Supramolecular self-associating compounds and uses thereof

Abstract

The present invention concerns a technology platform based on compounds and their capability as supramolecular self-associating amphiphiles (SSA's), nanostructures comprising SSA's, as well as methods of using these, for example in anticancer, antimicrobial, and drug efficacy enhancement applications. The present disclosure further concerns the methods by which the SSA's and nanostructures thereof engage with cell membranes and methods for identifying SSA's.

Description

Case Ref. P319WO IPTector®Supramolecular self-associating compounds and uses thereofTechnical Field

[0001] The present disclosure concerns a technology platform based on compounds and their capability as supramolecular self-associating amphiphiles (SSA's), nanostructures comprising SSA's, as well as methods of using these, for example in anticancer, antimicrobial, and drug efficacy enhancment applications. The present disclosure further concerns the methods by which the SSA's and nanostructures thereof engage with cell membranes and methods for identifying SSA's.Background

[0002] The rise of antimicrobial resistant (AMR) bacterial infections and cancer are two of the greatest health threats facing humanity today. A report generated in 2014 attributed 8.8 million deaths to cancer disease in that year alone, while simultaneously predicting that by the year 2050, ten million people per year will die from the primary effects of AMR. The present impact of AMR was confirmed by a recent study, which reported 4.95 million and 1.27 million deaths from the indirect and direct effects of AMR respectively during 2019. This means that over this time period, AMR was directly responsible for the same number, if not more deaths than those directly attributed to either HIV / AIDs or malaria. This rise in AMR has traditionally been attributed to the misuse of antimicrobial agents, including antibiotics, across multiple sectors. In addition, although we do not yet truly understand the impacts of the COVID-19 pandemic on increased rate of AMR infections, there was a high level of AMR bacterial co-infections confirmed within patients suffering from COVID-19 over the first 18 months of the pandemic.

[0003] Given the gravity of the present and future health threat that AMR poses, there has been a concerted effort to produce novel antimicrobial agents. Due to the lack of financial return, these efforts have been largely shouldered by small and medium sized enterprises. Encouragingly, between 2017-2021, 12 new antibacterial drugs were approved for use in the clinic, with a further 45 in clinical development, 27 of which target WHO priority pathogens. However, despite these success stories, this level of innovation still does not fulfil current or future global need.

[0004] In 2020, there were an estimated 19.3 million new cancer cases identified and almost 10 million cancer-related deaths worldwide. There is no doubt that the burden of cancer is growing globally, due to the aging population, types of risk factors, and rise of resistance to commonly marketed therapeutic agents. In 2020, 313,959 new ovarian cancer cases were reported globally. The number of ovarian cancer-related deaths in this same year was 207,252 (2.1 % of all cancer deaths).Case Ref. P319WO IPTector®The high mortality figures for ovarian cancer are in part due to the fact that approximately 70 % of all diagnoses occur at an advanced stage of the disease, reducing patient survival rates. The high mortality figures are also due to the common use of platinum and taxane-based chemotherapeutic agents, which often lead to complications and resistance, causing 70 % of patients to develop disease recurrence with resistance to these therapeutic agents.

[0005] Due to the expense associated with developing novel drugs, there is much interest in repurposing currently approved therapies to treat different diseases. Interestingly, a number of antimicrobial agents are reported to have off target activity for the treatment of cancer therapy e.g. tetracyclines and fluoroquinolones. However, as well as providing a novel route for the low cost identification of novel cancer agents, the identification of agents with both antimicrobial and anticancer activity may have other advantages for the treatment of cancer patients. Microbial infection is a significant cause of morbidity and mortality in cancer patients, due to general immunosuppression including febrile neutropenia, caused by the effect of the cancer on the human body and / or induced by cancer drug treatment, tumour obstruction or surgery. Of the chemotherapy regimens used to treat ovarian cancer, topotecan, docetaxel and paclitaxel all place a patient at significant risk of febrile neutropenia. Therefore, the art is still in search for a treatment that treats the cancer, treats infectious disease and can overcome resistance in these diseases simultaneously.

[0006] The field of supramolecular chemistry, although young in comparison to the field of medicinal chemistry, is making a considerable effort to exploit our understanding of molecular interactions in the production of next-generation approaches to the design of anticancer and antimicrobial technologies.Summary

[0007] Over this background art the present inventors present a platform of compounds and nanostructures comprising these compounds capable of biological membrane targeting that can enable cell entry to cargo molecules, such as therapeutic agents. These compounds show promising results for antimicrobial, anticancer, and efficacy enhancement applications, in particular for known drugs to which there is limited activity or known resistance. The present disclosure provides means for overcoming major diseases where current pharmaceutical therapies are failing as well as means to re-purpose pharmaceuticals where efficacy / safety is limited by dose and / or delivery route. In some aspects, the compounds of the present disclosure are compatible with target biological membranes, self-associate to produce nanostructures capable of changing structure e.g. by coating the membrane and producing membrane spanning structures in the presence of the target biological membrane and cross the membrane through the formation of pores / channels - or larger membrane disruptionCase Ref. P319WO IPTector®events. The formation of such membrane permeating structures result from the design of the compounds, for example by their ability to engage in complimentary interactions with membrane bound molecules, and the strength of self-associative hydrogen bonding events, the morphology of hydrogen bonding motifs where a plurality of self-associating amphiphiles (SSAs) are supplied simultaneously, the configuration of the compounds' alkyl spacer unit to contain hydrophobic units capable of enhancing hydrophobic self-associative interactions, e.g. pi-pi stacking and / or Van der Waals forces.

[0008] Through balancing one or more or all of these molecular properties the compounds are able to configure into pore or channel-like structures selectively through a biological membrane.

[0009] Accordingly, in one aspect the present disclosure provides a compound of formula (I), X-Z-Y (I),a tautomer, or a pharmaceutically acceptable salt thereof, whereinX, Y, and Z are intramolecularly connected, wherein X is a hydrophobic group, Z is a hydrogen bond donor and acceptor group, and Y is a hydrophilic hydrogen bond acceptor group.

[0010] In a further aspect, a compound capable of self-associating in a liquid medium is provided.

[0011] In a further aspect, a nanostructure is provided comprising a plurality of compounds each independently defined as specified herein.

[0012] In a further aspect, a pharmaceutical composition is provided comprising a compound as defined herein, or a nanostructure as defined herein, and one or more pharmaceutically acceptable excipients.

[0013] In a further aspect, a bilayer membrane, such as a phospholipid membrane, is provided comprising the compound as defined herein, or the nanostructure as defined herein.

[0014] In a further aspect, a cell is provided comprising the membrane as defined herein.

[0015] In a further aspect, a method is provided for treatment of a pathological condition in a subject, the method comprising administering a compound, a nanostructure, or a pharmaceutical composition to the subject.

[0016] In further aspect, a method for inhibiting biofilm on a surface of an object is provided, the method comprising administering a compound or a nanostructure to the surface of the object.

[0017] In further aspect, a process for preparing a nanostructure comprising a plurality of compounds each independently defined as specified herein, the process comprising:a) providing a plurality of compounds each independently defined as specified herein;b) subjecting the compounds to a liquid medium, such as an aqueous medium;c) thereby obtaining the nanostructure.

[0018] In further aspect, a method is provided of selecting a compound capable of or suitable forCase Ref. P319WO IPTector®forming pores and / or channels in a bilayer membrane, for example a phospholipid bilayer membrane, such as a biological cell membrane of a cell, the method comprising one or more or all of:a) Identifying a compound capable of complimentary interacting with phospholipid groups of a biological membrane of a cell;b) Identifying a compound engaging in self-associative hydrogen bonding events;c) selecting a compound as defined herein; and / ord) subjecting a compound to patch clamp analysis to confirm presence of formation of pores and / or channels in the biological cell membrane, andthereby selecting the compound suitable for or capable of forming pores and / or channels in the bilayer membrane.

[0019] In a further aspect, a method is provided of forming pores and / or channels in a bilayer membrane, such as a biological cell membrane of a cell using a nanostructure as defined herein, the method comprising contacting the nanostructure with the bilayer membrane in a liquid medium, for example an aqueous medium, thereby allowing the nanostructure to adhere to and permeate the bilayer membrane forming pores and / or channels.

[0020] In a further aspect, a method is provided of using a compound as defined herein to deliver a therapeutic agent to a biological cell, comprising the steps:a) mixing the compound with the therapeutic agent in a liquid medium;b) forming a nanostructure comprising the compound and the therapeutic agent;c) delivering the nanostructure to a mammal comprising the biological cell, andd) allowing the nanostructure to adhere to and penetrate the biological cell delivering the therapeutic agent to the biological cell.Description of drawings and figures

[0021] The figures included herein are illustrative and simplified for clarity, and they merely show details which are essential to the understanding of the invention, while other details may have been left out. Throughout the specification, claims and drawings the same reference numerals are used for identical or corresponding parts. In the figures and drawing include herein:Figure 1 shows the hypothesised effect of SSAs on target phospholipid membranes. Step 1: Controlled aggregate formation and incorporation of drug cargo. Step 2: SSA aggregate supplied to disease site. Step 3: SSAs target disease cells, increasing the efficacy of drug cargo.Figure 2 shows a graphical representation of the SSA and nanostructures thereof engaging with a cell membrane. Specifically, a) The spontaneously SSA self-assembles under aqueous conditions to form a spherical aggregate, here the structure may also incorporate additionalCase Ref. P319WO IPTector®drug compounds, b) The spherical aggregate arrives at the surface of the target cell membrane, upon adhesion, the SSAs begin to distribute themselves across the outer leaflet of the phospholipid bilayer, c) Upon optimisation of preferential SSA:phospholipid headgroup interactions and increase in local concentration of the SSA, the SSA units now self-associate, producing structures that can permeate the phospholipid bilayer and display 'pore / channel-like' behaviour. However, these structures are not static as the SSA continues to diffuse through the membrane, increasing the local concentration of the SSA on the inner leaflet of the phospholipid membrane and / or diffusing away from this structure into the cell cytoplasm, d) As the local concentration of SSA present increases still further, greater disruption to the phospholipid bilayer is observed, increasing ion flow across this structure, e) As the local concentration of SSA present at the outer or inner leaflet of the phospholipid membrane is reduced, the SSA is no longer able to disrupt the membrane or facilitate the movement of ions, and the phospholipid membrane repairs itself.Figure 3 shows a summary of time spent in different event states upon the addition of SSA30, SSA72, SSA73, SSA56, SSA57, SSA72 + SSA73 (1:1), SSA56 + SSA57 (1:1) at: a) 0.10 mM, b) 0.25 mM, c) 0.38 mM, d) 0.50 mM and e) 1.00 mM. Events were categorised as follows (current, A) 0: A < Event 0 (blue) < 5.00e-ll A, 1: 5.00e-ll A < Event 1 (orange) < 5.00e-10 A, 2: 5.00e-10 A < Event 2 (green) < 4.90e-08 A, 3: 4.90e-08 A < Event 3 (red) <00A. Experiments were conducted over a 1000s ± 5s unless stated. For ease of reference to raw and partially processed data the experiment numbers are given in brackets within this Figure. Planar phospholipid bilayers formed were formed from DPhPC 10 % cholesterol, with Buffer A sealed below, and Buffer B accessible above the bilayer. Buffer A (KCI (489 mM), NaOAc (5 mM), pH 5.5, ionic strength = 500 mM); B (Na2SO4 (167 mM), NaOAc (5 mM), pH 5.5, ionic strength = 500 mM). Stock solutions of SSA(s) were prepared as a DMSO:buffer B 1:50 mixture. Control experiments confirmed the membrane to retain stability in the presence of DMSO at these concentrations. All experiments were conducted with a holding voltage +100 mV.Figure 4 shows a data summary for patch clamp experiments conducted with a holding voltage of +100 mV for: a) SSA73 (0.25 mM); and b) SSA73 (0.50 mM). Events were categorised into the following types: 0: A < Event 0 < 5.00e-ll A, 1: 5.00e-ll A < Event 1 < 5.00e-10 A, 2: 5.00e-10 A < Event 2 < 4.90e-08 A, 3: 4.90e-08 A < Event 3 <00A.Figure 5 shows graphs showing current (A) vs time (s) for: a) SSA73 (0.25 mM), at a holding voltage of +100 mV and b) SSA57 (0.1 mM), at a holding voltage of +100 mV.Figure 6 shows a) Schematic representation of the Cl⁻ / NO₃⁻ antiport assay; b) chloride efflux mediated by SSA72, SSA73 and SSA72 + SSA73 (10 mol% w.r.t. lipid); c) chloride effluxCase Ref. P319WO IPTector®mediated by compounds SSA56, SSA57 and SSA56 + SSA57 (10 mol% w.r.t. lipid); d) an overview of the reciprocal of the EC50values for each compound and racemic mixture to visualise a comparison of the overall efficacies. Where multiple SSAs are supplied in combination, they are supplied as a 1:1 enantiomeric mixture with values reported representing total SSA concentration.Figure 7 shows scatter graphs illustrating the relationship between: a) MAF and PF for SSA6, SSA33, SSA39, SSA74, SSA75 (X) and co-formulations a, d and j (circle); b) MAF (BLACK) or PF (GREY) and zeta potential.Figure 8 shows graphs comparing the PF and MAF (long-dashed lines, error indicated by short-dashed lines) for homogenous solutions of a) SSA6, b) SSA33, c) SSA74 and d) SSA75 with the PF and MAF for the same SSA in a co-formulation (X).Figure 9 shows scatter graphs illustrating the relationship between SSA6, SSA33, SSA39, SSA74, SSA75 and co-formulation a, d and j with MAF (circle) or PF (X) and: a) ECsoat 20 mins for membrane lysis of 1:1 PE: PG vesicles; b) FP values for 1:1 PE: PG vesicles at 1.5 mM. Figure 10 shows the percentage inhibition of P. aeruginosa (PAO1) biofilms by (a) SSA6, SSA33, SSA39, SSA74, SSA75 (n = 3) and, (b) co-formulations a, b, e and j (n > 3) determined through XTT assay. Left to Right: = TBA chloride (control), SSA6, SSA33, SSA39, SSA74, SSA75.Left to right: co-formulation a, b, e, j (0.64 mM only). Error = ± 1 SD.Figure 11 shows percentage inhibition of (a and b) C. albicans (SC5314) biofilms and (c and d) polymicrobial biofilms of C. albicans (SC5314) and P. aeruginosa (PAO1) by (a and c) SSA6, SSA33, SSA39, SSA74, SSA75 (n = 3) and, (b and d) co-formulations a, b and e (n > 3) determined through XTT assay. Left to Right: = TBA chloride (control), SSA6, SSA33, SSA39, SSA74, SSA75. Left to right: co-formulation a, b, e,. Error = ± 1 SD.Figure 12 (a - c) Fluorescent microscopy images of SSA39 taken at different time intervals in the presence of C. albicans, (d - g) Distribution of SSA39 within the (pseudo)hyphae of C. albicans at differing time intervals, (h - k) Distribution of SSA39 along the C. albicans cell body at differing time intervals. Time intervals: d / h = 1 second, e / i = 90 seconds, f / j = 180 seconds, g / k = 335 seconds. Scale bar a - c = 10 pm. Scale bar d - k = 2 pm. Excitation / emission wavelength = 365 / 461 nm.Figure 13 shows the results of the combination synergy titration assay with SSA30 and Cisplatin in A2780 parental cells. Cells were plated in 96-well plates in triplicate and grown under standard conditions for 48 hours followed by treatment with SSA 30 for one hour at sub-cytotoxic concentrations (100 pM, 25 pM, 6.25 pM, 1.56 pM and 0.39 pM) before addition of an 8 point serial dilution of cisplatin. After 96 hours of combination drug treatment, cellsCase Ref. P319WO IPTector®were fixed, stained and analysed using the SRB assay method as described in Example 11. Figure 14 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA30 (0.38 mM), experiment id 219 at +100 mV, for the event between 137.448s and 144.507s.Figure 15 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA30 (0.38 mM), experiment id 220 at +100 mV, for the event between 235.929s and 242.516s.Figure 16 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA30 (0.38 mM), experiment id 220 at +100 mV, for the event between 729.503s and 748.450s.Figure 17 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA30 (1 mM), experiment id 197 at +100 mV, for the event between 399.371s and 407.582s.Figure 18 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA30 (1 mM), experiment id 197 at +100 mV, for the event between 406.802s and 413.374s.Figure 19 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA72 (0.38 mM), experiment id 106 at +100 mV, for the event between 75.441s and 94.311s.Figure 20 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA72 (0.38 mM), experiment id 106 at +100 mV, for the event between 298.541s and 310.056s.Figure 21 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA72 (0.38 mM), experiment id 106 at +100 mV, for the event between 311.812s and 371.659s.Figure 22 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA72 (0.38 mM), experiment id 106 at +100 mV, for the event between 692.435s and 718.469s.Figure 23 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA72 (0.5 mM), experiment id 99 at +100 mV, for the event between 29.681s and 31.531s.Figure 24 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA72 (1 mM), experiment id 85 at +100 mV, for the event between 32.373s and 35.319s.Case Ref. P319WO IPTector®Figure 25 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 21 at +100 mV, for the event between 235.447s and 256.886s.Figure 26 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 21 at +100 mV, for the event between 257.070s and 272.153s.Figure 27 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 21 at +100 mV, for the event between 276.444s and 302.461s.Figure 28 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 24 at +100 mV, for the event between 54.358s and 60.415s.Figure 29 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 24 at +100 mV, for the event between 59.812s and 78.359s.Figure 30 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 24 at +100 mV, for the event between 106.681s and 252.765s.Figure 31 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 24 at +100 mV, for the event between 382.137s and 433.657s.Figure 32 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 24 at +100 mV, for the event between 475.961s and 501.563s.Figure 33 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 24 at +100 mV, for the event between 500.852s and 569.765s.Figure 34 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 30 at +100 mV, for the event between 416.989s and 426.980s.Figure 35 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (0.25 mM), experiment id 59 at +100 mV, for the event between 294.582s and 306.879s.Figure 36 shows patch clamp evidence of phospholipid bilayer channel and / or pore formationCase Ref. P319WO IPTector®displaying current (A) vs time (s) for SSA73 (1 mM), experiment id 69 at +100 mV, for the event between 1455.281s and 1472.842s.Figure 37 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (1 mM), experiment id 80 at +100 mV, for the event between 47.413s and 68.834s.Figure 38 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (1 mM), experiment id 77 at +100 mV, for the event between 40.624s and 53.874s.Figure 39 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA73 (1 mM), experiment id 74 at +50 mV, for the event between 224.693s and 228.745s.Figure 40 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for a racemic mixture of SSA72 and SSA73 (0.1 mM), experiment id 53 at +100 mV, for the event between 312.186s and 376.054s.Figure 41 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA56 (0.25 mM), experiment id 240 at +100 mV, for the event between 40.471s and 70.971s.Figure 42 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA56 (0.38 mM), experiment id 237 at +100 mV, for the event between 38.346s and 47.874s.Figure 43 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA56 (0.5 mM), experiment id 242 at +100 mV, for the event between 30.632s and 47.401s.Figure 44 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for racemic mixture of SSA56 and SSA57 (0.5 mM), experiment id 44 at +100 mV, for the event between 87.792s and 93.593s.Figure 45 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for racemic mixture of SSA56 and SSA57 (0.5 mM), experiment id 46 at +100 mV, for the event between 213.641s and 567.688s.Figure 46 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA57 (0.1 mM), experiment id 42 at +100 mV, for the event between 1023.256s and 1078.537s.Figure 47 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA57 (0.25 mM), experiment id 27 at +100 mV, for theCase Ref. P319WO IPTector®event between 266.025s and 301.898s.Figure 48 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA57 (0.25 mM), experiment id 27 at +100 mV, for the event between 306.870s and 313.125s.Figure 49 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA57 (0.25 mM), experiment id 27 at +100 mV, for the event between 313.267s and 318.659s.Figure 50 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA57 (0.25 mM), experiment id 27 at +100 mV, for the event between 834.543s and 847.024s.Figure 51 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA57 (0.25 mM), experiment id 37 at +100 mV, for the event between 86.506s and 94.427s.Figure 52 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA57 (0.25 mM), experiment id 37 at +100 mV, for the event between 343.620s and 621.194s.Figure 53 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA57 (0.38 mM), experiment id 32 at +100 mV, for the event between 55.308s and 65.671s.Figure 54 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for SSA57 (0.5 mM), experiment id 17 at +100 mV, for the event between 23.793s and 38.938s.Figure 55 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for a racemic mixture of SSA56 and 57 (0.5 mM), experiment id 44 at +100 mV, for the event between 87.792s and 93.593s.Figure 56 shows patch clamp evidence of phospholipid bilayer channel and / or pore formation displaying current (A) vs time (s) for a racemic mixture of SSA56 and 57 (0.5 mM), experiment id 46 at +100 mV, for the event between 213.641s and 567.688s.Incorporation by reference

[0022] All publications, patents, and patent applications referred to herein are incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. In the event of a conflictCase Ref. P319WO IPTector®between a term herein and a term in an incorporated reference, the term herein prevails and controls.Detailed DescriptionThe features and advantages of the present invention is readily apparent to a person skilled in the art by the below detailed description of embodiments and examples of the invention with reference to the figures and drawings included herein.Definitions

[0023] In accordance with this detailed description, the following definitions apply. Note that the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0024] Unless defined otherwise or clearly indicated by context, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0025] The term "amphiphile" as used herein refers to a molecule containing both hydrophilic and hydrophobic regions, enabling it to associate with both aqueous and non-aqueous phases.

[0026] The term "self-associating amphiphile (SSA)" as used herein refers to an amphiphile that can spontaneously assemble into organized structures, such as a nanostructure defined herein, in solution. Self-associating amphiphiles typically exhibit critical aggregation behavior and are influenced by factors such as concentration, temperature, and ionic strength.

[0027] The term "hydrophilic hydrogen bond acceptor group" as used herein refers to a functional group that has an affinity for aqueous environments and can accept hydrogen bonds from donor groups. Examples of hydrophilic hydrogen bond acceptor groups include a carboxylate anion.

[0028] The term "a hydrophobic group" as used herein refers to a nonpolar molecular moiety that repels water and tends to associate with other hydrophobic regions. Hydrophobic groups can include alkyl, aryl, or fluorinated chains, such as methyl (-CH3), ethyl (-C2H5), or perfluorinated alkyl chains, such as trifluoromethyl.

[0029] The term "a hydrogen bond donor and acceptor group" as used herein refers to a functional group that can both donate and accept hydrogen bonds, facilitating intermolecular interactions. Examples of hydrogen bond donor and acceptor groups include hydroxyl (-OH), amide (-CONH2), and carboxyl (-COOH) groups. Non-limiting examples of such functional groups include urea, thiourea, squaramide, guanidinium, and thiouronium, which exhibit strong hydrogen bonding capabilities. These functional groups may be substituted optionally by a lipophilic group, such as an alkyl, for example a C1-C12 alkyl, or a ring system. Further examples encompass structurally related functionalCase Ref. P319WO IPTector®groups capable of similar hydrogen bonding interactions, such as amidinium and other delocalized cationic species with hydrogen bond donor and acceptor properties.

[0030] The term "an electron-withdrawing substituent" as used herein refers to a substituent that withdraws electron density from the atom or moiety to which it is attached through inductive and / or resonance effects, thereby providing net electron-withdrawing character in the relevant molecular context. In certain embodiments, electron-withdrawing character is described functionally by Hammett substituent constants on the standard benzoic-acid scale, for example op>0 and / or om>0, optionally with at least one of op or om being at least +0.05, at least +0.10, or at least +0.20, and in some embodiments at least +0.30 or at least +0.50. Examples of electron-withdrawing substituents include nitro, cyano, halogen, haloalkyl, haloalkoxy, carbonyl-containing groups, sulfur-oxo groups, phosphorus-oxo groups, and electron-poor aryl or heteroaryl groups. Non-limiting examples include nitro (-NO2), cyano (-CN), fluoro, chloro, bromo, iodo, trifluoromethyl (-CF3), pentafluorosulfanyl (-SF5), polyfluoroalkyl (for example -CHF2and -C2F5), polyfluoroalkoxy (for example -OCF3), acyl (for example -C(O)-alkyl and -C(O)-aryl), ester (for example -C(O)O-alkyl), amide (for example -C(O)NH2, -C(O)NH-alkyl, and -C(O)N(alkyl)2), sulfinyl (-S(O)-alkyl), sulfonyl (-SO2-alkyl and -SO2-aryl), sulfonamide (for example -SO2NH2, -SO2NH-alkyl, and -SO2N(alkyl)2), sulfonyl halide (for example -SO2F and -SO2CI), perfluoroalkylsulfonyl (for example -SO2CF3), phosphoryl, phosphonate, phosphinate, pyridyl, pyrimidinyl, pyrazinyl, triazinyl, and pentafluorophenyl. As used in this paragraph, "alkyl" means a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms (C₁-C₆ alkyl), "aryl" means a monocyclic or bicyclic carbocyclic aromatic group having 6 to 14 ring carbon atoms (C₆-C₁₄ aryl), "heteroaryl" means an aromatic ring system having 5 to 14 ring atoms (C₅-C₁₄ heteroaryl) and containing 1 to 4 heteroatoms selected from N, O, and S, "haloalkyl" means alkyl substituted by one or more halogen atoms, "haloalkoxy" means -O-alkyl substituted by one or more halogen atoms, and "optionally substituted" means substituted by one or more substituents selected from halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, nitro, cyano, and oxo, provided the resulting group retains net electron-withdrawing character.

[0031] The term "a lipophilic group" as used herein refers to a substituent that imparts net hydrophobic character to the atom or moiety to which it is attached, thereby increasing relative affinity for non-aqueous or lipid environments and decreasing relative affinity for water. In certain embodiments, lipophilicity is described functionally by a positive hydrophobic substituent constant (Hansch n > 0) and / or by an increase in cLogP and / or logD relative to the corresponding unsubstituted analogue. Examples of lipophilic groups include linear and branched hydrocarbon chains and ring systems. Non-limiting examples include linear or branched alkyl having 1 to 24 carbon atoms (C₁-C24alkyl), alkenyl having 2 to 24 carbon atoms (C2-C24alkenyl), alkynyl having 2 to 24 carbon atoms (C2-Case Ref. P319WO IPTector®C24alkynyl), optionally substituted carbocyclic ring systems having 3 to 12 ring carbon atoms (C3-C12cycloalkyl or cycloalkenyl ring systems), optionally substituted aryl ring systems having 6 to 14 ring carbon atoms (C6-C14aryl ring systems), optionally substituted heterocyclic ring systems having 3 to 12 ring atoms (C3-C12heterocyclyl ring systems), optionally substituted heteroaryl ring systems having 5 to 14 ring atoms (C5-C14heteroaryl ring systems) that retain net lipophilic character, and aralkyl groups having 7 to 20 carbon atoms (C7-C20aralkyl), including fused, bridged, spiro, and bicyclic ring systems, for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl, adamantyl, phenyl, naphthyl, benzyl, and phenethyl. As used in this paragraph, "alkyl" means a straight-chain or branched saturated hydrocarbon group, "alkenyl" means a straight-chain or branched hydrocarbon group containing at least one carbon-carbon double bond, "alkynyl" means a straight-chain or branched hydrocarbon group containing at least one carbon-carbon triple bond, "heterocyclyl" means a nonaromatic monocyclic or bicyclic ring system containing 3 to 12 ring atoms and 1 to 4 heteroatoms selected from N, O, and S, "heteroaryl" means an aromatic ring system containing 5 to 14 ring atoms and 1 to 4 heteroatoms selected from N, O, and S, and "optionally substituted" means substituted by one or more substituents selected from halogen, C₁-C₆ alkyl, C₁-C₆ haloalkyl, C₁-C₆ alkoxy, C₁-C₆ haloalkoxy, and aryl, provided the resulting substituent retains overall lipophilic character.

[0032] The term "nanostructure" as used herein refers to a supramolecular aggregate comprising a plurality (more than one, i.e. two or more) of self-associating amphiphile (SSA) molecules in a liquid medium, in particular an aqueous medium, including without limitation essentially spherical aggregates, vesicular aggregates, elongated aggregates, fibrillar aggregates, and gel fibers.

[0033] The term "anionic hydrophilic hydrogen bond acceptor group" as used herein refers to a hydrophilic hydrogen bond acceptor group carrying a net negative charge under the relevant conditions, including, without limitation, carboxylate, sulfonate, sulfate, and phosphate groups.

[0034] The term "neutral hydrophilic hydrogen bond acceptor group" as used herein refers to a hydrophilic hydrogen bond acceptor group that is overall neutral under the relevant conditions, including, without limitation, carbonyl-containing groups, ethers, sulfoxides, and phosphine oxides.

[0035] The term "dimerization constant (Kdim)" as used herein refers to the equilibrium association constant for dimer formation of an SSA compound, determined by1H NMR dilution experiments, for example in DMSO-d6containing 0.5% H2O at 298 K, with fitting of concentration-dependent chemicalshift data to a self-association model, including an equal-K dimerization model.

[0036] The term "hydrodynamic diameter (dH)" as used herein refers to the hydrodynamic particle size determined by dynamic light scattering (DLS) for an SSA-containing sample under defined conditions, for example in H2O / 5% EtOH at about 5.56 mM and 298 K after annealing, and reported as the principal intensity-distribution peak maximum unless otherwise stated.Case Ref. P319WO IPTector®

[0037] The term "polydispersity index ( PDI )" as used herein refers to the DLS-derived dimensionless measure of sample size distribution breadth, determined under the same conditions used for determination of hydrodynamic diameter (dH), unless otherwise stated.

[0038] The term "zeta potential" as used herein refers to the electrokinetic potential of a nanostructure dispersion determined by electrophoretic light scattering (or equivalent zeta-potential methodology) under defined conditions, for example in H2O / 5% EtOH at a concentration below 6 mM and at 298 K after annealing.

[0039] The term "critical aggregation concentration (CAC)" as used herein refers to the concentration at which aggregation of SSA molecules becomes detectable in a given medium, determined by tensiometry, for example by a pendant-drop method in an aqueous medium, including H2O / 5% EtOH, at about 291 K after annealing.

[0040] The term "surface tension (ST)" as used herein refers to the surface tension value measured by tensiometry under the same conditions used for CAC determination, including the surface tension at CAC unless otherwise specified.

[0041] The term "cLogP" as used herein refers to the calculated logarithm of the octanol / water partition coefficient of a compound, determined using a defined computational tool, for example SwissADME, unless otherwise stated.

[0042] The term "electrostatic surface potential (Emax and Emin)" as used herein refers to, respectively, the maximum positive and minimum negative electrostatic potential values mapped on the molecular surface of an energy-minimized structure, for example obtained after PM6 minimization using Spartan '24 or an equivalent computational protocol.

[0043] The term "pore and / or channel formation (patch-clamp current)" as used herein refers to membrane-conducting activity evidenced by measurable current in a patch-clamp assay, for example using a planar phospholipid bilayer membrane, a nanostructure with compound (SSA) defined herein at a concentration of 1.0 mM or less with respect to the compound, and a holding potential of +100 mV, wherein current (A) is measured as assay output.

[0044] The term "biologically active" as used herein refers to a compound or nanostructure that produces a measurable effect in at least one biological assay described herein, including anti-cancer, anti-microbial, and / or anti-biofilm assays, at the stated assay conditions and concentration range.

[0045] The term "biologically inactive" as used herein refers to a compound or nanostructure that does not produce a measurable effect in the corresponding biological assay under the stated assay conditions and concentration range.

[0046] The term " IC50" as used herein refers to the concentration of a test compound or nanostructure that produces 50% inhibition of the measured response in the specific assay protocolCase Ref. P319WO IPTector®described herein.

[0047] The term " GI50" as used herein refers to the concentration of a test compound or nanostructure that causes 50% growth inhibition relative to untreated control in the specific cellgrowth assay described herein.

[0048] The term " MIC" as used herein refers to the minimum inhibitory concentration that prevents visible microbial growth in the specific microbiological assay described herein.

[0049] The term "bioactive compound" as used herein refers to a compound having measurable biological activity in an assay described herein; in some embodiments, a positively charged bioactive compound is a cationic species comprising at least one carbon atom and at least one cationic center selected from protonated amine, quaternary ammonium, guanidinium, amidinium, imidazolium, and pyridinium.

[0050] The term "tautomer" as used herein refers to constitutional isomers of a compound that interconvert by proton transfer and associated bond rearrangement; all such tautomeric forms are encompassed unless explicitly excluded.

[0051] The term "pharmaceutically acceptable salt" as used herein refers to a salt form of a disclosed compound suitable for pharmaceutical use, formed with a pharmaceutically acceptable counterion; non-limiting cation examples include alkali-metal ions, alkaline-earth-metal ions, ammonium, substituted ammonium, and tetrabutylammonium (TBA), and non-limiting anion examples include chloride, bromide, sulfate, phosphate, acetate, citrate, and mesylate.

[0052] The term "electron-withdrawing substituent" as used herein refers toSSA molecules

[0053] The present disclosure concerns supramolecular self-associating compounds, products deriving therefrom as well as methods to identify these and methods for using them. Supramolecular self-associating amphiphiles (SSAs) are a class of amphiphilic salts and structurally related compounds that are capable of undergoing intermolecular hydrogen bonding events. These events not only influence SSA self-associative properties and any resultant material formation (hydrogels or spherical aggregate), but also the interaction of this class of compounds with other molecular species, such as cell surfaces, in particular phospholipid membranes.

[0054] The present disclosure concerns a series of SSAs designed to, in some embodiments, selectively increase the activity of SSAs against susceptible and resistant biological cells, such as S. aureus and ovarian cancer cells, simultaneously.

[0055] In one embodiment, a compound of formula (I) is provided,X-Z-Y (I),Case Ref. P319WO IPTector®a tautomer, or a pharmaceutically acceptable salt thereof, whereinX, Y, and Z are intramolecularly connected, wherein X is a hydrophobic group, Z is a hydrogen bond donor and acceptor group, and Y is a hydrophilic hydrogen bond acceptor group.

[0056] In one embodiment, the compound of formula (I) is a dimer of the formula Y-Z-X-X-Z-Y, wherein X of one molecule of formula (I) is covalently linked to X of another molecule for formula (I).

[0057] In one embodiment, the compound of formula (I) is an SSA.

[0058] The present disclosure in particular concerns a compound capable of self-associating in a liquid medium.

[0059] In the context of defining a compound capable of self-associating in a liquid medium, i.e. an SSA, the present disclosure sets functional and structural characteristics that are used alone or in combination to define the compound in different embodiments.Functional characteristics

[0060] In some embodiments, the compound disclosed herein is an amphiphile. In some embodiments, the compound is a self-associating amphiphile (SSA).

[0061] Measurement of functional characteristics that can be used to define the compound(s) of the present disclosure is described in detail throughout the examples, and in particular Examples 1 and 2.

[0062] The self-associative properties of the sub-class of SSAs for which Y is a carboxylate were explored in DMSO-dg, as within polar organic solvents, SSAs have previously been shown to produce hydrogen bonded anionic dimers. This enables the quantification of parameters such as SSA anion dimerization constant to be established, which we have previously shown to correlate with critical aggregation concentration (CAC) values obtained from aqueous solutions of that same SSA. Here, an increase in SSA anionic dimerization constant was shown to correlate with a decreasing CAC, when weighted by a calculated LogP value, obtained for the SSA anion hydrophilic moiety.

[0063] Initially, quantitative (Q)1H NMR spectroscopy experiments were undertaken in DMSO-dg 1% DCM, in which the DCM was used as an internal standard to identify the presence of either low or high order SSA self-associated aggregates. Here, low-order SSA self-associated aggregates are defined as those directly observable using traditional solution stateXH NMR spectroscopy (such as the formation of hydrogen bonded anionic dimers), whereas high-order SSA self-associated aggregates are defined as the exact opposite. To determine the presence of low or high-order SSA aggregates, signals corresponding to the anionic or cationic component of the SSA are integrated against those of the internal DCM standard. An apparent 'loss' of signal indicates the presence of larger, NMR silent aggregates, which exhibit solid-like properties and are therefore no longer visible using this experimental technique. As summarised in Example 2, at concentrations < 112 mM, selected SSAsCase Ref. P319WO IPTector®form low order self-associated aggregates in polar organic DMSO-dg solutions. However, 1:1 enantiomeric mixture of selected SSAs also confirm the presence of higher order aggregates, providing evidence to support the hypothesis that the interaction of SSA anion enantiomers influences molecular self-association events in DMSO-dg 1 % DCM. We hypothesise that this is driven by the preferential heterogeneous hydrogen bonded complex formation between the SSA anionic components. Here, the difference in amino acid side chain was also found to influence the relative proportion of the SSA to become incorporated into these higher order SSA aggregates. We hypothesise that the observed increase in SSA to become incorporated into higher order aggregates to be due to a combination favourable pi-pi stacking interactions and decreased steric hinderance, afforded by the change in amino acid R-group.

[0064] In some embodiments, the compound in a polar organic solvent, such as DMSO-dg, exhibits a Kdimof from 15 to 140 M-1, such as from 45 to 135 M-1, such as from 45 to 50 M-1, such as from 50 to 55 M-1, such as from 55 to 60 M-1, such as from 60 to 65 M-1, such as from 65 to 70 M-1, such as from 70 to 75 M-1, such as from 75 to 80 M-1, such as from 80 to 85 M-1, such as from 85 to 90 M-1, such as from 90 to 95 M-1, such as from 95 to 100 M-1, such as from 100 to 105 M-1, such as from 105 to 110 M-1, such as from 110 to 115 M-1, such as from 115 to 120 M-1, such as from 120 to 125 M-1, such as from 125 to 130 M-1, such as from 130 to 135 M-1.

[0065] In some embodiments, the compound in a polar organic solvent, such as DMSO-dg, exhibits a dHof from 0.8 to 2.0 nm, such as from 0.8 to 0.9 nm, such as from 0.9 to 1.0 nm, such as from 1.0 to 1.1 nm, such as from 1.1 to 1.2 nm, such as from 1.2 to 1.3 nm, such as from 1.3 to 1.4 nm, such as from 1.4 to 1.5 nm, such as from 1.5 to 1.6 nm, such as from 1.6 to 1.7 nm, such as from 1.7 to 1.8 nm, such as from 1.8 to 1.9 nm, such as from 1.9 to 2.0 nm. In one particular embodiment the compound in a polar organic solvent, such as DMSO-dg, exhibits a dHof from 1.4 to 1.9 nm.

[0066] In some embodiments, the dimerization constant (Kdim) is determined in a solvent selected from the group consisting of: DMSO-d6, DMSO-d6with 0.5% H2O, and a 1:19 mixture of EtOH: H2O, at a temperature of 298 K.

[0067] Computational calculations can in some embodiments be used to identify primary hydrogen bond donating and accepting sites for the compounds of the present disclosure. These calculations can be conducted in line with studies reported by Hunter using Spartan 20 v1.1.4 (C. A. Hunter, Angew. Chem. Int. Ed., 2004, 43, 5310-5324.). Calculations can in some embodiments be performed using semi-empirical PM6 methods, after energy minimisation calculations, to identify Emaxand Eminvalues. PM6 was used over AMI in line with research conducted by Stewart (J. J. P. Stewart, J. Mol. Model, 2007, 13, 1173-1213).

[0068] In some embodiments, the electrostatic surface potential energy maximum (Emax) andCase Ref. P319WO IPTector®minimum (Emin) are determined by computational calculation as set forth in the Examples. For example, the structure of the compound is first subjected to energy minimization calculations using a semi-empirical method, such as PM6 within a suitable software package. The Emax and Emin values are then obtained from the resulting electrostatic potential maps.

[0069] In some embodiments, the compound exhibits an electrostatic surface potential energy maximum (Emax) of from -20 to -60 kJ·mol-1, such as from -60 to -55 kJ·mol-1, such as from -55 to -50 kJ-mol-1, such as from -50 to -45 kJ-mol-1, such as from -45 to -40 kJ-mol-1, such as from -40 to -35 kJ-mol-1, such as from -35 to -30 kJ-mol-1, such as from -30 to -25 kJ-mol-1, such as from -25 to -20 kJ-mol-1. In some embodiments, the compound exhibits an electrostatic surface potential energy maximum (Emax) of from -46 to -50 kJ·mol-1.

[0070] In some embodiments, the compound exhibits an electrostatic surface potential energy minimum (Emin) of from -650 to -800 kJ·mol-1, such as from -800 to -790 kJ·mol-1, such as from -790 to -780 kJ-mol-1, such as from -780 to -770 kJ-mol-1, such as from -770 to -760 kJ-mol-1, such as from -760 to -750 kJ-mol-1, such as from -750 to -740 kJ-mol-1, such as from -740 to -730 kJ-mol-1, such as from -730 to -720 kJ-mol-1, such as from -720 to -710 kJ-mol-1, such as from -710 to -700 kJ-mol-1, such as from -700 to -690 kJ-mol-1, such as from -690 to -680 kJ-mol-1, such as from -680 to -670 kJ-mol-1, such as from -670 to -660 kJ-mol-1, such as from -660 to -650 kJ-mol-1. In some embodiments, the compound in a polar organic solvent, such as DMSO-d6, exhibits an electrostatic surface potential energy minimum (Emin) of from -700 to -725 kJ·mol-1.

[0071] In some embodiments, the compound exhibits an electrostatic surface potential energy minimum (Emin) as calculated by Spartan '24 of from 0 kJ-mol-1to -250 kJ-mol-1.

[0072] In some embodiments, the compound exhibits an electrostatic surface potential energy maximum (Emax) as calculated by Spartan '24 of from -650 kJ-mol-1to -1050 kJ-mol-1.

[0073] In some embodiments, the compound has a cLogP value as calculated by SwissADME of less than 4.

[0074] In some embodiments, the compound in a polar organic solvent exhibits a dimerization constant (Kdim) of less than 3750 M-1.

[0075] In some embodiments, the compound has a calculated logP obtained using the SwissADME platform or an equivalent art recognized platform. The calculated logP value can be used to theoretically predict and / or quantify the compound's lipophilicity. In some

[0076] In some embodiments, the compound has a calculated logP of from 2.50 to 5.50, such as from 2.50 to 2.75, such as from 2.75 to 3.00, such as from 3.00 to 3.25, such as from 3.25 to 3.50, such as from 3.50 to 3.75, such as from 3.75 to 4.00, such as from 4.00 to 4.25, such as from 4.25 to 4.50, such as from 4.50 to 4.75, such as from 4.75 to 5.00, such as from 5.00 to 5.25, such as from 5.25 to 5.50.Case Ref. P319WO IPTector®In some embodiments, the compound has a logP of from 3.0 to 4.50.

[0077] In some embodiments, logP can be determined experimentally using various methods known in the art, such as the following methods: (1) Shake-Flask Method, involves partitioning a compound between octanol and water, followed by concentration measurement using UV-Vis, HPLC, or GC. (2) pH-Metric Method estimates LogP for ionizable compounds by measuring partitioning at different pH values. (3) Reverse-Phase HPLC correlates a compound's retention time on a hydrophobic column to LogP using reference standards. (4) Electrochemical and solubility-based methods offer alternative approaches.

[0078] In some embodiments, the compound has a logP of from 2.50 to 5.50, such as from 2.50 to 2.75, such as from 2.75 to 3.00, such as from 3.00 to 3.25, such as from 3.25 to 3.50, such as from 3.50 to 3.75, such as from 3.75 to 4.00, such as from 4.00 to 4.25, such as from 4.25 to 4.50, such as from 4.50 to 4.75, such as from 4.75 to 5.00, such as from 5.00 to 5.25, such as from 5.25 to 5.50. In some embodiments, the compound has a logP of from 3.0 to 4.50.

[0079] In some embodiments, the compound has increased capability of self-assembly.Biological activity

[0080] In some embodiments, the compound is biologically active. In some embodiments, the compound is biologically inactive.

[0081] In some embodiments, the biological activity is selected from the group consisting of: anticancer activity, anti-microbial activity, and anti-biofilm activity.

[0082] In some embodiments, the compound has an IC50against a microbial strain of less than 2.0 mM. In some embodiments, the compound has an IC50against a microbial strain of such as from 5 pM to 1.5 mM, such as from 5 pM to 10 pM, such as from 10 pM to 20 pM, such as from 20 pM to 50 pM, such as from 50 pM to 100 pM, such as from 100 pM to 200 pM, such as from 200 pM to 300 pM, such as from 300 pM to 400 pM, such as from 400 pM to 500 pM, such as from 500 pM to 600 pM, such as from 600 pM to 700 pM, such as from 700 pM to 800 pM, such as from 800 pM to 900 pM, such as from 900 pM to 1.0 mM, such as from 1.0 mM to 1.1 mM, such as from 1.1 mM to 1.2 mM, such as from 1.2 mM to 1.3 mM, such as from 1.3 mM to 1.4 mM, such as from 1.4 mM to 1.5 mM.

[0083] In some embodiments, the compound has a MIC against a microbial strain, such as a specific strain disclosed herein, of less than 5.0 mM. In some embodiments, the compound has a MIC against a microbial strain, such as a microbial strain specified herein of from 5 pM to 1.5 mM, such as from 5 pM to 10 pM, such as from 10 pM to 20 pM, such as from 20 pM to 50 pM, such as from 50 pM to 100 pM, such as from 100 pM to 200 pM, such as from 200 pM to 300 pM, such as from 300 pM to 400 pM, such as from 400 pM to 500 pM, such as from 500 pM to 600 pM, such as from 600 pM toCase Ref. P319WO IPTector®700 pM, such as from 700 pM to 800 pM, such as from 800 pM to 900 pM, such as from 900 pM to 1.0 mM, such as from 1.0 mM to 1.1 mM, such as from 1.1 mM to 1.2 mM, such as from 1.2 mM to 1.3 mM, such as from 1.3 mM to 1.4 mM, such as from 1.4 mM to 1.5 mM, such as from 1.5 mM to 1.6 mM, such as from 1.6 mM to 1.7 mM, such as from 1.7 mM to 1.8 mM, such as from 1.8 mM to 1.9 mM, such as from 1.9 mM to 2.0 mM, such as from 2.0 mM to 2.1 mM, such as from 2.1 mM to 2.2 mM, such as from 2.2 mM to 2.3 mM, such as from 2.3 mM to 2.4 mM, such as from 2.4 mM to 2.5 mM, such as from 2.5 mM to 2.6 mM, such as from 2.6 mM to 2.7 mM, such as from 2.7 mM to 2.8 mM, such as from 2.8 mM to 2.9 mM, such as from 2.9 mM to 3.0 mM, such as from 3.0 mM to 3.1 mM, such as from 3.1 mM to 3.2 mM, such as from 3.2 mM to 3.3 mM, such as from 3.3 mM to 3.4 mM, such as from 3.4 mM to 3.5 mM, such as from 3.5 mM to 3.6 mM, such as from 3.6 mM to 3.7 mM, such as from 3.7 mM to 3.8 mM, such as from 3.8 mM to 3.9 mM, such as from 3.9 mM to 4.0 mM, such as from 4.0 mM to 4.1 mM, such as from 4.1 mM to 4.2 mM, such as from 4.2 mM to 4.3 mM, such as from 4.3 mM to 4.4 mM, such as from 4.4 mM to 4.5 mM, such as from 4.5 mM to 4.6 mM, such as from 4.6 mM to 4.7 mM, such as from 4.7 mM to 4.8 mM, such as from 4.8 mM to 4.9 mM, such as from 4.9 mM to 5.0 mM. In some embodiments, the microbial strain is of a prokaryotic microorganism, archaea, or a eukaryotic microorganism.

[0084] In some embodiments, the prokaryotic microorganism, archaea, and / or a eukaryotic microorganism is in planktonic form.

[0085] In some embodiments, the microbial strain is of Pseudomonas aeruginosa, Acinetobacter baumannii, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, or Enterococcus faecium.

[0086] In some embodiments, the gram-positive bacterium is Staphylococcus aureus (S. Aureus), for example methicillin resistant S. aureus (MRSA), Enterococcus faecalis, or Enterococcus faecium.

[0087] In some embodiments, the gram-negative bacterium is Escherichia coli (E. coli), Pseudomonas aeruginosa, or Acinetobacter baumannii.Structural characteristics

[0088] In some embodiments, the compound is of Formula (Al):R2 Formula (Al),wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selectedCase Ref. P319WO IPTector®from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;m is any integer, such as from 1 to 6;n is any integer, such as from 1 to 6;X is selected from the group consisting of: S, O, NH, NH2+, NR10, NHR10+, and SR10+; andwherein each Ri, R2, R3, R4, Rs, Rs, R7, Rs, Re, and R10is independently selected from the group consisting of: hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, such as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino, nitro, cyano, carboxyl, sulfonyl, phosphonyl, or fused with another one of Ri, R2, R3, R4, Rs, Rs, R7, Rs, Re, or Rio of another compound of Formula (Al) forming a dimer between two compounds of Formula (Al); and wherein a pair of RT and Rz, RZand R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of RT and R2, Rz and R3, R3and R4, or R4and R5is bound in formula Al. In some embodiments, the resulting fused bicyclic or tricyclic ring system may be an aromatic or heteroaromatic group as specified specifically herein including the ring systems explicitly disclosed as part of the specific compounds disclosed herein.

[0089] In some embodiments, the substituent Y is a hydrophilic hydrogen bond acceptor group, which can be either an anionic or a neutral hydrophilic hydrogen bond acceptor. When Y is an anionic hydrophilic hydrogen bond acceptor, the counterion A is a cation, which may include a proton (H+), alkali metal ions (e.g., Na+, K+, Li+), alkaline earth metal ions (e.g., Mg2+, Ca2+), ammonium (NH4+), or substituted ammonium groups, such as TBA. Further examples of A include organic cations such as pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, and other heterocyclic or delocalized cationic species, as well as onium salts derived from nitrogen, phosphorus, or sulfur-containing moieties. In cases where Y is a neutral hydrophilic hydrogen bond acceptor, A is absent.

[0090] The values of m and n define the number of repeating units in the structure and are independently selected integers, typically ranging from 1 to 6. In some embodiments, m and n may be combined in any heterogenic combination within this range, allowing for different values of m and nCase Ref. P319WO IPTector®in the same structure. Non-limiting specific examples include (m = 1, n = 1), (m = 1, n = 2), (m = 1, n = 3), (m = 1, n = 4), (m = 1, n = 5), (m = 1, n = 6), (m = 2, n = 1), (m = 2, n = 2), (m = 2, n = 3), (m = 2, n = 4), (m = 2, n = 5), (m = 2, n = 6), (m = 3, n = 1), (m = 3, n = 2), (m = 3, n = 3), (m = 3, n = 4), (m = 3, n = 5), (m = 3, n = 6), (m = 4, n = 1), (m = 4, n = 2), (m = 4, n = 3), (m = 4, n = 4), (m = 4, n = 5), (m = 4, n = 6), (m = 5, n = 1), (m = 5, n = 2), (m = 5, n = 3), (m = 5, n = 4), (m = 5, n = 5), (m = 5, n = 6), (m = 6, n = 1), (m = 6, n = 2), (m = 6, n = 3), (m = 6, n = 4), (m = 6, n = 5), (m = 6, n = 6). In some embodiments, m and n may be restricted to a narrower range, such as 1 to 4, depending on the desired physicochemical properties of the compound.

[0091] In some embodiments, the atom X is includes sulfur (S), oxygen (O), and nitrogen-containing moieties such as NH, NH2+, NR10, NHR10+, and sulfur-containing moieties such as SR10+. In some embodiments, X may preferentially be oxygen (O) or sulfur (S) to enhance specific electronic or steric properties of the compound.

[0092] In some embodiments, the substituents R1, R2, R3, R4, R5, Rs, R7, Rs, Rs, and R10include:Unsubstituted or substituted aliphatic groups, including linear or branched alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, neopentyl, hexyl, heptyl, octyl, nonyl, decyl) and cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl), optionally substituted with one or more functional groups such as hydroxyl, halogen, amino, carboxyl, or cyano.Unsaturated hydrocarbon groups, including alkenyl (e.g., vinyl, allyl, butenyl, pentenyl, hexenyl) and alkynyl (e.g., ethynyl, propargyl, butynyl, pentynyl), optionally substituted with halogen, hydroxyl, cyano, nitro, or sulfonyl groups.Aromatic groups, including aryl (e.g., phenyl, naphthyl, anthracenyl, biphenyl) and heteroaryl (e.g., pyridyl, thiophenyl, furanyl, benzofuranyl, benzothiophenyl, quinolinyl, isoquinolinyl, indolyl, benzothiazolyl, benzimidazolyl, thiazolyl, oxazolyl, pyrazolyl, pyrimidinyl, pyridazinyl, triazolyl, tetrazolyl), optionally substituted with -e alkyl groups (e.g., methyl, ethyl, propyl), halo groups, hydroxyl, cyano, nitro, sulfone, or amide functionalities.Heterocyclic groups, including saturated, partially saturated, or fully conjugated heterocycles containing one or more nitrogen, oxygen, or sulfur atoms (e.g., pyrrolidine, piperidine, morpholine, thiomorpholine, azetidine, pyrrolizidine, oxetane, tetrahydrofuran (THF), tetrahydropyran, dioxane, thiazolidine, imidazolidine, piperazine, triazocane), optionally substituted with alkyl, aryl, halo, hydroxyl, amino, carboxyl, cyano, or sulfone groups.Functionalized groups, including acyl (e.g., formyl, acetyl, propionyl, butyryl, isobutyryl, benzoyl, trifluoroacetyl, pivaloyl), alkoxy (e.g., methoxy, ethoxy, propoxy, butoxy, isopropoxy, tert-butoxy), aryloxy (e.g., phenoxy, naphthoxy, biphenyloxy, benzyloxy), amino (e.g., primary amines (-NH2),Case Ref. P319WO IPTector®secondary amines (-NHR), tertiary amines (-NR2)), imino (-C=NH, -C=NR), thioalkyl (e.g., methylthio, ethylthio, propylthio, butylthio), thioaryl (e.g., phenylthio, benzylthio, thiophenyl), silyl (e.g., trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldimethylsilyl (TBDMS), triisopropylsilyl (TIPS), bis(trimethylsilyl)methyl), and phosphino (-PR2, e.g., diphenylphosphino, triethylphosphino, triphenylphosphino) groups.Halogenated groups, including haloalkyl groups (e.g., trifluoromethyl (-CF3), pentafluoroethyl (-C2F5), heptafluoropropyl (-C3F7), nonafluorobutyl (-C4F9), chloromethyl (-CH2Cl), bromomethyl (-CH2Br), iodomethyl (-CH2I), difluoromethyl (-CHF2), fluoromethoxy (-OCF3), perfluoroisopropyl (-C(CF3)2H)), perfluoroalkyl (-CnF2n+l, e.g., perfluorobutyl, perfluorohexyl, perfluorooctyl), and fluorinated ether (-OCF3, -OC2F5, -OC3F7, -OC4F9) groups. Additional halogenated substituents include SF5(pentafluorosulfanyl), SF5O (pentafluorosulfanyl oxide), SF5CF3(pentafluorosulfanyl trifluoromethyl), SF5OCF3(pentafluorosulfanyl oxytrifluoromethyl), SF5-aryl, SF5-heteroaryl, and SF5-aliphatic groups, and one or more halogens (e.g., F, Cl, Br, I).Fused or linked groups, where any two or more R-groups may be covalently fused, forming a dimer of compounds having Formula (Al). Further, in any of the Formulas disclosed herein, a pair of RT and Rz, Rzand R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of RT and Rz, Rzand R3, R3and R4, or R4and R5is bound in the specific Formula, such as Formula Al, A2, Bl, or B2.

[0093] In some embodiments, the compound is of Formula (A2):‘3 R5■2 NRi Formula (A2);wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selected from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;n is any integer, such as from 1 to 6;X is selected from the group consisting of: S, O, NH, NHz+, NRio, NHR10+, and SR10+; andwherein each Ri, Rz, R3, R4, Rs, Rs, R7, and R10is independently selected from the group consisting of:Case Ref. P319WO IPTector®hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, such as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino, nitro, cyano, carboxyl, sulfonyl, phosphonyl, or fused with another one of Ri, R2, R3, R4, Rs, Rs, R7, or RM of another compound of Formula (Al) forming a dimer between two compounds of Formula (A2); and wherein a pair of RT and Rz, RZ and R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of RT and Rz, Rzand R3, R3and R4, or R4and R5is bound in formula A2.

[0094] In some embodiments, a compound of Formula (A3) is provided:Formula (A3);wherein each occurrence of Ri is independently selected from the group consisting of a lipophilic substituent and an electron-withdrawing substituent;wherein n is 1, 2, or 3,for example wherein the lipophilic substituent is selected from the group consisting of C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C12 cycloalkyl, aryl, a non-aromatic ring system, a haloalkyl, such as C1-C24 haloalkyl, such as CF3, and SF5;for example wherein the electron-withdrawing substituent is selected from the group consisting of CF3, SF5, cyano, and halogen;wherein X is selected from the group consisting of O, S, guanidino, optionally substituted guanidinium, and optionally substituted thiouronium;wherein R3 is selected from the group consisting of hydrogen, C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, an aromatic ring system, and a non-aromatic ring system;wherein Y is selected from the group consisting of a phosphate, sulfate, and carboxylate; and wherein A is selected from the group consisting of a proton, and a carbon-containing cation.

[0095] In some embodiments, the compound is of Formula (A3):Case Ref. P319WO IPTector®wherein Rzis selected from the group consisting of a lipophilic substituent and an electronwithdrawing substituent;wherein n is 1, 2, or 3,for example wherein the lipophilic substituent is selected from the group consisting of C1-C24alkyl, C2-C24alkenyl, C2-C24 alkynyl, C3-C12 cycloalkyl, aryl, a non-aromatic ring system, a haloalkyl, such as C1-C24 haloalkyl, such as CF3, and SF5;for example wherein the electron-withdrawing substituent is selected from the group consisting of CF3, SF5, cyano, and halogen;wherein X is selected from the group consisting of O, S, guanidino, optionally substituted guanidinium, and optionally substituted thiouronium;wherein R3 is selected from the group consisting of hydrogen, C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, an aromatic ring system, and a non-aromatic ring system;wherein Y is selected from the group consisting of a phosphate, sulfate, and carboxylate; and wherein A is selected from the group consisting of a proton, and a carbon-containing cation.

[0096] In some embodiments, R2is selected from the group consisting of: C1-C24alkyl, C2-C24alkenyl, C2-C24 alkynyl, C3-C14cycloalkyl, aryl, heteroaryl, C3-C14heterocyclyl, C1-C10haloalkyl, halogen, cyano, CF3, and SF5; wherein each of said C1-C24alkyl, C2-C24alkenyl, C2-C24alkynyl, C3-C14cycloalkyl, aryl, heteroaryl, and C3-C14heterocyclyl groups is optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, hydroxy, C1-C6alkoxy, C1-C6haloalkoxy, amino, mono(C1-C6alkyl)amino, di(C1-C6alkyl)amino, cyano, CF3, SF5, -C(=O)ORa, -C(=O)NRaRb, -SO2Ra, and -SO2NRaRb; wherein any C1-C24alkyl, C2-C24alkenyl, or C2-C24alkynyl group in R2is optionally interrupted by 1 or 2 linkages independently selected from -O-, -S-, -NRa-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NRa-, and -NRaC(=O)-; wherein Ra and Rb are independently selected from H and C1-C6alkyl.

[0097] In a further embodiment, R2is selected from the group consisting of: C1-C12alkyl, C2-C12alkenyl, C2-C12alkynyl, C3-C10cycloalkyl, aryl, heteroaryl, -Ce haloalkyl, CF3, SF5, and cyano; wherein each of said C1-C12alkyl, C2-C12alkenyl, C2-C12alkynyl, C3-C10cycloalkyl, aryl, and heteroaryl groups is optionally substituted with 1 or 2 substituents independently selected from halogen, cyano, CF3, SF5, C1-C4alkoxy, -C(=O)ORa, and -C(=O)NRaRb; wherein any C1-C12alkyl, C2-C12alkenyl, or C2-C12alkynyl group in R2is optionally interrupted by 1 linkage selected from -O-, -C(=O)O-, and -C(=O)NRa-; wherein C1-C12alkyl includes linear and branched alkyl.

[0098] In some embodiments, the compound is of Formula (A3):Formula (A3);Case Ref. P319WO IPTector®wherein each occurrence of Ri is independently selected from the group consisting of a lipophilic substituent and an electron-withdrawing substituent;wherein n is 1, 2, or 3,for example wherein the lipophilic substituent is selected from the group consisting of C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C12 cycloalkyl, aryl, a non-aromatic ring system, a haloalkyl, such as C1-C24 haloalkyl, such as CF3, and SF5;for example wherein the electron-withdrawing substituent is selected from the group consisting of CF3, SF5, cyano, and halogen;wherein X is selected from the group consisting of O, S, guanidino, optionally substituted guanidinium, and optionally substituted thiouronium;wherein R3 is selected from the group consisting of hydrogen, C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, an aromatic ring system, and a non-aromatic ring system;wherein Y is selected from the group consisting of a phosphate, sulfate, and carboxylate; and wherein A is selected from the group consisting of a proton, and a carbon-containing cation.

[0099] In some embodiments, A is selected from the group consisting of: a proton (H+), a substituted ammonium group, preferably tert-butyl ammonium (TBA), and a bioactive compound, for example a proton or TBA.

[0100] In some embodiments, when Y carries a negative charge, A is present in an amount that provides overall electroneutrality.

[0101] In some embodiments, n is 1.In some embodiments, each occurrence of Rl is independently selected from the group consisting of: C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C14 cycloalkyl, aryl, heteroaryl, C3-C14 heterocyclyl, C1-C10 haloalkyl, halogen, cyano, CF3, and SF5; wherein each of said C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C14 cycloalkyl, aryl, heteroaryl, and C3-C14 heterocyclyl groups is optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, hydroxy, C1-C6 alkoxy, C1-C6 haloalkoxy, amino, mono(Cl-C6 alkyl)amino, di(Cl-C6 alkyl)amino, cyano, CF3, SF5, -C(=O)ORa, -C(=O)NRaRb, -SO2Ra, and -SO2NRaRb; wherein any C1-C24 alkyl, C2-C24 alkenyl, or C2-C24 alkynyl group in Rl is optionally interrupted by 1 or 2 linkages independently selected from -O-, -S-, -NRa-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NRa-, and -NRaC(=O)-; wherein Ra and Rb are independently selected from H and C1-C6 alkyl.

[0102] In a further embodiment, each occurrence of Rl is independently selected from the group consisting of: C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C3-C10 cycloalkyl, aryl, heteroaryl, C1-C6 haloalkyl, CF3, SF5, and cyano; wherein each of said C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C3-Case Ref. P319WO IPTector®CIO cycloalkyl, aryl, and heteroaryl groups is optionally substituted with 1 or 2 substituents independently selected from halogen, cyano, CF3, SF5, C1C1-C4alkoxy, -C(=O)ORa, and -C(=O)NRaRb; wherein any C1-C12 alkyl, C2-C12 alkenyl, or C2-C12 alkynyl group in R1 is optionally interrupted by 1 linkage selected from -O-, -C(=O)O-, and -C(=O)NRa-.

[0103] In some embodiments, R3 is selected from the group consisting of: H, -C24 alkyl, C2-C24alkenyl, C2-C24alkynyl, C3-C14cycloalkyl, C6-C14aryl, C7-C20aryl(C1-C6)alkyl, 5- to 14-membered heteroaryl, and 3- to 14-membered non-aromatic heterocyclyl; wherein C1-C24alkyl includes linear and branched alkyl; wherein each R3other than H is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen, hydroxy, oxo, cyano, nitro, amino, mono(C1-C6alkyl)amino, dif -Cg alkyl)amino, -Cg alkyl, C C6haloalkyl, -Cg alkoxy, -Cg haloalkoxy, C C6alkylthio, CF3, SF5, carboxyl, C C6alkoxycarbonyl, C C6alkylcarbonyl, -C(=O)NH2, -C(=O)NH(C1-C6alkyl), -C(=O)N(C1-C6alkyl)2, -SO2(C1-C6alkyl), -SO2NH2, -SO2NH(C1-C6alkyl), -SO2N(C1-C6alkyl)2, -P(=O)(OH)2, and -P(=O)(O(C1-C6alkyl))2; wherein any C1-C24alkyl, C2-C24alkenyl, or C2-C24alkynyl portion of R3is optionally interrupted by 1 or 2 linkages independently selected from -O-, -S-, -NH-, -N(C1-C6alkyl)-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH-, -NHC(=O)-, -C(=O)N(C1-C6alkyl)-, and -N(C C6alkyl)C(=O)-.

[0104] In a further embodiment, R3is selected from the group consisting of H, C1-C12alkyl, C2-C12alkenyl, C2-C12alkynyl, C3-C10cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, C7-C16aryl(C1-C4)alkyl, 5- to 10-membered heteroaryl, and 5- to 10-membered non-aromatic heterocyclyl; wherein C1-C12alkyl includes linear and branched alkyl, including isobutyl; wherein C7-C16aryl(C1-C4)alkyl includes benzyl, 1-naphthylmethyl, and 2-naphthylmethyl; wherein each R3group other than H is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen, cyano, CF3, SF5, C1-C4alkyl, C1-C4haloalkyl, C1-C4alkoxy, C1-C4haloalkoxy, amino, mono(C1-C4alkyl)amino, di(C1-C4alkyl)amino, -C(=O)O(C1-C4alkyl), -C(=O)NH2, -C(=O)NH(C1-C4alkyl), -SO2(C1-C4alkyl), and -SO2NH2; and wherein any alkyl, alkenyl, or alkynyl portion of R3is optionally interrupted by 1 linkage selected from -O-, -C(=O)O-, and -C(=O)NH-.

[0105] In a further embodiment, R3is selected from the group consisting of isobutyl, sec-butyl, tertbutyl, benzyl, 1-naphthylmethyl, and 2-naphthylmethyl, each optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen, C1-C4alkyl, C1-C4alkoxy, CF3, SF5, and cyano.

[0106] In some embodiments, each occurrence of R1 is independently selected from the group consisting of C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C3-C12 cycloalkyl, aryl, heteroaryl, non-aromatic ring system, CF3, and SF5, cyano, nitro, halogen, C(=O)C1-C6 alkyl, C(=O)O-C1-C6 alkyl, C(=O)NH2, C(=O)NH(C1-C6 alkyl), SO2-C1-C6 alkyl, SO2NH2, and SO2NH(C1-C6 alkyl).- 27 -Case Ref. P319WO IPTector®

[0107] In some embodiments, X is selected from the group consisting of O, S, guanidino, and thiouronium.

[0108] In some embodiments, X is optionally substituted guanidinium, or optionally substituted thiouronium. In these embodiments, the optional substituent may be a proton but may also be any lipophilic group or ring system. In some embodiments, the substituent is a C1-C12 alkyl either brancher or unbranched.

[0109] In some embodiments, a compound of Formula (A4) is provided;wherein R3 is isobutyl or benzyl.

[0110] In some embodiments, the compound is of Formula (Bl):AFormula (Bl);wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selected from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;m is any integer, such as from 1 to 6;n is any integer, such as from 1 to 6; andwherein each Ri, R2, R3, R4, Rs, Rs, R7, Rs, Re, and R10is independently selected from the group consisting of: hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, such as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino, nitro, cyano, carboxyl, sulfonyl,Case Ref. P319WO IPTector®phosphonyl, or fused with another one of Ri, R2, R3, R4, Rs, Rs, R7, Rs, Re, or Rio of another compound of Formula (Bl) forming a dimer between two compounds of Formula (Bl); and wherein a pair of RT and R2, Rz and R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of RT and R2, R2and R3, R3and R4, or R4and R5is bound in formula Bl.

[0111] In some embodiments, the compound is of Formula (B2):R3‘2 N N NA 7nYH Formula (B2);wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selected from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;m is any integer, such as from 1 to 6;n is any integer, such as from 1 to 6; andwherein each Ri, R2, R3, R4, Rs, Rs, R7, Rs, Re, and R10is independently selected from the group consisting of: hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, such as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino, nitro, cyano, carboxyl, sulfonyl, phosphonyl, or fused with another one of Ri, R2, R3, R4, Rs, Rs, R7, Rs, Re, or Rio of another compound of Formula (B2) forming a dimer between two compounds of Formula (B2); and wherein a pair of RT and Rz, R2and R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of RT and R2, Rz and R3, R3and R4, or R4and R5is bound in formula B2.

[0112] In some embodiments, Y comprises a carboxylate anion.

[0113] In some embodiments, Y comprises an anion, such as an anion selected from a carboxylateCase Ref. P319WO IPTector®anion, a sulfonate anion, and a phosphate anion. In some embodiments, this definition of Y is selected in combination with one or more functional characteristics of the compound as specified herein.

[0114] The design of these SSAs: (i) builds on structure activity relationship analysis, which suggests that the presence of a carboxylate functionality within the structure of the SSA anion increases the strength of preferential intermolecular interactions (ii) explores the effects of chirality on SSA (host) biological activity, as phospholipid headgroups can act as chiral guest species, during initial biological membrane interaction events; (iii) use the presence of a hydrophobic amino acid R-group to increase the stability of any SSA transmembrane structures within the hydrophobic portions of the phospholipid bilayer through shielding of the SSA anion hydrophilic moieties, and / or introducing n-n stacking interactions.

[0115] In some embodiments, A comprises tetrabutylammonium (TBA).

[0116] In some embodiments, Z is a squaramide.

[0117] In some embodiments, R6 and / or R7 are a hydrophobic group.

[0118] In some embodiments, one of R6 and R7 is a hydrophobic group and one is hydrogen.

[0119] In some embodiments, one of R6 and R7 is a branched or straight alkyl chain, such as isobutyl, or an aralkyl, such as benzyl.

[0120] In some embodiments, at least 2, such as at least 3, such as at least 4 of Rl, R2, R3, R4, and R5 is hydrogen.

[0121] In some embodiments, one of Rl, R2, R3, R4, and R5 is a trifluoroalkyl, such as trifluoromethyl.

[0122] In some embodiments, the compound is chiral.

[0123] In some embodiments, the compound is essentially a single enantiomer or a mixture of enantiomers, for example essentially a 1:1 mixture of enantiomers.

[0124] In some embodiments, the compound is of formula A2, Al, Bl, or B2, and wherein one or more of R6, R7, R8, and R9 are independently selected from H and a hydrophobic group, wherein at least one of R6, R7, R8, and R9 is a hydrophobic group.

[0125] In some embodiments, the compound is not any one of:Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®

[0126] In some embodiments, the compound is selected from the group consisting of:Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®TBATBACase Ref. P319WO IPTector®55TBACase Ref. P319WO IPTector®86Case Ref. P319WO IPTector®L120 / D121 L118 / D119L126 / D127 L124 / D125 L130 / D131139,o-TBACase Ref. P319WO IPTector®

[0128] In some embodiments, the compound is selected from the group consisting of:, and a stereoisomeric mixture thereof.

[0129] In some embodiments, the compound is selected from the group consisting of:Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®

[0131] In some embodiments, the the compound is selected from the group consisting of:TBAH Hisomer L 56 / isomer D 57 Isomer L 72 / lsomer D 73Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®and a salt thereof, such as a pharmaceutically acceptable salt thereof.

[0132] In some embodiments, the plurality of compounds is selected from the group consisting of:Case Ref. P319WO IPTector®o' °’TBA(SSA6, 33, 39, 74, 75, 56, 57, 72, 73, 48), and stereoisomeric mixtures thereof.Nanostructure of SSAs

[0133] In some embodiments, a nanostructure is provided comprising a plurality (more than one) of compounds each independently defined as specified herein.

[0134] In some embodiments, the plurality of compounds self-associate in a liquid medium, such as an aqueous medium, thereby forming the nanostructure.Function of nanostructures

[0135] In the following, embodiments are provides describing functional aspects of the nanostructures of the present disclosures. It is an aspect of the disclosure, that any function described herein also provides a disclosure of a method of carrying out said function.

[0136] In some embodiments, the nanostructure is configured for or capable of interacting with a biological membrane and adhering to and / or penetrating the biological membrane to form pores and / or channels in the biological membrane.

[0137] In some embodiments, the adhering to and / or penetrating the biological membrane increases the susceptibility of the biological membrane to a therapeutic agent and / or increases the permeability of the therapeutic agent across the biological membrane when contacted with the therapeutic agent.

[0138] In some embodiments, the nanostructure is capable of or configured for acting as a supramolecular host to one or more phospholipid headgroups of a biological membrane acting as supramolecular guests.

[0139] In some embodiments, the nanostructure is capable of forming or forms a supramolecular host:guest complex with one or more phospholipid headgroups of a biological membrane when contacted with one or more phospholipid headgroups of a biological membrane.

[0140] In some embodiments, the nanostructure is capable of or configured for absorbing or adsorbing a therapeutic agent, such as a small molecule.

[0141] In some embodiments, the nanostructure is capable of or configured for absorbing orCase Ref. P319WO IPTector®adsorbing a therapeutic agent, such as a small molecule, and further capable of or configured for delivering the therapeutic agent to a biological cell.

[0142] In some embodiments, the nanostructure is capable of or configured for forming pores and / or channels in a bilayer membrane, for example a phospholipid membrane, for example a biological cell membrane, preferably as evidenced by patch clamp analysis.

[0143] In some embodiments, the nanostructure is capable of or configured for forming pores and / or channels in a bilayer membrane to enable ion-transport across the bilayer membrane, for example by creating an ion-channel. In some embodiments, the ion-channel is dynamic and may disappear after a predefined amount of time.

[0144] In some embodiments, the nanostructure is capable of or configured for forming pores and / or channels in a bilayer membrane to enable ion-transport across the bilayer membrane.

[0145] In some embodiments, the nanostructure is capable of or configured for forming pores and / or channels in a bilayer membrane of a vesicle to to enable ion, such as anion, efflux across the bilayer membrane.

[0146] In some embodiments, the nanostructure has a membrane permeation (PF) of above 0, such as approximately 1, such as 1, or above 1, such as from 1 to 2 or to above 2.

[0147] In some embodiments, the nanostructure has a membrane adhesion factor (MAF) of 0.5 or less.

[0148] In some embodiments, the nanostructure has a membrane adhesion factor (MAF) of less than 0.5.

[0149] In some embodiments, the nanostructure has a MAF of 0.5 or less, and a zeta potential of between -70 mV and -80 mV.

[0150] In some embodiments, the nanostructure is provided characterized by a MAF of 0.5 or less and a PAF of 1 or higher, such as a PAF of 1 to 2 or of 1 to 3.

[0151] In some embodiments, the membrane adhesion factor (MAF) and membrane permeation factor (PF) are determined by1H CPMG NMR spectroscopy as set forth in the Examples. For example, the compound or nanostructure is added to a solution of pre-formed phospholipid vesicles, such as 1:1 PE: PG vesicles, in the presence of a paramagnetic relaxation enhancement agent, such as gadodiamide.1H CPMG NMR spectra are collected, and the MAF and PF are calculated based on changes in the peak intensities of the compound's aromatic CH resonances.

[0152] In some embodiments, the capability of a compound or nanostructure to form pores and / or channels is determined by patch-clamp analysis as set forth in the Examples. For example, a planar phospholipid bilayer, such as one formed from DPhPC with 10% cholesterol, is established on a patchclamp chip, separating two buffer solutions. A holding voltage, such as +100 mV, is applied across theCase Ref. P319WO IPTector®bilayer. The compound or nanostructure is added to the solution on one side of the bilayer, and the resulting change in electrical current (A) across the membrane is measured over time to detect the formation of ion transport events.Structure of nanostructures

[0153] In some embodiments, the plurality of compounds comprises the same or different compounds.

[0154] In some embodiments, the plurality of compounds is independently selected from any compound specifically disclosed in

[0127] ,

[0155] In some embodiments, a nanostructure is provided comprising any combination of compounds disclosed herein, such as any combination of at least two compounds specifically disclosed in

[0127] , such as specifically two compounds disclosed therein.

[0156] In some embodiments, the plurality of compounds is selected from the group consisting of:Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®

[0157] In some embodiments, the plurality of compounds is selected from the group consisting of:Case Ref. P319WO IPTector®mixtures thereof.

[0158] In some embodiments, the plurality of compounds is selected from the group consisting of:and stereoisomeric mixtures thereof.

[0159] In some embodiments, the nanostructure provided herein comprises two different compounds. In some embodiments, the nanostructure provided herein comprises at least two different compounds.

[0160] In some embodiments, the nanostructure provided herein comprises a combination (two) of compounds selected from the group consisting of:Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®

[0161] In some embodiments, the nanostructure comprises a mixture of enantiomers, for example in a ratio of from 30 to 70, such as from 35 to 65, such as from 40 to 60, such as for example approximately 50:50.

[0162] In some embodiments, the plurality of compounds comprises essentially a single enantiomer.

[0163] In some embodiments, the nanostructure is capable of aggregating into an essentially spherical form, or a gel fiber, or a combination thereof.

[0164] In some embodiments, the nanostructure is essentially spherical when in a liquid medium, such as an aqueous medium.Parameters

[0165] In the following paragraphs, the nanostructure of the present disclosure is defined by a set of parameters. These parameters can be obtained as set out in the present examples, including in some embodiments the use of solvent(s) and / or concentration(s) essentially as specified in the Examples. In some embodiments, the nanostructure is defined by a combination of more than one of the following parameters.

[0166] In some embodiments, the parameters disclosed herein are produced as set out in the Examples, in particular Example 2. In some embodiments, the parameters, such as physicochemical parameters are obtained for the nanostructure(s) in a H2O 5% EtOH or D2O 5 % EtOH solution. In some embodiments, CAC is derived at approximately 291 K from surface tension measurements.

[0167] In some embodiments, the Critical Aggregation Concentration (CAC) and Surface Tension (ST)Case Ref. P319WO IPTector®are determined by tensiometry as set forth in the Examples. For example, samples are prepared in an aqueous medium, such as an H2O / 5% EtOH solution, through serial dilution and are subjected to an annealing process. The surface tension of each sample is then measured at approximately 291 K using a suitable method, such as the pendant drop method, and the CAC is determined as the concentration at which a distinct change in the slope of surface tension versus concentration is observed.

[0168] In some embodiments, the nanostructure has a hydrodynamic diameter, dHof from 90 to 1000 nm, such as from 90 nm to 100 nm, such as from 100 nm to 110 nm, such as from 110 nm to 120 nm, such as from 120 nm to 130 nm, such as from 130 nm to 140 nm, such as from 140 nm to 150 nm, such as from 150 nm to 160 nm, such as from 160 nm to 170 nm, such as from 170 nm to 180 nm, such as from 180 nm to 190 nm, such as from 190 nm to 200 nm, such as from 200 nm to 210 nm, such as from 210 nm to 220 nm, such as from 220 nm to 230 nm, such as from 230 nm to 240 nm, such as from 240 nm to 250 nm, such as from 250 nm to 260 nm, such as from 260 nm to 270 nm, such as from 270 nm to 280 nm, such as from 280 nm to 290 nm, such as from 290 nm to 300 nm, such as from 300 nm to 310 nm, such as from 310 nm to 320 nm, such as from 320 nm to 330 nm, such as from 330 nm to 340 nm, such as from 340 nm to 350 nm, such as from 350 nm to 360 nm, such as from 360 nm to 370 nm, such as from 370 nm to 380 nm, such as from 380 nm to 390 nm, such as from 390 nm to 400 nm, such as from 400 nm to 410 nm, such as from 410 nm to 420 nm, such as from 420 nm to 430 nm, such as from 430 nm to 440 nm, such as from 440 nm to 450 nm, such as from 450 nm to 460 nm, such as from 460 nm to 470 nm, such as from 470 nm to 480 nm, such as from 480 nm to 490 nm, such as from 490 nm to 500 nm, such as from 500 nm to 510 nm, such as from 510 nm to 520 nm, such as from 520 nm to 530 nm, such as from 530 nm to 540 nm, such as from 540 nm to 550 nm, such as from 550 nm to 560 nm, such as from 560 nm to 570 nm, such as from 570 nm to 580 nm, such as from 580 nm to 590 nm, such as from 590 nm to 600 nm, such as from 600 nm to 610 nm, such as from 610 nm to 620 nm, such as from 620 nm to 630 nm, such as from 630 nm to 640 nm, such as from 640 nm to 650 nm, such as from 650 nm to 660 nm, such as from 660 nm to 670 nm, such as from 670 nm to 680 nm, such as from 680 nm to 690 nm, such as from 690 nm to 700 nm, such as from 700 nm to 710 nm, such as from 710 nm to 720 nm, such as from 720 nm to 730 nm, such as from 730 nm to 740 nm, such as from 740 nm to 750 nm, such as from 750 nm to 760 nm, such as from 760 nm to 770 nm, such as from 770 nm to 780 nm, such as from 780 nm to 790 nm, such as from 790 nm to 800 nm, such as from 800 nm to 810 nm, such as from 810 nm to 820 nm, such as from 820 nm to 830 nm, such as from 830 nm to 840 nm, such as from 840 nm to 850 nm, such as from 850 nm to 860 nm, such as from 860 nm to 870 nm, such as from 870 nm to 880 nm, such as from 880 nm to 890 nm, such as from 890 nm to 900 nm, such as from 900 nm to 910 nm, such as from 910 nm to 920 nm, such as from 920 nm to 930 nm,Case Ref. P319WO IPTector®such as from 930 nm to 940 nm, such as from 940 nm to 950 nm, such as from 950 nm to 960 nm, such as from 960 nm to 970 nm, such as from 970 nm to 980 nm, such as from 980 nm to 990 nm, such as from 990 nm to 1000 nm.

[0169] In some embodiments, the hydrodynamic diameter is determined at a concentration of 5.56 mM of the nanostructure and a temperature of 298 K, following an annealing process.

[0170] In some embodiments, the hydrodynamic diameter (dH) and polydispersity index (PDI) of a nanostructure are determined by Dynamic Light Scattering (DLS) as set forth in the Examples. For example, samples in an H2O / 5% EtOH solution undergo an annealing process and are measured at 298 K, with the intensity distribution maxima used to determine dH.

[0171] In some embodiments, the hydrodynamic diameter (dH) is determined by Dynamic Light Scattering in a 1:19 EtOH: H2O solution at a concentration of less than 6 mM and a temperature of 298 K, following an annealing process.

[0172] In some embodiments, the nanostructure is characterized by having a polydispersity index (PDI) of less than 0.1, such as from 0.001 to 0.09, such as from 0.001 to 0.005, such as from 0.005 to 0.01, such as from 0.01 to 0.02, such as from 0.02 to 0.03, such as from 0.03 to 0.04, such as from 0.04 to 0.05, such as from 0.05 to 0.06, such as from 0.06 to 0.07, such as from 0.07 to 0.08, such as from 0.08 to 0.09.

[0173] In some embodiments, the nanostructure is characterized by having a Zeta potential of from -20 to -100 mV, such as from -100 to -90 mV, such as from -90 to -80 mV, such as from -80 to -70 mV, such as from -70 to -60 mV, such as from -60 to -50 mV, such as from -50 to -40 mV, such as from -40 to -30 mV, such as from -30 to -20 mV.

[0174] In some embodiments, the nanostructure is characterized by having a Zeta potential of from -40 to -90 mV.

[0175] In some embodiments, the Zeta potential of a nanostructure specified herein is determined as set forth in the Examples. For example, samples in an H2O / 5% EtOH solution undergo an annealing process and are measured at 298 K.

[0176] In some embodiments, the nanostructure is characterized by having a Critical Aggregation Concentration (CAC) of from 1 nM to 10 mM or above, for example from 1 nM to 10 mM, such as from 1 nM to 10 nM, such as from 10 nM to 50 nM, such as from 50 nM to 100 nM, such as from 100 nM to 500 nM, such as from 500 nM to 1.0 pM, such as from 1.0 pM to 5.0 pM, such as from 5.0 pM to 10 pM, such as from 10 pM to 50 pM, such as from 50 pM to 100 pM, such as from 100 pM to 200 pM, such as from 200 pM to 300 pM, such as from 300 pM to 400 pM, such as from 400 pM to 500 pM, such as from 500 pM to 600 pM, such as from 600 pM to 700 pM, such as from 700 pM to 800 pM, such as from 800 pM to 900 pM, such as from 900 pM to 1.0 mM, such as from 1.0 mM to 1.5Case Ref. P319WO IPTector®mM, such as from 1.5 mM to 2.0 mM, such as from 2.0 mM to 2.5 mM, such as from 2.5 mM to 3.0 mM, such as from 3.0 mM to 3.5 mM, such as from 3.5 mM to 4.0 mM, such as from 4.0 mM to 4.5 mM, such as from 4.5 mM to 5.0 mM, such as from 5.0 mM to 5.5 mM, such as from 5.5 mM to 6.0 mM, such as from 6.0 mM to 6.5 mM, such as from 6.5 mM to 7.0 mM, such as from 7.0 mM to 7.5 mM, such as from 7.5 mM to 8.0 mM, such as from 8.0 mM to 8.5 mM, such as from 8.5 mM to 9.0 mM, such as from 9.0 mM to 9.5 mM, such as from 9.5 mM to 10.0 mM.

[0177] In some embodiments, the nanostructure is characterized by having a Critical Aggregation Concentration (CAC) of from 0.3 mM to 10 mM or above, for example from 0.5 mM to 10 mM, such as from 0.5 mM to 1.0 mM, such as from 1.0 mM to 1.5 mM, such as from 1.5 mM to 2.0 mM, such as from 2.0 mM to 2.5 mM, such as from 2.5 mM to 3.0 mM, such as from 3.0 mM to 3.5 mM, such as from 3.5 mM to 4.0 mM, such as from 4.0 mM to 4.5 mM, such as from 4.5 mM to 5.0 mM, such as from 5.0 mM to 5.5 mM, such as from 5.5 mM to 6.0 mM, such as from 6.0 mM to 6.5 mM, such as from 6.5 mM to 7.0 mM, such as from 7.0 mM to 7.5 mM, such as from 7.5 mM to 8.0 mM, such as from 8.0 mM to 8.5 mM, such as from 8.5 mM to 9.0 mM, such as from 9.0 mM to 9.5 mM, such as from 9.5 mM to 10.0 mM.

[0178] In some embodiments, the nanostructure the nanostructure is characterized by having a surface tension (ST) at the Critical Aggregation Concentration (CAC) of from 30 to 55 mNm-1, such as from 30 to 32.5 mN-m-1, such as from 32.5 to 35 mN-m-1, such as from 35 to 37.5 mN-m-1, such as from 37.5 to 40 mN-m-1, such as from 40 to 42.5 mN-m-1, such as from 42.5 to 45 mN-m-1, such as from 45 to 47.5 mN-m-1, such as from 47.5 to 50 mN-m-1, such as from 50 to 52.5 mN-m-1, such as from 52.5 to 55 mN-m-1.

[0179] In some embodiments, the nanostructure the nanostructure is characterized by having a surface tension (ST) at the Critical Aggregation Concentration (CAC) of from 40 to 50 mNm-1.

[0180] In some embodiments, the nanostructure is characterized by having a Kdimof from 2.0 to 9000.0 M-1, such as from 2.0 to 10.0 M-1, such as from 10.0 to 50.0 M-1, such as from 50.0 to 100.0 M-1, such as from 100.0 to 200.0 M-1, such as from 200.0 to 300.0 M-1, such as from 300.0 to 400.0 M-1, such as from 400.0 to 500.0 M-1, such as from 500.0 to 600.0 M-1, such as from 600.0 to 700.0 M-1, such as from 700.0 to 800.0 M-1, such as from 800.0 to 900.0 M-1, such as from 900.0 to 1000.0 M-1, such as from 1000.0 to 2000.0 M-1, such as from 2000.0 to 3000.0 M-1, such as from 3000.0 to 4000.0 M-1, such as from 4000.0 to 5000.0 M-1, such as from 5000.0 to 6000.0 M-1, such as from 6000.0 to 7000.0 M-1, such as from 7000.0 to 8000.0 M-1, such as from 8000.0 to 9000.0 M-1.

[0181] In some embodiments, the nanostructure is characterized by having a Kdim of from 3.0 to 8000.0 M-1.

[0182] In some embodiments, the self-association constant (Kdim) is determined as set forth in theCase Ref. P319WO IPTector®Examples. For example, a solution of the compound undergoes serial dilution in a DMSO-d60.5% H2O solution at 298 K, and changes in1H NMR resonance positions are recorded with respect to SSA concentration. The data is subsequently fitted to an equal K / dimerization self-associative binding isotherm model using a suitable software program, such as Bindfit v0.5.

[0183] In some embodiments, the Critical Aggregation Concentration (CAC) is determined in a 1:19 EtOH: H2O solution following an annealing process.

[0184] In some embodiments, the surface tension (ST) is determined at the Critical Aggregation Concentration (CAC) in a 1:19 EtOH: H2O solution following an annealing process.

[0185] In some embodiments, the compound is characterized by exhibiting a membrane adhesion factor (MAF) of 0.5 or less and a membrane permeation factor (PF) of 1.0 or greater, when measured by1H CPMG NMR spectroscopy against 1:1 PE:PG phospholipid vesicles.

[0186] In some embodiments, the nanostructure has a hydrodynamic diameter (dH) of less than 250 nm, for example from 10 nm to 249 nm, for example from 120 nm to 250 nm.

[0187] In some embodiments, the nanostructure is characterized by a Critical Aggregation Concentration (CAC) of less than 85 mM.

[0188] In some embodiments, the nanostructure is characterized by a surface tension (ST) at the Critical Aggregation Concentration (CAC) of less than 50 mN-m-1, for example wherein the surface tension (ST) at the Critical Aggregation Concentration (CAC) is less than 45 mN-m-1.

[0189] In some embodiments, the formation of pores and / or channels is evidenced by the induction of one or more ion transport events corresponding to a current of at least 0.1 x 10−11A, for example at least 5.00 x 10−11A when the nanostructure comprising a plurality of compounds defined herein is applied to a planar phospholipid bilayer membrane at a concentration of 1.0 mM or less with respect to the compound and at a holding voltage of +100 mV.

[0190] In some embodiments, the ion transport events correspond to a current of from 0.1 x 10-11A to 5.0 x 10“8A, for example wherein the ion transport events correspond to a current of from 1.00 x 10“10A to 1.00 x 10“8A.

[0191] In some embodiments, the ion transport events correspond to a current of from 1.00 x 10“10A to 1.00 x 10“9A.

[0192] In some embodiments, the therapeutic agent is selected from the group consisting of: cisplatin, olaparib, gemcitabine, paclitaxel, and mitomycin C.Pharmaceutical composition

[0193] In some embodiments, a pharmaceutical composition is provided comprising a compound as defined herein, or a nanostructure as defined herein, and one or more pharmaceutically acceptableCase Ref. P319WO IPTector®excipients.

[0194] In some embodiments, the pharmaceutical composition further comprises a solvent.Further therapeutic agents

[0195] In some embodiments, the compound, nanostructure, or pharmaceutical composition as defined herein further comprises a therapeutic agent. In some embodiments, methods disclosed herein involving the compound, nanostructure, or pharmaceutical composition may further comprise administration of a therapeutic agent, such as a therapeutic agent disclosed herein.

[0196] In some embodiments, the therapeutic agent is a small molecule.

[0197] In some embodiments, the therapeutic agent is selected from the group consisting of: cisplatin, olaparib, gemcitabine, mitomycin C, and paclitaxel.

[0198] In some embodiments, the therapeutic agent is selected from the group consisting of: olaparib and paclitaxel.

[0199] In some embodiments, the therapeutic agent is cisplatin used in combination with SSA30.Membranes

[0200] In some embodiments, a phospholipid membrane is provided comprising the compound as defined herein, or the nanostructure as defined herein.

[0201] In some embodiments, a phospholipid membrane is provided comprising an ion-channel, such as a dynamic ion-channel or non-permanent ion-channel comprising the nanostructure as defined herein.

[0202] In some embodiments, the phospholipid membrane comprises one or more of: phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidic acid (PA), and sphingomyelin (SM).

[0203] In some embodiments, the phospholipid membrane is a biological cell membrane, such as a mammalian cell membrane, for example comprising PC, PE, PS, and / or SM, and optionally cholesterol.

[0204] In some embodiments, the phospholipid membrane comprises phospholipids with saturated or unsaturated fatty acid chains, such as palmitoyl, stearoyl, oleoyl, linoleoyl, arachidonoyl, or docosahexaenoyl (DHA) moieties.

[0205] In some embodiments, the phospholipid membrane comprises naturally occurring or synthetic phospholipids, such as l,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), l,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and l,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).Case Ref. P319WO IPTector®

[0206] In some embodiments, the membrane comprises (i) phosphatidylglycerol (PG); (ii) phosphatidylethanolamine (PE): PG 3:1 or; (iii) PE: PG 1:1.In some embodiments, the nanostructure is selected for lysing the membrane based on the constituents of the membrane. This is supported by at least Example 8.

[0207] In one embodiment, a nanostructure or compound is provided having an EC50towards a membrane comprising PG of from 0.0037 mM to 0.9913 mM, or 0.9913 mM or less. In some embodiments, the EC50is determined after 1 min, 5 min, 10 min, 15 min or 20 min after the nanostructure or compound is contacted with the membrane. The protocol and how to determine EC5o are described in more detail in Example 8.

[0208] In one embodiment, a nanostructure or compound is provided having an EC50towards a membrane comprising PE and PG, such as PE: PG in a ratio of 3:1, of from 0.0017 mM to 0.5529 mM, such as 0.5529 mM or less.

[0209] In one embodiment, a nanostructure or compound is provided having an EC50towards a membrane comprising PE and PG, such as PE: PG having a ratio of 1:1, of from 0.0015 mM to 0.9010 mM, such as 0.9010 mM or less.

[0210] In some embodiments, the membrane has been coated, penetrated, and / or disrupted by the nanostructure.

[0211] In some embodiments, the membrane is naturally occurring or synthetic.

[0212] In some embodiments, the membrane is provided wherein the nanostructure enables cation transport across the phospholipid membrane, for example wherein the cation is a sodium ion or a potassium ion.

[0213] In some embodiments, the membrane is provided wherein the nanostructure enables exchange of two different ions or molecules across the membrane in opposite directions.

[0214] In some embodiments, the membrane further comprises an anionophore.

[0215] Evidence to support the potential for SSAs and their nanostructures disclosed herein to form ion channel or pore-like structures was obtained via planar bilayer patch clamp experiment, using a semi-automated port-a-patch system as detailed in the present Examples, keeping experimental conditions as similar as possible to those used within the vesicle transport assays. Some differentiation between the methodologies was necessary to retain planar phospholipid bilayer stability.

[0216] These experiments show a variety of membrane ion transport events to occur, which have been first categorised by event type, defined by both current magnitude and time spent in that event state.

[0217] In some embodiments, a method of increasing or decreasing membrane rigidity is provided comprising contacting a nanostructure or compound as defined herein with a membrane, optionallyCase Ref. P319WO IPTector®a membrane as defined herein.

[0218] In some embodiments, a method of increasing membrane rigidity is provided, comprising contacting a nanostructure or compound as defined herein with a membrane, wherein Z of the compound of formula (I) is selected from the group consisting of a: a squaramide, and a thiourea. In some embodiments, a method is provided comprising a nanostructure comprising a plurality of different compounds, in some embodiments wherein the compounds comprise a different Z, such as a squaramide in one compound and a thiourea in another compound.Cells

[0219] In some embodiments, cell comprising the membrane as defined herein is provided.Treatment

[0220] In some embodiments, method for treatment of a pathological condition in a subject, the method comprising administering a compound, a nanostructure, or a pharmaceutical composition to the subject.

[0221] In some embodiments, the compound is as specified in any one of the paragraphs herein, the nanostructure is as specified in any one of the paragraphs herein, or the pharmaceutical composition is as specified in any one of the paragraphs herein.

[0222] In some embodiments, the pathological condition is a disease.

[0223] In some embodiments, the method further comprises administering, simultaneously or sequentially, a therapeutic agent to the subject.

[0224] In some embodiments of the method, the ratio of the therapeutic agent to the compound is from 1:5 to 1:20.

[0225] In some embodiments, the compound, nanostructure or pharmaceutical composition is administered to the subject orally, intraveneously, or intravesicularly.

[0226] In some embodiments, a compound, a nanostructure, or a pharmaceutical composition is provided for use in the treatment of a pathological condition in a subject, the use comprising administering the compound, the nanostructure, or the pharmaceutical composition to the subject.Cancer

[0227] In some embodiments, the method is provided wherein the pathological condition is a cancer. In some embodiments, the cancer is selected from the group consisting of: bladder cancer, and ovarian cancer.

[0228] In some embodiments, a method for treatment of cancer is provided comprisingCase Ref. P319WO IPTector®administering a combination comprising, separately or together, i) the compound, the nanostructure, or the pharmaceutical composition and ii) a therapeutic agent selected from the group consisting of: cisplatin, olaparib, gemcitabine, mitomycin C, and paclitaxel, to a subject having cancer. In some embodiments, the subject has bladder cancer and / or ovarian cancer.

[0229] In some embodiments, the compound, the nanostructure, or the pharmaceutical composition comprises compound (SSA48):

[0230] In some embodiments, for the treatment of cancer, the compound, the nanostructure, or the pharmaceutical composition comprises a compound selected from the group consisting of:Case Ref. P319WO IPTector®

[0231] In some embodiments, the compound, the nanostructure, or the pharmaceutical composition comprises compound (SSA30):

[0232] In some embodiments, the compound is SSA3. In some embodiments, the compound is SSA38.

[0233] In some embodiments, the cancer is sensitive or resistant cancer. In some embodiments, the cancer is sensitive or resistant ovarian cancer.

[0234] In some embodiments, the cancer is cisplatin sensitive cancer. In some embodiments, the cancer is cisplatin resistant cancer.

[0235] In some embodiments, a method of treating a cancer, such as bladder cancer is provided, wherein the method comprises administration of the compound, the nanostructure, or the pharmaceutical composition disclosed herein, wherein the compound is SSA3, and the treatment further comprises administration of mitomycin C.

[0236] In some embodiments, the treatment of bladder cancer comprises administration of a combination of SSA48 and olaparib.

[0237] In some embodiments, the treatment of bladder cancer comprises administration of a compound as disclosed herein and a therapeutic agent disclosed herein in a predefined ratio between a) therapeutic agent and b) the compound. In some embodiments, the predefined ratio is (of a:b): 1:1, 1:2.5, 1:5, or 1:10. In some embodiments, the predefined ratio is 1:1, 1:2.5, 1:5, or 1:10 and b is SSA48 and a is olaparib. In some embodiments, b is SSA48 and a is olaparib, wherein the ratio is at least 1:5 such as from 1:5 to 1:20, such as from 1:10 to 1:20.

[0238] In some embodiments, a is olaparib and b is SSA38, wherein the ratio is at least 1:5, such as 1:5 or 1:10, such as from 1:5 to 1:20, such as from 1:10 to 1:20.

[0239] In some embodiments, the subject having bladder cancer has cells with a histological type, mutation profile, and or aggressiveness in accordance with the cell line characteristics of: UM-UC-3, T24, 5637, and / or RT-4.

[0240] In some embodiments, the subject having ovarian cancer has cells with a histological type, mutation profile, and or aggressiveness in accordance with the cell line characteristics of: A2780 (cisplatin sensitive), A2780 CisR (cisplatin resistant subline), PE01 (cisplatin sensitive), PE04 (cisplatin resistant), and / or RPE-1 (non-cancerous hTERT human retinal pigment epithelial cell line).Case Ref. P319WO IPTector®

[0241] In some embodiments, the anticancer activity (e.g., GI50or IC50) is determined by a cell viability assay, such as the sulforhodamine B (SRB) assay, as set forth in the Examples. For example, cancer cells are seeded in 96-well plates, cultured, and then treated with a range of concentrations of the compound for a period of time, such as 96 hours. The cells are subsequently fixed with an agent such as trichloroacetic acid (TCA), stained with SRB dye, and the absorbance is measured to determine the concentration of the compound required to inhibit cell growth by 50% relative to untreated cells.Antimicrobial

[0242] In some embodiments, the method is provided wherein the pathological condition is a microbial infection.

[0243] In some embodiments, the microbial infection has resulted from a prokaryotic microorganism, archaea, or a eukaryotic microorganism.

[0244] In some embodiments, a method is provided for the treatment of a microbial infection which has resulted from one or more of Pseudomonas aeruginosa, Acinetobacter baumannii, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, and / or Enterococcus faecium.

[0245] In some embodiments, the microbial infection has resulted from a gram-positive bacterium or a gram-negative bacterium.

[0246] In some embodiments, the gram-positive bacterium is Staphylococcus aureus (S. Aureus), for example methicillin resistant S. aureus (MRSA).

[0247] In some embodiments, the gram-negative bacterium is Escherichia coli (E. coli).Biofilm application

[0248] In some embodiments, a method is provided for inhibiting biofilm on a surface of an object, the method comprising administering a compound or a nanostructure to the surface of the object.

[0249] In some embodiments, the method is provided wherein the compound is as defined herein, or the nanostructure is as defined herein.

[0250] In some embodiments, the antimicrobial activity (e.g., Minimum Inhibitory Concentration, MIC) is determined as set forth in the Examples. For example, a standardized inoculum of a microbial strain, such as S. aureus, is added to wells of a microtiter plate containing a serial dilution of the compound. The plates are incubated for a period of time, such as 20 hours, at an appropriate temperature, such as 37 °C, after which the MIC is determined as the lowest concentration of the compound at which no visible microbial growth is observed.Case Ref. P319WO IPTector®Forming nanostructures

[0251] In some embodiments, the present disclosure provides a process for preparing a nanostructure comprising a plurality of compounds each independently defined as specified herein, the process comprising:a) providing a plurality of compounds each independently defined as specified herein;b) subjecting the compounds to a liquid medium, such as an aqueous medium;c) thereby obtaining the nanostructure.

[0252] In some embodiments, a method is provided for synthesis of a compound as defined herein, such as a compound where Z comprises a urea, the method comprising: a) mixing an optionally protected amino acid with 1-isocyanato-4-(trifluoromethyl)benzene in a solvent, such as pyridine, to provide an intermediate, followed by b) deprotection of the protecting group, e.g. if the protecting group is a tert-butyl group, deprotection can be provided using zinc bromide in dichloromethane followed by addition of tetrabutylammonium (TBA) hydroxide in a polar protic solvent, such as methanol. Variations in this synthesis depend on the desired target compound. The present application provides examples of compound syntheses in Example 5.Methods of selecting relevant compounds

[0253] In some embodiments, method of selecting a compound capable of or suitable for forming pores and / or channels in a bilayer membrane, for example a phospholipid bilayer membrane, such as a biological cell membrane of a cell, the method comprising one or more or all of:a) Identifying a compound capable of complimentary interacting with phospholipid groups of a biological membrane of a cell;b) Identifying a compound engaging in self-associative hydrogen bonding events;c) Selecting a compound as defined herein;d) Subjecting a compound to patch clamp analysis to confirm presence of formation of pores and / or channels in the biological cell membrane,thereby selecting the compound suitable for or capable of forming pores and / or channels in the bilayer membrane.

[0254] In some embodiments, method of selecting a compound capable of or suitable for forming pores and / or channels in a bilayer membrane, for example a phospholipid bilayer membrane, such as a biological cell membrane of a cell, the method comprising:a) Subjecting a compound to patch clamp analysis to confirm presence of formation of pores and / or channels in the biological cell membrane,thereby selecting the compound suitable for or capable of forming pores and / or channels inCase Ref. P319WO IPTector®the bilayer membrane.

[0255] In some embodiments, the method is performed in vitro, ex vivo, or in vivo.

[0256] In some embodiments, the patch clamp analysis comprises subjecting one or more concentrations of a compound, for example a compound as defined herein, in one or more experiments to patch clamp analysis, applying an external voltage and after a course of time categorizing time spent in each of a number of event states.

[0257] The methodology of performing the patch clamp analysis for the present method is specified in detail in the Examples and displayed in the Figures.

[0258] In some embodiments, the patch clamp analysis comprises subjecting one or more concentrations of a compound to a membrane as specified herein, for example a bilayer membrane which is a phospholipid membrane.

[0259] In some embodiments, the number of event states are categorised as follows (current, A) 0: -00A < Event 0 < 5.00e-ll A, 1: 5.00e-ll A < Event 1 < 5.00e-10 A, 2: 5.00e-10 A < Event 2 < 4.90e-08 A, 3: 4.90e-08 A < Event 3 < °°.

[0260] In some embodiments, the external voltage is from +70 mV to +200 mv, for example +100 mV.

[0261] In some embodiments, the course of time is from 400 seconds to 2000 seconds, such as from 400 to 500 seconds, such as from 500 to 600 seconds, such as from 600 to 700 seconds, such as from 700 to 800 seconds, such as from 800 to 900 seconds, such as from 900 to 1000 seconds, such as from 1000 to 1100 seconds, such as from 1100 to 1200 seconds, such as from 1200 to 1300 seconds, such as from 1300 to 1400 seconds, such as from 1400 to 1500 seconds, such as from 1500 to 1600 seconds, such as from 1600 to 1700 seconds, such as from 1700 to 1800 seconds, such as from 1800 to 1900 seconds, such as from 1900 to 2000 seconds.

[0262] In some embodiments, the course of time is approximately 1000 seconds.

[0263] In some embodiments, the method is provided wherein a) detecting one or more increases in current over time and / or b) detecting at least 10% presence, such as at least 20% presence, such as at least 30% presence, such as at least 40% presence in more than one event states over the course of time confirms the compound being capable of or suitable for forming pores and / or channels in the biological cell membrane.Method of forming pores and / or channels in a membrane

[0264] In some embodiments, a method is provided of forming pores and / or channels in a membrane, such as a biological cell membrane of a cell using a nanostructure as defined herein, the method comprising contacting the nanostructure with the biological cell membrane of the cell in a liquid medium, for example an aqueous medium, thereby allowing the nanostructure to adhere toCase Ref. P319WO IPTector®and permeate the membrane, such as the biological cell membrane, forming pores and / or channels.

[0265] The mechanism is illustrated on Figure 2.

[0266] In some embodiments, a method is provided, for lysis of a vesicle comprising a membrane or a cell comprising a membrane, optionally a membrane as defined herein, said method comprising the steps of contacting the nanostructure as defined herein with the membrane. In some embodiments, the method further comprises one or more of the functional aspects described herein with respect to the interaction between the nanostructure and the membrane, c.f. " Function of nanostructures".

[0267] In some embodiments, the nanostructure or compound as defined herein is capable of lysing a vesicle and / or a cell.Method of delivering agent to cell

[0268] In some embodiments, a method is provided of using a compound as defined herein to deliver a therapeutic agent to a biological cell, comprising the steps:a) mixing the compound with the therapeutic agent in a liquid medium;b) forming a nanostructure comprising the compound and the therapeutic agent;c) delivering the nanostructure to a mammal comprising the biological cell, andd) allowing the nanostructure to adhere to and penetrate the biological cell.

[0269] In some embodiments, the biological cell is a pathogenic cell, such as a cancer cell or a microbial cell.

[0270] In some embodiments, the nanostructure forms dynamic pores and / or channels. Dynamic in this context means that the pore and / or channels are reversibly formed and that in some embodiments, the membrane with repair itself after contact with the nanostructure.Items1. A compound of formula (I) is provided,X-Z-Y (I),a tautomer, or a pharmaceutically acceptable salt thereof, whereinX, Y, and Z are intramolecularly connected, wherein X is a hydrophobic group, Z is a hydrogen bond donor and acceptor group, and Y is a hydrophilic hydrogen bond acceptor group.2. The compound of item 1, wherein the compound is capable of self-associating in a liquid medium.3. The compound of any one of the preceding items, wherein the compound is an amphiphile.Case Ref. P319WO IPTector®4. The compound of any one of the preceding items, wherein the compound is a self-associating amphiphile (SSA).5. The compound of any one of the preceding items, wherein the compound in a polar organic solvent, such as DMSO-dg 0.5 % H2O, exhibits a Kdimof from 15 to 140 M1, such as from 45 to 135 M1.6. The compound of any one of the preceding items, wherein the compound in a polar organic solvent, such as DMSO-d6(optionally with trace water), exhibits a dHof from 0.8 to 2.0 nm, such as from 1.4 to 1.9 nm.7. The compound of any one of the preceding items, wherein the compound exhibits an electrostatic surface potential energy maximum (Emax) of from -20 to -60 kJ. mol1, such as from -46 to -50 kJ·mol-1.8. The compound of any one of the preceding items, wherein the compound exhibits an electrostatic surface potential energy minimum (Emin) of from -650 to -800 kJ. mol1, such as from -700 to -725 kJ·mol-1.9. The compound of any one of the preceding items, wherein the compound exhibits an electrostatic surface potential energy minimum (Emin) as calculated by Spartan '24 of from 0 kJ-mol-1to -250 kJ-mol-1.10. The compound of any one of the preceding items, wherein the compound exhibits an electrostatic surface potential energy maximum (Emax) as calculated by Spartan '24 of from - 650 kJ-mol-1to -1050 kJ-mol-1.11. The compound of any one of the preceding items, wherein the compound has a cLogP value as calculated by SwissADME of less than 4.12. The compound of any one of the preceding items, wherein the compound in a polar organic solvent exhibits a dimerization constant (Kdim) of less than 3750 M-1.Case Ref. P319WO IPTector®13. The compound of any one of the preceding items, wherein the compound has a logP of from 2.50 to 5.50, for example from 3.0 to 4.50.14. The compound of any one of the preceding items, wherein the compound has increased capability of self-assembly.15. The compound of any one of the preceding items, wherein the compound is biologically active.16. The compound of any one of the preceding items, wherein the compound is biologically inactive.17. The compound of any preceding item, wherein the biological activity is selected from the group consisting of: anti-cancer activity, anti-microbial activity, and anti-biofilm activity.18. The compound of any preceding item, wherein the compound has an MIC against a microbial strain of less than 2.0 mM, such as from 5 pM to 1.5 mM.19. The compound of any preceding item, wherein the microbial strain is of a prokaryotic microorganism, archaea, or a eukaryotic microorganism.20. The compound of any preceding item, wherein the microbial strain is of a gram-positive bacterium or a gram-negative bacterium.21. The compound of any preceding item, wherein the gram-positive bacterium is Staphylococcus aureus (S. Aureus), for example methicillin resistant S. aureus (MRSA).22. The compound of any preceding item, wherein the gram-negative bacterium is is Escherichia coli (E. coli), Pseudomonas aeruginosa, or Acinetobacter baumannii.23. A compound of Formula (Al):R5R6R7R3Formula (Al),Case Ref. P319WO IPTector®wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selected from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;m is any integer, such as from 1 to 6;n is any integer, such as from 1 to 6;X is selected from the group consisting of: S, O, NH, NH2+, NR10, NHR10+, and SR10+; and wherein each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10is independently selected from the group consisting of: hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, such as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino, nitro, cyano, carboxyl, sulfonyl, phosphonyl, or fused with another one of R1, R2, R3, R4, R5, R6, R7, R8, R9, or R10of another compound of Formula (A1) forming a dimer between two compounds of Formula (A1); and wherein a pair of R1and R2, R2and R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of R1and R2, R2and R3, R3and R4, or R4and R5is bound in formula A1.24. The compound of any one of items 1-22, wherein the compound is of Formula (A3):Formula (A3);wherein each occurrence of Ri is independently selected from the group consisting of a lipophilic substituent and an electron-withdrawing substituent;wherein n is 1, 2, or 3,Case Ref. P319WO IPTector®for example wherein the lipophilic substituent is selected from the group consisting of C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C12 cycloalkyl, aryl, a non-aromatic ring system, a haloalkyl, such as C1-C24 haloalkyl, such as CF3, and SF5;for example wherein the electron-withdrawing substituent is selected from the group consisting of CF3, SF5, cyano, and halogen;wherein X is selected from the group consisting of O, S, guanidino, optionally substituted guanidinium, and optionally substituted thiouronium;wherein R3 is selected from the group consisting of hydrogen, C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, an aromatic ring system, and a non-aromatic ring system;wherein Y is selected from the group consisting of a phosphate, sulfate, and carboxylate; and wherein A is selected from the group consisting of a proton, and a carbon-containing cation.25. The compound of any one of items 1-22, wherein the compound is of Formula (A3):H H H H Formula (Cl);wherein R2 is selected from the group consisting of a lipophilic substituent and an electronwithdrawing substituent;wherein n is 1, 2, or 3,for example wherein the lipophilic substituent is selected from the group consisting of C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C12 cycloalkyl, aryl, a non-aromatic ring system, a haloalkyl, such as C1-C24 haloalkyl, such as CF3, and SF5;for example wherein the electron-withdrawing substituent is selected from the group consisting of CF3, SF5, cyano, and halogen;wherein X is selected from the group consisting of O, S, guanidino, optionally substituted guanidinium, and optionally substituted thiouronium;wherein R3 is selected from the group consisting of hydrogen, C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, an aromatic ring system, and a non-aromatic ring system;wherein Y is selected from the group consisting of a phosphate, sulfate, and carboxylate; and wherein A is selected from the group consisting of a proton, and a carbon-containing cation.26. The compound of item 25, wherein R2is selected from the group consisting of: C1-C24alkyl, C2-C24alkenyl, C2-C24alkynyl, C3-C14cycloalkyl, aryl, heteroaryl, C3-C14heterocyclyl, C1-C10haloalkyl, halogen, cyano, CF3, and SF5;Case Ref. P319WO IPTector®wherein each of said C1-C24alkyl, C2-C24alkenyl, C2-C24alkynyl, C3-C14cycloalkyl, aryl, heteroaryl, and C3-C14heterocyclyl groups is optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, hydroxy, -Ce alkoxy, C1-C6haloalkoxy, amino, mono(C1-C6alkyl)amino, di(C1-C6alkyl)amino, cyano, CF3, SF5, -C(=O)ORa, -C(=O)NRaRb, -SOzRa, and -SOzNRaRb;wherein any C1-C24alkyl, C2-C24alkenyl, or C2-C24alkynyl group in R2is optionally interrupted by 1 or 2 linkages independently selected from -O-, -S-, -NRa-, -C(=O)-, -C(=O)O-, -OC(=O)-, - C(=O)NRa-, and -NRaC(=O)-;wherein Ra and Rb are independently selected from H and C1-C6alkyl.27. The compound of item 26, wherein Rzis selected from the group consisting of: CTC12alkyl, Cz- C12alkenyl, CZ-C1Zalkynyl, C3-C10cycloalkyl, aryl, heteroaryl, -Cs haloalkyl, CF3, SF5, and cyano; wherein each of said C C12alkyl, C2-C12alkenyl, C2-C12alkynyl, C3-C10cycloalkyl, aryl, and heteroaryl groups is optionally substituted with 1 or 2 substituents independently selected from halogen, cyano, CF3, SF5, C1-C4alkoxy, -C(=O)ORa, and -C(=O)NRaRb;wherein any C1-C12alkyl, C2-C12alkenyl, or C2-C12alkynyl group in R2is optionally interrupted by 1 linkage selected from -O-, -C(=O)O-, and -C(=O)NRa-;wherein C1-C12alkyl includes linear and branched alkyl.28. The compound of any one of items 1-22, wherein the compound is of Formula (A3):Formula (A3);wherein each occurrence of Ri is independently selected from the group consisting of a lipophilic substituent and an electron-withdrawing substituent;wherein n is 1, 2, or 3,for example wherein the lipophilic substituent is selected from the group consisting of C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C12 cycloalkyl, aryl, a non-aromatic ring system, a haloalkyl, such as C1-C24 haloalkyl, such as CF3, and SF5;for example wherein the electron-withdrawing substituent is selected from the group consisting of CF3, SF5, cyano, and halogen;wherein X is selected from the group consisting of O, S, guanidino, optionally substituted guanidinium, and optionally substituted thiouronium;wherein R3 is selected from the group consisting of hydrogen, C1-C24 alkyl, C2-C24 alkenyl, C2-C24Case Ref. P319WO IPTector®alkynyl, an aromatic ring system, and a non-aromatic ring system;wherein Y is selected from the group consisting of a phosphate, sulfate, and carboxylate; and wherein A is selected from the group consisting of a proton, and a carbon-containing cation.29. The compound of any one of items 24-28, wherein A is selected from the group consisting of: a proton (H+), a substituted ammonium group, preferably tert-butyl ammonium (TBA), and a bioactive compound, for example a proton or TBA.30. The compound of any one of items 1-29, wherein when Y carries a negative charge, A is present in an amount that provides overall electroneutrality.31. The compound of any one of items 24, 28, and 29-30, wherein n is 1.32. The compound of any one of items 24, 28, 29-31, wherein each occurrence of Ri is independently selected from the group consisting of: C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C14 cycloalkyl, aryl, heteroaryl, C3-C14 heterocyclyl, C1-C10 haloalkyl, halogen, cyano, CF3, and SF5;wherein each of said C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C14 cycloalkyl, aryl, heteroaryl, and C3-C14 heterocyclyl groups is optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, hydroxy, C1-C6alkoxy, C1-C6haloalkoxy, amino, mono(C1-C6alkyl)amino, di(C1-C6alkyl)amino, cyano, CF3, SF5, -C(=O)ORa, -C(=O)NRaRb, -SO2Ra, and -SO2NRaRb;wherein any C1-C24 alkyl, C2-C24 alkenyl, or C2-C24 alkynyl group in Ri is optionally interrupted by 1 or 2 linkages independently selected from -O-, -S-, -NRa-, -C(=O)-, -C(=O)O-, -OC(=O)-, - C(=O)NRa-, and -NRaC(=O)-;wherein Raand Rbare independently selected from H and C1-C6alkyl.33. The compound of item 32, wherein each occurrence of Ri is independently selected from the group consisting of: C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C3-C10 cycloalkyl, aryl, heteroaryl, C1-C6haloalkyl, CF3, SF5, and cyano;wherein each of said C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C3-C10 cycloalkyl, aryl, and heteroaryl groups is optionally substituted with 1 or 2 substituents independently selected from halogen, cyano, CF3, SF5, C1C1-C4alkoxy, -C(=O)ORa, and -C(=O)NRaRb;wherein any C1-C12 alkyl, C2-C12 alkenyl, or C2-C12 alkynyl group in Ri is optionally interrupted byCase Ref. P319WO IPTector®1 linkage selected from -O-, -C(=O)O-, and -C(=O)NRa-.34. The compound of any one of items 24-32, wherein R3 is selected from the group consisting of:H, C1-C24 alkyl, C2-C24alkenyl, C2-C24alkynyl, C3-C14cycloalkyl, C6-C14aryl, C7-C20aryl(C1-C6)alkyl, 5- to 14-membered heteroaryl, and 3- to 14-membered non-aromatic heterocyclyl; whereinalkyl includes linear and branched alkyl;wherein each R3other than H is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen, hydroxy, oxo, cyano, nitro, amino, mono(C1-C6alkyl)amino, di(C1-C6alkyl)amino, C C6alkyl, -Ce haloalkyl, -Ce alkoxy, C1-C6haloalkoxy, -Ce alkylthio, CF3, SF5, carboxyl, -Ce alkoxycarbonyl, -Ce alkylcarbonyl, -C(=O)NH2, -Cf^jNHfCr C6alkyl), -C(=O)N(C1-C6alkyl )2, -SO2(C C6alkyl), -SO2NH2, -SO2NH(C1-C6alkyl), -SO2N(C1-C6alkyl)2, -P(=O)(OH)2, and -P(=O)(O(C C6alkyl))2;wherein any C1-C24alkyl, C2-C24alkenyl, or C2-C24alkynyl portion of R3is optionally interrupted by 1 or 2 linkages independently selected from -O-, -S-, -NH-, -N(C1-C6alkyl)-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH-, -NHC(=O)-, -C(=O)N(C1-C6alkyl)-, and -N(C1-C6alkyl)C(=O)-.35. The compound of item 34, wherein R3is selected from the group consisting of H, C1-C12alkyl, C2-C12alkenyl, C2-C12alkynyl, C3-C10cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, C7-C16aryl(C1-C4)alkyl, 5- to 10-membered heteroaryl, and 5- to 10-membered non-aromatic heterocyclyl; wherein C1-C12alkyl includes linear and branched alkyl, including isobutyl;wherein C7-C16aryl(C1-C4)alkyl includes benzyl, 1-naphthylmethyl, and 2-naphthylmethyl; wherein each R3group other than H is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen, cyano, CF3, SF5, -C4 alkyl, -C4 haloalkyl, C1-C4alkoxy, -C4 haloalkoxy, amino, mono(C1-C4alkyl)amino, di(CrC4alkyl)amino, - C(=0)0(C C4 alkyl), -C(=O)NH2, -C^OjNHfC^ alkyl), -SOzfC^ alkyl), and -SO2NH2;and wherein any alkyl, alkenyl, or alkynyl portion of R3is optionally interrupted by 1 linkage selected from -O-, -C(=O)O-, and -C(=O)NH-.36. The compound of item 35, wherein R3is selected from the group consisting of isobutyl, secbutyl, tert-butyl, benzyl, 1-naphthylmethyl, and 2-naphthylmethyl, each optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen, -C4 alkyl, C1-C4alkoxy, CF3, SF5, and cyano.37. The compound of any one of preceding items, wherein each occurrence of Ri is independentlyCase Ref. P319WO IPTector®selected from the group consisting of C1–C12alkyl, C2–C12alkenyl, C2–C12alkynyl, C3–C12cycloalkyl, aryl, heteroaryl, non-aromatic ring system, CF3, and SF5, cyano, nitro, halogen, C(=O)C1–C6alkyl, C(=O)O–C1–C6alkyl, C(=O)NH2, C(=O)NH(C1–C6alkyl), SO2–C1–C6alkyl, SO2NH2, and SO2NH(C1–C6alkyl).38. The compound of any one of preceding items, wherein X is selected from the group consisting of O, S, guanidino, optionally substituted guanidinium, and optionally substituted thiouronium.39. The compound of any one of preceding items, wherein the compound is of Formula (A4);Formula (A4),wherein R3 is isobutyl or benzyl.40. The compound of any one of preceding items, wherein the compound is of Formula (Bl):R2 Formula (Bl);wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selected from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;m is any integer, such as from 1 to 6;n is any integer, such as from 1 to 6; andwherein each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10is independently selected from the group consisting of: hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, suchCase Ref. P319WO IPTector®as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino, nitro, cyano, carboxyl, sulfonyl, phosphonyl, or fused with another one of R1, R2, R3, R4, R5, R6, R7, R8, R9, or R10of another compound of Formula (B1) forming a dimer between two compounds of Formula (Bl); and wherein a pair of R1and R2, R2and R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of R1and R2, R2and R3, R3and R4, or R4and R5is bound in formula B1.41. The compound of any one of preceding items, wherein the compound is of Formula (B2):AFormula (B2);wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selected from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;m is any integer, such as from 1 to 6;n is any integer, such as from 1 to 6; andwherein each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10is independently selected from the group consisting of: hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, such as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino,Case Ref. P319WO IPTector®nitro, cyano, carboxyl, sulfonyl, phosphonyl, or fused with another one of R1, R2, R3, R4, R5, R6, R7, R8, R9, or R10of another compound of Formula (B2) forming a dimer between two compounds of Formula (B2); and wherein a pair of R1and R2, R2and R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of R1and R2, R2and R3, R3and R4, or R4and R5is bound in formula B2.42. The compound of any one of the preceding items, wherein Y comprises a carboxylate anion.43. The compound of any one of the preceding items, wherein A comprises tetrabutylammonium (TBA).44. The compound of any one of the preceding items, wherein R6and / or R7are a hydrophobic group.45. The compound of any one of the preceding items, wherein one of R6and R7is a hydrophobic group and one is hydrogen.46. The compound of any one of the preceding items, wherein one of R6and R7is a branched or straight alkyl chain, such as isobutyl, or an aralkyl, such as benzyl.47. The compound of any one of the preceding items, wherein at least 2, such as at least 3, such as at least 4 of R1, R2, R3, R4, and R5is hydrogen.48. The compound of any one of the preceding items, wherein one of R1, R2, R3, R4, and R5is a trifluoroalkyl, such as trifluoromethyl.49. The compound of any one of the preceding items, wherein the compound is chiral.50. The compound of any one of the preceding items, wherein the compound is essentially a single enantiomer or a mixture of enantiomers, for example essentially a 1:1 mixture of enantiomers.51. The compound of any one of the preceding items, wherein the compound is of formula A2, A1, B1, or B2, and wherein one or more of R6, R7, R8, and R9are independently selected from H andCase Ref. P319WO IPTector® a hydrophobic group, wherein at least one of R6, R7, R8, and R9is a hydrophobic group.52. The compound of any one of the preceding items, wherein the compound is not any one of:Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®53. The compound of any one of the preceding items, wherein the compound is selected from the group consisting of:Case Ref. P319WO IPTector® L118 / D119 L120 / D121 L124 / D125Case Ref. P319WO IPTector®174175176Case Ref. P319WO IPTector®54. The compound of any one of the preceding items, wherein the compound is selected from the group consisting of:Case Ref. P319WO IPTector®stereoisomeric mixture thereof (SSA 72, 73, 56, 57, 74, 75).55. The compound of any one of the preceding items, wherein the compound is selected from the group consisting of:173Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®, a stereoisomer thereof, and a salt thereof, such as a pharmaceutically acceptable salt thereof.56. The compound of any one of the preceding items, wherein the compound is characterized by exhibiting a membrane adhesion factor (MAF) of 0.5 or less and a membrane permeation factor (PF) of 1.0 or greater, when measured by1H CPMG NMR spectroscopy against 1:1 PE:PG phospholipid vesicles.57. A nanostructure comprising a plurality of compounds each independently defined as in any one of items 1-56.58. The nanostructure of item 57, wherein the plurality of compounds self-associate in a liquid medium, such as an aqueous medium, thereby forming the nanostructure.59. The nanostructure of any of items 57-58, wherein the nanostructure is configured for or capable of interacting with a biological membrane and adhering to and / or penetrating the biological membrane to form pores and / or channels in the biological membrane.60. The nanostructure of item 59, wherein the adhering to and / or penetrating the biological membrane increases the susceptibility of the biological membrane to a therapeutic agent and / or increases the permeability of the therapeutic agent across the biological membrane when contacted with the therapeutic agent.61. The nanostructure of any of items 57-60, wherein the nanostructure is capable of or configured for acting as a supramolecular host to one or more phospholipid headgroups of a biological membrane acting as supramolecular guests.62. The nanostructure of any of items 57-61, wherein the nanostructure is capable of forming or forms a supramolecular host:guest complex with one or more phospholipid headgroups of a biological membrane when contacted with one or more phospholipid headgroups of a biologicalCase Ref. P319WO IPTector®membrane.63. The nanostructure of any of items 57-62, wherein the nanostructure is capable of or configured for absorbing or adsorbing a therapeutic agent, such as a small molecule.64. The nanostructure of any of items 57-63, wherein the nanostructure is capable of or configured for absorbing or adsorbing a therapeutic agent, such as a small molecule, and further capable of or configured for delivering the therapeutic agent to a biological cell.65. The nanostructure of any of items 57-59, wherein the plurality of compounds comprises the same or different compounds.66. The nanostructure of any of items 57-65, wherein the plurality of compounds is selected from the group consisting of:Case Ref. P319WO IPTector®10Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®o67. The nanostructure of any of items 57-66, wherein the plurality of compounds is selected from the group consisting of:stereoisomeric mixtures thereof.Case Ref. P319WO IPTector®68. The nanostructure of any of items 57-68, wherein the plurality of compounds is selected from the group consisting of:stereoisomeric mixtures thereof.69. The nanostructure of any one of items 57-68, comprising two different compounds.70. The nanostructure of any one of items 57-69, comprising a combination of compounds selected from the group consisting of:Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®71. The nanostructure of any of items 57-70, wherein the nanostructure comprises a mixture of enantiomers, for example in a ratio of from 30 to 70, such as from 35 to 65, such as from 40 to 60, such as for example approximately 50:50.72. The nanostructure of any of items 57-71, wherein the plurality of compounds comprises essentially a single enantiomer.73. The nanostructure of any of items 57-72, wherein the nanostructure is capable of aggregating into an essentially spherical form, or a gel fiber, or a combination thereof.74. The nanostructure of item 73, wherein the nanostructure is essentially spherical when in a liquid medium, such as an aqueous medium.75. The nanostructure of any one of items 57-74, wherein the nanostructure has a hydrodynamic diameter, dHof from 90 to 600 nm, such as from 90 nm to 100 nm, such as from 100 nm to 110 nm, such as from 110 nm to 120 nm, such as from 120 nm to 130 nm, such as from 130 nm to 140 nm, such as from 140 nm to 150 nm, such as from 150 nm to 160 nm, such as from 160 nm to 170 nm, such as from 170 nm to 180 nm, such as from 180 nm to 190 nm, such as from 190 nm to 200 nm, such as from 200 nm to 210 nm, such as from 210 nm to 220 nm, such as from 220 nm to 230 nm, such as from 230 nm to 240 nm, such as from 240 nm to 250 nm, such as from 250 nm to 260 nm, such as from 260 nm to 270 nm, such as from 270 nm to 280 nm, such as from 280 nm to 290 nm, such as from 290 nm to 300 nm, such as from 300 nm to 310 nm, such as from 310 nm to 320 nm, such as from 320 nm to 330 nm, such as from 330 nm to 340 nm, such as from 340 nm to 350 nm, such as from 350 nm to 360 nm, such as from 360 nm to 370 nm, such as from 370 nm to 380 nm, such as from 380 nm to 390 nm, such as from 390 nm to 400 nm, such as from 400 nm to 410 nm, such as from 410 nm to 420 nm, such as from 420 nm to 430 nm, such as from 430 nm to 440 nm, such as from 440 nm to 450 nm, such as from 450 nm to 460 nm, such as from 460 nm to 470 nm, such as from 470 nm to 480 nm, such as from 480 nm to 490 nm, such as from 490 nm to 500 nm, such as from 500 nm to 510 nm, suchCase Ref. P319WO IPTector®as from 510 nm to 520 nm, such as from 520 nm to 530 nm, such as from 530 nm to 540 nm, such as from 540 nm to 550 nm, such as from 550 nm to 560 nm, such as from 560 nm to 570 nm, such as from 570 nm to 580 nm, such as from 580 nm to 590 nm, such as from 590 nm to 600 nm, such as from 600 nm to 610 nm, such as from 610 nm to 620 nm, such as from 620 nm to 630 nm, such as from 630 nm to 640 nm, such as from 640 nm to 650 nm, such as from 650 nm to 660 nm, such as from 660 nm to 670 nm, such as from 670 nm to 680 nm, such as from 680 nm to 690 nm, such as from 690 nm to 700 nm, such as from 700 nm to 710 nm, such as from 710 nm to 720 nm, such as from 720 nm to 730 nm, such as from 730 nm to 740 nm, such as from 740 nm to 750 nm, such as from 750 nm to 760 nm, such as from 760 nm to 770 nm, such as from 770 nm to 780 nm, such as from 780 nm to 790 nm, such as from 790 nm to 800 nm, such as from 800 nm to 810 nm, such as from 810 nm to 820 nm, such as from 820 nm to 830 nm, such as from 830 nm to 840 nm, such as from 840 nm to 850 nm, such as from 850 nm to 860 nm, such as from 860 nm to 870 nm, such as from 870 nm to 880 nm, such as from 880 nm to 890 nm, such as from 890 nm to 900 nm, such as from 900 nm to 910 nm, such as from 910 nm to 920 nm, such as from 920 nm to 930 nm, such as from 930 nm to 940 nm, such as from 940 nm to 950 nm, such as from 950 nm to 960 nm, such as from 960 nm to 970 nm, such as from 970 nm to 980 nm, such as from 980 nm to 990 nm, such as from 990 nm to 1000 nm.76. The nanostructure of any one of items 57-75, wherein the nanostructure has a hydrodynamic diameter (dH) of less than 250 nm, for example from 10 nm to 249 nm, for example from 120 nm to 250 nm.77. The nanostructure of any one of items 57-76, wherein the nanostructure is characterized by having a polydispersity index (PDI) of less than 0.1, such as from 0.001 to 0.09, for example wherein the nanostructure has a polydispersity index (PDI) of less than 0.05.78. The nanostructure of any one of items 57-77, wherein the nanostructure is characterized by having a Zeta potential of from -20 to -100 mV, for example from -40 to -90 mV or from -70 mV to -30 mV.79. The nanostructure of any one of items 57-78, wherein the nanostructure is characterized by a Critical Aggregation Concentration (CAC) of less than 85 mM.80. The nanostructure of any one of items 57-79, wherein the nanostructure is characterized byCase Ref. P319WO IPTector®having a Critical Aggregation Concentration (CAC) of from 0.3 mM to 10 or above mM, for example from 0.5 mM to 10 mM.81. The nanostructure of any one of items 57-80, wherein the nanostructure is characterized by a surface tension (ST) at the Critical Aggregation Concentration (CAC) of less than 50 mN-m-1, for example wherein the surface tension (ST) at the Critical Aggregation Concentration (CAC) is less than 45 mN-m-1.82. The nanostructure of any one of items 57-81, wherein the nanostructure is characterized by having a surface tension (ST) at the Critical Aggregation Concentration (CAC) of from 30 to 55 mNm1, such as from 40 to 50 mNm-1.83. The nanostructure of any one of items 57-82, wherein the nanostructure is characterized by having a K of from 2.0 to 9000.0 M-1, such as from 3.0 to 8000.0 M-1.84. The nanostructure of any one of items 57-83, wherein the nanostructure is capable of or configured for forming pores and / or channels in a biological cell membrane, for example as evidenced by patch clamp analysis.85. The nanostructure of item 84, wherein the formation of pores and / or channels is evidenced by the induction of one or more ion transport events corresponding to a current of at least 0.1 x 10−11A, for example at least 5.00 x 10−11A when the nanostructure is applied to a planar phospholipid bilayer membrane at a concentration of 1.0 mM or less with respect to the compound and at a holding voltage of +100 mV.86. The nanostructure of item 84, wherein the ion transport events correspond to a current of from 0.1 x 10-11A to 5.0 x 10“8A, for example wherein the ion transport events correspond to a current of from 1.00 x 10“10A to 1.00 x 10“8A.87. The nanostructure of item 84, wherein the ion transport events correspond to a current of from 1.00 x IO’10A to 1.00 x 10“9A.88. A pharmaceutical composition comprising a compound as defined in any one of items 1-34, or a nanostructure as defined in any one of items 35-52, and one or more pharmaceuticallyCase Ref. P319WO IPTector®acceptable excipients.89. The pharmaceutical composition of item 88, wherein the composition further comprises a solvent.90. The compound, nanostructure, or pharmaceutical composition of any one of the preceding items, further comprising a therapeutic agent.91. The compound, nanostructure, or pharmaceutical composition according to item 90, wherein the therapeutic agent is a small molecule.92. The compound, nanostructure, or pharmaceutical composition according to any of items 90-91, wherein the therapeutic agent is selected from the group consisting of: cisplatin, olaparib, gemcitabine, paclitaxel, and mitomycin C.93. The compound, nanostructure, or pharmaceutical composition according to any of items 90-92, wherein the therapeutic agent is selected from the group consisting of: cisplatin, olaparib and paclitaxel.94. The compound, nanostructure, or pharmaceutical composition according to any of preceding items, further comprising a therapeutic agent selected from the group consisting of: olaparib and paclitaxel.95. A phospholipid membrane comprising the compound as defined in any one of items 1-56, or the nanostructure as defined in any one of items 57-84.96. The phospholipid membrane of item 95, wherein the membrane has been coated, penetrated, and / or disrupted by the nanostructure.97. The phospholipid membrane of any of items 95-96, wherein the membrane is naturally occurring or synthetic.98. The phospholipid membrane of any of items 95-97, wherein the nanostructure enables cation transport across the phospholipid membrane, for example wherein the cation is a sodium ionCase Ref. P319WO IPTector®or a potassium ion.99. The phospholipid membrane of any of items 95-97, wherein the nanostructure enables exchange of two different ions or molecules across the membrane in opposite directions.100. The phospholipid membrane of any of items 95-98, wherein the membrane further comprises an anionophore.101. A cell comprising the membrane as defined in any one of items 95-100.102. A method for treatment of a pathological condition in a subject, the method comprising administering a compound, a nanostructure, or a pharmaceutical composition to the subject.103. The method of item 72, wherein the compound is as defined in any one of items 1-54, the nanostructure is as defined in any one of items 57-75, or the pharmaceutical composition is as defined in item 88.104. The method of item 102, wherein the pathological condition is a disease.105. The method of any one of items 102-104, wherein the method further comprises administering, simultaneously or sequentially, a therapeutic agent to the subject.106. The method of item 105, wherein the ratio of the therapeutic agent to the compound is from 1:5 to 1:20.107. The method of any one of items 102-106, wherein the compound, nanostructure or pharmaceutical composition is administered to the subject orally, intraveneously, or intravesicularly.108. A compound, a nanostructure, or a pharmaceutical composition for use in the treatment of a pathological condition in a subject, the use comprising administering the compound, the nanostructure, or the pharmaceutical composition to the subject.109. The method of any one of items 102-107, wherein the pathological condition is a cancer.Case Ref. P319WO IPTector®110. The methof item 109, wherein the cancer is selected from the group consisting of: bladder cancer, and ovarian cancer.111. The method of any of items 109-110, wherein the compound, the nanostructure, or the pharmaceutical composition comprises compound (SSA48):ss,O’ TBA(SSA48).112. The method of any one of items 102-111, wherein the pathological condition is a microbial infection.113. The method of of item 112, wherein the microbial infection has resulted from a gram-positive bacterium or a gram-negative bacterium.114. The method of item 113, wherein the gram-positive bacterium is Staphylococcus aureus (S.Aureus), for example methicillin resistant S. aureus (MRSA).115. The method of item 113, wherein the gram-negative bacterium is Escherichia coli (E. coli).116. A method for inhibiting biofilm on a surface of an object, the method comprising administering a compound or a nanostructure to the surface of the object.117. The method of item 116, wherein the compound is defined as in any one of items 1-56, or the nanostructure is defined as in any one of items 57-84.118. A process for preparing a nanostructure comprising a plurality of compounds each independently defined as in any one of items 1-56, the process comprising:a. providing a plurality of compounds each independently defined as in any one of items 1-56; b. subjecting the compounds to a liquid medium, such as an aqueous medium;c. thereby obtaining the nanostructure.119. A method of selecting a compound capable of or suitable for forming pores and / or channels in a bilayer membrane, for example a phospholipid bilayer membrane, such as a biological cellCase Ref. P319WO IPTector®membrane of a cell, the method comprising one or more or all of:a) Identifying a compound capable of complimentary interacting with phospholipid groups of a biological membrane of a cell;b) Identifying a compound engaging in self-associative hydrogen bonding events;c) Selecting a compound as defined in any one of items 1-56;d) Subjecting a compound to patch clamp analysis to confirm presence of formation of pores and / or channels in the biological cell membrane,thereby selecting the compound suitable for or capable of forming pores and / or channels in the bilayer membrane.120. The method of item 119, wherein the method is performed in vitro, ex vivo, or in vivo.121. The method of any of items 119-120, wherein the patch clamp analysis comprises subjecting one or more concentrations of a compound, for example a compound as defined in any one of items 1-34, in one or more experiments to patch clamp analysis, applying an external voltage and after a course of time categorizing time spent in each of a number of event states.122. The method of any of items 119-121, wherein the patch clamp analysis comprises subjecting one or more concentrations of a compound to a bilayer membrane which is a phospholipid membrane.123. The method of any of items items 119-122, wherein the number of event states are categorised as follows (current, A) 0: -co A < Event 0 < 5.00e-ll A, 1: 5.00e-ll A < Event 1 < 5.00e-10 A, 2: 5.00e-10 A < Event 2 < 4.90e-08 A, 3: 4.90e-08 A < Event 3 < co.124. The method of any of items

[0419] 121-

[0419] 123, wherein the external voltage is from +70 mV to +200 mv, for example +100 mV.125. The method of any of items 121-124, wherein the course of time is from 400 seconds to 2000 seconds, such as approximately 1000 seconds.126. The method of any of items 121-125, further comprising measuring the change in current (A) over time and detecting one or more increases in current over time.Case Ref. P319WO IPTector®127. The method of any of items 121-126, wherein a) detecting one or more increases in current over time and / or b) detecting at least 10% presence, such as at least 20% presence, such as at least 30% presence, such as at least 40% presence in more than one event states over the course of time confirms the compound being capable of or suitable for forming pores and / or channels in the biological cell membrane.128. A method of forming pores and / or channels in a biological cell membrane of a cell using a nanostructure as defined in any one of items 57-84, the method comprising contacting the nanostructure with the biological cell membrane of the cell in a liquid medium, for example an aqueous medium, thereby allowing the nanostructure to adhere to and permeate the biological cell membrane forming pores and / or channels.129. A method of using a compound as defined in any one of items 1-56 to deliver a therapeutic agent to a biological cell, comprising the steps:a. mixing the compound with the therapeutic agent in a liquid medium;b. forming a nanostructure comprising the compound and the therapeutic agent;c. delivering the nanostructure to a mammal comprising the biological cell, andd. allowing the nanostructure to adhere to and penetrate the biological cell.130. The method of any of items 128-129, wherein the biological cell is a pathogenic cell, such as a cancer cell or a microbial cell.131. The method of any of items 128-130, wherein the nanostructure forms dynamic pores and / or channels.ExamplesExample 1 - General methods

[0271] A positive pressure of nitrogen and oven dried glassware were used for all reactions. All solvents and starting materials were purchased from known chemical suppliers or available stores and used without any further purification unless specifically stipulated. The NMR spectra were obtained using a Bruker AV2400 MHz or AVNEO 400 MHz spectrometer. The data was processed using TopSpin 4.1.4. software. NMR chemical shift values are reported in parts per million (ppm) and calibrated to the centre of the residual solvent peak set (s = singlet, br = broad, d = doublet, t = triplet, q = quartet,Case Ref. P319WO IPTector®m = multiplet). Tensiometry measurements were undertaken using the Biolin Scientific Theta Attension optical tensiometer. The data was processed using Biolin OneAttension software. A Hamilton (309) syringe was used for these measurements. The melting point for each compound was measured using Stuart SMP10 melting point apparatus. High resolution mass spectrometry was performed using a Bruker microTOF-Q mass spectrometer and spectra recorded and processed using Bruker's Compass Data Analysis software. Infrared spectra were obtained using Shimadzu I R-Affinity-1 model Infrared spectrometer. The data was analysed in wavenumbers (cm1) using IRsolution software. DLS and Zeta Potential studies were carried out using an Anton Paar Litesizer™ 500 and processed using Kalliope™ professional software.from diffusion rates obtained from1H NMR DOSY measurements using the Stokes-Einstein equation. The viscosity value used for the calculation was 0.00199 mPa (DMSO).Quantitative1H NMR (qNMR) studies:

[0272] A1H NMR spectrum was obtained with a delay (d1= 60 s) for compounds (112 mM) in DMSO-d6 / 1.0 % DCM or 5.56 mM in D2O / 5.0 % EtOH. Through comparative integration of the anionic and cationic component signals against the internal standard signals (DCM / EtOH), the proportion of these molecular components to become 'lost' from solution, through the adoption of solid-like characteristics can be calculated.Self-association constant calculation:

[0273] Self-association constants (Kdim) were determined using Bindfit v0.5. Here a solution of the compound underwent serial dilution in a DMSO-d60.5 % H2O solution at 298 K. Changes in 1H NMR resonance positions were then recorded with respect to SSA concentrationTensiometry studies:

[0274] All the samples were prepared in an H2O / 5 % EtOH solution. All samples underwent an annealing process where they were heated to approximately 313 K before being left to cool to RT, enabling each sample to reach a thermodynamic minimum. All samples were prepared through serial dilution of the most concentrated sample. Three surface tension measurements were obtained for each sample at a given concentration using the pendant drop method. The average values were then used to calculate the CAC.Mass spectrometry studies:

[0275] Approximately 1 mg of each compound was dissolved in 1 mL of MeOH. This solution was further diluted 100-fold before undergoing analysis where 10 pL of each sample was injected directly into a flow of 10 mM ammonium acetate in 95 % water (flow rate = 0.02 mL / min).Dynamic light scattering (DLS) studies:

[0276] All solvents used were filtered to remove any particulates that may interfere with the resultsCase Ref. P319WO IPTector®obtained. All samples underwent an annealing process, in which they were heated to 313 K before being left to cool to 298 K to allow each sample to reach a thermodynamic minimum. A series of 10 runs were recorded at 298 K. These results were then combined to determine the intencity distribution maxima. Studies were undertaken in a variety of solvents and concnetrations which include H2O / 5.0 % EtOH.Zeta potential studies:

[0277] All solvents used were filtered to remove any particulates that may interfere with the results obtained. All samples underwent an annealing process in which they were heated to 313 K before being left to cool to 298 K to allow each sample to reach a thermodynamic minimum. The final zeta potential value given is an average of the number of experiments conducted at 298 K, experiments were conducted in H2O / 5 % EtOH at a range of concentrations, exProton NMR spectroscopy titration studies:First 1.5 mL of a 0.01 M solution of receptor (SSA) was prepared. Of this solution, 0.5 mL was added to an NMR tube, which was then sealed with an airtight suba seal. The remaining 1.0 mL of the receptor solution was used to make a 0.15 M solution of the TBA salt of the anion. The anion / receptor solution was titrated into the NMR tube in small aliquots and a1H NMR spectrum taken after each addition. This allows the concentration of the anion in the NMR tube to increase whilst the concentration of receptor to remain constant. Chemical shifts for each1H NMR spectrum were recorded in ppm and calibrated to the solvent peak set. Bindfit v0.5 is then used to interpret the data to solve the binding constant(s). Click or tap here to enter text. Where SSAs are supplied as coformulations, they are present in a 1:1 molecular ratio, with the total molecular concentration equal to that of the of the experiment containing a single agent only.Formation of the heterogenous SSA co-formulation: Homogenous solutions of each SSA (2.78 mM) were combined at equal volumes. The heterogeneous solution then underwent an annealing process in which the solution was heated to 50 °C for 60 seconds before being allowed to cool to room temperature.Preparation of vesicles: All lipids were purchased as powder stocks from Avanti Polar Lipids. Lipid vesicles were prepared according to standard procedures. Each phospholipid (~ 20 mg) was dissolved in chloroform: methanol 3:1 (~ 10 mL) and the solvent was removed under reduced pressure. The thin film of dried lipid was further dried in vacuo overnight. Lipid films were resuspended in the desired buffer and subjected to 10 freeze-thaw cycles in liquid nitrogen and extruded 19 times through a 200 nm polycarbonate membrane.Preparation of calcein loaded vesicles: Lipid films were resuspended in calcein dye (70 mg calcein,Case Ref. P319WO IPTector®865 μL deionized H₂O, 135 μL NaOH (2 M), with NaOH added dropwise until calcein dissolved) and subjected to 10 freeze-thaw cycles in liquid nitrogen and extruded 19 times through a 200 nm polycarbonate membrane. The suspension was run down a size exclusion column packed with sephadex G-50 using 1 X PBS. The hydrodynamic dimeter of the lipids was monitored using an Anton Paar Litesizer™ 500 to ensure the separation of free and entrapped calcein. Phospholipid concentration was adjusted to 60 pM for vesicle leakage experiments.Vesicle leakage assay: Black bottom 96-well plates were prepared by serially diluting desired SSA in a EtOH: H₂O 1:19 solution across the plate, the appropriate lipid solution (100 μL, 60 μM) was added to each well to give a total well volume of 200 pL. A solution of 2 % Triton X-100 was used for a 100 % lysis control and a EtOH: H₂O 1:19 solution was used as a 0 % lysis control. Fluorescent measurements were taken at 25 °C using an excitation value of 495 nm and an optimised gain of 800. Data were acquired in endpoint mode. All experiments were repeated in triplicate to ensure experimental reproducibility.Preparation of DPH fluorescent labelled vesicles: Lipid films were resuspended in buffer (150 mM KCI, 10 mM HEPES, pH 7.4, 2 mM EGTA) and subjected to 10 freeze-thaw cycles in liquid nitrogen and extruded 20 times through a 200 nm polycarbonate membrane. For fluorescent labelling, the desired vesicles were pre-incubated with 55 (10 pM) at 60 °C for 1 hour.Membrane fluidity assay: Black bottom 96-well plates were prepared by serially diluting desired SSA in a EtOH: H₂O 1:19 solution across the plate, the appropriate DPH labelled vesicles (100 μL, 6 0 μM) were added to each well to give a total well volume of 200 pL. FP measurements were taken at 25 °C using a 355 nm filter for excitation and a 430 m for emission. The DPH labelled vesicles were set to a FP value of 100 mP. Data were acquired in endpoint mode. All experiments were repeated in triplicate to ensure experimental reproducibility.Fluorescence polarisation: Lipid films were resuspended in Buffer (150 mM KCI, 10 mM HEPES, pH 7.4, 2 mM EGTA) and subjected to 10 freeze-thaw cycles in liquid nitrogen and extruded 21 times through a 200 nm polycarbonate membrane. Black bottom 96-well plates were prepared by serially diluting desired vesicle solution (2 mM) across the plate, the appropriate SSA solution (100 pL, 0.3 mM) was added to each well to give a total well volume of 200 pL. Fluorescent SSA solutions were set to a FP value of 100 mP. Data were acquired in endpoint mode. All experiments were repeated in triplicate to ensure experimental reproducibility.Case Ref. P319WO IPTector®Membrane permeability and adhesion assay, determining membrane adhesion factors (MAF) and permeation factors (PF): Lipid vesicles of DOPE and PG, in a 1:1 ratio by weight were prepared as described in 'preparation of vesicles' with one alteration. Here vesicles were extruded 21 times through a 1000 nm polycarbonate membrane. NMR samples were prepared containing: SSA (200 pM), 95% HEPES buffer (HEPES (10 mM), NaCI (10 mM)) and 5% D2O, and sodium trimethylsilylpropanesulfonate - DSS (1 pM). In order to calculate the PF and MAF the paramagnetic relaxation enhancement agent (PRE), gadodiamide (1 mM) was added to the NMR mixture. Where lipid vesicles were present in the NMR mixture, vesicles were added at a concentration of 0.1 mg / mL. All1H CPMG NMR spectra were collected with a spin-lock time of 150 mS on a Bruker Avance III spectrometer equipped with a cold probe at a proton frequency of 600 MHz and recorded at 298 K. Peak intensities were measured using MestReNova (Mestrelab Research). The MAF and PF were determined using previously published method (Chem. Commun., 2024, 60, 11160-11163). For individual SSAs, MAF and PF were calculated using all aromatic CH resonances. For SSA coformulations an average MAF and PF were calculated using all aromatic CH resonances. For individual components of the SSA co-formulations, the resonances for the different SSA anions were treated independently of one another. Experiments were performed on a Bruker 600 AVIII NMR machine, fitted with a cryoprobe.Antibiofilm experimental: The reference strains, Pseudomonas aeruginosa PAO1, Candida albicans SC5314, Candida auris MRU1930, MRU1965 and MRU2529 were used in this study. The strains were stored at -80 °C in nutrient broth (NB) (1 gL-1meat extract, 2 gL-1yeast extract, 5 gL-1peptone, and 8 gL-1sodium chloride) supplemented with 25 and 15 % (v / v) glycerol, respectively. Before each assay, the P. aeruginosa, C. albicans and C. auris strains were cultured from the frozen stock onto nutrient agar (NA) (1 gL-1malt extract, 2 gL-1yeast extract, 5 gL-1peptone, 8 gL-1sodium chloride, 20 gL-1agar) and yeast malt extract (YM) agar (3 gL-1malt extract, 3 gL-1yeast extract, 5 gL-1peptone, 10 gL-1glucose). For every experiment, a fresh (preinoculum) culture of P. aeruginosa was prepared by transferring a single colony from the maintained plates into 10 mL of NB and incubating at 37 °C with shaking (150 rpm (RPM)) for 24 hours. Similarly, a fresh (preinoculum) culture of C. albicans / C. auris was prepared into 5 mL of yeast nitrogen base (YNB) broth (6.7 gL-1YNB, 10 gL-1glucose) and incubated at 30 °C for 24 hours. For all assays, filter-sterilized (0.22 pm nitrocellulose filter, ABLUO, GVS, USA) RPMI 1640 medium with l-glutamine and sodium bicarbonate (Sigma-Aldrich, UK) at pH 7.0, was used.Metabolic assay (antibiofilm activity): The preinoculum was washed three times with phosphate-Case Ref. P319WO IPTector®buffered saline (PBS) (0.2 gL-1potassium chloride, 0.2 gL-1potassium dihydrogen phosphate, 1.15 gL“1disodium hydrogen phosphate, 8 gL-1sodium chloride) (Oxoid, UK) at pH 7.3 and standardized to an optical density (OD) OD595 of 0.5 for P. aeruginosa, 1 x 106cells mL-1for C. albicans, and for C. auris.The standardized cell suspension was dispensed into a 96-well flat-bottom culture (microtiter) plate (Greiner Bio-One, Germany) (250 pL total volume per well) together with a twofold dilution series of each SSA to achieve a final concentration range. The microtiter plates were incubated for 48 hours at 37 °C and OD measurement (at a wavelength of 595 nm, OD595) (EZ Read 800 Research, Biochrom, England) was performed. To cultivate polymicrobial biofilms, the methods described for the monomicrobial biofilm models were used. However, when standardizing the cell solutions needed for the inoculation of a microtiter plate, P. aeruginosa and C. albicans / C. auris cell suspensions were adjusted to an OD595 of 0.1 and 2 x 106cells mL-1, respectively. After incubation, an indirect and semiquantitative measure of biofilm formation was performed, using a 2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide (XTT) colorimetric reduction assay as described using 1 gL-1of filter-sterilized (0.22 pm nitrocellulose filter) XTT salt (Sigma-Aldrich, UK), dissolved in PBS, and supplemented with 1 mM menadione in acetone. The supernatant from the 96-well plates was discarded, the wells washed twice with 200 pL of PBS before 50 pL of XTT-menadione solution was introduced to each well. The plates were incubated for 3 hours at 37 °C in the dark and the absorbance measured at 492 nm using a microtiter plate reader (EZ Read 800 Research, Biochrom, England). Furthermore, when considering the antimicrobial potential of quaternary ammonium compounds, such as TBA, the counterion of the selected SSAs, the antibiofilm activity of TBA chloride was also evaluated. This was repeated five times. Additionally, appropriate controls were included for known antimicrobials, colistin, and fluconazole, respectively.Confocal Laser Scanning Microscopy (CLSM): C. albicans was cultivated on a glass coverslip in a six-well microtiter plate (Greiner Bio-One, Germany) for 2 hours at 37 °C. After incubation the microscope slide was placed in the confocal microscope (ZEISS LSM 900 with an AiryScan 2 confocal laser scanning microscope (ZEISS, Germany)) and SSA added at an excitation / emission wavelength of 365 / 461 nm (using the DAPI filter setting). Visualisation occurred for 10 minutes.Statistical Analyses for biofilm experiments: For all quantitative experiments, the averages and standard deviations were calculated. Data produced were analysed using a Student t test to establish statistically significant differences between data sets. A p-value of <0.05 was considered significant.Case Ref. P319WO IPTector®Molecular modelling and simulation:

[0278] Structures of the compounds were drawn in ChemDraw (version 22.0.0) and were imported into Chem3D (version 22.2.0). MM2 energy minimisations were run, giving the dipole-dipole energy and total energy in kcal / mol. The length of the compound was obtained by measuring the distance between the two atoms furthest away from each other, given in A and converted to nm.Low level in silico modelling:

[0279] Computational calculations to identify primary hydrogen bond donating and accepting sites were conducted in line with studies reported by Hunter using Spartan 20 v1.1.4 (C. A. Hunter, Angew. Chem. Int. Ed., 2004, 43, 5310-5324.). Calculations were performed using semi-empirical PM6 methods, after energy minimisation calculations, to identify Emaxand Eminvalues. PM6 was used over AMI in line with research conducted by Stewart (J. J. P. Stewart, J. Mol. Model, 2007, 13, 1173-1213).Patch clampSolutions and compound preparation:

[0280] All solvents and starting materials were purchased from known chemical suppliers. Compound stock solutions were prepared in DMSO. Buffer A consisted of KCI (489 mM) and NaOAc (5 mM) with a pH of 5.5 and an ionic strength of 500 mM. Buffer B consisted of Na₂SO₄ (167 mM) and NaOAc (5 mM) with a pH of 5.5 and an ionic strength of 500 mM.Vesicle formation: Giant unilamellar Vesicles (GUVS) were prepared through the electro formation method using a Nanion VesiclePrepPro. A solution of 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) (10 mM) with 10 % cholesterol in chloroform (20 pL) was deposited onto an ITO coated glass surface (surface coated glass slide) and allowed to evaporate to create a dehydrated thin lipid film. A greased O-ring was then placed around the dehydrated lipid film to create a seal. This volume was then filled with a sucrose solution (275 pL, 200 mM) and then sealed with a second ITO coated glass slide. An AC current (3 V / 5 Hz) was then applied. After 2 hrs, the voltage and frequency were reduced in steps over 30 mins to 1.6 V / 1 Hz, and held for 1 hr before being reduced to 0 V / 0 Hz over a further 30 min period.Patch clamp measurements: Experiments were performed using a Port-a-Patch system (Nanion Technologies) with borosilicate glass chips that contain an aperture diameter of approximately 1 pm. Buffer A (5 pL) was placed on the underside of the chip over the aperture, The chip was then placed into the port-a-patch system. Buffer A (15 pL) was then added to the top of the chip and 5 pL of the GUV stock solution added. The GUVS were the positioned over the aperture in the chip by application of negative pressure (-30 mbar). When a GUV is positioned over the aperture in the chip, it bursts, leaving a planar bilayer as a seal. The negative pressure is then removed, leaving the planar bilayer inCase Ref. P319WO IPTector®place over the aperture. The external buffer A was the exchanged with buffer B, through four simultaneous buffer exchanges (4 x 15 pL). The holding potential was set to ±50 or ±100 mV depending on the experiment. The compound in a DMSO:buffer B 1:50 mixture (5 pL) was then added at t = 30 s, to give a final experimental concentration of 0.1 - 1 mM as appropriate. The number of sweeps was set to 1000, the sweep interval to 0.00 s, sample interval to 1.00 ms and the protocol was run for a total of 5 mins. Each experiment was repeated a minimum of three time to ensure reproducibility.Data analysis: Initially the.asc files generated from the patch clamp experiment were curated into the format - internal_molecule_id / experimental_conditions / experimentjd / files_ending_experiment_id.asc.

[0281] Files were read and converted from.asc files to csv files using Python scripts. The process combined all.asc files associated with an experiment into a single csv which can be read in and manipulated using Python. The experiments had been recorded at different resolutions; therefore time rather than number of points has been used in the analysis. Two methods were used to categorise the time series patch clamp data into discrete states for analysis, method 1(to get an understanding of the true baseline and small current changes) and method 2 (to get a macroscopic view of each experiment).Method 1 involved finding the mean of the first 200 points for each experiment. The system was defined as being in an event state if a percentage of datapoints are above the baseline threshold (mean + standard deviation for each experiment). To do this, a convolution operation was applied to a masked version of the patch clamp current data with a vector containing only ones set to length 20. The masked version of the dataset set each datapoint 1 if the datapoint was above the baseline threshold and 0 if it was below. The convolution returned a look forward window with the number of points above and below the baseline which was converted to a percentage and checked against a threshold (set at 70 %). This set to 1 (event) if above and 0 (baseline) if below. Method 1 was used as a tool in analysis to pick and plot all events. These events were then post-processed and those events with a mean current below the standard deviation of the baseline or those that are too short-lived (less than 0.2 seconds by default) were removed.Method 2 was a data binning approach used to classify the data into 4 event types (though these have been scaled for the variable current experiments) - 0: baseline (-inf A to 0.5e-10 A), 1: events (0.5e-10 A to 5e-10 A), 3: events where the current readings surpass the detector's measurable range (4.9e-8 A to inf A), and 2: events where current is between 5e-10 A and 4.9e-8 A. These current bins are as stated above for + 100 mV, halved for + 50 mV experiments, and made negative for -100 mV and for -50 mV. When the current is around the cutoffs, the noise causes the data to be categorised as above and below and threshold, these times are set to the minimum resolution of the experiment.Case Ref. P319WO IPTector®The data was processed using a PatchClampData class coded in Python which gives the ability to run these thresholding methods and generate the plots used within this paper and ESI. It has also allowed a summary table of the experiments including the total time of each experiment, the time in each event state and the bins used to threshold each experiment to be generated. The code for this project is available on GitHub - https: / / github.com / ta1u18 / ssa_patch_clamp_data_analysis and original data, processed data and figures available as part of the supplementary information.Ion transport studiesPreparation of vesicles: A stock solution of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) was prepared by dissolving 1 g of POPC in 35 mL de-acidified CHCI3. The CHCl₃ was passed through a column of basic alumina before use. This solution was stored at -20 °C when not in use.A lipid film of POPC was prepared by adding 1-4 mL of the above stock solution into a pre-weighed falcon tube and gently removing the CHCl₃ under a stream of nitrogen. The lipid film was dried under vacuum overnight. The lipid film was re-hydrated by vortexing with the desired internal buffer solution, then subjected to 9 freeze-thaw cycles. Typically the lipids were not thawed on the last cycle and were stored at -20 °C for use at a later date.The lipid was thawed to room temperature and allowed to stand for 30 mins, then extruded 27 times through a 200 nm polycarbonate membrane. The resulting unilamellar vesicles were dialysed against the desired external buffer solution for a minimum of 2 hours.Transport experiments protocol: The vesicles prepared as described above were diluted to a known volume of 5 mL using the desired external buffer solution. This solution was further diluted into individual 5 mL samples for testing at a final lipid concentration of 1 mM. The transporter was added to the vesicles from a stock solution in DMSO (5 mM) after 30 s in order to initiate the transport and any resulting chloride efflux was monitored using a chloride selective electrode (ISE - Cole Parmer). At the end of the experiment, the vesicles were lysed by the addition of 40 pLTriton-X 100 (10 wt% in H2O). The final electrode reading was taken 2 mins later and used to calibrate 100% chloride efflux. Each experiment was performed 3 times.MIC assayPreparation of bacterial plates: Sterile TSB agar plates were streaked using the desired bacteria [Staphylococcus aureus (9144, 13616, USA 300 and 1199B), Enterococcus faecalis (NCTC 775 and 12201) and Enterococcus faecium (NCTC 12204)] and incubated at 37 °C overnight.Preparation of inoculum: An initial culture was made up by inoculating TSB (5 mL) with 3 single colonies of the desired bacteria under sterile conditions and incubated at 37 °C overnight. The optical density at 600 nm (OD₆₀₀) was adjusted using sterile distilled H₂O (dH₂O) to equal ~1 x 106bacteria / mL.Preparation of 96-well microplate: A solution of each compound in H2O / 5 % EtOH was made at a topCase Ref. P319WO IPTector®concentration of 5.56 mM on the day of experiment. The compound solution (200 pL) was added to and diluted 50:50 down the plate. The bacterial suspension (100 pL) was dispensed into each well under sterile conditions. The plates were sealed using parafilm and incubated at 37 °C for 20 hours, after which the OD₆₀₀ was measured.Haemolysis assayPreparation of 96-well plate: Protocol modified from Travis et al.4Heparinised human red blood cells collected from a volunteer and washed three times in PBS (pH 7.4). Compounds were serial diluted using PBS buffer across a 96-well V bottom Greiner plate. Negative control (100 pL PBS), and positive control (100 pL 0.1 % (v / v) Triton-X-100) were added to the plate. 100 pL of 10 % (v / v) of blood suspension added to all wells. Plates were incubated for 60 minutes at 37 °C. After incubation, plates were centrifuged for 15 minutes at 4680 rpm. The resultant supernatant was then transferred to a 96-well flat bottom plate and absorbance read at 540 nm (Fluostar Omega). Percent haemolysis was calculated using the formula as shown in Equation SI.Equation SI - Percentage haemolysis calculation.Haemolysis (%) = (Asample− Anegative) / (ATriton− Anegative) × 100Triton ^negativeToxicity assayG. mellonella treatment assay: G. mellonella larvae were purchased from Livefood UK Ltd. (Rooks Bridge, UK) and maintained on wood chips in the dark at 15 °C until use. Galleria larvae were injected with 10 pL of compound in H2O / 5 % EtOH, incubated at 37 °C for 5 days and the deaths counted. Groups of 10 Galleria were injected per compound.GI50determination assaysCell culture: The RPE-1 non-cancerous hTERT human retinal pigment epithelial, A2780 human ovarian carcinoma, and A2780 CisR cisplatin resistant human ovarian carcinoma cell lines were donated with thanks from the Institute of Cancer Research. The A2780 and A2780 CisR cell lines were cultured in IMDM supplemented with 10 % FBS at 37 °C and 5 % CO2, while the RPE-1 cell line was cultured in a 1:1 mixture of Dulbecco's Modified Eagle Medium and Ham's F-12 Nutrient Mixture (DMEM / F-12) at 37 °C and 5 % CO2.SRB assay: Cells were seeded into 96-well plates at 800 cells per well / cpw (A2780), 1600 cpw (A2780 CisR) or 400 cpw (RPE-1) in cell culture medium and cultured for 48 hours followed by addition of each compound over an eight-point concentration range (each concentration in triplicate). Plates were then cultured for a further 96 hours, after which the cell culture medium was removed from each well and the cells fixed with addition of 70 pL / well of 10 % (w / v) trichloroacetic acid (TCA) in distilled water followed by incubation for 30 minutes at RT. Each plate was then washed with distilled water fiveCase Ref. P319WO IPTector®times before addition of 70 μL SRB dye (0.4 % (w / v) SRB dye (ThermoFisher Scientific, USA) solubilised in 1 % (v / v) acetic acid / dH₂O) and incubated for 30 minutes at RT, followed by washing three times with 1 % (v / v) acetic acid and drying in a 37 °C oven overnight. Once dry, 100 μl of 10 mM Tris-base (ThermoFisher Scientific, USA) was added to each well and plates put on a microplate shaker for 10 minutes at 200 rpm to solubilize the dye. Absorbance values were then read at wavelength 490 nm on a Victor X4 multi-label plate reader (PerkinElmer Life Sciences, USA), data analysed using Microsoft Excel and graphs produced using GraphPad Prism 9.0 and the GI50value calculated.In vitro Drug Metabolism and Pharmacokinetics studiesGeneral note: The percentage of a compound recovered during the analysis process should be 100 %; any deviation from this value indicates experimental limitations such as unintended potential nonspecific binding events to experimental equipment, solubility issues, etc.Protein Binding Measurements in Human Plasma by Using Equilibrium Dialysis Method:1 Preparation of compound working solutionsThe working solution of test and control compound were prepared in DMSO at the concentration of 1 mM.2 Preparation of buffer solution pH 7.4A basic solution was prepared by dissolving 14.2 g / L Na₂HPO₄ and 8.77 g / L NaCl in deionized water and the solution could be stored at 4 °C for up to 7 days. An acidic solution was prepared by dissolving 12.0 g / L NaH₂PO₄ and 8.77 g / L NaCl in dH₂O and the solution could be stored at 4 °C for up to 7 days. The basic solution was titrated with the acidic solution to pH 7.4 and store at 4°C for up to 7 days. pH was checked on the day of experiment and was adjusted if outside specification of 7.4 ± 0.1.3 Preparation of plasmaSet the temperature of water bath to 37 °C. Thaw the frozen Plasma (stored at -80°C) immediately in a 37 °C water bath.4 Preparation of operation plateSoak the dialysis membranes in ultrapure water for 60 minutes to separate strips, then in 20 % ethanol for 20 minutes, finally in dialysis buffer for 20 minutes. Load the prepared membranes into the dialysis device and install the device again following manufacturers guidelines. Turn on air bath and allow to pre-heat to 37 °C.5 Preparation of control sample at 0 hourAdd 597 pL of blank plasma solution into each vial of a new plastic plate or separate plastic tube by addition of 3 pL of the working solution of test compound, vortex at 1000 rpm for 2 minutes. The final percent volume of organic solvent is 0.5 % and the final concentration for test compound is 5Case Ref. P319WO IPTector®pM. Immediately transfer 50 pL of the spiked plasma solution suspension to a 96-well plate to act as T=0 control sample. The samples are treated the same as the samples after incubation. Place all remaining spiked plasma solution in the incubator for the duration of the study.6 Stability determination of test in plasma solutionAt the same time, the remaining spiked plasma solution sample in the plastic plate or separate plastic tube is incubated for 6 hours at 37 °C with 5 % CO2 in the CO2 incubator.At T=6 hours, transfer 50 pL of the original spiked plasma solution suspension to the 96-well plate for analysis.7 Procedure for equilibrium dialysisAssemble the dialysis set up following the manufacturer's instructions. Load cells with 120 pL of plasma sample and dialyzed against equal volume of dialysis buffer (PBS). The assay is performed in duplicate. Cover the unit with gas permeable lid and incubate for 6 hours at 37 °C at 100 rpm with 5 % CO2 on an orbital shaker in the CO2 incubator. At the end of incubation, remove lid and pipette 50 pL of post-dialysis samples from both buffer and plasma solution chambers into separated 96- well plate for analysis, respectively.8 Procedure for sample preparationAdd 50 pL of plasma solution to the buffer samples, and an equal volume of PBS to the collected plasma solution samples. Shake the plate at 1000 rpm for 2 minutes and add 400 pL of acetonitrile containing an appropriate internal standard (IS) to precipitate protein and release compound. Vortex at 1000 rpm for 10 minutes. Centrifuge for 30 minutes at 3,220 g. Then transfer 100 pL of the supernatant to new 96-well plates for analysis. Add 100 pL of distilled water to each sample and mix for analysis by LC-MS / MS.9 Data AnalysisAll calculations are carried out using Microsoft Excel. Determine the concentrations of test compound and control compound in the buffer and plasma solution chambers. Calculate the percentages of test compound(s) and control compound bound as follows:% Unbound = (Area ratio buffer chamber / Area ratio plasma solution chamber) × 100% Bound = 100 - % Unbound% Recovery = (Area ratio buffer chamber + Area ratio plasma solution chamber) / (Area ratio Total sample) × 100 % Remaining = Area ratio 6hr / Area ratio 0hr × 100Kinetic Solubility Determination in PBS pH 7.4:Study Procedure1. Preparation of stock solutionsCase Ref. P319WO IPTector®The stock solutions of test compound and control compounds (progesterone and diclofenac) were prepared in DMSO at the concentrations of 10 mM.2. Procedure for solubility determination15 pL of stock solution (10 mM) of each sample was placed in order into their proper 96-well rack.485 pL of buffer was added into each vial of the cap-less Solubility Sample plate. The assay was performed in duplicate. Add one stir stick to each vial and seal using a moulded PTFE / Silicone plug. Then the solubility sample plate was transferred to the Eppendorf Thermomixer Comfort plate shaker and shaken at 25 °C at 1100 rpm for 2 hours. After completion of the 2 hours, plugs were removed and the stir sticks were removed using a big magnet, the samples from the Solubility Sample plate were transferred into the filter plate. Using the Vacuum Manifold, all the samples were filtered. Aliquot of 5 pL and 5 pL DMSO were taken from the filtrate followed by addition of 490 pL of a mixture of H2O and acetonitrile containing internal standard (1:1) or methyl alcohol. A certain proportion of a mixture of H2O and acetonitrile containing internal standard (1:1) or methyl alcohol was used to dilute the diluent according to the peak shape. The dilution factor was changed according to the solubility values and the LC-MS signal response.3. Preparation of 300 pM standards (STD)From the 10 mM DMSO STD plate, 6 pL was transferred into the remaining empty plate, and then 194 pL of DMSO was added to that plate to have a STD concentration of 300 pM. From the 300 pM DMSO STD plate, 5 pL DMSO STD and 5 pL buffer were transferred into the remaining empty plate, and then 490 pL of a mixture of H2O and acetonitrile containing internal standard (1:1) or methyl alcohol was added to that plate to have a final STD concentration of 3 pM. A certain proportion of mixture of H2O and acetonitrile containing internal standard (1:1) or methyl alcohol was used to dilute the diluent according to the peak shape. The concentrations of the standard samples were changed according to the LC-MS signal response.4. Procedure for sample analysisThe plate was placed into the well plate autosampler. The samples were evaluated by LC-MS / MS analysis.5. Data analysisAll calculations were carried out using Microsoft Excel.The filtrate was analysed and quantified against a standard of known concentration using LC coupled with mass spectral peak identification and quantitation. Solubility values of the test compound and control compound were calculated as follows:[Sample]=Area rati0SamplexDFSamplex STDArea ratio- ill -Case Ref. P319WO IPTector®DF means the dilution factor.Any value of the compounds that was not within the specified limits was rejected and the experiment was repeated.Metabolic Stability in Rat and Mouse Liver Microsomes:Study Design1. The master solution was prepared according to Table A.Table A. Preparation of master solutionReagent Stock Volume Final Concentration ConcentrationPhosphate 100 mM 216.25 pL 100 mMbufferMicrosomes 20 mg / mL 6.25 pL 0.5 mg / mL2. Two separated experiments were performed as follows, a) With Cofactors (NADPH): 25 pL of 10 mM NADPH was added to the incubations. The final concentrations of microsomes and NADPH were 0.5 mg / mL and 1 mM, respectively, b) Without Cofactors (NADPH): 25 pL of 100 mM Phosphate buffer was added to the incubations. The final concentration of microsomes was 0.5 mg / mL. The mixture was pre-warmed at 37 °C for 10 minutes.3. The reaction was started with the addition of 2.5 pL of 100 pM control compound or test compound solutions. Verapamil was used as positive control in this study. The final concentration of test or control compound was 1 pM. The incubation solution was incubated in water batch at 37 °C.4. Aliquots of 30 pL were taken from the reaction solution at 0.5, 5, 15, 30 and 60 minutes. The reaction was stopped by the addition of 5 volumes of cold acetonitrile with IS (100 nM alprazolam, 200 nM caffeine and 100 nM tolbutamide). Samples were centrifuged at 3, 220 g for 40 minutes. Aliquot of 100 pL of the supernatant was mixed with 100 pL of ultra-pure H2O and then used for LC- MS / MS analysis.5. Data AnalysisAll calculations were carried out using Microsoft Excel.Peak areas were determined from extracted ion chromatograms. The slope value, k, was determined by linear regression of the natural logarithm of the remaining percentage of the parent drug vs. incubation time curve.The in vitro half-life (in vitro ti / 2) was determined from the slope value:Case Ref. P319WO IPTector®m vitro tij® = - (0.693 / Ik)Conversion of the in vitro ti / 2 (min) into the in vitro intrinsic clearance (in vitro CLint, in pL / min / mg protein) was done using the following equation (mean of duplicate determinations):0.693 volume of incubation t L) m wtro CL« = ( — ) • ( — )(tie) amount of proteins (rr»g)The calculations of Scale-up CLint (mL / min / kg), Predicted Hepatic CLH(mL / min / kg) and Hepatic Extraction Ratio (ER) were done using the following equation:Scale-up CLjnt = (0.693 / Ti / z) x (l / (microsomal protein concentration (0.5 mg / mL))) x Scaling Factors (Table B)Predicted Hepatic CLH= (QH x Scale-up CLint x fub) / (QH + Scale-up CLint x fub),ER = Predicted Hepatic CLH / QHwhere QH is the hepatic blood flow (mL / min / kg) (Table B), fub is the fraction of unbound drug in plasma which is assumed to be 1.Table B. Scaling factors for intrinsic clearance prediction in human, monkey, dog, rat and mouse microsomesSpecies Microsomal Liver Weight per Scaling Factor3Hepatic Blood Protein Kilogram of Body Flow per Gram of Liver Weight (mL / min / kg) Human 40 25.7 1028 21 Monkey 40 30 1200 44Dog 55 32 1760 31Rat 61 40 2440 68 Mouse 47 88 4136 90aScaling Factor = (microsomal protein per gram of liver) x (liver weight per kilogram of body weight) 6. Data Processing RulesThe rules for data processing are shown in Table C.Table C. The rules on data processingRemaining % Processing Rules>80% at 60 min If T-test with p<0.05 is obtained, report the calculated CLint value; When the calculated Clint value <7.50, then report <7.50 instead of calculated value. If T-test with p<0.05 is not obtained, then report <7.50 for CLint value when all the other data points fall in the range of 80 %~120 % (one data point within theCase Ref. P319WO IPTector®range of 70 %~130 % is accepted, otherwise the experiment should be repeated).< 80 % at 60 min Always remove from the calculation all points with < 10 % left of 0.5 min sample, but leave at least 2 pointsIf T-test with p<0.05 is obtained, report the calculated CLint value.If T-test with p<0.05 is not obtained, the experiment must be repeated.Bidirectional Permeability in Caco-2 Cell Line:Study Design1. Preparation of Caco-2 Cells1) 50 pL and 25 mL of cell culture medium were added to each well of the Transwell insert and reservoir, respectively. And then the HTS transwell plates were incubated at 37 °C, 5 % CO2 for 1 hour before cell seeding.2) Caco-2 cells were diluted to 6.86xl05cells / mL with culture medium and 50 pL of cell suspension were dispensed into the filter well of the 96-well HTS Transwell plate. Cells were cultivated for 14- 18 days in a cell culture incubator at 37 °C, 5 % CO2, 95 % relative humidity. Cell culture medium was replaced every other day, beginning no later than 24 hours after initial plating.2. Assessment of Cell Monolayer Integrity1) Medium was removed from the reservoir and each Transwell insert and replaced with prewarmed fresh culture medium.2) Transepithelial electrical resistance (TEER) across the monolayer was measured using Millicell Epithelial Volt-Ohm measuring system (Millipore, USA).3) The Plate was returned to the incubator once the measurement was done.The TEER value was calculated according to the following equation:TEER measurement (ohms) x Area of membrane (cm2) = TEER value (ohm»cm2)TEER value should be greater than 230 ohm»cm2, which indicates the well-qualified Caco-2 monolayer.3. Preparation of Solutions1) Prepare stock solutions of test compound in DMSO at 1 mM and dilute with HBSS (10 mM HEPES, pH 7.4) to get 5 pM working solution. Metoprolol and Digoxin are used as control. Prepare stock solutions of control in DMSO at 2 mM and dilute with HBSS (10 mM HEPES, pH 7.4) to get 10 pM working solution.4. Performing the SSA Transport AssayCase Ref. P319WO IPTector®1) Remove the Caco-2 plate from the incubator. Wash the monolayer twice with pre-warmed HBSS (10 mM HEPES, pH 7.4). Then incubate the plate at 37 °C for 30 minutes.2) To determine the rate of drug transport in the apical to basolateral direction. Add 125 pL of the working solution to the Transwell insert (apical compartment), and transfer 50 pL sample immediately from the apical compartment to 200 pL of quenching solvents containing IS (100 nM alprazolam, 200 nM Caffeine and 100 nM tolbutamide) in a new 96-well plate as the initial donor sample (A-B). Vortex at 1000 rpm for 10 minutes. Fill the wells in the receiver plate (basolateral compartment) with 235 pL of transport buffer.3) To determine the rate of drug transport in the basolateral to apical direction. Add 285 pL of the working solution to the receiver plate wells (basolateral compartment), and transfer 50 pL sample immediately from the basolateral compartment to 200 pL of quenching solvents containing IS (100 nM alprazolam, 200 nM Caffeine and 100 nM tolbutamide) in a new 96-well plate as the initial donor sample (B-A). Vortex at 1000 rpm for 10 minutes. Fill the Transwell insert (apical compartment) with 75 pL of transport buffer. The apical to basolateral direction and the basolateral to apical direction need to be done at the same time.4) The plates were incubated at 37 °C for 2 hours.5) At the end of the incubation, 50 pL samples from donor sides (apical compartment for Ap-> BI flux, and basolateral compartment for Bl— > Ap) and receiver sides (basolateral compartment for Ap-> BI flux, and apical compartment for Bl— > Ap) were transferred to wells of a new 96-well plate, followed by the addition of 4 volume of quenching solvents containing IS (100 nM alprazolam, 200 nM Caffeine and 100 nM tolbutamide). All samples were Vortexed for 10 minutes, and then centrifuge the quenching plates of each time point for 30 minutes at 4,000 rpm and 4°C. Then transfer the supernatant was mixed with an appropriate volume of ultra-pure water before LC-MS / MS analysis.6) To determine the Lucifer Yellow leakage after 2-hour transport period, stock solution of Lucifer yellow was prepared in DMSO and diluted with HBSS (10 mM HEPES, pH 7.4) to reach the final concentration of 100 pM. 100 pL of the Lucifer yellow solution was added to each Transwell insert (apical compartment), followed by filling the wells in the receiver plate (basolateral compartment) with 300 pL of HBSS (10 mM HEPES, pH 7.4). The plates were Incubated at 37 °C for 30 mins. 80 pL samples were removed directly from the apical and basolateral wells (using the basolateral access holes) and transferred to wells of new 96 wells plates. The Lucifer Yellow fluorescence (to monitor monolayer integrity) signal was measured in a fluorescence plate reader at 480 nM excitation and 530 nM emission.5. Data analysisCase Ref. P319WO IPTector®All calculations are carried out using Microsoft Excel. Peak areas are determined from extracted ion chromatograms.Lucifer yellow leakage of monolayer can be calculated using the following equation:LY Leakage = - -Iacaptor X°'3- -1 x 100 % / acceptor X 0.3 + I totor X 0. 1 JWhere Iacceptor is the fluorescence intensity in the acceptor well (0.3 mL), and Idonor is the fluorescence intensity in the donor well (0.1 mL) and expressed as % leakage. Lucifer yellow percentage amount transported values should be less than 1.5 %. However, if the lucifer yellow percentage amount transported value for a particular transwell is higher than 1.5 but the determined digoxin Papp in that transwell is qualitatively similar to that determined in the replicate transwells then, based upon the scientific judgement of the responsible scientist, the monolayer is considered acceptable.Apparent permeability (Papp) can be calculated for drug transport assays using the following equation:D_ ^Av[^™^Lccepto(r — - "r - 1 -appArea x time drug,, Where: Pappis apparent permeability (cm / s x 10'6)V is the volume (in mL) in the acceptor wellArea is the surface area of the membrane (0.143 cm2for Transwell-96 Well Permeable Supports) time is the total transport time in seconds.Efflux ratio can be determined using the following equation:pEfflux Ratio (B-A)Where Papp< B-A) indicates the apparent permeability coefficient in basolateral to apical direction, and PaPP(A-B) indicates the apparent permeability coefficient in apical to basolateral direction.Example 2 - Physicochemical characteristics of SSA self-association eventsPhysicochemical properties

[0282] The self-associative properties of selected SSAs were first explored in DMSO-dg (Table 1), as within polar organic solvents, SSAs have previously been shown to produce hydrogen bonded anionic dimers. This enables the quantification of parameters such as SSA anion dimerization constant to be established, which we have previously shown to correlate with critical aggregation concentration (CAC) values obtained from aqueous solutions of that same SSA. Here, an increase in SSA anionic dimerization constant was shown to correlate with a decreasing CAC, when weighted by a calculatedCase Ref. P319WO IPTector®LogP value, obtained for the SSA anion hydrophilic moiety.

[0283] Initially, quantitative (Q)1H NMR spectroscopy experiments were undertaken in DMSO-d61% DCM, in which the DCM was used as an internal standard to identify the presence of either low or high order SSA self-associated aggregates. Here, low-order SSA self-associated aggregates are defined as those directly observable using traditional solution state1H NMR spectroscopy (such as the formation of hydrogen bonded anionic dimers), whereas high-order SSA self-associated aggregates are defined as the exact opposite. To determine the presence of low or high-order SSA aggregates, signals corresponding to the anionic or cationic component of the SSA are integrated against those of the internal DCM standard. An apparent 'loss' of signal indicates the presence of larger, NMR silent aggregates, which exhibit solid-like properties and are therefore no longer visible using this experimental technique. As summarised in Table 1, at concentrations < 112 mM, SSAs 72, 73, 56 and 57 form low order self-associated aggregates in polar organic DMSO-dg solutions. However, 1:1 enantiomeric mixture of SSA72 + SSA73 and SSA56 + SSA57 also confirm the presence of higher order aggregates, providing evidence to support the hypothesis that the interaction of SSA anion enantiomers influences molecular self-association events in DMSO-dg 1 % DCM. We hypothesise that this is driven by the preferential heterogeneous hydrogen bonded complex formation between the SSA anionic components, such as those shown in Figure 2b. Here, the difference in amino acid side chain was also found to influence the relative proportion of the SSA to become incorporated into these higher order SSA aggregates. As the Kdimcalculated for homogenous solutions of SSA72, SSA73, SSA56 and SSA57 was found to be lower for SSA56 and SSA57 when compared to SSA72 and SSA73, we can discount the strength of hydrogen bonding influencing the increase in the proportion of SSA to become incorporated into these higher order structures at this concentration. Instead, we hypothesise that this increase in SSA to become incorporated into higher order aggregates to be due to a combination favourable pi-pi stacking interactions and decreased steric hinderance, afforded by the change in amino acid R-group.

[0284] Table 1: Physicochemical data produced to characterise SSA self-association events in a DMSO-dg 1 % DCM (QXH NMR spectroscopy) or DMSO-d60.5 % H2O (1H NMR DOSY or dilution study) solution. Where the SSAs are supplied as an enantiomeric mixture, they are present in a 1:1 ratio. Concentrations reported represent total molecular concentration of SSA present. All Q. 1H NMR spectroscopy experiments were conducted with a delay time (dl) of 60 s at 298 K and a concentration of 112 mM, and DCM was used as the internal standard. The values given in % represent the observed proportion of compound to becomeXH NMR spectroscopy silent. The hydrodynamic diameter of the SSA species present in DMSO-dg 0.5 % H2O were obtained throughXH NMR DOSY at a total molecular concentration of 112 mM (298 K). Kdim values are calculated viaXH NMR spectroscopy dilution study,Case Ref. P319WO IPTector®followed by subsequent fitting of these data to the equal K / dimerization self-associative binding isotherm model using Bindfit vO.5.42 Electrostatic surface potential energy maximum (Emax) and minimum (Emax) values were calculated using Spartan '20, while consensus LogP values were calculated using the Swiss ADME platform.Q NMR (%)SSA Krf / m £min £max Consensus SSA SSA C / H (nm) (M1) (kJ.mol’1) (kJ.mol’1) LogP anion cation30 0 0 1.3 41 (±1.3 %) -730 -45 2.9072 0 0 1.7 121 (±1.9 %) -728 -50 4.0873 0 0 1.7 90 (±3.2 %) -728 -49 4.0756 0 0 1.7 28 (±2.1 %) -714 -47 4.0357 0 0 1.7 23 (±2.5 %) -715 -46 4.22 72+73 68 69 1.4° b N / A N / A N / A 56+57 82 78 0.9° b N / A N / A N / Aa - Evidence for the presence of higher order aggregates also obtained, b - Not calculated due to sample heterogeneity.

[0285] Following initial Q.1H NMR spectroscopy experiments, the size of any self-associated aggregates visible using conventional solution state NMR spectroscopy techniques were determined through1H NMR DOSY (Table 1). Here the dHof the SSA anion, was calculated between 0.9-1.7 nm (Table 1). This suggests the presence of molecular dimers, such as those previously observed for analogous SSAs including SSA30. Click or tap here to enter text.

[0286] As homogenous DMSO-dg solutions of SSA72, SSA73, SSA56 and SSA57 only confirmed the presence of lower order SSA anion self-associated species, such as hydrogenous anionic dimers, in line with previous observations; Click or tap here to enter text.1H NMR spectroscopy dilution studies were performed in a DMSO-d60.5 % H2O solution to verify the presence of, and establish the magnitude of, any homogenous SSA anion hydrogen bonded self-association events. Here, the disproportional downfield change in chemical shift of the urea NH's upon increasing SSA concentration indicates intermolecular SSA anion hydrogen bonded complex formation. The fitting of these data to self-associative binding isotherms, achieved using open access, web based resources such as Bindfit, Click or tap here to enter text, result in the elucidation of appropriate self-association constants. Here our data was found to fit well to the dimerization (equal K) binding isotherm, with errors reported < ±4 % -further supporting the results of theTH NMR DOSY experiments (Table 1). The dimerization constants (Kd / m) calculated forSSA56 and SSA57 were found to be comparative in magnitude to SSA30, however, lower than those calculated for SSA72 and SSA73 (Table 1).

[0287] The reason for this change in Kdimis hypothesised to be due to the effect R-group substitution has on hydrogen bond donor / acceptor spatial positioning and the comparative basicity and acidity of the carboxylate and urea functionalities respectively. It has been shown that electrostatic surfaceCase Ref. P319WO IPTector®potential values, calculated by low level computational modelling programs such as Spartan, Click or tap here to enter text, using semi-empirical PM6 molecular modelling methods, Click or tap here to enter text, can be used to estimate the hydrogen donor / acceptor activity - and thus estimate hydrogen bonding strength. For the SSA anionic component of SSA30, SSA72, SSA73, SSA56 and SSA57, Emax and Emin values obtained from electrostatic potential maps were found to correlate with the urea NH and carboxylate residues respectively. Comparison of these data reveal Emax values to remain comparable across the series of SSAs tested. When comparing the Emin values, SSA30, SSA72 and SSA73 were found to exhibit values similar to each other and lower in magnitude than that of SSA56 and SSA57. This difference in Emin indicates the basicity of these carboxylate groups to be in enhanced in the former compared to the latter, which corresponds to the increased strength of Kdimobserved for SSA72 and SSA73 over SSA56 and SSA57. However, this does not explain the difference in Kd / mobserved for SSA30 and SSA72 or SSA73. We believe that this difference is due to the increased steric bulk of SSA72 and SSA73 in comparison to SSA30, which may preorganise the SSA anion of SSA72 and SSA73 towards dimer formation, resulting in increased Kdimvalues.

[0288] Moving from a polar organic DMSO solution to an aqueous environment, the presence of lower and higher order self-associated SSA aggregates was confirmed using quantitative1H NMR spectroscopy in a D2O / 5% EtOH solution, at a total SSA concentration of 5.56 mM, as shown in Table 2. Here EtOH was used as the internal standard, and confirmed the presence of higher order aggregates, demonstrating solid like properties for homogenous solutions of SSA30, SSA72, SSA73, SSA56 and SSA57. The slight difference in anion: cation ratio is due to increased experimental error dure to decreased SSA concentration. As expected, the proportion of the SSA incorporated into higher order aggregates remained comparable for homogenous solutions of both enantiomers, with no real difference in the proportion of SSA to be incorporated into the higher order aggregates of SSA72, SSA73, SSA56 and SSA57. However, when supplied as a 1:1 enantiomeric mixture, although the trend in increased incorporation of SSA56 + SSA57 over SSA72 + SSA73 into the higher order aggregate structure was retained from previous observations in polar organic solvents, the proportion of SSA incorporated into these higher order structures was approximately 20 % lower for the 1:1 enantiomeric mixture of SSA56 + SSA57 over the homogenous solution of those same SSAs. This supports the hypothesis that, as observed within a DMSO-dg 1% DCM solution (Table 1), the combination of SSA enantiomers influences SSA self-association events, causing the amino acid R-group to play a greater role in influencing higher order SSA aggregation events.

[0289] Table 2: Physicochemical data produced to characterise SSA self-association events in a H2O 5% EtOH or D2O 5 % EtOH (1H NMR spectroscopy only) solution. Where the SSAs are supplied as an enantiomeric mixture, they are present in a 1:1 ratio. Aggregate stability and hydrodynamic diameterCase Ref. P319WO IPTector®were obtained via zeta potential and DLS measurements respectively, at a concentration of 5.56 mM and a temperature of 298 K, following an annealing process. The hydrodynamic diameter of the aggregates listed were obtained from intensity distribution peak maxima. CAC was derived at approximately 291 K from surface tension measurements. All Q1H NMR spectroscopy experiments were conducted with a delay time (dl) of 60 s at 298 K and a concentration of 5.56 mh / l, EtOH was used as the internal standard. The values given in % represent the observed proportion of compound to become1H NMR spectroscopy silent.Q NMR (%) Zeta potential CAC Surface duSSA SSA SSA (mV) (mM)(nm) error PDI Error tension" anion cation (mN m1) 30 68 a 220 a a a -37 11.2 39.33 72 50 44 266 19 0.038 0.0028 -55 (± 3.64) b 40.92c73 52 48 174 13 0.033 0.0042 -58 (± 1.50) b 40.92c56 51 48 167 19 0.029 0.0150 -58 (± 1.64) b 39.83c57 58 53 181 53 0.026 0.0120 -66 (± 0.78) b 39.64c72+73 31 33 238 3 0.047 0.0034 -57 (± 0.65) b 38.67c56+57 56 51 146 14 0.019 0.0025 -65 (± 0.31) b 39.25ca - Data not available, b - Solubility prevented data acquisition, c - Data collected at limit of SSA solubility = 5.56 mM.

[0290] The self-associative properties of SSAs 6, 33, 39, 74, 75 and co-formulations a - j, were studied using methods described in Example 1. Initially, we quantified the strength of homogeneous SSA anion self-association events, to confirm the effect of squaramide - (thio)urea substitution. Quantitative1H NMR spectroscopy experiments detected the presence of lower order self-association events only in DMSO-dg 1% DCM solutions (112 mM). Complimentary1H NMR DOSY experiments conducted in DMSO-d60.5 % H2O solutions at 298 K confirmed the SSA anion and cation components to diffuse at different rates, providing supporting evidence towards the lack of any strong ion pairing interactions between the SSA anionic and cationic components under these experimental conditions. Calculating the hydrodynamic diameter (dH) of the SSA anion from these data confirmed the dH< 1.7 nm for SSAs 6, 33, 39, 74, 75, providing evidence for the presence with a lower order self-associated species, such as dimers. The formation of SSA anionic dimers has also been shown in the solid state through single crystal X-ray diffraction experiments. The anionic component of squaramide incorporated SSA 75 was also shown to form a hydrogen bonded dimer in the solid state through single crystal X-ray diffraction studies. Here the SSA anion dimer is stabilised through the formation of four intermolecular ureaanion hydrogen bonds. Proton NMR spectroscopy dilution study data, obtained from a DMSO-d₆ 0.5% H₂O solution, were then fitted to both the EK and CoEK self-association models using Bindfit v0.5. These data were found to fit the EK over the CoEK model, which alongside the additional NMRCase Ref. P319WO IPTector®spectroscopy data caused us to hypothesise the presence of SSA anion hydrogen bonded dimers present in a DMSO-d₆ 0.5% H₂O solution. Fitting these data to the EK binding isotherm enable the calculation of a dimerization constant (Kdim, Table 3) for SSAs 6, 33, 39 and 75. The dimerization constants calculated confirm that substitution of the sulfonate ion (6 and 39) for the carboxylate ion (33) increase Kdimby approximately 50-fold. However, substituting the thiourea (33) for the squaramide (75) functionality increases the Kdimby a further 70-fold. Therefore, confirming our initial hypothesis, that incorporating the squaramide residue within the SSA anion structure would increase the strength of SSA anion self-association events.

[0291] Table 3: Physicochemical data produced to characterise SSA self-association events in a H₂O:EtOH 19:1, D₂O:EtOH 19:1 (quantitative1H NMR spectroscopy only) or DMSO-dg / 0.5 % H2O (Kdimand1H NMR DOSY only) solution. Kdimvalues were obtained from fitting1H NMR spectroscopy dilution study data (change in chemical shift of the NH groups) to the appropriate binding isotherm model using Bindfit v0.5. Aggregate stability and dHwere obtained via zeta potential and DLS measurements respectively, at a concentration of 2.78 mM and a temperature of 298 K, following an annealing process unless otherwise stated. The dHof the aggregates listed were obtained from intensity distribution peak maxima. CAC was derived at approximately 291 K from surface tension measurements. All quantitative1H NMR spectroscopy experiments were conducted with a delay time (d₁) of 60 s at 298 K and a concentration of 2.78 mM. Co-formulations a - j quantitative1H NMR spectroscopy experiments represent combined anionic component. The values given in % represent the observed proportion of compound to become NMR silent. ST = surface tension. ZP = zeta potential Quantitative ST at2H NMR(%) CAC CAC ZPSSA du(nm) (mM) (mNm (mV) Kd / rnfM’1) Anion Cation6 36 34 184 5.6 34 -56 2.633 37 34 143 >10.0° 41b-56 10539 40 40 672 0.5 47 -86 2.774 47 42 162 >7.5° 42b-79 c75 34 38 173 >2.8° 43b-73 7636Co-formulationa 58 64 107 >10.0° 38b-42 db 68 69 187 8.1 38 -72 dc 59 65 107 >7.5° 39b-68 dd 54 67 99 >2.8° 45b-54 de 75 71 189 2.8 46 -71 df 68 70 181 >7.5° 39b-76 dg 72 70 105 >2.8° 45b-62 dh 54 66 190 >7.5° 42b-75 di 58 59 690 2.5 42 -69 dj 37 49 84 >2.8° 44b-82 da - This represents the limit of solubility preventing determination of CAC. b - Surface tension at limit of solubility. c-Slow exchange processes prevented Kdim calculation, d - Multiple component systemCase Ref. P319WO IPTector®prevented data fitting.

[0292] Moving into aqueous solutions, quantitative1H NMR spectroscopy experiments were undertaken in a D2O: EtOH 19:1 solution (2.78 mM), to confirm the presence of higher order selfassociation events, leading to the formation of aggregates which exhibit solid-like properties, and thus appear1H NMR spectroscopy silent. All SSAs and co-formulations exhibited evidence of higher order self-association events under these experimental conditions, see Table 3. Interestingly, for homogeneous solutions of SSAs 6, 33, 39, 74, 75 there was not much variation in the proportion of SSA to be taken up into these higher-order aggregates under these experimental conditions. The dHof these higher-order aggregates, subsequently determined by dynamic light scattering (DLS) for 6, 33, 39, 74, 75 (Table 3), showed those higher order aggregates produced to exhibit a similar size and size distribution profile (intensity peak maxima = 143-184 nm), with the exception of SSA39 (intensity peak maxima = 672 nm), which is the only SSA to contain the inclusion of a benzothiazole moiety.

[0293] Table 4 - Overview of solution state studies of single component homogenous SSAs.DMSO-d6 / 0.5 % H2O H2O: EtOH 1:19SSA CAC ST at CAC Zeta potentialD. (nm) DhK(M'1)h(nm)7PDI dim ' (mM) (mN / M) (mV)6 1.50 2.6 5.61 33.59 -55.70 184.36 0.0315 33 1.42 104.7 b b -55.50 142 0.0350 39 1.64 2.7 0.5 46.5 -86.33 672.27 0.0658 74 1.65 a <7.5 b -79.31 162.46 0.0388 75 1.61 7636 <2.78b b -73.44 174 0.0614

[0294] a - Compound is in slow exchange.

[0295] b - Could not be calculated due to compound solubility.Table 5 - Overview of solution state studies of co-formulated heterogenous mixtures of SSAs.H2O: EtOH 1:19CoSSA ST at Zetaformulation components CACCAC potential Dh(nm) PDI (mM)(mN / M) (mV)a 6 + 33 <10o a 0.99 107.1 0.0745 b 6 + 39 8.06 37.51 -96.99 187.24 0.0600 c 6 + 4 <7.5o a -68.2 107.1 0.0688 d 6 + 5 <2.78a a -53.9 98.7 0.0485 e 33 + 3 2.83 46.38 -71.13 188.8 0.0596 f 33 + 4 <7.5o a -76.06 181.4 0.0374Case Ref. P319WO IPTector®g 33 + 75 <2.78a a -62.2 105.3 0.0103 h 39 + 74 <7.5o a -74.9 189.8 0.0686 i 39 + 75 2.45 42.14 -69.4 689.8 0.0809 j 74 + 75 <2.78a a -82.05 84 0.0580

[0296] 0 - Could not be calculated due to compound solubility.

[0297] The stability of the homogenous aggregates formed in H₂O:EtOH 19:1 at 2.78 mM of 6, 33, 39, 74, 75 were then determined through zeta potential studies. The thiourea based self-associated aggregates were found to be the least stable with zeta potential measurements of -56 mV, substituting the thiourea for the squaramide residue increased the stability of those nanostructures formed, with zeta potential measurements of -79 mV and -73 mV reported for 74 and 75 respectively. We believe that this is due to the increased strength of SSA anion dimerization events (33 Kdim= 105 M1and 75 Kd,m = 7636 M1). Although 3 does not contain a squaramide residue and has a much lower self-associative binding constant of 2.7 M1in DMSO-dg 0.5% H2O, the enhanced stability provided by the planar, hydrophobic benzothiazole functionality gives rise to aggregates with a zeta potential of -86 mV. Tensiometry experiments, the results of which are summarised herein, confirmed 6, 33, 39, 74, 75 to exhibit surfactant properties in a H₂O: EtOH 19:1 solution, as surface tension is lowered upon increasing concentrations of SSAs, however the critical aggregation concentration (CAC) could only be determined for SSAs 6 and 39 due to SSA solubility.

[0298] We then moved to characterize the self-associative properties of two-component heterogeneous SSA systems, co-formulated in 1:1 molecular ratio, giving rise to co-formulations a - j.These co-formulations were prepared in a H2O: EtOH 19:1 solution. Homogenous solutions of each SSA were combined at equal volumes. The heterogeneous solution subsequently underwent an annealing process, in which the solution was heated to 50 °C for 60 seconds, before cooling to room temperature. As summarised herein, the properties of aggregates formed from co-formulations a - j differ quite considerably from the homogenous aggregates of SSA 6, 33, 39, 74, 75, prepared under identical experimental conditions.

[0299] Quantitative1H NMR spectroscopy confirmed the presence of higher-order aggregates with solid-like properties for co-formulations a - j at 2.78 mM, in a 19:1 D2O: EtOH solution. As evidenced within Table 6, the anionic components of a - j were present in ~ 1:1 ratios for all co-formulations.Table 6 - overview of co-formulation anionic / cationic components.AnionCo-formulation Anion 1 Anion 2 Cation(combined)a 52 64 58 64Case Ref. P319WO IPTector®b 66 70 68 69 c a a 59 65 d 52 56 54 67 e 77 73 75 71 f 71 64 68 70 g 71 73 72 70 h 60 49 54 66 i 58 57 58 59 j 33 41 37 49

[0300] However, the heterogenous SSA co-formulations, with the exception of j, demonstrated a greater proportion of SSA to become incorporated into the higher order aggregate when compared to homogenous solutions of 6, 33, 39, 74, 75 at the same total molecular concentration (2.78 mM), under analogous experimental conditions. This demonstrates a cooperative relationship between all SSA combinations with the exception of j, which incorporated the squaramide based SSAs 74 and 75.SSA co-formulation was also found to result in the production of higher-order aggregated species which demonstrated increased variation in hydrodynamic diameter (determined through DLS studies) and stability (determined through zeta potential studies) compared to the homogenous SSA aggregates produced under analogous experimental conditions. In general, the incorporation of 75 led to aggregates with a smaller hydrodynamic diameter (99 nm, 105 nm, 84 nm), unless combined with 39, in which case this trend was reversed as this combination of SSAs lead to the production of the largest heterogenous SSA aggregates identified (hydrodynamic diameter = 690 nm). However, we believe that this size of aggregate may be produced through the aggregation of smaller species present in solution. Zeta potential measurements showed the presence of 39 (increased hydrophobicity and potential to stabilise intermolecular n-n stacking interactions) and 74 (a sulfonate squaramide based SSA) to increase the stability of the heterogenous aggregates to the greatest extent, with the most stable aggregates formed when combining 74 and 75, which we hypothesise to be due to enhanced intermolecular hydrogen bonding strength, provided through the presence of the squaramide moiety. Tensiometry experiments showed that all SSA co-formulations retained the surfactant nature of the homogenous solutions, although solubility prevented calculation of accurate CAC values.Physicochemical properties for additional SSAsTable 7 Overview of the results obtained from qNMR spectroscopy, where the proportion of an SSA to adopt solid-like properties and therefore become NMR silent is confirmed through comparative integration against an internal standard (DCM 1 %), and DLS studies confirm the hydrodynamicCase Ref. P319WO IPTector®diameter (dH) through the reporting of intensity distribution peak maxima, performed in DMSO-dg 1 % DCM and DMSO solutions respectively at 112 mM and 298 K.qNMR(%) duSSAanion cation (nm)139 5 5 <1, 7, c140 25a24ab141 9 6 <1, 67145 15 14 <1, 3, 48146 72a78a<1, 38147 47 47 <1, 100161 50 51 132162 17 17 37,149165 29 28 <1169 0 0 b170 46 43 1171 50 48 2a - Slow exchange means these data should be treated with caution, b - No evidence of stable higher order self-associated structures, c - Wide range of structures dH= 10-100 nm.Table 8 Overview of the results obtained from: qNMR spectroscopy, where the proportion of an SSA to adopt solid-like properties and therefore become NMR silent is confirmed through comparative integration against an internal standard (EtOH 5 %); critical aggregation concentration - CAC (obtained by tensiometry, ST = surface tension); zeta potential; and DLS studies confirm the stability and hydrodynamic diameter (dH) through the reporting of intensity distribution peak maxima. All experiments were performed in D2O or H2O / 5 % EtOH solutions (as appropriate) at 5.56 mM and 298 K, unless stipulated otherwise.qNMR(%)CAC ST at CAC du Zeta potential SSA Anioncation (mM) (mN / m) (nm) (mV) or acid118 63’ j c c 273 -26 120 48' e c c 177 -17 124 64' j c c 199 -22 126 43' e c c 187 -44Case Ref. P319WO IPTector®139 25 20 18.50 35.00 3, 222 -47 140 67 66 12.50 35.00 125 -71 141 88 e 1.45 38.00 116 -71 142 26 23 0.98 37.23 189, 2128 -56 143 19 18 3.05 35.69 107, 753 -43 144 43 51 13.90 32.43 12, 188 -3 145 54 56 4.50 43.00 188b-48 146 53 50 c c 409d-64 147 56 56 0.52 48.00 422 -66 158 13 19 c c 107, 2733 -110 161 0 1 c c 132 -69 162 56 54 82.60 39.94 37 -59 165 f f g g 155h-94h169 14 14 2.00 47.00 186 -82 170 29 28 3.50 46.00 225 -72 171 74 76 1.10 42.00 364b-92 172 8 5 2.25 37.61 2, 189 -55a - Data not collected at the time of original publication, b - Some evidence of higher order structures du > 1000 nm. c - CAC above the limit of solubility, surfactant properties observed, d - Correlation function raises concerns relating to sample stability, e - Signal overlap prevented the reporting of accurate data, f - Studies conducted at 1.0 mM due to sample solubility, accurate data could not be reported, g - No surfactant properties observed to the limit of SSA solubility, h - Studies conducted at 1.5 mM due to SSA solubility, i - Studies conducted at 1.0 mM due to SSA solubility, j - No counter cation present.Table 9 Overview of the results obtained from low-level in silico modelling of the SSA anion. Electrostatic surface potential maxima (Emax) and minima (Emin) values were obtained following energy minimisation using Spartan '24. Kdim values were obtained through the fitting of fluorescence emission (< 0.001 mM) UV-Vis spectroscopy (< 0.001 M) dilution study data, obtained from an H2O / 5 % EtOH solution at 298 K, to both the EK and CoEK self-associative binding isotherm models using Bindfit v0.5.Surface potential KdimSSA (kJ / mol) ± Error (%)(M1)EmaxEmin139 -64 -757 b bCase Ref. P319WO IPTector®140 -163 -1038 b b141 -63 -734 c c145 -48 -735 b b146 -235 -1029 a a147 -48 -715 c c165 -29 -718 3570d0.59d169 -41 -715 d d170 -41 -715 d d171 -20 -699 c ca - Higher order aggregation events hypothesised to exist, data could not be fitted to either the EK or CoEK self-associative isotherm, b = no evidence of molecular association observed, c - Evidence of low order and high order self-association, data could not be fitted to either the EK or CoEK self-associative isotherm, d - Evidence of low order self-association observed, e - Obtained from the fitting of fluorescence emission data, excitation = 331 nm.Table 10 Overview of the results obtained from the SSA anion. cLogP values were obtained using SwissADME.SSA cLogP SSA cLogP SSA cLogP SSA cLogP 1 SSA cLogP 1, 7-13, 0.68 42 0.32 76 3.07 126-127 1.63 162 1.49 872 1.29 43 0.32 77 6.16 130-131 1.91 163 0.59 3 1.54 44 0.52 78 2.62 132-133 1.58 164 1.06 4, 14- 1.44 45 0.77 79 2.62 136 0.97 165 4.94 185 1.91 46 1.86 80 2.78 137 1.07 166 3.90 6 2.24 47 2.48 81 2.78 138 1.30 167 3.18 19, 62 -0.42 48 2.73 82 3.38 139 1.65 168 3.87 20 -0.18 49 1.05 83 3.80 140 1.42 169 2.60 21 -0.16 50 1.51 84 3.34 141 4.23 170 2.60 22 -0.58 51 0.92 85 3.46 142, 0.25 171 4.16173,17623 2.30 52 -0.37 86 1.00 143 -1.21 172, 175, 2.3917824 0.54 53 0.24 88-89 1.46 144, 1.09 179 0.74Case Ref. P319WO IPTector®174,17725 0.31 54 2.24 90-91 1.82 145 1.21 180 1.43 26 -0.50 55 -0.27 92 2.06 146 1.02 183 0.75 27 2.56 56-57 2.93 93 -94 1.70 147 3.70 184 1.57 28 2.99 58-59 3.06 95 -96 1.17 148 3.96 185 2.89 29 1.68 60 2.50 97 -98 1.24 149 0.79 186 5.85 30 1.42 61 2.79 99 - 100, 0.72 150 0.61 187 -0.6311731 3.54 63 -0.18 101 - 102 1.29 151 0.01 188 1.88 32 1.77 65 0.53 103 - 104 0.77 152 0.42 189 2.20 33 1.50 66 -0.38 105 - 106 0.89 153 2.82 190 3.87 34 0.91 67 -0.58 107 - 108 0.40 154 3.1335 1.93 68 0.86 109 - 110 0.65 155 3.8036 2.16 69 0.86 111-112 0.11 156 3.0537 0.80 70 -0.83 113-114 0.25 157 3.5138 1.92 71 0.41 115-116 -0.22 158, 5.18181,18239 2.22 72 -73 2.77 118-119 1.58 159 0.6540 -0.20 74 1.28 120 - 121 1.36 160 0.9141 -0.20 75 0.70 124-125 1.95 161 0.94Example 3 - Anti-microbial application

[0301] The experiments were conducted as described in Example 1, " MIC assay".

[0302] The minimum inhibitory concentration (MIC) values determined for a range of SSAss: 30, 72, 73, 56, 57, and 1:1 enantiomeric mixtures of SSAs 72 + 73 and 56 + 57 against 7 different Gram-positive Staphylococcus aureus (S. aureus), Enterococcus faecalis (E. faecalis) and Enterococcus faecium (E. faecium) bacteria strains. Here, the MIC is the lowest concentration at which no visible growth was observed.MIC(mM)SSA S. aureus S. S. S. aureus E. E. Faecalis E. Faecium 9144 aureus aureus 1199B Faecalis NCTC NCTC 13616 USA 300 NCTC 775 12201 12204 30 1.390 > 2.780 > 2.780 > 2.780 > 2.780 > 2.780 > 2.780 72 0.087 0.087 > 2.780 0.087 > 2.780 > 2.780 > 2.780Case Ref. P319WO IPTector®73 0.174 0.087 > 2.780 2.780 2.780 > 2.780 > 2.780 72+73a0.174 0.087 0.174 0.174 > 2.780 > 2.780 > 2.780 56 0.087 0.087 0.087 0.174 > 2.780 > 2.780 > 2.780 57 0.174 0.087 0.087 0.174 2.780 > 2.780 > 2.780 56+57a0.087 0.087 0.087 0.087 - > 2.780 > 2.780 > 2.7800.349

[0303] Tab e 11 - a- 1:1 mixture.

[0304] Table 12 Overview of MIC (mM, n = 3) results obtained for SSAs against S. aureus (ATCC 9144, NCTC 13616), K. pneumoniae (M6), A baumannii (ATCC 17978), P. aeruginosa (PAO1, NCTC 13437) and E. coli (NCTC 12923) strains.ATCC NCTC ATCC NCTC NCTC SSA M6 PAO19144 13616 17978 13437 12923139 2.500 >5 >5a>5a>5a>5a>5a140 2.500 5.000 >5a>5a>5a>5a>5a141 0.625 0.625 >5a>5a>5a>5a>5a145 1.250 2.500 >5a>5a>5a>5a>5a146 1.250 1.250 >5a>5a>5a>5a>5a147 1.250 0.625 >5a>5a>5a>5a>5a161 0.625 0.625 >5a>5a>5a>5a>5a162 5 >5 >5a>5a>5a>5a>5a165 0.016 0.030 >0.5b>0.5b>0.5b>0.5b>0.5b169 1.250 >5 >5a>5a>5a>5a>5a170 1.250 5.000 >5a>5a>5a>5a>5a171 0.005 0.005 >5a>5a>5a>5a>5a190 0.02 0.02 >2.5a>2.5a>2.5a>2.5a>2.5a

[0305] a - Due to a lack of activity, only two biological repeats were undertaken, b - Solubility prevented testing at higher concentrations.

[0306] Table 13 Overview of MIC results obtained for SSAs in the presence of PMBN (15 pg / mL) against K. pneumoniae (M6), A. baumannii (ATCC 17978), P. aeruginosa (PAO1, NCTC 13437) and E. coli (NCTC 12923) strains.ATCC NCTC NCTC SSA M6 PAO117978 13437 12923139 >5 >5 2.500 5.000 >5Case Ref. P319WO IPTector®140 >5 >5 1.250 2.500 2.500 141 0.300 0.300 0.080 0.300 0.300 145 >5 5.000 5.000 >5 5.000 146 >5 5.000 5.000 >5 2.500 147 0.625 >5 0.040 0.625 0.313 161 2.5 0.6-1.25 0.625 >5 0.6-1.25 162 >5 >5 >5 >5 >5 165 >0.5a>0.5a0.125 >0.5a0.5a169 >5 >5 5.000 >5 >5 170 >5 >5 2.500 >5 >5 171 0.02-0.3 0.010 0.005 >0.5 0.020 190 >2.5 >2.5 0.02 0.02 0.04

[0307] a - Solubility prevented testing at higher concentrations.

[0308] Table 14 Overview of % inhibition at 0.50 mM, n = 3 results obtained for SSAs against S. aureus (ATCC 9144), K. pneumoniae (NCTC 13368), P. aeruginosa (PAO1, NCTC 13437), E.faecalis ( NCTC 775, NCTC 12201), E. Faecium (NCTC 12204).ATCC NCTC NCTC NCTC NCTC NCTC SSA PA019144 13368 13437 775 12201 12204118 24 13 64 18 42 72 81 120 23 40 67 40 44 71 85 124 59 69 96 82 65 75 78 126 34 27 62 20 37 36 62Case Ref. P319WO IPTector®Table 15 - The minimum inhibition concentration of SSAs as TBA, chlorhexidine or octenidine salts against S. aureus (ATCC 9144, NCTC 13616), K. pneumoniae (M6), A baumannii (ATCC 17978), P. aeruginosa (PAO1, NCTC 13437) and E. coli (NCTC 12923) strains. TBA salts (142, 143, 144 and 172) were dissolved in 5 % ethanol and 95 % H₂O. Chlorhexidine (173, 174, 175 and 181) and octenidine (176, 177,178 and 182) salts were dissolved in 10 % DMSO and 90 % H₂O and plated at a final DMSO concentration of 5 %.ATCC NCTC ATCC NCTC NCTC SSA’ M69144 13616 17978 PAO1 13437 12923 142 >2500 >5000 >5000 >5000 >5000 >5000 >5000 143 2500 n / a >5000 >5000 >5000 n / a >5000 144 1250 >5000 >5000 >5000 >5000 >5000 >5000 158 >5000 >5000 >5000 >5000 >5000 >5000 >5000 172 620 >5000 >5000 >5000 >5000 >5000 >5000 173 <0.55 1.11 17.26 17.26 8.85 17.26 <0.55 174 1.07 2.15 67.14 16.76 16.76 33.52 <0.54 175 1.92 3.84 120.21 7.69 30.01 60.11 1.92 176 1.05 1.05 2.11 2.11 4.21 4.21 2.11 177 1.02 1.02 2.05 2.05 4.09 4.09 2.05 178 1.84 0.92 1.84 3.69 3.69 3.69 1.84 181 b b b b b b b 182 <3.30 <3.30 <13.00 <13.00 <37.30 <37.30 <6.50 Chlorhexidine <0.99 1.98 30.86 7.91 7.91 15.83 <0.99 Octenidine1.60 1.60 3.21 3.21 6.41 3.21 1.60 Dihydrochloridea All values are expressed in pM.b Data not availableExample 4 - Anti-cancer application

[0309] The experiments were conducted as described in Example 1, " GI50determination assays".

[0310] The concentration required to reduce / inhibit cellular growth by 50 % (GI50) determined for cisplatin, Tetrabutylammonium (TBA)CI, SSAs 30, 72, 73, 56, 57, and 1:1 enantiomeric mixtures of 72 + 73 and 56 + 57 against non-cancerous RPE-1 cells, ovarian cancer cells (A2780) and cisplatin resistant ovarian cancer cells (A2780 CisR).RPE-1 SRB Assays A2780 SRB Assays A2780 CisR SRB Assays SSA Mean SD 2 2 2R >0.9 Mean GI50SD R2> Mean GI50SD + / - R2GI50(µM) + / - n =(pM) + / - 0.9 >(pM)n = 0.9 n = Cisplatin 6.25 1.44 3 1.088 0.35 8 10.804 1.77 4Case Ref. P319WO IPTector®TBACI 532.46 222 3 114.16 4.01 3 219.26 11.21 3 30 295.93 210 3 116.53 6.50 3 367.20 62.46 3 72 > 500 b b 171.03 12.45 3 276.06 39.7 3 73 > 375 b b 230.90 9.25 3 498.40 50.17 3 72+73a> 375 b b 162.40 21.90 3 334.40 91.00 3 56 > 375 b b 177.76 16.47 3 356.33 36.94 3 57 > 375 b b 104.93 4.44 3 196.72 9.25 4 56+57a> 375 b b 86.81 0.90 3 160.33 5.35 3Table 16 - a - 1:1 mixture, b = not applicable.Example 5 - Synthesis of SSA's and precursors

[0311] TBA = tetrabutylammonium.

[0312] SSA30: This was synthesised in line with previously published methods (L. J. White, S. N. Tyuleva, B. Wilson, H. J. Shepherd, K. K. L. Ng, S. J. Holder, E. R. Clark and J. R. Hiscock, Chem. Eur. J., 2018, 24, 7761-7773).1H NMR (400 MHz, 298 K, DMSO-d6): 6: 10.27 (s, 1H), 7.68 (d, J = 8.60 Hz, 2H), 7.50 (d, J = 8.60 Hz, 2H), 6.75 (s, 1H), 3.18 - 3.13 (m, 8H), 1.60 - 1.52 (m, 8H), 1.34 - 1.25 (m, 8H), 0.93 (t, J = 14.64 Hz, 12H). F3CO intermediate 1

[0313] Intermediate 1: l-lsocyanato-4-(trifluoromethyl) benzene (0.29 mL, 2.00 mmol) was added to L-Leucine tert-butyl ester hydrochloride (0.45 g, 2 mmol) in anhydrous pyridine (10 mL) and stirred at room temperature overnight. The mixture was then taken to complete dryness and redissolved in chloroform (5 mL), followed by dropwise additions of hexane (> 5 mL) resulting in precipitation. The precipitate was collected via filtration and purified via flash column chromatography with a 3:2 mixture of hexane to ethyl acetate. All fractions were monitored using thin layer chromatography and were taken to complete dryness. The pure product was identified using NMR spectroscopy and collected as a white solid with a yield of 53 % (0.40 g, 1.07 mmol); melting point: > 200 °C;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 8.96 (s, 1H), 7.57 (s, 4H), 6.55 (d, J = 8.12 Hz, 1H), 4.13 (q, J = 22.92 Hz, 1H), 1.71 - 1.62 (m, 1 H), 1.54 - 1.48 (m, 2H), 1.41 (s, 9H), 0.91 (q, J = 18.80 Hz, 6H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 172.8 (C=O), 154.9 (C=O), 144.3 (ArC), 126.5 (q, J = 3.81 Hz, ArCH), 123.7Case Ref. P319WO IPTector®(ArC), 121.7 (q, J = 31.78 Hz, CF3), 117.7 (ArCH), 81.1 (C), 51.9 (CH), 41.3 (CH2), 28.1 (CH), 24.9 (CH3), 23.1 (CH3), 22.1 (CH3); IR (film): v = 3338 (NH stretch), 2958, 1701, 1680, 1527, 1332, 1315, 1155, 1105, 1066, 833, 607; HRMS for the carboxylate-urea ion (Ci4Hi6F3N2O3); HRMS for the tert-butyl carboxylate-urea (CI8H25F3N2O3) (ESI ): m / z: act = 374.1739 [M ] ’, cal = 374.1817 [M]’.

[0314] SSA72: Zinc bromide (2.25 g, 10 mmol) was added to intermediate 1 (0.40 g, 1.07 mmol) in dichloromethane (5 mL) and stirred at room temperature for 24 hours. The reaction mixture was then quenched with water (20 mL) and stirred at room temperature for 4 hours, resulting in precipitation. The precipitate was collected via filtration and TBA hydroxide in methanol (0.51 mL, 1 M) was added. The mixture was taken to dryness, and the pure product was collected via flash column chromatography with 100 % ethyl acetate followed by 100 % methanol. The methanol fraction was taken to dryness to give the pure product as a brown solid with a yield of 62 % (0.25 g, 0.45 mmol); melting point: 131 °C;3H NMR (400 MHz, 298 K, DMSO-d6): 6: 9.85 (s, 1H), 7.65 (d, J = 8.48 Hz, 2H), 7.49 (d, J = 8.40 Hz, 2H), 6.74 (s, 1H), 3.74 (q, J = 18.80 Hz, 1H), 3.19 - 3.14 (m, 8H), 1.74 - 1.67 (m, 1H), 1.61 - 1.48 (m, 9H), 1.40 - 1.24 (m, 9H), 0.96 - 0.84 (m, 18H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 174.9 (C=O), 155.4 (C=O), 146.3 (ArC), 125.9 (d, J = 3.84 Hz, ArCH), 124.0 (ArC), 119.9 (q, J = 31.44 Hz, CF3), 117.2 (ArCH), 58.0 (CH2), 54.2 (CH), 44.6 (CH2), 25.0 (CH3), 23.7 (CH3), 23.5 (CH2), 23.4 (CH3), 19.6 (CH2), 13.9 (CH3); IR (film): v = 3311 (NH stretch), 2960, 1697, 1604, 1319, 1105, 1064, 842; HRMS for the carboxylate-urea ion (Ci4Hi6F3N2O3(ESI ): m / z: act = 317.1110 [M]’, cal = 317.1118 [M]-.

[0315] Intermediate 2: l-lsocyanato-4-(trifluoromethyl) benzene (0.29 mL, 2.00 mmol) was added to D-Leucine tert-butyl ester hydrochloride (0.45 g, 2 mmol) in anhydrous pyridine (10 mL) and stirred at room temperature overnight. The mixture was then taken to complete dryness and redissolved in chloroform (5 mL), followed by dropwise additions of hexane (> 5 mL) resulting in precipitation. The precipitate was collected via filtration and purified via flash column chromatography with a 3:2 mixture of hexane to ethyl acetate. All fractions were monitored using thin layer chromatography and were taken to complete dryness. The pure product was identified using NMR spectroscopy and collected as a white solid with a yield of 75 % (0.56 g, 1.50 mmol); melting point: > 200 °C;3H NMRCase Ref. P319WO IPTector®(400 MHz, 298 K, DMSO-d6): δ: 8.96 (s, 1H), 7.57 (s, 4H), 6.55 (d, J = 8.12 Hz, 1H), 4.13 (q, J = 22.76 Hz, 1H), 1.70 - 1.62 (m, 1 H), 1.52 - 1.48 (m, 2H), 1.41 (s, 9H), 0.91 (q, J = 18.72 Hz, 6H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 172.8 (C=O), 154.9 (C=O), 144.3 (ArC), 126.5 (q, J = 3.82 Hz, ArCH), 123.7 (ArC), 121.7 (q, J = 31.78 Hz, CF3), 117.7 (ArCH), 81.1 (C), 51.9 (CH), 41.3 (CH2), 28.1 (CH3), 24.9 (CH3), 23.1 (CH3), 22.1 (CH3); IR (film): v = 3338 (NH stretch) 2958, 1703, 1680, 1529, 1332, 1315, 1155, 1107, 1066, 833, 609; HRMS for the tert-butyl carboxylate-urea (CigH25F3N2O3) (ESI ): m / z: act = 374.1763 [M]“, cal = 374.1817 [M]\SSA73

[0316] SSA73: Zinc bromide (2.25 g, 10 mmol) was added to intermediate 2 (0.56 g, 1.50 mmol) in dichloromethane (5 mL) and stirred at room temperature for 24 hours. The reaction mixture was then quenched with water (20 mL) and t stirred at room temperature for 4 hours, resulting in precipitation. The precipitate was collected via filtration and TBA hydroxide in methanol (0.51 mL, IM) was added. The mixture was taken to dryness, and the pure product was collected via flash column chromatography with 100 % ethyl acetate followed by 100 % methanol. The methanol fraction was taken to dryness to give the pure product as a brown solid with a yield of 68 % (0.38 g, 0.68 mmol); melting point: 120 °C;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 10.01 (s, 1H), 7.67 (d, J = 8.28 Hz, 2H), 7.49 (d, J = 8.60 Hz, 2H), 6.81 (s, 1H), 3.75 (q, J = 18.64 Hz, 1H), 3.19 - 3.14 (m, 8H), 1.75 - 1.66 (m, 1H), 1.61 - 1.48 (m, 9H), 1.41 - 1.24 (m, 9H), 0.96 - 0.85 (m, 18H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 175.0 (C=O), 155.4 (C=O), 146.3 (ArC), 125.9 (d, J = 3.9 Hz, ArCH), 121.7 (q, J = 31.48 Hz, CF3), 117.2 (ArCH), 58.0 (CH2), 54.2 (CH), 44.5 (CH2), 25.0 (CH3), 23.7 (CH3), 23.5 (CH2), 23.4 (CH3), 19.6 (CH2), 13.9 (CH3); IR (film): v = 3309 (NH stretch), 2960, 1697, 1606, 1319, 1107, 1066, 846; HRMS for the carboxylate-urea ion (Ci4Hi6F3N2O3) (ESI ): m / z: act = 317.1160 [M]’, cal = 317.1118 [M]’.O N A NH Hintermediate 3

[0317] Intermediate 3: l-lsocyanato-4-(trifluoromethyl) benzene (0.29 mL, 2.00 mmol) was added to a stirring solution of L-Phenylalanine tert-butyl ester hydrochloride (0.52 g, 2 mmol) in anhydrous pyridine (10 mL) and left at room temperature overnight. The mixture was then taken to complete dryness and redissolved in chloroform (5 mL), followed by dropwise additions of hexane (> 5 mL) resulting in precipitation. The precipitate was collected via filtration and purified via flash column chromatography with a 3:2 mixture of hexane to ethyl acetate. All fractions were monitored using thinCase Ref. P319WO IPTector®layer chromatography and were taken to complete dryness. The pure product was identified using NMR spectroscopy and collected as a white solid with a yield of 61 % (0.50 g, 1.22 mmol); melting point: > 200 °C;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 9.10 (s, 1H), 7.56 (s, 4H), 7.33 - 7.20 (m, 5H), 6.52 (d, J = 7.52 Hz, 1H), 4.41 (q, J = 20.64 Hz, 1H), 2.99 (d, J = 5.04 Hz, 2H), 1.35 (s, 9H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 171.5 (C=O), 154.7 (C=O), 144.2 (ArC), 137.2 (ArC), 129.8 (ArCH), 128.7 (ArCH), 127.0 (ArCH), 126.4 (q, J = 3.77 Hz, ArCH), 121.5 (q, J = 31.75 Hz, CF3), 123.7 (ArC), 117.7 (ArCH), 81.4 (C), 54.6 (CH), 37.9 (CH2), 28.0 (CH3); IR (film): v = 3381 (NH stretch), 2987, 1681, 1535, 1327, 1315, 1234, 1159, 1107, 1066, 835, 700, 609; HRMS for the tert-butyl carboxylate-urea (C2IH23F3N2O3) (ESI ): m / z: act = 408.1227 [M] cal = 408.1661 [M]’.SSA56

[0318] SSA56: Zinc bromide (2.25 g, 10 mmol) was added to a stirring solution of intermediate 3 (0.50 g, 1.22 mmol) in dichloromethane (5 mL) and stirred at room temperature for 24 hours. The reaction mixture was then quenched with water (20 mL) and stirred at room temperature for 4 hours, resulting in precipitation. The precipitate was collected via filtration and TBA hydroxide in methanol (0.51 mL, IM) was added. The mixture was taken to dryness, and the pure product was collected via flash column chromatography with 100 % ethyl acetate followed by 100 % methanol. The methanol fraction was taken to dryness to give the pure product as a brown solid with a yield of 54 % (0.27 g, 0.45 mmol); melting point: 160 °C;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 9.81 (s, 1H), 7.64 (d, J = 8.64 Hz, 2H), 7.49 (d, J = 8.64 Hz, 2H), 7.15 - 7.04 (m, 5H), 6.61 (d, J = 5.12 Hz, 1H), 3.96 (q, J = 15.92 Hz, 1H), 3.17 -2.97 (m, 10 H), 1.60 - 1.51 (m, 8H), 1.34 - 1.25 (m, 8H), 0.92 (t, J = 14.64 Hz, 12H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 173.2 (C=O), 155.2 (C=O), 146.0 (ArC), 140.3 (ArC), 130.1 (ArCH), 128.0 (ArCH), 126.0 (q, J = 3.75 Hz, ArCH), 125.8 (ArCH), 124.0 (ArC), 120.2 (q, J = 31.55 Hz, CF3), 117.2 (ArCH), 57.9 (t, J = 2.5 Hz, CH2), 56.7 (CH), 39.1 (CH2), 23.5 (CH2), 19.7 (CH2), 13.9 (CH3); IR (film): v = 3419 (NH stretch), 2964, 1695, 1591, 1485, 1394, 1319, 1109, 1064, 846, 705; HRMS for the carboxylate-urea ion (CI7HI4F3N2O3) (ESI ): m / z: act = 351.0952 [M]’, cal = 351.0962 [M]’.intermediate 4

[0319] Intermediate 4: l-lsocyanato-4-(trifluoromethyl) benzene (0.29 mL, 2.00 mmol) was added to D-phenylalanine tert-butyl ester hydrochloride (0.52 g, 2 mmol) in anhydrous pyridine (10 mL) and stirred at room temperature overnight. The mixture was then taken to complete dryness andCase Ref. P319WO IPTector®redissolved in chloroform (5 mL), followed by dropwise additions of hexane (> 5 mL) resulting in precipitation. The precipitate was collected via filtration and purified via flash column chromatography with a 3:2 mixture of hexane to ethyl acetate. All fractions were monitored using thin layer chromatography and were taken to complete dryness. The pure product was identified using NMR spectroscopy and collected as a white solid with a yield of 62 % (0.51 g, 1.25 mmol); melting point: 166 °C;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 9.10 (s, 1H), 7.56 (s, 4H), 7.33 - 7.21 (m, 5H), 6.53 (d, J = 8.12 Hz, 1H), 4.41, (q, J = 21.40 Hz, 1H), 3.00 (d, J = 6.88 Hz, 2H), 1.35 (s, 9H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 171.5 (C=O), 154.7 (C=O), 144.3 (ArC), 137.3 (ArC), 129.8 (ArCH), 128.7 (ArCH), 127.1 (ArCH), 126.5 (q, J = 3.79 Hz, ArCH), 121.8 (q, J = 31.78 Hz, CF3), 123.7 (ArC), 117.7 (ArCH), 81.5 (C), 54.6 (CH), 38.0 (CH2), 28.0 (CH3); IR (film): v = 3381 (NH stretch), 2985, 1681, 1537, 1328, 1317, 1236, 1159, 1107, 1066, 835, 700, 609; HRMS forthe tert-butyl carboxylate-urea (C2IH23F3N2O3) (ESI ): m / z: act: 408.1502 [M]’ cal: 408.1661 [M].

[0320] SSA57: Zinc bromide (2.25 g, 10 mmol) was added to intermediate 4 (0.51 g, 1.25 mmol) in dichloromethane (5 mL) and stirred at room temperature for 24 hours. The reaction mixture was then quenched with water (20 mL) and stirred at room temperature for 4 hours, resulting in precipitation. The precipitate was collected via filtration and TBA hydroxide in methanol (0.51 mL, IM) was added. The mixture was taken to dryness, and the pure product was collected via flash column chromatography with 100 % ethyl acetate followed by 100 % methanol. The methanol fraction was taken to dryness to give the pure product as a brown solid with a yield of 54 % (0.28 g, 0.47 mmol); melting point: > 200 °C;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 9.71 (s, 1H), 7.63 (d, J = 8.60 Hz, 2H), 7.50 (d, J = 8.72 Hz, 2H), 7.31 - 7.05 (m, 5H), 6.57 (s, 1H), 3.87 (q, J = 14.44 Hz, 1H), 3.18 - 2.98 (m, 10 H), 1.60 - 1.51 (m, 8H), 1.35 - 1.25 (m, 8H), 0.93 (t, J = 14.88 Hz, 12H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 173.2 (C=O), 155.2 (C=O), 146.0 (ArC), 140.2 (ArC), 130.1 (ArCH), 128.0 (ArCH), 126.0 (q, J = 3.62 Hz, ArCH), 125.8 (ArCH), 124.0 (ArC), 120.2 (q, J = 31.54 Hz, CF3), 117.3 (ArCH), 58.0 (t, J = 2.46 Hz, CH2), 56.7 (CH), 39.1 (CH2), 23.5 (CH2), 19.7 (CH2), 13.9 (CH3); IR (film): v = 3309 (NH stretch), 2962, 1695, 1606, 1489, 1381, 1319, 1107, 1066, 846, 702; HRMS for the carboxylate-urea ion (CI7HI4F3N2O3- ) (ESI ): m / z: act: 351.0937 [M]’ cal: 351.0962 [M]’.Case Ref. P319WO IPTector®

[0321] SSA6: Aminopropansulfonic acid (0.278 g, 2.00 mmol) was dissolved in tetrabutylammonium in methanol (2.00 mL, 2 mmol) with excess methanol (15 mL) to aid dissolving before being taken to complete dryness. Tetrabutylammonium aminopropansulfonate (2.00 mmol) was then dissolved in ethyl acetate (20 mL) and l-isothiocyanato-4-(trifluoromethyl)benzene (0.406 g, 2.00 mmol) was added and stirred overnight at room temperature. The crude precipitate was filtered and rinsed with ethyl acetate (5 mL) resulting in the pure product as a white solid (0.728 g, 1.25 mmol, 63.3 %);1H NMR (400 MHz, 298 K, DMSO-d6): 0.93 (t, J = 7.28 Hz, 12H), 1.26 - 1.35 (m, 8H), 1.53 - 1.60 (m, 8H), 1.81 - 1.88 (m, 2H), 2.48 (s, 2H), 3.14 - 3.18 (m, 8H), 3.56 (d, J = 3.92 Hz, 2H), 7.62 (d, J = 8.52 Hz, 2H), 7.76 (d, J = 7.32 Hz, 2H), 8.19 (s, 1H), 9.89 (s, 1H).I BAO SSA33

[0322] SSA33: Glycine (0.150 g, 2.00 mmol) was dissolved in dH2O (5 mL) and tetrabutylammonium hydroxide (2.00 mL, 2.00 mmol) was added. The resultant mixture was taken to dryness, with the aid of toluene (10 mL). Glcyinate tetrabutylammonium was then dissolved in ethyl acetate (20 mL) and 1-isothiocyanato-4-(trifluoromethyl)benzene (0.406 g, 2.00 mmol) was added and stirred overnight at room temperature. The crude precipitate was filtered and rinsed with ethyl acetate (5 mL) resulting in the pure product as a white solid (0.670 g, 1.29 mmol, 64.5 %);1H NMR (400 MHz, 298 K, DMSO-d6) 0.92 (t, J = 7.28 Hz, 12H), 1.25 - 1.35 (m, 8H), 1.52 - 1.59 (m, 8H), 3.13 - 3.17 (m, 8H), 3.66 (s, 2H), 7.57 (d, J = 8.56 Hz, 2H), 8.20 (d, 8.28 Hz, 2H), 8.49 (s, 1H), 11.66 (s, 1H).

[0323] SSA39: Aminomethansulfonic acid (0.222 g, 2.00 mmol) was dissolved in tetrabutylammonium in methanol (2.00 mL, 2 mmol) with excess methanol (15 mL) to aid dissolving before being taken to complete dryness. Tetrabutylammonium aminomethansulfonate (2.00 mmol) was then dissolved in ethyl acetate (20 mL), 4-(6-methylbenzo[d]thiazol-2-yl)aniline (0.480 g, 2 mmol) was added and theCase Ref. P319WO IPTector®mixture was stirred at room temperature for 24 hours. The crude precipitate was filtered and rinsed with ethyl acetate (5 mL), then dissolved in chloroform (15 mL) and washed with water (15 mL x 3), and taken to dryness. Ethyl acetate (20 mL) was then added to the organic oil and sonicated for 30 minutes, resulting in a precipitate which was filtered and washed with ethyl acetate (5 mL) resulting in the pure product as a white solid (0.974 g, 1.57 mmol, 78.6 %).1H NMR (400 MHz, 298 K, DMSO-d6) 0.93 (t, J = 7.28 Hz, 12H), 1.25 - 1.35 (m, 8H), 1.52 - 1.59 (m, 8H), 2.44 (s, 3H), 3.13 - 3.17 (m, 8H), 3.92 (d, J = 5.80 Hz, 2H), 6.75 (s, 1H), 7.31 (d, J = 8.36 Hz, 1H), 7.56 (d, J = 8.72 Hz, 2H), 7.85 - 7.92 (m, 4H), 9.18 (s, 1H).SSA74

[0324] SSA74: Aminopropansulfonic acid (0.278 g, 2.00 mmol) was dissolved in tetrabutylammonium in methanol (2.00 mL, 2 mmol) with excess methanol (15 mL) to aid dissolving before being taken to complete dryness. Tetrabutylammonium aminopropansulfonate (2.00 mmol) was then dissolved in ethyl acetate (20 mL), 3-methoxy-4-((4-(trifluoromethyl)phenyl)amino)cyclobutane-l,2-dione(0.271 g, 2 mmol) was added and the mixture was stirred at room temperature for 24 hours.The crude precipitate was filtered and rinsed with ethyl acetate (5 mL) resulting in the pure product as a white solid (0.906 g, 1.46 mmol, 87 %); melting point: 161 °C;1H NMR (400 MHz, 298 K, DMSO-d6) 0.92 (t, 12H, J = 7.40 Hz), 1.25 - 1.34 (m, 8H), 1.52 - 1.60 (m, 8H), 1.94 (t, 2H, J = 6.88 Hz), 2.66 (t, 2H, 7.64 Hz), 3.13 - 3.18 (m, 8H), 3.72 (s, 2H), 7.63 - 7.72 (m, 4H), 8.14 (s, 0.8H), 8.71 (s, 0.1H), 10.23 (s, 1H);13C{1H} NMR (400 MHz, 298 K, DMSO-d6): 13.3 (CH3), 19.1 (CH2), 22.9 (CH2), 26.7 (CH2), 42.6 (CH2), 48.0 (CH2), 57.4 (CH2), 117.9 (ArCH), 122.1 (q, Ji = 128.0 Hz, ArC), 124.4 (q, Ji = 1096.0 Hz, CF3), 126.3 (q, Ji = 12.0 Hz, ArCH), 142.8 (ArC), 163.0 (ArC), 169.6 (ArC), 179.9 C=O, 184.5 C=O; IR (film): v = 1788 (C=O stretch), 1690, 1612, 1589, 1420, 1034.SSA75

[0325] SSA75: Glycine (0.150 g, 2.00 mmol) was dissolved in dH2O (5 mL) and tetrabutylammonium hydroxide (2.00 mL, 2.00 mmol) was added. The resultant mixture was taken to dryness, with the aid of toluene (10 mL). Glcyinate tetrabutylammonium was then dissolved in ethyl acetate (20 mL) and 3-methoxy-4-((4-(trifluoromethyl)phenyl)amino)cyclobutane-l, 2-dione (0.271 g, 2 mmol) was addedCase Ref. P319WO IPTector®and the mixture was stirred at room temperature for 24 hours. The crude precipitate was filtered and rinsed with ethyl acetate (5 mL) resulting in the pure product as a white solid. (0.523 g, 0.942 mmol, 47 %); melting point: <200 °C;1H NMR (400 MHz, 298 K, DMSO-d6): 0.91 (t, J = 7.28 Hz, 12H), 1.24 -1.33 (m, 8H), 1.51 - 1.59 (m, 8H), 3.12 - 3.17 (m, 8H), 4.13 (s, 2H), 7.58 (d, J = 8.72 Hz, 2H), 8.28 (d, J = 8.44 Hz, 2H), 8.96 (s, 1H), 13.0 (s, 1H);13C{1H} NMR (400 MHz, 298 K, DMSO-d6): 13.4 (CH3), 19.2 (CH2), 23.0 (CH2), 48.1 (CH2), 57.5 (CH2), 118.8 (ArCH), 121.5 (q, Ji = 128.3 Hz, ArC), 124.7 (q, Ji = 1077.2 Hz, CF3), 126.0 (q, Ji = 14.6 Hz, ArCH), 144.2 (ArC), 163.5 (ArC), 168.8 (ArC), 170.5 (C=O), 180.4 (C=O), 185.1 (C=O); IR (film): v = 3219, 1788, (C=O stretch), 1680, 1636, 1589, 1437, 1107;

[0326] 3-methoxy-4-((4-(trifluoromethyl)phenyl)amino)cyclobutane-l, 2-dione: A mixture of 4-(trifluoromethyl)analine (0.88 mL, 7.00 mmol) and 3, 4-dimethoxycyclobut-3-ene-l, 2-dione was stirred at room temperature in MeOH (15 mL) for 48 hours. The pure product was isolated by filtration as a pale yellow solid (1.24 g, 5.25 mmol, 75.2 %).1H NMR (400 MHz, 298 K, DMSO-d6): 4.40 (s, 3H), 7.55 (d, J = 8.44 Hz, 2H), 7.71 (d, J = 8.52 Hz, 2H) 11.01 (s, 1H).L118 / D119

[0327] SSA 118: N, N'-Carbonyldiimidazole (5.60 mmol, 0.91 g) was added to 4-methylbenzenesulfonamide (4.00 mmol, 0.69 g) in dichloromethane (30 mL) and refluxed for 4 hours. At the same time triethylamine (5.20 mmol, 0.73 mL) was added to L-Leucine methyl ester hydrochloride (4.80 mmol, 0.87 g) in dichloromethane (10 mL) and stirred at room temperature. After 4 hours the L-leucine solution was added to the refluxing solution, and the resulting solution was refluxed overnight. The mixture was taken to complete dryness and sonicated in ethyl acetate (20 mL) until white precipitate formed. The precipitate was collected through filtration and was dissolved in 1:1 methanokIPA (20 mL) and 4M NaOH (4.00 mmol, 1.00 mL) was added. The mixture was stirred overnight at room temperature before being acidified using cone. HCI to pH 2-3. The solution was concentrated under reduced pressure until a white precipitate formed. The precipitate was collected through filtration, and the pure product collected as a white solid with a yield of (1.20 g, 3.94 mmol, 92 %): Melting point: > 473K;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 0.80 (dd, J = 27.4 Hz, 5.6 Hz, 6H), 1.46 (s, 3H), 2.39 (s, 3H), 4.01 (s, 1H), 6.68 (m, J = 8.08 Hz, 1H), 7.40 (d, J = 8.12 Hz, 2H), 7.77 (d, J = 8.28 Hz, 2H), 10.53 (s, 1H), 12.77 (s, 1H).13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 21.0 (CH3), 21.5 (CH), 22.6 (CH), 24.2 (CH), 40.4 (CH2), 50.8 (CH), 127.7 (ArCH), 129.4 (ArCH), 137.7 (ArC), 143.7 (ArC), 151.0 (CO), 173.6 (CO). IR (film): vmax(cm1) = 3229 (NH stretch), 2873, 1643, 1549, 1487, 1257, 1153, 877; HRMS (ESI-): m / z measured = 327.1007 [M]’, predicted = 327.1093 [M]’.Case Ref. P319WO IPTector®

[0328] SSA 120: SSA 118 was dissolved in methanol (10 mL) and tetraethylammonium hydroxide (IM, 4 mL) was added. The mixture was taken to complete dryness, and the pure product was collected as a white solid with a yield of (1.80 g, 3.94 mmol, 100 %). Melting point: > 473 K;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 0.82 (dd, J = 32.04 Hz, 19.56, 6H), 1.17 (m, 12H), 1.47 (m, 3H), 2.39 (s, 3H), 3.21 (m, 8H), 3.98 (m, 1H), 7.17 (d, J = 7.92 Hz, 1H), 7.40 (d, J = 8.04, 2H), 7.78 (d, J = 8.32 Hz, 2H), 10.95 (s, 1H).13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 7.6 (CH3), 21.5 (CH3), 21.9 (CH), 23.1 (CH), 24.7 (CH2), 40.8 (CH2), 49.0 (CH), 51.3 (CH), 51.9 (CH2), 127.7 (ArCH), 129.8 (ArCH), 137.8 (ArC), 144.0 (ArC), 151.8 (CO), 174.1 (CO). IR (film): vmax(cm1) = 3242 (NH stretch), 2883, 1543, 1541, 1487, 1157, 1153.L124 / D125

[0329] SSA 124: N, N'-Carbonyldiimidazole (5.60 mmol, 0.91 g) was added to 4-methylbenzenesulfonamide (4.00 mmol, 0.69 g) in dichloromethane (30 mL) and refluxed for 4 hours. At the same time triethylamine (5.20 mmol, 0.73 mL) was added to L-Phenylalanine methyl ester hydrochloride (4.80 mmol, 1.03 g) in dichloromethane and stirred at room temperature. After 4 hours the L-Phenylalanine solution was added to the refluxing solution, and the resulting solution was refluxed overnight. The mixture was taken to complete dryness and redissolved in dichloromethane (30 mL) and washed with IM HCI (2 x 30 mL) then brine (30 mL). The organic layer was dried, redissolved in methanol (< 5 mL) and precipitated using water. The precipitate was collected through filtration and was dissolved in 1:1 methanokIPA (20 mL) and 4M NaOH (4.00 mmol, 1.00 mL) was added. The mixture was stirred overnight at room temperature before being acidified using cone. HCI to pH 2-3. The solution was concentrated under reduced pressure until a white precipitate formed. The precipitate was collected through filtration, and the pure product collected as a white solid with a yield of (1.02 g, 3.69 mmol, 70 %). Melting point: > 473K;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 2.40 (s, 3H), 2.96 (qd, J = 61.44 Hz, 21.52 Hz, 2H), 4.29 (m, 1H), 6.62 (s, 1H), 7.03 (d, J = 7.40 Hz, 2H), 7.23 (m, 3H), 7.40 (d, J = 8.12 Hz, 2H), 7.73 (d, J = 8.24 Hz, 2H), 10.58 (s, 1H), 12.99 (s, 1H).13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 21.1 (CH3), 36.6 (CH2), 53.5 (CH), 126.6 (ArCH), 127.2 (ArCH), 128.3 (ArCH), 129.2 (ArCH), 129.5 (ArCH), 136.5 (ArC), 137.1 (ArC), 143.8 (ArC), 150.8 (ArCO), 172.3 (ArCO). Vmax (cm’1) = 3334 (NH stretch), 2877, 1641, 1542, 1447, 1439; HRMS (ESI- ): HRMS (ESI-): m / z measured = 361.0850 [M]’, predicted = 362.0936 [M]’.Case Ref. P319WO IPTector®L126 / D127

[0330] SSA 126: SSA 124 was dissolved in methanol (10 mL) and tetraethylammonium hydroxide (IM, 4 mL) was added. The mixture was taken to complete dryness, and the pure product was collected as a white solid with a yield of (1.80 g, 3.94 mmol, 100%). Melting point: > 473K;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 1.15 (m, 12H), 2.38 (s, 3H), 2.93 (s, 2H), 3.20 (q, J = 21.76 Hz, 7.28 Hz, 8H), 4.25 (s, 1H), 6.75 (s, 1H), 7.06 (s, 2H), 7.22 (d, J = 5.92 Hz, 3H), 7.37 (d, J = 7.48 Hz, 2H), 7.71 (d, J = 8.16 Hz, 2H), 10.73 (s, 1H).13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 7.6 (CH3), 21.5 (CH3), 37.1 (CH2), 51.9 (CH2), 54.1 (CH), 127.1 (ArCH), 127.6 (ArCH), 128.7 (ArCH), 129.7 (ArC), 129.9 (ArC), 137.1 (ArC), 137.6 (ArC), 144.1 (CO), 151.5 (CO). IR (film): vmax(cm-1) = 3224 (NH stretch), 2877, 1638, 1552, 1481, 1257, 1153, 878.

[0331] SSA 139: Glycine (0.99 g, 13.10 mmol) was dissolved in methanol (10.00 mL) and tetrabutylammonium hydroxide in methanol (14.00 mL, IM) added. The sample was dried for 24 hours under reduced pressure to give a clear oil (4.12 g, 13.05 mmol). The tetrabutylammonium salt was then dissolved in ethyl acetate (30.00 mL) and an inert atmosphere was then applied to it. This brownoil was then added to a solution of 4-butylphenylisocyonate (2.12 mL, 12.00 mmol) in dichloromethane (30.00 mL) dropwise and the mixture was then heated to 60 °C. After 48 hours the solvent was removed under reduced pressure to give a crude product as a white solid (6.41 g, 13.48 mmol). Purification by silica gel chromatography (100 % ethyl acetate followed by 100 % methanol) afforded the pure product as a white precipitate (1.86 g, 3.91 mmol, 28 %). Melting point 396 K;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 0.89 (t, J = 7.4 Hz, 3H), 0.94 (t, J = 7.4 Hz, 12H), 1.25-1.38 (m, 10H), 1.45-1.64 (m, 10H), 2.46 (t, J = 7.7 Hz, 2H), 3.17 (t, J = 8.5 Hz, 8H), 3.26 (d, J = 3.7 Hz, 2H), 6.27 (d, J = 2.9 Hz, 1H), 6.98 (d, J = 8.4 Hz, 2H), 7.32 (d, J = 8.4 Hz, 2H), 9.18 (s, 1H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 13.5 (CH3), 13.9 (CH3), 19.2 (CH2), 21.8 (CH2), 23.1 (CH2), 33.5 (CH2), 34.2 (CH2), 45.1 (CH2), 57.5 (CH2), 117.2 (ArCH), 128.2 (ArCH), 133.7 (ArC), 139.3 (ArC), 155.2 (C=O), 170.3 (C=O); IR (film): vmax(cm-1) = 3438, 3331 (NH stretch), 2958, 2928, 2873, 1691, 1599, 1545, 1415, 1488; HRMS for the carboxylate-urea ion (C13H17N2O3) (ESI ): m / z measured = 249.1229 [M]’, predicted = 249.1244 [MT.Case Ref. P319WO IPTector®140

[0332] SSA 140: D-glutamic acid (1.93 g, 13.10 mmol) was dissolved in methanol (10.00 mL) and tetrabutylammonium hydroxide in methanol (28.00 mL, 1.00 M) added. The solvent was removed under reduced pressure and further dried for 24 hours under reduced pressure to give a yellow oil (8.10 g, 12.86 mmol). The tetrabutylammonium salt was then dissolved in anhydrous dichloromethane (30.00 mL). Once stirring, 4-butylphenylisocyonate (2.12 mL, 12.00 mmol) was added dropwise, and the mixture taken to 60 °C temperature. After 48 hours the solvent was removed under reduced pressure. The crude product was then dissolved in chloroform (30.00 mL) and then flooded with hexane to induce crystallisation of the partially pure product as a sticky yellow oil. Purification by column chromatography on silica gel using a gradient of hexane / ethyl acetate (10-40 % hexane) afforded the pure compound as a sticky oil of a deep orange colour (0.95 g, 1.18 mmol, 10 %). Melting point - 340 K;TH NMR (400 MHz, 298 K, CD3OD): 6: 0.89 (t, J = 7.3 Hz, 3H), 0.95 (t, J = 7.4 Hz, 24H), 1.27-1.41 (m, 18H), 1.48-1.66 (m, 20H), 1.88-2.16 (m, 3H), 2.47 (d, J = 7.68 Hz, 1H), 3.16-3.22 (m, 18H), 5.96 (s, 1H), 6.94 (d, J = 8.3 Hz, 2H), 7.50 (d, J = 7.2 Hz, 2H), 10.35 (s, 1H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 4.8 (CH3), 11.2 (CH3), 13.8 (CH3), 15.4 (CH3), 20.1 (CH3), 20.8 (CH3), 21.0 (CH3), 21.2 (CH3), 22.7 (CH2), 25.6 (CH2), 26.0 (CH2), 26.5 (CH2), 47.1 (CH2), 49.9 (CH), 110.7 (ArCH), 120.0 (ArCH), 128.0 (ArCH), 129.6 (ArCH), 148.2 (C), 170.0 (C=O), 172.6 (C=O); IR (film): vmax(cm1) = 3312 (NH stretch), 2959, 2928, 2872, 1707, 1684, 1600, 1549, 1390, 1230; HRMS for the carboxylate-urea ion (C16H20N2O5) (ESI ): m / z measured = 319.1297 [M]’, predicted = 319.1289 [M]’.

[0333] SSA 141: (R)-2-Amino-3-(naphthalen-l-yl) propanoic acid (0.90 g, 4.20 mmol) was dissolved in methanol (10.00 mL) and tetrabutylammonium hydroxide in methanol (5.00 mL, 1.00 M) added. The solvent was removed under reduced pressure. The sample was further dried for 24 hours under high vacuum. The tetrabutylammonium salt was then dissolved in ethyl acetate (30.00 mL) and an inert atmosphere applied prior to adding 4-butylphenylisocyonate dropwise to the mixture. The mixture was then heated to 60 °C. After 48 hours the solvent was removed under reduced pressure and the sample was further dried for 24 hours under high vacuum giving a pale-yellow sticky precipitate as a pure product (2.20 g, 2.69 mmol, 87 %). Melting point: 351 K;XH NMR (400 MHz, 333 K, DMSO-d6): 6:Case Ref. P319WO IPTector®0.86-0.98 (m, 15H), 1.23-1.37 (m, 10H), 1.46-1.63 (m, 10H), 2.43-2.49 (m, 2H), 3.17 (t, J = 8.6 Hz, 8H), 3.25 - 3.32 (m, 1H), 3.47-3.54 (m, 1H), 4.07 (s, 1H), 6.97 (d, J = 8.4 Hz), 7.23 - 7.46 (m, 4H), 7.68 (d, J = 7.72 Hz, 2H), 7.82 - 7.86 (m, 1H), 7.78-7.92 (m, 1H), 8.38 (t, J = 3.8 Hz, 1H), 8.91 (s, 1H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 13.5 (CH3), 13.9 (CH3), 19.3 (CH2), 21.7 (CH2), 23.1 (CH2), 33.5 (CH2), 34.2 (CH2), 37.0 (CH2), 56.5 (CH), 57.5 (CH), 117.4 (ArCH), 124.8 (ArCH), 125.1 (ArCH), 125.3 (ArCH), 125.9 (ArCH), 127.5 (ArCH), 128.1 (ArCH), 132.7 (ArC), 133.30 (ArC), 133.6 (ArC), 136.6 (ArC), 139.3 (ArC), 155.4 (C=O), 172.9 (C=O); IR (film): vmax(cm1) = 3298 (NH stretch), 3042, 2957, 2929, 2872, 1686, 1603, 1541, 1379, 1233; HRMS for carboxylate ion (C24H25N2O3) (ESI ): m / z measured = 388.1798 [M]_, predicted = 388.1781 [M]\0 0TBA-O^ T N A N JU N N YII H H142H H H

[0334] Compound 142: 183 was dissolved in methanol (5 mL) and sonicated for 30 minutes with a 2.20 equivalence of tetrabutylammonium hydroxide (2.03 mL, 2.20 mmol) producing a carboxylate SSA. This mixture was taken to dryness in vacuum to give a white solid. This carboxylate SSA was recrystallised in ethyl acetate, with a yield of 87 %. Melting Point: 100 °C.XH NMR (400 MHz, 298.15 K, DMSO-dg): 6: 9.97 (s, 2H), 7.44 (d, J = 7.40 Hz, 4H), 6.96 (d, J = 6.96 Hz, 4H), 6.76 (t, J = 4.48 Hz, 2H), 3.72 (s, 4H), 3.36 (d, J = 2. 96 Hz, 4H), 3.16-3.12 (m, 16H), 1.53-1.49 (m, 16H), 1.33-1.24 (m, 16H), 0.91 (t, J = 7.28 Hz, 24H);13C{1H} NMR (100 MHz, 298.15 K, DMSO-d6): δ: 171.3 (C=O), 155.8 (C=O), 140.1 (ArC), 133.5 (ArC), 128.9 (ArCH), 117.7 (ArCH), 57.9 (CH2), 45.5 (CH2), 26.2 (CH2) 23.5 (CH2), 19.6 (CH2), 13.9 (CH3); IR (film) v = 3331 (NH stretch), 2960, 1682, 1597, 1302, 824; HRMS (ESI ) for the carboxylate ion (C19H18N4O62-[M-2H]2-: m / z: act: 398.1237 [M]’ cal: 398.1237 [M]’.

[0335] SSA 143: Glycine methyl ester hydrochloride (0.55 g, 2.00 mmol), 1, 4-phenylene diisocyanate (0.32 g, 2.00 mmol) and anhydrous triethylamine (0.79 mL, 4.40 mmol) was mixed overnight at RT in DCM to yield a white solid. After mixing, the solvent was evaporated under reduced pressure. The resultant solid was mixed in methanol (6 mL), deionised water (4 mL) and NaOH (1.80 mL, 4.00 mmol) for 4 hours at RT. After 4 hours, HCI at a 2 M concentration was added dropwise until the mixture became slightly acidic. The mixture was transferred straight to a separation funnel, where it was washed in DCM and water, yielding a solid white precipitate. The solid was filtered off and taken to dryness under reduced pressure. The dried solid was dissolved in 5 mL of methanol and sonicated for 30 minutes with tetrabutylammonium hydroxide added at a 2.20 equivalence (3.40 mL, 2.20 mmol). The product was taken to dryness under reduced pressure and was a dark orange solid. The finalCase Ref. P319WO IPTector®product sonicated in ethyl acetate for 30 minutes, which formed the final product as a light-yellow solid with a yield of 25 %. Melting Point: >205 °C.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 8.67 (s 2H), 7.19 (s, 4H), 5.99 (t, J = 4.08 Hz, 4H), 3.29 (d, J = 3.80 Hz, 4H), 3.18 (m, 16H), 1.59 (m, 16H), 1.49 (m, 16H), 0.94 (t, J = 7.24 Hz, 24H);13C{1H} NMR (100 MHz, 298.15 K, DMSO-d6): δ 170.9 (C=O), 155.6 (C=O), 135.1 (ArC), 118.3 (ArCH) ), 57.9 (CH2), 45.4 (CH2), 23.5 (CH2), 19.6 (CH2), 13.9 (CH3). IR (film) v = 3449 (NH stretch), 2963, 1717, 1593, 1300, 883; HRMS for carboxylate ion (C12H12N4O62-(ESI ): m / z: act: 308.0755 [M]’ cal: 308.0768.[M]'.

[0336] SSA 144: 4,4'-(Propane-2,2-diyl)dianiline (0.45 g, 2.00 mmol) and anhydrous triethylamine (0.79 mL, 4.40 mmol) was added to a stirring solution of triphosgene (0.59 g, 2 mmol) in ethyl acetate (50 mL) and refluxed at 80 °C under an inert atmosphere for 4 hours. Glycine (4 mmol, 0.30 g) was dissolved in tetrabutylammonium hydroxide in methanol (4 mL, 4 mmol) and evaporated under reduced pressure. The tetrabutylammonium glycine was then dissolved in acetonitrile (20 mL) and added to the stirring solution of isocyanate and refluxed at 80 °C under an inert atmosphere overnight. The resultant oil was sonicated for 30 minutes in ethyl acetate (20 mL). The ethyl acetate was decanted, and the remaining material was dried under reduced pressure to yield the product as a pink solid with a yield of 92 %. Melting Point: 138 °C.XH NMR (400 MHz, 298.15 K, DMSO-dg): 6: 9.07 (s, 2H), 7.28 (d, J = 8.52 Hz, 4H), 7.01 (d, J = 8.96 Hz, 4H),6.18 (t, J = 5.12 Hz, 2H), 3.23 (d, J = 3.60 Hz, 4H), 3.17 (m, 16H), 1.57 (m, 16H), 1.55 (s, 6H), 1.31 (m, 16H), 0.94 (t, J = 7.28 Hz, 24H);13C{1H} NMR (100 MHz, 298.15 K, DMSO-d6): δ 170.4 (C=O), 155.4 (C=O), 142.9 (ArC), 139.2 (ArC), 126.9 (ArCH), 117.4 (ArCH), 57.9 (CH2), 45.5 (C), 41.6 (CH2), 31.1 (CH3), 23.5 (CH2), 19.7 (CH2), 13.9 (CH3). IR (film) v = 3317 (NH stretch), 2960, 1685, 1597, 1300, 879; HRMS (ESI ) for the carboxylate ion (C21H22N4O62-[M-2H]2-: m / z: act: [M]’ cal: 213.0775 [M]2.

[0337] SSA 145: Glycine methyl ester hydrochloride (0.69 g, 5.50 mmol), 4-trifluoromethoxy benzene isocyanate (0.76 mL, 5.00 mmol) and anhydrous triethylamine (0.83 mL, 6.03 mmol) were stirred overnight at room temperature in dichloromethane (30 mL) to yield a white solid. The resultant precipitate (0.82 g, 2.81 mmol) was removed by filtration and redissolved in methanol (6.00 mL). Water (4.00 mL) and an aqueous solution of NaOH (3.08 mL, 2.00 M) were then added and left to stir for 3 hours at room temperature. HCI (2.00 M) was then added dropwise until pH = 5.00. The resultant precipitate (2.09 g, 7.51 mmol) was then filtered, washed with ethyl acetate (30 mL) and water (30 mL), re-dissolved in methanol (5.00 mL) and sonicated for 30 minutes with one equivalence of tetrabutylammonium hydroxide in methanol (7.51 mL, 1.00 M). The solution was then taken to dryness under reduced pressure to give a white solid. This white solid was then recrystallised in ethyl acetate (30 mL) and washed with water (30 mL) to give the pure product as a white solid (2.89 g, 5.56Case Ref. P319WO IPTector®mmol, 74 %). Melting point: 347 K;XH NMR (400 MHz, 298 K, DMSO-d6): δ: 0.91 (t, J = 7.2 Hz, 12H), 1.23-1.35 (m, 8H), 1.49-1.60 (m, 8H), 3.15 (t, J = 8.0 Hz, 8H), 3.43 (d, J = 2.3* Hz, 2H), 6.98 (s, 1H), 7.16 (d, J = 8.5 Hz, 2H), 7.64 (d, J = 8.8 Hz, 2H), 10.37 (s, 1H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 13.9 (CH3), 19.7 (CH2), 23.5 (CH2), 45.0 (CH2), 49.0 (CH2), 57.9 (s, ArC), 121.8 (q, J = 19.6 Hz, CF3), 141.6 (ArC), 141.7 (d, ArCH), 155.6 (C=O), 171.7 (C=O); IR (film): vmax(cm1) = 3331 (NH stretch) 1695, 1487, 1257, 1153, 877; HRMS for the carboxylate-urea ion (C10H8F3N2O4) (ESI ): m / z measured = 277.0428 [M]’, predicted = 277.0442 [M]’. *J-coupling value is difficult to calculate due to overlap with water peak.

[0338] SSA 146: D-Glutamic acid dimethyl ester hydrochloride (1.16 g, 5.50 mmol), 4-trifluoromethoxy benzene isocyanate (0.76 mL, 5.00 mmol) and anhydrous triethylamine (0.83 mL, 6.03 mmol) were stirred overnight at room temperature in dichloromethane (30 mL) to yield a pinkishgrey waxy solid. The resultant precipitate (1.54 g, 4.07 mmol) was removed by filtration and redissolved in methanol (6.00 mL). Water (4.00 mL) and an aqueous solution of NaOH (4.46 mL, 2.00 M) were then added and left to stir for 3 hours at room temperature. HCI (2.00 M) was then added dropwise until pH = 5.00. The resultant precipitate (0.80 g, 2.28 mmol) was then filtered, washed with ethyl acetate (30 mL) and water (30 mL), re-dissolved in methanol (5.00 mL) and sonicated for 30 minutes with two equivalences of tetrabutylammonium hydroxide in methanol (4.56 mL, 1.00 M). The solution was then taken to dryness under reduced pressure to give a brownish-white sticky solid (1.37 g, 1.66 mmol, 60 %). Melting point: 350 K;TH NMR (400 MHz, 343 K, DMSO-d6): 6: 0.95 (br s, 12H), 1.34 (br s, 8H), 1.61 (br s, 8H), 1.77 (s, 1H), 1.99 (s, 1H), 2.15 (br s, 1H), 3.20 (br s, 8H), 3.76 (s, 1H), 6.28 (s, 1H), 7.05 (s, 2H), 7.76 (s, 2H), 10.98 (s, 1H);13C{1H} NMR (100 MHz, 343 K, DMSO-d6): δ: 13.7 (CH3), 19.7 (CH2), 23.7 (CH2), 35.6 (CH2), 49.0 (CH), 55.8 (CH2), 58.5 (CH2), 119.5 (q, J = 104 Hz, CF3), 142.0 (ArCH), 142.2 (ArC), 156.1 (ArC), 174.2 (C=O), 176.9 (C=O); IR (film): vmax(cm1) = 3334 (NH stretch), 2962, 1697, 1253, 1047, 844; HRMS for the carboxylate-urea ion (C13H11F3N2O5) (ESI ): m / z measured = 348.0596 [M]’, predicted = 348.0580 [M]’.

[0339] SSA 147: (R)-3-(l-Naphthyl)-D-alanine (1.18 g, 5.50 mmol) was dissolved in methanol (5.00 mL) and sonicated for 10 minutes with one equivalence of tetrabutylammonium hydroxide in methanol (5.50 mL, IM) producing a naphthyl TBA salt which was dried under pressure overnight. This brown-oil was then added to a solution of 4-trifluoromethoxy benzene isocyanate (0.76 mL, 5.00 mmol) and anhydrous triethylamine (0.83 mL, 6.03 mmol), then stirred overnight at room temperature in dichloromethane (30 mL) to yield an orange-brown solid. The resultant mixture was taken to dryness under reduced pressure. The crude product was purified using silica flash chromatography (100 % ethyl acetate followed by 100 % methanol). The resulting brownish-gold glassy product was dried under reduced pressure overnight (3.00 g, 4.30 mmol, 86 %). Melting point:Case Ref. P319WO IPTector®347 K; NMR (400 MHz, 343 K, DMSO-d6): δ: 0.91 (t, J = 7.2 Hz, 12H), 1.23-1.34 (m, 8H), 1.48-1.59 (m, 8H), 3.13 (t, J = 8.4 Hz, 8H), 3.34-3.41 (m, 2H), 4.31 (s, 1H), 7.10-8.02 (m, 11H), 8.42 (d, J = 8.0 Hz, 1H), 10.24 (s, 1H);13C{1H} NMR (100 MHz, 343 K, DMSO-d6): δ: 13.5 (CH3), 19.2 (CH2), 23.1 (CH2), 56.0 (CH), 57.5 (CH2), 118.3 (q, J = 70 Hz, CF3), 124.4 (ArCH), 125.2 (ArCH), 125.3 (ArCH), 125.5 (ArCH), 127.5 (ArC), 128.3 (ArC), 132.3 (ArC), 133.3 (ArC), 135.8 (ArC), 140.9 (ArC), 141.3 (d, J = 1.8 Hz, ArCH), 155.2 (C=O), 173.9 (C=O); IR (film): vmax(cm1) = 3330 (NH-stretch), 2962, 1685, 1193, 1016, 777; HRMS for the carboxylate-urea ion (C21H19N3O3) (ESI j: m / z measured = 417.1065 [M]’, predicted = 417.1068 [M]’

[0340] SSA 158: (R)-3-(l-Naphthyl)-D-alanine (0.86 g, 4.00 mmol) and tetrabutylammonium hydroxide (4.0 mL, 4.00 mmol) were sonicated for 30 minutes, until all solid was dissolved, before taking to complete dryness under reduced pressure. The resultant solid was dissolved in acetonitrile (20 mL) and added to a stirring solution of ethyl acetate (20 mL) and 4,4'-Methylenebis phenyl isocyanate (0.50 g, 2.00 mmol) and refluxed at 80 °C overnight. After 12 hours, the reaction yielded a black oil. The supernatant solvent was decanted to isolate the oil, which was subsequently sonicated in ethyl acetate and dried under reduced pressure to afford the TBA salt as a brown solid with a yield of 84 %.1H NMR (400 MHz, 298.15 K, DMSO-dg): 6: 9.03 (d, 2H, J = 3.36 Hz), 3.38 (t, 2H, J = 4 Hz), 7.68 (d, 2H, J = 7.8 Hz), 7.42 (m, 4H), 7.34 (m, 8H), 6.98 (d, 4H, J = 8.52 Hz), 6.45 (s, 2H), 4.05 (q, 2H, J = 5.56 Hz), 3.72 (s, 2H), 3.49 (dd, 2H, J = 6.20 Hz), 3.29 (dd, 2H, J = 5.12 Hz), 3.15 (m, 16H), 1.55 (m, 16H), 1.30 (m, 16H), 0.93 (t, 24H, J = 7.28 Hz);13C{1H} NMR (100 MHz, 298.15 K, DMSO-d6): δ: 173.5 (C=O), 155.8 (C=O), 140.1 (ArC), 137.0 (ArC), 133.7 (ArC), 133.5 (ArC), 133.1 (ArC), 129.2 (ArCH), 128.9 (ArCH), 128.6 (ArCH), 127.9 (ArCH), 126.3 (ArCH), 125.8 (ArCH), 125.7 (ArCH), 125.5 (ArCH), 125.2 (ArCH), 117.9 (ArCH), 57.9 (CH), 57.0 (CH), 40.4 (CH), 37.6 (CH2), 23.5 (CH2), 19.6 (CH2), 13.9 (CH3). IR (film) v = 3323 (NH stretch), 2960, 1685, 1541, 1307, 777.

[0341] SSA 161: The (aminomethyl)phosphonic acid (0.37 g, 3.30 mmol) in methanol (10 mL) was combined with IN tetrabutylammonium hydroxide in methanol (6.60 mL, 6.60 mmol) before being taken to dryness. The 4-(trifluoromethyl)phenyl isocyanate (0.43 mL, 3.00 mmol) was combined with (aminomethyl)phosphonate tetrabutylammonium in ethyl acetate (30 mL) and heated at reflux underCase Ref. P319WO IPTector®nitrogen for 48 hours. The resulting reaction mixture was taken to dryness. The solid was redissolved in dichloromethane (30 mL) and washed with water (2x 30 mL). The aqueous layer was then concentrated under reduced pressure until a precipitate formed. The precipitate was filtered under reduced pressure and the pure product was obtained, a white solid with a yield of 20.4 % (0.33 g, 0.60 mmol); melting point >200 °C;TH NMR (400 MHz, 298 K, DMSO-d6): 6 0.91 (t, J = 8.0 Hz, 12H), 1.25- 1.34 (sext, J = 8.0 Hz, 8H), 1.51-1.59 (m, J = 8.0 Hz, 8H), 3.03-3.07 (m, 2H), 3.13-3.17 (m, J = 8.0 Hz, 8H), 7.38-7.40 (m, J = 8.0 Hz, 3H), 7.59-7.61 (d, J = 8.0 Hz, 2H), 10.69 (s, 1H);13C{1H} NMR (100 MHz, 298 K, DMSO-dg): δ 13.4 (CH3), 19.2 (CH2), 23.1 (CH2), 38.6 (d, J = 140 Hz, CH2), 57.5 (CH2), 116.7 (ArCH), 119.1 (q, J = 30.0 Hz, ArC), 120.8 (q, J = 270.0 Hz, CF3), 125.3-125.4 (m, J = 3.0 Hz, ArCH), 145.5 (ArC), 155.7 (d, J = 10.0 Hz, CO); IR (film): v = 3491 cm1(O-H Stretch), 3315 cm1(N-H Stretch), 2961 cm1, 2936 cm' 2876 cm1(C-H stretch), 1701 cm1(C=O stretch), 1605 cm1, 1553 cm'^C-C stretch), 1321 cm1, 1240 cm-1(C-0 stretch), 1177 cm1, 1153 cm'1(P=O stretch), 1115 cm1, 1088 cm1, 1062 cm'1(P-0 stretch), 910 cm1, 853 cm1(C-H bend); HRMS for the hydrogen phosphonate-urea ion (C9H9F3N2O4P-)(ESI-): m / z: act 297.0257 [M-H]' cal: 297.0239 [M-H]'.

[0342] SSA 162: The (aminomethyl)phosphonic acid (0.74 g, 6.60 mmol) in methanol (10 mL) ) was combined with IN tetrabutylammonium hydroxide in methanol (13.20 mL, 13.30 mmol) before being taken to dryness. The 4-(trifluoromethyl)phenyl isothiocyanate (1.22 mL, 6.00 mmol) in ethyl acetate (40 mL) was combined with (aminomethyl)phosphonate tetrabutylammonium and stirred at room temperature for 96 hours. The resulting mixture was taken to dryness and then redissolved with ethyl acetate (30 mL). The organic layer was washed with water (2x 30 mL) which formed a precipitate. The precipitate was filtered under reduced pressure to obtained the pure product, a white solid with a yield of 66.7 % (2.2 g, 4.00 mmol); melting point >200 °C;1H NMR (400 MHz, 298 K, DMSO-d6): δ 0.91 (t, J = 8.0 Hz, 12H), 1.25 (sext, J = 8.0 Hz, 8H), 1.51-1.59 (m, J = 8.0 Hz, 8H), 3.13-3.17 (m, J = 8.0 Hz, 8H), 3.34 (dd, J = 16.0 Hz, 2H), 7.52 (d, J = 12.0 Hz, 2H), 8.0 (d, J = 8.0 Hz, 2H), 9.18-9.20 (m, J = 8.0 Hz, 1H), 11.85 (s, 1H);13C{1H} NMR (100 MHz, 353 K, DMSO-d6): δ 12.8 (CH3), 18.8 (CH2), 22.9 (CH2), 43.5 (d, J = 130 Hz, CH2), 57.7 (CH2), 120.1 (bs, ArCH), 120.2 (q, J = 270.0 Hz, CF3), 121.1 (q, J = 30.0 Hz, ArC), 124.3 (m, J = 3.0 Hz, ArCH), 144.9 (ArC), 180.8 (d, J = 10.0 Hz, CS); IR (film): v = 3233 cm'1(N-H Stretch), 2954 cm1, 2874 cm1, 2733 cm'1(O-H stretch), 1518 cm'^C-C stretch), 1319 cm'1(C-0 stretch), 1152 cm'1(P=O stretch), 1117 cm1, 1092 cm1, 1065 cm'1(P-0 stretch), 925 cm1, 880 cm1(C-H bend); HRMS for the hydrogen phosphonate-urea ion (C9H9F3N2O3PS (ESI: m / z: act 397.0257 [M-H]' m / z: act 313.0062 [M-H]' cal: 313.0015 [M-H]'.Case Ref. P319WO IPTector®

[0343] SSA 165: (R)-2-amino-3-(napthalen-2-yl)propanoic acid (0.90 g, 4.20 mmol) and tetrabutylammonium hydroxide in methanol (4.20 mL, IM) was dissolved in methanol (10.00 mL), heated, sonicated, and dried under reduced pressure.4-(6-Methyl-l,3-benzothiazol-2-yl)phenylamine (1.92 g, 8.00 mmol) was dissolved in ethyl acetate (30.00 mL) and together with triethylamine (1.00 mL, 7.20 mmol) added to the mixture. Triphosgene (1.18 g, 4.00 mmol) was added to the solution and refluxed for 4 hours. (R)-2-Amino-3-(naphthalen-2-yl)propanoate tetrabutylammonium salt was added to the reaction with acetonitrile (10.00 mL) and reacted overnight. The resulting crude yielded a sticky brown precipitate, the solvent was decanted and taken to dryness. Redissolving in dichloromethane (30.00 mL) and extracting with deionised water (30.00 mL x2), the crude formed a precipitate. The precipitate was resuspended in methanol and refiltered under reduced pressure. Hydantoin hydrolysis was performed with methanol: isopropyl alcohol (5.00 mL: 5.00 mL) and heating to 50 ° C. Sodium hydroxide (4.00 mL, 4.00 mmol) was added and reacted for 16 hours at room temperature. Subsequently, a pH = 1.00 was attained with hydrochloric acid (2M) before flooding with deionised water and filtering under reduced pressure. The pure product was a light brown solid (1.40 g, 1.9 mmol, 24 %). Melting point: > 473 K;XH NMR (400 MHz, 298 K, DMSO-d6): δ 0.89 (t, J = 7.4 Hz, 12H), 1.23-1.32 (m, 8H), 1.47-1.55 (m, 8H), 2.45 (s, 3H), 3.10 (t, J = 8.4 Hz, 8H), 3.45-3.55 (m, 2H), 4.24-4.27 (q, J = 7.6 Hz, 1H), 7.30-7.53 (m, 6H), 7.72-7.88 (m, 8H), 8.50 (d, J = 8.1 Hz, 1H), 10.72 (s, 1H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 13.5 (CH3), 19.2 (CH2), 21.0 (CH3), 23.0 (CH2), 37.1 (CH2), 56.53 (CH), 57.5 (CH2), 117.3 (ArCH), 121.6 (ArCH), 121.8 (ArCH), 124.5 (ArC), 124.7 (ArCH), 125.1 (ArCH), 125.3 (ArCH), 126.0 (ArCH), 127.4 (ArCH), 127.6 (ArCH), 127.8 (ArCH), 128.2 (ArCH), 132.6 (ArC), 133.3 (ArC), 134.5 (ArC), 136.3* (ArC), 144.9 (ArC), 151.9 (ArC), 155.0 (CO), 166.5 (ArC) 173.1 & 173.2* (CO); IR (film): vmax(cm1) = 3298 (NH Stretch), 3041, 2958, 2872, 1695, 1595, 1479, 1452, 1379, 1319, 1225, 1175; HRMS for the carboxylate-urea ion 10 (C28H22N3O3S)(ESI j: m / z measured = 480.1414 [M]’, predicted = 480.1419 [M]’. *Conformational isomerism observed in this molecule.169Case Ref. P319WO IPTector®

[0344] SSA 169: (R)-3-(l-Naphthyl)-D-alanine (1.18 g, 5.50 mmol) was dissolved in methanol (5.00 mL) and sonicated for 10 minutes with one equivalent of tetrabutylammonium hydroxide in methanol (5.50 mL, IM) producing a naphthyl-tetrabutylammonium salt which was then dried under reduced pressure overnight. This brownish-gold oil was then added to a solution of 4-cyanophenyl isocyanate (0.721 g, 5.00 mmol) and anhydrous triethylamine (0.83 mL, 6.03 mmol), then stirred overnight at room temperature in dichloromethane to yield an orange-brown solid. The resultant precipitate was taken to dryness under reduced pressure. The crude product was purified using silica flash chromatography (ethyl acetate followed by methanol. The resulting golden glassy product was dried under reduced pressure overnight (2.57 g, 4.28 mmol, 78 %). Melting Point: 350 K;1H NMR (400 MHz, 298 K, DMSO-dg): 6: 0.91 (t, J = 7.2 Hz, 12H), 1.23-1.33 (m, 8H), 1.48-1.58 (m, 8H), 3.13 (t, J = 8.1 Hz, 8H), 3.41 (d, J = 4.8 Hz, 2H), 4.28 (s, 1H), 7.29-7.89 (m, 11H), 8.45 (d, J = 7.9 Hz, 1H), 10.96 (s, 1H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 13.5 (CH3), 19.2 (CH2), 23.0 (CH2), 56.4 (CH), 57.5 (CH2), 101.0 (ArCH), 117.3 (ArCH), 119.8 (CH), 124.4 (ArCH), 125.33 (t, J = 16.2 Hz, CH2), 126.2 (ArCH), 127.4 (ArC), 128.3 (ArC), 132.4 (ArC), 132.8 (ArC), 133.3 (ArC), 135.8 (ArC), 146.2 (CN), 154.8 (C=O), 173.7 (C=O); IR (film): vmax (cm1) = 3284 (NH stretch), 2214, 1593, 1171, 842; HRMS for the carboxylate-urea ion 7 (C21H16N3O3) (ESI ): m / z measured = 358.1186 [M]’, predicted = 358.1187 [M]’.

[0345] SSA 170: (R)-3-(l-Naphthyl)-D-alanine (1.18 g, 5.50 mmol) product was dissolved in methanol (5.00 mL) and sonicated for 10 minutes with one equivalence of tetrabutylammonium hydroxide (5.50 mL, 5.50 mmol) producing a naphthyl-tetrabutylammonium salt which was dried under reduced pressure overnight. This brownish-gold salt-oil was then added to a solution of 5-cyanophenyl isocyanate (0.72 g, 5.00 mmol) and anhydrous triethylamine (0.83 mL, 6.03 mmol), then stirred overnight at room temperature in dichloromethane to yield a dark orange-brown solid. The resultant precipitate was taken to dryness under reduced pressure. The crude product was purified using silica flash chromatography (ethyl acetate followed by methanol. The resulting brown glassy product was dried under reduced pressure overnight (2.15 g, 3.58 mmol, 65 %). Melting Point: 350 K;XH NMR (400 MHz, 298 K, DMSO-dg): 6: 0.91 (t, J = 7.6 Hz, 12H), 1.23-1.34 (m, 8H), 1.49-1.59 (m, 8H), 3.09-3.17 (m, 8H), 3.41 (t, J = 7.6 Hz, 2H), 4.31 (s, 1H), 7.13-8.13 (m, 11H), 8.46 (d, J = 8.2 Hz, 1H), 10.80 (s, 1H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 13.5 (CH3), 19.2 (CH2), 23.0 (CH2), 56.4 (CH), 57.5 (CH2), 111.1 (ArC), 119.2 (ArC), 119.6 (CH), 122.0 (ArCH), 123.3 (ArCH), 124.4 (ArCH), 125.30 (t, J = 19.8 Hz,Case Ref. P319WO IPTector®CH2), 126.2 (ArCH), 127.4 (ArCH), 128.3 (ArCH), 129.6 (ArC), 132.3 (ArC), 133.3 (ArC), 135.8 (ArC), 142.6 (CN), 155.1 (C=O), 174.0 (C=O); IR (film): vmax(cm-1) = 3275 (NH stretch), 2224, 1585, 1232, 777; HRMS for the carboxylate-urea ion 8 (C21H16N3O3) (ESI ): m / z measured = 358.1187 [M]’, predicted = 358.1197 [M]-.

[0346] SSA 171: (R)-3-(l-Naphthyl)-D-alanine (1.07 g, 5.00 mmol) was dissolved in methanol (5.00 mL) and sonicated for 10 minutes with one equivalent of tetrabutylammonium hydroxide in methanol (5.50 mL, 1.00 M) producing a naphthyl tetrabutylammonium salt which was dried under reduced pressure overnight. In a separate round bottom, triphosgene (0.445 g, 1.5 mmol) was added to a stirring solution of 4-aminophenylsulphur pentafluoride (1.10 g, 5.00 mM), anhydrous triethylamine (1.25 mL, 12.32 mmol) and dichloromethane (30 mL), and the mixture heated at reflux for 4 hours. The naphthyl tetrabutylammonium salt was then added to the solution and anhydrous triethylamine (0.83 mL, 6.03 mmol) was topped up, then stirred overnight at room temperature to yield a cream white solid. The resultant precipitate was taken to dryness under reduced pressure. The crude product was purified using silica flash chromatography (ethyl acetate followed by methanol). The resulting cream white product was dried under vacuum overnight (1.75 g, 2.49 mmol, 50 %). Melting Point: 355 K;1H NMR (400 MHz, 298 K, DMSO-d6): δ: 0.91 (t, J = 7.3 Hz, 12H), 1.23-1.34 (m, 8H), 1.48-1.58 (m, 8H), 3.13 (t, J = 7.9 Hz, 8H), 3.35 (s, 2H), 4.19 (s, 1H), 7.29-7.89 (m, 11H), 8.44 (d, J = 7.9 Hz, 1H), 10.94 (s, 1H);13C{1H} NMR (100 MHz, 298 K, DMSO-d6): δ: 13.4 (CH3), 19.2 (CH2), 23.0 (CH2), 56.5 (CH), 57.5 (CH2), 116.3 (CH), 124.6-126.2 (m, C-SF5), 127.4 (ArCH), 128.2 (ArCH), 132.5 (ArCH), 133.3 (ArCH), 136.1 (ArCH), 144.3-144.6 (m, C-SF5), 145.2 (C=O), 154.8 (C=O); IR (film): vmax(cm-1) = 3290 (NH stretch), 2960, 1485, 1097, 833, HRMS for the carboxylate-urea ion 9 (C20H16F5N2O3S) (ESI ): m / z measured = 459.0801 [M]’, predicted = 459.0807 [M]’.F3C CF3V N N N N Y TBA J H H 172 H H " TBA

[0347] SSA 172: 185 was dissolved in methanol (5 mL) and sonicated for 30 minutes with a 2.20 equivalence of tetrabutylammonium hydroxide (1.44 mL, 1.44 mmol) and taken to dryness under reduced pressure. The resultant oil was washed in ethyl acetate, and the organic solution was decanted, and the remaining oil was dried under reduced pressure, yielding a pure-white solid with aCase Ref. P319WO IPTector®yield of 53 %. Melting Point: 164 °C.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 9.41 (s, 2H), 7.46 (d, J = 8.64 Hz, 4H), 7.14 (d, J = 8.48 Hz, 4H), 6.31 (t, J = 8.04 Hz, 2H), 3.25 (d, J = 3.00 Hz, 4H), 3.16 (m, 16H), 1.57 (m, 16H), 1.31 (m, 16H), 0.94 (t, J = 7.24 Hz, 24H);13CfH} NMR (100 MHz, 298.15 K, DMSO-d6): δ: 170.9 (C=O), 155.5 (C=O), 143.2 (ArC), 130.2 (ArC), 129.1 (q, J = 282.5 Hz CF3), 123.5 (ArCH), 117.0 (ArCH), 63.8 (q, J = 22.3, C), 58.0 (CH2), 45.5 (CH2), 23.5 (CH2), 19.6 (CH2), 13.9 (CH2). IR (film) v = 3323 (NH Stretch), 2873, 1685, 1600, 1307, 879; HRMS for the carboxylate ion (C2iHi6F6N4O62(ESI ): m / z: act: 534.0985 [M] cal: 534.0944 [M]’.173

[0348] SSA 173: Compound 183 (0.20 g, 0.50 mmol) and chlorhexidine (0.25 g, 0.50 mmol) were dissolved in methanol and sonicated at 45 °C for 2 hours. The organic layer was evaporated under reduced pressure to yield a white crystalline solid.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 11.10 (s, 2H), 8.81 (s, 2H), 7.28 (m, 20H), 6.11 (s, 2H), 3.74 (s, 2H), 3.49 (s, 4H), 3.07 (s, 4H), 1.35 (m, 8H).174

[0349] SSA 174: Compound 184 (0.21 g, 0.50 mmol) and chlorhexidine (0.25 g, 0.50 mmol) were dissolved in methanol and sonicated at 45 °C for 2 hours. The organic layer was evaporated under reduced pressure to yield a pink crystalline solid.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 11.19 (s, 2H), 8.83 (s, 2H), 7.33 (m, 20H), 6.16 (t, J = 3.80 Hz), 3.53 (s, 4H), 3.12 (t, J = 6.16 Hz, 4H), 1.62 (s, 6H), 1.40 (m, 8H).175

[0350] SSA 175: Compound 185 (0.27 g, 0.50 mmol) and chlorhexidine (0.25 g, 0.50 mmol) were dissolved in methanol and sonicated at 45 °C for 2 hours. The organic layer was evaporated under reduced pressure to yield a yellow crystalline solid.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 11.13 (s, 2H), 9.22 (s, 2H), 7.28 (m, 20H), 6.31 (t, J = 4.16 Hz, 2H), 3.50 (s, 4H), 3.07 (s, 4H), 1.36 (m, 8H).176

[0351] SSA 176: Compound 183 (0.20 g, 0.50 mmol) and octenidine (0.28 g, 0.50 mmol) were dissolved in ethanol and sonicated at 45 °C for 2 hours. The organic layer was evaporated underCase Ref. P319WO IPTector®reduced pressure to yield a white crystalline solid.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 12.53 (s, 2H), 8.70 (m, 4H), 8.29 (d, J = 7.36 Hz, 2H), 8.12 (d, J =7.68 Hz, 2H), 7.28 (d, J = 8.52 Hz, 4H), 7.05 (d, J = 8.40 Hz, 4H), 6.91 (m, 4H), 6.32 (t, J = 6.24 Hz, 2H), 4.09 (t, J = 7.32 Hz, 4H), 3.76 (s + d, J = 6.08 Hz, 6H), 3.25 (q, J = 6.20 Hz, 4H), 1.73 (m, 4H), 1.56 (m, 4H), 1.26 (m, 32H), 0.86 (m, 6H).

[0352] SSA 177: Compound 184 (0.21 g, 0.50 mmol) and octenidine (0.28 g, 0.50 mmol) were dissolved in ethanol and sonicated at 45 °C for 2 hours. The organic layer was evaporated under reduced pressure to yield a pink crystalline solid.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 12.54 (s, 2H), 8.91 (t, J = 5.92 Hz, 2H), 8.22 (s, 2H), 8.30 (m, 2H), 8.15 (m, 2H), 7.27 (d, J = 8.68 Hz, 4H), 7.27 (d, J = 8.80 Hz, 4H), 6.93 (m, 4H), 6.40 (t, J = 5.72 Hz), 4.10 (t, 6.72 Hz, 4H), 3.77 (d, J = 6.08 Hz, 4H), 3.24 (q, J = 6.28 Hz, 4H), 1.72 (m, 4H), 1.56 (m, 10H), 1.26 (m, 32H), 0.86 (m, 6H).

[0353] SSA 178: Compound 185 (0.27 g, 0.50 mmol) and octenidine (0.28 g, 0.50 mmol) were dissolved in ethanol and sonicated at 45 °C for 2 hours. The organic layer was evaporated under reduced pressure to yield a yellow crystalline solid.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 12.59 (s, 2H), 9.27 (s, 2H), 8.29 (t, J = 5.52 Hz, 2H), 8.30 (m, 2H), 8.13 (m, 2H), 7.48 (d, J = 9.08 Hz, 4H), 7.17 (d, J = 9.00 Hz, 4H), 6.93 (m, 4H), 6.58 (t, J = 5.64 Hz, 2H), 4.10 (t, J = 7.48 Hz, 4H), 3.80 (d, J = 6.32 Hz, 4H), 3,24 (q, J = 6.84 Hz, 4H), 1.72 (m, 4H), 1.56 (m, 4H), 1.25 (m, 32H), 0.86 (m, 6H).

[0354] SSA 181: Compound 186 (0.34 g, 0.50 mmol) and chlorhexidine (0.25 g, 0.50 mmol) were dissolved in methanol and sonicated for at 45 °C for 2 hours. The organic layer was evaporated under reduced pressure to yield a light-brown solid.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 11.17 (s, 2H), 8.63 (s, 2H), 8.33 (t, 2H, J = 3.44 Hz), 7.87 (t, 2H, J = 5.27 Hz), 7.73 (d, 2H, J = 6.92 Hz), 7.36 (m, 26H), 7.00 (d, 4H, J = 8.48 Hz), 6.22 (d, 2H, J = 7.04 Hz), 4.31 (q, 2H, J = 6.28 Hz), 3.61 (m, 4H), 3.26 (q, 2H, J = 6.36 Hz), 3.05 (s, 4H), 1.42 (s, 4H), 1.24(s, 4H).Case Ref. P319WO IPTector®

[0355] SSA 182: Compound 186 (0.34 g, 0.50 mmol) and octenidine (0.28 g, 0.50 mmol) were dissolved in ethanol and sonicated at 45 °C for 2 hours. The organic layer was evaporated under reduced pressure to yield a brown crystalline solid.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 12.85 (s, 2H), 8.79 (s, 2H), 8.63 (s, 2H), 8.29 (d, 2H, J = 6.52 Hz), 8.18 (dd, 4H), 7.94 (d, 2H, J = 7.64 Hz), 7.81 (d, 2H, J = 7.76 Hz), 7.54 (m, 4H), 7.41 (m, 4H), 7.23 (d, 4H, J = 7.96 Hz), 6.92 (d, 4H, J = 6.48 Hz), 6.52 (d, 2H, J = 7.56 Hz), 4.55 (q, 2H, J = 5.32 Hz), 4.10 (t, 4H, J = 6.16 Hz), 3.73 (s, 2H), 3.59 (q, 2H, 5.28 Hz), 3.27 (q, 4H, J = 5.88 Hz), 1.76 (m, 4H), 1.59 (m, 4H), 1.26 (m, 32H), 0.87 (m, 6H).

[0356] SSA 183: Glycine methyl ester hydrochloride (0.55 g, 2.00 mmol), 4,4'-Methylenebis phenyl isocyanate (0.50 g, 2.00 mmol) and anhydrous triethylamine (0.79 mL, 4.40 mmol) was mixed overnight at RT in DCM to yield a white solid. The resultant precipitate was taken to dryness under reduced pressure. The product was mixed in methanol (6 mL), deionised water (4 mL) and NaOH (0.79 mL, 4.00 mmol) for 4 hours at RT. After mixing, HCL (2 M) was added dropwise until the mixture became slightly acidic. The resultant precipitate was filtered and washed with methanol and water, to give a white solid, with a yield of 89%. Melting Point: 198 °C.1H NMR (400 MHz, 298.15 K, DMSO-d6): δ: 12.50 (s, 2H), 8.69 (s, 2H), 7.28 (d, J = 8.56 Hz, 4H), 7.04 (d, J = 8.52 Hz, 4H), 6.31 (t, J = 5.80 Hz, 2H), 3.77 (d, J = 5.80 Hz, 4H), 3.75 (s, 2H);13C{1H} NMR (100 MHz, 298.15 K, DMSO-d6): δ: 172.7 (C=O), 155.7 (C=O), 138.6 (ArC), 134.9 (ArC), 129.3 (ArCH), 118.3 (ArCH), 41.8 (CH2), 40.2 (CH2). IR (film) v = 3352 (NH stretch), 1716, 1631, 1591, 1300, 852.

[0357] SSA 184: 4,4'-(Propane-2,2-diyl)dianiline (0.45 g, 2.00 mmol), ethyl isocyanatoacetate (0.26 g, 2.00 mmol) and anhydrous triethylamine (0.79 mL, 4.40 mmol), were refluxed overnight at 75 °C in ethyl acetate (30 mL) and acetonitrile (20 mL) under an inert atmosphere. The organic solution was taken to dryness under reduced pressure, and the resultant pink solid was dissolved in methanol (5 mL) and reprecipitated with water (40 mL). The pink solid was filtered under reduced pressure. The product was mixed in methanol (6 mL), deionised water (4 mL) and NaOH (0.79 mL, 4.00 mmol) for 4 hours at RT. After mixing, HCL (2 M) was added dropwise until the mixture became slightly acidic. The resultant precipitate was filtered and washed with methanol and water, to give a pink solid, with a yield of 84%. Melting Point: 176 °C.XH NMR (400 MHz, 298.15 K, DMSO-dg): 6: 12.57 (s, 2H), 8.69 (s,Case Ref. P319WO IPTector®2H), 7.26 (d, J = 8.68 Hz, 4H), 7.05 (d, J = 8.64 Hz, 4H), 6.30, (t, J = 5.72 Hz, 2H), 3.77 (d, J = 5.72 Hz, 4H), 1.56 (s, 6H);13C{1H} NMR (100 MHz, 298.15 K, DMSO-d6): δ: 172.7 (C=O), 155.7 (C=O), 143.9 (ArC), 138.2 (ArC), 127.1 (ArCH), 117.9 (ArCH), 41.8 (C), 41.8 (CH2), 31.0 (CH3). IR (film) v = 3313 (NH stretch), 2966, 1718, 1600, 1315, 829. 184: HRMS for carboxylate ion (C21H22N4O62-) (ESI2-): m / z: act: 428.1695 [M]-cal: 428.1695[M]-.

[0358] SSA 185: 2,2-Bis (4-aminophenyl)hexafluoropropane (0.69 g, 2 mmol), anhydrous triethylamine (0.79 mL, 4.40 mmol) and ethyl isocyanatoacetate (0.26 g, 2 mmol) were added to acetonitrile (30 mL) and refluxed at 82 °C under an inert atmosphere overnight. The resultant mixture was taken to dryness under reduced pressure and re-precipitated in methanol (10 mL) and water (30 mL). The white solid was dried under reduced pressure and added to a stirring solution of methanol (6 mL), deionised water (4 mL) and NaOH (0.79 mL, 4.00 mmol) for 4 hours at RT. After mixing, HCL (2 M) was added dropwise until the mixture became slightly acidic. The resultant precipitate w...

Claims

Case Ref. P319WO IPTector®Claims1. A compound of Formula (A2):FR2yX N 1 NA / nY7AR1H HFormula (A2);wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selected from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;n is any integer, such as from 1 to 6;X is selected from the group consisting of: S, O, NH, NH2+, NR10, NHR10+, and SR10+; and wherein each Ri, R2, R3, R4, R5, R6, R7, and R10is independently selected from the group consisting of: hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, such as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino, nitro, cyano, carboxyl, sulfonyl, phosphonyl, or fused with another one of R1, R2, R3, R4, R5, R6, R7, or R10of another compound of Formula (A2) forming a dimer between two compounds of Formula (A2); and wherein a pair of R1and R2, R2and R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of R1and R2, R2and R3, R3and R4, or R4and R5is bound in formula A2.

2. The compound of claim 1, wherein the compound is capable of self-associating in a liquid medium.Case Ref. P319WO IPTector®3. The compound of any one of the preceding claims, wherein the compound is an amphiphile.

4. The compound of any one of the preceding claims, wherein the compound is a self-associating amphiphile (SSA).

5. The compound of any one of the preceding claims, wherein the compound in a polar organic solvent, such as DMSO-dg 0.5 % H2O, exhibits a Kdimof from 15 to 140 M1, such as from 45 to 135 M1.

6. The compound of any one of the preceding claims, wherein the compound in a polar organic solvent, such as DMSO-d6(optionally with trace water), exhibits a dHof from 0.8 to 2.0 nm, such as from 1.4 to 1.9 nm.

7. The compound of any one of the preceding claims, wherein the compound exhibits an electrostatic surface potential energy maximum (Emax) of from -20 to -60 kJ. mol1, such as from -46 to -50 kJ·mol-1.

8. The compound of any one of the preceding claims, wherein the compound exhibits an electrostatic surface potential energy minimum (Emin) of from -650 to -800 kJ. mol1, such as from -700 to -725 kJ·mol-1.

9. The compound of any one of the preceding claims, wherein the compound exhibits an electrostatic surface potential energy minimum (Emin) as calculated by Spartan '24 of from 0 kJ-mol-1to -250 kJ-mol-1.

10. The compound of any one of the preceding claims, wherein the compound exhibits an electrostatic surface potential energy maximum (Emax) as calculated by Spartan '24 of from - 650 kJ-mol-1to -1050 kJ-mol-1.

11. The compound of any one of the preceding claims, wherein the compound has a cLogP value as calculated by SwissADME of less than 4.

12. The compound of any one of the preceding claims, wherein the compound in a polar organic solvent exhibits a dimerization constant (Kdim) of less than 3750 M-1.Case Ref. P319WO IPTector®13. The compound of any one of the preceding claims, wherein the compound has a logP of from 2.50 to 5.50, for example from 3.0 to 4.50.

14. The compound of any one of the preceding claims, wherein the compound has increased capability of self-assembly.

15. The compound of any one of the preceding claims, wherein the compound is biologically active.

16. The compound of any one of the preceding claims, wherein the compound is biologically inactive.

17. The compound of claim 15, wherein the biological activity is selected from the group consisting of: anti-cancer activity, anti-microbial activity, and anti-biofilm activity.

18. The compound of claim 17, wherein the compound has an MIC against a microbial strain of less than 2.0 mM, such as from 5 pM to 1.5 mM.

19. The compound of claim 18, wherein the microbial strain is of a prokaryotic microorganism, archaea, or a eukaryotic microorganism.

20. The compound of claim 18, wherein the microbial strain is of a gram-positive bacterium or a gram-negative bacterium.

21. The compound of claim 20, wherein the gram-positive bacterium is Staphylococcus aureus (S.Aureus), for example methicillin resistant S. aureus (MRSA).

22. The compound of claim 20, wherein the gram-negative bacterium is is Escherichia coli (E. coli), Pseudomonas aeruginosa, or Acinetobacter baumannii.

23. A compound of Formula (Al):AFormula (Al),Case Ref. P319WO IPTector®wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selected from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;m is any integer, such as from 1 to 6;n is any integer, such as from 1 to 6;X is selected from the group consisting of: S, O, NH, NH2+, NR10, NHR10+, and SR10+; and wherein each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10is independently selected from the group consisting of: hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, such as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino, nitro, cyano, carboxyl, sulfonyl, phosphonyl, or fused with another one of R1, R2, R3, R4, R5, R6, R7, R8, R9, or R10of another compound of Formula (A1) forming a dimer between two compounds of Formula (A1); and wherein a pair of R1and R2, R2and R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of R1and R2, R2and R3, R3and R4, or R4and R5is bound in formula A1.

24. The compound of any one of claims 1-22, wherein the compound is of Formula (A3):Formula (A3);wherein each occurrence of Ri is independently selected from the group consisting of a lipophilic substituent and an electron-withdrawing substituent;wherein n is 1, 2, or 3,Case Ref. P319WO IPTector®for example wherein the lipophilic substituent is selected from the group consisting of C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C12 cycloalkyl, aryl, a non-aromatic ring system, a haloalkyl, such as C1-C24 haloalkyl, such as CF3, and SF5;for example wherein the electron-withdrawing substituent is selected from the group consisting of CF3, SF5, cyano, and halogen;wherein X is selected from the group consisting of O, S, guanidino, optionally substituted guanidinium, and optionally substituted thiouronium;wherein R3 is selected from the group consisting of hydrogen, C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, an aromatic ring system, and a non-aromatic ring system;wherein Y is selected from the group consisting of a phosphate, sulfate, and carboxylate; and wherein A is selected from the group consisting of a proton, and a carbon-containing cation.

25. The compound of any one of claims 1-22, wherein the compound is of Formula (A3):H H H H Formula (Cl);wherein R2 is selected from the group consisting of a lipophilic substituent and an electronwithdrawing substituent;wherein n is 1, 2, or 3,for example wherein the lipophilic substituent is selected from the group consisting of C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C12 cycloalkyl, aryl, a non-aromatic ring system, a haloalkyl, such as C1-C24 haloalkyl, such as CF3, and SF5;for example wherein the electron-withdrawing substituent is selected from the group consisting of CF3, SF5, cyano, and halogen;wherein X is selected from the group consisting of O, S, guanidino, guanidinium, and thiouronium;wherein R3 is selected from the group consisting of hydrogen, C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, an aromatic ring system, and a non-aromatic ring system;wherein Y is selected from the group consisting of a phosphate, sulfate, and carboxylate; and wherein A is selected from the group consisting of a proton, and a carbon-containing cation.

26. The compound of claim 25, wherein R2is selected from the group consisting of: C1-C24alkyl, C2- C24 alkenyl, C2-C24 alkynyl, C3-C14cycloalkyl, aryl, heteroaryl, C3-C14heterocyclyl, C C10haloalkyl, halogen, cyano, CF3, and SF5;Case Ref. P319WO IPTector®wherein each of said C1-C24alkyl, C2-C24alkenyl, C2-C24alkynyl, C3-C14cycloalkyl, aryl, heteroaryl, and C3-C14heterocyclyl groups is optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, hydroxy, -Ce alkoxy, C1-C6haloalkoxy, amino, mono(C1-C6alkyl)amino, di(C1-C6alkyl)amino, cyano, CF3, SF5, -C(=O)ORa, -C(=O)NRaRb, -SOzRa, and -SOzNRaRb;wherein any C1-C24alkyl, C2-C24alkenyl, or C2-C24alkynyl group in R2is optionally interrupted by 1 or 2 linkages independently selected from -O-, -S-, -NRa-, -C(=O)-, -C(=O)O-, -OC(=O)-, - C(=O)NRa-, and -NRaC(=O)-;wherein Ra and Rb are independently selected from H and C1-C6alkyl.

27. The compound of claim 26, wherein Rzis selected from the group consisting of: CTC12alkyl, Cz- C12alkenyl, CZ-C1Zalkynyl, C3-C10cycloalkyl, aryl, heteroaryl, -Cs haloalkyl, CF3, SF5, and cyano; wherein each of said C C12alkyl, C2-C12alkenyl, C2-C12alkynyl, C3-C10cycloalkyl, aryl, and heteroaryl groups is optionally substituted with 1 or 2 substituents independently selected from halogen, cyano, CF3, SF5, C1-C4alkoxy, -C(=O)ORa, and -C(=O)NRaRb;wherein any C1-C12alkyl, C2-C12alkenyl, or C2-C12alkynyl group in R2is optionally interrupted by 1 linkage selected from -O-, -C(=O)O-, and -C(=O)NRa-;wherein C1-C12alkyl includes linear and branched alkyl.

28. The compound of any one of claims 1-22, wherein the compound is of Formula (A3):Formula (A3);wherein each occurrence of Ri is independently selected from the group consisting of a lipophilic substituent and an electron-withdrawing substituent;wherein n is 1, 2, or 3,for example wherein the lipophilic substituent is selected from the group consisting of C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C12 cycloalkyl, aryl, a non-aromatic ring system, a haloalkyl, such as C1-C24 haloalkyl, such as CF3, and SF5;for example wherein the electron-withdrawing substituent is selected from the group consisting of CF3, SF5, cyano, and halogen;wherein X is selected from the group consisting of O, S, guanidino, optionally substituted guanidinium, and optionally substituted thiouronium;wherein R3 is selected from the group consisting of hydrogen, C1-C24 alkyl, C2-C24 alkenyl, C2-C24Case Ref. P319WO IPTector®alkynyl, an aromatic ring system, and a non-aromatic ring system;wherein Y is selected from the group consisting of a phosphate, sulfate, and carboxylate; and wherein A is selected from the group consisting of a proton, and a carbon-containing cation.

29. The compound of any one of claims 24-28, wherein A is selected from the group consisting of:a proton (H+), a substituted ammonium group, preferably tert-butyl ammonium (TBA), and a bioactive compound, for example a proton or TBA.

30. The compound of any one of claims 1-29, wherein when Y carries a negative charge, A is present in an amount that provides overall electroneutrality.

31. The compound of any one of claims 24, 28, and 29-30, wherein n is 1.

32. The compound of any one of claims 24, 28, and 29-31, wherein each occurrence of Ri is independently selected from the group consisting of: C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C14 cycloalkyl, aryl, heteroaryl, C3-C14 heterocyclyl, C1-C10 haloalkyl, halogen, cyano, CF3, and SF5;wherein each of said C1-C24 alkyl, C2-C24 alkenyl, C2-C24 alkynyl, C3-C14 cycloalkyl, aryl, heteroaryl, and C3-C14 heterocyclyl groups is optionally substituted with 1 to 3 substituents independently selected from the group consisting of halogen, hydroxy, C1-C6alkoxy, C1-C6haloalkoxy, amino, mono(C1-C6alkyl)amino, di(C1-C6alkyl)amino, cyano, CF3, SF5, -C(=O)ORa, -C(=O)NRaRb, -SO2Ra, and -SO2NRaRb;wherein any C1-C24 alkyl, C2-C24 alkenyl, or C2-C24 alkynyl group in Ri is optionally interrupted by 1 or 2 linkages independently selected from -O-, -S-, -NRa-, -C(=O)-, -C(=O)O-, -OC(=O)-, - C(=O)NRa-, and -NRaC(=O)-;wherein Raand Rbare independently selected from H and C1-C6alkyl.

33. The compound of claim 32, wherein each occurrence of Ri is independently selected from the group consisting of: C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C3-C10 cycloalkyl, aryl, heteroaryl, C1-C6haloalkyl, CF3, SF5, and cyano;wherein each of said C1-C12 alkyl, C2-C12 alkenyl, C2-C12 alkynyl, C3-C10 cycloalkyl, aryl, and heteroaryl groups is optionally substituted with 1 or 2 substituents independently selected from halogen, cyano, CF3, SF5, C1C1-C4alkoxy, -C(=O)ORa, and -C(=O)NRaRb;wherein any C1-C12 alkyl, C2-C12 alkenyl, or C2-C12 alkynyl group in Ri is optionally interrupted byCase Ref. P319WO IPTector®1 linkage selected from -O-, -C(=O)O-, and -C(=O)NRa-.

34. The compound of any one of claims 24-32, wherein R3 is selected from the group consisting of:H, C1-C24 alkyl, C2-C24alkenyl, C2-C24alkynyl, C3-C14cycloalkyl, C6-C14aryl, C7-C20aryl(C1-C6)alkyl, 5- to 14-membered heteroaryl, and 3- to 14-membered non-aromatic heterocyclyl; wherein -C^ alkyl includes linear and branched alkyl;wherein each R3other than H is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen, hydroxy, oxo, cyano, nitro, amino, mono(C1-C6alkyl)amino, di(C1-C6alkyl)amino, C C6alkyl, -Ce haloalkyl, -Ce alkoxy, C1-C6haloalkoxy, -Ce alkylthio, CF3, SF5, carboxyl, -Ce alkoxycarbonyl, -Ce alkylcarbonyl, -C(=O)NH2, -Cf^jNHfCr C6alkyl), -C(=O)N(C1-C6alkyl )2, -SO2(C C6alkyl), -SO2NH2, -SO2NH(C1-C6alkyl), -SO2N(C1-C6alkyl)2, -P(=O)(OH)2, and -P(=O)(O(C C6alkyl))2;wherein any C1-C24alkyl, C2-C24alkenyl, or C2-C24alkynyl portion of R3is optionally interrupted by 1 or 2 linkages independently selected from -O-, -S-, -NH-, -N(C1-C6alkyl)-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NH-, -NHC(=O)-, -C(=O)N(C1-C6alkyl)-, and -N(C1-C6alkyl)C(=O)-.

35. The compound of claim 34, wherein R3is selected from the group consisting of H, C1-C12alkyl, C2-C12alkenyl, C2-C12alkynyl, C3-C10cycloalkyl, phenyl, 1-naphthyl, 2-naphthyl, C7-C16aryl(C1-C4)alkyl, 5- to 10-membered heteroaryl, and 5- to 10-membered non-aromatic heterocyclyl; wherein C1-C12alkyl includes linear and branched alkyl, including isobutyl;wherein C7-C16aryl(C1-C4)alkyl includes benzyl, 1-naphthylmethyl, and 2-naphthylmethyl; wherein each R3group other than H is optionally substituted with 1, 2, or 3 substituents independently selected from the group consisting of halogen, cyano, CF3, SF5, -C4 alkyl, -C4 haloalkyl, C1-C4alkoxy, -C4 haloalkoxy, amino, mono(C1-C4alkyl)amino, di(CrC4alkyl)amino, - C(=0)0(C C4 alkyl), -C(=O)NH2, -C^OjNHfC^ alkyl), -SOzfC^ alkyl), and -SO2NH2;and wherein any alkyl, alkenyl, or alkynyl portion of R3is optionally interrupted by 1 linkage selected from -O-, -C(=O)O-, and -C(=O)NH-.

36. The compound of claim 35, wherein R3is selected from the group consisting of isobutyl, secbutyl, tert-butyl, benzyl, 1-naphthylmethyl, and 2-naphthylmethyl, each optionally substituted with 1 or 2 substituents independently selected from the group consisting of halogen, -C4 alkyl, C1-C4alkoxy, CF3, SF5, and cyano.

37. The compound of any one of claims 24, 28, and 29-36, wherein each occurrence of Ri isCase Ref. P319WO IPTector®independently selected from the group consisting of C1–C12alkyl, C2–C12alkenyl, C2–C12alkynyl, C3–C12cycloalkyl, aryl, heteroaryl, non-aromatic ring system, CF3, and SF5, cyano, nitro, halogen, C(=O)C1–C6alkyl, C(=O)O–C1–C6alkyl, C(=O)NH2, C(=O)NH(C1–C6alkyl), SO2–C1–C6alkyl, SO2NH2, and SO2NH(C1–C6alkyl).

38. The compound of any one of claims 24-37, wherein X is selected from the group consisting of O, S, guanidino, optionally substituted guanidinium, and optionally substituted thiouronium.

39. The compound of claim 23, wherein the compound is of Formula (A4);wherein R3 is isobutyl or benzyl.

40. The compound of any one of claims 1-22, wherein the compound is of Formula (Bl):R2 Formula (Bl);wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selected from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;m is any integer, such as from 1 to 6;n is any integer, such as from 1 to 6; andwherein each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10is independently selected from the group consisting of: hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, suchCase Ref. P319WO IPTector®as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino, nitro, cyano, carboxyl, sulfonyl, phosphonyl, or fused with another one of R1, R2, R3, R4, R5, R6, R7, R8, R9, or R10of another compound of Formula (B1) forming a dimer between two compounds of Formula (Bl); and wherein a pair of R1and R2, R2and R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of R1and R2, R2and R3, R3and R4, or R4and R5is bound in formula B1.

41. The compound of any one of claims 1-22, wherein the compound is of Formula (B2):AFormula (B2);wherein Y is a hydrophilic hydrogen bond acceptor group selected from: an anionic hydrophilic hydrogen bond acceptor group or a neutral hydrophilic hydrogen bond acceptor group; and if Y is an anionic hydrophilic hydrogen bond acceptor group, A is a cation, for example wherein A is selected from the group consisting of a proton (H+), an alkali metal ion (such as Na+, K+, Li+), an alkaline earth metal ion (such as Mg2+, Ca2+), ammonium (NH4+), a substituted ammonium group (preferably TBA), pyridinium, phosphonium, sulfonium, imidazolium, triazolium, guanidinium, other heterocyclic or delocalized cationic species, and an onium salt derived from nitrogen, phosphorus, or a sulfur-containing moiety, or if Y is a neutral hydrophilic hydrogen bond acceptor group, A is absent;m is any integer, such as from 1 to 6;n is any integer, such as from 1 to 6; andwherein each R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10is independently selected from the group consisting of: hydrogen, a linear or branched alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, a heterocyclic group, a haloalkyl group, such as a trifluoroalkyl group, an acyl group, an alkoxy group, an aryloxy group, an amino group, an imino group, a thioalkyl group, a thioaryl group, a silyl group, a phosphino group, a halogen atom, or any combination thereof, wherein each group may be unsubstituted or substituted with one or more substituents independently selected from halogen, hydroxyl, alkoxy, amino,Case Ref. P319WO IPTector®nitro, cyano, carboxyl, sulfonyl, phosphonyl, or fused with another one of R1, R2, R3, R4, R5, R6, R7, R8, R9, or R10of another compound of Formula (B2) forming a dimer between two compounds of Formula (B2); and wherein a pair of R1and R2, R2and R3, R3and R4, or R4and R5may be linked together to form a fused bicyclic or tricyclic ring system, wherein said fused bicyclic or tricyclic ring system includes the aryl ring to which the pair of R1and R2, R2and R3, R3and R4, or R4and R5is bound in formula B2.

42. The compound of any one of the preceding claims, wherein Y comprises a carboxylate anion.

43. The compound of any one of the preceding claims, wherein A comprises tetrabutylammonium (TBA).

44. The compound of any one of the preceding claims, wherein R6and / or R7are a hydrophobic group.

45. The compound of any one of the preceding claims, wherein one of R6and R7is a hydrophobic group and one is hydrogen.

46. The compound of any one of the preceding claims, wherein one of R6and R7is a branched or straight alkyl chain, such as isobutyl, or an aralkyl, such as benzyl.

47. The compound of any one of the preceding claims, wherein at least 2, such as at least 3, such as at least 4 of R1, R2, R3, R4, and R5is hydrogen.

48. The compound of any one of the preceding claims, wherein one of R1, R2, R3, R4, and R5is a trifluoroalkyl, such as trifluoromethyl.

49. The compound of any one of the preceding claims, wherein the compound is chiral.

50. The compound of any one of the preceding claims, wherein the compound is essentially a single enantiomer or a mixture of enantiomers, for example essentially a 1:1 mixture of enantiomers.

51. The compound of any one of the preceding claims, wherein the compound is of formula A2, A1, B1, or B2, and wherein one or more of R6, R7, R8, and R9are independently selected from H andCase Ref. P319WO IPTector® a hydrophobic group, wherein at least one of R6, R7, R8, and R9is a hydrophobic group.

52. The compound of any one of the preceding claims, wherein the compound is not any one of:Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®53. The compound of any one of the preceding claims, wherein the compound is selected from the group consisting of:Case Ref. P319WO IPTector® L118 / D119 L120 / D121 L124 / D125Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®54. The compound of any one of the preceding claims, wherein the compound is selected from the group consisting of:Case Ref. P319WO IPTector®stereoisomeric mixture thereof (SSA 72, 73, 56, 57, 74, 75).

55. The compound of any one of the preceding claims, wherein the compound is selected from the group consisting of:Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®, a stereoisomer thereof, and a salt thereof, such as a pharmaceutically acceptable salt thereof.

56. The compound of any one of the preceding claims, wherein the compound is characterized by exhibiting a membrane adhesion factor (MAF) of 0.5 or less and a membrane permeation factor (PF) of 1.0 or greater, when measured by1H CPMG NMR spectroscopy against 1:1 PE:PG phospholipid vesicles.

57. A nanostructure comprising a plurality of compounds each independently defined as in any one of claims 1-56.

58. The nanostructure of claim 57, wherein the plurality of compounds self-associate in a liquid medium, such as an aqueous medium, thereby forming the nanostructure.

59. The nanostructure of any of claims 57-58, wherein the nanostructure is configured for or capable of interacting with a biological membrane and adhering to and / or penetrating the biological membrane to form pores and / or channels in the biological membrane.

60. The nanostructure of claim 59, wherein the adhering to and / or penetrating the biological membrane increases the susceptibility of the biological membrane to a therapeutic agent and / or increases the permeability of the therapeutic agent across the biological membrane when contacted with the therapeutic agent.

61. The nanostructure of any of claims 57-60, wherein the nanostructure is capable of or configured for acting as a supramolecular host to one or more phospholipid headgroups of a biological membrane acting as supramolecular guests.

62. The nanostructure of any of claims 57-61, wherein the nanostructure is capable of forming or forms a supramolecular host:guest complex with one or more phospholipid headgroups of a biological membrane when contacted with one or more phospholipid headgroups of a biologicalCase Ref. P319WO IPTector®membrane.

63. The nanostructure of any of claims 57-62, wherein the nanostructure is capable of or configured for absorbing or adsorbing a therapeutic agent, such as a small molecule.

64. The nanostructure of any of claims 57-62, wherein the nanostructure is capable of or configured for absorbing or adsorbing a therapeutic agent, such as a small molecule, and further capable of or configured for delivering the therapeutic agent to a biological cell.

65. The nanostructure of any of claims 57-59, wherein the plurality of compounds comprises the same or different compounds.

66. The nanostructure of any of claims 57-65, wherein the plurality of compounds is selected from the group consisting of:Case Ref. P319WO IPTector®10Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®o67. The nanostructure of any of claims 57-66, wherein the plurality of compounds is selected from the group consisting of:stereoisomeric mixtures thereof.Case Ref. P319WO IPTector®68. The nanostructure of any of claims 57-67, wherein the plurality of compounds is selected from the group consisting of:stereoisomeric mixtures thereof.

69. The nanostructure of any of claims 57-68, comprising two different compounds.

70. The nanostructure of any of claims 57-69, comprising a combination of compounds selected from the group consisting of:Case Ref. P319WO IPTector®Case Ref. P319WO IPTector®71. The nanostructure of any of claims 57-70, wherein the nanostructure comprises a mixture of enantiomers, for example in a ratio of from 30 to 70, such as from 35 to 65, such as from 40 to 60, such as for example approximately 50:50.

72. The nanostructure of any of claims 57-71, wherein the plurality of compounds comprises essentially a single enantiomer.

73. The nanostructure of any of claims 57-72, wherein the nanostructure is capable of aggregating into an essentially spherical form, or a gel fiber, or a combination thereof.

74. The nanostructure of claim 73, wherein the nanostructure is essentially spherical when in a liquid medium, such as an aqueous medium.

75. The nanostructure of any one of claims 57-74, wherein the nanostructure has a hydrodynamic diameter, dHof from 90 to 600 nm, such as from 90 nm to 100 nm, such as from 100 nm to 110 nm, such as from 110 nm to 120 nm, such as from 120 nm to 130 nm, such as from 130 nm to 140 nm, such as from 140 nm to 150 nm, such as from 150 nm to 160 nm, such as from 160 nm to 170 nm, such as from 170 nm to 180 nm, such as from 180 nm to 190 nm, such as from 190 nm to 200 nm, such as from 200 nm to 210 nm, such as from 210 nm to 220 nm, such as from 220 nm to 230 nm, such as from 230 nm to 240 nm, such as from 240 nm to 250 nm, such as from 250 nm to 260 nm, such as from 260 nm to 270 nm, such as from 270 nm to 280 nm, such as from 280 nm to 290 nm, such as from 290 nm to 300 nm, such as from 300 nm to 310 nm, such as from 310 nm to 320 nm, such as from 320 nm to 330 nm, such as from 330 nm to 340 nm, such as from 340 nm to 350 nm, such as from 350 nm to 360 nm, such as from 360 nm to 370 nm, such as from 370 nm to 380 nm, such as from 380 nm to 390 nm, such as from 390 nm to 400 nm, such as from 400 nm to 410 nm, such as from 410 nm to 420 nm, such as from 420 nm to 430 nm, such as from 430 nm to 440 nm, such as from 440 nm to 450 nm, such as from 450 nm to 460 nm, such as from 460 nm to 470 nm, such as from 470 nm to 480 nm, such as from 480 nm to 490 nm, such as from 490 nm to 500 nm, such as from 500 nm to 510 nm, suchCase Ref. P319WO IPTector®as from 510 nm to 520 nm, such as from 520 nm to 530 nm, such as from 530 nm to 540 nm, such as from 540 nm to 550 nm, such as from 550 nm to 560 nm, such as from 560 nm to 570 nm, such as from 570 nm to 580 nm, such as from 580 nm to 590 nm, such as from 590 nm to 600 nm, such as from 600 nm to 610 nm, such as from 610 nm to 620 nm, such as from 620 nm to 630 nm, such as from 630 nm to 640 nm, such as from 640 nm to 650 nm, such as from 650 nm to 660 nm, such as from 660 nm to 670 nm, such as from 670 nm to 680 nm, such as from 680 nm to 690 nm, such as from 690 nm to 700 nm, such as from 700 nm to 710 nm, such as from 710 nm to 720 nm, such as from 720 nm to 730 nm, such as from 730 nm to 740 nm, such as from 740 nm to 750 nm, such as from 750 nm to 760 nm, such as from 760 nm to 770 nm, such as from 770 nm to 780 nm, such as from 780 nm to 790 nm, such as from 790 nm to 800 nm, such as from 800 nm to 810 nm, such as from 810 nm to 820 nm, such as from 820 nm to 830 nm, such as from 830 nm to 840 nm, such as from 840 nm to 850 nm, such as from 850 nm to 860 nm, such as from 860 nm to 870 nm, such as from 870 nm to 880 nm, such as from 880 nm to 890 nm, such as from 890 nm to 900 nm, such as from 900 nm to 910 nm, such as from 910 nm to 920 nm, such as from 920 nm to 930 nm, such as from 930 nm to 940 nm, such as from 940 nm to 950 nm, such as from 950 nm to 960 nm, such as from 960 nm to 970 nm, such as from 970 nm to 980 nm, such as from 980 nm to 990 nm, such as from 990 nm to 1000 nm.

76. The nanostructure of any one of claims 57-75, wherein the nanostructure has a hydrodynamic diameter (dH) of less than 250 nm, for example from 10 nm to 249 nm, for example from 120 nm to 250 nm.

77. The nanostructure of any one of claims 57-76, wherein the nanostructure is characterized by having a polydispersity index (PDI) of less than 0.1, such as from 0.001 to 0.09, for example wherein the nanostructure has a polydispersity index (PDI) of less than 0.05.

78. The nanostructure of any one of claims 57-77, wherein the nanostructure is characterized by having a Zeta potential of from -20 to -100 mV, for example from -40 to -90 mV or from -70 mV to -30 mV.

79. The nanostructure of any one of claims 57-78, wherein the nanostructure is characterized by a Critical Aggregation Concentration (CAC) of less than 85 mM.

80. The nanostructure of any one of claims 57-79, wherein the nanostructure is characterized byCase Ref. P319WO IPTector®having a Critical Aggregation Concentration (CAC) of from 0.3 mM to 10 or above mM, for example from 0.5 mM to 10 mM.

81. The nanostructure of any one of claims 57-80, wherein the nanostructure is characterized by a surface tension (ST) at the Critical Aggregation Concentration (CAC) of less than 50 mN-m-1, for example wherein the surface tension (ST) at the Critical Aggregation Concentration (CAC) is less than 45 mN-m-1.

82. The nanostructure of any one of claims 57-80, wherein the nanostructure is characterized by having a surface tension (ST) at the Critical Aggregation Concentration (CAC) of from 30 to 55 mNm1, such as from 40 to 50 mNm-1.

83. The nanostructure of any one of claims 57-80, wherein the nanostructure is characterized by having a K of from 2.0 to 9000.0 M-1, such as from 3.0 to 8000.0 M-1.

84. The nanostructure of any one of claims 57-83, wherein the nanostructure is capable of or configured for forming pores and / or channels in a biological cell membrane, for example as evidenced by patch clamp analysis.

85. The nanostructure of claim 84, wherein the formation of pores and / or channels is evidenced by the induction of one or more ion transport events corresponding to a current of at least 0.1 x 10−11A, for example at least 5.00 x 10−11A when the nanostructure is applied to a planar phospholipid bilayer membrane at a concentration of 1.0 mM or less with respect to the compound and at a holding voltage of +100 mV.

86. The nanostructure of claim 84, wherein the ion transport events correspond to a current of from 0.1 x 10-11A to 5.0 x 10“8A, for example wherein the ion transport events correspond to a current of from 1.00 x 10“10A to 1.00 x 10“8A.

87. The nanostructure of claim 84, wherein the ion transport events correspond to a current of from 1.00 x IO’10A to 1.00 x 10“9A.

88. A pharmaceutical composition comprising a compound as defined in any one of claims 1-56, or a nanostructure as defined in any one of claims 57-87, and one or more pharmaceuticallyCase Ref. P319WO IPTector®acceptable excipients.

89. The pharmaceutical composition of claim 88, wherein the composition further comprises a solvent.

90. The compound, nanostructure, or pharmaceutical composition of any one of the preceding claims, further comprising a therapeutic agent.

91. The compound, nanostructure, or pharmaceutical composition according to claim 90, wherein the therapeutic agent is a small molecule.

92. The compound, nanostructure, or pharmaceutical composition according to any of claims 90- 91, wherein the therapeutic agent is selected from the group consisting of: cisplatin, olaparib, gemcitabine, paclitaxel, and mitomycin C.

93. The compound, nanostructure, or pharmaceutical composition according to any of claims 90- 92, wherein the therapeutic agent is selected from the group consisting of: cisplatin, olaparib and paclitaxel.

94. The compound, nanostructure, or pharmaceutical composition according to any of claims 61- 62, wherein the therapeutic agent is selected from the group consisting of: olaparib and paclitaxel.

95. A phospholipid membrane comprising the compound as defined in any one of claims 1-56, or the nanostructure as defined in any one of claims 57-87.

96. The phospholipid membrane of claim 95, wherein the membrane has been coated, penetrated, and / or disrupted by the nanostructure.

97. The phospholipid membrane of any of claims 95-96, wherein the membrane is naturally occurring or synthetic.

98. The phospholipid membrane of any of claims 95-97, wherein the nanostructure enables cation transport across the phospholipid membrane, for example wherein the cation is a sodium ionCase Ref. P319WO IPTector®or a potassium ion.

99. The phospholipid membrane of any of claims 95-97, wherein the nanostructure enables exchange of two different ions or molecules across the membrane in opposite directions.

100. The phospholipid membrane of any of claims 95-98, wherein the membrane further comprises an anionophore.

101. A cell comprising the membrane as defined in any one of claims 95-100.

102. A method for treatment of a pathological condition in a subject, the method comprising administering a compound, a nanostructure, or a pharmaceutical composition to the subject.

103. The method of claim 72, wherein the compound is as defined in any one of claims 1-56, the nanostructure is as defined in any one of claims 57-87, or the pharmaceutical composition is as defined in claim 88.

104. The method of claim 102, wherein the pathological condition is a disease.

105. The method of any one of claims 102-104, wherein the method further comprises administering, simultaneously or sequentially, a therapeutic agent to the subject.

106. The method of claim 105, wherein the ratio of the therapeutic agent to the compound is from 1:5 to 1:20.

107. The method of any one of claims 102-106, wherein the compound, nanostructure or pharmaceutical composition is administered to the subject orally, intraveneously, or intravesicularly.

108. A compound, a nanostructure, or a pharmaceutical composition for use in the treatment of a pathological condition in a subject, the use comprising administering the compound, the nanostructure, or the pharmaceutical composition to the subject.

109. The method of any one of claims 102-106, wherein the pathological condition is a cancer.Case Ref. P319WO IPTector®110. The methof claim 109, wherein the cancer is selected from the group consisting of: bladder cancer, and ovarian cancer.

111. The method of any of claims 109-110, wherein the compound, the nanostructure, or the pharmaceutical composition comprises compound (SSA48):ss,O’ TBA(SSA48).

112. The method of any one of claims 102-105, wherein the pathological condition is a microbial infection.

113. The method of of claim 112, wherein the microbial infection has resulted from a gram-positive bacterium or a gram-negative bacterium.

114. The method of claim 113, wherein the gram-positive bacterium is Staphylococcus aureus (S.Aureus), for example methicillin resistant S. aureus (MRSA).

115. The method of claim 113, wherein the gram-negative bacterium is Escherichia coli (E. coli).

116. A method for inhibiting biofilm on a surface of an object, the method comprising administering a compound or a nanostructure to the surface of the object.

117. The method of claim 116, wherein the compound is defined as in any one of claims 1-56, or the nanostructure is defined as in any one of claims 57-87.

118. A process for preparing a nanostructure comprising a plurality of compounds each independently defined as in any one of claims 1-56, the process comprising:a. providing a plurality of compounds each independently defined as in any one of claims 1-56; b. subjecting the compounds to a liquid medium, such as an aqueous medium;c. thereby obtaining the nanostructure.

119. A method of selecting a compound capable of or suitable for forming pores and / or channels in a bilayer membrane, for example a phospholipid bilayer membrane, such as a biological cellCase Ref. P319WO IPTector®membrane of a cell, the method comprising one or more or all of:e) Identifying a compound capable of complimentary interacting with phospholipid groups of a biological membrane of a cell;f) Identifying a compound engaging in self-associative hydrogen bonding events;g) Selecting a compound as defined in any one of claims 1-56;h) Subjecting a compound to patch clamp analysis to confirm presence of formation of pores and / or channels in the biological cell membrane,thereby selecting the compound suitable for or capable of forming pores and / or channels in the bilayer membrane.

120. The method of claim 119, wherein the method is performed in vitro, ex vivo, or in vivo.

121. The method of any of claims 119-120, wherein the patch clamp analysis comprises subjecting one or more concentrations of a compound, for example a compound as defined in any one of claims 1-34, in one or more experiments to patch clamp analysis, applying an external voltage and after a course of time categorizing time spent in each of a number of event states.

122. The method of any of claims 119-121, wherein the patch clamp analysis comprises subjecting one or more concentrations of a compound to a bilayer membrane which is a phospholipid membrane.

123. The method of any of claims claims 119-122, wherein the number of event states are categorised as follows (current, A) 0: -co A < Event 0 < 5.00e-ll A, 1: 5.00e-ll A < Event 1 < 5.00e-10 A, 2: 5.00e-10 A < Event 2 < 4.90e-08 A, 3: 4.90e-08 A < Event 3 < co.

124. The method of any of claims 121-123, wherein the external voltage is from +70 mV to +200 mv, for example +100 mV.

125. The method of any of claims 121-124, wherein the course of time is from 400 seconds to 2000 seconds, such as approximately 1000 seconds.

126. The method of any of claims 121-125, further comprising measuring the change in current (A) over time and detecting one or more increases in current over time.Case Ref. P319WO IPTector®127. The method of any of claims 121-126, wherein a) detecting one or more increases in current over time and / or b) detecting at least 10% presence, such as at least 20% presence, such as at least 30% presence, such as at least 40% presence in more than one event states over the course of time confirms the compound being capable of or suitable for forming pores and / or channels in the biological cell membrane.

128. A method of forming pores and / or channels in a biological cell membrane of a cell using a nanostructure as defined in any one of claims 57-84, the method comprising contacting the nanostructure with the biological cell membrane of the cell in a liquid medium, for example an aqueous medium, thereby allowing the nanostructure to adhere to and permeate the biological cell membrane forming pores and / or channels.

129. A method of using a compound as defined in any one of claims 1-56 to deliver a therapeutic agent to a biological cell, comprising the steps:a. mixing the compound with the therapeutic agent in a liquid medium;b. forming a nanostructure comprising the compound and the therapeutic agent;c. delivering the nanostructure to a mammal comprising the biological cell, andd. allowing the nanostructure to adhere to and penetrate the biological cell.

130. The method of any of claims 128-129, wherein the biological cell is a pathogenic cell, such as a cancer cell or a microbial cell.

131. The method of any of claims 128-130, wherein the nanostructure forms dynamic pores and / or channels.

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