Targeted dynamic antimicrobial systems

Abstract

The invention relates to a molecular system comprising: (i) at least one dynamic constitutional framework (DCF) comprising a plurality of cores and a plurality of connectors, (ii) a plurality of functional heads, (iii) at least one pillararene and, optionally, (iv) at least one anti-infective agent; wherein each of the anti-infective agents is hosted in one of the pillararenes; wherein the cores, connectors, functional heads, and pillararenes are covalently bound together by reversible bonds; and wherein each of the cores is covalently bound to at least three from the connectors, functional heads and pillararenes. The invention also relates to a pharmaceutical composition or medicament comprising the molecular system. The invention also relates to the molecular system for use in the treatment of an infectious disease such as a bacterial disease, preferably a disease caused by Acinetobacter baumannii bacteria.

Description

TARGETED DYNAMIC ANTIMICROBIAL SYSTEMSFIELD OF INVENTION

[0001] The present invention relates to a molecular system useful as “targeted dynamic antimicrobial system”, in particular for use as anti-infective agent, especially as antibacterial agent.BACKGROUND OF INVENTION

[0002] Acinetobacter baumannii is a Gram-negative bacterium ranked as top critical priority by both WHO and CDC. Resistance to last-resort antibiotics earned this bacterium a place amongst the most problematic nosocomial ESKAPE pathogens. Acinetobacter baumannii is well-known for forming biofilms, complex communities of microbial cells embedded in a selfgenerated extracellular polymeric matrix. They can either be surface-attached or occur as non¬ surface-attached aggregates, which poses an even more significant threat in healthcare settings, since the microorganisms within biofilms are shielded from the effects of antibiotics, necessitating ongoing efforts to develop novel compounds with antibiofilm activity. New alternative therapies are therefore urgently required to combat this increasingly resistant pathogen. Classical strategies employed by medicinal chemists to develop new antibiotics have been impaired by both the lack of novel central scaffolds and the rapid emergence of bacterial resistance. Moreover, the concept of personalized medicine is rapidly emerging, including for treating infectious diseases, thus opening the search for novel personalized antibacterial agents. The latter would lower the side effects, decrease or delay the resistance processes, and preserve the host microbiota, three critical features in developing novel antibacterial therapies.

[0003] It was surprisingly found out that a molecular system comprising a Dynamic Constitutional Framework (DCF) functionalized by functional heads and at least one anti-infective agent hosted in a pillararene can be a potent anti-infective agent, especially a potent antimicrobial agent (e.g., antibiofilm agent). In particular, significant synergistic anti-infective effect within the molecular system has been evidenced. The molecular system is especially useful when directed towards Acinetobacter baumannii bacteria.SUMMARY

[0004] This invention relates to a molecular system comprising: (i) at least one dynamic constitutional framework (DCF) comprising: a plurality of cores, wherein each core is independently selected from phenyl, monocyclic heteroaryl, polycyclic aryl, and polycyclic heteroaryl; and a plurality of connectors, wherein each connector is a bifunctional linear homopolymer; wherein each of the connectors is covalently bound to two of the cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product; (ii) a plurality of functional heads, wherein each functional head is independently selected from amines, polyamines, hydrazides, and acyl-hydrazides; wherein each of the functional heads is covalently bound to one of the cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product; (iii) at least one pillararene; and (iv) at least one anti -infective agent; wherein each of the anti-infective agents is hosted in one of the pillararenes; and wherein each of the pillararenes is covalently bound to at least one of the cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product; wherein each of the cores is covalently bound to at least three groups selected from the connectors, the functional heads, and the pillararenes.

[0005] According to some embodiments, each core is a trivalent phenyl, preferably an 1,3,5-subsituted phenyl. According to some embodiments, each connector is a polyethylene glycol (PEG) terminated by two (C1-C4) alkyls or polypropylene glycol (PPG) terminated by two (C1-C4) alkyls, preferably a polyethylene glycol (PEG) terminated by two propyls, more preferably a polyethylene glycol (PEG) terminated by two n-propyls.

[0006] According to some embodiments, the functional heads are selected from the formulae (A) to (W) herein; wherein one of the -NH2 groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the functional head is covalently bound to one of the cores. In some embodiments, the functional heads are selected from formulae (B’) and (C’) herein; wherein one of the -NH2 groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the functional head iscovalently bound to one of the cores; and wherein Mn is 2000 Da in the compound of formula (B’).

[0007] According to some embodiments, the pillararene is a pillar[5]arene, preferably selected from the formulae (APAF) and (APA2’) herein; wherein at least one of the -NH2or -NH3+groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the pillararene is covalently bound to at least one of the cores.

[0008] According to some embodiments, the anti-infective agent is an antibiotic agent; preferably the antibiotic agent is selected from levofloxacin (Lev), lincomycin (Lin) and cefalexin (Cef); more preferably the antibiotic agent is levofloxacin (Lev).

[0009] According to some embodiments, each reversible coupling product in the molecular system is independently selected from N=CH, S-S, S(C=O)O, C(=O)O, (C=O)O(C=O), O, S, NHC(NH2+)CH2, NHNH=CH, C=C, C=C, and (C=O)NHN=CH. In some embodiments, reversible coupling product in the molecular system is N=CH.

[0010] According to some embodiments, the molecular system further comprises at least one antibody; preferably at least one immunoglobulin single variable domain (ISVD), such as a Variable Heavy domain of Heavy chain (VHH), or single domain antibody (sdAb) and / or preferably the antibody specifically binds to a cell surface of bacteria of the genus Acinetobacter, more preferably to a cell surface of bacteria of the species Acinetobacter baumannii, furthermore preferably to the protein Omp25 from Acinetobacter baumannii.

[0011] This invention also relates to a pharmaceutical composition comprising a molecular system according to the invention and at least one pharmaceutically acceptable carrier. This invention also relates to a molecular system according to the invention or the pharmaceutical composition according to the invention for use as a medicament. This invention also relates to a molecular system according to the invention or the pharmaceutical composition according to the invention for use in the treatment of an infectious disease, preferably a bacterial disease, more preferably a bacterial disease caused by Acinetobacter baumannii bacteria.

[0012] This invention also relates to a process for manufacturing a molecular system according to the invention, wherein the process comprises the following steps: (a) a step ofreacting a plurality of cores as defined in the invention with a plurality of connectors as defined in the invention, thereby obtaining a base dynamic constitutional framework (base DCF); then (b) a step of reacting the base DCF with a plurality of precursors of a functional head as defined in the invention, thereby obtaining a functionalised dynamic constitutional framework (DCF); (c) providing a host-guest complex comprising at least one pillararene as defined in the invention and at least one anti-infective agent as defined in the invention, wherein each of the anti-infective agent(s) is hosted in one of the pillararene(s); (d) a step of reacting the functionalised dynamic constitutional framework (DCF) with the host-guest complex, thereby obtaining the molecular system according to the invention; and (e) optionally, contacting and / or reacting the molecular system with a plurality of immunoglobulin single variable domains (ISVDs), thereby obtaining the molecular system according to the invention.

[0013] This invention also relates to a hosting system comprising: (i) at least one dynamic constitutional framework (DCF) comprising a plurality of cores and a plurality of connectors, as defined in the invention; (ii) a plurality of functional heads as defined in the invention; wherein each of the functional heads is covalently bound to one of the cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product; (iii) at least one pillararene; wherein each of the pillararenes is covalently bound to at least one of the cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product; wherein each of the cores is covalently bound to at least three groups selected from the connectors, the functional heads, and the pillararenes and wherein each of the pillararenes does not host any anti-infective agent.DEFINITIONS

[0014] In the present invention, the following terms have the following meanings.Chemical definitions

[0015] Where chemical substituents are combinations of chemical groups, the point of attachment of the substituent to the molecule is by the last chemical group recited on the right of the name of the substituent. For example, an arylalkyl substituent is linked to the rest of the molecule through the alkyl moiety and it may by represented as follows: “aryl-alkyl-”. Unlessotherwise indicated, the compounds were named using ChemDraw Professional 23.0 (Revvity Signals).

[0016] “Acyl-hydrazide” refers to a compound of general formula R-C(O)-HN-NH-R’, wherein R and R’ represent independently a substituent such as, for example, alkyl or aryl. Typically, “acyl-hydrazide” refers to a compound including the -C(O)-HN-NH2group (z.e., R’ represents H), namely, an “acyl hydrazone”.

[0017] “Alkyl” refers to a saturated linear or branched hydrocarbon chain, typically comprising from 1 to 16 carbon atoms, preferably from 1 to 12 carbon atoms, more preferably from 1 to 8 carbon atoms, furthermore preferably from 1 to 6 carbon atoms, furthermore preferably from 1 to 4 carbon atoms. Alkyl groups may be monovalent or polyvalent (z.e., “alkylene” groups, which are divalent alkyl groups, are encompassed in “alkyl” definition). Non-limiting examples of alkyl groups include methyl, ethyl, zz-propyl, z'-propyl, zz-butyl, z'-butyl, 5-butyl and -butyl, pentyl and its isomers (e.g., zz-pentyl, z'so-pentyl), and hexyl and its isomers (e.g., zz-hexyl, z'so-hexyl). Particular examples of alkyl groups include methyl, ethyl, zz-propyl, z'-propyl, zz-butyl, -butyl and -butyl (including methylene, ethylene, zz-propylene, zz-butyl ene and zz-butylene).

[0018] “Amine” refers to derivatives of ammonia (NH3), wherein one or more hydrogen atoms have been replaced by a substituent such as, for example, alkyl or aryl. Preferably “amine” refers to refers to a compound including the -NH2 group (“amino” group).

[0019] “Aryl” refers to a cyclic, polyunsaturated, aromatic hydrocarbyl group comprising at least one aromatic ring and comprising from 5 to 12 carbon atoms, preferably from 6 to 10 carbon atoms. Aryl groups may be monovalent or polyvalent (e.g., divalent). Aryl groups may have a single ring (e.g., phenyl) or multiple aromatic rings fused together (e.g., naphthyl) or linked covalently. The aromatic ring may optionally include one to two additional rings (either cycloalkyl, heterocycloalkyl or heteroaryl) fused thereto. This definition of “aryl” encompasses the partially hydrogenated derivatives of the carbocyclic systems enumerated herein, as long as at least one ring is aromatic. Non-limiting examples of aryl groups include phenyl, biphenyl, biphenylenyl, 5- or 6-tetralinyl, naphthalen-1- or -2-yl, 4-, 5-, 6 or 7-indenyl, 1- 2-, 3-, 4- or 5-acenaphthylenyl, 3-, 4- or 5-acenaphthenyl, 1- or 2-pentalenyl, 4- or 5-indanyl, 5-, 6-, 7- or 8-tetrahydronaphthyl, 1,2,3,4-tetrahydronaphthyl, 1,4-dihydronaphthyl, and 1-, 2-, 3-, 4- or 5-pyrenyl. A particular example of aryl group is phenyl.

[0020] “Cx-Cy” or “(Cx-Cy)” preceding the name of a group means that the group comprises from x to y carbon atoms, in accordance to common terminology in the chemistry field.

[0021] “Coupling function” refers to a function capable to react with another function to form a covalent linkage, z.e., is covalently reactive under suitable reaction conditions, and generally represents a point of attachment for another substance. The coupling function is a moiety on a first compound that is capable of chemically reacting with a functional group on a second different compound to form a covalent linkage. Coupling functions generally include nucleophiles, electrophiles and photoactivatable groups. Non-limiting examples of coupling functions are amine (e.g., amino), isothiocyanate (-N=C=S), isocyanate (-N=C=O), activated ester (e.g., N-hydroxysuccinimide ester, N-hydroxyglutarimide ester or maleimide ester), carboxylic acid, activated carboxylic acid (e.g., acid anhydride or acid halide), alcohol, alkene, ran -cyclooctene (TCO), alkyne, bicyclo[6.1.0]nonyne (BCN), dibenzocyclooctyne (DBCO), halide (e.g., Br or Cl), azide (-N3), siloxy, phosphonic acid, thiol, tetrazine, norbornen, oxoamine, aminooxy (-O-NH2), thioether, haloacetamide, glutamate, glutaric anhydride, succinic anhydride, maleic anhydride, aldehyde, ketone, hydrazide, chloroformate (-C(=O)O-Cl) and maleimide.

[0022] “Coupling product” refers to the residue of a coupling function resulting from the reaction between two coupling functions, such as, for example, a functionally related group of atoms (e.g., amide -C(O)-NH- group or a double bond) or a heterocycle (e.g., a divalent triazolyl group). For example, reaction between two coupling functions A and B may lead to the following coupling products as shown below, wherein X represents a halogen atom (e.g., Br or Cl).Coupling function A Coupling function B Coupling productR-N3 R-C=CH Triazolyl (divalent)R-NH2 R’-COOH R-NH(C=O)-R’R-SH R’-SH R-S-S-R’R-OH R’ -(epoxide group) R-OCH2CH(OH)-R’ R-NH2 R’ -(epoxide group) R-NHCH2CH(OH)-R’ R-SH R’ -(epoxide group) R-SCH2CH(OH)-R’ R-NH2 R’-CHO R-N=CH-R’ R-N=CH-R’” R-N=CH-R’ R”-N=CH-R’”R”-N=CH-R’ R-NH2 R’-NCO R-NH(C=O)NH-R R-NH2 R’-NCS R-NH(C=S)NH-R’ R-SH R’-(C=0)CH3 R-(C=O)CH2S-R’ R-SH R’-O(C=O)X R-S(C=O)O-R’ R-CH=CH2R’-SH R-CH2CH2S-R’ R-SH R’-SH R-S-S-R’R-S-S-R’” R-S-S-R’ R”-S-S-R’”R”-S-S-R’ R-SeH R’-SeH R-Se-Se-R’ R-Se-Se-R’” R-Se-Se-R’ R”-Se-Se-R’”R”-Se-Se-R’ R-B(OCHCH2O)-R’ R-B(OH)2R’-CH(OH)CH2OH (1,3,2-dioxaborolane, divalent) R-B(OH)22 R’-0H R’-O-B(R)-O-R’R’-B(OH)2R-Boroxine(R’)-R” R-B(OH)2R”-B(OH)2(boroxine, trivalent)(R3)Si-O-Si(R’”3) (R3)Si-O-Si(R’3) (R”3)Si-O-Si(R’”3)(R”3)Si-O-Si(R’3) R-OH R’-NCO R-O(C=O)NH-R’ R-SH R’-(C=O)CH2X R-SCH2(C=O)-R’ R-NH2 R’-(C=O)N3R-NH(C=O)-R’ R-COOH R’-OH R-C(=O)O-R’R-COOH R’-COOH R-(C=O)O(C=O)-R’R-OH R’-X R-O-R’ R-SH R’-X R-S-R’ R-NH2 R-CH2C(NH2+)OCH3R-NHC(NH2+)CH2-R’R-NHNH2 R’-CHO R-NHNH=CH-R’R-CH=CH-B(OH)2R’-X R-CH=CH-R’R-C=CH R’-X R-C=C-R’ R-(C=O)NHNH2R’-CHO R-(C=O)NHN=CH-R’ R-O-NH2 R’-CHO R-O-N=CH-R’

[0023] For example, a coupling product can be selected from N=CH, ON=CH, S-S, S-CH2, Se-Se, B-O, B(O)-O, Si-O, Si-O-Si S(C=O)O, (C=O)O, (C=O)O, (C=O)O(C=O), O, S, NHC(NH2+)CH2, NHN=CH, C=C, C=C; and (C=O)NHN=CH.

[0024] In some embodiments, reaction between two coupling functions A and B may lead to the following coupling products as shown below, wherein X represents a halogen atom (e.g., Br or Cl).Coupling function A Coupling function B Coupling productR-N3 R-C=CH Triazolyl (divalent)R-NH2 R’-COOH R-NH(C=O)-R’R-SH R’-SH R-S-S-R’ R-OH R’ -(epoxide group) R-OCH2CH(OH)-R’R-NH2 R’ -(epoxide group) R-NHCH2CH(OH)-R’R-SH R’ -(epoxide group) R-SCH2CH(OH)-R’R-NH2 R’-CHO R-N=CH-R’ R-NH2 R’-NCO R-NH(C=O)NH-RR-NH2R’-NCS R-NH(C=S)NH-R’R-SH R’-(C=O)CH3 R-(C=O)CH2S-R’R-SH R’-O(C=O)X R-S(C=O)O-R’R-CH=CH2R’-SH R-CH2CH2S-R’ R-OH R’-NCO R-O(C=O)NH-R’R-SH R’-(C=O)CH2X R-SCH2(C=O)-R’R-NH2R’-(C=O)N3 R-NH(C=O)-R’R-COOH R’-OH R-C(=O)O-R’ R-COOH R’-COOH R-(C=O)O(C=O)-R’R-OH R’-X R-O-R’ R-SH R’-X R-S-R’ R-NH2R-CH2C(NH2+)OCH3R-NHC(NH2+)CH2-R’R-NHNH2R’-CHO R-NHNH=CH-R’R-CH=CH-B(OH)2R’-X R-CH=CH-R’R-C=CH R’-X R-C=C-R’ R-(C=O)NHNH2R’-CHO R-(C=O)NHN=CH-R’

[0025] For example, a coupling product can be selected from triazolyl, NH(C=O), S-S, OCH2CH(OH), NHCH2CH(OH), SCH2CH(OH), N=CH, NH(C=O)NH, NH(C=S)NH, (C=O)CH2S, S(C=O)O, CH2CH2S, O(C=O)NH, SCH2(C=O), NH(C=O), C(=O)O, (C=O)O(C=O), O, S, NHC(NH2+)CH2, NHNH=CH, C=C, C=C, and (C=O)NHN=CH.

[0026] Especially, a coupling product can be selected from triazolyl, NH(C=O), S-S, OCH2CH(OH), NHCH2CH(OH), SCH2CH(OH), N=CH, NH(C=O)NH, NH(C=S)NH, (C=O)CH2S, S(C=O)O, CH2CH2S, O(C=O)NH, SCH2(C=O), NH(C=O), C(=O)O, (C=O)O(C=O), O, S, NHC(NH2+)CH2, NHNH=CH, C=C, C=C, and (C=O)NHN=CH; or selected from N=CH, ON=CH, S-S, S-CH2, Se-Se, B-O, B(O)-O, Si-O, Si-O-Si S(C=O)O,(C=0)0, (C=0)0, (C=O)O(C=O), O, S, NHC(NH2+)CH2, NHN=CH, C=C, C=C; and (C=O)NHN=CH.

[0027] “Dynamic constitutional framework” or “DCF” refers to adaptive 3D dynamic polymeric-dynamic networks that form and rearrange their structure based on reversible covalent or non-covalent interactions. These frameworks exhibit dynamic behaviour, allowing them to self-organize, self-heal, and adapt to external stimuli, making them highly versatile in various applications.

[0028] “Heteroaryl” refers to aromatic rings or aromatic ring systems comprising from 5 to 15 carbon atoms, preferably from 4 to 12 carbon atoms, more preferably from 3 to 10 carbon atoms, having one or two rings that are fused together or linked covalently, wherein at least one ring is aromatic, and wherein one or more carbon atoms in one or more of these rings is replaced by oxygen, nitrogen and / or sulphur atoms. Heteroaryl groups may be monovalent or polyvalent (e.g., divalent). The nitrogen and sulphur heteroatoms may optionally be oxidized and the nitrogen heteroatoms may optionally be quatemized (e.g., the heteroatom is substituted by oxo (=0) for sulphur atom or (— >0) for nitrogen atom). This definition of “heteroaryl” encompasses the partially hydrogenated derivatives of the carbocyclic systems enumerated herein, as well as ring systems including one or more fused non-aromatic cycloalkyl and / or heterocycloalkyl ring(s), as long as at least one ring is aromatic. The heteroaryl can be bound to another group or molecule through a carbon atom, i.e., the binding atom is not selected among the heteroatoms included therein. Alternatively, the heteroaryl can be bound to another group or molecule through one of the heteroatoms included therein. When substituted by one or more other group(s), an heteroaryl may be substituted either through a carbon atom or through a heteroatom (e.g., nitrogen), unless otherwise specified. Non-limiting examples of heteroaryl groups include pyrrolyl, furanyl, thiophenyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, oxatriazolyl, thiatriazolyl, pyridinyl, pyrimidyl, pyrazinyl, pyridazinyl, oxazinyl, dioxinyl, thiazinyl, triazinyl, tetrazinyl, imidazo[2,l-b][l,3]thiazolyl, thieno[3,2-b]furanyl, thieno[3,2- b]thiophenyl, thieno[2,3-d][l,3]thiazolyl, thieno[2,3-d]imidazolyl, tetrazolo[l,5-a]pyridinyl, indolyl, indolizinyl, isoindolyl, benzofuranyl, isobenzofuranyl, benzothiophenyl, isobenzothiophenyl, indazolyl, benzimidazolyl, 1,3-benzoxazolyl, 1,2-benzisoxazolyl, 2,1-benzisoxazolyl, 1,3-benzothiazolyl, 1,2-benzoisothiazolyl, 2,1 -benzoisothiazolyl,benzotri azolyl, 1,2,3-benzoxadiazolyl, 2,1,3-benzoxadiazolyl, 1,2,3-benzothiadiazolyl, 2.1.3-benzothiadiazolyl, thienopyridinyl, purinyl, imidazo[l,2-a]pyridinyl, 6-oxo-pyridazin-l(6H)-yl, 2-oxopyridin-l(2H)-yl, 6-oxo-pyridazin-l(6H)-yl, 2-oxopyridin-l(2H)-yl, 1,3-benzodioxolyl, quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl and quinoxalinyl. Nonlimiting examples of heteroaryl groups comprising at least one fused non-aromatic ring include 2.3 -dihydrobenzofuranyl, benzo[d][1,3]dioxolyl, indolinyl, 2,3 -dihydrobenzo / Z> ]\_ 1,4]dioxinyl, 3,4-dihydro-2 7-benzo / Z> ]\ 1,4]oxazinyl, 1,2,3,4-tetrahydroquinoxaline, 3,4-dihydro-2H-benzo[b][l,4]thiazine and 2.3-dihydrobenzo[b][l,4]oxathiine.

[0029] “Hydrazide” refers to a compound of general formula R-HN-NH-R’, wherein R and R’ represent independently a substituent such as, for example, alkyl or aryl. Preferably, “hydrazide” refers to a compound including the -HN-NH2group (z.e., R’ represents H).

[0030] “Pillararene” refers to macrocycles with a main scaffold consisting of hydroquinone or dialkoxybenzene units (typically from 5 to 10) linked in the para position by methylene (-CH2-) bridges. A “ pillar [x] arene” is a pillararene composed of x hydroquinone or dialkoxybenzene units.

[0031] “Polyamine” is an amine comprising at least two -NH- groups, typically from 20 to 2400 -NH- groups, preferably at least 70 -NH- groups.

[0032] “Prodrug” refers to a pharmacologically acceptable derivative of a therapeutic agent (e.g., a compound according to the invention) whose in vivo biotransformation product is the therapeutic agent (active drug). Prodrugs are typically characterized by increased bioavailability and are readily metabolized in vivo into the active compounds. Non-limiting examples of prodrugs include amide prodrugs and carboxylic acid ester prodrugs.

[0033] “Reversible coupling product” refers to coupling product as defined herein, which is in equilibrium with original starting materials upon the particular condition and can participate in certain equilibrium-controlled reactions (dissociation, hydrolysis, exchange, metathesis). Typically, a reversible coupling product (e.g, imine) participates in three types of reactions: hydrolysis (i.e., the coupling product reverts back to the original compound(s) by addition of water), exchange i.e., upon introduction of a compound with a coupling function, the originalcoupling product may undergo a reaction where the substituents are exchanged), and metathesis (z'.e., upon introduction of a second coupling product, the two coupling products can undergo a reaction in which the substituents are exchanged). In particular, imine participates in three types of reactions: imine hydrolysis (z.e., the imine reverts back to the original compound(s) containing amino and carbonyl groups by addition of water), imine exchange (z'.e., upon introduction of a second amine, the original imine may undergo transamination where the substituents are exchanged), and imine metathesis (z'.e., upon introduction of a second imine, the two imines can undergo a reaction in which the substituents are exchanged). In particular, this reversibility is observed in aqueous conditions such as, for example, wherein pH is about 7 and temperature is about 25°C. This reversibility can also be observed physiological conditions such as, for example, wherein pH is about 7.4 and temperature is about 37°C. For example, a reversible coupling product can be selected from N=CH, S-S, S(C=O)O, C(=O)O, (C=O)O(C=O), O, S, NHC(NH2+)CH2, NHNH=CH, C=C, C=C, and (C=O)NHN=CH, preferably N=CH.

[0034] “Solvate” refers to molecular complex comprising a compound along with stoichiometric or sub-stoichiometric amounts of one or more molecules of one or more solvents, typically the solvent is a pharmaceutically acceptable solvent such as, for example, ethanol. The term “hydrate” refers to a solvate when the solvent is water (H2O).General definitions

[0035] “About” is used herein to mean approximately, roughly, around, or in the region of. The term “about” preceding a figure means more or less 10 % of the value of the figure. When the term “about” is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth by 10%.

[0036] “Administration”, or a variant thereof (e.g., “administering”), means providing the active agent or active ingredient, alone or as part of a pharmaceutically acceptable composition, to the patient in whom / which the condition, symptom, or disease is to be treated or prevented.

[0037] “Antibiotic agent” (in short “antibiotic”) and “antibacterial agent” are synonyms and refer to a anti-infective agent as defined herein, which acts against bacteria.

[0038] “Antibiofilm” refers to the ability of a compound or substance to effectively prevent the formation and / or growth of complex microorganism communities called biofilms. Biofilms may be the cause of infectious diseases because they are very commonly found in human environment. Biofilms often present higher drug resistance than isolated microorganisms.

[0039] “Anti-infective agent” refers to a compound or substance that acts against pathogen microorganisms either by inhibiting their dissemination or growth, or directly by killing them. This generic term covers antimicrobials (including antibacterial or antibiotic, antifungal and antimycobacterial agents) and antiviral agents. Anti-infective agents (in short “anti-infectives”) are useful in the treatment of infections or infectious diseases. When qualifying an antibody, or an antigen-binding fragment thereof, means that said antibody, or antigen-binding fragment thereof, specifically binds to or specifically recognizes a pathogen microorganism.

[0040] “Antibody (Ab)” and “immunoglobulin (Ig)” may be used interchangeably and refer to a protein having a combination of two heavy chains (H chains) and two light chains (L chains). In particular, the term “antibodies” refers to such assemblies which have significant known specific immunoreactive activity to an antigen of interest (e.g., one of infectious agents such as bacteria). As mentioned above, antibodies and immunoglobulins comprise light and heavy chains, with or without an interchain covalent linkage between them. Basic immunoglobulin structures in vertebrate systems are relatively well-understood. The generic term “immunoglobulin” comprises five distinct classes of immunoglobulins that can be distinguished biochemically: IgG, IgM, IgA, IgD, and IgE. IgG immunoglobulins comprise two identical light chains with a molecular weight of about 23 kDa, and two identical heavy chains with a molecular weight of about 53-70 kDa. The four chains are joined by disulfide bonds in a “ Y” configuration wherein the light chains bracket the heavy chains starting at the mouth of the “Y” and continuing through the variable region. The light chains of an immunoglobulin are classified as either kappa (κ) or lambda (λ). Each heavy chain class may be bonded with either a κ or λ light chain. In general, the light and heavy chains are covalently bonded to each other, and the “tail” regions of the two heavy chains are bonded to each other by covalent disulfide linkages or non-covalent linkages when the immunoglobulins are generated either by hybridomas, B cells or genetically engineered host cells. In the heavy chains, the amino acid sequence runs from the N-terminus at the forked ends of the Y configuration to the C-terminus at the bottom of each chain. Those skilled in the art willappreciate that heavy chains are classified as gamma (y), mu (p), alpha (a), delta (6), or epsilon (e) with some subclasses among them (e.g., yl-y4). It is the nature of the heavy chain that determines the “class” of the antibody as IgG, IgM, IgA, IgD, or IgE, respectively. The immunoglobulin subclasses or “isotypes” (e.g., IgGl, IgG2, IgG3, IgG4, IgAl, etc.) are well- characterized and are known to confer functional specialization. The variable region of an antibody allows the antibody to selectively recognize and specifically bind to an epitope on an antigen. That is, the light chain variable region (VL) and heavy chain variable region (VH) of an antibody combine to form the variable region that defines a three-dimensional antigen¬ binding site. This quaternary antibody structure thus forms the antigen-binding site present at the end of each arm of the “Y”. More specifically, the antigen-binding site is defined by three complementarity determining regions (CDRs) on each of the VH and VL.

[0041] “Antigen-binding fragment” of an antibody, which is interchangeable with the term “antigen-binding domain” of an antibody, refers to a part or region of an antibody, or immunoglobulin, which comprises fewer amino acid residues than a whole antibody, or immunoglobulin, and which is capable of binding to an antigen and / or of competing with a whole antibody for antigen binding (e.g., for specific binding to a bacterial antigen). Examples of antigen-binding antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv fragments, scFv fragments, disulfide-linked Fvs (sdFv), a Fd fragment consisting of the VH and CHI domains, linear antibodies, single domain antibodies such as sdAb (either VL or VH), camelid VHH domains, multi-specific antibodies formed from antibody fragments such as a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region, and an isolated CDR of an antibody. Papain digestion of antibodies produces two identical antigen-binding fragments, called “Fab” fragments, and a residual “Fc” fragment, a designation reflecting the ability to crystallize readily. A Fab fragment consists of one entire L chain (variable region of the L chain (VL) and constant domain of the L chain (CL)), along with part of one H chain consisting of the variable region of the H chain (VH) and the first constant domain of the heavy chain (CHI). Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a single antigen-binding site. Pepsin treatment of an antibody yields a single large F(ab')2 fragment that roughly corresponds to two disulfide-linked Fab fragments having divalent antigen-binding activity and is still capable of crosslinking antigen. Fab' fragments differ from Fab fragments by having additional few amino acid residues at the carboxy terminus of the CHI domain including one or more cysteines from the antibody hingeregion. Fab'-SH is the designation for Fab' in which the cysteine residue(s) of the constant domains bear(s) a free thiol group. F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments that have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0042] “Antigen” or “Ag” as used herein refers to a protein (e.g., a bacterial protein), which is specifically recognized by an antibody or antibody -binding fragment thereof.

[0043] “CDR” or “complementarity determining region” means the non-contiguous antigen combining sites found within the variable region of both the heavy chain and the light chain. The precise amino acid sequence boundaries of a given CDR can be determined using any of a number of well-known schemes, including those described by Kabat et al. (1991), “Sequences of Proteins of Immunological Interest” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (“Kabat” numbering scheme); Al-Lazikani et al., (1997) JMB 273,927-948 (“Chothia” numbering scheme); or a combination thereof such as the AbM definition which is a compromise between the two used by Oxford Molecular's AbM antibody modelling software. More recently, a universal numbering system has been developed and widely adopted, ImMunoGeneTics (IMGT) Information System® (Lefranc et al., Nucleic Acids Res. 27: 209-212 1999). IMGT® is an integrated information system specializing in immunoglobulins (Ig), T cell receptors (TCR) and major histocompatibility complex (MHC) of human and other vertebrates. Herein, the CDRs are referred to in terms of both the amino acid sequence and the location within the light or heavy chain e.g., VH-CDR1, VH-CDR2, VH- CDR3, VL-CDR1, VL-CDR2, VL-CDR3). AS the "location" of the CDRs within the structure of the immunoglobulin variable region is conserved between species and present in structures called loops, by using numbering systems that align variable region sequences according to structural features, CDR and framework amino acid residues may be readily identified. This information can be used in grafting and replacement of CDRs from immunoglobulins of one species into an acceptor framework from, typically, a human antibody. Correspondence between the Kabat numbering and the IMGT® unique numbering system is also well-known to one skilled in the art (e.g., Lefranc et al., supra).

[0044] “Comprise” or a variant thereof (e.g., “comprises”, “comprising”) is used herein according to common patent application drafting terminology. Hence, “comprise” preceded byan object and followed by a constituent means that the presence of a constituent in the object is required (typically as a component of a composition), but without excluding the presence of any further constituent(s) in the object. Moreover, any occurrence of “comprise” or a variant thereof herein also encompasses narrower expression “substantially consist of’, further narrower expression “consist of’ and any variants thereof (e.g., “consists of’, “consisting of’), unless otherwise stated.

[0045] “Epitope” refers to a specific arrangement of amino acids located on a protein or proteins to which an antibody, or an antigen-binding fragment thereof, specifically binds. Epitopes often consist of a chemically active surface grouping of molecules such as amino acids or sugar side chains, and have specific three-dimensional structural characteristics as well as specific charge characteristics. Epitopes can be linear (or sequential) or conformational, i.e., involving two or more sequences of amino acids in various regions of the antigen that may not necessarily be contiguous.

[0046] “Fc domain,” “Fc portion,” and “Fc region” may be used interchangeably and refer to a C-terminal fragment of an antibody heavy chain, e.g., from about amino acid (aa) 230 to about aa 450 of human gamma heavy chain or its counterpart sequence in other types of antibody heavy chains (e.g., a, 5, 8 and p for human antibodies), or a naturally occurring allotype thereof.

[0047] “Framework region” or “FR region” or “non-CDR region” includes the amino acid residues that are part of the variable region, but are not part of the CDRs (e.g., using the Kabat definition of CDRs, the Chothia definition of CDRs, or the IMGT® numbering definition of CDRs). Therefore, a variable region framework ranges from about 100 to 120 amino acids in length but includes only those amino acids outside of the CDRs.For the specific example of a heavy chain variable region (VH) and for the CDRs as defined by Kabat or Chothia:- FR1 may correspond to the domain of the variable region encompassing amino acids 1- 25 according to Chothia / AbM's definition, or 5 amino acid residues later according to Kabat's definition;- FR2 may correspond to the domain of the variable region encompassing amino acids 36-49;- FR3 may correspond to the domain of the variable region encompassing amino acids 67-98; and- FR4 may correspond to the domain of the variable region from amino acid 104-110 to the end of the variable region.The framework regions for the light chain are similarly separated by each of the CDRs of the light chain variable region (VL).In naturally occurring antibodies, the six CDRs present on each monomeric antibody are short, non-contiguous sequences of amino acids that are specifically positioned to form the antigen¬ binding site as the antibody assumes its three-dimensional configuration in an aqueous environment. As indicated above, the remainders of the heavy and light variable regions show less inter-molecular variability in amino acid sequence and correspond to the framework regions. The framework regions largely adopt a P-sheet conformation and the CDRs form loops which connect, and in some cases form part of, the P-sheet structure. Thus, these framework regions act to form a scaffold for positioning the six CDRs in correct orientation by inter-chain, non-covalent interactions. The antigen-binding site formed by the positioned CDRs defines a surface complementary to the epitope on the immunoreactive antigen. This complementary surface promotes the non-covalent binding of the antibody to the immunoreactive antigen epitope.

[0048] “Heavy chain region” includes amino acid sequences derived from the constant domains of an immunoglobulin heavy chain. A protein comprising a heavy chain region comprises at least one of a CHI domain, a hinge region (e.g., upper, middle, and / or lower hinge domain), a CH2 domain, a CH3 domain, or a variant or fragment thereof. In some embodiments, the antibody, or the antigen-binding fragment thereof, as described herein may comprise the Fc region of an immunoglobulin heavy chain (e.g., a hinge portion, a CH2 domain, and a CH3 domain). In some embodiments, the antibody, or the antigen-binding fragment thereof, as described herein lacks at least a region of a constant domain (e.g., all or part of a CH2 domain). In some embodiments, at least one, and preferably all, of the constant domains are derived from a human immunoglobulin heavy chain. For example, in some embodiments, the heavy chain region comprises a fully human hinge domain. In some embodiments, the heavy chain region comprises a fully human Fc region (e.g., hinge, CH2 andCH3 domain from a human immunoglobulin). In some embodiments, the constituent constant domains of the heavy chain region are from different immunoglobulin molecules. For example, a heavy chain region of a protein may comprise a CH2 domain derived from an IgGl molecule and a hinge region derived from an IgG3 or IgG4 molecule. In some embodiments, the constant domains are chimeric domains comprising regions of different immunoglobulin molecules. For example, a hinge may comprise a first region from an IgGl molecule and a second region from an IgG3 or IgG4 molecule. In some embodiments, the constant domains of the heavy chain region may be modified such that they vary in amino acid sequence from the naturally occurring (wild-type) immunoglobulin molecule. That is, the antibody, or the antigen-binding fragment thereof, as described herein may comprise alterations or modifications to one or more of the heavy chain constant domains (CHI, hinge, CH2 or CH3) and / or to the light chain constant domain (CL). Exemplary modifications include additions, deletions or substitutions of one or more amino acids in one or more domains.

[0049] “Hinge region” includes the region of a heavy chain molecule that joins the CHI domain to the CH2 domain. This hinge region comprises approximately 25 amino acid residues and is flexible, thus allowing the two N-terminal antigen-binding regions to move independently. Hinge regions can be subdivided into three distinct domains: upper, middle, and lower hinge domains.

[0050] “Infection” refers to any undesired presence and / or growth of pathogen (typically bacteria, viruses, fungi, mycobacteria or parasites) in a subject. Such undesired presence of microorganism may have a negative effect on the host subject's health and well-being. While the term “infection” should not be taken as encompassing the normal growth and / or presence of microorganism which are normally present in the subject, for example in the digestive tract of the subject, it may encompass the pathological overgrowth of such microorganism. Infections may be caused by the growth and / or presence of microorganism, typically bacteria, viruses, fungi, mycobacteria or parasites.

[0051] “Identity” or “identical”, when used in the present invention in a relationship between the sequences of two or more polypeptides or of two or more nucleic acids, refers to the degree of sequence relatedness between polypeptides or nucleic acids (respectively), as determined by the number of matches between strings of two or more amino acid residues or of two or morenucleotides, respectively. “Identity” measures the percent of identical matches between the smaller of two or more sequences with gap alignments (if any) addressed by a particular mathematical model or computer program (z.e., “algorithms”). Identity of related polypeptides or nucleic acid sequences can be readily calculated by known methods. Such methods include, but are not limited to, those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part 1, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M. Stockton Press, New York, 1991; and Carillo et al., SIAM J. Applied Math. 48, 1073 (1988). Preferred methods for determining identity are designed to give the largest match between the sequences tested. Methods of determining identity are described in publicly available computer programs. Preferred computer program methods for determining identity between two sequences include the GCG program package, including GAP (Devereux et al., Nucleic Acids Res. 1984 Jan 11; 12(1 Pt l):387-95; Genetics Computer Group, University of Wisconsin, Madison, Wis.), BLASTP, BLASTN, and FASTA (Altschul etal., J. Mol. Biol. 215, 403-410 (1990)). The BLASTX program is publicly available from the National Center for Biotechnology Information (NCBI) and other sources (BLAST Manual, Altschul et al. NCB / NLM / NIH Bethesda, Md. 20894; Altschul et al., J. Mol. Biol. 215, 403-410 (1990)). The well-known Smith Waterman algorithm may also be used to determine identity.

[0052] " Isolated" or "non-naturally occurring" with reference to a biological component (such as an antibody or a nucleic acid) refers to a biological component altered or removed from the natural state. For example, an antibody or a nucleic acid naturally present in a living animal is not "isolated" but the same antibody or nucleic acid partially or completely separated from the coexisting materials of its natural state is “isolated”. An isolated antibody or nucleic acid can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell. Typically, a preparation of isolated antibody or nucleic acid contains the antibody or nucleic acid at least about 80% pure, at least about 85% pure, at least about 90% pure, at least about 95% pure, greater than 95% pure, greater than about 96% pure, greater than about 97% pure, greater than about 98% pure, or greater than about 99% pure. Nucleic acids and proteins, such an antibodies, that are "non-naturally occurring" or have been"isolated" include nucleic acids and proteins purified by standard purification methods. The term also embraces nucleic acids and proteins prepared by recombinant expression in a host cell as well as chemically synthesized nucleic acids.

[0053] “Human” refers to a subject of both genders and at any stage of development (z'.e., neonate, infant, juvenile, adolescent, adult).

[0054] “Infectious disease” refers to a pathologic condition or disorder resulting from an infection. Examples of specific infections include “bacterial disease”, “viral disease”, “fungal disease”, “mycobacterial disease” and “parasitic disease”, which are infectious diseases caused respectively by bacteria, viruses, fungi, mycobacteria or parasites. Therapeutic agents for the treatment of infectious diseases are “anti-infective” agents.

[0055] “Monoclonal antibody (mAb)” refers to an antibody obtained from a population of substantially homogeneous antibodies, z.e., the individual antibodies comprised in the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to polyclonal antibody preparations that include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant (epitope) on the antigen. In addition to their specificity, the monoclonal antibodies are advantageous in that they may be synthesized uncontaminated by other antibodies. The modifier “monoclonal” is not to be construed as requiring production of the antibody by any particular method. For example, a monoclonal antibody, or an antigen-binding fragment thereof, as described herein may be prepared by the well-known hybridoma methodology, or may be made using recombinant DNA methods in bacterial, eukaryotic, animal, or plant cells. The “monoclonal antibodies” may also be isolated from phage antibody libraries using techniques commonly known in the field.

[0056] As used herein, the term “nucleic acid” or “polynucleotide” refers to a polymer of nucleotides covalently linked by phosphodiester bonds, such as deoxyribonucleic acids (DNA) or ribonucleic acids (RNA), in either single- or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particularnucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, SNPs, and complementary sequences as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues.

[0057] “Patient” refers to a warm-blooded animal, more preferably a human, who / which is awaiting the receipt of, or is receiving medical care or is / will be the object of a medical procedure.

[0058] “Pharmaceutically acceptable” means that the ingredients of a composition are compatible with each other and not deleterious to the patient to which / whom it is administered.

[0059] “Pharmaceutically acceptable carrier” refers to an excipient that does not produce an adverse, allergic or other untoward reaction when administered to an animal, preferably a human. It includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. For human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by regulatory offices, such as, e.g., FDA Office or EMA.

[0060] “Pharmaceutical composition” refers to a composition comprising at least one therapeutic agent and at least one pharmaceutically acceptable carrier.

[0061] “Selected from” is used herein according to common patent application drafting terminology, to introduce a list of elements among which one or more item(s) is (are) selected.

[0062] “Single-chain Fv”, also abbreviated as “sFv” or “scFv”, refers to a fusion protein comprising at least one antibody fragment comprising a variable region of a light chain (VL) and at least one antibody fragment comprising a variable region of a heavy chain (VH), wherein the light and heavy chain variable regions are contiguously linked, e.g., via a synthetic linker, such as a short flexible polypeptide linker, and capable of being expressed as a single chain polypeptide, and wherein the scFv retains the specificity of the intact antibody from which it is derived.

[0063] “Subject” refers to an animal, typically a warm-blooded animal, preferably a mammal, more preferably a primate, furthermore preferably a human. In one embodiment, the subject is a “patient” as defined herein. In one embodiment, the subject is affected, preferably is diagnosed, with a disease. In one embodiment, the subject is at risk of developing a disease. Examples of risks factor include, but are not limited to, genetic predisposition, or familial history of the disease (e.g., infectious agents). In some embodiments, the subject is an adult (for example a subject above the age of 18). In some embodiments, the subject is a child (for example a subject below the age of 18). In some embodiments, the subject is a male. In some embodiments, the subject is a female.

[0064] “Therapeutic agent”, “active pharmaceutical ingredient” and “active ingredient” refer to a compound for therapeutic use and relating to health. Especially, a therapeutic agent may be indicated for treating a disease. An active ingredient may also be indicated for improving the therapeutic activity of another therapeutic agent.

[0065] “Therapeutically effective amount” (in short “effective amount”) refers to the amount of a therapeutic agent that is sufficient to achieve the desired therapeutic, prophylactic or preventative effect in the patient to which / whom it is administered, without causing significant negative or adverse side effects to said patient. A therapeutically effective amount may be administered prior to the onset of the disease, disorder, or condition, for a prophylactic or preventive action. Alternatively, or additionally, the therapeutically effective amount may be administered after initiation of the disease, disorder, or condition, for a therapeutic action.

[0066] “Treating”, “treatment” or “alleviation” refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) the targeted pathologic condition or disorder (z.e., a “disease”). Those in need of treatment include those already with the disease as well as those prone to have the disease or those in whom the condition or disease is to be prevented. A patient is successfully “treated” for a disease if, after receiving a therapeutic amount of a therapeutic agent, the patient shows observable and / or measurable reduction in or absence of one or more of the following: reduction in the number of pathogens e.g., infectious agents); reduction in the percent of total cells that are pathogenic; and / or relief to some extent, one or more of the symptoms associated with the specific disease; reduced morbidity and mortality, and improvement in quality of lifeissues. The above parameters for assessing successful treatment and improvement in the disease are readily measurable by routine procedures familiar to a physician.

[0067] “Variable”, “variable region” or “variable domain” refer to the fact that certain regions of the variable domains VH and VL differ extensively in sequence among antibodies and are used in the binding and specificity of each particular antibody for its target antigen. However, the variability is not evenly distributed throughout the variable domains of antibodies. It is concentrated in three segments called “hypervariable loops” in each of the VL domain and the VH domain which form part of the antigen-binding site. The 6 hypervariable loops may each comprise part of a CDR, as defined hereinabove.DETAILED DESCRIPTIONMolecular systemGeneral structure

[0068] This invention relates to a molecular system as defined herein, which may alternatively be referred to as a “targeted dynamic antimicrobial system”, a “drug delivery system”, an “anti-infective agent delivery system”, or a “non-covalent ordered system”.

[0069] According to the invention, the molecular system comprises:(i) at least one dynamic constitutional framework (DCF) comprising: a plurality of cores and a plurality of connectors, wherein each of the connectors is covalently bound in a reversible way to at least two of the cores;(ii) a plurality of functional heads, wherein each of the functional heads is covalently bound in a reversible way to one of the cores;(iii) at least one pillararene; and(iv) at least one anti-infective agent; wherein each of the anti-infective agents is hosted in one of the pillararenes; and wherein each of the pillararenes is covalently bound in a reversible way to at least one of the cores.

[0070] In other words, the molecular system of the present invention can be described as a dynamic constitutional framework (DCF) that is both functionalised by functional heads as defined herein and drug-loaded by at least one pillararene hosting at least one anti-infective agent (“host-guest complex”).

[0071] According to some preferred embodiments, the molecular system is substantially as described in any one of the Examples hereinbelow. In some preferred embodiments, the molecular system is selected from DCF-C-APAl-Lev (Bis(3-aminopropyl)PEG-BTA polymer grafted with aminoguanidine and Deca(6-aminohexyl-l-oxy)pillar[5]arene 0 Levofloxacin complex) and DCF-C-APA2-Lev (Bis(3-aminopropyl)PEG-BTA polymer grafted with aminoguanidine and Deca((l-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-lH-l,2,3-triazol-4-yl)methoxy)pillar[5]arene 0 Levofloxacin complex), as defined in any one of the Examples hereinbelow. In some preferred embodiments, the molecular system is selected from DCF-C-AHPA2-Lev (Bis(3-aminopropyl)PEG-BTA polymer grafted with aminoguanidine and Deca(Pentanehydrizide-lH-l,2,3-triazol-4-yl)methoxy)pillar[5]arene 0 Levofloxacin complex). In some preferred embodiments, the molecular system is selected from DCF-C- APAl-Lev, DCF-C-APA2-Lev, and DCF-C-AHPA2-LevDynamic constitutional framework (DCF)

[0072] The dynamic constitutional framework (DCF) as defined herein may alternatively be referred to as a “polymeric network”, a “3D network”, a “3D polymeric network”, or a “3D polymeric-dynamic network”. For the sake of conciseness and readability, “dynamic constitutional framework”, “DCF” and the like are used herein irrespective of whether the DCF is incorporated in the molecular system, (e.g., irrespective of whether it is functionalised and / or drug-loaded), which is in line with the terminology generally used in the art. However, “DCF” without further determination typically refers to the DCF when incorporated in the molecular system.

[0073] In the present invention, each core is independently selected from phenyl, monocyclic heteroaryl (e.g., pyridinyl), polycyclic aryl (e.g., naphthyl or pyrenyl), and polycyclic heteroaryl (e.g, bipyridinyl or indolyl); wherein each of the cores is covalently bound to at least three groups selected from the connectors, the functional heads, and the pillararenes (z'.e., each core is at least trivalent). According to some preferred embodiments, each of the cores iscovalently bound to exactly three groups selected from the connectors, the functional heads, and the pillararenes (z.e., each core is trivalent).

[0074] The cores bond to at least three groups selected from the connectors, the functional heads, and the pillararenes (“other groups”) forms the base network of the molecular system, and therefore are mentioned herein as a feature of the present invention. However, because of the network nature of the molecular system, some cores in the molecular system will necessarily be bound to only one or two other group(s). Consequently, the molecular system according to the invention is not limited to a molecular system including only cores bound to three other groups. The amount of cores bound to only one or two group(s) in a given molecular system depends on how the molecular system is prepared and, in particular, on the initial amounts of core and other group(s) at each step of the manufacturing process.

[0075] According to some embodiments, each core is independently selected from trivalent phenyl and trivalent monocyclic heteroaryl (e.g., pyridinyl). In some preferred embodiments, each core is a trivalent phenyl. In some further preferred embodiments, each core is a 1,3,5-subsituted phenyl.

[0076] In the present invention, each connector is a bifunctional linear homopolymer. “Bifunctional linear homopolymer” refers herein to a polymer that consists of one kind of structural unit, and all structural units exist in a single line with no branches or intramolecular bridges; and the terminated groups are functionalized with desired functional groups. According to some preferred embodiments, the two functions are coupling functions or a (C1-C4) alkyl terminated by at least one coupling function.

[0077] According to some embodiments, each connector is independently selected from water-soluble bifunctional linear homopolymer such as bifunctional linear polyethylene glycol (PEG) and bifunctional linear polypropylene glycol (PPG).

[0078] According to some embodiments, each connector is independently selected from polyethylene glycol (PEG) terminated by two (C1-C4) alkyls and polypropylene glycol (PPG) terminated by two (C1-C4) alkyls. According to some preferred embodiments, each connector is a polyethylene glycol (PEG) terminated by two propyls. In some preferred embodiments, each connector is a polyethylene glycol (PEG) terminated by two n-propyls.

[0079] In the present invention, each of the connectors is covalently bound to two of the cores (z'.e., “to exactly two of the cores”) by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product. Thereby, each of the connectors is covalently bound in a reversible way to at least two of the cores. In some embodiments, each of the connectors is covalently bound to at least two of the cores by means of a reversible coupling product. In some preferred embodiments, the reversible coupling product is N=CH (z.e., the bond is an imine bond).

[0080] The connectors bond to two of the cores form the base network of the DCF, and therefore are mentioned herein as a feature of the present invention. However, because of the network nature of the DCF, some connectors in the molecular system will necessarily be bound to only one core, instead of two. Consequently, the molecular system according to the invention is not limited to a DCF including only connectors bound to two cores. The amount of connectors bound to only one core in a given DCF depends on how the DCF is prepared and, in particular, on the initial amounts of core and connectors.

[0081] According to some embodiments, a DCF not functionalised by functional heads (herein “base DCF”) is obtained by reaction of a precursor of a core (herein “pre-core”) with at least one precursor of a connector (herein “pre-connector”); preferably the reaction between at least one coupling function of the pre-core and at least one coupling function of the pre-connector(s); more preferably the reaction between exactly one coupling function of the pre-core and exactly one coupling function of the pre-connector(s) (thereby, each core is bound only once to a given connector, whereas each connector is bound to one or two different cores). In some embodiments, the pre-core comprises at least three coupling functions (e.g., aldehyde), preferably exactly three coupling functions. In some embodiments, the pre-connector comprises at least two coupling functions (e.g., NH2), preferably exactly two coupling functions. In some preferred embodiments, the pre-core comprises three coupling functions and the pre-connector comprises at least two coupling functions.

[0082] In some embodiments, the initial moral ratio between the pre-core and the pre-connector ranges from about 0.5 to 2, preferably the initial moral ratio is about 1.

[0083] In some preferred embodiments, the pre-core is 1,3,5-benzenetrialdehyde (BTA). In some preferred embodiments, the pre-connector is bi s(3 -aminopropyl) terminated polyethyleneglycol (PEG) (i.e., H2N-CH2CH2CH2-(OCH2CH2)n-O-CH2CH2CH2-NH2; wherein, typically, n ranges from 29 to 40 and / or the molecular weight of the PEG ranges from 1 400 to 1 900 Da). In some further preferred embodiments, the base DCF is obtained by reaction of 1,3,5-benzenetrialdehyde (BTA) with bi s(3 -aminopropyl) terminated polyethylene glycol (PEG).Functional head

[0084] In the present invention, each of the functional heads is covalently bound to one of the cores (i.e., “to exactly one of the cores”, in other words, no functional head can be bound to two or more cores) by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product. In some embodiments, each of the functional heads is covalently bound to one of the cores by means of a reversible coupling product. In some preferred embodiments, the reversible coupling product is N=CH i.e., the bond is an imine bond). Thereby, each of the functional heads is covalently bound in a reversible way to one of the cores.

[0085] In the present invention, each functional head is independently selected from amines, polyamines, hydrazides, and acyl-hydrazides. According to some embodiments, the function head comprises at least one amino (-NH2) group.

[0086] According to some embodiments, each functional head is independently selected from the following formulae (A’) to (W’)wherein one (z.e., “exactly one”) of the -NH2 groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the functional head is covalently bound to one of the cores.

[0087] In some preferred embodiments, each functional head is independently selected from the following formulae (B’) and (C’)NH2HN NH (C')NH2wherein one (i.e., “exactly one”) of the -NH2 groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the functional head is covalently bound to one of the cores; and wherein Mnis 2000 Da in the compound of formula (B’).Functionalised-DCF

[0088] To prepare the molecular system, this invention can use a DCF functionalised by the functional heads (herein “functionalised-DCF”), wherein each of the functional heads is covalently bound to one of the cores.

[0089] According to some embodiments, the functionalised-DCF is obtained by reaction of a base DCF as described herein with a precursor of the functional head (herein “pre-(functional head)”); preferably by the reaction between at least one coupling function of the base DCF (e.g., aldehyde) and at least one coupling function of the pre-(functional head) (e.g., NH2); more preferably between exactly one coupling function of the base DCF and exactly one coupling function of the pre-(functional head).

[0090] In some embodiments, the initial moral ratio between the base DCF and the pre-(functional head) ranges from about 0.5 to 1.5, preferably the initial moral ratio is about 1.

[0091] According to some embodiments, each pre-(functional head) is independently selected from the following formulae (A) to (W).

[0092] In some preferred embodiments, each pre-(functional head) is independently selected from the following formulae (B) and (C)NH2HN^NH (C)I NH2wherein Mnis 2000 Da in the compound of formula (B).Pillararene

[0093] For the sake of conciseness and readability, “pillararene” and the like are used herein irrespective of whether the pillararene is incorporated in the molecular system, which is in line with the terminology generally used in the art. However, “pillararene” without further determination typically refers to the pillararene when incorporated in the molecular system.

[0094] In the present invention, each of the pillararenes is covalently bound to at least one of the cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product. In some embodiments, each of the pillararenes is covalently bound to at least one of the cores by means of a reversible coupling product. In some preferred embodiments, the reversible coupling product is N=CH (z.e., the bond is an imine bond). Thereby, each of the pillararenes is covalently bound in a reversible way to at least one of the cores. In some embodiments, each of the pillararenes is covalently bound to one of the cores (z.e., “to exactly one of the cores”, in other words, no pillararene can be bound to two or more cores).

[0095] According to some preferred embodiments, the pillararene is a pillar[5]arene. In some embodiments, the pillar[5]arene is a compound of formula (P)whereineach L1independently represents -O-alkyl, -O-polyether or -O-polyamine;the oxygen atom is bond to the phenyl to which L1is bound, andthe alkyl, polyether or polyamine is optionally interrupted or terminated by at least one coupling product (e.g., triazolyl), preferably by exactly one coupling product; andeach R1independently represents -NH2, -NH3+, or a reversible coupling product (e.g., N=CH).

[0096] In some embodiments, each L1represents the same group as the others. In some embodiments, each R1represents the same group as the others, except for one R1that represents a coupling product. In some embodiments, each L1represents -O-alkyl. In some embodiments, each L1represents -O-polyether terminated by one coupling product. In some embodiments, each coupling product terminating the -O-alkyl is triazolyl.

[0097] In some preferred embodiments, the pillar[5]arene is selected from the following formulae (APAl’) and (APA2’)(APAl')(APA2')NH2wherein at least one of the -NH2 or -NH3+groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the pillararene is covalently bound to at least one of the cores.

[0098] In some preferred embodiments, the pillar[5]arene is the compound of formula (AHPA2’)(AHPA2’)wherein at least one of the -NH2 or -NH3+groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the pillararene is covalently bound to at least one of the cores.

[0099] In some preferred embodiments, exactly one among the -NH2 or -NH3+groups is replaced by either -NH- or a reversible coupling product.DCF-pillararene system

[0100] This invention also relates to a functionalised-DCF as defined herein, which further comprises at least one pillararene as defined herein (herein “DCF-pillararene system” or “hosting system”), wherein each of the pillararenes is covalently bound to at least one of the cores, and each of the pillararenes does not host any anti-infective agent. According to some embodiments, each of the pillararenes does not host any therapeutic agent. This DCF-pillararene system is useful, for example, as synthetic intermediate for preparing the molecular system comprising an anti -infective agent hosted in the pillararenes, as described herein.

[0101] Therefore, this invention also relates to a hosting system comprising:(i) at least one dynamic constitutional framework (DCF) comprising a plurality of cores and a plurality of connectors, as defined herein;(ii) a plurality of functional heads as defined herein;wherein each of the functional heads is covalently bound to one of the cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product;(iii) at least one pillararene;wherein each of the pillararenes is covalently bound to at least one of the cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product;wherein each of the cores is covalently bound to at least three groups selected from the connectors, the functional heads, and the pillararenes andwherein each of the pillararenes does not host any anti-infective agent.

[0102] According to some embodiments, the DCF-pillararene system is obtained by reaction of a functionalised-DCF as defined herein with at least one pillararene (or “precursor of a pillararene”); preferably the reaction between at least one coupling function of the functionalised-DCF (e.g., aldehyde) and at least one coupling function of the pillararene (e.g., NH2); more preferably the reaction between exactly one coupling function of the functionalised-DCF and exactly one coupling function of the pillararene (thereby, each core of the functionalised-DCF is bound only once to a given pillararene).

[0103] In some embodiments, the initial moral ratio between the functionalised-DCF and the pillararene(s) ranges from about 0.0033 to 0.82, preferably the initial moral ratio is about 0.05. In some embodiments, the initial equivalent moral ratio between the functionalised-DCF and the pillararene(s) ranges from about 0.033 to 8.2, preferably the initial equivalent moral ratio is about 0.5. By “equivalent moral ratio” it is meant the molar ratio taking into account the number of free functions (i.e., the functions available for reaction) in the reactants. For example, when the functionalised-DCF has one free aldehyde group and the pillararene has tenfree amine groups, an initial molar ratio of about 0.05 corresponds to an initial equivalent molar ratio of about 0.5.

[0104] In some preferred embodiments, the pillar[5]arene (or “precursor of a pillar[5]arene”) is selected from the following formulae (APA1) and (APA2).(APA1)(APA2)NH3

[0105] In some preferred embodiments, the (or “precursor of a pillar[5]arene”) is the compound of formula (AHPA2)(AHPA2)wherein at least one of the -NH2 or -NH3+groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the pillararene is covalently bound to at least one of the cores.Anti-infective agent

[0106] According to some embodiments, the anti-infective agent is an antibiotic agent. In some embodiments, the anti -infective agent is an antibiotic agent selected from aminoglycosides, cephalosporins, fluoroquinolones, lincosamides, macrolides, penicillins, sulphonamides, tetracyclines, and other antibiotics known in the art such as daptomycin, metronidazole, telithromycin, vancomycin, teicoplanin, and the like.

[0107] In some embodiments, the anti-infective agent is an antibiotic agent selected from: - Aminoglycosides such as, for example, amikacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, streptomycin, or tobramycin;- Cephalosporins such as, for example, cefaclor, cefadroxil, cefalexin (Cef), cefamandole, cefazolin, cefdinir, cefepime, cefonicid, cefoperazone, cefotaxime, cefotetan, cefoxitin, cefpodoxime, cefprozil, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefuroxime, cephalexin, cephapirin, cephradine, or cefditoren;- Fluoroquinolones such as, for example, ciprofloxacin, enoxacin, gatifloxacin, gemifloxacin, levofloxacin (Lev), lomefloxacin, moxifloxacin, norfloxacin, ofloxacin, sparfloxacin, or trovafloxacin;- Lincosamides such as, for example, clindamycin, lincomycin (Lin), or pirlimycin;- Macrolides such as, for example, azithromycin, erythromycin, clarithromycin, dirithromycin, roxithromycin, or telithromycin;- Penicillins such as, for example, amoxicillin, ampicillin, bacampicillin, carbenicillin, cioxacillin, dicloxacillin, flucioxacillin, mezlocillin, nafcillin, oxacillin, penicillin G, penicillin V, piperacillin pivampicillin, pivmecillinam, or ticarcillin;- Sulphonamides such as, for example, sulfamethizole, sulfamethoxazole, sulfisoxazole, trimethoprim, or sulfamethoxazole;- Tetracyclines such as, for example, demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline, ortigecycline; and-other antibiotics known in the art such as, for example, daptomycin, metronidazole, telithromycin, vancomycin, teicoplanin, and the like.

[0108] According to some preferred embodiments, the antibiotic agent is selected from fluoroquinolones, lincosamides and cephalosporins. In some preferred embodiments, the antibiotic agent is selected from levofloxacin (Lev), lincomycin (Lin) and cefalexin (Cef). In some further preferred embodiments, the antibiotic agent is levofloxacin (Lev).Host-guest complex

[0109] In the present invention, each anti-infective agent is hosted in one of the pillararene(s) (z.e., “in exactly one of the pillararenes”, in other words, no anti -infective agent can be hosted in two or more pillararenes), thereby forming an “host-guest complex”. “Hosted” refers to the establishment of non-covalent interactions with dynamic equilibrium between the unbound and the bound states, so that the association of the guest and the host is stable enough to allow transport processes, but weak enough to allow a delivery of the guest in the desired biological target (cell, organ, biofilm, etc.).

[0110] According to some embodiments, the host-guest complex is obtained by contacting and / or reacting the pillararene with the anti -infective agent(s). According to some preferred embodiments, the host-guest complex is obtained by contacting the pillararene with the anti-infective agent(s). In some embodiments, the initial moral ratio between the pillararene and the total amount of anti-infective agent(s) ranges from about 0.014 to 124, preferably from about 0.14 to 12.4, more preferably the initial moral ratio is about 2.

[0111] In the present invention, each of the host-guest complexes is covalently bound to at least one of the cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product. In some embodiments, each of the host-guest complexes is covalently bound to at least one of the cores by means of a reversible coupling product. In some preferred embodiments, the reversible coupling product is N=CH (z.e., the bond is an imine bond). Thereby, each of the host-guest complexes is covalently bound in a reversible way to at least one of the cores. In some embodiments, each of the host-guest complexes is covalently bound to one of the cores (z.e., “to exactly one of the cores”, in other words, no host-guest complex can be bound to two or more cores).Antibody

[0112] According to some embodiments, the molecular system further comprises at least one antibody.

[0113] In some embodiments, the antibody specifically binds to a cell surface of bacteria, preferably bacteria of the genus Acinetobacter, more preferably bacteria of the species Acinetobacter baumannii. In some embodiments, the antibody specifically binds to a protein from bacteria, preferably bacteria of the genus Acinetobacter, more preferably bacteria of the species Acinetobacter baumannii. In some embodiments, the antibody specifically binds to a surface protein from bacteria, preferably bacteria of the genus Acinetobacter, more preferably bacteria of the species Acinetobacter baumannii. In some embodiments, the antibody specifically binds to a surface from virulent bacteria, preferably a capsulated bacterial strain, more preferably, virulent bacteria of the genus Acinetobacter, more preferably bacteria of the species Acinetobacter baumannii.

[0114] In some embodiments, the antibody specifically binds to the protein Omp25 from Acinetobacter baumannii. Typically, Omp25 from Acinetobacter baumannii has an amino acid sequence having at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% of amino acid sequence identity with SEQ ID NO: 4.

[0115] In some embodiments, the antibody, or antigen-binding fragment thereof, may be polyclonal or monoclonal. Preferably, the antibody, or antigen-binding fragment thereof, is monoclonal.

[0116] In some embodiments, the antibody may be a whole antibody or an antigen-binding fragment of an antibody, preferably an antigen-binding fragment of an antibody.

[0117] In some embodiments, the antigen-binding fragment is a molecule selected from the group comprising or consisting of an immunoglobulin single variable domain (ISVD), a single domain antibody (sdAb), a Variable Heavy domain of Heavy chain (VHH), a nanobody, a single-chain antibody or scFv, a dimeric single chain antibody or di-scFv, a single-domain antibody, aFv, a Fab, a Fab', aFab'-SH, aF(ab)’2, aFd, a defucosylated antibody, abi-specific antibody, a diabody, a triabody and a tetrabody. In some embodiments, the antigen-binding fragment is a single-chain antibody selected from the group comprising or consisting of an immunoglobulin single variable domain (ISVD), a single domain antibody (sdAb), a Variable Heavy domain of Heavy chain (VHH), a nanobody, a single-chain variable fragment (scFv), a tandem-di-scFv, a tandem-tri-scFv, a scFv-Fc, a (scFv-CH3)2 (also termed minibody), a (scFv- CH2-CH3)2 (also termed maxibody), a diabody, and a triabody.

[0118] In some embodiments, the antibody, or antigen-binding fragment thereof, is selected from the group comprising or consisting of an immunoglobulin single variable domain (ISVD) (such as, for example, a Variable Heavy domain of Heavy chain (VHH) or a nanobody) or single domain antibody (sdAb),. In a preferred embodiment, the antibody, or antigen-binding fragment thereof, is selected from the group comprising or consisting of an immunoglobulin single variable domain (ISVD) (such as, for example, a Variable Heavy domain of Heavy chain (VHH) or a nanobody) or a single domain antibody (sdAb).

[0119] The term “binding fragment”, as used herein, refers to a part or region of the antibody according to the present invention, which comprises fewer amino acid residues than the whole antibody. A “binding fragment” binds antigen and / or competes with the whole antibody from which it was derived for antigen binding (e.g., specific binding to antigen). Antibody binding fragments encompasses, without any limitation, single chain antibodies, Fv, Fab, Fab', Fab'-SH, F(ab)’2, Fd, defucosylated antibodies, diabodies, triabodies and tetrabodies.

[0120] “Single chain antibody”, as used herein, refers to any antibody or fragment thereof that is a protein having a primary structure comprising or consisting of one uninterrupted sequence of contiguous amino acid residues, including without limitation (1) single-chain Fv molecules (scFv); (2) single chain proteins containing only one light chain variable domain, or a fragment thereof that contains the three CDRs of the light chain variable domain, without an associated heavy chain moiety; and (3) single chain proteins containing only one heavy chain variable region, or a fragment thereof containing the three CDRs of the heavy chain variable region, without an associated light chain moiety.

[0121] An "immunoglobulin single variable domain" (ISVD) refers to a protein structure that retains the sequence FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 within a single domain. As a critical distinction, these ISVDs differ from conventional immunoglobulins in that the antigen-binding site is contained entirely within a single variable domain. This makes ISVDs unique in requiring only three CDRs for their antigen-binding activity, as opposed to the six involved in conventional antibody domains. ISVDs can include both light chain variable domains, such as VL sequences, or heavy chain variable domains like VH or VHH sequences. These structures are functionally capable of binding antigens as standalone units, simplifying their use in therapeutic and diagnostic applications. One significant application of ISVDs is in the development of heavy-chain-only antibodies, particularly those derived from camelid species. These antibodies, often referred to as Nanobodies, consist solely of VHH sequences, representing a subclass of ISVDs. These single-domain antibodies have garnered interest due to their exceptional stability, solubility, and ease of production. Nanobodies exemplify the structural and functional capabilities of ISVDs, which combine simplicity and efficacy in antigen targeting. The development of ISVDs, including Nanobodies, often involves humanization techniques to reduce immunogenicity when used in therapeutic contexts. Humanization entails modifying the framework regions of these antibodies to align moreclosely with human antibody sequences. This process may involve substituting amino acids within the framework regions without compromising antigen-binding functionality. Computational tools and databases are frequently employed to identify homologous human germline sequences, which guide the introduction of appropriate substitutions. For instance, humanized ISVDs can retain the antigen-binding activity of their non-human counterparts while significantly reducing the likelihood of adverse immune responses in patients. The design of such ISVDs often focuses on optimizing their framework regions to preserve their structural integrity and binding affinity. These efforts ensure the compatibility of ISVDs with the human immune system, making them viable candidates for therapeutic applications. Nanobodies, as a subclass of ISVDs, are characterized by their compact size and high binding affinities. These features make them particularly attractive for applications requiring precise targeting of antigens. The antigen-binding capabilities of Nanobodies are largely determined by their CDRs, with the CDR3 region playing a particularly crucial role. The diversity in CDR3 sequences among Nanobodies enables the recognition of a broad range of antigens, including those with complex epitopes. This versatility underpins the utility of Nanobodies in various therapeutic and diagnostic applications. The use of ISVDs extends beyond camelid-derived Nanobodies to include synthetic and engineered variants. These variants can be designed to exhibit specific properties, such as enhanced binding affinity or stability.

[0122] In some embodiments, the antibody or antigen-binding fragment thereof also encompasses a multivalent or multispecific binding agent, z.e., more than one, such as at least two, antibodies or binding fragments thereof, whether identical or different, being covalently linked together, directly or indirectly.

[0123] In the present invention, unless otherwise specified, the position of the complementary-determining regions (CDRs) is determined using the Kabat nomenclature.

[0124] Hence, in some embodiments, the molecular system further comprises at least one immunoglobulin single variable domain (ISVD) (such as, for example, a Variable Heavy domain of Heavy chain (VHH)) or single domain antibody (sdAb), that specifically binds to a cell surface of a bacteria, preferably bacteria of the genus Acinetobacter, more preferably bacteria of the species Acinetobacter baumannii. In some embodiments, the molecular system further comprises at least one ISVD (e.g., VHH) or sdAb that specifically binds to a proteinfrom bacteria, preferably bacteria of the genus Acinetobacter, more preferably bacteria of the species Acinetobacter baumannii. In some embodiments, the molecular system further comprises at least one ISVD (e.g., VHH) or sdAb that specifically binds to a surface protein from bacteria, preferably bacteria of the genus Acinetobacter, more preferably bacteria of the species Acinetobacter baumannii. In some embodiments, the molecular system further comprises at least one ISVD (e.g, VHH) or sdAb that specifically binds to the protein Omp25 from Acinetobacter baumannii.

[0125] In some embodiments, the antibody or the antigen-binding fragment thereof, preferably the ISVD (e.g, VHH) or sdAb, comprises the three following CDRs:- CDR1: GRVRSNT (SEQ ID NO: 8),- CDR2: AIKWIGTSTHYADSVKG (SEQ ID NO: 9), and- CDR3: AARYYSGFYLPAALAHEY (SEQ ID NO: 10).

[0126] In some embodiments, the antibody or the antigen-binding fragment thereof, preferably the ISVD (e.g, VHH) or sdAb, comprises CDRs having an amino acid sequence as set forth in SEQ ID NOs: 8-10 as described above with 1, 2, 3 or more amino acid(s) being substituted by a different amino acid. In some embodiments, the antibody or the antigen¬ binding fragment thereof, preferably the ISVD, sdAb, or VHH, comprises CDRs as described above having an amino acid sequence that shares at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%, or more identity with the corresponding amino acid sequence as set forth in SEQ ID NOs: 8-10.

[0127] In some embodiments, the antibody or the antigen-binding fragment thereof, preferably the ISVD (e.g, VHH) or sdAb, comprises or consists of an amino acid sequence having at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% of amino acid sequence identity with a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, preferably wherein the antibody or the antigen-binding fragment thereof, preferably the ISVD (e.g, VHH) or sdAb, comprises the three following CDRs:- CDR1: GRVRSNT (SEQ ID NO: 8),- CDR2: AIKWIGTSTHYADSVKG (SEQ ID NO: 9), and- CDR3: AARYYSGFYLPAALAHEY (SEQ ID NO: 10).

[0128] In some embodiments, the antibody or the antigen-binding fragment thereof, preferably the ISVD (e.g., VHH) or sdAb, comprises or consists of an amino acid sequence as set forth in the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3.

[0129] In some embodiments, the isolated antibody or antigen-binding fragment thereof, preferably the ISVD (e.g., VHH) or sdAb, is humanized.

[0130] In some embodiments, the humanized antibody or the antigen-binding fragment thereof, preferably the humanized ISVD (e.g., VHH) or sdAb, comprises CDRs having an amino acid sequence as set forth in SEQ ID NOs: 11-15 as described above with 1, 2, 3 or more amino acid(s) being substituted by a different amino acid. In some embodiments, the humanized antibody or the antigen-binding fragment thereof, preferably the humanized ISVD (e.g., VHH) or sdAb, comprises CDRs as described above having an amino acid sequence that shares at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%, or more identity with the corresponding amino acid sequence as set forth in SEQ ID NOs: 11-15.

[0131] In some embodiments, the humanized antibody or the antigen-binding fragment thereof, preferably the humanized ISVD (e.g., VHH) or sdAb, comprises or consists of an amino acid sequence having at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% of amino acid sequence identity with a sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, and SEQ ID NO: 15, preferably wherein the humanized antibody or the antigen-binding fragment thereof, preferably the humanized ISVD (e.g., VHH) or sdAb, comprises the three following CDRs: - CDR1: GRVRSNT (SEQ ID NO: 8),- CDR2: AIKWIGTSTHYADSVKG (SEQ ID NO: 9), and- CDR3: AARYYSGFYLPAALAHEY (SEQ ID NO: 10).

[0132] In some embodiments, the humanized antibody or the antigen-binding fragment thereof, preferably the humanized ISVD (e.g., VHH) or sdAb, comprises or consists of an amino acid sequence as set forth in the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, and SEQ ID NO: 15.

[0133] Hence, in some embodiments, the molecular system further comprises at least one ISVD (e.g., VHH) or sdAb comprising the three following CDRs:- CDR1: GRVRSNT (SEQ ID NO: 8),- CDR2: AIKWIGTSTHYADSVKG (SEQ ID NO: 9), and- CDR3: AARYYSGFYLPAALAHEY (SEQ ID NO: 10).

[0134] In some embodiments, the molecular system further comprises at least one ISVD (e.g., VHH) or sdAb having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, and SEQ ID NO: 15. In some embodiments, the molecular system further comprises at least one ISVD (e.g., VHH) or sdAb having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3. In some embodiments, the molecular system further comprises at least one ISVD (e.g., VHH) or sdAb having an amino acid sequence selected from the group consisting of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, and SEQ ID NO: 15.

[0135] According to some embodiments each of the antibodies is covalently bound to at least one of the pillararenes by means of a single bond, a coupling product, or a (C1-C4) alkyl, wherein the alkyl is optionally interrupted or terminated by at least one coupling product. Therefore, the bond between the antibody and the pillararene can either be reversible or non-reversible. In some embodiments, each of the antibodies is covalently bound to at least one of the pillararenes by means of a coupling product. In some preferred embodiments, the coupling product is N=CH (i.e., the bond is an imine bond). In some embodiments, each of the antibodies is covalently bound to one of the pillararenes (i.e., “to exactly one of the pillararenes”, in other words, no antibody can be bound to two or more pillararenes) by means of a single bond, a coupling product, or a (C1-C4) alkyl, wherein the alkyl is optionally interrupted or terminated by at least one coupling product. According to some preferred embodiments, each of the antibodies is covalently bound to at least one of the pillararenes by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product. In some embodiments, each of the antibodies is covalently bound to at least one of the pillararenes by means of a reversible coupling product. In some preferred embodiments, the reversible coupling product is N=CH (i.e., the bond is an iminebond). Thereby, each of the antibodies is covalently bound in a reversible way to at least one of the pillararenes.

[0136] According to some other embodiments each of the antibodies is covalently bound to at least one of the cores by means of a single bond, a coupling product, or a (C1-C4) alkyl, wherein the alkyl is optionally interrupted or terminated by at least one coupling product. Therefore, the bond between the antibody and the core (z.e., the bond between the antibodies and the DCF) can either be reversible or non-reversible.

[0137] In some embodiments, the binding moieties within said multivalent or multispecific agent may be directly linked, or fused by a linker or spacer. The composition or binding agent(s) as described herein may appear in a “multivalent” or “multispecific” form and thus be formed by bonding, chemically or by recombinant DNA techniques, together two or more identical or different binding agents. In some embodiments, the multivalent forms are formed by connecting the building blocks directly or via a linker, or through fusing the building block(s) with an Fc domain encoding sequence. Non-limiting examples of multivalent constructs include “bivalent” constructs, “trivalenf ’ constructs, “tetravalent” constructs, and so on. In some embodiments, the immunoglobulin single variable domains comprised within a multivalent construct are identical or different, preferably binding to the same or overlapping binding site. In another particular embodiment, the binding agent(s) of the invention are in a “multispecific” form and are formed by bonding together two or more building blocks or agents, of which at least one binds to one target protein, preferably to Omp25 from Acinetobacter baumannii, and at least one binds to a further target or alternative molecule, so when present in multispecific fusion, presenting a binding agent or composition that is capable of specifically binding both epitopes or targets, thus comprising binders with a different specificity. Non-limiting examples of multi-specific constructs include “bispecific” constructs, “trispecific” constructs, “tetraspecific” constructs, and so on. To illustrate this further, any multivalent or multispecific (as defined herein) form of the invention may be suitably directed against one or more different epitopes on the same antigen, or on epitopes of different surface proteins, or may be directed against two or more different antigens. Multivalent or multi¬ specific ISVDs of the invention may also have (or be engineered and / or selected for) increased avidity and / or improved selectivity for the desired bacterial interaction, and / or for any other desired property or combination of desired properties that may be obtained by the use of suchmultivalent or multispecific immunoglobulin single variable domains. Upon binding, said multi-specific or multivalent binding agent may have an additive or synergistic impact on the binding and / or therapeutic effect on its target, such as an increase in its potency or selectivity. In another embodiment, the immunoglobulin single variable domains, either in a monovalent, multivalent or multispecific form are VHHs or Nanobodies as included here as non-limiting examples. In some embodiments, the multivalent or multispecific binders or building blocks may be fused directly or fused by a suitable linker to each other, as to allow that the at least two binding sites can be reached or bound simultaneously by the multivalent or multispecific agent. Alternatively, at least one ISVD as described herein may be fused at its C -terminus to an Fc domain, for instance an Fc-tail of an Ig, resulting in an antigen-binding protein of bivalent format wherein two of said VHH-Ig Fes, or humanized forms thereof, form a heavy chain only- antibody-type molecule through disulfide bridges in the hinge region of the Fc part, called “Fc fusion” herein. In a specific embodiment, the multivalent or multispecific agent as described herein is an Fc fusion or an antibody. Another embodiment comprises a humanized ISVD specifically binding the target as described herein, comprised in a multivalent or multispecific agent, which may be provided as a humanized ISVD-IgG fusion, and which may further include but is not limited to the use of IgG humanization variants known in the art, such as C- terminal deletion of Lysine, alteration or truncation in the hinge region, LALA or LALAPG mutations as described, among other substitutions in the IgG sequence. In an alternative embodiment, the “Fc fusion” is designed by linking the C-terminus of such a bivalent or bispecific binder fused by a linker to an Fc domain, which then upon expression in a host forms a multivalent or multispecific-antibody-type molecule through disulfide bridges in the hinge region of the Fc part.

[0138] According to some embodiments, the antibody is prepared and characterized as described in the Examples herein, and as defined in unpublished patent application PCT / EP2024 / 067693 (VIB vzw et al.), in particular the Examples thereof.Alternative forms of the molecular system

[0139] All references to molecular systems according to the invention include references to salts, solvates, multi-component complexes and / or liquid crystals thereof. All references to molecular systems according to the invention include references to polymorphs and / or crystalhabits thereof. All references to molecular systems according to the invention include references to pharmaceutically acceptable prodrugs thereof All references to molecular systems according to the invention include references to isotopically-labelled compounds thereof, including deuterated compounds thereof

[0140] The molecular systems according to the invention may contain at least one asymmetric centre(s) and thus may exist as different stereoisomeric forms. Accordingly, all references to molecular systems according to the invention include references to all possible stereoisomers and includes not only the racemic compounds but the individual enantiomers and their non-racemic mixtures as well. When a compound is desired as a single enantiomer, such single enantiomer may be obtained by stereospecific synthesis, by resolution of the final product or any convenient intermediate, or by chiral chromatographic methods as each are known in the art. Resolution of the final product, an intermediate, or a starting material may be carried out by any suitable method known in the art. Bonds from an asymmetric carbon in compounds are generally depicted using a solid line ( - ),a solid wedgeor a dotted wedge ( > " HI). The use of either a solid or dotted wedge to depict bonds from an asymmetric carbon atom is meant to indicate that only the stereoisomer shown is meant to be included.

[0141] The molecular systems according to the invention may be in the form of pharmaceutically acceptable solvates. Pharmaceutically acceptable solvates of the compounds include hydrates thereof.

[0142] The molecular systems according to the invention may be in the form of pharmaceutically acceptable salts. Pharmaceutically acceptable salts of the compounds include the acid addition and base salts thereof. Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include the acetate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, bi sulphate / sulphate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulphate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate and xinafoate salts. Suitable base salts are formed from baseswhich form non-toxic salts. Examples include the aluminium, arginine, benzathine, calcium, choline, diethylamine, 2-(diethylamino)ethanol, diolamine, ethanolamine, glycine, 4-(2-hydroxyethyl)-morpholine, lysine, magnesium, meglumine, morpholine, olamine, potassium, sodium, tromethamine and zinc salts. Hemisalts of acids and bases may also be formed, for example, hemisulphate and hemicalcium salts. When the molecular systems according to the invention contain an acidic group as well as a basic group the compounds of the invention may also form internal salts, and such compounds are within the scope of the invention. When the molecular systems according to the invention contain a hydrogen-donating heteroatom (e.g., NH), the invention also covers salts and / or isomers formed by transfer of said hydrogen atom to a basic group or atom within the molecule. Pharmaceutically acceptable salts of molecular systems according to the invention may be prepared by one or more of these methods: (i) by reacting the molecular system with the desired acid; (ii) by reacting the molecular system with the desired base; (iii) by removing an acid- or base-labile protecting group from a suitable precursor of the molecular system or by ring-opening a suitable cyclic precursor, e.g., a lactone or lactam, using the desired acid; and / or (iv) by converting one salt of the molecular system to another by reaction with an appropriate acid or by means of a suitable ion exchange column. All these reactions are typically carried out in solution. The salt may precipitate from solution and be collected by filtration or may be recovered by evaporation of the solvent. The degree of ionization in the salt may vary from completely ionized to almost non-ionized.Pharmaceutical composition

[0143] This invention also relates to a pharmaceutical composition comprising a molecular system according to the invention, as described hereinabove, and at least one pharmaceutically acceptable carrier. According to a first embodiment, the pharmaceutical composition comprises the molecular system according to the invention as sole therapeutic agent. According to a second embodiment, the pharmaceutical composition further comprises at least another therapeutic agent. In some embodiments, the other therapeutic agent is an anti-infective agent. In some preferred embodiments, the other therapeutic agent is an antibiotic.Medical use

[0144] This invention also relates to a molecular system or a pharmaceutical composition according to the invention, as described hereinabove, for use as a medicament.

[0145] This invention also relates to a molecular system or a pharmaceutical composition according to the invention, as described hereinabove, for use in the treatment in the treatment of an infectious disease.

[0146] According to some embodiments, the infectious disease is a bacterial disease, a viral disease, a fungal disease, a mycobacterial disease or a parasitic disease. In some embodiments, the infectious disease is a bacterial disease. In some embodiments, the infectious disease is a viral disease. In one embodiment, the infectious disease is a fungal disease. In some embodiments, the infectious disease is a mycobacterial disease. In some embodiments, the infectious disease is a parasitic disease. In some embodiments, the infectious disease is a bacterial disease, a fungal disease or a mycobacterial disease. In some particular embodiments, the infectious disease is a bacterial disease or a mycobacterial disease. In some preferred embodiments, the infectious disease is a bacterial disease. In some embodiments, the bacterial disease is selected from pneumonia, bloodstream infections, meningitis, wound and surgical site infections and urinary tract infections.

[0147] According to some embodiments, the bacterial disease is caused by Gram-negative bacteria. In some embodiments, the bacterial disease is caused by bacteria selected from Acinetobacter baumannii, a member of the Enterobacterales (including, but not limited to, Escherichia coli, Shigella spp., Salmonella spp., Enterobacter spp., Klebsiella spp.), a member of the enterococci (including, but not limited to, Enterococcus faecalis and Enterococcus faecium), Pseudomonas aeruginosa, Neisseria spp., Staphylococcus aureus, a member of the streptococci (including, but not limited to, Streptococcus pneumoniae and Streptococcus pyogenes), Haemophilus influenzae, and mixtures thereof. In some preferred embodiments, the bacterial disease is caused by Acinetobacter baumannii bacteria.

[0148] Advantageously, the molecular system or a pharmaceutical composition according to the invention has an antibiofilm activity.

[0149] According to a first embodiment, the molecular system or a pharmaceutical composition according to the invention is administrated to the subject as sole therapeutic agent. According to a second embodiment, the molecular system or a pharmaceutical composition according to the invention is administrated to the subject in combination with at least another therapeutic agent. In some embodiments, the other therapeutic agent is an anti-infective agent. In some preferred embodiments, the other therapeutic agent is an antibiotic.

[0150] This invention also relates to the use of a molecular system or a pharmaceutical composition according to the invention, as described hereinabove, in the manufacture of a medicament for the treatment of an infectious disease. This invention also relates to a method for the treatment of an infectious disease in a subject in need thereof, comprising a step of administrating to said subject a therapeutically effective amount of a molecular system or a pharmaceutical composition according to the invention, as described hereinabove.Medical devices

[0151] This invention also relates to a medical device comprising a molecular system or a pharmaceutical composition according to the invention, as described hereinabove. According to some embodiments, the medical device is a prosthesis, an implant, a stent, or a surgical tool.Non-medical use

[0152] This invention also relates to the non-therapeutic use as antibiofilm agent of a molecular system or a pharmaceutical composition according to the invention, as described hereinabove.

[0153] This invention also relates to a non-therapeutic antibiofilm method comprising a step of applying on a surface or item a molecular system or a pharmaceutical composition according to the invention, as described hereinabove. According to some embodiments, the surface or item is not part of a human or animal body. For example, the surface may be part of a medical device (e.g., the surface of an operating table) or the item may be a medical device (e.g., a surgical tool), but no patient whatsoever is treated by the molecular system or a pharmaceutical composition.

[0154] According to some embodiments, the biofilm comprises Gram-negative bacteria as described hereinabove. In some preferred embodiments, the biofilm comprises Acinetobacter baumannii bacteria.

[0155] This invention also relates to the use of a molecular system as described hereinabove for water treatment. According to one embodiment, the molecular system is added in and / or contacted with the water to be treated.

[0156] This invention also relates to the use of a molecular system as described hereinabove for the treatment and / or prevention of contamination of a recipient or a packaging by at least one microorganism, for example, in the food industry.

[0157] This invention also relates to the use of a molecular system as described hereinabove for the treatment and / or prevention of contamination of building (e.g., a greenhouse) by at least one microorganism, for example, in the agriculture field.Manufacturing process

[0158] The molecular system according to the invention, as described hereinabove, can be prepared by any suitable synthetic method known in the art.

[0159] This invention also relates to a process for manufacturing a molecular system according to the invention, as described hereinabove, wherein the process comprises the following steps:(a) a step of reacting a plurality of cores as defined herein with a plurality of connectors as defined herein, thereby obtaining a base dynamic constitutional framework (“base DCF”); then(b) a step of reacting the base DCF with a plurality of precursors of a functional head as defined herein, thereby obtaining a functionalised dynamic constitutional framework (DCF);(c) providing a host-guest complex comprising at least one pillararene as defined herein and at least one anti-infective agent as defined herein, wherein each of the anti-infective agent(s) is hosted in one of the pillararene(s);(d) a step of reacting the functionalised dynamic constitutional framework (DCF) with said host-guest complex, thereby obtaining the molecular system; and(e) optionally, contacting and / or reacting the molecular system with a plurality of immunoglobulin single variable domains (ISVDs), thereby obtaining thereby obtaining a molecular system comprising at least one ISVD.

[0160] Although the above process presented the steps in a particular order, a process wherein the steps follow an alternative order is also possible, and is also within the scope of the present invention. Moreover, the numbering of the steps is not to be constructed as a limitation of the claimed scope. However, step (a) is as a rule carried out before step (b), so that the base DCF can be obtained with the desired dynamic structure, before being functionalised by the functional heads.

[0161] According to some embodiments, the host-guest complex of step (c) is obtained by a step (c-1) of contacting a pillararene as defined herein with an anti-infective agent as defined herein, thereby obtaining the host-guest complex. In these embodiments, the host-guest complex is prepared before being included in the molecular system.

[0162] In some embodiments, the host-guest complex of step (d) is replaced by a pillararene as defined herein, and step (c-1) as defined herein is instead carried out after step (d) or step (e), thereby obtaining the molecular system. In these embodiments, the host-guest complex is prepared in situ, after the pillararene is included in the molecular system

[0163] According to some embodiments, the molecular system is obtained by reaction of the functionalised-DCF as described herein with the host-guest complex as described herein.

[0164] According to some embodiments, at step (d) the molecular system is obtained by reaction of a functionalised-DCF as defined herein with at least one host-guest complex as defined herein; preferably the reaction between at least one coupling function of the functionalised-DCF (e.g., aldehyde) and at least one coupling function of the host-guest complex e.g., NH2); more preferably the reaction between exactly one coupling function of the functionalised-DCF and exactly one coupling function of the host-guest complex (thereby, each core of the functionalised-DCF is bound only once to a given host-guest complex).

[0165] In some embodiments, at step (d) the initial moral ratio between the functionalised-DCF and the host-guest complex(es) ranges from about 0.0033 to 0.82, preferably the initial moral ratio is about 0.05. In some embodiments, the initial equivalent moral ratio between the functionalised-DCF and the host-guest complex(es) ranges from about 0.033 to 8.2, preferably the initial equivalent moral ratio is about 0.5. By “equivalent moral ratio” it is meant the molar ratio taking into account the number of free functions (i.e., the functions available for reaction) in the reactants. For example, when the functionalised-DCF has one free aldehyde group and the host-guest complex has ten free amine groups, an initial molar ratio of about 0.05 corresponds to an initial equivalent molar ratio of about 0.5.BRIEF DESCRIPTION OF THE DRAWINGS

[0166] Figure 1. NbH7 binds the cell surface of AB5075-VUB-itrA::ISAba13 (AB5075C-). Fluorescence micrographs of NbH7 and a control Nb (CtrlNb) tested on AB5075-VUBC- in exponential and stationary phase are shown. Per condition, from left to right, the phase contrast, GFP, DL650 and an overlay image are shown.

[0167] Figure 2. Omp25 is the target of NbH7. Fluorescence micrographs of the binding ability of NbH7 on the AB5075C- strain and the derivative where the omp25 gene was deleted, namely AB5075C-Aomp25.

[0168] Figures 3A-3B. Biofilm growth inhibition assay. Figures 3A-3B are a combination of graph that shows the absolute values of Colony-forming units (CFUs), in response to individual components (Fig. 3 A) and combinations (Fig. 3B) treatment.

[0169] Figures 4A-4B. Biofilm growth inhibition assay. Figures 4A-4B are a combination of graph that shows the Relative logarithmic values of CFU compared to the control test in response to individual components (Fig. 4A) and combinations (Fig. 4B) treatment.

[0170] Figure 5. Optimization of combination composition for DCF-C-APAl-Lev by antimicrobial checkboard assay. The graph indicates synergy between the components. The optimal combination (DCF0: C: APAl: Lev = 1:1:0.05:0.1 by molar) is highlighted with a red circle.

[0171] Figure 6. MIC of AHPA2-Lev and DCF-C-AHPA2-Lev combinations in compared with DCF-C and AHPA2 alone. This graph illustrates the synergistic effect of combinations DCF-C-AHPA2-Lev and AHPA2-Lev by decreasing of MIC values.EXAMPLES

[0172] The present invention is further illustrated by the following examples.Example 1: Preparation of the molecular systemMaterials

[0173] All reagents and chemicals used in this study were purchased from Sigma-Aldrich, Across, or TCI Europe at ACS grade and used without purification. Deuterated solvents (CDCl3, D2O, CD3OD, and (CD3)2SO for NMR analysis were purchased from Cambridge Isotopes Laboratories (Eurisotop). All reactions were carried out in standard glassware. Benzene-l,3,5-tricarboxaldehyde 1, poly-(ethylene glycol)-bis(3-aminopropyl) terminated (Mn ~1500 Da) 2 were purchased from TCI Europe.DCF synthesis

[0174] To a solution of 1,3,5-Benzenetryaldehyde (BTA) in acetonitrile, was added polyethylene glycol) bi s(3 -aminopropyl) (1: 1 molar ratios), and the mixture was stirred for 2- 3 days at room temperature under an inert atmosphere (N2). Then, the solvent was removed under vacuum, and the product was immediately dissolved in MilliQ water at a concentration of 2 mg of TBA as a Stock Solution (DCFO). To a volume of DCFO containing 5 mg BTA (0,03 Immol), a third building block (functional module) was added (DCFO: Components: 1 or 1:2 by molar). After adding MilliQ water to obtain a final volume of 5 mL, the mixture was stirred at room temperature for at least 48-72 hours to obtain the final solution of DCFs.

[0175] List of functional heads (FH 3): branched polyethylene imine 800 Da - 3 A, branched polyethylene imine Mw ~ 2000 Da solution 50% in water- 3B, aminoguanidine hydrochloride - 3C, Isonicotinic acid hydrazide - 3G, Nicotinic acid hydrazide - 3H, nicotinic Pyrazine-2-carbohydrazide-3M, Girard’s Reagent P - 3N, Girard's reagent T - 30, 2 -Furoic hydrazide - 3U, 4-(hydrazinocarbonil) benzoic acid - 3W.Pillar[5]arene synthesisSynthesis of APA 1 (DMS T 14)

[0176] Synthesis of DMST6: 16g of K2CO3 (115 mmol) was added to the solution of 1.6-dibromohexane (8.2 mL, 50.6 mmol) in acetone (80 mL). The mixture was refluxed (60-65 °C) for 30 min under the inert atmosphere (argon). 40 mL of Hydroquinone solution in acetone(2.53 g, 23 mmol) was added dropwise, and the resulting reaction mixture was refluxed for 48 hours, reaction mixture was cooled down to room temperature, filtrated, and evaporated. The crude product was dissolved in DCM (50 ml), washed with water and brine, and dried over MgSO4. Purification was done by column chromatography on Silica with Cy: DCM=2: 1 eluent system. The final product was obtained as a white powder (3 g, 30%)

[0177] Synthesis of DMST8: The mixture of DMST6 (3.15 g, 7.22 mmol) and paraformaldehyde (0.685 g, 22.81 mmol) in DCE (50 mL) was stirred for 10 min at room temperature. 1 ml of BF3 EtzO (1.15 g, 14.44 mmol) was added dropwise under argon atmosphere. The reaction mixture was stirred for 2 h at room temperature. Water (50 mL) was added to quench the reaction. After that, the mixture was filtrated, the organic layer was separated, washed with K2CO3, water, and brine, and evaporated with rotavap. The crude product was purified with column chromatography (silica, Cy: DCM=l:l), and white powder was obtained (1.49 g, 46%)

[0178] Synthesis ofDMST13: 2 g of DMST8 (0.9 mmol) was dissolved in dry DMF (50 mL). Sodium azide (1.16 g, 18 mmol) was added under argon, and the reaction mixture was stirred for 24 hours at room temperature. After that, the solvent was evaporated under a vacuum and dissolved in DCM (25 mL). The solution was washed with water (25mL x3) and brine and dried with MgSO4. Evaporation of solvent using rotavap gave 1.68 g of resulting product as white solid (quant. Yield).

[0179] Synthesis ofDMST14: 500 mg of DMST13 (0.27 mmol) was dissolved in dry MeOH (20 mL). Palladium on Carbon (50 mg, 10%) was added under the argon atmosphere. After that, Hydrogen gas was added to the system, and the reaction mixture was stirred for 2 days. The reaction mixture was filtered and evaporated, giving 428 mg of white solid (quant. Yield). ‘HNMR (CDCh): 5 6.84 (H-8, s, ArH, 10H), 3.91-3.81 (H-6, br, OCH2, 20H), 3.70 (H-7, s, ArCHz Ar, 10H), 2.44 (H-l, br, CH2-NH2, 20H), 1.81-1.60 (H-2, br, CHz, 20H), 1.44-1.28 (H- 3, H-4, H-5, br, CHz, 60H) ppm.13C NMR (CDCl3): δ 149.65 (C-8), 128.28 (C-9), 114.38 (C-10), 68.20 (C-6), 41.29 (C-1), 32.53 (C-2), 29.61 (C-5), 29.20 (C-7), 26.53 (C-3), 26.31 (C-4) ppm. HRMS: Calculated 1601.2318; Found 1603.2395.Synthesis of APA2 (DMST22)

[0180] Synthesis of DMST10: 16g of K2CO3 (115 mmol) was added to the solution of Propargyl bromide (3.82 mL, 6 g, 50.6 mmol) in acetone (80 mL). The mixture was refluxed (60-65 °C) for 30 min under the inert atmosphere (argon). 40 mL of Hydroquinone solution in acetone (2.53 g, 23 mmol) was added dropwise, and the resulting reaction mixture was refluxed for 48 hours, reaction mixture was cooled down to room temperature, filtrated, and evaporated. The crude product was dissolved in DCM (50 ml), washed with water and brine, and dried over MgSO4. Purification was done by column chromatography on Silica with Cy: DCM=2:l eluent system. The final product was obtained as a white powder (3 g, 66%)

[0181] Synthesis of DMST12: The mixture of DMST10 (1.25 g, 6.713 mmol) and paraformaldehyde (0.6 g, 22.81 mmol) in DCE (25 mL) was stirred for 10 min at room temperature. 1.91 ml of BF3 EtzO (1.66 g, 13.42 mmol) was added dropwise under argon atmosphere. The reaction mixture was stirred for 2 h at room temperature. Methanol (25 mL) was added to quench the reaction. After that, the mixture was filtrated and evaporatedwith rotavap. The crude product was purified with column chromatography (silica, Cy: DCM=l:l), and white powder was obtained (505 mg, 38%).

[0182] Synthesis of DMST21: 50 mg of DMST12 (0.05 mmol, 1 eq.) was dissolved together with 274.3 mg of N3-PEG2-NH-Boc (1 mmol, 20 eq.) in 1.5 mL of DCM and stirred at room temperature under argon atmosphere. 16 mg of CuSCh (0.2 eq x 10) in water (0.75 mL) and 66 mg of Sodium Ascorbate (0.66 eq x 10) in water (0.75 mL) were added to the reaction and stirred overnight. The reaction mixture was diluted in 25 mL of water and 25 mL of DCM, and the organic layer was separated. After that, the solution was dried over MgSO4, filtrated, and evaporated using rotavap. Purification was performed by column chromatography (silica, DCM: MeOH=9:l) with the resulting product as a pale yellow oil (34 mg,60%). NMR (500 MHz, CHLOROFORM-D) δ 7.97(t, H-4, 10H), 6.94 (m, H-1, 10H), 5.25-4.83 (m, H-5, 20H), 4.52(s, H-3, 20H), 3.85 (s, H-2, 10H), 3.71-3.43-3.24 (m, H-6-H-10, 100H), 1.40 (s, H-11, 90H).13C NMR (126 MHz, CHLOROFORM-D) δ 156.09 (C-14), 149.73 (C-1), 144.09 (C-6), 128.77 (C-7), 124.25 (C-3), 116.15 (C-2), 79.28 (C-15), 70.53 (C-5), 70.29, 70.17, 69.49 (C-9 - C-12), 62.08 (C-4), 50.19 (C-8), 40.35 (C-13), 28.52 (C-16).

[0183] Synthesis of APA2 (DMST22): To a solution of DMST21 (34 mg, 0.01 mmol) in DCM (1.0 mL) at room temperature, TFA (0.5 mL) was added. The reaction mixture was stirred at RT for 2 h, after which the mixture was co-distilled with DCM 4-5 times in vacuo, which gave the resulting product a yellow oil (35 mg, quant, yield).1H NMR (500 MHz, CHLOROFORM-D) δ 8.06 (s, H-4, 10H), 6.84 (s, H-1, 10H), 4.76 - 4.63(m, H-5, 20H), 4.49 (s, H-3, 20H), 3.86 (s, H-2, 10H), 3.72-3.42 (m, H-6 - H-9, 80H) 3.08(m, H-10, 20H).13C NMR (126 MHz, METHANOL-D4 - CHLOROFORM-D) δ 149.39 (C-1), 143.84 (C-6), 128.70 (C-7), 125.02 (C-3), 114.91 (C-2), 69.81(C-9), 69.61(C-5) 68.90(C-12), 66.59(C-10, C-11), 61.29 (C-4), 50.15(C-8), 39.36(C-13). HRMS: C125H190N40O30 Exact Mass 2731.4572 (Calculated); 2733.4750 ([M+H]+, Found).Synthesis of AHPA2[Chemical scheme caption text for Synthesis of AHPA2 diagram]RC30 RC18

[0184] Synthesis of DMST10: 16g of K2CO3 (115 mmol) was added to the solution of Propargyl bromide (3.82 mL, 6 g, 50.6 mmol) in acetone (80 mL). The mixture was refluxed (60-65 °C) for 30 min under an inert atmosphere (argon). 40 mL of Hydroquinone solution in acetone (2.53 g, 23 mmol) was added dropwise, and the resulting reaction mixture was refluxed for 48 hours. The reaction mixture was cooled down to room temperature, filtered, and evaporated. The crude product was dissolved in DCM (50 ml), washed with water and brine, and dried over MgSO4. Purification was performed by column chromatography on silica using a Cy: DCM = 2:1 eluent system. The final product was obtained as a white powder (3 g, 66%)

[0185] Synthesis of DMST12: The mixture of DMST10 (1.25 g, 6.713 mmol) and paraformaldehyde (0.6 g, 22.81 mmol) in DCE (25 mL) was stirred for 10 min at roomtemperature. 1.91 ml of BF3 EtzO (1.66 g, 13.42 mmol) was added dropwise under an argon atmosphere. The reaction mixture was stirred for 2 h at room temperature. Methanol (25 mL) was added to quench the reaction. After that, the mixture was filtered and evaporated with rotavap. The crude product was purified by column chromatography (silica, Cy: DCM=l:l), yielding a white powder (505 mg, 38%).

[0186] Synthesis ofRC14 To a solution of ethyl 5-bromopentanoate (1g, 4.8 mmol,l equiv.) in dry DMF (4 mL) was added NaN3(780 mg, 12 mmol, 2.5 equiv.) and stirred at room temperature overnight. Then, the solution was diluted with EtzO (80 mL), washed with H2O (2 x 60 mL), brine (60 mL), dried over MgSO4, and evaporated to dryness to afford a yellow oil (818 mg, 4.78 mmol, 99%).!H NMR (500 MHz, CHLOROFORM- D) 54.24-4.12 (m, 2H), 3.39-3.29 (m, 2H), 2.43-2.31 (m, 2H), 1.81-1.61 (m, 4H), 1.37- 1.24 (m, 3H).13C NMR (126 MHz, CHLOROFORM-D) 5 173.3, 60.5, 51.2, 33.8, 28.4, 22.3, 14.4.

[0187] Synthesis of RC18: 100 mg of DMST12 (0.05 mmol, 1 eq.) was dissolved together with 343 mg of RC14 (1 mmol, 20 eq.) in 10 mL of DCM and stirred at room temperature under an argon atmosphere. 125 mg of CuSO4·5H2O (0.5 eq x 10) in water (5 mL) and 198 mg of Sodium Ascorbate (1 eq x 10) in water (5 mL) were added to the reaction, which was stirred for 3 days. The reaction mixture was diluted in 25 mL of water and 25 mL of DCM, and the organic layer was separated. After that, the solution was dried over MgSO4, filtered, and evaporated using rotavap. Purification was performed by column chromatography (silica, DCM : Acetone = 1:1) with the resulting product as a pale yellow oil (156 mg, 58%).1H NMR (500 MHz, DMSO-D6) δ 8.19 (s, 10H), 6.89 (s, 10H), 4.95 (s, 10H), 4.72 (s, 10H), 4.25-3.96 (m, 20H), 3.61 (s, 10H), 2.23-2.21 (m, 20H), 2.20 (m, 20H), 1.72 (m, 20H), 1.42-1.39 (m, 20H), 1.12-1.09 (m, 30H).13C NMR (126 MHz, DMSO-D6) δ 172.94, 149.50, 144.09 128.73, 124.46, 115.26, 62.18, 60.21, 49.55, 40.35, 33.29, 29.53, 21.91, 14.54.

[0188] Synthesis of RC30: RC18 (250 mg, 0.092 mmol, 1 eq.) and hydrazide monohydrate (461 mg, 9.2 mmol, 100 eq.) were dissolved in dry EtOH (15 mL). The solution was refluxed for 48 h under an argon atmosphere and then concentrated under reduced pressure. The crude was dissolved in methanol, and excess TFA was added, stirred for 1 h, and centrifuged. The solution was evaporated, dissolved in water, andlyophilized to remove excess TFA and obtain the pink oil ( 125 mg, 50%)1H NMR (500 MHz, DMSO-D6) 5 8.13 (s, 10H), 6.92 (s, 10H), 5.02 (s, 10H), 4.84 (s, 10H), 4.31 (m, 20H), 3.64 (m, 10H), 2.20 (m, 20H), 1.84 (m, 20H), 1.52 (m, 20H).13C NMR (126 MHz, DMSO-D6) 6 159.87, 150.18, 144.25, 129.27, 124.54, 116.60, 63.20, 49.77, 40.35, 33.06, 29.70, 22.45.Host-guest complexesHOST GUESTSPreparation

[0189] A series of solutions with different ratios Host: Guest in DMSO-D6were prepared: 2:1, 1:1, 1:2 by molar ratios. The concentration of APA1 (DMST14) was fixed and was 4 mM.Analytical methods

[0190] 1H-NMR spectroscopy was used to investigate Host-Guest interaction between Pillararene APA1 (Host) and Antibiotics Lev, Lin and Cef (Guests).1H-NMR spectra were recorded on a JEOL ECX 400 or 500.Results

[0191] The binding affinity of APA1 with antibiotic Lev was tested by1H NMR titration experiments carried out by adding increasing amounts of Lev (0.5, 1.0 and 2.0 equiv.) to a 1.0 mM DMSO-D6solution of APA1. In agreement with the formation of an inclusion complex undergoing fast association / dissociation on the NMR time-scale, the hydrogen resonances of the pyridinone ring from the condensed heterocycle of the guest underwent substantial upfield shifts as a consequence of the magnetic shielding of the host molecule (Δδ = − 0.1 ppm for the H-8 hydrogen atom, for APA1:Lev = 1:2).Preparation of molecular systems [DCF-Pillar[5]arene combinations]

[0192] To prepare the molecular systems, the stock solution of DCF0 or DCF with functional head in MilliQ water was mixed with a solution of Pillararene alone or Pillararene / Antibiotic complex in DMSO in molar ratios BTA: Pillararene=l:l or 1:2 (resulting content of DMSO was 4%) and stirred for 48-72 hours at room temperature. The resulting solution was sonicated (if necessary) for homogenization.Material characterisation

[0193] DLS Analysis: 80 pL solution of sample (DCF0, DCF with Functional Heads, Pillararene, Pillararene / Antibiotic, or their combinations) was mixed with 420 pL MilliQ water. This solution was introduced into a disposal microcuvette (d=10mm) and analysed using DelsaTM Nano C Particle Analyzer with the following settings: Accumulation time 70s, scattering angle 165°, correlation method D, attenuator 1-100 %, pinhole 100 pm. The reported hydrodynamic diameter values are based on the intensity distribution data.

[0194] TEM: Transmission electron microscopy (TEM) images were obtained using a Philips Tecnai 10 microscope operating at 80 kV. Samples were deposited onto a copper grid as an aqueous solution and dried at room temperature.

[0195] AFM: The samples were imaged using a Bruker Nanoscope operated in tapping mode under ambient conditions. Silicon cantilever tips with a gold reflecting coating, a resonance frequency of 140-390 kHz, a force constant of 3.1-37.6 N m-1, and a tip curvature radius of 10 nm were used. Sample preparation: A 10-pL aliquot solution wasdeposited on freshly cleaved mica substrates and dried in air at room temperature prior to imaging.Results

[0196] The binding affinity of APA1 with antibiotic Lev was tested by NMR titration experiments carried out by adding increasing amounts of Lev (0.5, 1.0 and 2.0 equiv.) to a 1.0 mM DMSO-D6solution of APA1. In agreement with the formation of an inclusion complex undergoing fast association / dissociation on the NMR time-scale, the hydrogen resonances of the pyridinone ring from the condensed heterocycle of the guest underwent substantial upfield shifts as a consequence of the magnetic shielding of the host molecule (Δδ = − 0.1 ppm for the H-8 hydrogen atom, for APA1:Lev = 1:2).Example 2: Biological evaluation - Antimicrobial activity 64. baumanniiMaterials and MethodsAntimicrobial components

[0197] The following substances were investigated for antimicrobial properties:- Dynamic constitutional framework consisting of Benzene-l,3,5-tricarbaldehyde and Poly-(ethyleneglycol)-bis(3-aminopropyl) terminated-PEG (1:1 molar ratio) - DCF0 core;O <noQDCF0- Pillar[5]arenes: Decaaminosubstituted Pillar[5]arene APA1 (DMST14) and Decaamino-PEG-substituted Pillar[5]arene APA2 (DMST22) as represented hereinabove.- Functional heads: Aminoguanidine hydrochloride - 3C;HCI NH2HN ' NHI NH '2C- Antibiotic: Levofloxacin - Lev.o oMulticomponent systems

[0198] Several 2-component combinations were evaluated, namely DCF-C, DCF-APA1, DCF-APA2, APAl-Lev, and APA2-Lev.

[0199] In addition to 2-component combinations, four complex multi-component systems were evaluated.A) 4-component combinations - the combined system of dynamic polymer with drug delivery complex (DCF-C-APAl-Lev, DCF-C-APA2-Lev).B) 5-component combinations - the combined system of dynamic polymer with drug delivery complex and Nanobody (DCF-C-APAl-Lev-NB (or NBO), DCF-C-APA1- Lev-FNB).Screening of multicomponent systems for antimicrobial activity against A. baumannii

[0200] Sample solutions of individual components and multi-component systems were placed in a 96-well plate (50 pL each) and diluted with MilliQ water (1:2 dilution with 4% DMSO solution) to obtain seven concentrations, followed by adding a bacterialculture of A. baumannii (150 pL). Bacterial growth monitoring was conducted at 37°C with periodic readings of optical density OD600over 24 hours.Minimum inhibitory concentration and Fractional inhibitory concentration index calculations

[0201] Minimum inhibitory concentration (MIC) was determined as the minimum concentration of antimicrobial agent or its combination, inhibiting bacterial growth during 24-hour monitoring time.

[0202] The fractional inhibitory concentration index (FICI) was calculated as the ratio of the MIC of the component in the complex system to the MIC of an individual substance. To determine the overall effect of the combination, the sum of FIC for each system was compared with literature data (Table 1).Table 1. Matching of the Sum of FICI and Combination effect (MELETIADIS, J. et al., Antimicrobial agents and chemotherapy, Vol. 54, No. 2, 2010).Σ FICI Effect< 0.5 SYNERGY0.5-1.0 ADDITIVE1.0-4.0 INDIFFERENCE> 4.0 ANTAGONISMResults

[0203] The present disclosure shows the investigation of the antimicrobial activity of various pillar[5]arene-based multi-component DCF systems against the multidrug¬ resistant pathogen Acinetobacter baumannii. To evaluate the potential of these systems, minimum inhibitory concentration (MIC) and fractional inhibitory concentration index (FICI) values were measured for individual components and for combinations of components. The effect of each combination was classified as synergistic, additive, indifferent, or antagonistic based on FICI results (Table 1).Activity of individual components

[0204] Table 2 summarizes the MIC values of individual antimicrobial components from previous investigations. While several components, such as APA1 and levofloxacin (Lev), demonstrated moderate activity (MIC of 124 pg / mL and 56 pg / mL, respectively), others like DCFO and APA2 required higher concentrations to inhibit bacterial growth. Importantly, nanobodies (both functionalized and non-functionalized) and aminoguanidine hydrochloride (C) showed no significant antimicrobial activity on their own (MIC > 900 pg / mL for nanobodies and > 684 pg / mL or >10944 pg / mL for C), suggesting that their primary role might be as enhancers in multi-component systems rather than as direct antimicrobial agents.Table 2. MIC values of individual building blocksMICComponent Comment(pg / mL)DCFO 500 Activity at high concentrationC > 684 No antimicrobial activity..APA1 124 Activity at moderate concentrationLev 56 Activity at moderate concentrationActivity of the multi-component systems

[0205] Table 3, Table 3 bis and Table 4 provides an overview of MIC and FICI values for various multi-component systems. The analysis revealed several key findings.

[0206] Strong Synergistic Systems: The two-component system DCF-C exhibited strong synergy (EFICI = 0.38), with a notable decrease in MIC values for both DCFO (125 pg / mL) and C (86 pg / mL). This reduction suggests that the DCF matrix enhances the antimicrobial action of aminoguanidine hydrochloride, possibly by facilitating interaction with the bacterial cell membrane.

[0207] Pillar[5]arene-Antibiotic Complexes: The APAl-Lev and APA2-Lev systems exhibited synergy or partial synergy, with APAl-Lev achieving an FICI of 0.5 (synergistic) and APA2-Lev showing partial synergy (EFICI = 0.60). Notably, the MICsof both APA1 and Lev decreased significantly in the presence of each other, indicating a potential mutual enhancement of antimicrobial properties (Figure 2).

[0208] Higher-order Combinations: The four-component system DCF-C-APAl-Lev demonstrated strong synergy (EFICI = 0.44), with considerable MIC reductions across components, particularly for APA1 and Lev.Table 3. Minimal inhibition concentration MIC (pg / mL and mM) and Fractional inhibition concentration index FICI of combinations.MIC MIC MIC MICCombination (Comp 1) (Comp 1) (Comp 2) (Comp 2)(pg / mL) (mM) (pg / mL) (mM) DCF B 250 1.54 3085 1.54 DCF C 125 0.77 86 0.78 DCF APA1 500 3.08 248 0.15 DCF APA2 500 3.08 1199 0.44 APA1 Lev 31 0.02 14 0.04APA2 Lev 248 0.09 56 0.15 Table 3 (continued).FICI FICICombination ΣFICI(Comp 1) (Comp 2)DCF B 0.49 1.00 1.49DCF C 0.25 0.13 0.38DCF APA1 0.99 2.00 2.99DCF APA2 0.99 1.00 1.99APA1 Lev 0.25 0.25 0.50APA2 Lev 0.21 1.00 1.21Table 3bis Minimal inhibition concentration MIC (pg / mL and mM) and Fractional inhibition concentration index FICI of combinations.MIC MIC MIC MICCombination (Comp 1) (Comp 1) (Comp 2) (Comp 2)(pg / mL) (mM) (pg / mL) (mM) DCF B 250 1.54 3085 1.54 DCF C 125 0.77 86 0.78 DCF APA1 500 3.08 248 0.15DCF APA2 500 3.08 1199 0.44APA1 Lev 31 0.02 14 0.04APA2 Lev 124 0.045 28 0.075 Table 3bis (continued)FICI FICICombination ΣFICI(Comp 1) (Comp 2)DCF B 0.49 1.00 1.49DCF C 0.25 0.01 0.26DCF APA1 0.99 2.00 2.99DCF APA2 0.99 1.00 1.99APA1 Lev 0.25 0.25 0.50APA2 Lev 0.10 0.50 0.60T ble 4. Summary table of the results of the analysis of antimicrobial activity (MICs and FICIs) of multicomponent systems (A. baumannii)MICSystem Component FICI Σ FICI Effect (ig / mLLDCF0 125 0.25 Strong DCF-C 0.38C 86 0.13 synergy APA1 31 0.25APAl-Lev 0.50 Synergy Lev 14 0.25APA2 124 0.10 Partial APA2-Lev 0.60Lev 28 0.50 synergy DCF0 63 0.13DCF-C- C 43 0.060.44 Synergy APAl-Lev APA1 16 0.13Lev 7 0.13DCF0 125 0.25DCF-C- C 86 0.13 Partial 0.68APA2-Lev APA2 62 0.05 synergy*Lev 14 0.25* Comment: the main effect comes from DCF-C.

[0209] These results show the antimicrobial activity and synergistic effect of multicomponent DCF systems against Acinetobacter baumannii based on MIC determination and FICI calculation, and confirm that pillar[5]arene-based multi¬ component DCF systems exhibit significant antimicrobial activity against A. baumannii, particularly when combined with APA1 and Lev. The identified strong synergy in DCF-Cand DCF-C-APAl-Lev systems underscores the potential of DCF as a flexible and potent platform for enhancing the activity of traditional antimicrobials.

[0210] It is noteworthy that the total concentrations of compounds is reduced when using the 4-components system instead of the 2-component systems. Therefore, two times less amount of DCF-C-APAl-Lev is required in order to obtain a comparable antibacterial effect as DCF-C and APAl-Lev separately.Example 3: Biological evaluation - Antibiofilm activityMaterials and MethodsCompounds and combinations for antibiofilm assays

[0211] Six individual compounds and six combinations were chosen for antibiofilm assays: Dynamic polymer DCFO; Charged heads Aminoguanidine 3C; Pillararenes APA1 and APA2; Antibiotic Levofloxacin Lev (see Example 2).

[0212] For subsequent studies of the antibiofilm activities of combinations, it was proposed to use solution concentrations equal to 4x the combinations' MIC (Table 5, MICs of the DCF-C and APAl-Lev combinations are reference).Table 5. The concentration of tested compounds and combinationsCompound / C(l) C (2) Comp 1 Comp 2Combination (pg / mL) (pg / mL) DCF-C DCFO 500 C 344APAl-Lev APA1 124 Lev 56 APA2-Lev APA2 248 Lev 56 DCF-C-APAl-Lev DCFO 500 C 344DCF-C-APA2-Lev DCFO 500 C 344Table 5 (continued).Compound / C (3) C (4) Comp 3 Comp 4Combination (pg / mL) (pg / mL)DCF-C - - - -APAl-Lev - - - - APA2-Lev - - - - DCF-C-APAl-Lev APA1 124 Lev 56DCF-C-APA2-Lev APA2 248 Lev 56Antibiofilm assay setup

[0213] To allow the initial adhesion, a diluted suspension of bacteria (Acinetobacter baumannii strain AB5075) was added to a 96-well plate for 4 h. The medium was removed, and a mixture of fresh medium (low salt LB) and compound was added. The 96-well plate was incubated for another 20 h. The medium with compounds was removed, and bacteria were detached to perform a CFU count.Results

[0214] Table 6 and Table 7 present the results of the antibiofilm activity of multicomponent systems. Some 2-component systems, such as DCF-C, APAl-Lev, and APA2-Lev, have already been tested and show good reproducibility. Also, complex dynamic systems consisting of 4 components, such as DCF-C-APAl-Lev and DCF-C-APA2-Lev, show enhanced antibiofilm properties compared to individual two- component systems tested in this experiment: 4-component combination DCF-C-APA1-Lev has -3.49 of AloglO(CFU) compared to control test, while DCF-C and APAl-Lev have -1.58 and -1.87 respectively; 4-component combination DCF-C-APA2-Lev has a similar effect, the difference in loglO of CFU counts compared to untreated sample is - 5.2 with result of 100% reduction of absolute value of colony forming unit. In contrast, DCF-C and APA2-Lev have -1.58 and -4.16, respectively.

[0215] The antibiofilm activity of the multicomponent systems is shown as absolute colony-forming unit (CFU) counts, z.e., the direct, untransformed number of viable colony-forming units, for the individual components and for the combined formulation in Figures 3 A and 3B, respectively.

[0216] The antibiofilm activity of the multicomponent systems is presented as relative logarithmic CFU values, i.e., CFU data expressed on a log10scale relative to the controlcondition, for the individual components and for the combined formulation in Figures 4A and 4B, respectively.Table 6: Results of antibiofilm assay (individual compounds)Compound / CFU ACFU loglO Alogio GCombination (n = 3) (%) (CFU) (CFU) Untreated control 4,05E+07 4,17E+07DCFO 2,05E+06 3,55E+06 -94.93% 631 -1.29 C l,24E+08 l,87E+08 206.61% 8.09 0.49 APA1 1,11E+O8 7,62E+07 173.59% 8.04 0.44 APA2 8,18E+07 6,21E+07 102.13% 7.91 0.31Lev l,63E+06 7,12E+05 -95.97% 6.21 -1.39 Table 7. Results of antibiofilm assay (combinations)CFU ACFU loglO Alogio Combination(n = 3) (%) (CFU) (CFU) Untreated control 9.29E+07 2.92E+07 0.00% 7.97DCF-C 2.44E+06 3.00E+06 -97.37% 6.39 -1.58 APAl-Lev 1.25E+06 3.86E+05 -98.66% 6.10 -1.87 APA2-Lev 6.48E+03 6.56E+03 -99.99% 3.81 -4.16 DCF-C-APAl-Lev 3.00E+04 5.75E+03 -99.97% 4.48 -3.49DCF-C-APA2-Lev 5.92E+02 8.75E+02 -100.00% 2.77 -5.20

[0217] These results show the antibiofilm activity and synergistic effect of multicomponent DCF systems of the invention.Example 4: Biological evaluation - Antimicrobial activity (Pseudomonas)Materials and MethodsAntimicrobial components and multicomponent systems

[0218] The antimicrobial components and DCF-C-APAl-Lev were provided or prepared as described hereinabove (see Example 2).Screening of multicomponent systems for antimicrobial activity against A. baumannii

[0219] Sample solutions of individual components and multi-component systems were placed in a 96-well plate (50 pL each) and diluted with MilliQ water (1:2 dilution with 4% DMSO solution) to obtain seven concentrations, followed by adding a bacterial culture of Pseudomonas (150 pL). Bacterial growth monitoring was conducted at 37°C with periodic readings of optical density OD600over 24 hours.Minimum inhibitory concentration and Fractional inhibitory concentration index calculations

[0220] Minimum inhibitory concentration (MIC) was determined as the minimum concentration of antimicrobial agent or its combination, inhibiting bacterial growth during 24-hour monitoring time. The fractional inhibitory concentration index (FICI) was calculated as the ratio of the MIC of the component in the complex system to the MIC of an individual substance. To determine the overall effect of the combination, the sum of FIC for each system was compared with literature data (See Table 1 of Example 2).Results

[0221] Table 8 provides an overview of MIC and FICI values for the single components and the multicomponent system DCF-C-APAl-Lev.T ble 8. Summary table of the results of the analysis of antimicrobial activity (MICs and FICIs) (Pseudomonas)MICSystem Component Σ FICI Effect _ _DCF0 DCF0 500 - - C C 684 - - APA1 APA1 124 - - Lev Lev 56 - - DCF0 31DCF-C- C 22 Strong 0.23APAl-Lev APA1 8 synergyLev 4

[0222] These results show the antimicrobial activity and synergistic effect of multicomponent DCF systems against Pseudomonas based on MIC determination and FICI calculation, and confirm that pillar[5]arene-based multi-component DCF systems exhibit significant antimicrobial activity against Pseudomonas, particularly when combined with APA1 and Lev.Example 5: Biological evaluation multicomponent systems with Nb-Antibiofilm activityMaterials and Methods

[0223] Compounds, Nanobody protein, and combinations for antibiofilm assays. Six individual compounds and six combinations were chosen for antibiofilm assays: Dynamic polymer DCFO; Charged heads Aminoguanidine 3C; Pillararenes APA2; Antibiotic Levofloxacin Lev (see Example 2-3, and Nanobody (NB, also called herein “Non¬ functionalized NBO”, see Example 6). For subsequent studies of the antibiofilm activities of combinations, it was proposed to use solution concentrations equal to 4x the combinations' MIC (Table 9, MICs of the DCF-C and APAl-Lev combinations are reference).Table 9. The concentration of tested compounds and combinationsCompound / C(l), C (2),Comp 1 Comp 2 Combination jig / mL jig / mL NB NB 500DCF-C-APA2-Lev DCFO 500 C 344 DCF-C-APA2-Lev-NB DCFO 500 C 344APA2-Lev APA2 248 Lev 56T ble 9. (continued)Compound / Comp Comp C (3), Comp C (4), Comp C (5), Combination 1 3 jig / mL 4 jig / mL 5 jig / mL NB NB DCF-C-APA2- DCFO APA2 248 Lev 56LevDCF-C-APA2- DCF0 APA2 248 Lev 56 NB0 500Lev-NB

[0224] Antibiofilm assay setup. To allow the initial adhesion, a diluted suspension of bacteria (Acinetobacter baumannii strain AB5075) was added to a 96-well plate for 4 h. The medium was removed, and a mixture of fresh medium (low salt LB) and compound was added. The 96-well plate was incubated for another 20 h. The medium with compounds was removed, and bacteria were detached to perform a CFU count.Results

[0225] Table 10 present the results of the antibiofilm activity of multicomponent systems. The activity of the 2-component system APA2-Lev as shown in Example 3 was confirmed. Also, the complex dynamic system consisting of 4 components, such as DCF-C-APA2-Lev, shows enhanced antibiofilm properties compared to individual two- component systems tested in this experiment: 4-component combination DCF-C-APA2-Lev has a similar effect, the difference in log 10 of CFU counts compared to untreated sample is -4.78 with the result of 100 % reduction of the absolute value of colony forming unit. In contrast, APA2-Lev has -2.83 (99.85% reduction).

[0226] The complex system, composed of DCF0, Aminoguanidine C, Pillararene APA2, and Antibiotic Lev in combination with non-functionalized Nanobody NB0, showed excellent results on the biofilm inhibition: DCF-C-APA2-Lev has Alogio(CFU) = -4.78, and DCF-C-APA2-Lev-NB has Alogio(CFU) = -5.53. We observe a 16 % decrease in CFU count by introducing Nanobody into a complex dynamic system.Table 10. Results of antibiofilm assayErreur! Liaison incorrecte.Example 6: Production and characterization of Acinetobacter Omp25-specific Nanobody proteinsMaterials and Methods

[0227] Production of Acinetobacter specific Nbs. The Nb clones were transformed to E. coli WK6. Cells were grown in Terrific Broth (Duchefa Biochemie) and induced by IPTG for periplasmic expression and purification of the Nbs as described by Pardon et al. (2014,A general protocol for the generation of Nanobodies for structural biology. NatProtoc, 9 (3), 674-693), with each Nb sequence fused at its C-terminus to an affinity tag for detection purposes.

[0228] Microscopy. The classically used AB5075-VUB (CP070362), 3 Acinetobacter spp. isolated from environmental sources (A. calcoaceticus, A. junii and A. pittii), Klebsiella pneumoniae, and an E. coli S17 strain were used in the microscopy assays on living bacteria. The bacterial cultures were started from a single clone and were grown for 17 h at 37°C under agitation (165 rpm) in low salt broth (Luria-Bertani formulation, Duchefa Biochemie). The Brucella strains S. melitensis 16M, B. abortus 544 and B. abortus 2308) were grown overnight and then fixed with 4% paraformaldehyde for 1,5h before being used in the microscopy assays. Cells were collected by centrifugation (8000g) and normalized to ODeoo=3 (109CFU / ml) in phosphate buffered saline (PBS) for further steps. Labelling of the nanobody was done by incubation with molar 3 -fold of DyLight 650 NHS ester (ex:652 / em:672, Thermo Scientific) for 1 h, quenching of the reaction with 50 mM Tris buffer, followed by overnight dialysis in PBS. To allow binding, 100 pl of the labelled Nb (10 pM) was incubated with 100 pl of fixed or living bacterial cells (approximately 105CFU) for 30 min at 37°C, under agitation (165 rpm). To remove unbound Nb, the cells were centrifugated (8000g). Finally, the bacteria were spotted on an 1.5% agarose pad (Thermo Scientific Gene Frame). Microscopy images were acquired using a Leica DMi8 fluorescence microscope with a DFC7000 GT camera (Leica Microsystems CMS GmbH). The fluorescent images were acquired with a Leica FRAP450 and Y5 filter set. The raw data was processed by using ImageJ software where brightness was adjusted equally for all fluorescence micrographs within one experiment.ResultsNbH7 specifically binds the membrane of A. baumannii cells.

[0229] Fluorescence microscopy revealed that NbH7 (SEQ ID NO:4) bound the non¬ capsulated AB5075-VUB-itrA:: ISAbal3 (also referred to as AB5075C-) strain both in exponential and stationary phase (Figure 1). Furthermore, we additionally showed that NbH7 binds a variety of A. baumannii strains, including a multidrug-resistant (AB 180-VUB), an extensively drug-resistant (AB220-VUB) and a pandrug-resistant (AB3-VUB) strain and another classically used strain (ATCC17978-VUB), while the same experiment using E. coll cells did not result in binding of NbH7, indicating specificity of the Nb binding site.NbH7 specifically targets the Acinetobacter baumannii Outer-membrane-protein 25 (Omp25)

[0230] To demonstrate that the identified target of the Nb as revealed was indeed Acinetobacter Omp25, the AB5075C+Aomp25 and AB5075C-Aomp25 strains were generated. In Figure 2, we show that the strain lacking the omp25 gene did not result in binding of the NbH7 to the cells, providing the confirmation to conclude that Acinetobacter baumannii Omp25 is the antigen target of NbH7.

[0231] It is further envisaged herein that in additional to NbH7 (SEQ ID NO:4), further VHH or Nb polypeptides originating from the same VHH family, and / or which are optimized or humanized sequence variants of NbH7, with the same binding properties, may also serve as Nb components in the multicomponent system of the present invention.Example 7: Biological evaluation - Interaction between DCF-C and APAl-LevMaterials and Methods

[0232] The synergistic interaction between the dynamic constitutional framework polymer DCF-C and the APAl-Lev drug delivery system against Acinetobacter baumannii was quantified using a broth microdilution checkerboard assay. This method was also used to optimize the composition of the system DCF-C-APAl-Lev.

[0233] Assay Plate Preparation. The assay was performed in sterile 96-well U-bottom microtiter plates. Stock solutions of DCF-C and the APAl-Lev complex were prepared in sterile deionized water (or an appropriate buffer such as PBS, pH 7.4). A two-fold serial dilution of the DCF-C polymer was then performed vertically across the plate (e.g., rows B-H), while a two-fold serial dilution of the APAl-Lev system was performed horizontally (e.g., columns 2-8). This created a matrix of 49 unique concentration combinations. Control wells containing DCF-C only (column 1) and APAl-Lev only (row A) were included to determine their individual Minimum Inhibitory Concentrations (MICs). A positive control well (media and inoculum only) and a negative control well (media only) were also included.

[0234] Incubation and Analysis. Each well was inoculated with 150 pL of the prepared A. baumannii inoculum, bringing the final volume in each well to 200 pL. The plates were sealed and incubated at 37°C for 18-24 hours. Following incubation, the MIC was determined as the lowest concentration of an agent (alone or in combination) that completely inhibited visible bacterial growth.Results

[0235] Based on the Checkboard assay for the DCF-C-APAl-Lev combination, it was observed that there is a synergistic effect between the dynamic polymer DCF-C and the drug delivery complex APAl-Lev (Fig. 5). The shape of the graph showing the dependence of the combination’s minimum inhibitory concentration on different ratios is comparable to the typical synergistic relationship described in the scientific literature (Chait R., etal., “Antibiotic interactions that select against resistance'". Nature Vol. 446,pp. 668-671 (2007)). This method also enabled identification of the optimal combination with a molar ratio of DCF0: C: APAl: Lev = 1:1:0.05:0.1 by molar. This optimal combination yielded a Fractional Inhibition Concentration Index (FICI) of 0.318 (Clear synergistic effect between individual components) (see Table 11 and Fig. 5).Table 11. Calculation of the FICI for the optimized combination DCF-C-APAl-Lev Compound / MIC, jig / mL EFICI Combination DCF0 C APA1 LevDCF0 500C 10944APA1 124Lev 56DCF-C-APAl-Lev 31 22 16 7 0.318Example 8: Biological evaluation - Antimicrobial activity against (A. baumanniiMaterials and MethodsAntimicrobial components

[0236] The following substances were investigated for antimicrobial properties:- Dynamic constitutional framework consisting of Benzene-l,3,5-tricarbaldehyde and Poly-(ethyleneglycol)-bis(3-aminopropyl) terminated-PEG (1:1 molar ratio) - DCF0 core;DCF0- Pillar[5]arenes: Decaaminosubstituted Pillar[5]arene - AHPA2- Functional heads: Aminoguanidine hydrochloride - 3C;HCI NH2HN ^ NHI NH2C- Antibiotic: Levofloxacin - Lev.

[0237] Several combinations were evaluated, namely, DCF-C, AHPA2, AHPA2-Lev, and multi-component system DCF-C-APA2-Lev.Antimicrobial activity against A. baumannii

[0238] Sample solutions of individual components and multi-component systems were placed in a 96-well plate (50 pL each) and diluted with MilliQ water (1:2 dilution with 4% DMSO solution) to obtain seven concentrations, followed by adding a bacterial culture of A. baumannii (150 pL). Bacterial growth monitoring was conducted at 37°C with periodic readings of optical density OD600 over 24 hours.Minimum inhibitory concentration and Fractional inhibitory concentration index calculations

[0239] Minimum inhibitory concentration (MIC) was determined as the minimum concentration of antimicrobial agent or its combination, inhibiting bacterial growth during 24-hour monitoring time.

[0240] The fractional inhibitory concentration index (FICI) was calculated as the ratio of the MIC of the component in the complex system to the MIC of an individual substance. To determine the overall effect of the combination, the sum of FIC for each system was compared with literature data (Table 1).Results

[0241] Investigation of the antimicrobial activity of pillararene subcomponent AHPA2 alone and in combination with the antibiotic Lev (AHPA2-Lev), as well as with the dynamic polymer (DCF-C-AHPA2-Lev), showed a synergistic effect (Fig. 6 and Table 12) based on the calculation of the Fractional Inhibitory Concentration Index (FICI). For the AHPA2-Lev combination, the FICI was 0.5; for the DCF-C-AHPA2-Lev combination, it was 0.38. In both cases, these values are below or equal to 0.5, which according to the literature indicates a positive synergistic interaction between the components of the combination.Table 12. Minimal inhibition concentration MIC (pg / mL and mM) and Fractional inhibition concentration index FICI of DCF-C- AHPA2 -Lev combination.MICTested System Component FICI EFICI Comment pg / mL mMData from DCF0 DCF0 500 3.08 - - example 2 Data from C C >10944 >99 - - example 2 AHPA2 AHPA2 248 0.07 - - Data from Lev Lev 56 0.155 - - example 2Sa DCF0 125 0.77 0.25 Strong DCF-C 0.258C 86 0.78 0.008 synergy AHPA2 62 0.018 0.25AHPA2-Lev 0.5 Synergy Lev 14 0.039 0.25DCF0 63 0.39 0.126DCF-C- C 43 0.39 0.004 Strong 0.38AHPA2-Lev AHPA2 31 0.009 0.125 synergyLev 14 0.019 0.125

Claims

CLAIMS1. A molecular system comprising:(i) at least one dynamic constitutional framework (DCF) comprising:a plurality of cores, wherein each core is independently selected from phenyl, monocyclic heteroaryl, polycyclic aryl, and polycyclic heteroaryl; and a plurality of connectors, wherein each connector is a bifunctional linear homopolymer;wherein each of said connectors is covalently bound to two of said cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product;(ii) a plurality of functional heads, wherein each functional head is independently selected from amines, polyamines, hydrazides, and acyl-hydrazides;wherein each of said functional heads is covalently bound to one of said cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product;(iii) at least one pillararene; and(iv) at least one anti-infective agent;wherein each of said anti-infective agents is hosted in one of said pillararenes; andwherein each of said pillararenes is covalently bound to at least one of said cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product;wherein each of said cores is covalently bound to at least three groups selected from said connectors, said functional heads, and said pillararenes.

2. The molecular system according to claim 1, wherein each core is a trivalent phenyl, preferably an 1,3,5-subsituted phenyl.

3. The molecular system according to claim 1 or claim 2, wherein each connector is a polyethylene glycol (PEG) terminated by two (C1-C4) alkyls or polypropylene glycol (PPG) terminated by two (C1-C4) alkyls, preferably a polyethylene glycol (PEG) terminated by two propyls, more preferably a polyethylene glycol (PEG) terminated by two n-propyls.

4. The molecular system according to any one of claims 1 to 3, wherein said functional heads are selected from the following formulae (A) to (W)A (Mn= 800 Da)B (Mn= 2000 Da)wherein one of the -NH2 groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the functional head is covalently bound to one of the cores.

5. The molecular system according to claim 4, wherein said functional heads are selected from following formulae (B’) and (C’)NH2(B') HN NHNH2wherein one of the -NH2 groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the functional head is covalently bound to one of the cores; andwherein Mnis 2000 Da in the compound of formula (B’).

6. The molecular system according to any one of claims 1 to 5, wherein said pillararene is a pillar[5]arene, preferably selected from the following formulae (APAF), (APA2’), and (AHPA2’)(APAl1)(APA21)NH2NH3H-lJ.H(AHPA2’)wherein at least one of the -NH2 or -NH3+groups is replaced by a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product, by which the pillararene is covalently bound to at least one of the cores.

7. The molecular system according to any one of claims 1 to 6, wherein said anti-infective agent is an antibiotic agent; preferably said antibiotic agent is selected from levofloxacin (Lev), lincomycin (Lin) and cefalexin (Cef); more preferably said antibiotic agent is levofloxacin (Lev).

8. The molecular system according to any one of claims 1 to 7, wherein each reversible coupling product in said molecular system is independently selected from N=CH, ON=CH, S-S, S-CH2, Se-Se, B-O, B(O)-O, Si-O, Si-O-Si S(C=O)O, (C=O)O, (C=O)O, (C=O)O(C=O), O, S, NHC(NH2+)CH2, NHN=CH, C=C, C=C; and (C=O)NHN=CH.

9. The molecular system according to claim 8, wherein each reversible coupling product in said molecular system is N=CH.

10. The molecular system according to any one of claims 1 to 9, wherein said molecular system further comprises at least one antibody; preferably at least one immunoglobulin single variable domain (ISVD), such as a Variable Heavy domain of Heavy chain (VHH), or single domain antibody (sdAb) and / or preferably said antibody specifically binds to a cell surface of bacteria of the genus Acinetobacter, more preferably to a cell surface of bacteria of the species Acinetobacter baumannii, furthermore preferably to the protein Omp25 from Acinetobacter baumannii.

11. A pharmaceutical composition comprising a molecular system according to any one of claims 1 to 10 and at least one pharmaceutically acceptable carrier.

12. The molecular system according to any one of claims 1 to 10 or the pharmaceutical composition according to claim 11 for use as a medicament.

13. The molecular system according to any one of claims 1 to 10 or the pharmaceutical composition according to claim 11 for use in the treatment of an infectious disease, preferably a bacterial disease, more preferably a bacterial disease caused by Acinetobacter baumannii bacteria.

14. A process for manufacturing a molecular system according to any one of claims 1 to 10, wherein said process comprises the following steps:(a) a step of reacting a plurality of cores as defined in claim 1 with a plurality of connectors as defined in claim 1, thereby obtaining a base dynamic constitutional framework (base DCF); then(b) a step of reacting said base DCF with a plurality of precursors of a functional head as defined in claim 1, thereby obtaining a functionalised dynamic constitutional framework (DCF);(c) providing a host-guest complex comprising at least one pillararene as defined in claim 1 and at least one anti -infective agent as defined in claim 1, wherein each of said anti-infective agent(s) is hosted in one of said pillararene(s);(d) a step of reacting said functionalised dynamic constitutional framework (DCF) with said host-guest complex, thereby obtaining the molecular system according to any one of claims 1 to 9; and(e) optionally, contacting and / or reacting the molecular system with a plurality of immunoglobulin single variable domains (ISVDs), thereby obtaining the molecular system according to claim 10.

15. A hosting system comprising:(i) at least one dynamic constitutional framework (DCF) comprising a plurality of cores and a plurality of connectors, as defined in claim 1;(ii) a plurality of functional heads as defined in claim 1;wherein each of said functional heads is covalently bound to one of said cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product;(iii) at least one pillararene;wherein each of said pillararenes is covalently bound to at least one of said cores by means of a reversible coupling product or a (C1-C4) alkyl interrupted or terminated by at least one reversible coupling product;wherein each of said cores is covalently bound to at least three groups selected from said connectors, said functional heads, and said pillararenes andwherein each of said pillararenes does not host any anti-infective agent.

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