-1-one 1-(2-((tert-Butyldiphenylsilyl)oxy)ethyl)-1 / 7-imidazole

[0521] To a solution of imidazole (5.0 g, 73.44 mmol, 1.0 eq) in THF (10 mL) was added NaH (60% in mineral oil, (2.94 g, 73.44 mmol, 1.0 eq) portion-wise at 0 °C over a period of 45 min. (2-chloroethoxy)(terf-butyl)diphenylsilane (28.11 g, 88.13 mmol, 1.2 eq) was then added to the reaction mixture in one portion at the same temperature. The reaction mixture was then stirred at room temperature for 24 hours. The resultant mixture was diluted with saturated NaHCO3(aq) solution and extracted with ethyl acetate, washed with brine and the organic layer dried over MgSCU, filtered and purified by flash column chromatography eluting with a gradient from 30-100% ethyl acetate in petroleum ether to afford the target compound as an off white solid (12.3g, 48%).

[0522] 1H NMR (500 MHz, DMSO-d6) δ 7.64 (s, 1 H), 7.49 - 7.44 (m, 6H), 7.40 (t, J = 7.6 Hz, 4H), 7.17 (d, J = 1.3 Hz, 1 H), 6.94 (s, 1 H), 4.15 (t, J = 5.0 Hz, 2H), 3.83 (t, J = 5.0 Hz, 2H), 0.95 (s, 9H). 1-(1-(2-((tert-Buty Id i pheny I si ly I )oxy )ethy I )-1H-imidazol-2-yl)-2,2,2-trichloroethan- 1-one

[0523] To a solution of trichloroacetyl chloride (20.83 g, 38.59 mmol) in DCM (25 mL), cooled to 0 °C, was added 1-(2-((terf-butyldiphenylsilyl)oxy)ethyl)-1 / 7-imidazole (12.3 g, 35.10 mmol) as a solution in DCM (25 mL) via cannula and the resulting solution allowed to warm up to room temperature and stirred overnight. The reaction mixture was then cooled to 0 °C and triethylamine (3.91 g, 38.59 mmol) added and the resultant mixture allowed to warm up to room temperature and stirred for a further 5 hours, absorbed onto celite and purified by flash column chromatography eluting with a gradient from 0 to 50% DCM in petroleum ether to afford the target compound (12.93 g 74%) as an off white solid.

[0524] 1H NMR (400 MHz, CDCI3) δ 7.50 - 7.48 (m, 3H), 7.44 - 7.42 (m, 2H), 7.39 - 7.34 (m, 5H), 4.62 - 4.59 (m, 2H), 4.00 - 3.96 (m, 2H), 1 .03 (s, 9H).

[0525] 3C NMR (101 MHz, CDCI3) δ 171.7, 134.8, 134.8, 134.8, 132.1 , 129.9, 129.4, 128.9, 127.4, 127.4, 127.3, 94.5, 62.2, 51.3, 26.3, 18.6.

[0526] HRMS (ESI+, m / z) expected for Chemical Formula: C23H2sCi3N2O2Si Exact Mass: 494.0751found 495.0824 (M+H, (3.86 PPM). ( / ?)-1-(((9H-fluoren-9-yl)methoxy)carbonyl)pyrrolidine-3-carboxylic acid

[0527] A solution of (R)-pyrrolidine-3-carboxylic acid (200 mg, 1.74 mmol, 1.0 eq) in 1 :1 1 ,4- dioxane / (Na2COs) aq (10 w / v%) was stirred at 0 °C for 15 min. Fmoc-succinimide (703 mg, 2.08 mmol, 1.2 eq) was then added portion-wise and the resulting mixture stirred at RT for 18 h. The reaction was then acidified with aq HCI (1 M) and extracted with ethyl acetate (3 x 20 mL). The combined organic layers were dried with Na2SO4, filtered, then concentrated in vacuo. Purification by flash column chromatography (silica gel, 0-10% MeOH / DCM) afforded the desired product as a white solid (534 mg, 91%).

[0528] 1H NMR (400 MHz, CDCI3) δ 7.77 (dt, J = 7.5, 1.0 Hz, 2H), 7.60 (dq, J = 7.5, 1.0 Hz, 2H), 7.43 - 7.37 (m, 2H), 7.33 (d, J = 1.2 Hz, 2H), 4.49 - 4.32 (m, 2H), 4.25 (t, J = 7.1 Hz, 1 H), 3.78 - 3.65 (m, 2H), 3.63 - 3.54 (m, 1 H), 3.53 - 3.43 (m, 1 H), 3.15 (dt, J = 18.7, 7.1 Hz, 1 H), 2.30 - 2.17 (m, 2H).

[0529] 13C NMR (101 MHz, CDCI3) δ 178.0, 154.9, 144.2, 141.5, 127.8, 127.2, 125.2, 120.1 , 67.6, 48.2, 47.4, 45.5, 42.27, 28.8.

[0530] (2S,4 / ?)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-1-acetylpyrrolidine-2- carboxylic acid

[0531] To a solution of (2S,4S)-4-((((9 / 7-fluoren-9-yl)methoxy)carbonyl)amino)-1-(tert- butoxycarbonyl)pyrrolidine-2-carboxylic acid (300 mg, 0.66 mmol, 1.0 eq) in DCM (5.0 mL, 0.13 M) at 0 °C was added Trifluoroacetic acid (0.50 mL, 0.66 mmol, 1.0 eq) and the resulting mixture stirred at rt for 16 h. The mixture was concentrated in vacuo and diluted again in DCM (3.31 mL, 0.2 M). The resulting mixture was cooled to 0 °C and acetic anhydride (75.2 pL, 0.80 mmol, 1.2 eq) was added dropwise. The reaction mixture was stirred at RT for 24 h. The mixture was concentrated in vacuo and purified by flash column chromatography (silica gel, 0-10% MeOH / DCM) to afford the desired product as a white solid (48.0 mg, 18%).1H NMR (400 MHz, CDCI3) δ 9.80 (br s, 1 H), 7.73 (d, J = 7.4 Hz, 2H), 7.55 (d, J = 7.5 Hz, 2H), 7.38 (t, J = 7.3 Hz, 2H), 7.29 (t, J = 7.3 Hz, 2H), 5.63 (br s, 1 H), 4.58 (br s, 1 H), 4.51 - 4.25 (m, 3H), 4.16 (br s, 1 H), 3.85 (br s, 1 H), 3.44 - 3.29 (m, 1 H), 2.47 - 2.36 (m, 1 H), 2.27 - 2.10 (m, 1 H), 2.08 (s, 3H).

[0532] 13C NMR (101 MHz, CDCI3) δ 173.3, 172.2, 156.2, 143.8, 141.5, 141.4, 127.9, 127.2, 125.1 , 120.2, 120.1 , 67.0, 58.0, 50.3, 47.2, 22.0.

[0533] LRMS (ESI+, m / z) expected for Chemical Formula: C22H22N2O5 is 394.2 found 395.2 (M+H).

[0534] HRMS (ESI): [M+H]+found 395.1604, [C22H23N2O5]+requires 395.1602.

[0535] Synthesis of polyamides

[0536] General Procedures

[0537] All reagents and solvents from commercial suppliers were used without further purification, unless otherwise stated. The monomers Fmoc-Pyrrole-OH and Fmoc- Imidazole-OH were obtained commercially from Fujifilm wako chemicals Europe GmbH and used without further purification. Solvents were of HPLC or peptide grade and used without further purification, unless otherwise stated.

[0538] Manual solid phase polyamide synthesis

[0539] Manual polyamide synthesis was carried out in a Biotage Initiator^- microwave synthesizer. Fmoc-protected resin was used (ChemMatrix®, Tentagel® S, R, or XV RAM) with DMF as a solvent and 20% v / v piperidine in DMF was used as the Fmoc deprotection solution. Synthesis was carried out in stepwise cycles of deprotection and coupling reactions, followed by cleavage from the resin and purification.

[0540] To a fritted gravity flow column equipped with a PTFE spigot was added dry resin (200 mg). The resin was swollen with DCM for 10 minutes, then drained. To the vessel was added piperidine in DMF (20% v / v) and the mixture was shaken for 3 minutes at room temperature, then drained and rinsed with DMF. The above deprotection was repeated for 10 minutes, followed by draining and rinsing with DMF. The resin was transferred to a dry 0.5-2 mL microwave vial equipped with a stirrer bar. In a separate vial was premixed the next monomer to be coupled e.g. Fmoc-Pyrrole-OH, Fmoc-lmidazole-OH, Novel monomer, etc. (4 equivalents / coupling, 0.2M, DMF or 1 :1 DM F: DM SO) DIG (4 equivalents / coupling, 0.5M, DMF), and HOBt monohydrate (4 equivalents / coupling, 0.5M, DMF). The mixture was shaken at room temperature for 5 minutes and then transferred to the resin-containing vial and heated under microwave irradiation at 75 °C for 30 minutes with slow stirring. The mixture was then transferred back to a fritted gravity flow column and subjected to the next deprotection / coupling cycle. The final imidazole coupling was instead carried out using 222-trichloro-1-methyl-1H-imidazole carboxylate (4 equivalents / coupling, 0.2M, DMF) and DIPEA (40 equivalents / coupling, 2M, DMF) via shaking at room temperature for 4-16 hours.

[0541] In some instances, novel monomers were protected with an Alloc protecting group instead of an Fmoc protecting group. In these cases, the 20% piperidine Fmoc deprotection step subsequent to the coupling of that monomer was replaced with an Alloc deprotection consisting of the following: Addition of Tetrakis(triphenylphosphine)palladium(0) (6 mol%, 0.05 M in DCM) and phenylsilane (3.75 equivalents, 0.5M in DCM) to the reaction cavity, followed by microwave irradiation at 40 °C for 5 minutes. Following this, the resin was washed with DCM and then resubjected to fresh additions of Tetrakis(triphenylphosphine)palladium(0) and phenylsilane and irradiation at 40 °C for 5 minutes.

[0542] Cleavage from the resin was carried out via shaking at room temperature in a mixture of TFA: triisopropylsilane:β-mercaptoethanol:water (92.5:2.5:2.5:2.5;v / v) for 1 hour, then washing of the resin with the same mixture. The crude solution was concentrated under a stream of compressed air then purified.

[0543] Automated solid phase polyamide synthesis

[0544] Automated polyamide synthesis was performed on a Biotage I nitiator+ Alstra automated peptide synthesis instrument. Fmoc-protected resin was used (ChemMatrix®, Tentagel® S, R, or XV RAM) with DMF as a solvent and 20% v / v piperidine in DMF was used as the Fmoc deprotection solution.

[0545] To a 10 mL microwave reactor vessel was added dry resin (200 mg) which had been swelled with DCM and drained. The appropriate amount of reagents and solvents were dispensed into the reagent tubes according to the in-built calculation software to afford the following solutions to be added automatically: Monomer e.g. Fmoc-Pyrrole-OH, Fmoc-lmidazole-OH, Novel monomer, etc. (4 equivalents / coupling, 0.2M, DMF or 1 :1 DMF:DMSO), DIC (4 equivalents / coupling, 0.5M, DMF), HOBt monohydrate (4 equivalents / coupling, 0.5M, DMF), and DIPEA (40 equivalents / coupling, 2M, DMF). The synthesis was programmed for automated stepwise deprotection and coupling reactions as follows: Fmoc deprotection of the resin (or resin-bound monomer) using 20% piperidine in DMF for 3 minutes, rinse with DMF for 30 seconds, Fmoc deprotection as above for 10 minutes, rinse with DMF for 30 seconds. Premixing of the Fmoc protected monomer with DIC and HOBt monohydrate in a separate vial followed by addition to the reaction chamber and microwave irradiation at 75 °C for 30 minutes. A pause was programmed following the penultimate deprotection step and addition of the terminal imidazole group was carried out via addition of 222-trichloro-1-methyl-1H-imidazole carboxylate and DI PEA to the reaction chamber and microwave irradiation at 75 °C for 1 hour.

[0546] In some instances, novel monomers were protected with an Alloc protecting group instead of an Fmoc protecting group. In these cases, the 20% piperidine Fmoc deprotection step subsequent to the coupling of that monomer was replaced with an Alloc deprotection consisting of the following: Addition of Tetrakis(triphenylphosphine)palladium(0) (6 mol%, 0.05 M in DCM) and phenylsilane (3.75 equivalents, 0.5M in DCM) to the reaction cavity, followed by microwave irradiation at 40 °C for 5 minutes. Following this, the resin was washed with DCM and then resubjected to fresh additions of Tetrakis(triphenylphosphine)palladium(0) and phenylsilane and irradiation at 40 °C for 5 minutes.

[0547] Cleavage from the resin was carried out via shaking at room temperature in a mixture of TFA: triisopropylsilane:β-mercaptoethanol:water (92.5:2.5:2.5:2.5;v / v) for 1 hour, then washing of the resin with the same mixture. The crude solution was concentrated under a stream of compressed air then purified.

[0548] Biotage® Selekt reverse phase purification

[0549] Purification method 1 was carried out using a Biotage® Selekt flash chromatography instrument under reverse phase conditions. Purifications were carried out on a Biotage Star C18 12g 100 A 30 pm snap cartridge. The crude mixture obtained from resin cleavage was diluted in a small volume of DMSO then injected onto the cartridge. The method employed a gradient of 15-40% 0.1% TFA spiked acetonitrile:water at a flow rate of 12mL / min over 11 column volumes with compound detection at 310 nm. The purified solution was then directly lyophilized to afford the desired compound as a solid. UltiMate™ 3000 Semi-Preparative reverse phase purification

[0550] Purification method 2 was carried out on a Thermo Scientific™ UltiMate™ 3000 SemiPreparative HPLC system using a Luna® Omega 100 A 5 pm Polar C18 column (150 x 21.2 mm). The crude mixture obtained from resin cleavage or the solid obtained from purification method 1 was diluted in a small volume of DMSO then injected onto the system rheodyne. The method employed a gradient of 15-50% 0.1 % TFA spiked acetonitrile:water at a flow rate of 17mL / min over 25 minutes with compound detection at 310 nm. The purified solution was then directly lyophilized to afford the desired compound as a solid.

[0551] Analytical HPLC methods

[0552] Analytical method 1 was carried out on a Thermo Scientific™ UltiMate™ 3000 SemiPreparative HPLC system using an AERIS™ 100 A 2.6 pm PEPTIDE XB-C18 column (250 x 4.5 mm) operated at 60 °C. The method employed a gradient of 5-95% 0.1% TFA spiked acetonitrile:water at a flow rate of 1 mL / min over 30 minutes with compound detection at 310 nm.

[0553] Analytical method 2 was carried out on a Thermo Scientific™ UltiMate™ 3000 SemiPreparative HPLC system using an AERIS™ 100 A 2.6 pm PEPTIDE XB-C18 column (250 x 4.5 mm) operated at 60 °C. The method employed a gradient of 5-50% 0.1% TFA spiked acetonitrile:water at a flow rate of 1 mL / min over 15 minutes with compound detection at 310 nm.

[0554] VI 1

[0555] ( / ?)-N-(5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)-1-methyl-4-(3-(1-methyl-4-(1-methyl-1H-imidazole-2-carboxamido)- 1H-pyrrole-2-carboxamido)propanamido)-1H-imidazole-2-carboxamide

[0556]

[0557] VI 1 was produced using fully automated microwave synthesis methods described previously using TentaGel XV RAM™ resin (200 mg, 270 pmol / g, 54.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0558] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.19 min, 94%. m / z(ESI+) 1223 ([M+H]+, 100%), 612 ([M+2H]2+, 10%)

[0559] VI 2

[0560] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0561]

[0562] VI 2 was produced using fully automated microwave synthesis methods described previously using TentaGel XV RAM™ resin (200 mg, 270 pmol / g, 54.0 pmol). The polyamide was then cleaved from the solid support using reagent K^and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0563] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.77 min, 93%. m / z(ESI+) 1222 ([M+H]+, 100%), 612 ([M+2H]2+, 11%)

[0564] VI 3 N-(5-((3-((5-((5-(((1-(2-((5-((5-((5-((3-((5-carbamoy 1-1-methyl-1H-pyrrol-3-yl)amino)- 3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol- 3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-2- oxoethyl)cyclobutyl)methyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1- methyl-1H-imidazole-2-carboxamide

[0565]

[0566] VI 3 was produced using fully automated microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (100 mg, 410 pmol / g, 41.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by preparative FPLC (Purification Method 1) and fractions lyophilised to give a yellow solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a yellow solid.

[0567] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt14.47 min, 97% m / z (ESI+) 1247 ([M+H]+, 100%), 922 (100), 624 ([M+2H]2+, 40)

[0568] VI 4 N-(5-((3-((5-((5-(((1-(2-((5-((5-((5-((3-((5-carbamoy 1-1-methyl-1H-pyrrol-3-yl)amino)- 3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol- 3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-2- oxoethyl)cyclopentyl)methyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1- methyl-1H-imidazole-2-carboxamide

[0569]

[0570] VI 4 was produced using fully automated microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (100 mg, 410 pmol / g, 41.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by preparative FPLC (Purification Method 1) and fractions lyophilised to give a yellow solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a yellow solid.

[0571] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt14.47 min, 97% m / z (ESI+) 1261 ([M+H]+, 100%), 631 ([M+2H]2+, 35)

[0572] VI 5 N-(5-((3-((5-((5-(((1-(2-((5-((5-((5-((3-((5-carbamoy 1-1-methyl-1H-pyrrol-3-yl)amino)- 3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol- 3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-2- oxoethyl)cyclohexyl)methyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1- methyl-1H-imidazole-2-carboxamide

[0573]

[0574] VI 5 was produced using fully automated microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (100 mg, 410 pmol / g, 41.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by preparative FPLC (Purification Method 1) and fractions lyophilised to give a yellow solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a yellow solid.

[0575] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt15.16 min, 99% m / z (ESI+) 1297 ([M+Na]+, 42%), 1275 ([M+H]+, 100), 638 ([M+2H]2+, 43)

[0576] VI 6

[0577] ( / ?)-N-(5-((3-((5-((5-((4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3-yl)amino)- 3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol- 3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3-hydroxy-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1 H- imidazole-2-carboxamide

[0578]

[0579] VI 6 was produced using fully automated microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (100 mg, 410 pmol / g, 41.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by preparative FPLC (Purification Method 1) and fractions lyophilised to give a yellow solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a yellow solid.

[0580] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt11.45 min, 97% m / z (ESI+) 1245 ([M+Na]+, 12%), 1223 ([M+H]+, 100), 612 ([M+2H]2+, 36)

[0581] VI 7

[0582] (S)-N-(5-((3-((5-((5-((4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3-yl)amino)- 3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol- 3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3-hydroxy-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1 H- imidazole-2-carboxamide

[0583]

[0584] VI 7 was produced using fully automated microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (100 mg, 410 pmol / g, 41.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by preparative FPLC (Purification Method 1) and fractions lyophilised to give a yellow solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a yellow solid.

[0585] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt11.48 min, 98% m / z (ESI+) 1245 ([M+Na]+, 9%), 1223 ([M+H]+, 100), 612 ([M+2H]2+, 8)

[0586] VI 8

[0587] Methyl 3-(2-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)-

[0588] 4-(1-methyl-4-(3-(1-methyl-4-(1-methyl-1H-imidazole-2-carboxamido)-1H-pyrrole- 2-carboxamido)propanamido)-1H-pyrrole-2-carboxamido)-1H-pyrrol-1- yl)propanoate

[0589]

[0590] VI 8 was produced using fully automated microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (100 mg, 410 pmol / g, 41.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by preparative FPLC (Purification Method 1) and fractions lyophilised to give a yellow solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a yellow solid.

[0591] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt11.19 min, 97% m / z (ESI+) 1316 ([M+Na]+, 11%), 1294 ([M+H]+, 100), 648 ([M+2H]2+, 90), 581 (50)

[0592] VI 9

[0593] ( / ?)-N-(5-((3-amino-4-((5-((3-((5-((5-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-2,2-dimethyl-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-3-(3-(1-methyl-4-(1- methyl-1H-imidazole-2-carboxamido)-1H-pyrrole-2-carboxamido)propanamido)-

[0594] 1 H-pyrazole-5-carboxamide

[0595]

[0596] VI 9 was produced using fully automated microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (100 mg, 410 pmol / g, 41.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by preparative FPLC (Purification Method 1) and fractions lyophilised to give a yellow solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a yellow solid.

[0597] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt11.68 min, 98% m / z (ESI+) 1273 ([M+Na]+, 7%), 1251 ([M+H]+, 100), 626 ([M+2H]2+, 67)

[0598] VI 48

[0599] VI 48 was produced using fully automated microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (100 mg, 410 pmol / g, 41.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by preparative FPLC (Purification Method 1) and fractions lyophilised to give a yellow solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a yellow solid.

[0600] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.52 min, 98% m / z (ESI+) 1245 ([M+Na]+, 7%), 1223 ([M+H]+, 100), 612 ([M+2H]2+, 38)

[0601] VI 10

[0602] (R)-3-(4-(2-amino-4-(1-methyl-4-(1-methyl-4-(3-(1-methyl-4-(1-methyl-1 H- imidazole-2-carboxamido)-1H-pyrrole-2-carboxamido)propanamido)-1H-pyrrole- 2-carboxamido)-1H-pyrrole-2-carboxamido)butanamido)-1-methyl-1H-pyrrole-2- carboxamido)-N-(5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-pyrazole-5- carboxamide

[0603] VI 10 was produced using fully automated microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (100 mg, 410 pmol / g, 41.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by preparative FPLC (Purification Method 1) and fractions lyophilised to give a yellow solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a yellow solid.

[0604] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.20 min, 98% m / z (ESI+) 1223 ([M+H]+, 18%), 926 (14), 757 (100), 612 ([M+2H]2+, 7)

[0605] VI 11 N-(5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-(1-methyl-4-(1-methyl-4-(3-(1-methyl-4- (1-methyl-1H-imidazole-2-carboxamido)-1H-pyrrole-2- carboxamido)propanamido)-1H-pyrrole-2-carboxamido)-1H-pyrrole-2- carbonyl)piperidine-4-carboxamide

[0606] VI 11 was produced using fully automated microwave synthesis methods described previously using TentaGel XV RAM™ resin (200 mg, 270 pmol / g, 54.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0607] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt12.67 min, 98%. m / z(ESI+) 1233 ([M+H]+, 100%), 936 (31), 617 ([M+2H]2+, 26)

[0608] VI 12 N-(5-((( / ?)-3-((5-((5-((( / ?)-3-am ino-4-((5-((5-((5-((( / ?)-3-((5-carbamoy 1-1-methyl-1 pyrrol-3-yl)amino)-2-hydroxy-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-4-oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)amino)-2-hydroxy-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)-1-methyl-1H-imidazole-2-carboxamide

[0609] VI 12 was produced using fully automated microwave synthesis methods described previously using TentaGel XV RAM™ resin (200 mg, 270 pmol / g, 54.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0610] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.50 min, 87%. m / z(ESI+) 1254 ([M+H]+, 100%), 628 ([M+2H]2+, 66)

[0611] VI 13 N-(5-(((S)-3-((5-((5-((( / ?)-3-amino-4-((5-((5-((5-(((S)-3-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)amino)-2-hydroxy-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-4-oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)amino)-2-hydroxy-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)-1-methyl-1H-imidazole-2-carboxamide

[0612]

[0613] VI 13 was produced using fully automated microwave synthesis methods described previously using TentaGel XV RAM™ resin (200 mg, 270 pmol / g, 54.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0614] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.26 min, 98%. m / z(ESI+) 1254 ([M+H]+, 100%), 628 ([M+2H]2+, 47)

[0615] VI 14

[0616] ( / ?)-N-(5-((3-((3-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)phenyl)amino)-3-oxopropyl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2-carboxamide

[0617]

[0618] VI 14 was produced using fully automated microwave synthesis methods described previously using TentaGel XV RAM™ resin (200 mg, 270 pmol / g, 54.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0619] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.89 min, 99%. m / z(ESI+) 1219 ([M+H]+, 100%), 610 ([M+2H]2+, 32), 581 (20).

[0620] VI 15 N-(5-((3-(((3S,5 / ?)-5-((5-((( / ?)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0621]

[0622] VI 15 was produced using fully automated microwave synthesis methods described previously using TentaGel XV RAM™ resin (150 mg, 270 pmol / g, 40.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0623] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt9.43 min, 90%. m / z (ESI+) 1211 ([M+H]+, 100%), 606 ([M+2H]2+, 25%)

[0624] HRMS (ESI+) [M+H]+found 1212.5546, [C56H7OOH N2I]+requires 1212.5558.

[0625] VI 16

[0626] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0627] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)thiophen-2-yl)amino)-3-oxopropyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2-carboxamide

[0628]

[0629] VI 16 was produced using fully automated microwave synthesis methods described previously using TentaGel R RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0630] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.19 min, 99%. m / z (ESI+) 1224 ([M+H]+, 100%), 613 ([M+2H]2+, 8%)

[0631] HRMS (ESI+) [M+H]+found 1225.4848, [C56H65N2oOnS]+requires 1225.4857.

[0632] VI 17

[0633] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0634] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-(3-methoxypropyl)-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0635]

[0636] VI 17 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0637] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.89 min, 97%. m / z (ESI+) 1279 ([M+H]+, 100%), 640 ([M+2H]2+, 11 %)

[0638] HRMS (ESI+) [M+H]+found 1280.5828, [C6OH740I2N2I]+requires 1280.5820.

[0639] VI 18 N-(5-((3-(((3R,5S)-5-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0640]

[0641] VI 18 was produced using fully automated microwave synthesis methods described previously using TentaGel R RAM™ resin (200 mg, 220 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0642] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt9.42 min, 98%. m / z (ESI+) 1211 ([M+H]+, 100%), 606 ([M+2H]2+, 12%)

[0643] HRMS (ESI+) [M+H]+found 1212.5543, [C56H7OOH N2I]+requires 1212.5558.

[0644] VI 19

[0645] (R)-N-(5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)-1-methyl-

[0646] 1 H-pyrrol-3-yl)-4-bromo-1-methyl-3-(3-(1-methyl-4-(1-methyl-1H-imidazole-2- carboxamido)-1H-pyrrole-2-carboxamido)propanamido)-1H-pyrazole-5- carboxamide

[0647]

[0648] VI 19 was produced using fully automated microwave synthesis methods described previously using TentaGel R RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0649] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.47 min, 92%. m / z (ESI+) 1300 ([M+H]+, 42%), 581 ([M+2H]2+, 100%)

[0650] HRMS (ESI+) [M+2H]2+ found 651.2275, [C56H67BrOnN22]2+ requires 651.2266.

[0651] VI 20

[0652] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0653] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-(3-cyanopropyl)-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0654]

[0655] VI 20 was produced using fully automated microwave synthesis methods described previously using TentaGel R RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0656] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.62 min, 92%. m / z (ESI+) 1276 ([M+H]+, 77%), 638 ([M+2H]2+, 100%)

[0657] HRMS (ESI+) [M+H]+found 1275.5654, [C6OH7IOH N22]+requires 1275.5667.

[0658] VI 21

[0659] Methyl (R)-3-(2-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0660] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-4-(3-(1-methyl-4-(1-methyl-1H-imidazole-2- carboxamido)-1H-pyrrole-2-carboxamido)propanamido)-1H-pyrrol-1- yl)propanoate

[0661]

[0662] VI 21 was produced using fully automated microwave synthesis methods described previously using TentaGel R RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0663] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.88 min, 68%. m / z (ESI+) 1292 ([M+H]+, 100%), 647 ([M+2H]2+, 29%)

[0664] HRMS (ESI+) [M+H]+found 1294.5569, [C6OH720I3N2I]+requires 1294.5613.

[0665] *Upon acidic analysis, part of the ester hydrolyses to the acid. Combined purity accounts for 93%.

[0666] VI 22

[0667] Methyl (R)-4-(2-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0668] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-4-(3-(1-methyl-4-(1-methyl-1H-imidazole-2- carboxamido)-1H-pyrrole-2-carboxamido)propanamido)-1H-pyrrol-1-yl)butanoate

[0669] VI 22 was produced using fully automated microwave synthesis methods described previously using TentaGel R RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0670] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt11.11 min, 52%.* m / z (ESI+) 1306 ([M+H]+, 100%), 654 ([M+2H]2+, 33%)

[0671] *Upon acidic analysis, part of the ester hydrolyses to the acid. Combined purity accounts for 94%.

[0672] VI 23

[0673] ( / ?)-4-(2-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0674] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-4-(3-(1-methyl-4-(1-methyl-1H-imidazole-2- carboxamido)-1H-pyrrole-2-carboxamido)propanamido)-1H-pyrrol-1-yl)butanoic acid

[0675]

[0676] To a solution of VI 22 (-half of the material) in MeOH (0.5 mL), was added NaOH (0.500 mL, 1.00 mmol) 2 M aq. the mixture was sonicated for 1 min to ensure the polyamide was all in solution and the resulting mixture stirred at rt for 18 h. The mixture was then acidified with TFA and directly purified by semi-preparative HPLC (Purification Method 2). The fractions were lyophilised to give a white solid.

[0677] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.22 min, 95%. m / z (ESI+) 1293 ([M+H]+, 100%), 647 ([M+2H]2+, 17%). HRMS (ESI+) [M+H]+found 1294.5546, [C6iH740i3N2i]+requires 1294.5613.

[0678] VI 24

[0679] (R)-N-(5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)-1 ,4-dimethyl-3-(3-(1-methyl-4-(1-methyl-1H-imidazole-2- carboxamido)-1H-pyrrole-2-carboxamido)propanamido)-1H-pyrazole-5- carboxamide

[0680]

[0681] VI 24 was produced using fully automated microwave synthesis methods described previously using TentaGel R RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0682] RP-HPLC (HPLC Method 2, Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.78 min, 92%. m / z (ESI+) 1236 ([M+H]+, 100%), 618 ([M+2H]2+, 26%)

[0683] HRMS (ESI+) [M+H]+found 1237.5492, [C57H69OnN22]+requires 1237.5511.

[0684] VI 25

[0685] (R)-3-(2-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0686] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-4-(3-(1-methyl-4-(1-methyl-1H-imidazole-2- carboxamido)-1H-pyrrole-2-carboxamido)propanamido)-1H-pyrrol-1-yl)propanoic acid

[0687]

[0688] To a solution of VI 21 (-half of the material) in MeOH (0.5 mL), was added NaOH (0.500 mL, 1.00 mmol) 2 M aq. the mixture was sonicated for 1 min to ensure the polyamide was all in solution and the resulting mixture stirred at rt for 18 h. The mixture was then acidified with TFA and directly purified by semi-preparative HPLC (Purification Method 2). The fractions were lyophilised to give a white solid.

[0689] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.06 min, 94%. m / z (ESI+) 1279 ([M+H]+, 100%), 640 ([M+2H]2+, 40%). HRMS (ESI+) [M+H]+found 1280.5413, [C59H7OOI3N2I]+requires 1280.54

[0690] VI 26

[0691] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0692] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-(2-(2-methoxyethoxy)ethyl)-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1 H- imidazole-2-carboxamide

[0693] VI 26 was produced using fully automated microwave synthesis methods described previously using TentaGel R RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0694] RP-HPLC (HPLC Method 2, Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt11.25 min, 96%. m / z (ESI+) 1309 ([M+H]+, 100%), 655 ([M+2H]2+, 16%)

[0695] HRMS (ESI+) [M+H]+found 1310.5907, [C6iH760i3N2i]+requires 1310.5926.

[0696] VI 27 N-(5-(((R)-3-((5-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol- 3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-2-methyl-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1 H- imidazole-2-carboxamide

[0697]

[0698] VI 27 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0699] RP-HPLC (HPLC Method 2, Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.95 min, 95%. m / z (ESI+) 1235 ([M+H]+, 100%), 618 ([M+2H]2+, 10%)

[0700] HRMS (ESI+) [M+H]+found 1236.5558, [CssHyoOnlXh? requires 1236.5558.

[0701] VI 28 N-(5-(((S)-3-((5-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol- 3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-2-methyl-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1 H- imidazole-2-carboxamide

[0702]

[0703] VI 28 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0704] RP-HPLC (HPLC Method 2, Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt11.13 min, 95%. m / z (ESI+) 1235 ([M+H]+, 100%), 618 ([M+2H]2+,28%)

[0705] HRMS (ESI+) [M+H]+found 1236.5558, [C58H70O11N21requires 1236.5558.

[0706] VI 29

[0707] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-((3-((3-

[0708] (dimethylamino)propyl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0709] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0710]

[0711] VI 29 was produced using fully automated microwave synthesis methods described previously using TentaGel XV RAM™ resin (200 mg, 160 pmol / g, 32.0 pmol). The polyamide was then cleaved from the solid support using TFA:ethanethiol:TIPS:water (92.5:2.5:2.5:2.5) and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0712] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.50 min, 96%. m / z (ESI+) 1376.4 ([M+H]+, 100%), 689.2 ([M+2H]2+, 7)

[0713] VI 30

[0714] ( / ?)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0715] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-(2-hydroxyethyl)-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0716]

[0717] VI 30 was produced using manual microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0718] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.26 min, 99%.

[0719] HRMS (ESI+) [M+H]+found 1252.5496, [C58H7OOI2N2I]+requires 1252.5507.

[0720] VI 31

[0721] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-(2-hydroxyethyl)-1H-imidazole- 2-carboxamide

[0722]

[0723] VI 31 was produced using manual microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (88.8 mg, 410 pmol / g, 35.5 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0724] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.22 min, 99%. m / z (ESI+) 1250 ([M+H]+, 100%), 626 ([M+2H]2+, 77%)

[0725] HRMS (ESI+) [M+H]+found 1252.5475, [C58H7OOI2N2I]+requires 1252.5507.

[0726] VI 32

[0727] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0728] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-benzyl-1H-imidazole-2- carboxamide

[0729]

[0730] VI 32 was produced using manual microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (88.8 mg, 410 pmol / g, 35.5 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0731] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt14.12 min, 91 %. m / z (ESI+) 1297 ([M+H]+, 60%), 649 ([M+2H]2+, 67%).

[0732] VI 33

[0733] ( / ?)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-(cyclohexylmethyl)-l H- imidazole-2-carboxamide

[0734]

[0735] VI 33 was produced using manual microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (88.8 mg, 410 pmol / g, 35.5 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0736] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt13.68 min, 91 %. m / z (ESI+) 1302 ([M+H]+, 67.7%), 652 ([M+2H]2+, 55%).

[0737] HRMS (ESI+) [M+H]+found 1304.6161 , [C63H78O11N21requires 1304.6184.

[0738] VI 34

[0739] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0740] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-(thiophen-2-yl)-1H-imidazole-2- carboxamide

[0741]

[0742] VI 34 was produced using manual microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (88.8 mg, 410 pmol / g, 35.5 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0743] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt12.68 min, 91 %. m / z (ESI+) 1289 ([M+H]+, 100%), 645 ([M+2H]2+, 38%).

[0744] HRMS (ESI+) [M+H]+found 1304.6161 , [C6OH680H N2IS]+requires 1290.5122.

[0745] VI 35

[0746] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0747] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-(thiazol-2-yl)-1H-imidazole-2- carboxamide

[0748]

[0749] VI 35 was produced using manual microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (88.8 mg, 410 pmol / g, 35.5 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0750] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt11.77 min, 92%. m / z (ESI+) 1289 ([M+H]+, 100%), 646 ([M+2H]2+, 28%).

[0751] HRMS (ESI+) [M+H]+found 1291.5065, [C59H67OnN22S]+requires 1291.5075.

[0752] VI 36

[0753] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0754] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-(azetidin-3-yl)-1H-imidazole-2- carboxamide

[0755]

[0756] VI 36 was produced using manual microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (88.8 mg, 410 pmol / g, 35.5 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0757] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt9.61 min, 94%. m / z (ESI+) 1261 ([M+H]+, 100%), 632 ([M+2H]2+, 56%).

[0758] HRMS (ESI+) [M+H]+found 1263.5653, [C59H7IOH N22]+requires 1263.5667.

[0759] VI 37

[0760] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0761] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-isopropyl-1H-imidazole-2- carboxamide

[0762]

[0763] VI 37 was produced using manual microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (88.8 mg, 410 pmol / g, 35.5 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0764] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt11.21 min, 94%. m / z (ESI+) 1248 ([M+H]+, 100%), 625 ([M+2H]2+, 26%).

[0765] HRMS (ESI+) [M+H]+found 1250.5686, [Cs^OnlXh? requires 1250.5715.

[0766] VI 38

[0767] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-

[0768] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-(2,2-difluoroethyl)-1H-imidazole- 2-carboxamide

[0769]

[0770] VI 38 was produced using manual microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (88.8 mg, 410 pmol / g, 35.5 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0771] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.78 min, 93%. m / z (ESI+) 1270 ([M+H]+, 100%), 636 ([M+2H]2+, 37%).

[0772] VI 39

[0773] ( / ?)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-ethyl-1H-imidazole-2- carboxamide

[0774]

[0775] VI 39 was produced using manual microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (88.8 mg, 410 pmol / g, 35.5 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by MPLC (Purification Method 1) and fractions lyophilised to give a white solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0776] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt9.17 min, 88%. m / z (ESI+) 1235 ([M+H]+, 100%), 618 ([M+2H]2+, 34%).

[0777] HRMS (ESI+) [M+H]+found 1236.5537, [CssHyoOnlXh? requires 1236.5558.

[0778] VI 40 VI 40 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0779] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt9.61 min, 94%. m / z (ESI+) 1211 ([M+H]+, 100%), 606 ([M+2H]2+, 26%)

[0780] HRMS (ESI+) [M+H]+found 1212.5536, [C56H7OOH N2I]+requires 1212.5558.

[0781] VI 41 N-(5-((3-(((3S,5R)-5-((5-(((R)-3-amino-4-((5-((3-((5-((5-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0782] VI 41 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid. RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt9.63 min, 94%. m / z (ESI+) 1211 ([M+H]+, 100%), 606 ([M+2H]2+, 10%)

[0783] HRMS (ESI+) [M+H]+found 1212.5538, [C56H7OOH N2I]+requires 1212.5558.

[0784] VI 42

[0785] ( / ?)-Af-(5-((3-((5-((5-((3-amino-4-((5-((3-((5-((5-((5-carbamoyl-1-methyl-1 / 7-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1 H- imidazole-2-carboxamide

[0786] VI 42 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0787] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.73 min, 95%. m / z (ESI+) 1221 ([M+H]+, 100%), 611 ([M+2H]2+, 7%)

[0788] HRMS (ESI+) [M+H]+found 1222.5382, [C57H680H N2I]+requires 1222.5402. VI 43 N-(5-((3-(((3S,5R)-5-((5-(((R)-3-amino-4-((5-(((3R,5S)-5-((5-((3-((5-carbamoyl-1- methyl-1H-pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)pyrrolidin-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0789] VI 43 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0790] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt8.12 min, 97%. m / z (ESI+) 1201 ([M+H]+, 100%), 601 ([M+2H]2+, 46%).

[0791] HRMS (ESI+) [M+H]+found 1202.5695, [C55H720H N2I]+requires 1202.5715.

[0792] VI 44 N-(5-((3-(((3R,5S)-5-((5-(((R)-3-amino-4-((5-(((3S,5R)-5-((5-((3-((5-carbamoyl-1- methyl-1H-pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)pyrrolidin-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0793]

[0794] VI 44 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0795] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt8.12 min, 97%. m / z (ESI+) 1201 ([M+H]+, 100%), 601 ([M+2H]2+, 46%).

[0796] HRMS (ESI+) [M+H]+found 1202.5691 , [C55H720H N2I]+requires 1202.5715.

[0797] VI 45 N-(5-((3-(((3S,5S)-5-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0798]

[0799] VI 45 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0800] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt9.68 min, 97%. m / z (ESI+) 1210 ([M+H]+, 100%), 606 ([M+2H]2+, 76%). HRMS (ESI+) [M+H]+found 1202.5535, [C56H7OOH N2I]+requires 1212.5558.

[0801] VI 46 N-(5-((3-(((3S,5S)-5-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0802]

[0803] VI 46 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0804] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt9.48 min, 92%. m / z (ESI+) 1210 ([M+H]+, 100%), 606 ([M+2H]2+,55%). HRMS (ESI+) [M+H]+found 1212.5529, [C56H7OOH N2I]+requires 1212.5558.

[0805] VI 47 N-(5-((3-((R)-3-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol- 3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-1-yl)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0806]

[0807] VI 47 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAM™ resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0808] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt10.06 min, 92%. m / z (ESI+) 1196 ([M+H]+, 100%), 598 ([M+2H]2+, 7%). HRMS (ESI+) [M+H]+found 1197.5427, [C56H690nN2o]+requires 1197.5449.

[0809] VI 48

[0810] (R)-N-(5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)-1-methyl-

[0811] 1 H-pyrrol-3-yl)-1-methyl-3-(3-(1-methyl-4-(1-methyl-1H-imidazole-2-carboxamido)- 1H-pyrrole-2-carboxamido)propanamido)-1H-pyrazole-5-carboxamide

[0812]

[0813] VI 48 was produced using fully automated microwave synthesis methods described previously using ChemMatrix® H-Rink Amide resin (100 mg, 410 pmol / g, 41.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified to -80% by preparative FPLC (Purification Method 1) and fractions lyophilised to give a yellow solid. An analytical sample was then purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a yellow solid.

[0814] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 10.52 min, 98% m / z (ESI+) 1245 ([M+Na]+, 7%), 1223 ([M+H]+, 100), 612 ([M+2H]2+, 38)

[0815] VI 49

[0816] N-(5-((3-(((1 R,3S)-3-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)cyclopentyl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide VI 49 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0817] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 10.32 min, 94%. m / z (ESI+) 1210 ([M+H]+, 100%).

[0818] HRMS (ESI+) [M+H]+found 1211.5600, [C57H7IOH N2O]+requires 1211.5606.

[0819] VI 50

[0820] N-(5-((3-(((1 S,3R)-3-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)cyclopentyl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0821] VI 50 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0822] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 10.37 min, 99%. m / z (ESI+) 1210 ([M+H]+, 100%), 605 ([M+2H]2+, 6%).

[0823] HRMS (ESI+) [M+H]+found 1211.5575, [C57H7IOH N2O]+requires 1211.5606.

[0824] VI 51

[0825] N-(5-((3-(((3R,5R)-5-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0826] VI 51 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0827] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 9.44 min, 93%. m / z (ESI+) 1211 ([M+H]+, 100%).

[0828] HRMS (ESI+) [M+H]+found 1212.5529, [C56H7OOH N2I]+requires 1212.5558.

[0829] VI 52 N-(5-((3-(((3R,5R)-5-((5-(((R)-3-amino-4-((5-((3-((5-((5-((5-carbamoyl-1-methyl-1 H- pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0830] VI 52 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0831] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 9.58 min, 95%. m / z (ESI+) 1210 ([M+H]+, 100%).

[0832] HRMS (ESI+) [M+H]+found 1212.5534, [C56H7OOH N2I]+requires 1212.5558.

[0833] VI 53

[0834] N-(5-((3-((S)-3-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1 H-pyrrol- 3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-1-yl)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0835]

[0836] VI 53 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0837] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 10.10 min, 98%. m / z (ESI+) 1196 ([M+Hl+, 100%). HRMS (ESI+) [M+H]+found 1197.5421 , [C56H690nN2o]+requires 1197.5449.

[0838] VI 54

[0839] N-(5-((3-(((3R,5R)-5-((5-(((R)-3-amino-4-((5-((R)-3-((5-((3-((5-carbamoyl-1-methyl-

[0840] 1 H-pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)pyrrolidine-1-carbonyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide VI 54 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0841] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 8.58 min, 98%. m / z (ESI+) 1185 ([M+H]+, 100%), 593 ([M+2H]2+, 18%).

[0842] HRMS (ESI+) [M+H]+found 1187.5575, [C55H7IOH N2O]+requires 1187.5611.

[0843] VI 55

[0844] N-(5-((3-(((3R,5R)-5-((5-(((R)-3-amino-4-((5-((S)-3-((5-((3-((5-carbamoyl-1-methyl- 1 H-pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)pyrrolidine-1-carbonyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0845] VI 55 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0846] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 8.58 min, 96%. m / z (ESI+) 1185 ([M+H]+, 100%), 593 ([M+2H]2+, 32%).

[0847] HRMS (ESI+) [M+H]+found 1187.5580, [C55H7IOH N2O]+requires 1187.5611.

[0848] VI 56

[0849] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)- 1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-(2, 5, 8,11 ,14-pentaoxahexadecan-16-y l)-1H- pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H- imidazole-2-carboxamide

[0850] VI 56 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (400 mg, 222 pmol / g, 88.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0851] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 11.30 min, 97%. m / z (ESI+) 1439 ([M+H]+, 53%), 720 ([M+2H]2+, 100%).

[0852] HRMS (ESI+) [M+H]+found 1442.6681 , [CeyHssOieN^P requires 1442.6726. VI 57

[0853] N-(5-((3-(((1 R,3S)-3-((5-(((R)-3-amino-4-((5-((3-((5-((5-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)cyclopentyl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0854] VI 57 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMerand the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0855] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 10.52 min, 91%. m / z (ESI+) 1210 ([M+H]+, 100%), 605 ([M+2H]2+, 26%).

[0856] HRMS (ESI+) [M+2H]2+found 606.2834, [CsyFWu^o]2* requires 606.2839.

[0857] VI 58

[0858] N-(5-((3-(((1 S,3R)-3-((5-(((R)-3-amino-4-((5-((3-((5-((5-((5-carbamoyl-1-methyl-1H- pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)cyclopentyl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2- carboxamide

[0859]

[0860] VI 58 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0861] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 10.53 min, 95%. m / z (ESI+) 1210 ([M+HJ+, 100%), 605 ([M+2H]2+, 40%). HRMS (ESI+) [M+H]+ found 1211.5575, [C57H71O11 N20]+ requires 1211.5611.

[0862] VI 59

[0863] N-(5-((3-(((3R,5S)-1-acetyl-5-((5-(((R)-3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1- methyl-1H-pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-4-oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H- imidazole-2-carboxamide

[0864]

[0865] VI 59 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0866] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 10.07 min, 90%. m / z (ESI+) 1252 ([M+H]+, 100%), 627 ([M+2H]2+, 17%). HRMS (ESI+) [M+H]+found 1254.5789, [C58H72OI2N2I]+requires 1254.5664.

[0867] VI 60

[0868] (R)-N-(5-((3-((5-((5-((3-amino-4-((5-((5-((5-((3-((5-carbamoyl-1-methyl-1H-pyrrol-3- yl)amino)-3-oxopropyl)carbamoyl)-1-(2-(2-methoxyethoxy)ethyl)-1H-pyrrol-3- yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-(2-(2-methoxyethoxy)ethyl)- 1 H-pyrrol-3-yl)amino)-4-oxobutyl)carbamoyl)-1-(2-(2-methoxyethoxy)ethyl)-1H- pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1- (2-(2-methoxyethoxy)ethyl)-1H-pyrrol-3-yl)-1-methyl-1H-imidazole-2-carboxamide

[0869]

[0870] VI 60 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (400 mg, 222 pmol / g, 88.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0871] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 12.12 min, 98%. m / z (ESI+) 1572 ([M+H]+, 100%), 787 ([M+2H]2+, 27%). HRMS (ESI+) [M+2H]2+found 787.8837, [C73H101 O19N21F requires 787.8786.

[0872] VI 61

[0873] N-(5-((3-(((3S,5S)-5-((5-(((R)-3-amino-4-((5-(((R)-3-((5-((5-((5-carbamoyl-1-methyl-

[0874] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H- pyrrol-3-yl)amino)-2-hydroxy-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-4-oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H- imidazole-2-carboxamide

[0875]

[0876] VI 61 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0877] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 9.42 min, 92%. m / z (ESI+) 1226 ([M+H]+, 100%), 613 ([M+2H]2+, 32%). HRMS (ESI+) [M+H]+found 1228.5641 , [C56H7OOI2N2I]+requires 1228.5507.

[0878] VI 62

[0879] N-(5-((3-(((3S,5S)-5-((5-(((R)-3-amino-4-((5-(((S)-3-((5-((5-((5-carbamoyl-1-methyl-

[0880] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H- pyrrol-3-yl)amino)-2-hydroxy-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-4-oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H- imidazole-2-carboxamide

[0881]

[0882] VI 62 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0883] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 9.41 min, 90%. m / z (ESI+) 1227 ([M+H]+, 58%), 614 ([M+2H]2+, 100%). HRMS (ESI+) [M+H]+found 1228.5579, [C56H7OOI2N2I]+requires 1228.5507.

[0884] VI 63

[0885] N-(5-((3-(((3S,5S)-5-((5-(((R)-3-amino-4-((5-(((S)-3-((5-((5-((5-carbamoyl-1-methyl-

[0886] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H- pyrrol-3-yl)amino)-2-methyl-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-4-oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H- imidazole-2-carboxamide

[0887]

[0888] VI 63 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0889] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 9.94 min, 91%. m / z (ESI+) 1224 ([M+H]+, 100%), 613 ([M+2H]2+, 10%). HRMS (ESI+) [M+H]+found 1226.5785, [C57H720H N2I]+requires 1226.5715.

[0890] VI 64

[0891] N-(5-((3-(((3S,5S)-5-((5-(((R)-3-amino-4-((5-(((R)-3-((5-((5-((5-carbamoyl-1-methyl-

[0892] 1 H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H- pyrrol-3-yl)amino)-2-methyl-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3- yl)amino)-4-oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3- yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-1-methyl-1H- imidazole-2-carboxamide

[0893]

[0894] VI 64 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (200 mg, 222 pmol / g, 44.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMerand the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0895] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 9.95 min, 95%. m / z (ESI+) 1224 ([M+H]+, 100%), 613 ([M+2H]2+, 20%). HRMS (ESI+) [M+H]+found 1226.5800, [C57H720H N2I]+requires 1226.5715.

[0896] VI 65

[0897] 5-(3-(6-((2-((5-((3-(((3S,5S)-5-((5-(((R)-3-amino-4-((5-((3-((5-((5-((5-carbamoyl-1- methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl- 1 H-pyrrol-3-yl)amino)-3-oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)amino)-4- oxobutyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)pyrrolidin-3-yl)amino)-3- oxopropyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)carbamoyl)-1-methyl-1H-imidazol- 4-yl)amino)-6-oxohexyl)thioureido)-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid

[0898]

[0899] VI 65 was produced using fully automated microwave synthesis methods described previously using TentaGel S RAMTM resin (300 mg, 222 pmol / g, 66.0 pmol). The polyamide was then cleaved from the solid support using reagent KpMer and the volatiles removed under a stream of air. The crude material was purified by semi-preparative HPLC (Purification Method 2) and fractions lyophilised to give a white solid.

[0900] RP-HPLC (HPLC Method 1 , Aeris Peptide XB-C18, 2.6 pm, 100 A, 250 x 4.6 mm, 310 nm) Rt 12.74 min, 92%. m / z (ESI+) 1727 ([M+H]+, 18%), 864 ([M+2H]2+, 100%).

[0901] HRMS (ESI+) [M+2H]2+found 865.3484, [C83H94Oi7N24S]2+requires 865.3469.

[0902] RESULTS

[0903] Experimental conditions for Dynamic Biosensors switchSENSE® methods

[0904] Experimental overview: The reported data was generated via real-time detection of a change in fluorescent signal of a dye connected to a gold surface on a prefabricated chip via a DNA linker. The chip is coated with single stranded DNA via a thiol linkage and can be modularly functionalized via double stranded DNA hybridization with nanolevers that introduce the required functionality for the assay. The heliX instrument and associated chips and buffers were commercially supplied by dynamic biosensors. Required DNA was either supplied by dynamic biosensors or contracted for synthesis by an external CRO. Upon chip functionalization, the dye is proximal to a double stranded DNA binding site (herein referred to as the ligand) to which DNA-binding compounds (herein referred to as analytes) can bind and influence fluorescence via proximity quenching. Pre-hybridization of DNA probes (ligand strand):

[0905] DNA oligos were produced containing - at the 3'-end - a generic 48 mer switchSENSE immobilization sequence (5'-ATC AGT ACT TGT CAA CAC GAG CAG CCC GTA TAT TCT CCT ACA GCA CTA-3', SEQ ID NO: 1), complementary to the adaptor strand which is hybridised to the thiol-tethered DNA on the switchSENSE chip - and at the 5'-end - the corresponding target sequence as an overhang connected to the generic sequence by a tri-thymine spacer and terminated by a GCG stabilisation triplet (EG19 and EG20). Short oligos complementary to the individual overhang sequences were also synthesized (EG19C and EG20C). All DNA oligos used for switchSENSE experiments were synthesized by Ella Biotech GmbH (Furstenfeldbruck, Germany). To yield DNA probes with double-stranded overhang regions the ligand strand (EG19 or EG20) and the corresponding overhang complement strand (EG19C or EG20C) were mixed at an equal concentration (500 nM) in PE40 buffer (10 mM Na2HPO4 / NaH2PO4, 40 mM NaCI, 0.05 % Tween20, 50 pM EDTA, 50 pM EGTA, pH 7.4) and shaken at 600 rpm at 25 °C for 20 minutes. This was then combined with and equal volume of the dye-tagged adaptor strand (AS1-Ra, Dynamic Biosensors GmbH, Germany, 400 nM) and hybridized under the same conditions. For immobilization of DNA probes on switchSENSE chips, the prehybridized DNA probes were hybridized to the surface grafted complement of the anchor sequence using an inbuilt routine of the heliX instrument.

[0906] Oligo sequence definitions:

[0907] EG19+EG19C: GC-containing the WT-dsDNA binding site for GS compounds attached to switchSENSE immobilization sequence.

[0908] EG 19: 5'-GCG CTC AGA AGT TTT TTT ATC AGT ACT TGT CAA CAC GAG CAG CCC GTA TAT TCT CCT ACA GCA CTA-3'. SEQ ID NO: 2

[0909] EG19C: 5'-AAA ACT TCT GAG CGC-3', SEQ ID NO: 3

[0910] EG20+EG20C: AT-containing the SNP-dsDNA binding site for GS compounds attached to switchSENSE immobilization sequence.

[0911] EG20: 5'-GCG CTC AGA AAT TTT TTT ATC AGT ACT TGT CAA CAC GAG CAG CCC GTA TAT TCT CCT ACA GCA CTA-3', SEQ ID NO: 4

[0912] EG20C: 5'-AAA ATT TCT GAG CGC-3', SEQ ID NO: 5 Analyte (GS compound) preparation:

[0913] A small arbitrary amount of the GS analyte was taken up in DMSO, diluted first into 1 :1 DMSO:water, then into water, and the concentration was determined by NanoDrop ND- 1000 UV-Vis Spectrophotometer. The DMSO stock was then diluted with the experiment running buffer to the appropriate concentration for the experiment.

[0914] Kinetics determination:

[0915] Binding experiments of analytes to ligands were carried out using a heliX switchSENSE platform on a gold sensor chip (Dynamic Biosensors GmbH, Germany). PE140 buffer (10 mM Na2HPO4 / NaH2PO4, pH 7.4, 140 mM NaCI, 0.05 % Tween20, 50 pM EDTA, 50 pM EGTA, pH 7.4) served as running buffer for the interaction experiments and for hybridization and on-chip immobilization of the DNA nanolevers. The association and the dissociation kinetics of the GS compounds to the DNA probes was measured under a flow rate of 50 pl / min analyte solution and buffer, respectively. Binding traces, corresponding to the absolute fluorescence intensity readout, were recorded using the instrument’s static measurement mode. In this measurement mode, a constant mild negative potential is applied to the gold chip surface, which results in an upright orientation of the DNA nano-levers. Every set of association and dissociation was referenced to a buffer blank injection recorded prior to the analyte injection. For data analysis, the kinetic data sets were processed using heliOS software (Dynamic Biosensors GmbH, Germany).

[0916] The kinetic rate constants were determined in the heliOS software with the following equations:

[0917] Association Formula

[0918] Dissociation Equilibrium Formula

[0919] Parameters y sensor signal tonassociation start

[0920] A amplitude konassociation rate constant c concentration toffdissociation start koffdissociation rate constant yO baseline

[0921] KDdissociation equilibrium constant

[0922] Table 1 : Table showing kinetic measurements for association and dissociation rate constants, and association and dissociation equilibrium constants for compounds of the invention, measured against the EG20+EG20C SNP DNA sequence (on-target) via the Dynamic Biosensors switchSENSE® assay delineated above. Data obtained in duplicate with ± values representing standard deviation from the mean. Entries lacking ± values were obtained in singlicate at concentrations of 500pM, 1 nM, and 2nM. .

[0923] Table 2: Table showing kinetic measurements for association and dissociation rate constants, and association and dissociation equilibrium constants for GS compounds, measured against the EG19+EG19C WT-DNA sequence (off-target) via the Dynamic Biosensors switchSENSE® assay delineated above. Data obtained in duplicate with ± values representing standard deviation from the mean. Entries lacking ± values were obtained in singlicate at concentrations of 500pM, 1nM, and 2nM.

[0924] Table 3: Table comparing the dissociation equilibrium constants of GS compounds against both EG20+EG20C (SNP) (on-target) and EG19+EG19C (WT) (off-target) sequences. The selectivity coefficient for each GS compound is a ratio of the KD against EG19+EG19C divided by the KD against EG20+EG20C. Data obtained in duplicate with ± values representing standard deviation from the mean. Entries lacking ± values were obtained in singlicate at concentrations of 500pM, 1nM, and 2nM.

[0925] SUMMARY

[0926] These assays represent the first use of the DBS switchSENSE® method via real-time detection to quantify the different binding kinetics of hairpin PAs against two ds-DNA sequences (i.e. one containing the WT sequence flanking position 9477 from the COXI 11 gene (EG19) and the single SNP where the GC base pair has been replaced with AT (EG20)).

[0927] Two PAs comprising traditional monomer units with either a Py / lm at the B2 / B5position of formula (I) (VI 1) selective for the WT GC base pair or a Py / Py at B2 / B5position of formula (I) (VI 2) selective for the SNP AT base pair were compared with other compounds of formula (I) to assess how modified monomer units introduced at various positions within the compound changed the binding affinity for the target EG 20 SNP sequence compared with the ‘off-target’ EG 19 WT sequence. These data demonstrate that the compounds of the invention display differing degrees of selectivity for the GC- containing dsDNA sequence EG19+EG19C, and the AT-containing dsDNA sequence EG20+EG20C, summarised in Table 3.

[0928] The data for control polyamide VI 2 demonstrates selectivity for the SNP sequence in EG 20 over the WT in EG 19 as expected and follows Dervan’s pairing rules previously defined for hairpin polyamides, where the Py / Py pairing at B2 / B5prefers binding with the SNP AT base pair over the WT GC base pair-containing sequence. For control polyamide VI 2, the Im / Py pairing demonstrates selectivity for the WT sequence, again as predicted by Dervan’s pairing rules.

[0929] VI 11 had a greater affinity for the target SNP EG20 sequence. Replacement of the D group amino moiety with a hydroxyl group (VI 7) affords similar on-target affinity and selectivity to the controls. Similarly, modifications to the Py N-Me substituent at B2over the SNP site also demonstrated comparable selectivity over the WT sequence compared with the control VI 2 polyamide (see VI 20 and VI 30). This indicates that Py N-groups introduced to modify polyamide physicochemical properties do not negatively impact Dervan’s pairing rules and retain an affinity equivalent to the control polyamide (which contains a Py N-Me motif). However, the Py N-group modification exemplified by VI 56 with a PEG motif not only retains affinity for the SN P sequence, but also notably improves selectivity over the WT compared with the control VI 2 (21.4-fold more selective versus 7.1 for control). This observation has significant implications for hairpin polyamides as a class, where PEG groups introduced into multiple Py units within a PA sequence can improve overall water solubility without impairing binding affinity or selectivity.

[0930] N-substitution at A generally had minimal impact on binding kinetics (see VI 31 , VI 38, and VI 39). Interestingly, the azetidine moiety of VI 36 afforded a 6-fold increase in affinity for both DNA sequences, leaving selectivity unaffected.

[0931] The most profound changes to binding affinity and selectivity for the SNP sequence were exhibited by replacing the Py unit at B2with a pyrrolidine monomer (novel for a hairpin polyamide). VI 15, VI 18, VI 46, and VI 51 contain the four possible chiral permutations of a pyrrolidine motif at B2. These compounds have varying degrees of improved on- target affinity, and a trend for decreased off-target affinity, with the trans-pyrrolidine displaying an 18.5-fold selectivity, compared to the 7.1-fold selectivity for the control VI 2.

[0932] Deletion of the pyrrolidine hydrogen bond donor was explored via acetylation, wherein VI 59 exhibited similar on-target affinity and improved selectivity compared to the comparable stereoisomer VI 18.

[0933] To further investigate the role of compound flexibility in the vicinity of the SNP-binding monomer, a switch in sequence was implemented between the β-ala unit at C2and the Py unit at B5, which according to Dervan’s rules should have no impact on the targeted DNA sequence. A control compound was synthesised for this sequence (VI 42) which exhibited a 3-fold increase in affinity for both sequences, but as expected, no effect on selectivity. Contrary to this, when this sequence modification was employed with a pyrrolidine unit at B2, a drastic improvement in selectivity was observed, owing to increases in on-target affinity concomitant to decreases in off-target selectivity (Comparing VI 15 to VI 40, VI 18 to VI 41 , VI 51 to VI 52, and VI 46 to VI 45). This series afforded our most potent and selective compound to date, the C2-B5switched S,S-c / s- pyrrolidine VI 45, with a 46.0 pM affinity and ~90-fold selectivity.

[0934] Functional transcription assay

[0935] To support the DNA-binding data collected by DBS, a bespoke fluorescent assay was used to measure the inhibition of polymerase-mediated transcription in vitro. The bespoke assay used a modified format of the assay previously reported by Hoefer et al. in J. Am. Chem. Soc. (2013), 135, 13692-13694. The probe used for this consists of an aptamer-coding DNA which upon transcription and dye binding produces fluorescence and a DNA target region of interest (ROI) for polyamide binding. Polyamides binding to the DNA in a way that inhibits transcription of the DNA construct results in loss of fluorescence. To dissect polyamide specificity, we routinely used two ROI constructs in tandem, one containing the wild-type sequence used in the DBS experiments (vida supra) and one containing the SNP.

[0936] Methods

[0937] To assess DNA binding of polyamides, we used an in-house transcription inhibition assay. A green fluorescent protein (GFP) mimicking RNA aptamer / dye couple was used to measure inhibition of DNA-dependent RNA polymerase (here T3) activity upon polyamide binding to a target DNA sequence. Assay principle and design was based on the publication by Hoefer et al., 2013, supra. The assay was optimised for assessment of polyamide binding with minimal background and high throughput capacity.

[0938] Polyamides were tested at 1 and 0.1 pM in a high throughput screen against DNA constructs with a ROI containing the embedded SNP EG20 sequence used in the DBS experiments (termed L below) or an off-target ROI representing the embedded WT EG 19 sequence (termed C below). Chemicals were purchased from Merck or Fisher unless otherwise stated. Custom DNA oligomers were purchased from Merck (desalt and HPLC purification). DNA was rehydrated in IDT duplexing buffer (Integrated DNA technologies, 30 mM HEPES, pH 7.5; 100 mM potassium acetate) to a stock concentration of 40 pM. DNA-dependent RNA polymerase and rNTPs were purchased from New England Biolabs.

[0939] Fluorescence was read in a Hidex sense multiplate reader at ex / em 472(+ / -30 nm) / 535(+ / -20 nm), 10 flashes, low lamp power, top read.

[0940] Results were background corrected and expressed as % relative to DNA without polyamide. A heparin curve was used as an internal inhibition control. Results were plotted and analysed using GraphPad Prism (version 9.5.1).

[0941] DNA target sequences used to evaluate effects on transcription are shown in Table 4:

[0942] After the initial high throughout screening (HTS), polyamides that showed preferential binding of the target over the non-target ROI, were tested across a concentration range from 3 nM to 10 pM in half log units to obtain IC50S for binding to on- and off-target ROIs L and C (Table 5). Most promising polyamides and new derivatives based on these were further tested across a concentration range as before, against DNA constructs containing no ROI plus a linker (noROI+GT). For all experiments, results were background corrected (DNA construct without broccoli aptamer) and expressed as % of aptamer without polyamide. A heparin curve was used as an internal inhibition control for each individual experiment to confirm validity. Experiments were conducted with three independent repeats and all experimental instances were found to be valid as judged from the heparin and background control. Results were plotted and analysed using GraphPad Prism (version 9.5.1).

[0943] DNA sequences used are shown in Table 4.

[0944] Table 4: DNA sequences of aptamer constructs used in the hybrid broccoli transcription

[0945] Results

[0946] Table 5: IC50 values in nM for polyamides tested across a concentration range. Values were calculated from graphs plotted with n=3±SEM (standard error of the mean) in GraphPad Prism 9. Heparin served as an assay control.

[0947] Hairpin polyamides were tested for sequence selective inhibition of polymerase activity. For high throughput screening, two concentrations were chosen at 1 and 0.1 pM polyamide. Experiments were conducted with three independent repeats and all experimental instances were found to be valid as judged from the heparin and background control. All polyamide results are displayed in Fig. 1 ; a collection of representative polyamides is shown in Fig. 2 to highlight the effect of different monomeric variations on pyrrole (Py), imidazole (Im), pyrazole (Pz) and β-alanine (P-ala) within the polyamide structure on inhibition. The aim was to establish whether the DNA binding kinetics exhibited for the equivalent WT and SNP sequences in the DBS binding assay (vida supra), when embedded in the template sequence, translated into a functional output by demonstrating inhibition of transcription by RNA polymerase. This is the first report on whether hairpin polyamide binding affinity to DNA sequences containing a single SNP reflects a functional downstream output (gene transcription) in a similar, selective manner.

[0948] Across all samples, the 1 pM concentrations showed the most distinct changes (Fig. 1). Representative polyamides illustrating key findings are shown in Fig. 2. Surprisingly, VI 1 and VI 2, designed using Dervan’s pairing rules to target the WT ‘C’ sequence or the SNP ‘L’ sequence, respectively, exhibited no difference in functional output: transcription of both sequences was inhibited equally and effectively at 1 pM. This contrasts with VI 1 and VI 2 having preferential binding affinity for the WT or SNP sequence, respectively (as assessed using the DBS method (Table 3)). Similarly, polyamides containing Py units with N-substituents at the B2position of formula (I) did not demonstrate particularly selective transcription inhibition. These data suggest for the first time that, whilst polyamides designed around Dervan’s pairing rules do exhibit binding selectivity at the biophysical level, this does not translate into an equivalent selective functional output when the sequences being transcribed differ only by a single SNP. This observation has significant implications for polyamides being designed and deployed for selective pharmacological use.

[0949] Remarkably, by contrast, when a pyrrolidine unit was incorporated at the B2position of formula (I) and paired with a Py at the B5position of formula (I) (VI 15 and VI 18), not only was selective binding demonstrated at the association level when assessed by DBS (Table 3), the equivalent selectivity was also translated into the functional output: both polyamides selectively inhibited transcription of the SNP sequence over the WT at 1 pM concentration, with a 4.6- and 6-fold change for VI 15 and VI 18, respectively and on- target IC50S of 232 and 139 nM, while the IC50 for the noROI+GT control was 1858 and 2353 nM, respectively. Furthermore, when Py at the B5position was replaced with a β- ala unit and paired with a conventional Py at the B2position (VI 42), transcription of both sequences was inhibited equally, contravening Dervan’s pairing rules, with IC50S of 40 nM for L and C and an IC50 of 110 nM for the noROI+GT control (Table 3). However, when the pyrrolidine monomers were positioned at B2and paired with β-ala at B5(e.g. VI 40), selective transcription inhibition of the SNP sequence was seen even at the lower concentration of 0.1 pM, with IC50S of 50 nM for on-target (L) and 385 nM for off-target (C) DNAs, which is a 7.7-fold preferential binding for the on-target sequence. The IC50 for the noROI+GT control was 2710 nM. An equivalent output was also demonstrated when a c / s-pyrrolidine unit at B2was paired with a β-ala at B5(VI 45), resulting in an unprecedented 8-fold selective binding of the target sequence (IC50 of 72 nM) over the off-target sequence (573 nM) with an IC50 of 2537 for the noROI+GT. Both of these polyamides show only a minor loss of affinity compared to the 40 nM for VI 42 and a significant increase of on-target binding compared to the noROI+GT control. Interestingly, pairing a trans-pyrrolidine at B2with another trans-pyrrolidine at B5(VI 43 and VI 44), significantly impaired the ability of the polyamide to inhibit transcription of both sequences, even at 1 pM. Taken together, our results show that very subtle changes in output can be achieved by deploying different pairing arrangements of these saturated monomers either with themselves or with conventional Py / lm / Pz / β-ala PA monomer units.

[0950] Finally, a β-proline unit at the NH-B2position of formula (I) (linked via the tertiary cyclic amide), when paired with Py at B5and exemplified by VI 47, can also discriminate between the SNP and WT sequence at the transcriptional level. β-Prolines (pyrrolidine- 3-carboxylates) or their derivatives have not previously been incorporated into hairpin polyamides.

[0951] Skin Cell Assay

[0952] Total ATP availability in a human is limited to that which can be derived from inhaled O2. To a degree, the human body can modulate ATP delivery to certain cell types / locations by reducing ATP availability at others. One consequence of temporary ATP deficiency is the acne prodrome which, without treatment, progresses to the visible marker of bacterial skin infection and pustulation. This can be alleviated by topical treatment of the acne prone skin with agents that, in combination, can alleviate or stimulate processes so that ATP synthesis is not only stimulated but “clogged” mitochondria are destroyed and disposed of so that mitochondrial renewal can occur (see US 2021 / 0361591 A1).

[0953] T o assess the efficacy of combination treatment with the polyamides of the invention and with agents that can stimulate mitochondrial renewal, the assay described below is used.

[0954] Cells are cryopreserved in liquid nitrogen. Cells are seeded in plates for differentiation with linoleic acid to increase lipid production. Optimal cell densities are identified using a luminescence assay, e.g. Cell-titer Gio™ (Promega), to evaluate ATP production, multiplexed with an Alamar Blue™ assay to evaluate mitochondrial activity. This multiplexed assay is used to screen the polyamides of the invention with agents that can stimulate mitochondrial renewal. These include a mitophagy modulator, such as urolithin A, an antioxidant, such as thiotaine (L-(+)-Ergothioneine), and a cofactor such as niacinamide. The amounts of each for use in the assay are indicated below.

Claims

CLAIMS:

1. A compound of formula (I):wherein:A is a 5-membered heterocycle optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylCs- ecycloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl; each B1to B7is independently a 5- or 6-membered heterocycle, pentanone, benzene, 5- or 6-membered aliphatic carbocycle, or ethan-1 ,2-di-yl, wherein: the 5- or 6- membered heterocycle, pentanone, benzene, and 5- or 6-membered aliphatic carbocycle are optionally substituted with any one or more selected from C1-4alkyl, C1-ealkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl-(5-membered heterocyclyl), C1-4alkyl(C1-4alkoxy)pC1-4alkoxy, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), carboxyl, CO2(C1-4alkyl), CO(C1-4alkyl), C1-4alkylCO2(C1-4alkyl), oxo, C2-6alkynyl, C^ehaloalkynyl, amino, hydroxy, C1-6alkyldiC1-4alkylamino, C1-4alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC3-6azacycloalkyl, C1-4alkylmorpholinyl, C1-4alkylCs- ehaloazacycloalkyl, phenyl, 5-membered heterocyclyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2; and the ethan-1 , 2-di-yl is optionally substituted with any one or more selected from hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy, wherein the C1-4alkyl substituent may combine with the -NH- group adjacent to the ethan-1 , 2-di-yl to form a C3-5azacycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; each C1and C2is independently selected from a 5- or 6-membered N-heterocycle and ethan-1 , 2-di-yl, wherein the 5- or 6-membered N-heterocycle is optionally substituted with any one or more selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, halo, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl-(5-memberedheterocyclyl), C1-4alkyl(C1-4alkoxy)pC1-4alkoxy, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), carboxyl, CO2(C1-4alkyl), oxo, C2-6alkynyl, C2-6haloalkynyl, amino, hydroxy, C1-4alkyldiC1-4alkylamino, Ci.6alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC1-4alkoxyC1-4alkoxy, C1-4alkylC3-6azacycloalkyl, C1-4alkylmorpholinyl, C1-4alkylC3-6haloazacycloalkyl, phenyl, 5- membered heterocyclyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2; and the ethan-1 ,2-di- yl is optionally substituted with any one or more selected from hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy; p is 0 to 5;D is selected from C3-6azacycloalkyl and aminopropyl, wherein the azacycloalkyl is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy and the aminopropyl is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy, wherein two C1-4alkyl substituents may combine to form a C2-6cycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; andR is selected from H, C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl)2, C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl)H and C1-6alkylC(O)NHC1-6alkylNH2.

2. The compound of claim 1 , wherein A is an optionally substituted N-heteroaryl comprising 1 or 2 nitrogen atoms.

3. The compound of claim 1 or claim 2, wherein A is an optionally substituted pyrrole or imidazole.

4. The compound of any one of claims 1 to 3, wherein A is of formula (Ila) or (lib):wherein:R1and R2are each independently selected from C1-4alkyl, C1-4alkylol, C1-ehaloalkyl, C1-4alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl; andR3and R4are each independently selected from hydrogen, halo, C1-4alkyl and C1-ehaloalkyl.

5. The compound of claim 4, wherein:R1is selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-2alkylphenyl, C1-2alkylC5-6cycloalkyl, C2-6azacycloalkyl, thiophenyl and thiazolyl;R2is C1-4alkyl; andR3and R4are each independently selected from H and halo.

6. The compound of claim 4, wherein A is of formula (Ila) and R1is selected from C2- ealkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylC3-6cycloalkyl, C3-6azacycloalkyl, and 5-membered heterocyclyl.

7. The compound of any one preceding claim, wherein each of B1to B7is independently selected from an optionally substituted pyrrole, pyrazole, furazan, thiophene, pyrrolidine, tetra hydrofuran, imidazolidine, piperidine, and oxazolidine; an optionally substituted pentanone; an optionally substituted benzene; an optionally substituted cyclopentane; and an optionally substituted ethan-1 ,2-di-yl.

8. The compound of any one preceding claim, wherein each of B1to B7is independently selected from formulae (Illa) to (Ilin):wherein:* labels the wavy line that bissects the bond to carbonyl;R5and R7are each independently selected from H, C1-4alkyl, C1-4alkylol, C1-ehaloalkyl, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, Ci.6alkylC1-4alkoxy, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), Ci.6alkyldiC1-4alkylamino, C1-4alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC1-4alkoxyC1-4alkoxy, C1-4alkylC3-6azacycloalkyl, C1-4alkylC1-3heteroaryl, and C1-4alkylC3-6haloazacycloalkyl;R6is selected from C1-4alkyl, C1-4haloalkyl and halo;R8is selected from H, C1-4alkyl, C1-4haloalkyl, halo, C2-6alkynyl, C2-6haloalkynyl, amino and phenyl; each X is independently selected from NR9, O and C(O); each R9is independently selected from H, C1-4alkyl, C(O)C1-4alkyl, SO(C1-4alkyl), SO2(C1-4alkyl) and SO2NH2; each R10is independently selected from C1-4alkyl, C1-4haloalkyl, halo, hydroxy, amino and C1-4alkoxy; each R11and R12is independently selected from H, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, NH2, phenyl and C1-4alkoxy, wherein the C1-4alkyl substituent may combine with the -NH- group adjacent to the ethan-1 ,2-di-yl to form a C3-5azacycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; n is 0 to 2; n1 is 0 or 1 ; and n2 is 0 to 3.

9. The compound of claim 8, wherein:R5is selected from C1-4alkyl, C1-4alkylol, C1-4haloalkyl, C1-4alkylphenyl, C1-4alkylpyridyl, C1-4alkylC3-6cycloalkyl, C1-4alkyl(C1-4alkoxy)i-5, C1-4alkylcarboxyl, C1-6alkylCO2(C1-4alkyl), C1-4alkyldiC1-4alkylamino, C1-4alkylC1-4alkylamino, C1-4alkylcyano, C1-4alkylC3-6azacycloalkyl, C1-4alkylC1-3heteroaryl, and C1-4alkylC3-6haloazacycloalkyl;R6and R7are each independently C1-4alkyl;R8is selected from H, C1-4alkyl, C1-4haloalkyl, halo, C2-6alkynyl, C2-6haloalkynyl, amino and phenyl;R11is H or C1-4alkyl, wherein the C1-4alkyl substituent may combine with the -NH- group adjacent to the ethan-1 ,2-di-yl to form a C3-5azacycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; andR12is selected from H, hydroxy, C1-4alkyl, NH2, and phenyl.

10. The compound of claim 8 or claim 9, wherein B1, B3, B4and B7are of formula (Illa), optionally wherein R5of B4and B7are each independently selected from C1-4alkyl, such as methyl.11 . The compound of any one preceding claim, wherein B3is of formula (Illa) and R5of B3is selected from C1-4alkylol, C1-4alkyl(C1-4alkoxy)i-5, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), and C1-4alkylcyano, such as C1-4alkylCO2(C1-4alkyl).

12. The compound of claim 10, wherein R5of B3is selected from C1-4alkyl, C1-4alkylCO2(C1-4alkyl), and C1-4alkylcarboxyl.

13. The compound of any one preceding claim, wherein B2is selected from: formula (Illa), wherein R5is selected from C1-4alkylol, C1-4alkyl(C1-4alkoxy)i-5, C1-4alkylcarboxyl, C1-4alkylCO2(C1-4alkyl), and C1-4alkylcyano; formula (lllc), wherein R7is C1-4alkyl and R8is selected from C1-4alkyl, C1-4haloalkyl and halo; formula (Hid); formula (Illg), wherein X is NR9, wherein R9is selected from H and C(O)C1-4alkyl and each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo; formula (IIIm), wherein one of R11and R12is ethyl and combines with the -NH- group adjacent to the ethan-1 ,2-di-yl to form pyrrolidinyl that is optionally substituted with any one or more selected from C1-4alkyl, halo and C1-4haloalkyl; and formula (Ilin), wherein each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo.

14. The compound of any one of claims 8 to 13, wherein B5is of formula (Illa), (I I Ic), (Illg) or (IIIm), optionally wherein:R5of B5is selected from C1-4alkyl, C1-4alkylcarboxyl and C1-4alkylCs-6azacycloalkyl;R7and R8of B5are each independently selected from H and C1-4alkyl;X is NH and each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo andR11and R12of B5are each independently selected from H and C1-4alkyl.

15. The compound of any one preceding claim, wherein B5is: of formula (Illg), wherein X is NH and each R10is independently selected from C1-4alkyl, C1-4haloalkyl and halo; or of formula (IIIm), wherein R11is C1-4alkyl and R12is H or C1-4alkyl.

16. The compound of any one of claims 8 to 15, wherein B6is selected from formula (Illa), (Illg) and (IIIm), optionally wherein:R5of B6is C1-4alkyl, such as methyl, or C1-4alkyl(C1-4alkoxy)i-5;R11of B6is H; andR12of B6is selected from H, hydroxy, C1-4alkyl, NH2, and phenyl.

17. The compound of any one preceding claim, wherein the 5- or 6-membered N- heterocycle of C1and C2is pyrrolidine.

18. The compound of any one preceding claim, wherein each C1and C2is independently selected from formulae (IVa) to (IVc):wherein:* labels the wavy line that dissects the bond to carbonyl;R5is C1-4alkyl, such as methyl; andeach R11and R12is independently selected from H, hydroxy, C1-4alkyl, NH2, phenyl, halo, C1-4haloalkyl and C1-4alkoxy.

19. The compound of claim 18, wherein C1and C2are independently selected from formulae (IVa) and (IVc) and optionally wherein R11of formula (IVc) is H.

20. The compound of any one of claims 1 to 19, wherein C1is of formula (IVc), R11is H and R12is independently selected from C1-4alkyl, C1-4haloalkyl and C1-4alkoxy.

21. The compound of any one preceding claim, wherein D is selected from C3-6azacycloalkyl and aminopropyl, wherein the azacycloalkyl is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy and the aminopropyl is substituted with two C1-4alkyl substituents combined to form a C4- ycycloalkane that is optionally substituted with any one or more selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy.

22. The compound of any one of claims 1 to 20, wherein D is selected from formulae (Va) to (Vc):wherein:R13is selected from H, NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy;R14is selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy;R15is selected from NH2, hydroxy, halo, C1-4alkyl, C1-4haloalkyl, and C1-4alkoxy; m is 1 ; and o is 0 to 4.

23. The compound of claim 22, wherein D is of formula (Va).

24. The compound of any one preceding claim, wherein R is selected from H, and C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl)2, such as H.

25. A compound of formula (1), (2), (3), or (4):wherein:A, B1to B7, C1, C2, D and R are as defined in any one preceding claim; R’ is selected from C1-6alkylC(O)NHC1-6alkylN(C1-6alkyl) and C1-6alkylC(O)NHC1-6alkylNH; R2’ is selected from NHC(O)C1-6alkylNH, C(O)NHC1-6alkylNH, NHC(O)Ci.6alkylC(O) and C(O)NHC1-6alkylC(O); andM is a mitochondrial delivery agent or a detectable tag.

26. A mitochondria-targeting capsule comprising a compound of any one preceding claim.

27. A pharmaceutical composition comprising a compound of any one of claims 1 to 25 or a mitochondria-targeting capsule of claim 26 and a pharmaceutically acceptable excipient.

28. The compound of any one of claims 1 to 25, mitochondria-targeting capsule of claim 26, or pharmaceutical composition of claim 27 for use in medicine, as a medicament or in a method of treatment.

29. The compound of any one of claims 1 to 25, mitochondria-targeting capsule of claim 26, or pharmaceutical composition of claim 27 for use in a method of treating or preventing mitochondrial dysfunction or a disease or condition associated therewith.

30. A method of treating or preventing mitochondrial dysfunction or a disease or condition associated therewith in a subject comprising administering a therapeuticallyeffective amount of a compound of any one of claims 1 to 25, mitochondria-targeting capsule of claim 26, or pharmaceutical composition of claim 27 to the subject.

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