Cyclodextrins-bilirubin inclusion complexes

Abstract

The present invention relates to the production of cyclodextrin-bilirubin inclusion complexes. In particular, the invention relates to derivatives of cyclodextrins containing one or more groups designed to increase the capture of unbound bilirubin, which circulates freely in blood plasma, so that it does not create damage to the human body, in particular to the organism of newborns.

Description

[0001] Cyclodextrins-bilirubin inclusion complexes

[0002] Technical field

[0003] The present invention relates to the production of cyclodextrin- bilirubin inclusion complexes. In particular, the invention relates to derivatives of cyclodextrins containing one or more groups designed to increase the capture of unbound bilirubin, which circulates freely in blood plasma, so that it does not create damage to the human body, in particular to the organism of newborns.

[0004] Known art

[0005] Bilirubin (referred to in the text as BR) is a highly hydrophobic and water-insoluble derivative of tetrapyrrole. BR is the final product of the catabolism of heme. Specifically, in the human body haemoglobin is catabolized to produce heme, which is then reduced and oxidized to bilirubin. The bilirubin formed by this process first binds to the plasma albumin and then is transported to liver as an albumin-bilirubin complex.

[0006] A small fraction of serum bilirubin is not albumin-bound (referred to as albumin-unbound bilirubin or also called UCB) and can cross the blood brain barrier and damage brain tissues. Indeed, the alteration of the metabolic / transport pathways of bilirubin can lead to an increase in plasma concentration of albumin-unbound bilirubin and resulting hyperbilirubinaemia.

[0007] In some cases, newborns with severe hyperbilirubinaemia may develop acute bilirubin encephalopathy (ABE) and neurologic spectrum disorders of the kernicterus (KSD). Kernicterus is a disease that occurs in premature and term newborns, caused by high levels of unbound bilirubin (UCB; >20 mg). In this condition, UCB can cross the blood-brain barrier and accumulate in particular areas of the central nervous system, leading to permanent neurological dysfunction such as cerebral paralysis, permanent hearing loss and cognitive impairment or in some cases death. (Shapiro; 2003). Therefore, several national guidelines propose strategies to identify newborns at risk of hyperbilirubinaemia. About 50% of full-term newborns and 80% of preterm develop jaundice in the first week of life. Jaundice is also the most frequent cause of hospital readmission after postnatal discharge of newborns. Currently, the only effective method to prevent jaundice is to lower UCB levels by phototherapy (first choice therapy that converts bilirubin into a water-soluble isomer) or, in non-responsive cases, by exchange transfusion (EXT). In addition, phototherapy is required for 5-10% of full-term newborns and >50% of preterm newborns.

[0008] Jaundice usually appears 2 to 4 days after birth, and therefore many newborns require careful outpatient monitoring of bilirubin values after discharge (most healthy newborns are discharged on the third day).

[0009] A previous study by the inventors showed that unbound bilirubin is neurotoxic in both immature and mature rat organotypic hippocampus slices (model used to mimic in vitro the preterm and term brain). (Dani et al. Neonatology 2019;1 15:217-225). In these experimental conditions, human serum albumin (HSA), to which bilirubin is bound in plasma, is able to prevent UCB-induced neurotoxicity (in a molar ratio >1 HSA / unbound bilirubin) both in mature and immature hippocampus slices (Dani et al. Neonatology 2019;1 15:217-225). Unfortunately, the clinical use of HSA as a enhancer of the ability to bind plasma bilirubin, being a blood derivative with possible associated adverse effects. For this, a new therapeutic strategy for the treatment of jaundice and / or kernicterus is therefore a medical need.

[0010] Cyclodextrins (denoted by the acronym CD) are generally cup- shaped oligosaccharides, biocompatible and formed from α-(1 ,4)-bound glucose units. Their bilirubin encapsulation properties were also studied in a 1988 study (Kano K, et al. J. Chem. Soc. Chem. Commun. 1988).

[0011] In 2012, three types of cyclodextrins ( α-, β- and γ-cyclodextrin) were conjugated to a system of branched polyethylene imine (PEI), a highly reactive water-soluble polymer, to create new adsorbents for the capture of albumin-free bilirubin for use in hemoperfusion techniques (Wang Z, Colloids and Surfaces B: Biointerfaces 90 (2012) 248-253). The results of the study showed that the adsorbent containing β-cyclodextrin led to a better performance in capturing unbound bilirubin than the adsorbent containing α- and γ-cyclodextrin, even higher than bovine serum albumin. In addition, a molecular docking study showed that bilirubin has a stronger interaction with β-cyclodextrin in both 1 :1 and 1 :2 ratios.

[0012] However, the teachings of prior art present the following problems and disadvantages: phototherapy has acute (e.g.: dehydration, hypocalcemia, skin rash, bronze child syndrome, etc.) and late adverse effects (e.g.: oxidative stress, DNA damage, cancer).

[0013] On the other hand, the adverse effects of exchange transfusion are gas embolism, arrhythmias, acidosis, hyperkaliemia, hypocalcemia, hypoglycaemia, thrombocytopenia, necrotizing enterocolitis, etc.

[0014] The therapy with tin mesoporphyrin (Stanate®) prevents hyperbilirubinaemia by limiting heme oxygenase activity and has been shown to be effective in the treatment of newborns with haemolytic hyperbilirubinaemia (e.g., mother-child ABO and blood group Rh incompatibility, etc.). However, this drug is found to have numerous adverse effects.

[0015] Many pharmaceutical strategies, mostly based on hemoperfusion, have been proposed in recent decades as extracorporeal methods for the removal / capture of unbound bilirubin from plasma, as new and viable low-risk alternatives to phototherapy and blood transfusions (Nocentini et al., J Enzyme Inhib Med Chem 2022). However, the implementation of effective and blood-compatible systems for catching UCB remains a considerable challenge.

[0016] Platforms for the removal of UCB from blood by enzymatic destruction have also been proposed but unfortunately have not reached clinical application. In addition, oral administration of drugs such as Orlistat®, PEG or bile acid derivatives has been shown to induce reduction of bilirubinaemia by both the drugs alone and in combination with phototherapy. Again, no clinical application has been achieved. Unless specifically excluded in the detailed description below, what is described in this chapter shall be considered as an integral part of the detailed description.

[0017] Summary of the invention

[0018] It is an object of the present invention the production of derivatized cyclodextrins represented by the formula (I) explained below which are effective in counteracting the increase of free bilirubin (not bound to albumin) that occurs with the increase of bilirubinaemia (hyper- bilirubinaemia) and are used in pharmaceutical field for humans and especially for the newborn.

[0019] Another object of the present invention are the inclusion complexes of the derivatized cyclodextrins of formula (I) with bilirubin which are effective in counteracting the increase of free bilirubin (not albumin- bound) that occurs as the bilirubinaemia increases (hyper-bilirubinaemia) and are used in human pharmaceutical field, especially for the newborn.

[0020] Still another object of the present invention are pharmaceutical compositions comprising the derivatized cyclodextrins of formula (I) and their inclusion complexes to be used to counteract the increase in free bilirubin (not bound to albumin) that occurs as bilirubinaemia increases (hyper-bilirubinaemia). The compositions include the derivatized CD of formula (I) in a pharmaceutically acceptable vehicle.

[0021] Still another object of the present invention is the application of derivatized CD and its inclusion complexes as diagnostic agents to detect subjects with bilirubinaemia.

[0022] These and other aims, advantages and features of the present invention will be described more fully in a detailed description of the embodiments that follow, also the claims describe preferred variants of the invention, forming an integral part of this description.

[0023] Brief description of the figures

[0024] The further aims and advantages of this invention will be clear from the detailed description below of the examples of its realization (and its variants) and from the accompanying drawings, given purely for explanation and not for limitation, wherein:

[0025] Figure 1. Fig. 1. Example according to the invention of the structure of CD and 2-hydroxypropyl cyclodextrin;

[0026] Figure 2A-T. Some examples of the non-limiting derivatized cyclodextrins according to the invention; SG11-1087, SGAB1-1 , SGAB1-2, SGAB1-3, SG11-1176, SG11-1202, SG11-1219, SG11- 1099, SG11-1151 , AB4-537, AB4-532, AB4-536, AB4-534, AB4- 538, SGAB1-4, AB4-526, SGAB1-5, SGAB1-6, SGAB1-7, SGAB1-8;

[0027] Figure 3. Preliminary results of in vitro studies to test the effect of cyclodextrins α, β and y (panel A: qualitative analysis; panel B: quantitative analysis). After 14 days of culture, organotypic slices of rat hippocampus were treated with UCB (100 μM; black column) alone or in combination with α-cyclodextrin (α-CD), β-cyclodextrin (β-CD) and γ-cyclodextrin (γ-CD) at 1 :1 and 1 :2 ratios. Human albumin (HSA; endogenous UCB-ligand; white column) was used as positive control. At the end of the experiments, neuronal damage was measured by quantifying the intensity of fluorescence of propidium iodide (PI) in the CA1 region of the hippocampus (panel B);

[0028] Figure 4. Preliminary results of in vitro experiments on new derivatized cyclodextrins according to the invention. After 14 days of culture, organotypic slices of rat hippocampus were treated with UCB (100 μM; black column) alone or in combination with β-cyclodextrin (β- CD; grey column) and derivatized cyclodextrins (SG1 -1087, SGAB1 -1 , SGAB1 -2, SGAB1 -3, SG1 1 -1 176, SG11 -1202, SG11 - 1219, SG11 -1151 , SG11 -1099, AB4-526, SGAB1 -4, AB4-532, AB4-536, AB4-537, AB4-534, AB4-538, SGAB1 -5, SGAB1 -6, SGAB1 -7, SGAB1 -8) used in a 1 :2 ratio compared to UCB. In this experimental set β-cyclodextrin (β-CD) was used as a positive control. At the end of the experiments, neuronal damage was measured by quantifying the intensity of fluorescence of propidium iodide (PI) in the CA1 region of the hippocampus.

[0029] Definitions

[0030] In the present invention, the terms "derivative / s" and "conjugate / s" are to be considered as synonymous.

[0031] In the present invention, the term cyclodextrin (or CD) refers to a type of dextrins consisting of natural cyclic oligosaccharides formed by 6, 7 or 8 glucose monomers joined together with an a, 1 -4 glucoside bond and closed in a ring.

[0032] In the present invention the terms "cyclodextrin / s" or "CD" are intended to include both "cyclodextrins" and "hydroxypropyl- cyclodextrins", unless explicitly specified.

[0033] In the present invention, the term "inclusion complex" is to be considered as the adduct that cyclodextrin forms with free bilirubin, preventing its toxic action and facilitating its excretion.

[0034] In the context of the present invention, the term "carbonic anhydrase inhibitor(s)", or "CAI group", or more simply "CAI" refers to pharmacophores such as sulphonamides and their bioisosteres (such as sulfamates and sulfamides) capable of inhibiting Carbonic Anhydrase (CA) enzymes, binding directly to the zinc ion of the active site (Supuran C. T. Nature Rev. Drug Discov. 2008; 7:168-181 ). In particular, the inhibition of the CAs present at the level of red blood cells, where heme catabolism produces bilirubin, is sought, thus conveying the cyclodextrin- linker-CAI conjugate to this site, achieving a more effective capture of UCB.

[0035] In the present invention, the term "linker-CAI substituent group" or "linker-CAI" indicates the presence of spacer chemical groups of various kinds (linker), which optimally connect and distance the cyclodextrin portion from the carbon anhydrase inhibitor group (CAI). In addition, the choice of linker can also be used to increase the selectivity and inhibitory profile of these conjugates towards the isoforms of erythrocyte carbon anhydrase. In the present invention, the term "capture system / s" is considered synonymous with the term "inclusion complex".

[0036] In the present invention, the word "about" is used to indicate a numerical value equal to the value reported ± its standard deviation.

[0037] Detailed description

[0038] The invention relates to cyclodextrins derivatized with one or more substituents, the cyclodextrin and the substituent being linked by a spacer, hereinafter referred to as linker. These derivatized cyclodextrins are able to form effective inclusion complexes with bilirubin and will be used to form adducts with UCB for the removal / capture of UCB from the bloodstream.

[0039] The derivatized cyclodextrins are represented by the formula (I) explained below. Their ability to form inclusion complexes with bilirubin effectively allows them to be used in the prevention of jaundice in both term and preterm newborns, due to the presence of high amounts of albumin-free plasma bilirubin. Therefore, they are used in the pharmaceutical field to counteract all diseases related to pathological levels of unbound bilirubin in humans and especially for the newborn.

[0040] These derivatized cyclodextrins have the following general formula (I): wherein:

[0041] • CD is a cyclodextrin

[0042] • Linker and CAI are described below; • m=positive integer between 1 and 8.

[0043] • If there are more than one LINKER-CAI group, they may be the same or different.

[0044] The following general formulae also cover salified forms, optically active forms and racemic forms and mixtures thereof.

[0045] Preferably the derivatized cyclodextrin has the following general formula

[0046] (II): wherein:

[0047] Cyclodextrin is selected among:

[0048] • α- (with n=1 ), β- (with n=2) and γ- (with n=3) cyclodextrin

[0049] • R = H, hydroxypropyl

[0050] • and mixtures thereof;

[0051] Preferably the linker has the formula:

[0052] — Z— Q— W— wherein:

[0053] • Z is a group selected among nothing, CH2, O, NH, S, Se;

[0054] • Q is selected among spacers with one or more bidentate groups or combinations of these bidentate groups, selected among: alkylene, alkenylene, or alkynylene chains, linear, branched or cyclic, having 1 to 20 carbon atoms (C1-C20) or 1 to 12 carbon atoms (C1-C12), or 1 to 8 carbon atoms (C1-C8), or in some cases 1 to 6 carbon atoms (C1-C6); ether (-O-), thioether (-S-), ester (- COO-), thioester (-CSO-), amide or substituted amide, amine or substituted amine, urea or substituted urea, thiourea or substituted thiourea, carbamate or substituted carbamate, thiocarbamate or substituted thiocarbamate, dithiocarbamate or substituted dithiocarbamate, heterocycle or substituted heterocycle as for instance but not limited to piperidine, piperazine, aromatic or substituted aromatic group (such as but not limited to phenylene, naphthylene), heteroaromatic or substituted heteroaromatic (such as, but not limited to triazole, tetrazole, quinoline, thiadiazole, thiazole, oxazole), hydrazine or substituted hydrazine, hydrazone or substituted hydrazone, hydrazide or substituted hydrazide, hydroxylamine or substituted hydroxylamine, ketone or substituted ketone, diazene or substituted diazene, sulfonamide or substituted sulfonamide, acylsulphonamide or substituted acylsulphonamide, acylurea or substituted acylurea.

[0055] The term "substituted", as used here, refers to a functional group, in this case the spacer Q, wherein one or more hydrogen atoms contained within that functional group are substituted by one or more substituents.

[0056] The term "substituent", as used here, refers to a group which may be one or more atoms. Examples of substituents include, but are not limited to, a halogen (e.g., F, Cl, Br and I); an oxygen atom or an oxygen- containing group such as hydroxyl groups, alkoxy groups, aryloxy groups, arylalkyloxy groups, oxo (carbonyl) groups, carboxylic groups including carboxylic acids, carboxylates and carboxylated esters; a sulphur atom or a sulphur-containing group such as thiol groups, alkyl and aryl alkyl sulphide groups, sulphoxide groups, sulphone groups, sulfonamide groups; an atom of nitrogen or a group containing nitrogen, such as amines, azides, hydroxylamines, cyano, nitro groups, N-oxides, hydrazides and enamines. The term "alkyl chain", as used here, refers to linear or branched alkyl groups, substituted or unsubstituted, and cycloalkyl groups of 1 to about 20 carbon atoms (C1-C20), 1 to 12 carbon atoms (C1-C12), 1 to 8 carbon atoms (C1-C8), or in some cases 1 to 6 carbon atoms (C1-C6). Examples of linear alkyl groups include those with 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl and n- octyl. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl and 2,2- dimethylpropyl groups. As used here, the term "alkyl" includes n-alkyl groups, isoalkyl groups, as well as other branched forms of alkyl groups. Representative substituted alkyl groups may be substituted by one or more of the groups listed here, for instance amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy and halogen.

[0057] The term "alkenyl chain", as used here, refers to linear or branched alkyl groups, substituted or not substituted, from 2 to 20 carbon atoms (C2-C20), from 2 to 12 carbon atoms (C2-C12), from 2 to 8 carbon atoms (C2-C8), or in some cases 2 to 4 carbon atoms (C2-C4) and at least one carbon-carbon double bond. An alkynyl group is a fragment containing an open binding point on a carbon atom which would be formed if a hydrogen atom bound to a triply-bound carbon were removed from the molecule of an alkyne.

[0058] The term "cycloalkyl" as used here refers to cyclic alkyl groups, substituted or not substituted, such as but not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. In some cases, the cycloalkyl group may have from 3 to about 8-12 ring atoms, while in other cases the number of carbon atoms in the ring varies from 3 to 4, 5, 6 or 7. In some cases, cycloalkyl groups may have from 3 to 6 carbon atoms (C3-C6).

[0059] The term "acyl", as used in this context, refers to a group containing a carbonyl part wherein the group is bound by the carbonylic carbon atom. The carbon atom is also bound to another carbonylic carbon atom, which may be part of an alkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, heterocyclic, hetero-cycloalkyl, hetero-arylic, heteroaryl-alkyl or similar, both substituted or not substituted. In the special case wherein the carbonylic carbon atom is linked to a hydrogen, the group is a "formyl group", an acyl group as defined in this context. An acyl group may include 0 to about 12-40, 6 to 10, 1 to 5 or 2 to 5 additional carbon atoms bound to the carbonyl group. An example of an acyl group is the acrylic group. An acyl group may also include heteroatoms in the meaning of what has just been expressed. An example of an acyl group in the context of this text is the nicotinyl (pyridyl-3-carbonyl) group. Other examples include acetyl, benzyl, phenylacetyl, pyridyl acetyl, cinnamyl and acrylyl groups, and the like. When the group containing the carbon atom bound to the carbonyl carbon atom contains a halogen atom, the group is defined as " haloacyl group". An example is the trifluoroacetyl group.

[0060] The term "aryl" as used in this context refers to substituted or unsubstituted cyclic aromatic hydrocarbons which do not contain any heteroatoms in the cycle. Therefore, aryl groups include, but are not limited to, phenyl, azulenyl, hepthalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl. In some variants, aryl groups contain about 6 to 14 carbon atoms (C6-C14) or 6 to 10 carbon atoms (C6- C10) in the cyclic portions of the groups. Aryl groups may be non- substituted or substituted, as defined in this context. Representative substituted aryl groups may be mono-substituted or substituted more than once, for instance but not only phenyls, 2-, 3-, 4-, 5- or 6-substituted or naphthyl groups 2-8 substituted, which may be substituted with carbon-based or non-carbon-based groups as listed in this text.

[0061] The term "arylalkyl", as used in this context, refers to alkyl groups as defined above, wherein a bond with hydrogen or carbon of an alkyl group is substituted by a bond with an aryl group as defined above. Examples of aryl alkyl groups include groups such as benzyl and phenylethyl, as well as alkyl groups fused with an aryl group, such as 4- ethyl indanyl. Arylalkenyl groups are alkenyl groups as defined above, wherein a hydrogen or carbon bond of an alkyl group is substituted by a bond to an aryl group as defined above. The term "heterocycle" as used in this context refers to compounds with aromatic or non-aromatic rings, either substituted or not substituted, containing 3 or more members in the ring, one or more of which is a heteroatom such as, but not limited to, Boron (B), Nitrogen (N), Phosphorus (P), Oxygen (O) and Sulphur (S). Thus, a heterocyclic can be either a heterocycle or a heteroaryl cycle, or, if polycyclic, any combination thereof. In some cases, heterocyclic groups may have 3 to about 20 members in the ring, while in other cases they may have 3 to about 15 members in the ring. In some contexts, heterocyclic groups may include 3-8 carbon atoms (C3-C8), 3-6 carbon atoms (C3-C6) or 6-8 carbon atoms (C6-C8).

[0062] The term "heterocyclic group" includes species of fused rings which include fused aromatic and non-aromatic rings. Representative heterocyclic groups include, but are not limited to, pyrrolidinyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, chromanyl, indolinonyl, isoindolinonyl, furanyl, pyrrolidinyl, pyridinyl, pyrazinyl, pyrimidinyl, triazinyl, thiophenyl, tetrahydrofuran-2-yl, pyrrolyl, oxazolyl, oxadiazolyl, imidazolyl, triazolyl, tetrazolyl, benzoxazolinyl, benzothiazolinyl, and benzimidazolinyl.

[0063] The term "heterocycloalkyl" used here refers to alkyl groups as defined above, wherein a hydrogen or carbon atom of an alkyl group as defined above is substituted by a bond to a heterocyclic group as defined here. Representative examples of heterocycloalkyl groups include, but are not limited to, methylfuran-2-yl, methylfuran-3-yl, methylpyridin-3-yl, methyltetrahydrofuran-2-yl and propylindol-2-yl.

[0064] The term "heteroarylalkyl" used here refers to alkyl groups as defined above, wherein a hydrogen or carbon atom of an alkyl group is substituted by a bond to an heteroaryl group as defined here.

[0065] The term "alkoxy" as used here refers to an oxygen atom connected to an alkyl group, including a cycloalkyl group, as defined above. Examples of linear alkyl groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentyloxy, exyloxy and the like. Examples of branched alkoxy groups include, but are not limited to, isopropoxy, sec-butoxy, tert- butoxy, isopentyloxy, isohexyloxy and the like. Examples of cyclic alkoxy groups include, but are not limited to, cyclopropyloxy, cyclobutyloxy, cyclopentiloxy, cyclohexyloxy and the like. An alkoxy group may also include double or triple bonds and may contain heteroatoms. For instance, an allyl alkoxy group is an alkoxy group in the meaning given here. A methoxyethoxyl group is also an alkoxy group in the sense shown here, as well as a methylenedioxy group in a context where two adjacent atoms of a structure are substituted in this way.

[0066] The term "amine" used in this context refers to primary, secondary and tertiary amines which have, for instance, the formula N(group), wherein each group can be independently H or non-H, such as alkyl, aryl and similar groups. Amines include, but are not limited to R'R"N-, wherein each R' and R" may be H or may be selected independently, such as dialkylamines, diarylamines, alkylarylamines, heterocycloamines and the like. The term "amine" also includes ammonium ions used in this context.

[0067] W means aliphatic (alkyl, alkenyl, alkynyl, substituted or not), cyclic, aromatic or heteroaromatic functions which include but are not limited to: wherein:

[0068] • X, X1, X2and X3may be equal or different from each other and include but are not limited to, H, halogens, nitro groups, amine, hydroxyl groups, trifluoromethyl groups, methyl groups, ethyl groups, methoxy groups and acetamides;

[0069] • Y1is a C or N atom; • CAI is a carbon dioxide inhibiting group, for instance, but not limited to, it can be a sulfonamide, sulfamate, primary or secondary sulfamide as follows: wherein

[0070] R3may be H or methyl or ethyl.

[0071] The derivatized cyclodextrins of the invention can be obtained according to the following scheme.

[0072] The present invention aims to improve the capture of albumin-free bilirubin from plasma by using the derivatized cyclodextrins of general formula (I). These derivatized cyclodextrins are able to form host-guest inclusion complexes with UCB, encapsulating it and promoting / favouring its excretion.

[0073] In fact, advantageously, the derivatized cyclodextrins of the invention are readily eliminated through glomerular filtration.

[0074] CAI approach

[0075] The derivatized cyclodextrins with CAI-linker groups have been advantageously used where CAI (Figure 1 -4) is chosen among the Carbon Anhydrase Inhibitors (CAI) because about 80% of bilirubin is derived from the catabolism of red blood haemoglobin in the reticuloendothelial system.

[0076] Red blood cells (RBC) are rich in the ubiquitous human isoforms CA I and II to help convert carbon dioxide into proton and bicarbonate and vice versa, once the red blood cells reach the lungs.

[0077] It has been experimentally found that derivatized cyclodextrins wherein the linker is bound to CAI scaffolds maintain the ability of such simple macrocycles to encapsulate albumin-free bilirubin and also target red blood cells, where most of the bilirubin is produced, due to the high affinity interaction of CAI-linker- derivatized cyclodextrin with human CA (Table 1 ), improving overall the capture of the non-albumin bound bilirubin.

[0078] In addition, since the CAI of the clinically used diuretic agents, renal excretion of the derivatized cyclodextrins, and therefore of unbound bilirubin (encapsulated in cyclodextrin), is increased due to the increase in diuresis.

[0079] Treatment with derivatized cyclodextrins can be carried out in combination with phototherapy and / or hemoperfusion treatments especially in the most severe cases of hyperbilirubinaemia, i.e. secondary to haemolysis or when bilirubin exceeds 20 mg / dl in the bloodstream.

[0080] Treatment may also be proposed in less dangerous situations to decrease the serum level of albumin-free bilirubin and, thus, the neurotoxicity of bilirubin.

[0081] Therefore, one aim of the invention is to treat preterm and term newborns with severe unconjugated haemolytic hyperbilirubinaemia, who require phototherapy treatments and constitute the target population. In particular, the proposed derivatised cyclodextrins can also be used in case of mother-child incompatibility related to blood group Rh, ABO incompatibility, deficiency of the enzyme G6PD (glucose-6- phosphate dehydrogenase), ellipsocytosis, spherocytosis or when unconjugated bilirubin exceeds the dangerous serum threshold (e.g.: 20 mg / dL). When the production of bilirubin exceeds the liver’s capacity to dispose of it, the accumulation of bilirubin can become pathological (serum concentrations of bilirubin 20-25 mg / dl), and the newborn could suffer serious neurological damage if left untreated (bilirubin-induced neurological damage: BIND) and to determine a neurological syndrome called "kernicterus spectrum disorders". In these cases, bilirubin deposits in specific areas of the brain (e.g. basal nuclei, hippocampus) where it causes lesions that are associated with permanent outcomes such as spastic paralysis, choreoathetosis, neuro-sensory deafness and cognitive deficit (AAP 2004; Stevenson et al., 201 1 ). It is therefore important to keep serum bilirubin levels below threshold values detailed in specific nomograms for gestational age and postnatal age of the newborn, preferably when such unbound bilirubin exceeds the serum threshold of 20 mg / dL.

[0082] The onset of haemolytic diseases in newborns is estimated at 1 to 3 per 1000 pregnancies. Hyperbilirubinaemia may occur in patients with different comorbidities, such as prematurity (and its complications), hypoxic-ischemic encephalopathy (HIE) and / or infections that can damage the blood-brain barrier (BBB) further increasing the neurotoxicity of albumin-unbound bilirubin.

[0083] The therapeutic treatments implemented to date for the treatment of jaundice involve a variety of adverse effects (discussed above). In addition, the recently proposed approaches are based on hemoperfusion which is related to several blood side effects.

[0084] The derivatized cyclodextrins according to the invention are suitable for use in pharmaceuticals for all diseases related to pathological levels of human bilirubin, especially in neonatology, more particularly for preterm newborns.

[0085] The derivatized cyclodextrins of the present invention can be formulated as an injectable preparation. Formulations for parenteral use (both intramuscular and bolus) can be prepared using water for injectable preparations as a dispersing phase, isotonizing agents such as sodium chloride or glucose, phosphate buffer pH 7.4 and appropriate amount of derivatized cyclodextrin dispersed or solubilized in an appropriate vehicle. For single dose containers, the lyophilized formulation can be obtained together with the amount of water for injectable preparations useful for the reconstitution of the colloidal suspension. For multidose preparations, a suitable antimicrobial shall be introduced to be selected among sodium metabisulfite, phenol, cresol, methyl p-hydroxybenzoate, benzyl alcohol or similar.

[0086] Formulations may also contain one or more of the following: antibiotic agents, antimicrobial agents, antibacterial agents, antifungal agents, anti-inflammatory agents; preservatives and antioxidants. Can be added: pH regulating agents in the quantities necessary to obtain the desired pH, usually a physiological pH between 5 and 7, preferably between 6 and 7.

[0087] The pharmaceutical and / or cosmetic expert is able to prepare cosmetic and pharmaceutical formulations based on his experience and reading the technical information provided in the following description.

[0088] The derivatized cyclodextrins of the invention can be used in medicine as well as therapy for diagnosis, especially the predictive diagnosis of bilirubinaemia. More specifically, cyclodextrins can be used to monitor the progress of the disease. In this sense, the injectable formulation is the preferred one.

[0089] "EXAMPLES"

[0090] The systems for capturing unbound bilirubin proposed here are biocompatible, being based on the use of the derivatized cyclodextrins of formula (I), do not require intravenous hemotransfusion and have albumin-unbound bilirubin as a specific target compared to other existing therapies or removal methods, increasing capture performance and reducing complications. In particular, the following:

[0091] 1 ) the CAI-linker groups exposed on the outer edge of the derivatized cyclodextrins guide the conjugate system CAI-derivatized cyclodextrins to red blood cells where a high expression of human CA I and II is present, quickly collecting excess unbound bilirubin in the blood. This approach is particularly useful in the case of haemolytic jaundice, wherein red blood cell breakdown releases CA into plasma, attracting the conjugate CAI-derivatized cyclodextrins which captures unbound bilirubin.

[0092] It is the belief of the inventors that the implemented selectivity of derivatized cyclodextrins against UCB provides a more efficient treatment than HB, with exceptional results when other therapies have failed. In fact, derivatized cyclodextrins are an effective treatment for hyperbilirubinaemia in vitro and are proposed as drugs that can reduce the adverse effects of current therapies and the number and duration of hospital admissions for hyperbilirubinaemia.

[0093] As mentioned, in 2012 (ref. cited) the derivatized cyclodextrins were proposed as conjugates for a system of branched polyethylene imine, a highly reactive water-soluble polymer, to create new adsorbents for the capture of albumin-unbound bilirubin based on hemoperfusion. However, to the knowledge of the inventors, neither free derivatized cyclodextrin as a system for capturing albumin-unbound blood bilirubin has been proposed, nor an albumin-unbound bilirubin capture technology based on derivatized cyclodextrins such as the one proposed here.

[0094] In addition, it should be noted that especially when R of the linker- R is CAI all the mechanism of bilirubin excretion self-intensifies because CAI can increase diuresis and accelerates the excretion of cyclodextrin which in turn incorporates bilirubin.

[0095] The following examples are provided to illustrate the invention and are not to be considered as limiting its scope.

[0096] EXPERIMENTAL PART

[0097] A series of conjugated derivatized cyclodextrins has been synthesized and characterized. Examples are given in Fig. 3. The synthetic routes and purification methods are being optimized and implemented.

[0098] Scheme 1. General synthesis of derivatives SG1 1 -1087, SGAB1 -1 , SGAB1 -2.

[0099] Scheme 3. General Synthesis of Derivatives SG1 1 -1099, SG1 1 -1151.

[0100]

[0101] Scheme 4. General Synthesis of Derivatives AB4-526, AB4-532, AB4- 536, AB4-534, AB4-538, AB4-537, SGAB1 -4, SGAB1 -5.

[0102] Scheme 5. General Synthesis of Derivative SGAB1 -5. Experimental Part

[0103] Synthesis of CD-I, CD-N3, CD-NH(CH2)6OH.

[0104] Nal, NaN3or NH2(CH2)6OH (10 equiv.) was added to a solution of β-CD-OTs (0,1 g, 1 equiv.) in H2O or anhydrous DMF (5 mL) under stirring. The reaction mixture was then heated to 80°C. After monitoring by TLC, acetone (80 mL) was added, and the resulting suspension was filtered into the desired product. The intermediate was used for subsequent reactions without further purification.

[0105] General synthesis of SG11-1087, SGAB1-1 , SGAB1-2 (Scheme 1 ).

[0106] The appropriate sulfonamide A-C (5 equivalent) was added, together with catalytic DIPEA, to a solution of β-CD-SH (0.1 g, 1 equivalent) in anhydrous DMF (3 mL) under stirring and inert nitrogen atmosphere. The reaction mixture was then heated to 80°C. After monitoring by TLC, Et2O (50 mL) was added, and the resulting suspension was kept under stirring for another 5 minutes before decanting the supernatant. The resulting residue was then suspended in acetone (50 mL) and filtered under vacuum to obtain the desired product. The compound has been purified by Combiflash (H2O / ACN 5-40% v / v).

[0107] SG11-1087. Product SG11-1087 was obtained according to the general procedure described from sulfonamide A. White powder; Yield 42%; TLC in silica gel: Rf= 0.62 (H2O / ACN 20% v / v); δH(400 MHz,

[0108] DMSO-d6): 10.26 (s, 1 H, exchange with D2O, CONH), 7.78 (s, 4H, 4 x Ar-H), 7.27 (s, 2H, exchange with D2O, SO2NH2), 5.78 (m, 14H, exchange with D2O, 14 x OH), 4.88 (m, 7H, 7 x CH), 4.56 (m, 6H, exchange with D2O, 6 x OH), 3.67 (m, 28H, 28 x CH), 3.33 (m, 14H, 7 x CH2), 2.88 (m, 2H, CH2), 2.66 (t, J = 6.1 Hz, 2H, CH2); m / z (Positive ESI): calculated 1377.3881 [M+H]+, found 1377.3876 [M+H]+.

[0109] SGAB1-1. The product SGAB1-1 was obtained according to the general procedure described above from sulfonamide B. White powder; Yield 19%; TLC in silica gel: Rf= 0.62 (H2O / ACN 20% v / v); δH(400 MHz, DMSO-d6): 8.16 (t, J = 6.2 Hz, 1 H, exchange with D2O, CONH), 7.76 (d, J= 7.2 Hz, 2H, 2 x Ar-H), 7.45 (d, J= 7.2 Hz, 2H, 2 x Ar-H), 7.31 (s, 2H, exchange with D2O, SO2NH2), 5.77 (m, 14H, exchange with D2O, 14 x OH), 4.86 (m, 7H, 7 x CH), 4.57 (m, 6H, exchange with D2O, 6 x OH), 4.29 (d, J = 6.2 Hz, 2H, CH2), 3.64 (m, 28H, 28 x CH), 3.35 (m, 14H, 7 x CH2), 2.89 (m, 2H, CH2), 2.64 (t, J = 6.1 Hz, 2H, CH2); m / z (Positive ESI): calculated 1391.4038 [M+H]+, found 1391.4034 [M+H]+.

[0110] SGAB1-2. The product SGAB1-2 was obtained according to the general procedure described from sulfonamide C. White powder; yield 15%; TLC in silica gel: Rf= 0.61 (H2O / ACN 20% v / v); δH(400 MHz,

[0111] DMSO-d6): 8.02 (t, J = 6.4 Hz, 1 H, exchange with D2O, CONH), 7.78 (d, J= 7.5 Hz, 2H, 2 x Ar-H), 7.43 (d, J= 7.5 Hz, 2H, 2 x Ar-H), 7.31 (s, 2H, exchange with D2O, SO2NH2), 5.76 (m, 14H, exchange with D2O, 14 x OH), 4.85 (m, 7H, 7 x CH), 4.55 (m, 6H, exchange with D2O, 6 x OH), 3.63 (m, 28H, 28 x CH), 3.36 (m, 14H, 7 x CH2), 3.10 (m, 2H, CH2), 2.90 (m, 4H, 2 x CH2), 2.65 (m, 2H, CH2); m / z (Positive ESI): calculated 1405.4194 [M+H]+, found 1405.4198 [M+H]+.

[0112] General synthesis of SGAB1-3, SG11-1176, SGAB1-6, SGAB1-7, SG1 1-1202, SG11-1219 (Scheme 2).

[0113] The appropriate sulfonamide D-l (5 equivalent) was added, together with catalytic DIPEA, to a solution of β-CD-l (0.1 g, 1 equivalent) in anhydrous DMF (3 mL) under stirring and inert nitrogen atmosphere. The reaction mixture was then heated to 100°C. After monitoring by TLC, Et2O (50 mL) was added, and the resulting suspension was kept under stirring for another 5 minutes before decanting the supernatant. The resulting residue was then suspended in acetone (50 mL) and filtered under vacuum to obtain the desired product. The compound has been purified by Combiflash (H2O / ACN 5-40% v / v).

[0114] SGAB1-3. The product SGAB1-3 was obtained according to the general procedure described above from sulphonamide D. White powder; Yield 64%; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz, DMSO-d6): 7.79 (d, J = 7.5 Hz, 2H, 2 x Ar-H), 7.46 (d, J = 7.5 Hz, 2H, 2 x Ar-H), 7.34 (s, 2H, exchange with D2O, SO2NH2), 5.71 (m, 14H, exchange with D2O, 14 x OH), 4.88 (m, 7H, 7 x CH), 4.49 (m, 6H, exchange with D2O, 6 x OH), 3.61 (m, 30H, 28 x CH + CH2), 3.37 (m, 14H, 7 x CH2), 2.90 (m, 4H, 2 x CH2); m / z (Positive ESI): calculated 1303.4055 [M+H]+, found 1303.4052 [M+H]+.

[0115] SG11-1176. Product SG11-1176 was obtained according to the general procedure described from sulfonamide E. White powder; 70% yield; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz,

[0116] DMSO-d6): 7.79 (d, J = 7.5 Hz, 2H, 2 x Ar-H), 7.46 (d, J = 7.5 Hz, 2H, 2 x Ar-H), 7.34 (s, 2H, exchange with D2O, SO2NH2), 5.73 (m, 14H, exchange with D2O, 14 x OH), 4.87 (m, 7H, 7 x CH), 4.49 (m, 6H, exchange with D2O, 6 x OH), 3.65 (m, 28H, 28 x CH), 3.39 (m, 14H, 7 x CH2), 2.93 (m, 4H, 2 x CH2); m / z (Positive ESI): calculated 1317.4211 [M+H]+, found 1317.4214 [M+H]+.

[0117] SGAB1-6. The product SGAB1-6 was obtained according to the general procedure described from sulfonamide F. White powder; yield 22%; TLC in silica gel: Rf= 0.61 (H2O / ACN 20% v / v); δH(400 MHz,

[0118] DMSO-d6): 11.93 (s, 1 H, exchange with D2O, NHCONH), 8.25 (s, 2H, exchange with D2O, SO2NH2), 6.39 (t, 1 H, J= 6.7 Hz, exchange with D2O, NHCONH), 5.71 (m, 14H, exchange with D2O, 14 x OH), 4.85 (m, 7H, 7 x CH), 4.43 (m, 6H, exchange with D2O, 6 x OH), 3.64 (m, 28H, 28 x CH), 3.35 (m, 14H, 7 x CH2), 3.19 (m, 2H, CH2), 2.87 (t, J= 6.2 Hz, 2H, CH2); m / z (Positive ESI): calculated 1383.3848 [M+H]+, found 1383.3843 [M+H]+.

[0119] SGAB1-7. The product SGAB1-7 was obtained according to the general procedure described from the sulfonamide G. White powder; Yield 19%; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz, DMSO-d6): 12.05 (s, 1 H, exchange with D2O, NHCONH), 8.23 (s, 2H, exchange with D2O, SO2NH2), 6.48 (t, 1 H, J= 6.7 Hz, exchange with D2O, NHCONH), 5.72 (m, 14H, exchange with D2O, 14 x OH), 4.84 (m, 7H, 7 x CH), 4.48 (m, 6H, exchange with D2O, 6 x OH), 3.64 (m, 28H, 28 x CH), 3.35 (m, 14H, 7 x CH2), 3.11 (m, 2H, CH2), 1.39 (m, 8H, 4 x CH2); m / z (Positive ESI): calculated 1425.4317 [M+H]+, found 1425.4322 [M+H]+.

[0120] SG11-1202. Product SG11-1202 was obtained according to the general procedure described above from the sulfonamide H. White powder; Yield 38%; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH (400 MHz, DMSO-d6): 8.91 (s, 1 H, exchange with D2O, NHCONH), 7.68 (d, J = 8.2 Hz, 2H, 2 x Ar-H), 7.54 (d, J = 8.2 Hz, 2H, 2 x Ar-H), 7.14 (s, 2H, exchange with D2O, SO2NH2), 6.30 (t, 1 H, J= 5.5 Hz, exchange with D2O, NHCONH), 5.70 (m, 14H, exchange with D2O, 14 x OH), 4.88 (m, 7H, 7 x CH), 4.45 (m, 6H, exchange with D2O, 6 x OH), 3.66 (m, 28H, 28 x CH), 3.36 (m, 14H, 7 x CH2), 3.18 (m, 2H, CH2), 2.86 (t, J= 6.2 Hz, 2H, CH2); m / z (Positive ESI): calculated 1375.4379 [M+H]+, found 1375.4376 [M+H]+.

[0121] SG11-1219. Product SG11-1219 was obtained according to the general procedure described from sulfonamide I. White powder; 34% yield; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz,

[0122] DMSO-d6): 8.00 (s, 1 H, exchange with D2O, NHCONH), 7.66 (d, J = 8.1 Hz, 2H, 2 x Ar-H), 7.54 (d, J = 8.1 Hz, 2H, 2 x Ar-H), 7.13 (s, 2H, exchange with D2O, SO2NH2), 6.43 (t, 1 H, J= 5.6 Hz, exchange with D2O, NHCONH), 5.70 (m, 14H, exchange with D2O, 14 x OH), 4.86 (m, 7H, 7 x CH), 4.50 (m, 6H, exchange with D2O, 6 x OH), 3.62 (m, 28H, 28 x CH), 3.37 (m, 14H, 7 x CH2), 3.09 (m, 2H, CH2), 1.40 (m, 8H, 4 x CH2); m / z (Positive ESI): calculated 1417.4848 [M+H]+, found 1417.4845 [M+H]+.

[0123] General synthesis of SG11-1099, SG11-1151 (Scheme 3).

[0124] The appropriate sulfonamide J or K (5 equiv.) was added, together with sodium ascorbate (10 equiv.) and CuSO4·5H2O (5 equiv.), to a solution of β-CD-N3(0,1 g, 1 equiv.) in anhydrous DMF (3 mL) under stirring and inert nitrogen atmosphere. The reaction mixture was then heated to 50°C. After monitoring by TLC, a 1 :2 mixture of H2O / MeOH (10 mL) was added and the resulting suspension filtered on Celite. The resulting solution was evaporated under vacuum to obtain the desired product. The compound has been purified by Combiflash (H2O / ACN 5- 40% v / v).

[0125] SG11-1099. Product SG11-1099 was obtained according to the general procedure described from the sulfonamide J. White powder; Yield 27%; TLC in silica gel: Rf= 0.61 (H2O / ACN 20% v / v); δH( 400 MHz,

[0126] DMSO-d6): 8.25 (s, 1 H, Ar-H), 7.81 (d, J= 7.2 Hz, 2H, 2 x Ar-H), 7.25 (m, 4H, exchange with D2O, SO2NH2+ 2 x Ar-H), 5.75 (m, 14H, exchange with D2O, 14 x OH), 5.22 (s, 2H, CH2), 5.09 (s, 1 H, CH), 4.87 (m, 6H, 6 x CH), 4.55 (m, 6H, exchange with D2O, 6 x OH), 4.36 (s, 1 H, CH), 4.04 (s, 1 H, CH), 3.67 (m, 26H, 27 x CH), 3.41 (m, 14H, 7 x CH2); m / z (Positive ESI): calculated 1371.4066 [M+H]+, found 1371.4060 [M+H]+.

[0127] SG11-1151. Product SG11-1151 was obtained according to the general procedure described from the sulfonamide K. White powder; 71% yield; TLC in silica gel: Rr = 0.60 (H2O / ACN 20% v / v); δH(400 MHz, DMSO-d6): 9.18 (s, 1 H, exchange with D2O, CONH), 8.04 (d, J= 7.6 Hz, 2H, 2 x Ar-H), 7.99 (s, 1 H, Ar-H), 7.93 (d, J= 7.6 Hz, 2H, 2 x Ar-H), 7.52 (s, 2H, exchange with D2O, SO2NH2), 5.80 (m, 14H, exchange with D2O, 14 x OH), 5.08 (s, 1 H, CH), 4.87 (m, 6H, 6 x CH), 4.55 (m, 6H, exchange with D2O, 6 x OH), 4.41 (t, J = 6.3 Hz, 1 H, CH), 4.01 (t, J = 6.3 Hz, 2H, CH2), 3.67 (m, 27H, 27 x CH), 3.40 (m, 14H, 7 x CH2); m / z (Positive ESI): calculated 1398.4175 [M+H]+, found 1398.4172 [M+H]+.

[0128] General synthesis of AB4-537, AB4-532, AB4-536, AB4-534, AB4- 538, SGAB1-4, SGAB1-8 (Scheme 4).

[0129] The appropriate L-R (2 equiv.) sulfonamide was added, together with catalytic DIPEA, to a solution of CD-NH(CH2)6NH2(0,1 g, 1 equiv.) in anhydrous DMF (3 mL) under stirring and inert nitrogen atmosphere, cooled to 0°C. The reaction mixture was then left at room temperature. After TLC monitoring, Et2O (50 mL) was added, and the resulting suspension was kept under stirring for additional 5 minutes before decanting the supernatant. The resulting residue was then suspended in acetone (50 mL) and filtered under vacuum to obtain the desired product. The compound has been purified by Combiflash (H2O / ACN 5-40% v / v).

[0130] AB4-537. The product AB4-537 was obtained according to the general procedure reported from the sulfonamide L. White powder; Yield 28%; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz,

[0131] DMSO-d6): 12.01 (s, 1 H, exchange with D2O, NHCONH), 8.21 (s, 2H, exchange with D2O, SO2NH2), 6.42 (t, 1 H, J= 6.7 Hz, exchange with D2O, NHCONH), 5.75 (m, 14H, exchange with D2O, 14 x OH), 4.86 (m, 7H, 7 x CH), 4.49 (m, 6H, exchange with D2O, 6 x OH), 3.65 (m, 28H, 28 x CH), 3.37 (m, 14H, 7 x CH2), 3.09 (m, 2H, CH2), 1.36 (m, 10H, 5 x CH2); m / z (Positive ESI): calculated 1439.4474 [M+H]+, found 1439.4471 [M+H]+.

[0132] AB4-532. The product AB4-532 was obtained according to the general procedure reported from the sulfonamide M. White powder; Yield 33%; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz, DMSO-d6): 8.84 (s, 1 H, exchange with D2O, NHCONH), 7.68 (d, J = 8.1 Hz, 2H, 2 x Ar-H), 7.55 (d, J = 8.1 Hz, 2H, 2 x Ar-H), 7.17 (s, 2H, exchange with D2O, SO2NH2), 6.32 (t, 1 H, J= 5.7 Hz, exchange with D2O, NHCONH), 5.74 (m, 14H, exchange with D2O, 14 x OH), 4.86 (m, 7H, 7 x CH), 4.49 (m, 6H, exchange with D2O, 6 x OH), 3.61 (m, 28H, 28 x CH), 3.39 (m, 14H, 7 x CH2), 3.10 (m, 2H, CH2), 1.37 (m, 10H, 5 x CH2); m / z (Positive ESI): calculated 1431.5005 [M+H]+, found 1431.5008 [M+H]+.

[0133] AB4-536. The product AB4-536 was obtained according to the general procedure reported from the sulfonamide N. White powder; Yield 29%; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz,

[0134] DMSO-d6): 8.78 (s, 1 H, exchange with D2O, NHCONH), 8.00 (s, 1 H, Ar- H), 7.53 (d, J = 8.3 Hz, 1 H, Ar-H), 7.34 (t, J = 8.3 Hz, 1 H, Ar-H), 7.35 (d, J= 8.3 Hz, 1 H, Ar-H), 7.31 (s, 2H, exchange with D2O, SO2NH2), 6.23 (t, 1 H, J= 6.4 Hz, exchange with D2O, NHCONH), 5.77 (m, 14H, exchange with D2O, 14 x OH), 4.87 (m, 7H, 7 x CH), 4.48 (m, 6H, exchange with D2O, 6 x OH), 3.64 (m, 28H, 28 x CH), 3.38 (m, 14H, 7 x CH2), 3.1 1 (m, 2H, CH2), 1.37 (m, 10H, 5 x CH2); m / z (Positive ESI): calculated 1431.5005 [M+H]+, found 1431.5008 [M+H]+.

[0135] AB4-534. The product AB4-534 was obtained according to the general procedure described above from the sulfonamide O. White powder; yield 25%; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz, DMSO-d6): 7.78 (d, J = 7.4 Hz, 2H, 2 x Ar-H), 7.43 (d, J = 7.4 Hz, 2H, 2 x Ar-H), 7.31 (s, 2H, exchange with D2O, SO2NH2), 7.41 (t, 1 H, J = 6.1 Hz, exchange with D2O, NHCONH), 6.01 (t, 1 H, J = 6.1 Hz, exchange with D2O, NHCONH), 5.76 (m, 14H, exchange with D2O, 14 x OH), 4.86 (m, 7H, 7 x CH), 4.48 (m, 6H, exchange with D2O, 6 x OH), 4.28 (d, 2H, J = 6.1 Hz, CH2), 3.66 (m, 28H, 28 x CH), 3.39 (m, 14H, 7 x CH2), 3.01 (m, 2H, CH2), 1.30 (m, 10H, 5 x CH2); m / z (Positive ESI): calculated 1445.5161 [M+H]+, found 1445.5166 [M+H]+.

[0136] AB4-538. The product AB4-538 was obtained according to the general procedure reported from the sulfonamide P. White powder; Yield 27%; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz,

[0137] DMSO-d6): 7.76 (d, J = 7.8 Hz, 2H, 2 x Ar-H), 7.41 (d, J = 7.8 Hz, 2H, 2 x Ar-H), 7.30 (s, 2H, exchange with D2O, SO2NH2), 6.05 (t, 1 H, J = 6.3 Hz, exchange with D2O, NHCONH), 5.95 (t, 1 H, J = 6.0 Hz, exchange with D2O, NHCONH), 5.75 (m, 14H, exchange with D2O, 14 x OH), 4.86 (m, 7H, 7 x CH), 4.48 (m, 6H, exchange with D2O, 6 x OH), 3.64 (m, 28H, 28 x CH), 3.38 (m, 14H, 7 x CH2), 3.15 (m, 2H, CH2), 2.98 (m, 2H, CH2), 1 .31 (m, 10H, 5 x CH2); m / z (Positive ESI): calculated 1459.5318 [M+H]+, found 1459.5322 [M+H]+.

[0138] SGAB1-4. The product SGAB1-4 was obtained according to the general procedure described above from the sulfonamide Q. White powder; Yield 31 %; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz, DMSO-d6): (400 MHz, DMSO-d6): 9.19 (s, 1 H, exchange with D2O, NHCSNH), 7.72 (d, J = 7.9 Hz, 2H, 2 x Ar-H), 7.60 (d, J = 7.9 Hz, 2H, 2 x Ar-H), 7.18 (s, 2H, exchange with D2O, SO2NH2), 6.81 (t, 1 H, J= 5.7 Hz, exchange with D2O, NHCSNH), 5.73 (m, 14H, exchange with D2O, 14 x OH), 4.85 (m, 7H, 7 x CH), 4.47 (m, 6H, exchange with D2O, 6 x OH), 3.59 (m, 28H, 28 x CH), 3.37 (m, 14H, 7 x CH2), 3.1 1 (m, 2H, CH2), 1.38 (m, 10H, 5 x CH2); m / z (Positive ESI): calculated 1447.4776 [M+H]+, found 1447.4779 [M+H]+.

[0139] SGAB1-8. The product SGAB1-8 was obtained according to the general procedure described from the sulfonamide R. White powder; yield 18%; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz, DMSO-d6): 9.1 1 (s, 1 H, exchange with D2O, NHCSNH), 8.17 (s, 1 H, Ar- H), 7.66 (d, J = 8.1 Hz, 1 H, Ar-H), 7.41 (t, J = 8.1 Hz, 1 H, Ar-H), 7.42 (s, 2H, exchange with D2O, SO2NH2), 7.30 (d, J= 8.1 Hz, 1 H, Ar-H), 6.65 (t, 1 H, J = 6.7 Hz, exchange with D2O, NHCSNH), 5.73 (m, 14H, exchange with D2O, 14 x OH), 4.85 (m, 7H, 7 x CH), 4.42 (m, 6H, exchange with D2O, 6 x OH), 3.66 (m, 28H, 28 x CH), 3.35 (m, 14H, 7 x CH2), 3.09 (m, 2H, CH2), 1.38 (m, 10H, 5 x CH2); m / z (Positive ESI): calculated 1447.4776 [M+H]+, found 1447.4779 [M+H]+.

[0140] Synthesis of AB4-526 (Scheme 4).

[0141] The appropriate sulfonamide S (2 equiv.) was added, together with NaHCO3(2 equiv.), to a solution of CD-NH(CH2)6NH2(0,1 g, 1 equiv.) in anhydrous DMF (3 mL) under stirring and inert nitrogen atmosphere, cooled to 0°C. The reaction mixture was then left at room temperature. After TLC monitoring, Et2O (50 mL) was added and the resulting suspension was kept under stirring for additional 5 minutes before decanting the supernatant. The resulting residue was then suspended in acetone (50 mL) and filtered under vacuum to obtain the desired product AB4-526. The compound has been purified by Combiflash (H2O / ACN 5- 40% v / v).

[0142] White powder; 49% yield; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v; δH(400 MHZ, DMSO-d6): 8.27 (s, 1 H, exchange with D2O, CONH), 8.02 (d, J= 8.2 Hz, 2H, 2 x Ar-H), 7.79 (d, J= 8.2 Hz, 2H, 2 x Ar-H), 7.51 (s, 2H, exchange with D2O, SO2NH2), 5.83 (m, 14H, exchange with D2O, 14 x OH), 4.86 (m, 7H, 7 x CH), 4.52 (m, 6H, exchange with D2O, 6 x OH), 3.65 (m, 28H, 28 x CH), 3.40 (m, 14H, 7 x CH2), 3.08 (m, 2H, CH2), 1 .40 (m, 10H, 5 x CH2); m / z (Positive ESI): calculated 1416.4896 [M+H]+, found 1416.4892 [M+H]+.

[0143] Synthesis of SGAB1-5 (Scheme 5).

[0144] The appropriate sulfonamide T (2 equiv.) was added, together with catalytic DMAP and EDC-HCI (2 equiv.), to a solution of CD-NH(CH2)6OH (0,1 g, 1 equiv.) in anhydrous DMF (3 mL) under stirring and inert nitrogen atmosphere, cooled to 0°C. The reaction mixture was then left at room temperature. After TLC monitoring, Et2O (50 mL) was added and the resulting suspension was kept under stirring for additional 5 minutes before decanting the supernatant. The resulting residue was then suspended in acetone (50 mL) and filtered under vacuum to obtain the desired product SGAB1-5. The compound has been purified by Combiflash (H2O / ACN 5-40% v / v). White powder; Yield 11%; TLC in silica gel: Rf= 0.60 (H2O / ACN 20% v / v); δH(400 MHz, DMSO-d6): 8.04 (d, J = 8.0 Hz, 2H, 2 x Ar-H), 7.82 (d, J = 8.0 Hz, 2H, 2 x Ar-H), 7.50 (s, 2H, exchange with D2O, SO2NH2), 5.82 (m, 14H, exchange with D2O, 14 x OH), 4.85 (m, 7H, 7 x CH), 4.51 (m, 6H, exchange with D2O, 6 x OH), 3.63 (m, 30H, 28 x CH + CH2), 3.38 (m, 14H, 7 x CH2), 1 .41 (m, 10H, 5 x CH2); m / z (Positive ESI): calculated 1417.4736 [M+H]+, found 1417.4740 [M+H]+.

[0145] Table 1. Inhibition data of the target isoforms of carbon anhydrase (CA)

[0146] I and II with the cyclodextrins proposed here. Acetazolamide (AAZ) was used as a standard. a. Average of 3 separate measurements with a Stopped Flow method (errors in the range 5-10 % of the tabulated values)

[0147] The cyclodextrin derivatives shown here have been tested in vitro for their ability to bind CA I and II present in erythrocytes. Inhibition data are shown in Table 1 and were through a Stopped Flow assay of CA catalysed CO2hydration activity. All compounds have shown inhibitory and therefore binding capacity to target CAs.

[0148] Simple and derivatized cyclodextrins were tested in an in vitro model of hyperbilirubinaemia on rat hippocampal organotipes (Dani et al. Neonatology 2019;115:217-225, Dani C, et al. Front. Pediatr. 2021 ;9:659477), obtained from 7-9 days Wistar rats, as reported in (Gerace E, et al. Methods Mol Biol. 2012;846:343-54). Hyperbilirubinaemia was reproduced in vitro by exposing hippocampal organotypic slices at 48 hours incubation with UCB (albumin-unbound bilirubin, 100 μM). In these experimental conditions, UCB induces selective damage in the CA1 region of the hippocampus (Dani C, et al. Neonatology 2019;115:217-225), which is quantified by measuring the fluorescence intensity of propidium iodide (PI).

[0149] Human albumin (HSA) has been used as a positive control, since it is an endogenous scavenging agent for UCB.

[0150] The preliminary results obtained from these experiments are given below as qualitative (Fig. 3A) and quantitative (Fig. 3B, 4) analyses obtained by quantifying the intensity of fluorescence of propidium iodide (PI) in the CA1 region of the hippocampus as a measure of neuronal damage.

[0151] The results obtained show that β-CD is able to reduce neurotoxicity induced by UCB significantly when their ratio is 1 :2 (Fig. 3). This data confirm a previous study wherein it was shown that the adsorbent that included β-CD had a better performance in capturing unbound bilirubin than the adsorbent that included α- and γ-CD. The derivatized cyclodextrins according to the invention (SG1 -1087, SGAB1 -1 , SGAB1 - 2, SGAB1 -3, SG11 -1 176, SG11 -1202, SG11-1219, SG1 1 -1 151 , SG11 - 1099, AB4-526, SGAB1 -4, AB4-532, AB4-536, AB4-537, AB4-534, AB4- 538, SGAB1 -5, SGAB1 -6, SGAB1 -7, SGAB1 -8) show a damage protection activity from UCB comparable to the β-CD (Fig. 4), and having the CA inhibitory function can specifically target red blood cells at blood level to incorporate the released UCB The examples described above may be made in other ways, without however leaving the scope of protection of the present invention and include all the realizations equivalent to the content of the claims for a technician in the field.

[0152] From the above description, the technician is able to realize the object of the invention without having to introduce further details.

Claims

CLAIMS1. A denvatized cyclodextrin having the following general formula (I):wherein:CD is a cyclodextrin of formulawherein: α- (with n=1 ), β- (with n=2) and γ- (with n=3) cyclodextrinR = H, hydroxypropyl and mixtures thereof; and wherein preferably R = H, -CH2(CHOH)CH3. m = positive integer number between 1 and 8LINKER has the formula Z Q WZ is a group selected among nothing, CH2, O, NH, S, Se;Q is selected among spacers with one or more bidentate groups or combinations of these bidentate groups, selected among: alkyl-, alkenyl-, or alkynyl-, linear, branched or cyclic chains having 1 to 20 carbon atoms (C1-C20) or 1 to 12 carbon atoms (C1-C12), or 1 to 8 carbon atoms (C1- C8), or in some cases 1 to 6 carbon atoms (C1-C6); ether (-O-), thioether (-S-), ester (-COO-), thioester (-CSO-), amide or substituted amide, amine or substituted amine, urea or substituted urea, thiourea or substituted thiourea, carbamate or substituted carbamate, thiocarbamate or substituted thiocarbamate, dithiocarbamate or substituted dithiocarbamate, heterocycle or substituted heterocycle, such as, for instance, piperidine, piperazine, aromatic or substituted aromatic group, such as phenylene, naphthylene, heteroaromatic or substituted heteroaromatic, such as, for instance, triazole, tetrazole, quinoline, thiadiazol, thiazole, oxazole, hydrazine or substituted hydrazine, hydrazone or substituted hydrazone, hydrazide or substituted hydrazide, hydroxylamine or substituted hydroxylamine, ketone or substituted ketone, diazene or substituted diazene, sulfonamide or substituted sulfonamide, acylsulphonamide or substituted acylsulphonamide, acyl urea or substituted acyl urea;W is selected among an alkyl, alkenyl or alkynyl chain, linear, branched or cyclic, with 1 to 20 carbon atoms (C1-C20), or 1 to 12 carbon atoms (C1- C12), or 1 to 8 carbon atoms (C1-C8) or, in some cases, 1 to 6 carbon atoms (C1-C6), aromatic or heteroaromatic groups, thiol cyclic groups selected among:wherein:X, X1, X2and X3, which are independently equal or different from each other, are: H, halogen, nitro group, amine, hydroxyl, trifluoromethyl, methyl, ethyl, methoxy, acetamide;Y1is a C or N atom; CAI is a group of carbon anhydrase inhibitors selected among: sulfonamide, sulfamate, primary or secondary sulfamide, preferably with the formula:wherein: R3is H or methyl or ethyl in the general formula (I) being comprised the salified forms, optically active forms and racemic forms and mixtures thereof.

2. The derivatized cyclodextrin according to the preceding claim wherein the substituents Z, Q, W and CAI are selected among those indicated in Table 1 :Table 13. Compositions comprising at least one derivatized cyclodextrin according to anyone of claims 1 -2 and one or more pharmaceutically acceptable components.

4. The compositions according to the preceding claim further comprising one or more: preservatives, antioxidants, pH regulators.

5. The compositions according to anyone of claims 3-4 further comprising one or more drugs selected among: antibiotics, antimicrobials, antibacterials, antifungal, anti-inflammatory.

6. Compositions according to anyone of claims 3-5 in the form of tablets, capsules, pills, injectable formulations, freeze-dried forms which may be reconstituted.

7. Derivatized cyclodextrins according to anyone of claims 1 -2 or compositions according to anyone of claims 3-6 for use as a medicament.

8. Derivatized cyclodextrins or compositions according to the previous claim for use in the treatment of conditions related to increased plasma concentration of albumin-free bilirubin (UCB) and resulting hyperbilirubinaemia.

9. Derivatized cyclodextrins or compositions according to the previous claim for use in the treatment of hyperbilirubinaemia in humans, particularly in term and preterm newborns.

10. Derivatized cyclodextrins or compositions according to the previous claim for use in the treatment of UCB-induced neurotoxicity, acute bilirubin encephalopathy and neurologic disorders of the kernicterus spectrum, in particular cerebral palsy, permanent hearing loss and cognitive impairment, haemolytic diseases of the newborn, mother-child incompatibility related to Rh blood group, ABO incompatibility, deficiency of the enzyme G6PD (glucose-6-phosphate dehydrogenase), ellipsocytosis, spherocytosis, or in cases where the serum threshold of albumin-unbound bilirubin exceeds 20 mg / dL.

11. Derivatized cyclodextrins or compositions for use according to anyone of claims 7-10 wherein the administration is by oral, parenteral intramuscular and bolus, intravenous, intracutaneous route.

12. A single or multi-dose preparation kit comprising one or more derivatized cyclodextrins according to anyone of claims 1 -2 in liquid form or lyophilized form to be reconstituted, one or more containers, water or solution for injectable preparations, one or more antimicrobial agents, instructions for use.

13. The inclusion complex between the derivatized cyclodextrin according to anyone of claims 1 -2 and serum albumin-unbound bilirubin.

14. A method for the treatment of diseases related to hyperbilirubinaemia in humans, especially in neonatology, more particularly in preterm newborns, wherein an effective dose of derivatized cyclodextrins according to anyone of claims 1 -2 or of compositions according to anyone of claims 3-6 is administered to a subject who needs it.

15. The method according to the previous claim wherein levels of freely circulating unbound bilirubin in blood exceed the serum threshold of 20 mg / dL.

16. The method according to claims 14-15 wherein hyperbilirubinaemia is associated with: prematurity and its complications, hypoxic-ischemic encephalopathy (HIE), infections that damage the blood-brain barrier (BBB), neurological damage (BIND), kernicterus spectrum disorders, lesions associated with spastic paralysis, choreoathetosis, neuro-sensory deafness and cognitive impairment.

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