Novel compounds and therapeutic uses thereof

Abstract

The invention relates to novel compounds with the ability to link an immune response to a pathogen, to the use of said compounds in a disease or disorder mediated and / or caused by an infective agent, to compositions containing said compounds, processes for their preparation and to novel intermediates used in said process.

Description

[0001] CEN-C-P3772PCT NOVEL COMPOUNDS AND THERAPEUTIC USES THEREOFFIELD OF THE INVENTION The invention relates to novel compounds with the ability to link an immune response to apathogen, to the use of said compounds in a disease or disorder mediated and / or caused byan infective agent, to compositions containing said compounds, processes for theirpreparation and to novel intermediates used in said process.BACKGROUND OF THE INVENTION There is a need to find novel ways to recruit an individual’s immune system to fight disease. The human immune system continually surveys the body seeking foreign signals to identify potentially harmful pathogens or mutated human cells (that could become a cause ofcancerous growth) and target them for elimination. Natural antibodies exist that can berecruited to said pathogens or mutated human cells to drive the immune system to eliminatethe threat. The invention details the use of a novel set of compounds that are designed toattract these natural antibodies in such a way as to be able to maximise the efficacy of immune recruitment while minimising potential side effects. There is an urgent need to identify novel ways of treating bacterial, viral and fungal infections. Drug resistance is becoming a major global health threat. For example, more than 2 million people in the US were infected with bacteria resistant to at least one class of antibiotics (Centers for Disease Control and Prevention, 2013). Overall, the identification of new antibiotics targeting resistant strains of gram-negative organisms has been particularly difficult, in part due to the complex and evolving strategy these bacteria use to prevent antibiotic action (e.g., production of antibiotic inactivating enzymes, ability to transfer of resistance between strains, efflux pumps to prevent intracellular action) coupled with theirnaturally impermeable cell membranes that make it hard to identify drugs that penetrate intothe cell and inhibit key targets. Further, many strains utilize multiple resistance mechanisms making it difficult for a single antibiotic to overcome. An innovative approach to the treatment of infectious disease was disclosed in WO 01 / 45734 which describes a set of novel immunity linkers. Examples of said linker moietiesinclude compounds or agents which are recognised by the immune system of said individualas foreign and which would therefore trigger an immune response. One such example is acarbohydrate molecule capable of binding to a human anti-alpha-Gal antibody e.g.galactosyl-alpha-1,3-galactosyl-beta-1,4-N-acetylglucosamine) which results in redirection ofthe natural human serum antibody anti-alpha-gal. The resultant effect of said immunity linker CEN-C-P3772PCT molecule is that the immune response of the individual is diverted from the pre-existing immune response of said individual towards the target, i.e. the pathogen.More recently, a further example of a carbohydrate molecule capable of binding to humanserum antibodies has been described. For example, anti-Rhamnose (i.e. L-Rhamnose) hasbeen shown to result in target cell destruction (Kiessling et al (2014) ChemBioChem 15(10),1393-1398; US 2014 / 0112975; Li et al (2016) ACS Chem. Biology 11(5), 1205-1209).Furthermore, complexes comprising anti-Rhamnose antibodies have been shown to magnify antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependentcytotoxicity (CDC) which therefore confirm utility in the anti-cancer field (Zhou et al (2022)Journal of Medicinal Chemistry 65, 323-332).There is therefore a need for alternative compounds for the treatment of a disease ordisorder mediated and / or caused by an infective agent. SUMMARY OF THE INVENTION According to a first aspect of the invention, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof: wherein: F is selected from: rhamnose; or a carbohydrate molecule capable of binding to a human anti-alpha-galactosyl antibody; S2is selected from: -(CH2)2-O-(CH2)2-NH-CO-CH2-, -(CH2)2-O-(CH2)2-NH-CO-(CH2CH2O)4-(CH2)2-NH-CO-CH2-, –(CH2)3-NH-CO-CH2-, -(CH2CH2O)2-(CH2)2-NH-CO-CH2- or –(CH2)3-CO-NH-(CH2)2-NH-CO-CH2-; Y2is -O-; R1represents: C1-6alkyl, haloC1-6alkyl or heterocyclyl; Y1is -CONH-; S1is -(CH2CH2O)8-(CH2)2-; X1is -C(=O)-;L represents a cationic anti-microbial peptide selected from a moiety of formula (L): CEN-C-P3772PCT wherein X1 represents the point of attachment of L to X1; Z1 represents C1-10 alkyl or benzyl optionally substituted by one or more halogen or haloC1-6 alkyl groups; Z2 represents -CH2-CH2-NH2, -CH2-NH2 or -CH2-OH; Z3 represents benzyl or -CH2-CH(CH3)2;Z4 represents -CH2-CH(CH3)2, -CH(CH3)-CH2CH3, -CH(OH)-CH3 or -CH2CH3; andZ5 represents -CH(OH)-CH3 or -CH2-CH(CH3)2.DETAILED DESCRIPTION OF THE INVENTIONWO 2020 / 201743 describes a conjugate which comprises a cationic anti-microbial peptide(that is shown to specifically bind to bacteria) and a carbohydrate molecule capable of bindingto a human anti-alpha-galactosyl antibody (e.g. galactosyl-alpha-1,3-galactosyl-beta-1,4-N-acetylglucosamine, alpha1-3 galactobiose, alpha1-3-beta1-4-galactotriose orgalilipentasaccharide) connected via a linker. Suitable examples of cationic peptides includepolymyxin B (or polymyxin nonapeptide, colistin or a derivative thereof). This family of cationicpeptides bind to lipid A on the bacterial cell surface and, when conjugated to alpha-gal linkers,will present alpha-gal, resulting in anti-alpha-gal antibody recruitment and cell killing.Resistance rates are likely to be low as lipid A is important in the survival of gram-negative bacteria. In fact, even polymyxin-resistant strains retain binding sites for cationic peptides andas such the peptide-alpha gal conjugate.The novel mechanisms described in WO 2020 / 201743 are particularly attractive due to notbeing impacted by antibiotic resistance mechanisms and provide effectiveness against multi- CEN-C-P3772PCTdrug resistant strains. Furthermore, the combination of the broad spectrum bacterial bindingcapability of a cationic peptide with the unique ability to specifically recruit naturally occurringanti-alpha-gal antibodies to the bacterial surface, and re-direct these antibodies to promotecomplement activation, phagocytosis and killing is very attractive. Therefore, the compoundsdescribed in WO 2020 / 201743 claimed to have the potential to provide a novel therapy forbacterial infections with broad-spectrum activity. However, antimicrobial peptides (such as polymyxin and derivatives thereof) are oftenassociated with having a liability for renal toxicity (Nation et al (2019) Antibiotics 8, 24).Surprisingly, the inventors of the present invention have discovered that an alkyl containing substituent on the phenyl ring (in place of the additional phenyl ring) provides unexpected advantages regarding reduction of cytotoxicity, in particular renal cytotoxicity. Data ispresented herein in Tables 2 to 5 which demonstrates that the alkyl substituent containingcompounds of the present invention retained the antibacterial activity (see Table 5) andantibody recruitment activity (see Tables 2 and 3) of the resultant complex but surprisinglydemonstrated significantly reduced cytotoxicity against the renal cell line hRPTEC whencompared with two representative examples from WO 2020 / 201743 (see Table 4).Furthermore, data is presented herein in Tables 6 and 7 which demonstrates the in vivoeffectiveness of selected compounds of the invention.The term ‘alkyl’ as used herein as a group or part of a group refers to a straight-chain (i.e. unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is completely saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is completely saturated or that contains one or more units of unsaturation, but which is not aromatic that has a single point of attachment to the rest of the molecule. Unless otherwise specified, alkyl groups contain 1 to 20 alkyl carbon atoms, such as 1 to 10 aliphatic carbon atoms, in particular 1 to 8 aliphatic carbon atoms, more particularly 1 to 6 alkyl carbon atoms, especially 1 to 4 alkyl carbon atoms, more especially 1 to 3 alkyl carbon atoms. Non-limiting examples of alkyl groups include, but are not limited to, linear or branched, and substituted or unsubstituted alkyl. Alkyl groups may be substituted or unsubstituted. Alkyl groups may be straight-chain or branched. The term ‘halo’ or ‘halogen’ as used herein refers to fluorine, chlorine, bromine or iodine. CEN-C-P3772PCTThe term ‘haloalkyl’ as used herein refers to a linear or branched alkyl, as the case may be,which is substituted with one or more halogen atoms. Non-limiting examples of haloalkyl groups include -CHF2, -CH2F, -CF3, -CF2-, and perhaloalkyls, such as -CF2CF3. The term ‘benzyl’ as used herein refers to a -CH2-phenyl moiety. The term ‘heterocyclyl’ as used herein refers to a monocyclic or bicyclic non-aromatic, partially saturated or fully saturated ring system containing for example 3 to 12 ring members. Each ring may contain up to five heteroatoms typically selected from nitrogen, sulfur and oxygen.Particular examples of ‘heterocyclyl’ include morpholine, piperidine (e.g. piperidin-1-yl,piperidin-2-yl, piperidin-3-yl and piperidin-4-yl), piperidinone, pyrrolidine (e.g. pyrrolidin-1-yl,pyrrolidin-2-yl and pyrrolidin-3-yl), pyrrolidone, azetidine, pyran (2H-pyran or 4H-pyran), dihydrothiophene, oxetanyl, dihydropyran, dihydrofuran, dihydrothiazole, tetrahydrofuran,tetrahydrothiophene, dioxane, tetrahydropyran (e.g. tetrahydropyran-4-yl), imidazoline,imidazolidinone, oxazoline, thiazoline, pyrazolin-2-yl, pyrazolidine, piperazinone and piperazine. In one embodiment, F represents rhamnose. The term ‘rhamnose’ as used herein refers to a compound represented by the following structure (A): wherein S2 refers to the point of attachment to the S2 group.Rhamnose (Rha, Rham) is a naturally occurring deoxy sugar and is known chemically as(2R,3R,4R,5R,6S)-6-methyloxane-2,3,4,5-tetrol (also known as isodulcit, α-L-Rhamnose, L-Rhamnose, L-Mannomethylose, α-L-Rha, α-L-Rhamnoside, α-L-Mannomethylose, 6-Deoxy-L-mannose, Rhamnopyranose and Rhamnopyranoside). It can be classified as either amethyl-pentose or a 6-deoxy-hexose. Rhamnose occurs in nature in its L-form as L- rhamnose (6-deoxy-L-mannose). It will be appreciated that minor modifications to the rhamnose moiety of the compound of the present invention while retaining the antibody recruitment properties of rhamnose are CEN-C-P3772PCT within the scope of the invention, for example, it could be envisaged that the D-Rhamnose analogue or an S-linked rhamnose analogue (i.e. a thioglycoside) may be utilised. In an alternative embodiment, F represents a carbohydrate molecule capable of binding to a human anti-alpha-galactosyl antibody.The term “carbohydrate molecule capable of binding to a human anti-alpha-galactosylantibody” as used herein refers to sugar (i.e. carbohydrate) moieties capable of binding to animmune response component (i.e. an anti-alpha-galactosyl antibody) of said human andconsequently eliciting an immune response in a human. Examples of such carbohydratemolecules include alpha-galactosyl compounds and modified derivatives thereof. Further examples of suitable carbohydrate molecules include the alpha-gal epitopes listed in US 2012 / 0003251 as being suitable for use in the selective targeting and killing of tumour cells. In one embodiment, F is selected from galactosyl-alpha-1,3-galactosyl-beta-1,4-N- acetylglucosamine, alpha1-3 galactobiose, alpha1-3-beta1-4-galactotriose or galilipentasaccharide. In one particular embodiment, F has a structure as shown in one of the following formulae: wherein S2refers to the point of attachment to the S2group. In one particular embodiment, F has a structure as shown in the following formula (B): wherein S2 refers to the point of attachment to the S2 group. CEN-C-P3772PCT In a further embodiment, F represents a compound of formula (A): wherein S2refers to the point of attachment to the S2group, or a compound of formula (B): wherein S2refers to the point of attachment to the S2group. In one embodiment, S2 is selected from: -(CH2)2-O-(CH2)2-NH-CO-CH2-, -(CH2)2-O-(CH2)2-NH-CO-(CH2CH2O)4-(CH2)2-NH-CO-CH2- or –(CH2)3-NH-CO-CH2-.In one further embodiment, R1 represents C1-6 alkyl. In a yet further embodiment, R1 represents C4 alkyl. In a still yet further embodiment, R1 represents tert butyl. In one alternative embodiment, R1 represents haloC1-6 alkyl. In a further alternative embodiment, R1 represents trifluoromethyl. In one alternative embodiment, R1 represents heterocyclyl. In a further alternative embodiment, R1 represents a 6 membered heterocyclyl ring. In a yet further alternative embodiment, R1 represents a 6 membered heterocyclyl ring containing two heteroatoms selected from nitrogen and oxygen. In a still yet further alternative embodiment, R1 represents morpholinyl. In a still yet further alternative embodiment, R1 represents N-linked morpholinyl. In one embodiment, R1 represents tert butyl, trifluoromethyl or morpholinyl (such as N-linked morpholinyl). In one embodiment, Z1 represents C1-10 alkyl (such as octyl). In an alternative embodiment, Z1represents benzyl optionally substituted by one or more halogen or haloC1-6alkyl groups. In a further embodiment, Z1represents benzyl optionally CEN-C-P3772PCT substituted by one or more halogen (such as fluorine or chlorine) or haloC1-6alkyl (such astrifluoromethyl) groups. In a further embodiment, Z1 represents 4-flurobenzyl, 4-chlorobenzylor 4-trifluoromethylbenzyl.In a yet further embodiment, Z1 represents octyl, 4-fluorobenzyl, 4-chlorobenzyl or 4-trifluoromethylbenzyl. In one embodiment, Z2represents -CH2-NH2.In one embodiment, Z3represents benzyl. In one embodiment, Z4represents -CH2-CH(CH3)2. In one embodiment, Z5represents -CH(OH)-CH3. According to a further aspect of the invention, there is provided a compound of formula (I)aor a pharmaceutically acceptable salt thereof: (I)awherein: F is selected from a compound of formula (A): wherein S2 refers to the point of attachment to the S2 group; or a compound of formula (B): CEN-C-P3772PCT wherein S2refers to the point of attachment to the S2group; S2is selected from: -(CH2)2-O-(CH2)2-NH-CO-CH2-, -(CH2)2-O-(CH2)2-NH-CO-(CH2CH2O)4- (CH2)2-NH-CO-CH2-, or –(CH2)3-NH-CO-CH2-; Y2is -O-; R1represents: tert butyl, trifluoromethyl or morpholinyl; Y1is -CONH-; S1is -(CH2CH2O)8-(CH2)2-; X1is -C(=O)-;L represents a cationic anti-microbial peptide selected from a moiety of formula (L): wherein X1represents the point of attachment of L to X1; Z1represents C1-10alkyl or benzyl optionally substituted by one or more halogen or haloC1-6alkyl groups; Z2represents -CH2-NH2; Z3represents benzyl; Z4represents -CH2-CH(CH3)2; and Z5 represents -CH(OH)-CH3. In one embodiment, the invention provides a compound of formula (I) which comprises acompound of Examples 1-7 or a pharmaceutically acceptable salt thereof. In oneembodiment, the invention provides a compound of formula (I) which is the free base or thetrifluoroacetate or acetate salt of a compound of Examples 1-7. In a further embodiment, theinvention provides a compound of formula (I) which is the free base or the acetate salt of acompound of Examples 1-7. In a yet further embodiment, the invention provides a compoundof formula (I) which is the free base of a compound of Examples 1-7. In an alternative CEN-C-P3772PCT embodiment, the invention provides a compound of formula (I) which is the acetate salt of a compound of Examples 1-7. A reference to a compound of formula (I) and sub-groups thereof also includes ionic forms, salts, solvates, isomers (including geometric and stereochemical isomers), tautomers, N- oxides, esters, isotopes and protected forms thereof, for example, as discussed below; preferably, the salts or tautomers or isomers or N-oxides or solvates thereof; and more preferably, the salts or tautomers or N-oxides or solvates thereof, even more preferably the salts or tautomers or solvates thereof. Hereinafter, compounds and their ionic forms, salts, solvates, isomers (including geometric and stereochemical isomers), tautomers, N-oxides, esters, isotopes and protected forms thereof as defined in any aspect of the invention (except intermediate compounds in chemical processes) are referred to as "compounds of the invention". Compounds of formula (I) can exist in the form of salts, for example acid addition salts or, in certain cases salts of organic and inorganic bases such as carboxylate, sulfonate and phosphate salts. All such salts are within the scope of this invention, and references to compounds of formula (I) include the salt forms of the compounds. The salts of the present invention can be synthesized from the parent compound that contains a basic moiety by conventional chemical methods such as methods described in Pharmaceutical Salts: Properties, Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002. Generally, such salts can be prepared by reacting the base forms of these compounds with the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are used. Alternatively, such salts may be prepared by utilizing ion exchange resin methodology.Acid addition salts (mono- or di-salts) may be formed with a wide variety of acids, bothinorganic and organic. Examples of acid addition salts include mono- or di-salts formed withan acid selected from the group consisting of acetic, 2,2-dichloroacetic, adipic, alginic, ascorbic (e.g. L-ascorbic), L-aspartic, benzenesulfonic, benzoic, 4-acetamidobenzoic, butanoic, (+) camphoric, camphor-sulfonic, (+)-(1S)-camphor-10-sulfonic, capric, caproic, caprylic, cinnamic, citric, cyclamic, dodecylsulfuric, ethane-1,2-disulfonic, ethanesulfonic, 2- hydroxyethanesulfonic, formic, fumaric, galactaric, gentisic, glucoheptonic, D-gluconic, glucuronic (e.g. D-glucuronic), glutamic (e.g. L-glutamic), α-oxoglutaric, glycolic, hippuric, CEN-C-P3772PCT hydrohalic acids (e.g. hydrobromic, hydrochloric, hydriodic), isethionic, lactic (e.g. (+)-L- lactic, (±)-DL-lactic), lactobionic, maleic, malic, (-)-L-malic, malonic, (±)-DL-mandelic, methanesulfonic, naphthalene-2-sulfonic, naphthalene-1,5-disulfonic, 1-hydroxy-2-naphthoic, nicotinic, nitric, oleic, orotic, oxalic, palmitic, pamoic, phosphoric, propionic, pyruvic, L- pyroglutamic, salicylic, 4-amino-salicylic, sebacic, stearic, succinic, sulfuric, tannic, (+)-L- tartaric, thiocyanic, p-toluenesulfonic, undecylenic and valeric acids, as well as acylated amino acids and cation exchange resins.One particular group of salts consists of salts formed from acetic, hydrochloric, hydriodic,phosphoric, nitric, sulfuric, citric, lactic, succinic, maleic, malic, isethionic, fumaric, benzenesulfonic, toluenesulfonic, methanesulfonic (mesylate), ethanesulfonic, naphthalenesulfonic, valeric, acetic, propanoic, butanoic, malonic, glucuronic and lactobionicacids. One particular salt is the trifluoroacetate or acetate salt, more particularly the acetatesalt. Where the compounds of formula (I) contain an amine function, these may form quaternary ammonium salts, for example by reaction with an alkylating agent according to methods well known to the skilled person. Such quaternary ammonium compounds are within the scope of formula (I).The compounds of the invention may exist as mono- or di-salts depending upon the pKa ofthe acid from which the salt is formed. The salt forms of the compounds of the invention are typically pharmaceutically acceptable salts, and examples of pharmaceutically acceptable salts are discussed in Berge et al.,1977, "Pharmaceutically Acceptable Salts," J. Pharm. Sci., Vol. 66, pp.1-19. However, saltsthat are not pharmaceutically acceptable may also be prepared as intermediate forms which may then be converted into pharmaceutically acceptable salts. Such non-pharmaceutically acceptable salts forms, which may be useful, for example, in the purification or separation of the compounds of the invention, also form part of the invention. Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as “solvates”. For example, a complex with water is known as a “hydrate”. Pharmaceutically acceptable solvates of the compound of the invention are within the scope of the invention. CEN-C-P3772PCT Compounds of formula (I) containing an amine function may also form N-oxides. A reference herein to a compound of formula (I) that contains an amine function also includes the N- oxide. Where a compound contains several amine functions, one or more than one nitrogen atom may be oxidised to form an N-oxide. Particular examples of N-oxides are the N-oxides of a tertiary amine or a nitrogen atom of a nitrogen-containing heterocycle. N-Oxides can be formed by treatment of the corresponding amine with an oxidizing agent such as hydrogen peroxide or a per-acid (e.g. a peroxycarboxylic acid), see for example Advanced Organic Chemistry, by Jerry March, 4thEdition, Wiley Interscience, pages. More particularly, N-oxides can be made by the procedure of L. W. Deady (Syn. Comm.1977, 7, 509-514) in which the amine compound is reacted with m-chloroperoxybenzoic acid (mCPBA), for example, in an inert solvent such as dichloromethane. It will be appreciated by those skilled in the art that certain protected derivatives of compounds of formula (I), which may be made prior to a final deprotection stage, may not possess pharmacological activity as such, but may, in certain instances, be administered orally or parenterally and thereafter metabolised in the body to form compounds of the invention which are pharmacologically active. Such derivatives may therefore be described as “prodrugs”. All such prodrugs of compounds of the invention are included within the scope of the invention. Examples of pro-drug functionality suitable for the compounds of the present invention aredescribed in Drugs of Today, Volume 19, Number 9, 1983, pp 499 – 538 and in Topics inChemistry, Chapter 31, pp 306 – 316 and in “Design of Prodrugs” by H. Bundgaard, Elsevier,1985, Chapter 1 (the disclosures in which documents are incorporated herein by reference). It will further be appreciated by those skilled in the art, that certain moieties, known to those skilled in the art as “pro-moieties”, for example as described by H. Bundgaard in “Design of Prodrugs” (the disclosure in which document is incorporated herein by reference) may be placed on appropriate functionalities when such functionalities are present within compounds of the invention. Also included within the scope of the compound and various salts of the invention are polymorphs thereof. Compounds of formula (I) may exist in a number of different geometric isomeric, and tautomeric forms and references to compounds of formula (I) include all such forms. For the avoidance of doubt, where a compound can exist in one of several geometric isomeric or CEN-C-P3772PCT tautomeric forms and only one is specifically described or shown, all others are nevertheless embraced by formula (I). The present invention includes all pharmaceutically acceptable isotopically-labelled compounds of the invention, i.e. compounds of formula (I), wherein one or more atoms are replaced by atoms having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes suitable for inclusion in the compounds of the invention compriseisotopes of hydrogen, such as 2H (D) and 3H (T), carbon, such as 11C, 13C and 14C, fluorine,such as18F, nitrogen, such as13N and15N, oxygen, such as15O,17O and18O. Certain isotopically-labelled compounds of formula (I), for example, those incorporating a radioactive isotope, are useful in drug and / or substrate tissue distribution studies. The compounds of formula (I) can also have valuable diagnostic properties in that they can be used for detecting or identifying the formation of a complex between a labelled compound and other molecules, peptides, proteins, enzymes or receptors. The detecting or identifying methods can use compounds that are labelled with labelling agents such as radioisotopes, enzymes, fluorescent substances, luminous substances (for example, luminol, luminol derivatives, luciferin, aequorin and luciferase), etc. The radioactive isotopes tritium, i.e.3H (T), and carbon-14, i.e.14C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection.Substitution with heavier isotopes such as deuterium, i.e.2H (D), may afford certaintherapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances.Substitution with positron emitting isotopes, such as 11C, 18F, 15O and 13N, can be useful inPositron Emission Topography (PET) studies for examining target occupancy.Isotopically-labelled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Preparations using appropriate isotopically-labelled reagents in place of the non-labelled reagent previously employed. Methods for the Preparation of Compounds of Formula (I) CEN-C-P3772PCT In this section, as in all other sections of this application unless the context indicates otherwise, references to formula (I) also include all other sub-groups and examples thereof as defined herein.The compounds pertaining to the invention described herein may be prepared in a stepwisesynthetic sequence as illustrated in the Schemes below. Compounds of formula (I) can be prepared in accordance with synthetic methods well known to the skilled person. For example, one skilled in the art will appreciate that the chemical steps and choice of protecting groups may be managed in any order to enable synthetic success. According to a further aspect of the invention there is provided a process for preparing a compound of formula (I) as hereinbefore defined which comprises: (a) reacting a compound of formula (II): wherein F, S2, Y2, R1, Y1and S1are as defined hereinbefore, with a compound of formula where Z1, Z3, Z4 and Z5 are as defined hereinbefore, Z2 is either as defined hereinbefore ormay represent an appropriately protected derivative of Z2, such as Z2 may contain aprotecting group, such as Boc, wherein Boc represents an amine protecting group which istertbutoxycarbonyl; followed by CEN-C-P3772PCT(b) deprotecting the Boc protecting groups of the product of step (a) to prepare acompound of formula (I). Step (a) typically comprises reacting the compound of formula (II) with the compound offormula (III) in the presence of triethylamine, DMF and HATU. Detailed experimental for step(a) is described in Example 1, Step 1 herein.Step (b) typically comprises TFA in DCM and water. Detailed experimental for step (b) isdescribed in Example 1, Step 2 herein.Compounds of formula (II) may be prepared according to the methods described inIntermediates 1 to 5 herein.Compounds of formula (III) may be prepared according to the methods described inIntermediates A to D herein.It will be appreciated that the intermediate compounds of formula (II) and (III) constitute novelcompounds and therefore form an additional aspect of the invention. Thus, according to a further aspect of the invention, there is provided a compound of formula (II) or a compound offormula (III) as defined herein. According to a further aspect of the invention, there is provideda compound of formula (II): wherein F, S2, Y2, R1, Y1and S1are as defined herein, or a compound of formula (III): CEN-C-P3772PCT wherein Z3, Z4 and Z5 are as defined herein, Z1 represents benzyl optionally substituted byone or more halogen or haloC1-6alkyl groups, and Z2is either as defined in claim 1 or may represent an appropriately protected derivative of Z2, such as Z2may contain a protecting group, such as Boc, wherein Boc represents an amine protecting group which is tertbutoxycarbonyl. Pharmaceutical CompositionsWhile it is possible for the compound of formula (I) to be administered alone, it is preferableto present it as a pharmaceutical composition (e.g. formulation).Thus, according to a further aspect, the invention provides a pharmaceutical composition,and methods of making a pharmaceutical composition comprising (e.g. admixing) at leastone compound of the invention where L represents a cationic anti-microbial peptide, together with one or more pharmaceutically acceptable excipients and optionally other therapeutic or prophylactic agents, as described herein. The pharmaceutically acceptable excipient(s) can be selected from, for example, carriers (e.g. a solid, liquid or semi-solid carrier), adjuvants, diluents, fillers or bulking agents, granulating agents, coating agents, release-controlling agents, binding agents, disintegrants, lubricating agents, preservatives, antioxidants, buffering agents, suspending agents, thickening agents, flavouring agents, sweeteners, taste masking agents, stabilisers or any other excipients conventionally used in pharmaceutical compositions. Examples of excipients for various types of pharmaceutical compositions are set out in more detail below. CEN-C-P3772PCT The term “pharmaceutically acceptable” as used herein pertains to compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject (e.g. human) without excessivetoxicity (i.e. generally recognised as safe (GRAS)), irritation, allergic response, or otherproblem or complication, commensurate with a reasonable benefit / risk ratio. Each carrier, excipient, etc. must also be “acceptable” in the sense of being compatible with the other ingredients of the formulation. Pharmaceutical compositions containing compounds of the invention can be formulated in accordance with known techniques, see for example, Remington’s Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, USA. The pharmaceutical compositions can be in any form suitable for parenteral, intranasal, intrabronchial, sublingual, ophthalmic, otic, rectal, intra-vaginal, or transdermal administration. Where the compositions are intended for parenteral administration, they can be formulated for intravenous, intramuscular, intraperitoneal, subcutaneous administration or for direct delivery into a target organ or tissue by injection, infusion or other means of delivery. The delivery can be by bolus injection, short term infusion or longer-term infusion and can be via passive delivery or through the utilisation of a suitable infusion pump or syringe driver. Pharmaceutical formulations adapted for parenteral administration include aqueous and non- aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats,co-solvents, surface active agents, organic solvent mixtures, cyclodextrin complexation agents,emulsifying agents (for forming and stabilizing emulsion formulations), liposome components for forming liposomes, gellable polymers for forming polymeric gels, lyophilisation protectants andcombinations of agents for, inter alia, stabilising the active ingredient in a soluble form andrendering the formulation isotonic with the blood of the intended recipient. Pharmaceutical formulations for parenteral administration may also take the form of aqueous and non- aqueous sterile suspensions which may include suspending agents and thickening agents (R. G. Strickly, Solubilizing Excipients in oral and injectable formulations, Pharmaceutical Research, Vol 21(2) 2004, p 201-230). The formulations may be presented in unit-dose or multi-dose containers, for example sealed ampoules, vials and prefilled syringes, and may be stored in a freeze-dried (lyophilised) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. CEN-C-P3772PCT The pharmaceutical formulation can be prepared by lyophilising a compound of the invention. Lyophilisation refers to the procedure of freeze-drying a composition. Freeze- drying and lyophilisation are therefore used herein as synonyms. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules and tablets. Pharmaceutical compositions of the present invention for parenteral injection can also comprise pharmaceutically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as sunflower oil, safflower oil, corn oil or olive oil), and injectable organic esters such as ethyloleate. Proper fluidity can be maintained, for example, by the use of thickening or coatingmaterials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. The compositions of the present invention may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various anti-bacterial and antifungal agents, for example, paraben, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include agents to adjust tonicity such as sugars, sodium chloride, and the like. Prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents which delay absorption such as aluminium monostearate and gelatin. In one preferred embodiment of the invention, the pharmaceutical composition is in a form suitable for i.v. administration, for example by injection or infusion. For intravenous or subcutaneous administration, the solution can be dosed as is, or can be injected into an infusion bag (containing a pharmaceutically acceptable excipient, such as 0.9% saline or 5% dextrose), before administration. CEN-C-P3772PCT In another preferred embodiment, the pharmaceutical composition is in a form suitable for subcutaneous (s.c.) administration. The compound of the invention may be formulated with a carrier and administered in the form of nanoparticles, the increased surface area of the nanoparticles assisting their absorption. In addition, nanoparticles offer the possibility of direct penetration into the cell. Nanoparticle drug delivery systems are described in “Nanoparticle Technology for Drug Delivery”, edited by Ram B Gupta and Uday B. Kompella, Informa Healthcare, ISBN 9781574448573, published 13thMarch 2006. Nanoparticles for drug delivery are also described in J. Control. Release, 2003, 91 (1-2), 167-172, and in Sinha et al.,Mol. CancerTher. August 1, (2006) 5, 1909.The pharmaceutical compositions typically comprise from approximately 1% (w / w) to approximately 95% (w / w) active ingredient and from 99% (w / w) to 5% (w / w) of a pharmaceutically acceptable excipient or combination of excipients. Preferably, the compositions comprise from approximately 20% (w / w) to approximately 90%(w / w) active ingredient and from 80% (w / w) to 10% of a pharmaceutically acceptable excipient or combination of excipients. The pharmaceutical compositions comprise from approximately1% to approximately 95%, preferably from approximately 20% to approximately 90%, activeingredient. Pharmaceutical compositions according to the invention may be, for example, in unit dose form, such as in the form of ampoules, vials, suppositories, pre-filled syringes, dragées, tablets or capsules. The pharmaceutically acceptable excipient(s) can be selected according to the desiredphysical form of the formulation and can, for example, be selected from diluents (e.g. soliddiluents such as fillers or bulking agents; and liquid diluents such as solvents and co- solvents), disintegrants, buffering agents, lubricants, flow aids, release controlling (e.g. release retarding or delaying polymers or waxes) agents, binders, granulating agents, pigments, plasticizers, antioxidants, preservatives, flavouring agents, taste masking agents, tonicity adjusting agents and coating agents. The skilled person will have the expertise to select the appropriate amounts of ingredientsfor use in the formulations. For example, tablets and capsules typically contain 0-20%disintegrants, 0-5% lubricants, 0-5% flow aids and / or 0-99% (w / w) fillers / or bulking agents (depending on drug dose). They may also contain 0-10% (w / w) polymer binders, 0-5% (w / w) antioxidants, 0-5% (w / w) pigments. Slow release tablets would in addition contain 0-99% (w / w) release-controlling (e.g. delaying) polymers (depending on dose). The film coats of the CEN-C-P3772PCT tablet or capsule typically contain 0-10% (w / w) polymers, 0-3% (w / w) pigments, and / or 0-2% (w / w) plasticizers. Parenteral or subcutaneous formulations typically contain 0-20% (w / w) buffers, 0-50% (w / w) co-solvents, and / or 0-99% (w / w) Water for Injection (WFI) (depending on dose and if freeze dried). Formulations for intramuscular depots may also contain 0-99% (w / w) oils. The compounds of the invention can also be formulated as solid dispersions. Solid dispersions are homogeneous extremely fine disperse phases of two or more solids. Solid solutions (molecularly disperse systems), one type of solid dispersion, are well known for use in pharmaceutical technology (see (Chiou and Riegelman, J. Pharm. Sci., 60, 1281- 1300 (1971)) and are useful in increasing dissolution rates and increasing the bioavailability of poorly water-soluble drugs. The pharmaceutical formulations may be presented to a patient in “patient packs” containing an entire course of treatment in a single package, usually a blister pack. Patient packs have an advantage over traditional prescriptions, where a pharmacist divides a patient’s supply ofa pharmaceutical from a bulk supply, in that the patient always has access to the packageinsert contained in the patient pack, normally missing in patient prescriptions. The inclusion of a package insert has been shown to improve patient compliance with the physician’sinstructions. One example of a patient pack includes a prefilled syringe. Such pre-filledsyringes already contain the drug substance. The front-end portion of a pre-filled syringe to which a needle is to be attached is sealed with a nozzle cap. Prior to injection, the nozzle cap is removed from the front-end portion and a needle is attached thereto. A gasket is then slid by pushing a plunger rod toward the front-end portion so that the drug is expelled. Compositions for nasal delivery include ointments, creams, sprays, patches, gels, liquid drops and inserts (for example intraocular inserts). Such compositions can be formulated in accordance with known methods. Examples of formulations for rectal or intra-vaginal administration include pessaries and suppositories which may be, for example, formed from a shaped moldable or waxy material containing the active compound. Solutions of the active compound may also be used for rectal administration. Compositions for administration by inhalation may take the form of inhalable powder compositions or liquid or powder sprays, and can be administrated in standard form using CEN-C-P3772PCT powder inhaler devices or aerosol dispensing devices. Such devices are well known. For administration by inhalation, the powdered formulations typically comprise the active compound together with an inert solid powdered diluent such as lactose. The compound of the invention will generally be presented in unit dosage form and, as such, will typically contain sufficient compound to provide a desired level of biological activity. For example, a formulation may contain from 1 nanogram to 2 grams of active ingredient, e.g. from 1 nanogram to 2 milligrams of active ingredient. Within these ranges, particular sub- ranges of compound are 0.1 milligrams to 2 grams of active ingredient (more usually from 10 milligrams to 1 gram, e.g.50 milligrams to 500 milligrams), or 1 microgram to 20 milligrams (for example 1 microgram to 10 milligrams, e.g.0.1 milligrams to 2 milligrams of active ingredient). The active compound will be administered to a patient in need thereof (for example a human or animal patient) in an amount sufficient to achieve the desired therapeutic effect. Therapeutic Uses According to a further aspect of the invention, there is provided a compound of formula (I) as defined herein for use in therapy. According to a further aspect of the invention, there is provided a compound of formula (I) as defined herein for use in the treatment of a disease or disorder mediated and / or caused by an infective agent. According to a further aspect of the invention, there is provided the use of a compound of formula (I) as defined herein in the manufacture of a medicament for use in the treatment of a disease or disorder mediated and / or caused by an infective agent.According to a further aspect of the invention, there is provided a method of treating adisease or disorder mediated and / or caused by an infective agent which comprisesadministering to an individual in need thereof a compound of formula (I) as defined herein. Examples of infective agents include any pathogen such as a bacteria, fungus, parasite or virus. Thus, in one embodiment, the disease or disorder mediated by and / or caused by an infective agent is bacterial infection. CEN-C-P3772PCT Examples of such as bacterial infection include infection by the following bacteria:Staphylococcus sp. such as Staphylococcus aureus (including methicillin resistantStaphylococcus aureus (MRSA)), Clostridia sp (e.g. Clostridium difficile, Clostridium tetaniand Clostridium botulinum), Enterobacter species, Mycobacterium tuberculosis, Shigella sp.such as Shigelladysenteriae, Campylobacter sp. such as Campylobacter jejuni, Enterococcus sp. such as Enterococcus faecalis, Bacillus anthracis, Yersinia pestis, Bordetella pertussis, Streptococcal species, Salmonella thyphimurim, Salmonella enterica, Chlamydia species, Treponemapallidum, Neisseria gonorrhoeae, Borreliaburgdorferi, Vibriocholerae, Corynebacterium diphtheriae, Helicobacter pylori, Gram-negative pathogens, suchas Acinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella pneumoniae, andEscherichia coli (and including strains that are resistant to one or more classes of anti-biotics, especially multi-drug resistant (MDR) strains). In view of the fact that Polymyxinderivatives (such as those present within the compounds of the invention) are typically selective for Gram-negative bacteria, it will be appreciated that the present invention finds particular utility in treating bacterial infection by Gram-negative pathogens, such asAcinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella pneumoniae, andEscherichia coli (and including strains that are resistant to one or more classes of anti-biotics, especially multi-drug resistant (MDR) strains).The compound of the invention is generally administered to a subject in need of suchadministration, for example a human or animal patient, preferably a human.The compound of the invention will typically be administered in amounts that aretherapeutically or prophylactically useful and which generally are non-toxic. However, in certain situations (for example in the case of life threatening diseases), the benefits ofadministering a compound of the invention may outweigh the disadvantages of any toxiceffects or side effects, in which case it may be considered desirable to administer acompound of the invention in amounts that are associated with a degree of toxicity.The compound of the invention may be administered over a prolonged term (i.e. chronicadministration) to maintain beneficial therapeutic effects or may be administered for a shortperiod only (i.e. acute administration). Alternatively, they may be administered in acontinuous manner or in a manner that provides intermittent dosing (e.g. a pulsatile manner).A typical daily dose of the compound of the invention can be in the range from 100picograms to 100 milligrams per kilogram of body weight, more typically 5 nanograms to 25 milligrams per kilogram of bodyweight, and more usually 10 nanograms to 15 milligrams per CEN-C-P3772PCT kilogram (e.g.10 nanograms to 10 milligrams, and more typically 1 microgram per kilogram to 20 milligrams per kilogram, for example 1 microgram to 10 milligrams per kilogram) per kilogram of bodyweight although higher or lower doses may be administered where required.The compound of the invention can either be administered on a daily basis or on a repeatbasis every 2, or 3, or 4, or 5, or 6, or 7, or 10 or 14, or 21, or 28 days for example.Alternatively, the compound of the invention can be administered by infusion, multiple timesper day.The compound of the invention may be administered in a range of doses, for example 1 to1500 mg, 2 to 800 mg, or 5 to 500 mg, e.g.2 to 200 mg or 10 to 1000 mg, particularexamples of doses including 10, 20, 50 and 80 mg. The compound of the invention may beadministered once or more than once each day. The compound of the invention can beadministered continuously (i.e. taken every day without a break for the duration of thetreatment regimen). Alternatively, the compound of the invention can be administeredintermittently (i.e. taken continuously for a given period such as a week, then discontinued for a period such as a week and then taken continuously for another period such as a week and so on throughout the duration of the treatment regimen). Examples of treatment regimens involving intermittent administration include regimens wherein administration is in cycles of one week on, one week off; or two weeks on, one week off; or three weeks on, one week off; or two weeks on, two weeks off; or four weeks on two weeks off; or one week onthree weeks off - for one or more cycles, e.g.2, 3, 4, 5, 6, 7, 8, 9 or 10 or more cycles.In one particular dosing schedule, a patient will be given an infusion of a compound of theinvention for periods of one hour daily for up to ten days in particular up to five days for oneweek, and the treatment repeated at a desired interval such as two to four weeks, in particular every three weeks.More particularly, a patient may be given an infusion of a compound of the invention forperiods of one hour daily for 5 days and the treatment repeated every three weeks. In another particular dosing schedule, a patient is given an infusion over 30 minutes to 1 hour followed by maintenance infusions of variable duration, for example 1 to 5 hours, e.g.3 hours. In a further particular dosing schedule, a patient is given a continuous infusion for a period of12 hours to 5 days, and in particular a continuous infusion of 24 hours to 72 hours. CEN-C-P3772PCTUltimately, however, the quantity of compound of the invention administered and the type ofcomposition used will be commensurate with the nature of the disease or physiological condition being treated and will be at the discretion of the physician.It will be appreciated that the compound of the invention can be used as a single agent or incombination with other therapeutic agents. Combination experiments can be performed, forexample, as described in Chou TC, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regulat 1984;22: 27–55.Where the compound of the invention is administered in combination therapy with one, two,three, four or more other therapeutic agents (preferably one or two, more preferably one),the agents can be administered simultaneously or sequentially. In the latter case, the two ormore agents will be administered within a period and in an amount and manner that issufficient to ensure that an advantageous or synergistic effect is achieved. When administered sequentially, they can be administered at closely spaced intervals (for example over a period of 5-10 minutes) or at longer intervals (for example 1, 2, 3, 4 or more hoursapart, or even longer periods apart where required), the precise dosage regimen beingcommensurate with the properties of the therapeutic agent(s). These dosages may be administered for example once, twice or more per course of treatment, which may be repeated for example every 7, 14, 21 or 28 days. It will be appreciated that the preferred method and order of administration and the respective dosage amounts and regimes for each component of the combination will dependon the particular other medicinal agent and compound of the invention being administered,their route of administration, the particular tumour being treated and the particular host being treated. The optimum method and order of administration and the dosage amounts and regime can be readily determined by those skilled in the art using conventional methods and in view of the information set out herein.The weight ratio of the compound of the invention and the one or more other therapeuticagent(s) when given as a combination may be determined by the person skilled in the art. Said ratio and the exact dosage and frequency of administration depends on the particularcompound of the invention and the other therapeutic agent(s) used, the particular conditionbeing treated, the severity of the condition being treated, the age, weight, gender, diet, time of administration and general physical condition of the particular patient, the mode of administration as well as other medication the individual may be taking, as is well known to CEN-C-P3772PCT those skilled in the art. Furthermore, it is evident that the effective daily amount may be lowered or increased depending on the response of the treated subject and / or depending onthe evaluation of the physician prescribing the compound of present invention. A particularweight ratio for the compound of the invention and another therapeutic agent may rangefrom 1 / 10 to 10 / 1, more in particular from 1 / 5 to 5 / 1, even more in particular from 1 / 3 to 3 / 1. EXAMPLES The invention will now be illustrated, but not limited, by reference to the specific embodiments described in the following examples. Compounds are named using an automated naming package (ChemDraw) or are as named by the chemical supplier. The following synthetic procedures are provided for illustration of the methods used; for a given preparation or step the precursor used may not necessarily derive from the individual batch synthesised according to the step in the description given. Analytical Methods Wherein examples and preparations cite analytical data, the following analytical methods were used unless otherwise specified: NMR1H NMR spectra were recorded at room temperature on a JEOL ECZ400S / L1 or JEOL 400YH spectrometer (400 MHz). Data are presented as follows: chemical shift in ppm, integration, multiplicity (br = broad, s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, dd = doublet of doublet) and coupling constant in Hz. Solvents used for samples are specified in the specific experimental procedures for each compound.UPLC-MS (Method A)Instrument: Waters Acquity UPLC H-Class system; Column: Acquity CSH C181.7 μm 2.1 x50 mm (cat.186005296 from Waters). Injection volume: 0.5 µL. Eluent A: 5% MeCN in water (volume), eluent B: 5% water in MeCN (volume), both eluents contained 0.1% (volume) formic acid. Gradient: 0-0.2 min 0% B, 0.2-3.5 min gradient 0% to 100% B, 3.5 to 4.5 min 100%. Flow 0.4 mL.min-1; temperature: 40 °C; photodiode array: 215-350 nm. UPLC-MS (Method B) UPLC-MS analysis was carried out on a Waters Acquity UPLC system consisting of an Acquity I-Class Sample Manager-FL, Acquity I-Class Binary Solvent Manager and an Acquity UPLC Column Manager. UV detection was afforded using an Acquity UPLC CEN-C-P3772PCT PDA detector (scanning from 210 to 400 nm), whilst mass detection was achievedusing an Acquity QDa detector (mass scanning from 100–1250 Da; positive andnegative modes simultaneously), and ELS detection was achieved using an Acquity UPLC ELS Detector.Acidic 2min: 0.1% v / v formic acid in 10mM ammonium formate [Eluent A]; 0.1% v / v formicacid in MeCN [Eluent B]; flow rate 0.8mL / min; column oven 50˚C; sample manager 20˚C; injection volume 2µL and 1.5 minutes equilibration time between samples on a Waters Acquity UPLC BEH C18 column (2.1 × 50 mm, 1.7 µm). Time (min) Eluent A (%) Eluent B (%)0.00 95 50.25 95 51.25 5 951.55 5 951.65 95 52.00 95 5Basic 2 min: As UPLC method B using 0.1% ammonia in 10 mM ammonium bicarbonate[Eluent A]; 0.1% ammonia in MeCN [Eluent B]; flow rate 0.8mL / min; column oven 50˚C; sample manager 20˚C; injection volume 2μL and 1.5 minutes equilibration time between samples on a Waters Acquity UPLC BEH C18 column (2.1 × 50 mm, 1.7 μm). UPLC Method C UPLC-MS analysis was carried out on a Waters Acquity UPLC system consisting of an Acquity I-Class Sample Manager-FL, Acquity I-Class Binary Solvent Manager and an Acquity UPLC Column Manager. UV detection was afforded using an Acquity UPLC PDA detector (scanning from 210 to 400 nm), whilst mass detection was achieved using an Acquity QDa detector (mass scanning from 100–1250 Da; positive and negative modes simultaneously), and ELS detection was achieved using an Acquity UPLC ELS Detector. UPLC conditions: 0.1% ammonia in 10 mM ammonium bicarbonate [Eluent A]; 0.1% ammoniain MeCN [Eluent B]; flow rate 0.8mL / min; column oven 50˚C; sample manager 20˚C; injection CEN-C-P3772PCT volume 2μL and 1.5 minutes equilibration time between samples on a Waters Acquity UPLC BEH C18 column (2.1 × 50 mm, 1.7 μm). Time (min) Eluent A (%) Eluent B (%) 0.00 95 5 0.25 95 5 2.75 5 95 3.25 5 95 3.35 95 5 4.00 95 5UPLC Method D (TFA 4 min)UPLC-MS analysis was carried out on a Waters Acquity UPLC system consisting ofan Acquity I-Class Sample Manager-FL, Acquity I-Class Binary Solvent Manager andan Acquity UPLC Column Manager. UV detection was afforded using an Acquity UPLC PDA detector (scanning from 210 to 400 nm), whilst mass detection was achieved using an Acquity QDa detector (mass scanning from 100–1250 Da; positive and negative modes simultaneously), and ELS detection was achieved using an Acquity UPLC ELS Detector. UPLC conditions: 0.05% v / v trifluoroacetic acid in water [Eluent A]; 0.05% v / v trifluoroacetic acid in MeCN [Eluent B]; flow rate 0.8mL / min; column oven 50˚C; sample manager 20˚C; injection volume 2μL and 1.5 minutes equilibration time between samples, on a Waters Acquity UPLC CSH C18 column (2.1 × 50 mm, 1.7 μm). Time (min) Eluent A (%) Eluent B (%)0.00 95 50.25 95 52.75 5 953.25 5 953.35 95 54.00 95 5Abbreviations Wherein the following abbreviations have been used, the following meanings apply: CEN-C-P3772PCT Boc is tert-butyloxycarbonyl; Cbz is benzyloxycarbonyl; CV is column volume; d is doublet; Dab is 2,4-diaminobutyric acid; Dap is 2,3-diaminopropionic acid; DCM is dichloromethane; DMF is dimethylformamide; DMSO is dimethylsulfoxide; DMSO-D6 is deuterated DMSO; ESI is electrospray ionisation technique; eq. is equivalent; EtOH is ethanol; EtOAc is ethyl acetate; FITC is Fluorescein Isothiocyanate; FMOC is 9-Fluorenylmethyloxycarbonyl; g is gram; Gly is glycine; HATU is O-(7-azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate; L is litre; Leu is leucine; m is multiplet; mg is milligram; M is molar; Me is methyl; MeCN is acetonitrile; MeOH is methanol; MeOD is deuteriomethanol; MHz is megaHertz; mL is millilitre; mmol is millimole; MS is mass spectrometry; MTBE is methyl tert butyl ether; NaHCO3 is sodium hydrogen carbonate; NH3 is ammonia; NMR is nuclear magnetic resonance; Phe is phenylalanine; CEN-C-P3772PCT Ppm is parts per million;RT is retention time;Ruphos Pd G3 is (2-Dicyclohexylphosphino-2′,6′-diisopropoxy-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate; s is singlet; t is triplet; Thr is threonine; TFA is trifluoroacetic acid; µL is microlitre; UPLC is ultra performance liquid chromatography; and v is volume. Preparation of Intermediates Intermediate A H-Phe(4-F)-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)- Dab(Boc)-Thr](Step 1) Cbz-Dab(Boc)-Thr-CO2Me (2S)-2-(Benzyloxycarbonylamino)-4-(tert-butoxycarbonylamino)butanoic acid (44.9 g, 128 mmol, 1 eq.) was dissolved in N,N-dimethylformamide (100 mL) and dichloromethane (50 mL). Methyl (2S,3R)-2-amino-3-hydroxy-butanoate hydrochloride (26.3 g, 147 mmol, 1.15 eq.) was dissolved in N,N-dimethylformamide (50 mL) and dichloromethane (70 mL).Triethylamine (39.9 g, 395 mmol, 55.0 mL, 3.1 eq.) was then added slowly, forming a thick white suspension, which was then diluted with dichloromethane (180 mL).1- [Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (97.8 g, 257 mmol, 2 eq.) was added in portions over 20 minutes while cooling the reaction in an ice bath. Following addition, the reaction mixture was allowed to warm to room temperature and stirred for 3 hours. The reaction mixture was partitioned between ethyl acetate (1 L) and aqueous hydrochloric acid (1M, 500 mL). The layers were separated, and the aqueous layer extracted with ethyl acetate (2 x 1 L) and the combined CEN-C-P3772PCT organic layers were washed with saturated aqueous sodium bicarbonate (4 x 900 mL) and brine (3 x 1.2 L). The solution was dried over magnesium sulfate, filtered and the solvent removed under vacuum. The resultant yellow gum was left overnight in a vacuum oven at80°C to yield the title product as a yellow gum (89.3 g).1H NMR (396 MHz, DMSO-d6): δ [ppm] = 7.97 (d, J = 8.5 Hz, 1H), 7.49 (d, J = 7.9 Hz, 1H), 7.37-7.27 (m, 5H), 6.82 (t, J = 5.1 Hz, 1H), 5.07 (d, J = 4.8 Hz, 1H), 5.00 (s, 2H), 4.29 (dd, J = 8.5, 3.0 Hz, 1H), 4.19-4.11 (m, 2H), 3.61 (s, 3H), 3.12-2.93 (m, 2H), 1.79 (td, J = 13.3, 6.9 Hz, 1H), 1.59 (td, J = 14.2, 6.7 Hz, 1H), 1.37 (s, 9H), 1.05 (d, J = 6.7 Hz, 3H).(Step 2) Cbz-Dab(Boc)-Thr-CO2H Cbz-Dab(Boc)-Thr-CO2Me (Step 1) (89.3 g, 114 mmol, 1 eq.) was dissolved intetrahydrofuran (100 mL) and water (100 mL). Lithium hydroxide, monohydrate (6.30 g, 150 mmol, 1.3 eq.) was dissolved in water (150 mL) and added slowly to the stirring solution. The reaction was left to stir for 80 mins. The reaction mixture was concentrated under vacuum and tert-butyl methyl ether (500 mL) was added. The mixture was acidified to pH ~6 withaqueous hydrochloric acid (4 M). The phases were separated, and to the aqueous phasetert-butyl methyl ether (500 mL) was added. The aqueous layer was acidified to pH ~3 with aqueous hydrochloric acid (4 M). The phases were again separated and the aqueous was extracted with tert-butyl methyl ether (500 mL). The combined organic layers were washed with brine (300 mL). The solvent was removed under vacuum, to furnish the title compound as an amorphouswhite solid (59.8 g).UPLC-MS (Method A): (ESIneg): m / z = [M - H]- 452.2.1H NMR (396 MHz, DMSO-d6): δ [ppm] = 12.52 (s, 1H), 7.80 (d, J = 8.5 Hz, 1H), 7.49 (d, J = 7.9 Hz, 1H), 7.37-7.27 (m, 5H), 6.82 (t, J = 5.1 Hz, 1H), 5.01 (s, 2H), 4.21-4.11 (m, 3H), 3.05- 2.94 (m, 2H), 1.80 (td, J = 13.2, 7.3 Hz, 1H), 1.59 (td, J = 14.4, 6.1 Hz, 1H), 1.45-1.26 (s, 9H), 1.04 (d, J = 6.1 Hz, 3H).(Step 3) Cbz-Dab(Boc)-Thr-Dap(Boc)-OMe CEN-C-P3772PCT CBz-Dab(Boc)-Thr-CO2H (Step 2) (51.8 g, 114 mmol, 1 eq.) was dissolved in N,N- dimethylformamide (75 mL) and dichloromethane (200 mL). Methyl (2S)-2-amino-3-(tert- butoxycarbonylamino)propanoate (30.1 g, 140 mmol, 1.2 eq.) was dissolvedin dichloromethane (250 mL) and added to the reaction mixture. Triethylamine (34.9 g, 344mmol, 48.0 mL, 3 eq.) and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5- b]pyridinium 3-oxid hexafluorophosphate (76.2 g, 200 mmol, 1.8 eq.) were then added to the reaction mixture. The solution was stirred at room temperature for 2 hr. Additional N,N- dimethylformamide (50 mL) was added and the solution left to stir overnight. The solution was concentrated under vacuum and partitioned between tert-butyl methyl ether (1 L) and aqueous hydrochloric acid (1N, 500 mL). The layers were separated and the aqueous extracted with tert-butyl methyl ether (1 L, 2 x 750 mL). The combined organic layers were washed with saturated aqueous sodium bicarbonate (3 x 1.2L), brine (3 x 900 mL), and dried over magnesium sulfate. The solids were filtered and washed with ethyl acetate and tert- butyl methyl ether. The solution was concentrated under vacuum to yield the title product asa pale orange-pink solid (93.3 g).UPLC-MS (Method A): MS (ESIPos): m / z = [M - Boc + H]+ 554.3.1H NMR (396 MHz, DMSO-d6): δ [ppm] = 8.01 (d, J = 7.5 Hz, 1H), 7.73 (d, J = 8.1 Hz, 1H), 7.52 (d, J = 7.9 Hz, 1H), 7.35-7.29 (m, 5H), 6.87 (t, J = 5.7 Hz, 1H), 6.77 (t, J = 4.7 Hz, 1H), 5.01 (s, 2H), 4.34-4.29 (m, 1H), 4.22-4.19 (m, 1H), 4.11-4.06 (m, 1H), 3.95-3.92 (m, 1H), 3.58(s, 3H), 3.29-3.23 (m, 2H), 3.01-2.94 (m, 3H), 1.80 (dd, J = 13.5, 6.4 Hz, 1H), 1.60 (q, J = 7.3Hz, 1H), 1.36 (d, J = 3.2 Hz, 18H), 1.04 (d, J = 6.2 Hz, 3H).(Step 4) Cbz-Dab(Boc)-Thr-Dap(Boc)-OH Cbz-Dab(Boc)-Thr-Dap(Boc)-OMe (Step 3) (28.0 g, 42.8 mmol, 1 eq.) was dissolved intetrahydrofuran (300 mL), and a solution of lithium hydroxide monohydrate (1.89 g, 45.0mmol, 1.05 eq.) in water (100 mL) was added. This was stirred for 30 min. The CEN-C-P3772PCT tetrahydrofuran was removed under reduced pressure and the remaining aqueous fraction diluted with water (50 mL) and ethyl acetate (200 mL). The stirring solution was acidified to pH 3 with aqueous hydrochloric acid (2 M). The organic phase separated, and the aqueous phase extracted with ethyl acetate (100 mL). The combined organic layers were dried over magnesium sulfate, filtered, and evaporated under reduced pressure to give the title product as an off white solid (30.0 g).UPLC-MS (Method A): MS (ESIneg): m / z = [M - H]- 638.3.1H NMR (396 MHz, MeOD-d4): δ [ppm] = 7.35-7.25 (m, 5H), 5.08 (s, 2H), 4.48-4.45 (m, 1H), 4.31 (d, J = 3.6 Hz, 1H), 4.21-4.17 (m, 2H), 3.54 (dd, J = 14.2, 3.9 Hz, 1H), 3.42-3.36 (m, 1H), 3.19-3.08 (m, 2H), 1.99-1.95 (m, 1H), 1.78-1.73 (m, 1H), 1.41-1.37 (m, 18H), 1.19-1.16 (m, 3H).(Step 5) Cbz-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] CBz-Dab(Boc)-Thr-Dap(Boc)-OH (Step 4) (1.10 eq, 12.72 g, 19.9 mmol) and (BOC)3Polymyxin B heptapeptide (prepared according to Li et al, Synthesis 2015, 47, 2088–2092)(1.10 eq, 12.72 g, 19.9 mmol) (1.00 eq, 19.20 g, 18.1 mmol) were dissolved in DCM(1.28L) and DMF (320mL), N,N-Diisopropylethylamine (4.00 eq, 13 mL, 72.3 mmol) wasthen added and the solution allowed to stir for 10 minutes before adding O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (1.20 eq, 8.25 g, 21.7 mmol) . The reaction was allowed to stir overnight. The reaction was concentrated to a thick oil, water (500 mL) was added, and the resulting mixture allowed tostir for 2 hours, water (500 mL) was added and the solid allowed to digest overnight. Themixture was filtered and the white solid dried to constant mass product 1 (27.50 g,16.3mmol, 90 % yield).UPLC-MS (Method B): m / z 843.0 [M+2H]2+ CEN-C-P3772PCT1HNMR (400 MHz, DMSO-D6) δ 8.68 – 8.56 (m, 2H), 8.57 – 8.33 (m, 2H), 8.17 (d, J = 8.7Hz, 2H), 8.04 (s, 5H), 7.99 – 7.79 (m, 4H), 7.76 (d, J = 8.5 Hz, 1H), 7.62 – 7.37 (m, 3H), 7.37– 7.23 (m, 11H), 7.21 (d, J = 4.4 Hz, 7H), 7.14 (dt, J = 8.6, 4.1 Hz, 1H), 7.08 (s, 1H), 6.94 (s,1H), 6.88 – 6.70 (m, 1H), 6.70 – 6.51 (m, 2H), 5.13 – 4.75 (m, 7H), 4.48 – 4.00 (m, 4H), 4.00– 3.74 (m, 4H), 2.85 (s, 38H), 2.67 (d, J = 17.0 Hz, 4H), 1.73 (d, J = 15.4 Hz, 15H), 1.53 (dd,J = 34.0, 7.2 Hz, 6H), 1.43 – 1.28 (m, 114H), 1.28 – 1.11 (m, 5H), 0.99 (td, J = 6.9, 3.8 Hz,17H), 0.87 – 0.75 (m, 3H), 0.64 (dd, J = 16.1, 6.5 Hz, 12H).(Step 6) H-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] Palladium on activated carbon (10%) (1.98 eq, 2.30 g, 21.6 mmol) was slurried with water (50mL) into a solution of Cbz-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] (Step 5) (1.00 eq, 23.00 g, 10.9 mmol) in Ethanol (875mL), and theresulting mixture was stirred vigorously. The atmosphere was exchanged for hydrogen gasby backfilling from a balloon.After 23 h, the reaction mixture was filtered through celite eluting with ethanol (ca. 3 L). Thefiltrate was concentrated under reduced pressure to afford an off-white foam (ca.23 g). Thefoam was dissolved in DMF (70 mL) and precipitated with water (250 mL), stirring for 4 h. The water was filtered and any collected solid returned to the source flask using MeOH. The solution was concentrated to a brownish glassy solid (20 g). The solid was dissolved in chloroform (250 mL) and allowed to stir for 30 min. Heptane (500 mL) was added slowly over15 min, stirring for ca.20 min and allowed to settle for 1 h before decanting the liquid awayto afford the title compound, (18.50 g) as a white solid.UPLC-MS (Method B) m / z 773.9 [M-2H]2-, 775.8 [M+2H]2+. CEN-C-P3772PCT(Step 7) Cbz-Phe(4-F)-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab H-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] (Step 6) (1.00 eq, 18.70 g, 11.8 mmol) and (2S)-2-(benzyloxycarbonylamino)-3-(4-fluorophenyl)propanoic acid (1.20 eq, 4.50 g, 14.2 mmol) were dissolved in DMF(650mL), N,N-Diisopropylethylamine (6.00 eq, 12 mL, 70.9 mmol) was added followedby HATU (1.20 eq, 5.40 g, 14.2 mmol) and the yellow solution stirred overnight at roomtemperature. The reaction mixture was concentrated to 150 mL then water (200 mL) wasadded dropwise while stirring. The formed orange solid was then collected via suctionfiltration. This solid was then triturated with DCM (100 mL), then the isolated solid was triturated with a mixture of MeOH / MeCN 1:1 (200 mL) and collected by suction filtration anddried to give the title product (18.10 g) as a creamy solid.UPLC-MS (Method B, basic) m / z 923.5 [M-2H]2-1H NMR (400 MHz, DMSO-D6) δ 8.70 – 8.58 (m, 1H), 8.54 – 8.42 (m, 1H), 8.29 – 8.15 (m,2H), 8.11 – 8.02 (m, 1H), 8.02 – 7.96 (m, 1H), 7.96 – 7.85 (m, 2H), 7.48 (d, J = 8.6 Hz, 2H),7.37 – 7.14 (m, 15H), 7.07 (t, J = 8.8 Hz, 3H), 7.02 – 6.91 (m, 1H), 6.89 – 6.74 (m, 2H), 6.71– 6.52 (m, 3H), 5.08 (s, 1H), 4.98 – 4.86 (m, 3H), 4.50 – 4.13 (m, 10H), 4.03 – 3.93 (m, 1H),3.90 (d, J = 8.2 Hz, 1H), 3.88 – 3.80 (m, 1H), 3.23 – 3.18 (m, 1H), 3.14 – 2.93 (m, 11H), 2.84– 2.74 (m, 2H), 2.71 – 2.62 (m, 1H), 1.99 – 1.69 (m, 8H), 1.69 – 1.58 (m, 3H), 1.58 – 1.46(m, 3H), 1.44 – 1.31 (m, 58H), 1.30 – 1.15 (m, 2H), 1.03 (dd, J = 6.4, 2.5 Hz, 7H), 0.96 –0.80 (m, 1H), 0.70 (d, J = 6.6 Hz, 3H), 0.66 (d, J = 6.3 Hz, 3H).(Step 8) H-Phe(4-F)-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] (Intermediate A) CEN-C-P3772PCT To a solution of Cbz-Phe(4-F)-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu- Dab(Boc)-Dab(Boc)-Thr] (Step 7) (1.00 eq, 44.70 g, 23.4 mmol) in ethanol (1000mL) was added a suspension of palladium on activated carbon (10%) (2.01 eq, 5.00 g, 47.0 mmol) in water (100mL). The atmosphere was exchanged for nitrogen gas then from nitrogen to hydrogen by backfilling from a balloon and the reaction mixture was allowed to stir under an atmosphere of hydrogen at room temperature overnight. The reaction mixture was filtered through celite eluting with: ethanol (ca.1000 mL) The resulting solution was concentrated under reduced pressure to afford 36.7g of crude as a white solid. The obtained crude was split in two and purified by column chromatography on silica gel (2 x 220 g cartridge, dry loading on silica, using 1g of silica / 1g of material) using a gradient of MeOH (2 to 20%, v / v) in DCM. The fractions were monitored by TLC (10% MeOH in DCM) Rf = 0.45, using ninhydride as a stain and then combined. Solvent was evaporated under reduced pressure to provide the desired product 1 (18.70 g,11.8 mmol, 50% yield) as a white solid.UPLC-MS (Method C) m / z 776.0 [M+2H]2+1H NMR (400 MHz, DMSO-D6) δ 8.67 – 8.39 (m, 1H), 8.27 – 7.84 (m, 4H), 7.29 – 7.21 (m,5H), 7.21 – 7.15 (m, 1H), 7.15 – 6.90 (m, 1H), 6.90 – 6.75 (m, 2H), 6.72 – 6.54 (m, 3H), 5.07(d, J = 4.7 Hz, 1H), 4.90 (d, J = 5.5 Hz, 1H), 4.49 – 4.05 (m, 8H), 4.04 – 3.95 (m, 1H), 3.95 –3.81 (m, 2H), 3.17 – 2.94 (m, 10H), 2.94 – 2.86 (m, 2H), 2.85 – 2.71 (m, 2H), 1.97 – 1.60 (m,10H), 1.59 – 1.47 (m, 3H), 1.46 – 1.33 (m, 60H), 1.32 – 1.20 (m, 1H), 1.07 – 1.00 (m, 8H),0.98 – 0.79 (m, 1H), 0.71 (d, J = 6.7 Hz, 4H), 0.67 (d, J = 6.3 Hz, 3H).Intermediate B H-Phe(4-Cl)-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)- Dab(Boc)-Thr] CEN-C-P3772PCT H-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] (Intermediate 1 , step 6) (1.10 g, 0.710 mmol), Fmoc-4-chloro-phenylalanine (1.20 eq, 359 mg, 0.852 mmol) and N,N-Diisopropylethylamine (4.00 eq, 0.49 mL, 2.84 mmol) weredissolved in DMF (30mL) and the resulting mixture stirred for 5 min. The solution was clearat this time. O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate(HATU) (1.50 eq, 405 mg, 1.06 mmol) was added to the reaction mixture causing a yellowcolour to appear. After 2 h, the reaction mixture was concentrated under reduced pressure.water (15 mL) was added and stirred vigorously overnight. The suspension was spun downon a centrifuge and the water decanted from the solid. The solid was returned to the reactionflask with MeOH and concentrated under reduced pressure to afford FMOC-Phe(4-Cl)-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr](1.70 g) as a crude, off-white solid.UPLC-MS (Method C ) m / z = 986.0 [M+H+NH4]2+, 977.8 [M+2H]2+This crude material (1.39g) was dissolved in DMF (30ml) treated with piperidine (10 mL)and the resulting mixture was stirred vigorously. After 1 h, the reaction mixture wasconcentrated under reduced pressure. The crude material was purified directly by columnchromatography over C18 (140 g cartridge, using DMF to load) eluting with a gradient ofMeCN (0.1% NH3) (45% to 100%; v / v, 10 CV) in water (0.1% NH3). Fractions were combinedwith the aid of MeOH / ethanol and concentrated under reduced pressure to afford the titlecompound (714 mg) as an off-white solid. Nmr shows the presence of dibenzofulvene- related byproduct.UPLC-MS (Method C): m / z = 864.6 [M-2H]2-, 887.4 [M-H+HCOO]2-, 866.4 [M+2H]2+; CEN-C-P3772PCT1H NMR (400 MHz, DMSO-D6) δ 8.65 (s, 1H), 8.48 (s, 1H), 8.25 – 7.97 (m, 4H), 7.87 (ddt, J= 16.8, 7.5, 1.0 Hz, 5H), 7.49 (s, 1H), 7.42 (td, J = 7.4, 1.2 Hz, 2H), 7.38 – 7.27 (m, 4H), 7.27– 7.14 (m, 8H), 7.11 (s, 1H), 6.98 (s, 1H), 6.85 (s, 1H), 6.77 (d, J = 5.5 Hz, 1H), 6.62 (d, J =20.9 Hz, 3H), 5.08 (d, J = 4.6 Hz, 1H), 4.92 (d, J = 5.5 Hz, 1H), 4.50 – 4.05 (m, 10H), 3.92(dt, J = 23.4, 10.7 Hz, 3H), 3.24 – 2.84 (m, 13H), 2.79 (s, 2H), 2.61 (dd, J = 13.5, 8.5 Hz,1H), 1.96 – 1.46 (m, 14H), 1.46 – 1.30 (m, 45H), 1.25 (d, J = 16.8 Hz, 1H), 1.10 – 0.96 (m,9H), 0.91 (s, 1H), 0.68 (dd, J = 16.4, 6.4 Hz, 6H). Intermediate C H-Phe(4-CF3)-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)- Dab(Boc)-Thr] (Step 1) Cbz-Phe(4-CF3)-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] H-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] (Intermediate A, Step 6) (18.50 g, 11.9 mmol) and (2S)-2-(benzyloxycarbonylamino)-3-[4- (trifluoromethyl)phenyl]propanoic acid (1.25 eq, 5.48 g, 14.9 mmol) were dissolved in DMF (0.95L).N,N-Diisopropylethylamine (6.00 eq, 12 mL, 71.6 mmol) was added followed by (1.40 eq, 6.35 g, 16.7 mmol) and the yellow solution stirred overnight. The reaction mixture was concentrated in vacuo to a brown / orange liquid. With stirring, water (100 mL) was slowly added to precipitate a white solid, which was stirred for 10 minutes before adding further water (400 mL). The precipitate was collected by filtration and dried in vacuo. The crude material was dissolved in MeOH (ca.60 degC, 900 mL), then allowed to cool to ambient temperature. A solid was precipitated by slow addition of water (700 mL). The solid was collected and air-dried. The filter cake was redissolved in MeOH (ca.60 degC, 1 L) and allowed to cool to ambient temperature. A solid was precipitated with water (500 mL) and collected to afford a cream solid. The liquor was treated with water (200-300 mL) to precipitate more solid which was collected and air-dried to afford a cream solid. The two CEN-C-P3772PCT solid batches were combined and air-dried to afford the desired product (10.00 g, 5.27 mmol, 44 % yield).UPLC-MS (Method C): m / z = 850.5 [M+H]+,1H NMR (400 MHz, DMSO-D6) δ 8.64 (s, 1H), 8.49 (s, 1H), 8.29 (d, J = 7.8 Hz, 1H), 8.21 (s,1H), 8.12 – 7.87 (m, 3H), 7.70 – 7.42 (m, 6H), 7.35 – 7.07 (m, 12H), 6.98 (s, 1H), 6.84 (s,2H), 6.68 – 6.54 (m, 3H), 5.09 (s, 1H), 4.93 (s, 3H), 4.49 – 4.15 (m, 7H), 3.98 (s, 1H), 3.93 –3.81 (m, 2H), 3.13 – 2.93 (m, 9H), 2.89 (s, 2H), 2.82 (d, J = 13.9 Hz, 3H), 1.73 (d, J = 70.0Hz, 8H), 1.52 (d, J = 9.6 Hz, 2H), 1.45 – 1.29 (m, 45H), 1.24 (s, 1H), 1.03 (d, J = 6.5 Hz, 6H),0.89 (s, 1H), 0.68 (dd, J = 16.3, 6.4 Hz, 6H).(Step 2) H-Phe(4-CF3)-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] Palladium on activated Carbon (10%) (2.55 eq, 200 mg, 1.88 mmol) was slurried with water (5mL) into a suspension of Cbz-Phe(4-CF3)-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)- DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] (1.00 eq, 2.00 g, 0.737 mmol) in ethanol (50mL) / DMF (10mL) / HCl (1.50 eq, 1.1 mL, 1.11 mmol) and the resulting mixture was stirred vigorously. The atmosphere was exchanged for hydrogen gas by backfilling from a balloon. After 19 h, UPLC-MS indicated the reaction had gone to completion. The pH was tested and found to be between 4-6. NaHCO3 (sat. sol., 5.5 mL) to adjust the pH to 9. The reaction mixture was filtered through celite eluting with ethanol (500 mL) and DMF (50mL). The resulting solution was concentrated under reduced pressure to give the title compound (1.20 g, 0.585 mmol, 79 % yield) as an off-white solid. UPLC-MS (Method C): rt = 1.39 min, m / z = 881.5 [M-2H]2-, 883.4 [M+2H]2+1H NMR (400 MHz, DMSO-D6) δ 8.64 (s, 1H), 8.49 (s, 1H), 8.29 (d, J = 7.8 Hz, 1H), 8.21 (s,1H), 8.12 – 7.87 (m, 3H), 7.70 – 7.42 (m, 6H), 7.35 – 7.07 (m, 12H), 6.98 (s, 1H), 6.84 (s,2H), 6.68 – 6.54 (m, 3H), 5.09 (s, 1H), 4.93 (s, 3H), 4.49 – 4.15 (m, 7H), 3.98 (s, 1H), 3.93 –3.81 (m, 2H), 3.13 – 2.93 (m, 9H), 2.89 (s, 2H), 2.82 (d, J = 13.9 Hz, 3H), 1.73 (d, J = 70.0Hz, 8H), 1.52 (d, J = 9.6 Hz, 2H), 1.45 – 1.29 (m, 45H), 1.24 (s, 1H), 1.03 (d, J = 6.5 Hz, 6H),0.89 (s, 1H), 0.68 (dd, J = 16.3, 6.4 Hz, 6H). Intermediate D H-[2-Octyl-L-gly]-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)- Dab(Boc)-Thr] CEN-C-P3772PCT Intermediate D may be prepared as described in WO 2020 / 201743 Preparation 4 (Boc-scaffold 4). Preparation 1 3-[2-[2-[2-(2-Aminoethoxy)ethoxy]ethoxy]ethoxy]-N-[2-[2-[(2R,3R,4R,5R,6S)-3,4,5- trihydroxy-6-methyl-tetrahydropyran-2-yl]oxyethoxy]ethyl]propanamide (Step 1): Benzyl N-[2-[2-[2-[2-[3-oxo-3-[2-[2-[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyl-tetrahydropyran-2- yl]oxyethoxy]ethylamino]propoxy]ethoxy]ethoxy]ethoxy]ethyl]carbamate (2R,3R,4R,5R,6S)-2-[2-(2-aminoethoxy)ethoxy]-6-methyl-tetrahydropyran-3,4,5-triol (WO2020074909, 2.94 g, 10.3 mmol) was dissolved in DCM (200 mL) / DMF (50 mL) , treated with 3-[2-[2-[2-[2-(benzyloxycarbonylamino)ethoxy]ethoxy]ethoxy]ethoxy]propanoic acid (4.20 g, 10.3 mmol), and N,N-Diisopropylethylamine (DIPEA) (7.2 mL, 41.2 mmol), and the resulting mixture stirred for 10 min. The solution was clear at this time. O-(7- Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (4.7 g, 12.4 mmol) was added to the reaction mixture causing a yellow colour to appear. After 19 h CEN-C-P3772PCT the reaction mixture was concentrated with the aid of toluene until ca.7 mL remained. The crude material was purified by column chromatography over C18 (400 g cartridge, using DMF to load) eluting with a gradient of MeCN (0.1% NH3) (5% to 60%; v / v) in water (0.1% NH3). Fractions were combined with the aid of ethanol and concentrated under reduced pressure to the title product (2.66 g, 4.20 mmol, 40.80% yield) as an orange oil.UPLC-MS (method B, basic) m / z = 633.4 [M+H]+1H NMR (400 MHz, DMSO-D6) δ 7.84 (t, J = 5.6 Hz, 1H), 7.38 – 7.16 (m, 6H), 4.97 (s, 2H),4.52 (d, J = 1.7 Hz, 1H), 3.62 – 3.50 (m, 4H), 3.50 – 3.41 (m, 16H), 3.37 (td, J = 5.9, 2.5 Hz,7H), 3.20 – 3.07 (m, 6H), 2.27 (t, J = 6.5 Hz, 2H), 1.09 (d, J = 6.2 Hz, 3H).(Step 2): 3-[2-[2-[2-(2-Aminoethoxy)ethoxy]ethoxy]ethoxy]-N-[2-[2-[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyl-tetrahydropyran-2- yl]oxyethoxy]ethyl]propanamide Benzyl N-[2-[2-[2-[2-[3-oxo-3-[2-[2-[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyl- tetrahydropyran-2-yl]oxyethoxy]ethylamino]propoxy]ethoxy]ethoxy]ethoxy]ethyl]carbamate (2.66 g, 4.20 mmol) and Palladium on activated carbon (314 mg, 2.95 mmol) were dissolved / suspended in EtOH (50 mL) and water (ca.10 mL) and stirred vigorously for 5 min. The flask was evacuated and backfilled with hydrogen. After 5 h, the reaction mixture was filtered through celite washing with EtOH (ca.200 mL) and concentrated under reduced pressure to give the title product (1.97 g, 3.95 mmol, 94% yield) as a colourless oil.UPLC-MS (Method B basic) m / z = 497.3 [M-H]-, 543.3 [M+HCOO]-, 499.3 [M+H]+1H NMR (400 MHz, DMSO-D6) δ 7.90 (t, J = 5.6 Hz, 1H), 4.56 (d, J = 1.6 Hz, 1H), 3.67 –3.57 (m, 5H), 3.57 (s, 1H), 3.54 – 3.44 (m, 19H), 3.40 (td, J = 6.0, 2.4 Hz, 10H), 3.22 – 3.16(m, 4H), 2.70 (t, J = 5.7 Hz, 2H), 2.31 (t, J = 6.5 Hz, 2H), 1.13 (d, J = 6.2 Hz, 3H).Intermediate 1 1-(3-(tert-Butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19- CEN-C-P3772PCT diazatetracosyl)oxy)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29- oic acid (Step 1): Benzyl 3-(tert-butyl)-5-hydroxybenzoate Benzyl bromide (1.3 mL, 10.8 mmol) was added dropwise over 5 min to asolution / suspension of 3-tert-butyl-5-hydroxybenzoic acid (1.00 eq, 2.09 g, 10.8mmol) and potassium carbonate 325 mesh (1.00 eq, 1.49 g, 10.8 mmol) in DMF (24mL) andthe resulting mixture was stirred overnight. After 20 h , the reaction mixture was combinedwith a second reaction mixture which had been performed on 11.2 mmol scale under the same conditions, and the resulting mixture was concentrated under reduced pressure to remove the DMF. The resulting residue was purified by column chromatography over silica (120 g cartridge, loaded with dichloromethane) eluting with a gradient of EtOAc (0-50, 10 CV; v / v) in iso-hexane. Fractions were combined with the aid of EtOAc and concentratedunder reduced pressure to afford benzyl 3-tert-butyl-5-hydroxy-benzoate (5.35 g,18.8 mmol,86% yield). UPLC-MS analysis (Method C): m / z = 283 [M-H]-, 285 [M+H]+1H NMR (400 MHz, DMSO-D6): δ 9.69 (s, 1H), 7.48 – 7.32 (m, 6H), 7.21 (dd, J = 2.3, 1.5Hz, 1H), 7.06 (dd, J = 2.4, 1.8 Hz, 1H), 5.32 (s, 2H), 1.26 (s, 9H). (Step 2): Benzyl 3-(2-(tert-butoxy)-2-oxoethoxy)-5-(tert-butyl)benzoate Potassium carbonate 325 mesh (287 mg, 2.08 mmol), benzyl 3-tert-butyl-5-hydroxy- benzoate (492 mg, 1.73 mmol), and tert-butylbromoacetate (0.31 mL, 2.08 mmol) were dissolved in DMF (6 mL) and stirred at room temperature for 5 min. The reaction mixture was then heated at 50 °C. After 20 h, the reaction mixture was cooled and then concentrated CEN-C-P3772PCT under reduced pressure. The residue was taken up in EtOAc (30 mL) and washed with brine (3 x 40 mL). The aqueous was re-extracted with EtOAc (3 x 30 mL), and the combined organics dried over anhydrous sodium sulphate, filtered, and concentrated under reduced pressure. The crude material was purified by column chromatography over silica (12 g cartridge, loaded with dichloromethane) eluting with a gradient of EtOAc (0% to 20%; v / v) in iso-hexane. Column fractions were combined with EtOAc to give benzyl 3-(2-tert-butoxy-2- oxo-ethoxy)-5-tert-butyl-benzoate (506 mg, 1.24 mmol, 72% yield) as a colourless oil.UPLC-MS (method C): m / z = 416 [M+NH4]+, 440 [M+H+MeCN]+, 397 [M-H]-1H NMR (400 MHz, DMSO-D6) δ 7.62 (t, J = 1.5 Hz, 1H), 7.48 – 7.31 (m, 6H), 7.21 (dq, J =2.5, 1.5 Hz, 2H), 5.35 (s, 2H), 4.73 (s, 2H), 1.38 (s, 9H), 1.28 (s, 9H).(Step 3): 2-(3-((Benzyloxy)carbonyl)-5-(tert-butyl)phenoxy)acetic acid Trifluoroacetic acid (1.2 mL, 15.7 mmol) was added to a solution of benzyl 3-(2-tert-butoxy-2- oxo-ethoxy)-5-tert-butyl-benzoate (506 mg, 1.27 mmol) in CH2Cl2 (5 mL). After 22 h, TFA (0.5 mL) was added. After a further 1 h, the reaction mixture was concentrated under reduced pressure, using toluene to azeotrope to afford 2-(3-benzyloxycarbonyl-5-tert-butyl- phenoxy)acetic acid (434 mg, 1.13 mmol, 88.84% yield) as an oil with a slight pinkish hue that solidified upon standing.UPLC-MS (method C): m / z = 341 [M-H]-, 343 [M+H]+, 360 [M+NH4]+1H NMR (400 MHz, DMSO-D6) δ 7.61 (t, J = 1.5 Hz, 1H), 7.49 – 7.32 (m, 5H), 7.28 – 7.21(m, 3H), 7.20 – 7.11 (m, 1H), 5.36 (s, 2H), 4.76 (s, 2H), 1.28 (s, 9H)(Step 4): Benzyl 3-(tert-butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19- diazatetracosyl)oxy)benzoate CEN-C-P3772PCT 3-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy]-N-[2-[2-[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy- 6-methyl-tetrahydropyran-2-yl]oxyethoxy]ethyl]propanamide (Preparation 1) (280 mg, 0.562mmol), 2-(3-((benzyloxy)carbonyl)-5-(tert-butyl)phenoxy)acetic acid (175 mg, 0.511 mmol)and triethylamine (0.28 mL, 2.04 mmol) were dissolved in DMF (20 mL), and the resulting mixture stirred for 5 min. The solution was clear at this time. O-(7-Azabenzotriazol-1-yl)- N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (233 mg, 0.613 mmol) was added to the reaction mixture causing a yellow colour to appear. After 19 h, the reaction mixture was concentrated under reduced pressure. The crude material was purified directly by column chromatography over C18 (45 g cartridge, using DMF to load) eluting with a gradient of MeCN (0.1% NH3) (35% to 75%; v / v) in water (0.1% NH3). Fractions were combined with the aid of EtOH and concentrated under reduced pressure to give the title compound (356 mg, 0.424 mmol, 83% yield) as a colourless oil.UPLC-MS (Method C) m / z = 821.5 [M-H]-, 823.5 [M+H]+, 840.6 [M+H+NH4]+1H NMR (400 MHz, DMSO-D6) δ 8.20 (t, J = 5.7 Hz, 1H), 7.90 (d, J = 5.6 Hz, 1H), 7.62 (t, J= 1.6 Hz, 1H), 7.50 – 7.33 (m, 6H), 7.28 (dd, J = 2.5, 1.8 Hz, 1H), 5.36 (s, 2H), 4.73 (t, J =4.6 Hz, 2H), 4.54 (d, J = 9.9 Hz, 4H), 3.68 – 3.37 (m, 29H), 3.28 (d, J = 5.9 Hz, 2H), 3.18 (q,J = 5.8 Hz, 3H), 1.29 (s, 9H), 1.12 (d, J = 6.2 Hz, 3H).(Step 5): 3-(tert-Butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19- diazatetracosyl)oxy)benzoic acid Benzyl 3-(tert-butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19- diazatetracosyl)oxy)benzoate (356 mg, 0.433 mmol) and palladium on activated carbon (36 mg, 0.338 mmol) were dissolved / suspended in ethanol (15 mL) / water (5 mL). The solution was then degassed and suspended under an atmosphere of hydrogen gas, and stirred vigorously. After 18 h, the reaction mixture was filtered through celite, eluting with ca.80 mL of EtOH. The filtrate was concentrated under reduced pressure to give the title product (305 mg, 0.416 mmol, 96% yield) as a colourless oil.UPLC-MS (method B, acidic): m / z = 731.5 [M-H]-, 733.5 [M+H]+, 750.5 [M+NH4]+. CEN-C-P3772PCT(Step 6): Methyl 1-(3-(tert-butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19- diazatetracosyl)oxy)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29- oate Methyl 3-[2-[2-[2-[2-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy]- ethoxy]ethoxy]ethoxy]ethoxy]propanoate hydrochloride (WO2020201743)(284 mg, 0.624 mmol) was added to a solution of 3-(tert-butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19-diazatetracosyl)oxy)benzoic acid (1.00 eq, 305 mg, 0.416 mmol) and N,N-Diisopropylethylamine (4.00 eq, 0.29 mL, 1.66 mmol) in DMF (15mL) / DMSO (3mL) andthe resulting mixture stirred for 5 min. The solution was clear at this time. O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (2.00 eq, 317 mg, 0.832 mmol) was added to the reaction mixture causing a yellow colour toappear. After 2 h the reaction mixture was concentrated under reduced pressure. The crudematerial was purified directly by column chromatography over C18 (45 g cartridge, using DMF to load) eluting with a gradient of MeCN (0.1% NH3) (25% to 60%; v / v) in water (0.1%NH3). Fractions were combined with the aid of EtOH and concentrated under reducedpressure to give the title product (453 mg,0.387 mmol, 93 % yield) as a colourless oil.UPLC-MS (Method C) m / z = 1214.8 [M+HCOO]-, 1168.8 [M-H]-, 594.6 [M+H+NH4]2+(Step 7): 1-(3-(tert-Butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19- diazatetracosyl)oxy)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29- oic acid CEN-C-P3772PCTTriethylamine (60.0 eq, 3.8 mL, 27.5 mmol) was added to a solution of methyl 1-(3-(tert-butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran- 2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19-diazatetracosyl)oxy)phenyl)-1-oxo- 5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oate (423 mg, 0.458 mmol) in water (15mL) and the resulting mixture was stirred vigorously. After 17 h, the reaction mixture wasconcentrated under reduced pressure to afford the title product (467 mg,0.437 mmol, 95 %yield) as a colourless oil. UPLC-MS (Method C) m / z = 907.6 [M-H]-, 909.6 [M+H]+, 926.7 [M+NH4]+1H NMR (400 MHz, DMSO-D6) δ 8.56 (s, 1H), 8.18 (s, 1H), 7.50 (dd, J = 3.7, 2.1 Hz, 1H),7.26 (s, 1H), 7.13 (s, 1H), 4.75 (s, 1H), 4.54 (dd, J = 13.6, 3.0 Hz, 3H), 3.71 – 3.38 (m, 48H),3.21 – 3.13 (m, 1H), 1.29 (d, J = 4.0 Hz, 9H), 1.18 – 1.08 (m, 3H)Intermediate 2 1-(3-((2,18-Dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran- 2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19-diazatetracosyl)oxy)-5-morpholinophenyl)-1-oxo- 5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oic acid (Step 1): Benzyl 3-bromo-5-(2-(tert-butoxy)-2-oxoethoxy)benzoate To a solution of benzyl 3-bromo-5-hydroxy-benzoate (3.00 g, 9.77 mmol) in dry DMF (25 mL)was added Potassium carbonate 325 mesh (1.48 g, 10.7 mmol) . The obtained mixture wasstirred for 5 minutes before tert-butylbromoacetate (1.5 mL, 10.3 mmol) was added dropwise and the reaction was allowed to stir at room temperature overnight. The reaction mixture was partitioned between brine and ethyl acetate. The phases were separated and the organic layer was washed further with brine then was dried over anhydrous sodium sulphateand filtered. Solvent was evaporated to give the crude product as a brown oil. This oil wasthen purified by column chromatography on silica gel (40 g cartridge) using a gradient of CEN-C-P3772PCT ethyl acetate (0 to 100%, v / v) in iso-hexane to provide benzyl 3-bromo-5-(2-tert-butoxy-2- oxo-ethoxy)benzoate (3.94 g, 9.35 mmol, 96% yield) as a yellow oil.UPLC-MS (Method B, acidic): m / z 438.1 [M+NH4]+1H NMR (400 MHz, DMSO-D6) δ 7.68 (t, J = 1.5 Hz, 1H), 7.50 – 7.44 (m, 3H), 7.44 – 7.33(m, 4H), 5.35 (s, 2H), 4.80 (s, 2H), 1.39 (s, 9H).(Step 2): Benzyl 3-(2-(tert-butoxy)-2-oxoethoxy)-5-morpholinobenzoate To a solution of benzyl 3-bromo-5-(2-tert-butoxy-2-oxo-ethoxy)benzoate (1.50 g, 3.56 mmol) and morpholine (0.40 mL, 4.63 mmol) in dry toluene (25 mL) was added caesium carbonate (1740 mg, 5.34 mmol) while sparging with nitrogen folllowed by RuPhos Pd G3 (298 mg,0.356 mmol). The solution was sparged with nitrogen for a further 5 min before it wasallowed to stir in a sealed reacti-vial at 70 °C overnight. The reaction mixture was analyzed by UPLC-MS which showed that the reaction was not complete and only 5% of the expected compound was formed. Hence, further RuPhos Pd G3 (149 mg, 0.178 mmol) was added and the mixture was sparged with nitrogen then allowed to stir at 80 °C. After a further 5 hours the reaction was analyzed by UPLC-MS, and had still not reached completion. Morpholine (0.31 mL, 3.56 mmol) followed by potassium tert-butoxide (400 mg, 3.56 mmol) were added while sparging with nitrogen. Finally RuPhos Pd G3 (149 mg, 0.178 mmol) was added and the reaction mixture was sparged with nitrogen with a further 5 min before it was allowed to stir at 80 °C overnight. After this time no starting material was detected by UPLC- MS. The reaction mixture was partitioned between water and ethyl acetate. The organic phase was washed further with water then with brine, dried over anhydrous sodium sulphate and filtered. The solvent was evaporated to give the crude product as a yellow oil. This oil was then purified by column chromatography on silica gel (40 g cartridge) using a gradient of ethyl acetate (5 to 40%, v / v) in iso-hexane over 10 CV to provide benzyl 3-(2-tert-butoxy-2-oxo-ethoxy)-5-morpholino-benzoate (720 mg, 1.56 mmol, 44 % yield) as a yellow oil.UPLC-MS (Method B , acidic):, m / z 428.3 [M+H]+1H NMR (400 MHz, DMSO-D6) δ 7.46 – 7.33 (m, 5H), 7.15 (dd, J = 2.4, 1.3 Hz, 1H), 6.85(dd, J = 2.3, 1.2 Hz, 1H), 6.75 (t, J = 2.3 Hz, 1H), 5.33 (s, 2H), 4.68 (s, 2H), 3.76 – 3.69 (m, CEN-C-P3772PCT4H), 3.17 – 3.10 (m, 4H), 1.38 (s, 9H).(Intermediate 2): 1-(3-((2,18-Dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19-diazatetracosyl)oxy)- 5-morpholinophenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oic acid Benzyl 3-(2-(tert-butoxy)-2-oxoethoxy)-5-morpholinobenzoate was converted into the title product following the procedures for Intermediate 1 Steps 3-7, to afford the title compound as a yellow oil.UPLC-MS (method B, acidic): m / z = 1183.8 [M-H]-1H NMR (400 MHz, DMSO-D6) δ 8.46 (t, J = 5.6 Hz, 1H), 8.08 (t, J = 5.8 Hz, 1H), 7.90 (t, J =5.6 Hz, 1H), 7.05 (dd, J = 2.2, 1.3 Hz, 1H), 6.90 (dd, J = 2.3, 1.2 Hz, 1H), 6.67 (t, J = 2.2 Hz,1H), 4.56 (d, J = 1.6 Hz, 1H), 4.49 (s, 2H), 3.74 (t, J = 4.8 Hz, 4H), 3.64 – 3.54 (m, 8H), 3.54– 3.42 (m, 56H), 3.40 (ddd, J = 7.3, 4.7, 2.4 Hz, 9H), 3.29 (q, J = 5.9 Hz, 4H), 3.22 – 3.12(m, 8H), 2.42 (dt, J = 6.5, 1.5 Hz, 2H), 2.31 (t, J = 6.5 Hz, 2H), 1.12 (d, J = 6.2 Hz, 3H). Intermediate 3 1-(3-((2,18-Dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran- 2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19-diazatetracosyl)oxy)-5-(trifluoromethyl)phenyl)-1- oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oic acid (Step 1): Benzyl 3-hydroxy-5-(trifluoromethyl)benzoate To a solution of 3-hydroxy-5-(trifluoromethyl)benzoic acid (3.00 g, 14.6 mmol) wasadded potassium carbonate 325 mesh (1.00 eq, 2011 mg, 14.6 mmol) . The obtainedmixture was stirred for 5 minutes before benzyl bromide (1.00 eq, 1.7 mL, 14.6 mmol) wasadded dropwise and the reaction was allowed to stir at room temperature for 5 hours.The reaction mixture was partitioned between brine and ethyl acetate. The phases wereseparated and the organic phase was washed twice with brine, dried over anhydrous sodium CEN-C-P3772PCT sulphate and filtered. The solvent was evaporated to give the crude product as a brown oil, which was purified by column chromatography on silica gel (80 g cartridge) using a gradientof ethyl acetate (0 to 60%, v / v) in iso-hexane to provide benzyl 3-hydroxy-5-(trifluoromethyl)benzoate (3.31 g,11.2 mmol, 77 % yield) as a yellowish solid.UPLC-MS (method B, acidic): m / z 295.0 [M-H]-1H NMR (400 MHz, DMSO-D6) δ 10.64 (s, 1H), 7.63 (d, J = 2.1 Hz, 2H), 7.50 – 7.45 (m, 2H),7.45 – 7.34 (m, 3H), 7.32 (t, J = 2.1 Hz, 1H), 5.36 (s, 2H).19F NMR (376 MHz, DMSO-D6) δ -61.47.(Intermediate 3): 1-(3-((2,18-Dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19-diazatetracosyl)oxy)- 5-(trifluoromethyl)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oic acid Benzyl 3-hydroxy-5-(trifluoromethyl)benzoate was converted into the title compound following the procedure of Intermediate 1, steps 2-7, to afford the title product as a pale yellow oil.UPLC-MS (Method B, acidic): m / z = 1166.7 [M-H]-1H NMR (400 MHz, DMSO-D6) δ 8.83 (t, J = 5.5 Hz, 1H), 8.25 (t, J = 5.7 Hz, 1H), 7.90 (t, J =5.6 Hz, 1H), 7.81 (td, J = 1.5, 0.7 Hz, 1H), 7.74 (t, J = 2.0 Hz, 1H), 7.45 (t, J = 2.1 Hz, 1H),4.66 (s, 3H), 4.56 (d, J = 1.7 Hz, 1H), 3.71 – 3.24 (m, 71H), 3.22 – 3.13 (m, 3H), 2.42 (t, J =6.4 Hz, 2H), 2.31 (t, J = 6.5 Hz, 2H), 1.12 (d, J = 6.2 Hz, 3H).19F NMR (376 MHz, DMSO-D6) δ -61.07Intermediate 4 1-(3-(tert-Butyl)-5-(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)ethoxy)phenyl)-1-oxo- 5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oic acid CEN-C-P3772PCT (Step 1): Benzyl 3-[2-oxo-2-[2-[2-[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyl-tetrahydro-pyran-2-yl]oxyethoxy]ethylamino]ethoxy]-5-phenyl-benzoate 2-(3-((benzyloxy)carbonyl)-5-(tert-butyl)phenoxy)acetic acid (Intermediate 1, step 3, 260 mg, 0.683 mmol), (2R,3R,4R,5R,6S)-2-[2-(2-aminoethoxy)ethoxy]-6-methyl-tetrahydropyran-3,4,5-triol (WO 2020 / 074909, 237 mg, 0.943 mmol), and N,N-Diisopropylethylamine (DIPEA)(0.60 mL, 3.42 mmol) were dissolved in DMF (30 mL) and the resulting mixture stirred for 5 min. The solution was clear at this time. O-(7-Azabenzotriazol-1-yl)-N,N,N',N'- tetramethyluronium hexafluorophosphate (HATU) (338 mg, 0.888 mmol) was added to the reaction mixture causing a yellow colour to appear. After 19 h, the reaction mixture was concentrated under reduced pressure. The crude material was purified directly by column chromatography over C18 (45 g cartridge, using DMF to load) eluting with a gradient of MeCN (0.1% NH3) (40% to 70%; v / v) in water (0.1% NH3). Fractions were combined with the aid of EtOH and concentrated under reduced pressure to give the title compound (265 mg, 0.433 mmol, 63% yield) as an off-white oily foam.UPLC-MS (Method C): m / z = 574.3 [M-H]-, 576.4 [M+H]+, 593.4 [M+NH4]+1H NMR (400 MHz, DMSO-D6) δ 8.22 (t, J = 5.8 Hz, 1H), 7.62 (t, J = 1.5 Hz, 1H), 7.48 –7.33 (m, 6H), 7.28 (dd, J = 2.5, 1.7 Hz, 1H), 5.36 (s, 2H), 4.75 – 4.70 (m, 2H), 4.58 – 4.51(m, 4H), 3.59 (ddd, J = 7.8, 5.7, 3.4 Hz, 2H), 3.52 – 3.35 (m, 8H), 3.32 – 3.25 (m, 2H), 3.17(td, J = 9.4, 5.5 Hz, 1H), 1.29 (s, 9H), 1.12 (d, J = 6.2 Hz, 3H). (Intermediate 4): 1-(3-(tert-Butyl)-5-(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy- 6-methyltetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)ethoxy)phenyl)-1-oxo- 5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oic acid Benzyl 3-[2-oxo-2-[2-[2-[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyl-tetrahydro-pyran-2- yl]oxyethoxy]ethylamino]ethoxy]-5-phenyl-benzoate was converted into the title compound using the conditions described for Intermediate 1 steps 5-7 to afford the title compound as a colourless oil. CEN-C-P3772PCTUPLC-MS (Method C, basic): m / z = 907.6 [M-H]-, 909.6 [M+H]+, 926.7 [M+NH4]+1H NMR (400 MHz, DMSO-D6) δ 8.56 (s, 1H), 8.18 (s, 1H), 7.50 (dd, J = 3.7, 2.1 Hz, 1H), 7.26(s, 1H), 7.13 (s, 1H), 4.75 (s, 1H), 4.54 (dd, J = 13.6, 3.0 Hz, 3H), 3.71 – 3.38 (m, 48H), 3.21– 3.13 (m, 1H), 1.29 (d, J = 4.0 Hz, 9H), 1.18 – 1.08 (m, 3H)Intermediate 5 1-(3-(2-((3-(((2R,3R,4R,5S,6R)-3-Acetamido-5-(((2S,3R,4S,5S,6R)-3,5-dihydroxy-6- (hydroxymethyl)-4-(((2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro- 2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4-hydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)amino)-2-oxoethoxy)-5-(tert- butyl)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oic acid (Step 1): Benzyl 3-(2-((3-(((2R,3R,4R,5S,6R)-3-acetamido-5-(((2S,3R,4S,5S,6R)-3,5-dihydroxy-6-(hydroxymethyl)-4-(((2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4- hydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)amino)-2-oxoethoxy)- 5-(tert-butyl)benzoate 2-(3-((Benzyloxy)carbonyl)-5-(tert-butyl)phenoxy)acetic acid (1.55 g, 4.53 mmol) was addedto a solution of 3-(((2R,3R,4R,5S,6R)-3-acetamido-5-(((2S,3R,4S,5S,6R)-3,5-dihydroxy-6-(hydroxymethyl)-4-(((2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H- pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4-hydroxy-6-(hydroxymethyl)tetrahydro-2H- pyran-2-yl)oxy)propyl)amine hydrochloride (which can be prepared according to the methodsdescribed by Bovin et al (Mendeleev Communications (2002), (4), 143-145), 3.75 g, 5.87mmol) and triethylamine (4.00 eq, 2.5 mL, 18.1 mmol) in DMF (160mL) / DMSO (40mL) andthe resulting mixture stirred for 15 min. The solution was clear at this time. O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (1.30 eq, 2.24 g, 5.89 mmol) was added to the reaction mixture causing a yellow colour to CEN-C-P3772PCTappear. After 2 h , the reaction mixture was concentrated under reduced pressure to aconstant volume (residual DMSO). The crude material was purified directly by column chromatography over C18 (375 g cartridge, using DMSO / DMF to load) eluting with a gradientof MeCN (0.1% NH3) (25% to 60%; v / v, 10 CV) in water (0.1% NH3). Fractions were combinedwith the aid of EtOH and concentrated under reduced pressure to give the title product (4.20g,4.21 mmol, 93 % yield) as a white solid.UPLC-MS (Method C): m / z = 925.4 [M-H]-, 927.5 [M+H]+1H NMR (400 MHz, DMSO-D6) δ 8.15 (t, J = 5.8 Hz, 1H), 7.75 (d, J = 7.9 Hz, 1H), 7.62 (t, J= 1.6 Hz, 1H), 7.51 – 7.31 (m, 6H), 7.28 (dd, J = 2.5, 1.7 Hz, 1H), 5.36 (s, 2H), 5.11 (d, J = 5.1Hz, 1H), 4.82 (d, J = 3.5 Hz, 1H), 4.76 – 4.66 (m, 2H), 4.63 (d, J = 4.4 Hz, 3H), 4.54 (s, 2H),4.47 (t, J = 5.4 Hz, 1H), 4.40 – 4.32 (m, 4H), 4.28 (d, J = 7.4 Hz, 2H), 3.99 (t, J = 6.6 Hz, 1H),3.84 (s, 1H), 3.73 (d, J = 9.5 Hz, 3H), 3.66 – 3.37 (m, 18H), 3.21 – 3.08 (m, 2H), 1.79 (s, 3H),1.67 – 1.57 (m, 2H), 1.29 (s, 9H)(Step 2): 3-(2-((3-(((2R,3R,4R,5S,6R)-3-Acetamido-5-(((2S,3R,4S,5S,6R)-3,5-dihydroxy-6-(hydroxymethyl)-4-(((2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4- hydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)amino)-2-oxoethoxy)- 5-(tert-butyl)benzoic acid Palladium on activated carbon (10%) (0.937 eq, 420 mg, 3.95 mmol) was slurried with Water(50mL) into a solution of the product from step 1 (4.20 g, 4.21 mmol) in Ethanol (200mL) andthe resulting mixture was stirred vigorously (NB The solid was not very soluble and so the flask was mechanically agitated several times to rinse solid into solution). The atmospherewas exchanged for hydrogen gas by backfilling from a balloon . After 90 min, another portionof EtOH / water was added (40 mL) to aid solubility.After 18 h, the reaction mixture was filtered through celite eluting with: EtOH (ca. 800 mL),water (ca.50 mL) then DMF (150 mL). The resulting solution was concentrated under reducedpressure to give the title compound (4.00 g,4.06 mmol, 96 % yield) as a white solid. CEN-C-P3772PCTUPLC-MS (Method C): m / z = 835.4 [M-H]-, 837.4 [M+H]+1H NMR (400 MHz, DMSO-D6) δ 8.11 (s, 1H), 7.95 (s, 2H), 7.78 (s, 1H), 7.58 (t, J = 1.6 Hz,1H), 7.28 (s, 1H), 7.20 (s, 1H), 5.12 (s, 1H), 4.82 (d, J = 3.6 Hz, 1H), 4.74 (s, 1H), 4.71 – 4.58(m, 3H), 4.52 (s, 2H), 4.48 (s, 1H), 4.35 (d, J = 4.3 Hz, 2H), 4.28 (d, J = 7.5 Hz, 2H), 3.99 (t, J= 6.6 Hz, 1H), 3.84 (s, 1H), 3.76 (d, J = 18.8 Hz, 3H), 3.66 – 3.36 (m, 12H), 3.26 (d, J = 6.9Hz, 2H), 3.16 (dt, J = 21.5, 6.8 Hz, 2H), 1.80 (s, 3H), 1.71 – 1.57 (m, 2H), 1.29 (s, 9H)(Step 3): Methyl 1-(3-(2-((3-(((2R,3R,4R,5S,6R)-3-acetamido-5-(((2S,3R,4S,5S,6R)-3,5-dihydroxy-6-(hydroxymethyl)-4-(((2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4- hydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)amino)-2-oxoethoxy)- 5-(tert-butyl)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oate A solution of methyl 3-[2-[2-[2-[2-[2-[2-[2-(2-aminoethoxy)ethoxy]ethoxy]ethoxy]- ethoxy]ethoxy]ethoxy]ethoxy]propanoate hydrochloride (WO2020201743, 3.26 g, 7.16 mmol) in DMF (150mL) / DMSO (30mL) was added to a solution of 3-(2-((3-(((2R,3R,4R,5S,6R)-3- acetamido-5-(((2S,3R,4S,5S,6R)-3,5-dihydroxy-6-(hydroxymethyl)-4-(((2R,3R,4S,5R,6R)- 3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2- yl)oxy)-4-hydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)amino)-2- oxoethoxy)-5-(tert-butyl)benzoic acid (1.00 eq, 4.00 g, 4.06 mmol) and triethylamine (5.12 eq, 2.9 mL, 20.8 mmol) , and the resulting mixture stirred for 15 min. The solution was clear at this time. HATU (2.36 eq, 3.64 g, 9.57 mmol) was added to the reaction mixture causing a yellow colour to appear. After 18 h, the reaction mixture was concentrated under reduced pressure (including some of the DMSO). The crude material was purified directly by column chromatography over C18 (375 g cartridge, using DMSO / water to load) eluting with a gradient of MeCN (0.1% NH3) (5% to 50%; v / v) in water (0.1% NH3). Fractions were combined with the aid of iPrOH and concentrated under reduced pressure to give the title product (4.20 g,3.21 mmol, 79 % yield) as an oily off-white foam.UPLC-MS (Method C): m / z = 646.7 [M+H+NH4]2+ CEN-C-P3772PCT1H NMR (400 MHz, DMSO-D6) δ 8.53 (t, J = 5.6 Hz, 1H), 8.09 (t, J = 5.8 Hz, 1H), 7.74 (d, J =8.1 Hz, 1H), 7.50 (t, J = 1.5 Hz, 1H), 7.25 (dd, J = 2.4, 1.4 Hz, 1H), 7.13 (dd, J = 2.4, 1.7 Hz, 1H), 5.09 (d, J = 5.2 Hz, 1H), 4.82 (d, J = 3.5 Hz, 1H), 4.72 (t, J = 5.0 Hz, 1H), 4.68 (d, J = 7.6Hz, 1H), 4.64 – 4.59 (m, 3H), 4.50 (s, 2H), 4.45 (t, J = 5.4 Hz, 1H), 4.38 – 4.32 (m, 2H), 4.29(t, J = 7.7 Hz, 2H), 3.99 (t, J = 6.5 Hz, 1H), 3.84 (s, 1H), 3.76 (dddd, J = 14.3, 6.5, 4.3, 2.1 Hz,3H), 3.68 – 3.36 (m, 52H), 3.29 – 3.23 (m, 1H), 3.17 (tq, J = 13.2, 6.4 Hz, 2H), 1.80 (s, 3H),1.65 (t, J = 6.9 Hz, 2H), 1.29 (s, 9H)(Step 4): 1-(3-(2-((3-(((2R,3R,4R,5S,6R)-3-Acetamido-5-(((2S,3R,4S,5S,6R)-3,5-dihydroxy-6-(hydroxymethyl)-4-(((2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4- hydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)amino)-2-oxoethoxy)- 5-(tert-butyl)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oic acid Triethylamine (60.0 eq, 28 mL, 198 mmol) was added to a solution of the product from Step 3 (1.00 eq, 4.20 g, 3.30 mmol) in Water (220mL) and the resulting mixture was stirredvigorously. After 1 h, the reaction mixture was concentrated under reduced pressure toafford (V) (4.30 g,3.21 mmol, 97 % yield) as an off-white oily foam.UPLC-MS (Method C): m / z = 629.0 [M-2H]2-, 639.8 [M+H+NH4]2+1H NMR (400 MHz, DMSO-D6) δ 8.55 (t, J = 5.6 Hz, 1H), 8.13 (t, J = 5.8 Hz, 1H), 7.77 (d, J =7.9 Hz, 1H), 7.50 (t, J = 1.5 Hz, 1H), 7.26 (dd, J = 2.5, 1.4 Hz, 1H), 7.13 (dd, J = 2.4, 1.7 Hz, 1H), 5.11 (s, 1H), 4.82 (d, J = 3.5 Hz, 1H), 4.63 (s, 3H), 4.51 (s, 2H), 4.37 (s, 2H), 4.29 (dd, J= 7.8, 6.2 Hz, 2H), 3.99 (t, J = 6.6 Hz, 1H), 3.84 (s, 1H), 3.81 – 3.69 (m, 3H), 3.68 – 3.37 (m,51H), 3.28 (s, 2H), 3.17 (dp, J = 19.2, 6.5 Hz, 3H), 2.41 (t, J = 6.4 Hz, 2H), 1.80 (s, 3H), 1.65 (p, J = 6.9 Hz, 2H), 1.29 (s, 9H) CEN-C-P3772PCT diazatetracosyl)oxy)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29- oyl-[L-(2-octyl)-Gly]-Dab-Thr-Dap-cyclo-[Dab-Dab-DPhe-Leu-Dab-Dab-Thr](Step 1): 1-(3-(tert-Butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19- diazatetracosyl)oxy)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29- oyl-[L-(2-Octyl)-Gly]-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu- Dab(Boc)-Dab(Boc)-Thr] H-[L-(2-octyl)-Gly]-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)-DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] (Intermediate D, 90 mg, 0.0524 mmol), 1-(3-(tert-butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22- pentaoxa-3,19-diazatetracosyl)oxy)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2- azanonacosan-29-oic acid (Intermediate 1, 70 mg, 0.0576 mmol) and triethylamine (0.029 mL, 0.209 mmol) were dissolved in DMF (5 mL), and the resulting mixture stirred for 5 min. The solution was clear at this time. O-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (30 mg, 0.0785 mmol) was added to the reaction mixture causing a yellow colour to appear. After 2 h, the reaction mixture was concentrated under reduced pressure. The crude material was purified by column chromatography over C18 (23g cartridge) eluting with a gradient of MeCN (0.1% NH3) (45% to 80%; v / v) in water (0.1%NH3). Fractions were combined with the aid of EtOH and concentrated under reducedpressure to afford the desired product (99 mg, 0.0346 mmol, 66% yield) as a white solid.UPLC-MS (Method C): m / z = 951.4 [M-3H]3-, 966.7 [M-2H+HCOO]3-, 964.8 [M+H+2NH4]3+;1H NMR (400 MHz, DMSO-D6) δ 8.63 (s, 1H), 8.51 (d, J = 5.7 Hz, 1H), 8.22 – 8.06 (m, 2H),8.08 – 7.89 (m, 2H), 7.87 (d, J = 6.2 Hz, 1H), 7.78 (s, 1H), 7.50 (s, 2H), 7.24 (d, J = 4.7 Hz,4H), 7.19 (s, 1H), 7.12 (s, 2H), 6.83 (s, 1H), 6.76 (s, 1H), 6.66 – 6.54 (m, 2H), 5.03 (s, 1H),4.90 (s, 1H), 4.77 – 4.65 (m, 1H), 4.56 (s, 1H), 4.51 (d, J = 6.1 Hz, 3H), 4.43 – 4.05 (m, 3H),4.02 – 3.77 (m, 3H), 3.67 – 3.38 (m, 71H), 3.24 – 2.69 (m, 17H), 2.36 – 2.26 (m, 5H), 1.96 –1.42 (m, 11H), 1.44 – 1.32 (m, 45H), 1.32 – 1.16 (m, 28H), 1.12 (d, J = 6.2 Hz, 3H), 1.02 (t, J CEN-C-P3772PCT = 7.0 Hz, 6H), 0.85 (t, J = 6.6 Hz, 3H), 0.69 (appears as dd, J = 16.8, 6.5 Hz, 6H)(Step 2): 1-(3-(tert-Butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19- diazatetracosyl)oxy)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29- oyl-[L-(2-octyl)-Gly]-Dab-Thr-Dap-cyclo-[Dab-Dab-DPhe-Leu-Dab-Dab-Thr] Trifluoroacetic acid (2.9 mL, 37.3 mmol) was added to a solution of 1-(3-(tert-butyl)-5-((2,18- dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)- 6,9,12,15,22-pentaoxa-3,19-diazatetracosyl)oxy)phenyl)-1-oxo-5,8,11,14,17,20,23,26- octaoxa-2-azanonacosan-29-oyl-[L-octyl Gly]-Dab(Boc)-Thr-Dap(Boc)-cyclo-[Dab-Dab(Boc)- DPhe-Leu-Dab(Boc)-Dab(Boc)-Thr] (99 mg, 0.0346 mmol) in dichloromethane (4 mL) and water (2 drops). After 20 min , the reaction was quenched with MTBE (25 mL) and the reaction mixture concentrated under reduced pressure. The crude material was purified by column chromatography over C18 (23 g cartridge, loaded with water / DMF) eluting with a gradient of MeCN (0.1% TFA) (5% to 50%; v / v) in water (0.1% TFA). Fractions were combined with water and concentrated under reduced pressure. The residue was freeze- dried overnight to afford the title product (62 mg, 0.0212 mmol, 61% yield) as a fluffy white solid.UPLC-MS (Method D): m / z = 786.5 [M+3H]3+, 1179.3 [M+2H]2+1H NMR (400 MHz, METHANOL-D4) δ 7.56 (t, J = 1.5 Hz, 1H), 7.36 – 7.16 (m, 7H), 4.72 (d,J = 1.7 Hz, 1H), 4.60 (s, 2H), 4.56 – 4.25 (m, 7H), 4.23 – 4.04 (m, 4H), 3.95 – 3.85 (m, 1H),3.85 – 3.44 (m, 70H), 3.42 – 3.32 (m, 5H), 3.17 – 2.87 (m, 11H), 2.75 – 2.57 (m, 1H), 2.56 –2.39 (m, 4H), 2.31 – 1.87 (m, 8H), 1.81 – 1.74 (m, 2H), 1.58 – 1.17 (m, 36H), 0.96 – 0.85 (m,4H), 0.79 (d, J = 6.5 Hz, 3H), 0.72 (d, J = 6.4 Hz, 3H)Examples 2-7 were prepared according to the method of Example 1, with data shown inTable 1. Example 2: 1-(3-(tert-Butyl)-5-((2,18-dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19- diazatetracosyl)oxy)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29- oyl-Phe(4-F)-Dab-Thr-Dap-cyclo-[Dab-Dab-DPhe-Leu-Dab-Dab-Thr]1H NMR (400 MHz, METHANOL-D4) δ 7.56 (t, J = 1.6 Hz, 1H), 7.36 – 7.17 (m, 9H), 7.09 –6.94 (m, 2H), 4.71 (d, J = 1.7 Hz, 1H), 4.60 (s, 2H), 4.54 – 4.25 (m, 8H), 4.21 – 4.14 (m, 2H), CEN-C-P3772PCT4.12 – 4.06 (m, 1H), 3.89 – 3.43 (m, 66 H), 3.39 – 3.32 (m, 4H), 3.12 – 2.84 (m, 12H), 2.66 –2.51 (m, 1H), 2.49 – 2.36 (m, 4H), 2.27 – 2.03 (m, 6H), 2.01 – 1.86 (m, 2H), 1.58 – 1.46 (m,1H), 1.44 – 1.31 (m, 11H), 1.26 (d, J = 6.2 Hz, 3H), 1.21 (appears as dd, J = 8.3, 6.3 Hz,6H), 0.96 – 0.83 (m, 1H), 0.79 (d, J = 6.5 Hz, 3H), 0.72 (d, J = 6.4 Hz, 3H).Example 3: 1-(3-((2,18-Dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19-diazatetracosyl)oxy)- 5-morpholinophenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oyl -Phe(4-F)-Dab-Thr-Dap-cyclo-[Dab-Dab-DPhe-Leu-Dab-Dab-Thr]1H NMR (400 MHz, METHANOL-D4) δ 7.36 – 7.18 (m, 7H), 7.10 (dd, J = 2.3, 1.4 Hz, 1H),7.07 – 6.99 (m, 2H), 6.93 (dd, J = 2.2, 1.3 Hz, 1H), 6.77 (t, J = 2.2 Hz, 1H), 4.71 (d, J = 1.8Hz, 1H), 4.58 (s, 2H), 4.54 – 4.25 (m, 9H), 4.17 (dd, J = 11.1, 4.0 Hz, 1H), 4.08 (d, J = 4.9Hz, 1H), 3.86 – 3.80 (m, 6H), 3.79 – 3.73 (m, 1H), 3.73 – 3.55 (m, 61H), 3.55 – 3.45 (m, 6H),3.41 – 3.33 (m, 4H), 3.22-3.19 (m, 4H), 3.17 – 2.84 (m, 11H), 2.65 – 2.53 (m, 1H), 2.45 (t, J= 6.2 Hz, 3H), 2.32 – 2.05 (m, 6H), 1.95 (d, J = 11.2 Hz, 2H), 1.44 – 1.28 (m, 2H), 1.26 (d, J= 6.2 Hz, 3H), 1.21 (appears as dd, J = 8.2, 6.4 Hz, 6H), 1.01 – 0.83 (m, 1H), 0.82 – 0.68 (m,6H). Example 4: 1-(3-((2,18-Dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19-diazatetracosyl)oxy)- 5-(trifluoromethyl)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oyl -Phe(4-F)-Dab-Thr-Dap-cyclo-[Dab-Dab-DPhe-Leu-Dab-Dab-Thr]1H NMR (400 MHz, METHANOL-D4) δ 7.80 (s, 1H), 7.73 (s, 1H), 7.50 (s, 1H), 7.32 – 7.21(m, 9H), 4.72 (d, J = 1.7 Hz, 1H), 4.70 (s, 2H), 4.55 – 4.25 (m, 9H), 4.17 (s, 1H), 4.08 (d, J =5.0 Hz, 1H), 3.81 (dd, J = 3.5, 1.7 Hz, 2H), 3.79 – 3.74 (m, 1H), 3.74 – 3.56 (m, 61H), 3.56 –3.46 (m, 6H), 3.40 – 3.34 (m, 5H), 3.07 (appears as dt, J = 18.9, 8.6 Hz, 12H), 2.92 (dd, J =17.7, 10.1 Hz, 3H), 2.58 (s, 1H), 2.46 (t, J = 6.1 Hz, 3H), 2.31 – 2.01 (m, 6H), 1.95 (d, J =10.6 Hz, 2H), 1.57 – 1.33 (m, 2H), 1.26 (d, J = 6.2 Hz, 3H), 1.23 (d, J = 6.3 Hz, 3H), 1.21 (d,J = 6.3 Hz, 3H), 0.98 – 0.82 (m, 1H), 0.78 (d, J = 6.5 Hz, 3H), 0.72 (d, J = 6.5 Hz, 3H).Example 5: 1-(3-(tert-Butyl)-5-(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)ethoxy)phenyl)-1-oxo- 5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oyl-Phe(4-F)-Dab-Thr-Dap-cyclo- [Dab-Dab-DPhe-Leu-Dab-Dab-Thr] CEN-C-P3772PCT1H NMR (400 MHz, METHANOL-D4) δ 7.55 (t, J = 1.6 Hz, 1H), 7.33 – 7.20 (m, 9H), 7.07 –6.98 (m, 2H), 4.71 (d, J = 1.7 Hz, 1H), 4.60 (s, 2H), 4.56 – 4.26 (m, 8H), 4.23 – 4.13 (m, 2H),4.13 – 4.04 (m, 1H), 3.88 – 3.79 (m, 2H), 3.79 – 3.52 (m, 48H), 3.51 – 3.44 (m, 4H), 3.41 –3.33 (m, 3H), 3.17 – 2.86 (m, 12H), 2.65 – 2.52 (m, 1H), 2.51 – 2.35 (m, 2H), 2.30 – 2.03 (m,6H), 2.03 – 1.83 (m, 2H), 1.58 – 1.46 (m, 1H), 1.43 – 1.28 (m, 11 H), 1.25 (d, J = 6.2 Hz,3H), 1.21 (appears as dd, J = 8.4, 6.4 Hz, 6H), 0.98 – 0.82 (m, 1H), 0.78 (d, J = 6.4 Hz, 3H),0.72 (d, J = 6.4 Hz, 3H)Example 6: 1-(3-((2,18-Dioxo-24-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-6,9,12,15,22-pentaoxa-3,19-diazatetracosyl)oxy)- 5-(trifluoromethyl)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oyl -Phe(4-Cl)-Dab-Thr-Dap-cyclo-[Dab-Dab-DPhe-Leu-Dab-Dab-Thr]1H NMR (400 MHz, METHANOL-D4) δ 7.80 (s, 1H), 7.73 (s, 1H), 7.50 (s, 1H), 7.32 – 7.21(m, 9H), 4.72 (d, J = 1.7 Hz, 1H), 4.70 (s, 2H), 4.55 – 4.25 (m, 9H), 4.17 (s, 1H), 4.08 (d, J =5.0 Hz, 1H), 3.81 (dd, J = 3.5, 1.7 Hz, 2H), 3.79 – 3.74 (m, 1H), 3.74 – 3.56 (m, 60H), 3.56 –3.46 (m, 6H), 3.40 – 3.34 (m, 5H), 3.07 (appears as dt, J = 18.9, 8.6 Hz, 12H), 2.92 (dd, J =17.7, 10.1 Hz, 3H), 2.58 (s, 1H), 2.46 (t, J = 6.1 Hz, 3H), 2.31 – 2.01 (m, 6H), 1.95 (d, J =10.6 Hz, 2H), 1.57 – 1.33 (m, 2H), 1.26 (d, J = 6.2 Hz, 3H), 1.21 (appears as dd, J = 9.0, 6.3Hz, 6H), 0.96-0.82 (m, 1H), 0.78 (d, J = 6.5 Hz, 3H), 0.72 (d, J = 6.5 Hz, 3H). Example 7: 1-(3-(2-((3-(((2R,3R,4R,5S,6R)-3-Acetamido-5-(((2S,3R,4S,5S,6R)-3,5- dihydroxy-6-(hydroxymethyl)-4-(((2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6- (hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4- hydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)propyl)amino)-2-oxoethoxy)- 5-(tert-butyl)phenyl)-1-oxo-5,8,11,14,17,20,23,26-octaoxa-2-azanonacosan-29-oyl- Phe(4-CF3)-Dab-Thr-Dap-cyclo-[Dab-Dab-DPhe-Leu-Dab-Dab-Thr]1H NMR (400 MHz, METHANOL-D4) δ 7.60 (d, J = 8.1 Hz, 2H), 7.55 (t, J = 1.5 Hz, 1H), 7.46(d, J = 8.0 Hz, 2H), 7.34 – 7.19 (m, 7H), 5.04 (d, J = 2.9 Hz, 1H), 4.59 (s, 2H), 4.56 – 4.28(m, 10H), 4.26 – 4.13 (m, 3H), 4.11 – 4.05 (m, 1H), 4.05 – 4.00 (m, 1H), 3.95 – 3.49 (m,52H), 3.43 – 3.33 (m, 3H), 3.28 – 3.22 (m, 2H), 3.17 – 2.86 (m, 14H), 2.62 – 2.52 (m, 1H),2.48 – 2.36 (m, 2H), 2.29 – 2.04 (m, 6H), 2.03 – 1.88 (m, 6H), 1.84 – 1.70 (m, 2H), 1.57 –1.45 (m, 1H), 1.45 – 1.37 (m, 1H), 1.37 – 1.27 (m, 10H), 1.21 (appears as dd, J = 10.8, 6.4Hz, 6H), 0.98 – 0.86 (m, 1H), 0.78 (d, J = 6.5 Hz, 3H), 0.71 (d, J = 6.4 Hz, 3H). CEN-C-P3772PCT Table 1 ExampleF S2 R1 R2 Molecular Accurate Mass m / z UPLC RTStarting Number Formula (min) Materials Method D -(CH2)2-O- (CH2)2-NH- CO- 1179.3 [M+2H]2+1A Tert butyl octyl C109H190N20O36 2355.372 2 4 H]3+1.25 Int D / Int 1(CH CH O) 786.5 [M+3 -(CH2)2-NH- CO-CH2- -(CH2)2-O- C108H179FN20O3 1177.2 [M+2H]2+2A(CH2)2-NH-Tert butyl 4-F benzyl2351.281.14 Int A / Int 16 785.1 [M+3H]3+CO-

[0002] CEN-C-P3772PCT ExampleF S2 R1 R2 Molecular Accurate Mass m / z UPLC RTStarting Number Formula (min) Materials Method D (CH2CH2O)4-(CH2)2-NH- CO-CH2- -(CH2)2-O- (CH2)2-NH- CO- morpholi C108H178FN21O3 1191.5 [M+2H]2+3A4-F benzyl 2380.271.07 Int A / Int 2(CH2CH2O)4 nyl 7 794.8 [M+3H]3+-(CH2)2-NH- CO-CH2- -(CH2)2-O- (CH2)2-NH- CO- C105H170F4N20O 1183.1 [M+2H]2+4ACF34-F benzyl 2363.201.13 Int A / Int 3(CH2CH2O)4 36 789.0 [M+3H]3+-(CH2)2-NH- CO-CH2- -(CH2)2-O- 1053.5 [M+2H]2+5A(CH2)2-NH-Tert butyl 4-F benzyl C97H158FN19O31 2104.14 1.12 Int A / Int 4702.7 [M+3H]3+CO-CH2- -(CH2)2-O- 4-Cl C105H186ClF3N2 1190.8 [M+2H]2+6ACF32379.171.13 Int B / int 3(CH2)2-NH- benzyl 0O36 794.05 [M+3H]3+

[0003] CEN-C-P3772PCT ExampleF S2 R1 R2 Molecular Accurate Mass m / z UPLC RTStarting Number Formula (min) Materials Method D CO- (CH2CH2O)4-(CH2)2-NH- CO-CH2- –(CH2)3- 4-CF3 C111H179F3N20O 7BNH-CO- Tert butyl2505.25 836.4 [M+3H]3+ 1.11 Int C / Int 5benzyl 41 CH2-

[0004] CEN-C-P3772PCTReference Examples 1 and 2Reference Example 1 was prepared according to Example 2 of WO 2020 / 201743. Reference Example 2 was prepared according to Example 4 of WO 2020 / 201743. BIOLOGICAL ASSAYS1. Anti-Rhamnose IgG Recruitment AssayTable 2 shows the recruitment of IgG anti-rhamnose antibodies to the surface of E. coliATCC 700973. Polymyxin B which does not contain an antibody-recognition element, is used as a negative control. Plate based antibody recruitment assay using an anti-Rhamnose antibody.Fluorescence based assay was used to demonstrate binding of compounds of Examples 1-6to Escherichia coli bacteria and binding to monoclonal hIgG1 anti-Rhamnose antibodies. Afluorescently labelled anti human IgG was used to detect the anti-Rhamnose antibody binding. Method The assays were carried out in black 96 well flat-bottomed plates (ThermoScientific).Escherichia coli (ATCC 700973) was grown in RPMI 1640 (Invitrogen - 11835030),supplemented with 10% LB broth (Miller) to mid exponential phase. Subsequently, the bacteria were washed once with Hank's Balanced Salt Solution with calcium and magnesium (HBSS+ / +) and then resuspended in HBSS+ / + at a bacterial density of 8x107colony forming units (CFU) / mL.1.8x107CFU were then incubated for 60 minutes in the presence of human anti-Rhamnose monoclonal antibody (clone 023.102, Bryson, S. (2016). Journal ofImmunology 96(11), 4723–4730) at a final concentration of 100 µg / mL with Examples 1-6compounds or Polymyxin B at 0.3 to 20 µM end concentration or a vehicle only control, at 25°C and shaking at 300 rpm. The bacteria were then centrifuged, and the supernatantremoved prior to addition of 100 µL of the FITC labelled secondary antibody (Goat pAb anti-human IgG Fc – FITC (Abcam, Ab97224)). The samples were incubated for 30 minutes at25°C with shaking at 300 rpm. The bacteria were then washed with 100 µL HBSS + / + and resuspended in 200 µL HBSS + / +.50µL was transferred to a black 96-well flat-bottom microplate (ThermoScientific, 175325) containing 150µL HBSS + / + to create a 1:4 dilution, and the fluorescence measured using a CLARIOstar plate reader (BMG Labtech). Relative fluorescence values for all samples were recorded and data was analysed using GraphPad CEN-C-P3772PCT Prism. All samples were run in technical triplicates and biological experiments repeated as indicated in the table. The results of this assay are given in Table 2. The results from this assay indicate that Examples 1-6 show recruitment of IgG anti-rhamnose antibodies to the surface of E.coli 700973. Polymyxin B which does not containan antibody recognition element does not show antibody recruitment.Table 2: Anti-Rhamnose IgG recruitment to the surface of E.coli ATCC 700973Compound hIgG1 Recruitment (20µM, average fold changeExperiments over background) Example 1 5.4 n=1Example 2 3.5 n=1Example 3 7.8 n=1Example 4 4.6 n=1Example 5 5.8 n=1Example 6 8.7 n=1Polymyxin B1.0 n=2Control2. Anti-alpha-Gal IgG Recruitment AssayFlow cytometric antibody recruitment assay using anti-Galα1-3Galβ1-4GlcNAc (anti-α-Gal) antibodiesTable 3 shows the recruitment of anti-α-Gal IgG antibodies to the surface of E.coliATCC700973, using a flow cytometry assay. Polymyxin B (PMB) which does not contain an antibody-recognition element, is used as a negative control. To demonstrate that PMB does not recruit antibodies at higher concentrations, data is also shown for PMB using the monoclonal IgM antibody M86. Method: Anti α-Gal IgG recruitment The assays were carried out in polystyrene 96-well U bottom plates (Costar). Frozen stocks of bacteria in mid-exponential phase were thawed, centrifuged and resuspended in LB broth, Miller (Fisher BP1426-500) at OD595 = 0.2. The bacteria was grown to mid-exponential phase (OD595 = 0.4), washed once with Hank's Balanced Salt Solution with calcium and magnesium (HBSS+ / +) and then resuspended in HBSS+ / + at a bacterial D595= 0.6. Bacteria was diluted 1 / 6 and placed in the 96-well U bottom plate. The bacteria were then incubated for 45 minutes with the compound of Example 7 at concentrations of 3.2 and 6.4 µM, CEN-C-P3772PCT Polymyxin B at 2.9 µM or buffer alone (vehicle control), at room temperature and shaking at 450 rpm . The bacteria were then washed three times with 200 µL HBSS+ / +, prior to adding50 µL of a mouse / human chimeric anti-α-Gal IgG3 antibody (clone 22.121, Nozawa, S. etal, (2001), Transplantation 72(1) 147-155) at a final concentration of 80 µg / mL in HBSS+ / +. The samples were incubated for 1 hour at room temperature shaking at 450 rpm. The bacteria were washed three times as above, prior to adding 100 µL of a FITC-labelled anti- human IgG secondary antibody (Goat pAb to Mu IgG-FITC (Invitrogen F2761, lot 2309145))) at 10 µg / mL in HBSS+ / + and incubating at room temperature for 1 hour at 450 rpm. After three final washes with 200 µL HBSS+ / +, the bacteria were resuspended in 200 µL HBSS+ / + and evaluated for anti-α-Gal antibody binding on a Cytoflex flow cytometer (Beckman Coulter).50,000 counts of bacterial particles were sampled and the median fluorescent shift was recorded in the FITC-A channel. Data from all samples were analysed using Kaluza software (Beckman Coulter). All samples were run in technical duplicates. Anti α-Gal IgM recruitment This was carried out as described for the Anti-α-Gal IgG recruitment using mouse / human chimeric anti-alpha Gal IgM antibody M86 (Absolute Antibody Ab00532) at a final concentration of 40 µg / mL in HBSS+ / + as primary antibody and IgM-FITC Goat pAb to Hu IgM FTC (Abcam ab8497) as secondary antibody.Table 3 demonstrates the binding of anti-α-Gal IgG antibodies to the surface of the E. coliATCC 700973, in the presence of Example 7 and Polymyxin B using the flow cytometry assay described above. Data are also shown for Polymyxin B (PMB) as a negative control using anti-α-Gal IgM. The fold shift over background was calculated by dividing the Median FluorescentIntensity (MFI) obtained in the presence of the test compounds by the MFI value obtained in the vehicle controls. i.e., the absence of test compounds. The shift in fluorescence intensity occurs due to the binding event at each end of the molecule.Table 3: Recruitment of anti-α-Gal antibodies to the surface of E. coli ATCC700973Example concentration mIgG3 RecruitmentM86 IgM Recruitment (fold change over (fold change over background) background) 73.2 µM 5.97 6.4 µM 633.3PMB2.9 µM 3.2(control) CEN-C-P3772PCT PMB3.1 µM NT 1.2(control) PMB6.3 µM NT 1.1(control) The results from this assay indicate that Example 7 shows strong recruitment of IgG anti-α-Gal antibodies to the surface of E.coli 700973, in contrast to the negative control PolymyxinB.3. In vitro Renal Cell Toxicity AssayThe renal cell toxicity of the compounds was assessed in an in vitro assay using humanrenal proximal tubule epithelial cells (RPTECs). Human RPTEC cells were seeded into black clear bottom collagen coated 96-well plates in appropriate media. Cells were incubated at 37°C, 5 % CO2 for 72 hr when media was then changed to serum free media. Cells were incubated for a further 24 hr and then compound treated. Test compound was diluted in vehicle and serial dilutions are made to 0.1 % vehicle in RPTEC media. Test compounds at 8 concentrations in triplicate were then incubated for 24 hr. Sertraline and L-buthionine-sulfoximine were simultaneously run as positive controls. At the end of the incubation period, the cells were loaded with the relevant dye, and the plate then scanned using an automated confocal fluorescent cellular imager, ArrayScan® XTI (Thermo Scientific Cellomic s) at 37 °C, 5 % CO2, following which cellular ATP content was measured using CellTiter-Glo® (Promega). A dose-response curve was fitted, and concentration at which 50% maximum effect is observed was determined and reported as the AC50. The results of this assay are shown in Table 4:Table 4: In vitro Renal Cell Toxicity AssayCompound hRPTEC AC50 (µM)Example 1 348Example 2 >500Example 3 >500Example 4 >500Example 5 402Example 6 >500 CEN-C-P3772PCT Compound hRPTEC AC50 (µM)Example 7 >500Reference 13 Example 1 Reference 59 Example 2The results from this assay demonstrate that the compounds of Examples 1 to 7 of theinvention demonstrated significantly reduced cytotoxicity compared with Reference Example1 (i.e. Example 2 from WO 2020 / 201743) and Reference Example 2 (i.e. Example 4 fromWO 2020 / 201743).4. Antimicrobial Susceptibility TestingThe in vitro antimicrobial activity (minimum inhibitory concentration, MIC) of ReferenceExamples 1 and 2 and Examples 1-7 was determined against Escherichia coli ATCC25922,K. pneumoniae NCTC 13438, K. pneumoniae AR039, Pseudomonas aeruginosaATCC27853, Pseudomonas aeruginosa AR064, Acinetobacter baumanii AR296 andAcinetobacter baumanii AR307 by broth microdilution using cation adjusted Mueller-Hintonbroth (caMHB; Becton-Dickinson 212322) according to CLSI guidelines M07-A11. (CLSI M07-A11 (2018). Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically, 11th Edition. Clinical and Laboratory Standards Institute, Wayne, PA 19087, USA).Bacterial inocula were prepared by suspending several well-isolated single colonies in sterilephosphate-buffered saline (PBS) to a density equivalent to a 0.5 McFarland standard. Thesesuspensions were then diluted in caMHB to provide a final bacterial density of 2 to 8 × 105CFU / mL when added to assay plates containing diluted test articles. Assay plates were incubated in air at 37°C for 18 hours, and the MIC determined as the lowest concentration of test articles that inhibited visible bacterial growth. The results of this assay are shown in Table 5:Table 5: Antimicrobial Susceptibility Testing Minimum Inhibitory Concentration(µg / ml) of Examples 1-7 and Reference Examples 1 and 2 CEN-C-P3772PCTExample E.coli K.pneumoniae P.aeruginosa A.baumaniiATCC NCTCAR039 AR064 ATCC AR296 AR30725922 13438 27853Example 1 1 2 1 4 4 1 8Example 2 2 2 4 16 2 4 16Example 3 2 4 4 4 2 4 16Example 4 2 2 4 2 2 4 8Example 5 1 1 1 2 2 2 4Example 6 2 2 2 2 2 4 16Example 7 1 2 2 2 2 2 8Reference1 2 2 2 2 2 4Example 1 Reference1 2 2 4 2 1 4Example 2The results from this assay demonstrate that the compounds of Examples 1 to 7 of theinvention demonstrated equivalent antibacterial activity against the tested strains compared with Reference Example 1 (i.e. Example 2 from WO 2020 / 201743) and Reference Example 2 (i.e. Example 4 from WO 2020 / 201743).5. In vivo efficacy against E. coli thigh infection in miceThe in vivo efficacy of the compound of Examples 5 and 7 was evaluated in a mouse thighinfection model of Escherichia coli ATCC25922. The results are summarized in Table 6.Groups of 5 male C57 / Bl-6 mice were made neutropenic by subcutaneous administration of cyclophosphamide on days -4 (150 mg / kg) and -1 (100 mg / kg). On Day 0, animals wereinoculated intramuscularly with 8 ×104 CFU per thigh of Escherichia coli ATCC25922. At 2h, the CFU count was determined from 5 mice (pre-treatment group). The remaining treatment groups (five per group) were treated with a subcutaneous injection of the test article at +2, +8, +14 and +20 hr post-infection. Twenty-six hours after infection, the mice were euthanized humanely. The thighs of each animal were harvested, homogenized, serially diluted and quantitative tissue burden (CFU / g) measured. Decrease of the total CFU count of each thigh as compared to control counts at 26 hrs post-infection was determined CEN-C-P3772PCT for each dose group. Table 6 shows the geometric mean burden in each of the groups. Treatment with the compound of Example 5 at a total dose of 64 mg / kg (16 mg / kg / dose Q6h) resulted in a 3.23 log10reduction in bacterial counts compared to non-treated control. Treatment with the compound of Example 7 at a total dose of 64 mg / kg (16 mg / kg / dose Q6h), resulted in a 2.99 log10reduction in bacterial counts compared to non-treated control.Table 6: In vivo efficacy versus E. coli ATCC25922 thigh infection in neutropenic miceCompound Total daily doseLog10 change from pre- compound (mg / kg) treatment Example 5 64 3.23Example 7 64 2.99Thus, the compounds of Examples 5 and 7 are both able to reduce the bacterial burden in amouse model of infection.6. In vivo efficacy against K.pneumoniae lung infection in miceThe in vivo efficacy of the compound of Example 7 was evaluated in a mouse lung infectionmodel of Klebsiella pneumoniae. The results are summarized in Table 7. Groups of 5 male C57 / Bl-6 mice were made neutropenic by subcutaneous administration of cyclophosphamide on days -4 (150 mg / kg) and -1 (100 mg / kg). On Day 0, animals were inoculated intranasally with 4 ×104CFU of Klebsiella pneumoniae ATCC43816. At 2 h, the CFU count was determined from 5 mice (pre-treatment group). The remaining treatment groups (five per group) were treated with a subcutaneous injection of the test article at +2, +8, +14 and +20 hr post-infection. Twenty-six hours after infection, the mice were euthanized humanely. The lungs of each animal were harvested, homogenized, serially diluted and quantitative tissue burden (CFU / g) measured. Decrease of the total CFU count of each thigh as compared to control counts at 26 hrs post-infection was determined for each dose group. Table 7 shows the geometric mean burden in each of the groups. Treatment with the compound of Example 7 at a total dose of 72 mg / kg (18 mg / kg / dose Q6h), resultedin 1.65 log10 reduction in bacterial counts compared to non-treated control (vehicle).Treatment with the compound of Example 7 at a total dose of 144 mg / kg (36 mg / kg / dose Q6h), resulted in a reduction in bacterial counts to below pre-treatment levels.Table 7: In vivo efficacy versus K. pneumoniae ATCC43816 infection in neutropenicmice CEN-C-P3772PCTGroup DoseGeometric mean Log10 geometric Log10 Log10 change (mg / kg / burden mean burden change from pre- day) (CFU / g) (CFU / g) from vehicle treatmentPre-treatment n / a 3.40 × 105 5.53 -3.88 0.00Vehicle n / a 2.55 × 109 9.41 0.00 3.88Example 7 72 5.69 × 107 7.75 -1.65 2.22108 1.07 × 107 7.03 -2.38 1.50144 2.57 × 105 5.41 -4.00 -0.12

Claims

1. CEN-C-P3772PCT CLAIMS1. A compound of formula (I) or a pharmaceutically acceptable salt thereof:wherein: F is selected from: rhamnose; or a carbohydrate molecule capable of binding to a human anti-alpha-galactosyl antibody; S2 is selected from: -(CH2)2-O-(CH2)2-NH-CO-CH2-, -(CH2)2-O-(CH2)2-NH-CO-(CH2CH2O)4-(CH2)2-NH-CO-CH2-, –(CH2)3-NH-CO-CH2-, -(CH2CH2O)2-(CH2)2-NH-CO-CH2- or –(CH2)3-CO-NH-(CH2)2-NH-CO-CH2-; Y2 is -O-; R1 represents: C1-6 alkyl, haloC1-6 alkyl or heterocyclyl; Y1 is -CONH-; S1 is -(CH2CH2O)8-(CH2)2-; X1 is -C(=O)-;L represents a cationic anti-microbial peptide selected from a moiety of formula (L):wherein X1represents the point of attachment of L to X1; Z1 represents C1-10 alkyl or benzyl optionally substituted by one or more halogen or haloC1-6 alkyl groups;CEN-C-P3772PCT Z2represents -CH2-CH2-NH2, -CH2-NH2or -CH2-OH; Z3represents benzyl or -CH2-CH(CH3)2;Z4 represents -CH2-CH(CH3)2, -CH(CH3)-CH2CH3, -CH(OH)-CH3 or -CH2CH3; andZ5 represents -CH(OH)-CH3 or -CH2-CH(CH3)2.

2. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inclaim 1, wherein F represents a compound of formula (A):wherein S2refers to the point of attachment to the S2group, or a compound of formula (B):wherein S2 refers to the point of attachment to the S2 group.

3. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inclaim 1 or claim 2, wherein S2 is selected from: -(CH2)2-O-(CH2)2-NH-CO-CH2-, -(CH2)2-O-(CH2)2-NH-CO-(CH2CH2O)4-(CH2)2-NH-CO-CH2- or –(CH2)3-NH-CO-CH2-.

4. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 3, wherein R1 represents C1-6 alkyl, such as C4 alkyl, in particular tertbutyl.

5. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 3, wherein R1 represents haloC1-6 alkyl, such as trifluoromethyl.

6. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 3, wherein R1 represents heterocyclyl, such as a 6 memberedheterocyclyl ring, in particular a 6 membered heterocyclyl ring containing two heteroatoms selected from nitrogen and oxygen, more particularly morpholinyl, especially N-linked morpholinyl.CEN-C-P3772PCT7. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 3, wherein R1 represents tert butyl, trifluoromethyl or morpholinyl,such as N-linked morpholinyl.

8. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 7, wherein Z1represents C1-10alkyl, such as octyl.

9. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 7, wherein Z1represents benzyl optionally substituted by one or more halogen or haloC1-6alkyl groups, such as Z1represents benzyl optionally substituted by one or more halogen (such as fluorine or chlorine) or haloC1-6alkyl (such as trifluoromethyl)groups, in particular Z1 represents 4-flurobenzyl, 4-chlorobenzyl or 4-trifluoromethylbenzyl.

10. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 7, wherein Z1 represents octyl, 4-flurobenzyl, 4-chlorobenzyl or 4-trifluoromethylbenzyl.

11. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 10, wherein Z2 represents -CH2-NH2.

12. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 11, wherein Z3 represents benzyl.

13. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 12, wherein Z4 represents -CH2-CH(CH3)2.

14. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined inany one of claims 1 to 13, wherein Z5 represents -CH(OH)-CH3.

15. A compound of formula (I) or a pharmaceutically acceptable salt thereof as defined in1, which is a compound of formula (I)aor a pharmaceutically acceptable salt thereof:CEN-C-P3772PCT (I)awherein: F is selected from a compound of formula (A):wherein S2refers to the point of attachment to the S2group; or a compound of formula (B):wherein S2 refers to the point of attachment to the S2 group; S2 is selected from: -(CH2)2-O-(CH2)2-NH-CO-CH2-, -(CH2)2-O-(CH2)2-NH-CO-(CH2CH2O)4- (CH2)2-NH-CO-CH2-, or –(CH2)3-NH-CO-CH2-; Y2 is -O-; R1 represents: tert butyl, trifluoromethyl or morpholinyl; Y1 is -CONH-; S1 is -(CH2CH2O)8-(CH2)2-; X1 is -C(=O)-;L represents a cationic anti-microbial peptide selected from a moiety of formula (L):wherein X1 represents the point of attachment of L to X1;CEN-C-P3772PCT Z1represents C1-10alkyl or benzyl optionally substituted by one or more halogen or haloC1-6alkyl groups; Z2represents -CH2-NH2; Z3represents benzyl; Z4represents -CH2-CH(CH3)2; and Z5represents -CH(OH)-CH3.

16. The compound according to claim 1 or a pharmaceutically acceptable salt thereof,which is selected from any one of Examples 1-7.

17. A pharmaceutical composition comprising a compound according to any one ofclaims 1 to 16 or a pharmaceutically acceptable salt thereof.

18. The compound according to any one of claims 1 to 16 or a pharmaceuticallyacceptable salt thereof, or the pharmaceutical composition according to claim 17, for use in therapy.

19. The compound according to any one of claims 1 to 16 or a pharmaceuticallyacceptable salt thereof, or the pharmaceutical composition according to claim 17, for use inthe treatment of a disease or disorder mediated and / or caused by an infective agent.

20. Use of a compound of formula (I) according to any one of claims 1 to 16 or apharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 17, in the manufacture of a medicament for use in the treatment of a disease or disorder mediated and / or caused by an infective agent.

21. A method of treating a disease or disorder mediated and / or caused by an infectiveagent which comprises administering to an individual in need thereof a compound of formula(I) as defined in any one of claims 1 to 16 or a pharmaceutically acceptable salt thereof orthe pharmaceutical composition according to claim 17.

22. A process for preparing a compound of formula (I) according to claim 1 whichcomprises: (a) reacting a compound of formula (II):CEN-C-P3772PCTwherein F, S2, Y2, R1, Y1 and S1 are as defined in claim 1, with a compound of formula (III):where Z1, Z3, Z4 and Z5 are as defined in claim 1, Z2 is either as defined in claim 1 or may represent an appropriately protected derivative of Z2, such as Z2 may contain a protectinggroup, such as Boc, wherein Boc represents an amine protecting group which istertbutoxycarbonyl; followed by(b) deprotecting the Boc protecting groups of the product of step (a) to prepare acompound of formula (I).

23. The process according to claim 22, wherein step (a) comprises reacting the compoundof formula (V) with the compound of formula (VI) in the presence of triethylamine, DMF and HATU.

24. The process according to claim 22 or claim 23, wherein step (b) comprises TFA inDCM and water.

25. A compound of formula (II):CEN-C-P3772PCTwherein F, S2, Y2, R1, Y1 and S1 are as defined in claim 1, or a compound of formula (III):wherein Z3, Z4 and Z5 are as defined in claim 1, Z1 represents benzyl optionally substituted byone or more halogen or haloC1-6 alkyl groups, and Z2 is either as defined in claim 1 or may represent an appropriately protected derivative of Z2, such as Z2 may contain a protecting group, such as Boc, wherein Boc represents an amine protecting group which is tertbutoxycarbonyl.

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