Ionizable pillararene anticoagulant

Abstract

Provided are methods for inhibiting coagulation in a subject in need thereof using ionizable pillararene compounds. Also provided are methods for reversing inhibition of coagulation.

Description

Attorney Docket No.: 171351.00102 IONIZABLE PILLARARENE ANTICOAGULANT CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No.63 / 578,736 filed on August 25, 2023, the content of which hereby is incorporated by reference herein in its entirety. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0002] This invention was made with government support under R01 GM132345 awarded by the National Institutes of Health. The government has certain rights in the invention. BACKGROUND

[0003] There is an ongoing and unmet need for new approaches for addressing deleterious blood coagulation and reversing the effects of an anti-coagulant. The present disclosure relates to methods and agents to address this need. SUMMARY

[0004] The present disclosure provides methods of using compounds for inhibiting coagulation. In one aspect, the present disclosure provides a method for inhibiting coagulation in a subject in need thereof, the method comprising administering to the subject a compound of formula (I), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereof, whereinAr at each occurrence is independently aryl or heteroaryl; R at each occurrence is independently –L-W; L is a bond or a linker; QB\171351.00102\91698917.1Attorney Docket No.: 171351.00102 W at each occurrence is -OS(O)2O-M+, -OS(O)2OH, a non-sulfate anionic group, a carboxylate group, a phosphonate group, a phosphate group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted aliphatic group, an O-alkyl group, a polyether group, an azide group, H, a substituted or unsubstituted alkyl group, halogen, an amide group, cyano, a substituted or unsubstituted sulfur-containing aliphatic group, nitro, an amino group, a substituted or unsubstituted nitrogen-containing aliphatic group, a substituted or unsubstituted polyethylene glycol group, a polyether group, an O-aryl group, an ester group, a carbamate group, an imine group; an aldehyde group, -SO3H, - SO3-M+, -OSO2F, -OSO2CF3, -OSO2OR′′′, wherein R′′′ is substituted or unsubstituted aryl group or substituted or unsubstituted alkyl group, M+is a counterion; x is 1, 2, 3, 4, 5, or 6; y at each occurrence is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16; at least one y is 1, 2, 3, 4, 5, 6, 7, or 8; and at least one R is –OS(O)2OH or –OS(O)2O-M+.

[0005] In some embodiments, the compound of formula (I) has a structure in which: R at each occurrence is independently –W, –O(CH2)m–W, or –C6H4–W; W at each occurrence is alkyl, alkoxy, O-aryl, S(O)2OH, S(O)2O-M+, OS(O)2OH, OSO2ORa, OS(O)2O-M+, OSO2F, OSO2CF3, C(O)H, C(O)OH, C(O)OR1, C(O)O-M+, P(O)(OH)2, P(O)(OH)(O)-M+, OP(O)(OH)2, OP(O)(OH)(O)-M+, substituted or unsubstituted aryl, substituted or unsubstituted polyethylene glycol, substituted or unsubstituted heteroaryl, azide, halogen, amide, cyano, -S-alkyl, polyether, poly thioether, nitro, amino, C1-10alkylene- NR2R3, polyamines, OC(O)-NR4R5, CHO, imine; Rais substituted or unsubstituted aryl or substituted or unsubstituted alkyl; R1, R2, R3, R4, and R5are each independently C1-10alkyl; and m is 1, 2, 3, 4, 5, 6, 7, or 8.

[0006] The compounds used in the present method include but are not necessarily limited to sulfated pillararenes.

[0007] In an aspect, the present disclosure provides uses of compositions comprising one or more ionizable pillararene(s), comprising one or more ionizable groups. The ionizable groups may be sulfate groups.

[0008] In another aspect, the present disclosure provides a method for reversing anticoagulation effects in a subject in need thereof, the method comprising: administering toAttorney Docket No.: 171351.00102 the subject the compound of formula (I) as described herein, thereby causing anticoagulation effects in the subject; and subsequently administering a nitrogen-based compound to the subject, thereby reversing the anticoagulation effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Further objects, features and advantages of the present disclosure will become apparent from the following detailed description taken in conjunction with the accompanying figures showing illustrative embodiments of the present disclosure. Non-limiting embodiments of the present invention will be described by way of example with reference to the accompanying figures, which are schematic and are not intended to be drawn to scale. In the figures, each identical or nearly identical component illustrated is typically represented by a single numeral. For purposes of clarity, not every component is labeled in every figure, nor is every component of each embodiment of the invention shown where illustration is not necessary to allow those of ordinary skill in the art to understand the invention.

[0010] Figure 1 shows examples of ionizable pillararenes that can be used in a described method.

[0011] Figure 2 shows a crystal structure of P6AS.

[0012] Figure 3 shows X-ray crystal structures of P6AS and P5ACS. a) Cross-eyed stereoview of one molecule of P6AS in the unit cell. Views of the packing of P6AS in the crystal along the b) z-axis and c) y-axis. d) Cross-eyed stereoview of one molecule of P5ACS in the unit cell.

[0013] Figure 4 shows further examples of ionizable pillararenes that can be used in a described method.

[0014] Figure 5 shows a plot of weight versus day number from a maximum tolerated dose (MTD) study performed for P5AS and PBS control. Female Swiss Webster mice (n = 5 for PBS, container up to day 4. N -4 for 6.5 mM group after) were dosed via tail vein on days 0 and 2 (denoted by *) with different concentrations of container or phosphate buffered saline (PBS). The normalized average weight change per study group is indicated. Error bars represent SEM.

[0015] Figure 6 shows plots of aPTT time versus P5AS concentration.

[0016] Figure 7 shows a plot aPTT times versus concentrations for P5AS and Arixtra.

[0017] Figure 8 provides a graph of aPTT time measured for a series of samples containing P5AS (25 μM) and Spermine (0 – 28.7 μM).

[0018] Figure 9 shows a plot of aPTT times versus ionizable pillararene concentrationAttorney Docket No.: 171351.00102 for currently preferred compounds.

[0019] Figure 10 shows a plot of aPTT times versus ionizable pillararene concentration for compounds that perform less effectively than the compounds of Figure 9.

[0020] Figure 11 shows the clotting time (CT) determined by ROTEM assays for negative control, known anticoagulant FondaparinuxTMas positive control, and P5AS. The top graph shows the combined data generated from two separate experiments (solid and empty dots, respectively). The bottom graph shows data generated from a single experiment. Statistical significant difference was determined using t-test. “*” denotes P ≤ 0.05; “**” denotes P ≤ 0.01; “***” denotes P ≤ 0.001; and “****” denotes P ≤ 0.0001.

[0021] Figure 12 shows the dose response curve of P5AS with respect to clotting time.

[0022] Figure 13 shows the CFT, α-angle, and MCF values determined by ROTEM assays for negative control, known anticoagulant FondaparinuxTMas positive control, and P5AS up to 100 micromolar. Data were generated from two separate experiments (solid and empty dots, respectively). DETAILED DESCRIPTION

[0023] Although claimed subject matter will be described in terms of certain examples, other examples, including examples that do not provide all of the benefits and features set forth herein, are also within the scope of this disclosure. Various structural, logical, and process step changes may be made without departing from the scope of the disclosure.

[0024] Ranges of values are disclosed herein. The ranges set out a lower limit value and an upper limit value. Unless otherwise stated, the ranges include the lower limit value, the upper limit value, and all values between the lower limit value and the upper limit value, including, but not limited to, all values to the magnitude of the smallest value (either the lower limit value or the upper limit value).

[0025] As used herein, unless otherwise stated, the term “group” refers to a chemical entity that is monovalent (i.e., has one terminus that can be covalently bonded to other chemical species), divalent, or polyvalent (i.e., has two or more termini that can be covalently bonded to other chemical species). The term “group” also includes radicals (e.g., monovalent and multivalent, such as, for example, divalent, trivalent, and the like, radicals). Illustrative examples of groups include: .the term “aryl group” refers to C5toAttorney Docket No.: 171351.00102 C30, including all integer numbers of carbons and ranges of numbers of carbons therebetween, aromatic or partially aromatic carbocyclic groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, and C30). An aryl group may also be referred to as an aromatic group. The aryl groups can comprise polyaryl groups such as, for example, fused ring or biaryl groups. The aryl group can be unsubstituted or substituted with one or more substituent(s). Examples of substituents include, but are not limited to, various substituents such as, for example, halogens (-F, -Cl, -Br, and -I), azide group, aliphatic groups (e.g., alkyl groups, alkene groups, alkyne groups, and the like), aryl groups, hydroxyl groups, alkoxy groups, carboxylate groups, carboxylic acid groups, ether groups, ester groups, amide groups, thioether groups, thioester groups, and the like, and combinations thereof. A substituent may be or further comprise a sulfonate group or a sulfate group. Examples of aryl groups include, but are not limited to, phenyl groups, biaryl groups (e.g., biphenyl groups and the like), tri-aryl groups (e.g., terphenyl groups and the like), tetra- aryl groups (e.g., tetraphenyl groups and the like), and fused ring groups (e.g., naphthyl groups, anthracene groups, pyrenyl groups, and the like), which may be unsubstituted or substituted.

[0027] As used herein, unless otherwise indicated, the term “heteroaryl group” refers to a monocyclic, polycyclic, bicyclic, or fused ring groups (e.g., aryl groups) comprising one or more aromatic rings comprising at least one heteroatom (e.g., nitrogen, oxygen, sulfur, and the like) in the aromatic ring(s). The heteroaryl groups may be substituted or unsubstituted. Examples of heteroaryl groups include, but are not limited to, benzofuranyl groups, thienyl groups, furyl groups, pyridyl groups, pyrimidyl groups, oxazolyl groups, quinolyl groups, thiophenyl groups, isoquinolyl groups, indolyl groups, triazinyl groups, triazolyl groups, isothiazolyl groups, isoxazolyl groups, imidazolyl groups, benzothiazolyl groups, pyrazinyl groups, pyrimidinyl groups, thiazolyl groups, and thiadiazolyl groups, and the like. Examples of substituents include, but are not limited to, halogens (-F, -Cl, -Br, and -I), aliphatic groups (e.g., alkyl groups, alkenyl groups, alkynyl groups, and the like), aryl groups, alkoxy groups, amine groups, carboxylate groups, carboxylic acids, ether groups, alcohol groups, alkyne groups (e.g., acetylenyl groups and the like), and the like, and combinations thereof.

[0028] As used herein, unless otherwise indicated, the term “aliphatic” refers to branched or unbranched hydrocarbon groups that, optionally, contain one or more degree(s) of unsaturation. Degrees of unsaturation can arise from, but are not limited to, cyclic aliphatic groups. For example, the aliphatic groups / moieties are a C1 to C40 aliphatic group, including all integer numbers of carbons and ranges of numbers of carbons therebetween (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24,Attorney Docket No.: 171351.00102 C25, C26, C27, C28, C29, C30, C31, C32, C33, C34, C35, C36, C37, C38, C39, and C40). Aliphatic groups include, but are not limited to, alkyl groups, alkene groups, and alkyne groups. The aliphatic group can be unsubstituted or substituted with one or more substituent(s). Examples of substituents include, but are not limited to, various substituents such as, for example, halogens (-F, -Cl, -Br, and -I), azide group, aliphatic groups (e.g., alkyl groups, alkene groups, alkyne groups, and the like), aryl groups, hydroxyl groups, alkoxy groups, carboxylate groups, carboxylic acid groups, ether groups, ester groups, amide groups, thioether groups, thioester groups, and the like, and combinations thereof.

[0029] As used herein, unless otherwise indicated, the term “alkyl group” refers to branched or unbranched saturated hydrocarbon groups. Examples of alkyl groups include, but are not limited to, methyl groups, ethyl groups, n- and isopropyl groups, n-, iso-, sec-, and tert- butyl groups, and the like. For example, the alkyl group can be a C1to C12, including all integer numbers of carbons and ranges of numbers of carbons therebetween (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, and C12). The alkyl group can be unsubstituted or substituted with one or more substituent(s). Examples of substituents include, but are not limited to, various substituents such as, for example, halogens (-F, -Cl, -Br, and -I), azide group, aliphatic groups (e.g., alkyl groups, alkene groups, alkyne groups, and the like), aryl groups, hydroxyl groups, alkoxy groups (-OR, where R is an alkyl group), carboxylate groups, carboxylic acid groups, ether groups, ester groups, amide groups, thioether groups, thioester groups, and the like, and combinations thereof.

[0030] As used herein, unless otherwise indicated, the term “amino” refers to both unsubstituted and substituted amines. For example, the amino can be -NRXRY, or -N+RXRYRZ. RX, RY, RZare each independently H, or a substituent as defined herein. Therefore, the amino group may be mono-substituted, disubstituted or trisubstituted; or the amino group may be in the form of a quaternary ammonium cation. The substituents may include, for example, alkyl, alkoxy, silyl, carbonyl, sulfonyl, cycloalkyl, heterocycle, and aryl as defined herein.

[0031] As used herein, the term “subject” can include mammals and non-mammals. Mammals can include, but are not limited to, humans, non-human primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, and swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice, and guinea pigs; and the like. Examples of non-mammals include, but are not limited to, birds, fish and the like. In particular embodiments, the subject is a human or individual.

[0032] The present disclosure provides methods that include administration ofAttorney Docket No.: 171351.00102 ionizable pillararenes (e.g., sulfated pillararenes) to a subject in need of anticoagulation therapy. An ionizable pillararene (e.g., sulfated pillararenes) comprises any compound described in PCT publication no. WO 2021 / 174209, from which the description of ionizable pillararenes (e.g., sulfated pillararenes) is incorporated herein by reference. However, PCT publication no. WO 2021 / 174209 describes use of sulfated pillararenes and related compounds for reversing the effects of agents that are used to treat cardiovascular related disorders, such as coronary thrombosis and embolism. The present disclosure demonstrates that the described pillararenes and related compounds unexpectedly themselves have anti-coagulation properties. To date, the anticoagulant property demonstrated by the present disclosure (such as that of sulfated pillararenes) has not been reported or suggested by other biocompatible container molecules or sequestrants. As such, in some embodiments, the present disclosure includes use of ionizable pillararenes (e.g., sulfated pillararenes) and related compounds as a first administered compound to inhibit blood coagulation, i.e., the described compounds are not administered to the subject to reverse the effect of any other agent.

[0033] The present method can include administration of a macrocyclic arene to a subject in need of anticoagulation therapy. As an example, the macrocyclic arenes described in “Recent advances in the synthesis and applications of macrocyclic arenes” Han et al., Chem. Soc. Rev., 2023, 52, 3265, the entire disclosure of which is incorporated herein by reference, may be modified such that the macrocyclic arenes comprise one or more sulfate groups (e.g., - OSO3H and -OSO3M, where M is a counterion as described herein), such as those described herein.

[0034] The present method can include administration of an extended pillar[n]arene or a Geminiarene to a subject in need of anticoagulation therapy. As an example, the compounds as described in Wu et al., Pillararene-Inspired Macrocycles: From Extended Pillar[n]arenes to Geminiarenes, Accounts of Chemical Research 202255 (21), 3191-3204, the entire disclosure of which is incorporated herein by reference, may be modified such that macrocyclic arenes comprise one or more sulfate groups (e.g., -OSO3H and -OSO3M, where M is a counterion as described herein).

[0035] The present method can also include administration of a terphen[3]arene macrocycle to a subject in need of anticoagulation therapy. The compounds as described in Zhao et al., Water-soluble terphen[3]arene macrocycle: a versatile reversal agent of neuromuscular blockers, Chem. Commun., 2023, 59, 5858, the entire disclosure of which is incorporated herein by reference, may be suitable for this method.

[0036] The present method can also include administration of a water-solubleAttorney Docket No.: 171351.00102 prismarene to a subject in need of anticoagulation therapy. Suitable examples of such compounds are described in Del Regno et al., Molecular Recognition in an Aqueous Medium Using Water-Soluble Prismarene Hosts” Org. Lett.2022, 24, 2711–2715, the entire disclosure of which is incorporated herein by reference. Additionally, it is expected that modifications to these compounds such that the compounds comprise one or more sulfate groups (e.g., -OSO3H and -OSO3M, where M is a counterion as described herein) are suitable for use in a method of the present disclosure.

[0037] The present method can also include administration of a water-soluble prismarene Prism[5]arene to a subject in need of anticoagulation therapy. Suitable examples of such compounds are described in Zhai and Isaacs, Chemistry-A European Journal 2022, 28, e202201743, doi.org / 10.1002 / chem.202201743, the entire disclosure of which is incorporated herein by reference.

[0038] The present disclosure provides ionizable pillararenes (e.g., sulfated pillararenes) for use in anticoagulation therapy. An ionizable pillararene (e.g., sulfated pillararene) comprises a macrocyclic core comprising a plurality of aryl groups, where adjacent aryl groups are covalently connected (e.g., linked) via linking groups, such as, for example, aliphatic linking groups (e.g., -CH2- groups). The linking groups may be oriented ortho (e.g., 1,2-aryl linkages), para (e.g., 1,4-aryl linkages), or meta (e.g., 1,3-aryl linkages) on the aryl groups. In an illustrative example, the linkages may be on different phenylene rings of an aryl group and correspond to an ortho, meta, or para linkage if the different phenylene rings were superimposed. In an illustrative example, when the aryl group is a biphenylene or similar aryl group and linkages such as the following:meta linkage. That is, ortho refers to the orientation of one linking group relative to the other. Non-limiting examples of ionizable pillararenes (e.g., sulfated pillararenes) are provided herein. Non-limiting examples of methods of making ionizable pillararenes (e.g., sulfated pillararenes) are provided herein.

[0039] In one aspect, the present disclosure provides a method for inhibiting coagulation in a subject in need thereof, the method comprising administering to the subject a compound of formula (I), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereof,Attorney Docket No.: 171351.00102 wherein Ar at eachR at each occurrence is independently –L-W; L is a bond or a linker; W at each occurrence is -OS(O)2O-M+, -OS(O)2OH, a non-sulfate anionic group, a carboxylate group, a phosphonate group, a phosphate group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted aliphatic group, an O-alkyl group, a polyether group, an azide group, H, a substituted or unsubstituted alkyl group, halogen, an amide group, cyano, a substituted or unsubstituted sulfur-containing aliphatic group, nitro, an amino group, a substituted or unsubstituted nitrogen-containing aliphatic group, a substituted or unsubstituted polyethylene glycol group, a polyether group, an O-aryl group, an ester group, a carbamate group, an imine group; an aldehyde group, -SO3H, - SO3-M+, -OSO2F, -OSO2CF3, -OSO2OR′′′, wherein R′′′ is substituted or unsubstituted aryl group or substituted or unsubstituted alkyl group, M+is a counterion; x is 1, 2, 3, 4, 5, or 6; y at each occurrence is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14,15, or 16; at least one y is 1, 2, 3, 4, 5, 6, 7, or 8; and at least one R is –OS(O)2OH or –OS(O)2O-M+.

[0040] In some embodiments, L is a bond. In some embodiments, L is –O(CH2)m– or –C6H4–.

[0041] In some embodiments, R at each occurrence is independently –W, –O(CH2)m–W, or –C6H4–W; W at each occurrence is alkyl, alkoxy, O-aryl, S(O)2OH, S(O)2O-M+, OS(O)2OH, OSO2ORa, OS(O)2O-M+, OSO2F, OSO2CF3, C(O)H, C(O)OH, C(O)OR1, C(O)O-M+,Attorney Docket No.: 171351.00102 P(O)(OH)2, P(O)(OH)(O)-M+, OP(O)(OH)2, OP(O)(OH)(O)-M+, substituted or unsubstituted aryl, substituted or unsubstituted polyethylene glycol, substituted or unsubstituted heteroaryl, azide, halogen, amide, cyano, -S-alkyl, polyether, poly thioether, nitro, amino, C1-10alkylene- NR2R3, polyamines, OC(O)-NR4R5, CHO, imine; Rais substituted or unsubstituted aryl or substituted or unsubstituted alkyl; R1, R2, R3, R4, and R5are each independently C1-10alkyl; and m is 1, 2, 3, 4, 5, 6, 7, or 8.

[0042] In some embodiments, M+is Na+, K+, Ca2+, Mg2+, Zn2+, H4N+, Et3NH+, Me4N+, (HOCH2CH2)3NH+, or a cationic form of ethylenediamine, piperazine, or trishydroxymethyl aminomethane (TRIS). In some embodiments, M+is Na+, K+, and H4N+. In particular embodiments, M+is Na+.

[0043] In some embodiments, at least one R is an ionizable group. Examples of ionizable groups include, but are not limited to: -OS(O)2O-M+, -OS(O)2OH, non-sulfate anionic groups, such as sulfonate and corresponding acid (e.g., -O(CH2)mS(O)2O-M+or - O(CH2)mS(O)2OH, where m is 1, 2, 3, 4, 5, 6, 7, or 8; -C6H4S(O)2OH (and the like, where the terminal O is removed (e.g., -CH2S(O)2O-M+or -CH2S(O)2OH)), or -S(O)2O-M+or -S(O)2OH); carboxylate and corresponding acid (e.g., CO2H, -C(O)O-M+, -O(CH2)mC(O)O-M+, or - O(CH2)mC(O)OH, where m is 1, 2, 3, 4, 5, 6, 7, or 8; for example, -OCH2CO2-M+or - OCH2CO2H (and the like, where the terminal O is removed (e.g., -CH2CO2-M+or - CH2CO2H))); phosphonate and corresponding acid groups (e.g., -O(CH2)mP(O)(OH)(O)-M+or -O(CH2)mP(O)(OH)2, where m is 1, 2, 3, 4, 5, 6, 7, or 8; for example, -O(CH2)2P(O)(OH)2(and the like, where the terminal O is removed (e.g., -(CH2)2P(O)(OH)2 or -(CH2)2P(O)(OH)(O)- M)); and phosphate groups (e.g., -OP(O)(OH)2, and the like).

[0044] Additional examples of R groups include, but are not limited to, substituted or unsubstituted aryl groups (e.g., phenyl); substituted or unsubstituted heteroaryl groups (e.g., pyridinyl or pyrazyl); substituted or unsubstituted aliphatic groups (e.g., C1-10alkyl, C1-10alkenyl, C1-10alkynyl, or C3-6cycloalkyl); O-alkyl groups (such as OC1-4alkyl); azide groups (e.g. N3); substituted or unsubstituted alkyl groups (e.g., C1-4haloalkyl and C1-4hydroxyalkyl); halogens (e.g., -Br, -F, -I, -Cl); amide groups (e.g., C(O)NH2); cyano groups (e.g., CN); substituted or unsubstituted sulfur-containing aliphatic groups (e.g., -S-alkyl and poly thioethers, and the like); nitro groups; amino groups (e.g., -NH2or -NHC1-3alkyl); substituted or unsubstituted nitrogen-containing aliphatic groups (e.g., polyamines, aliphatic groups comprising secondary and / or tertiary amines, and the like); substituted or unsubstituted polyethylene glycol groups (e.g., -(OCH2)nOH wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10);Attorney Docket No.: 171351.00102 polyether groups (e.g., -(R6O)nH or -O(R6O)nH wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 and R6is substituted or unsubstituted C1-10alkyl, substituted or unsubstituted C3-12aryl, or substituted or unsubstituted C3-12heteroaryl); O-aryl groups (e.g., aryloxy groups); ester groups (e.g., -C(O)OR6); carbamate groups (e.g., -OC(O)NR7R8wherein R7, R8are each independently C1-10alkyl); imine groups (for example, -C(R9)=NR10or -N=C(R11)(R12) wherein R9, R10, R11, R12are each independently C1-10alkyl); aldehyde groups; -SO3H groups; -SO3Na groups; -OSO2F groups; -OSO2CF3 groups; -OSO2OR′′′ groups (where R′′′ are substituted or unsubstituted aryl groups or substituted or unsubstituted alkyl groups),

[0045] In various embodiments, an ionizable pillararene of formula (I) is provided, in which each Ar is an aryl group and each methylene group is a linking group that links the Ar groups. The orientation of the linking groups on each Ar group may be ortho, meta, or para.

[0046] An ionizable pillararene (e.g., sulfated pillararene) used in a described method can comprise various aryl groups. The aryl groups may all be the same or at least two of the aryl groups are different. Non-limiting examples of aryl groups are independently at each occurrence chosen from phenylene groups, fused-ring groups (e.g., naphthyl groups, anthracenyl groups, phenanthrenyl groups, tetracenyl groups, pentacenyl groups, and the like), biaryl groups (e.g., biphenylene groups and the like), terphenylene groups, tetraphenyl groups, and the like, and combinations thereof. As described herein, a single Ar group of formula (I) can also be a combination of different aryl groups, such as a combination of phenyl and naphthalene. For avoidance of doubt, the aryl groups are bivalent, such as shown in formula (I).

[0047] Adjacent Ar groups of formula (I) (e.g., various aryls as defined herein) can be linked by various linking groups attached at various locations on the Ar groups. In various examples, the linkages on the Ar groups are in a para orientation. In various examples, the linkages on the Ar groups are in a meta orientation. In various examples, the linkages on the Ar groups are in an ortho orientation. As an illustrative example, non-limiting examples of para oriented linkages include: ,Attorney Docket No.: 171351.00102 , linkages are within the scope of

[0048] In some embodiments, Ar at each occurrence is independently phenyl, fused- ring aryl, biaryl, terphenyl, or tetraphenyl. In some embodiments, each Ar is phenyl.

[0049] In some embodiments, the compound of formula (I) is a compound of formula (I-a), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereof (R)y, wherein each y is as defined in formula (I). Invarious examples, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or 32 of the R groups are independently -OS(O)2O-M+or - OS(O)2OH.

[0050] In some embodiments, the compound of formula (I) or formula (I-a) is a compound of formula (I-a’), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereof R , where x is 1, 2, 3, 4, 5, or7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or 32 of the R groups are independently -OS(O)2O-M+or -OS(O)2OH.Attorney Docket No.: 171351.00102

[0051] In various embodiments, the compound of formula (I), formula (I-a), or formula (I-a’) is a compound of formula (I-a-i), (I-a-ii), (I-a-iii), (I-a-iv), or (I-a-v), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereof ,

[0052] In various examples, the compound of formula (I) is an ionizable pillararene (e.g., sulfated pillararene). In particular embodiments, the compound is of formula (I-a-i), (I- a-ii), (I-a-iii), (I-a-iv), or (I-a-v), in which each R is -OS(O)2O-M+or -OS(O)2OH.

[0053] In various embodiments, at least one y is 1, 2, 3, 4, 5, 6, 7, or 8 and at least one R group is -OS(O)2O-M+(e.g., M+is Na+or K+) or -OS(O)2OH. In some embodiments, at least one R is –OS(O)2O-M+.

[0054] In some embodiments, at least two Ar groups each have at least one R, which is independently –OS(O)2OH or –OS(O)2O-M+.

[0055] In some embodiments, each Ar has at least one R that is independently – OS(O)2OH or –OS(O)2O-M+.

[0056] In some embodiments, at least one Ar does not have R that is –OS(O)2OH or – OS(O)2O-M+.

[0057] In some embodiments, the compound is of formula (I-a), wherein each Ar has at least one R that is independently –OS(O)2OH or –OS(O)2O-M+.Attorney Docket No.: 171351.00102

[0058] In some embodiments, the compound is of formula (I-a), wherein at least one Ar does not have R that is –OS(O)2OH or –OS(O)2O-M+.

[0059] An aryl group of the present compounds may comprise one or more phenyl group(s). In various non-limiting examples, at least two, at least three, or at least 4, or all of the one or more phenyl group(s) of one or more of the aryl group(s) comprise at least 1 or at least 2 R groups independently selected from -OS(O)2O-M+and -OS(O)2OH. All of the aryl groups, one or more or all of which may be phenylene group(s), may comprise a sulfate group - OS(O)2O-M+or -OS(O)2OH. In various embodiments, at least one aryl group, which may be a phenylene group, does not comprise a sulfate group (e.g., -OS(O)2O-M+or -OS(O)2OH). In various embodiments, other R groups may be used and are provided herein.

[0060] The R group(s) may be at any position(s) on an Ar group. In the case of an Ar group with multiple R groups, the individual R groups may be at any combination of positions of the Ar group. In various embodiments, all of the Ar groups comprise an R group that is independently -OS(O)2O-M+or -OS(O)2OH. In various embodiments, at least one Ar group does not comprise an R group that is -OS(O)2O-M+or -OS(O)2OH. In various embodiments, the Ar groups may be further substituted with various substituents, such as, for example, -H, alkyl groups, aliphatic groups, polyethylene glycol groups, or the like, or a combination thereof.

[0061] In some embodiments, the compound is: ,Attorney Docket No.: 171351.00102 or, wherein R is SO3Na.can be administered to the subject in need thereof at an effective dose. The effective dose can be measured, for example, by a resultant blood concentration of the administered compound. In some embodiments, the effective dose results in a blood concentration of 500 μM or less in the subject. For example, the effective dose of the administered compound can result in a blood concentration of 450 μM or less, 400 μM or less, 350 μM or less, 300 μM or less, 250 μM or less, 200 μM or less, 150 μM or less, 140 μM or less, 130 μM or less, 120 μM or less, 110 μM or less, or 100 μM or less. In some embodiments, the effective dose of the administered compound results in a blood concentration of 150 μM or less in the subject (such as human or other mammals). For example, the effectiveAttorney Docket No.: 171351.00102 dose of the administered compound can result in a blood concentration of about 20 ^M, about 40 ^M, about 60 ^M, about 80 ^M, about 100 ^M, about 120 ^M, or about 140 μM in the subject.

[0064] In another aspect, the present disclosure provides compositions comprising one or more ionizable pillararene(s) as described herein (e.g., sulfated pillararene(s)) for use in anti- coagulation therapy. Non-limiting examples of compositions are described herein.

[0065] The carrier can be an aqueous carrier suitable for administration to subjects including humans. The carrier can be sterile. The carrier can be a physiological buffer. Examples of suitable carriers include sucrose, dextrose, saline, and / or a pH buffering element (such as, a buffering element that buffers to, for example, a pH from pH 5 to 9, from pH 6 to 8, (e.g., 6.5)) such as histidine, citrate, or phosphate. Additionally, pharmaceutically acceptable carriers may be determined in part by the particular composition being administered. Accordingly, there are a wide variety of suitable formulations of pharmaceutical compositions of the present disclosure. Additional, non-limiting examples of carriers include solutions, suspensions, and emulsions that are dissolved or suspended in a solvent before use, and the like. The composition may comprise one or more diluents. Examples of diluents, include, but are not limited to distilled water, physiological saline, vegetable oil, alcohol, dimethyl sulfoxide, and the like, and combinations thereof. Compositions may contain stabilizers, solubilizers, suspending agents, emulsifiers, soothing agents, buffers, preservatives, and the like, and combinations thereof. Compositions may be sterilized or prepared by sterile procedure. A composition of the disclosure may also be formulated into a sterile solid preparation, for example, by freeze-drying, and may be used after sterilization or dissolution in sterile injectable water or other sterile diluent(s) immediately before use. Additional examples of pharmaceutically acceptable carriers include, but are not limited to, sugars, such as, for example, lactose, glucose, and sucrose; starches, such as, for example, corn starch and potato starch; cellulose, including sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as, for example, peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as, for example, propylene glycol; polyols, such as, for example glycerin, sorbitol, mannitol, and polyethylene glycol; esters, such as, for example, ethyl oleate and ethyl laurate; agar; buffering agents, such as, for example, magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxicAttorney Docket No.: 171351.00102 compatible substances employed in pharmaceutical formulations. Additional non-limiting examples of pharmaceutically acceptable carriers can be found in: Remington: The Science and Practice of Pharmacy (2012) 22ndEdition, Philadelphia, PA. Lippincott Williams & Wilkins. For example, a composition comprises a modified peptide, and a sterile, suitable carrier for administration to subjects including humans—such as a physiological buffer such as sucrose, dextrose, saline, pH buffering (such as from pH 5 to 9, from pH 7 to 8, from pH 7.2 to 7.6, (e.g., 7.4)) element such as, for example, histidine, citrate, or phosphate. In various examples, the composition may be suitable for injection. Parenteral administration includes infusions and injections, such as, for example, intramuscular, intravenous, intraarterial, intraperitoneal, subcutaneous administration, and the like.

[0066] The compositions may be administered systemically. Compositions may be administered orally, may be administered parenterally, and / or intravenously. Compositions suitable for parenteral, administration may include aqueous and / or non-aqueous carriers and diluents, such as, for example, sterile injection solutions. Sterile injection solutions may contain anti-oxidants, buffers, bacteriostatic agents and solutes, which render the composition isotonic with the blood of the intended recipient. Aqueous and / or non-aqueous sterile suspensions may include suspending agents and thickening agents.

[0067] Compositions may be prepared at a patient’s bedside or by a pharmaceutical manufacturer. In the latter case, the compositions can be provided in any suitable container, such as, for example, a sealed sterile vial, ampoule, or the like, and may be further packaged (the combination of which may be referred to as a kit) to include instruction documents for use by a pharmacist, physician, other health care provider, or the like. The compositions can be provided as a liquid, or as a lyophilized or powder form that can be reconstituted if necessary when ready for use. In particular, the compositions can be provided in combination with any suitable delivery form or vehicle, examples of which include but are not limited to liquids, caplets, capsules, tablets, inhalants or aerosol, etc. The delivery devices may comprise components that facilitate release of the pharmaceutical agents over certain time periods and / or intervals, and can include compositions that enhance delivery of the pharmaceuticals, such as nanoparticle, microsphere or liposome formulations, a variety of which are known in the art and are commercially available.

[0068] An ionizable pillararene may comprise one or more counterions (e.g., cations). Examples of cations include, but are not limited to, ions comprising one or more positively charged nitrogen atom(s) (e.g., ammonium ions, primary ammonium ions, secondary ammonium ions, tertiary ammonium ions, quaternary ammonium ions, or a combinationAttorney Docket No.: 171351.00102 thereof, where the non-hydrogen group(s) on the ammonium are chosen from aliphatic groups, alkyl groups, aryl groups, and combinations thereof).

[0069] Compositions of the present disclosure can be administered to any human or non-human animal in need of therapy for coagulation.

[0070] The methods of the disclosure are suitable for use with any subject who has been diagnosed with or is suspected of having a disease or a condition wherein blood clotting is involved and wherein the subject is in need or would benefit from receipt of an anticoagulant. In embodiments, the subject is undergoing or is scheduled to undergo treatment for a blood or organ condition. In embodiments, the subject is undergoing or is a candidate for a surgical procedure. In embodiments, the subject has been diagnosed with or is suspected of having any of disseminated intravascular coagulation, venous thrombosis, deep-vein thrombosis, pulmonary embolism, peripheral arterial embolism, a stroke, acute coronary syndrome, atrial fibrillation, a heart attack, or unstable angina. In embodiments the subject is undergoing prolonged bed rest or hemofiltration, or has an indwelling catheter.

[0071] The present disclosure also provides for inhibiting or stopping the anticoagulation effect of a described compound. In another aspect, the present disclosure provides a method for reversing anticoagulation effects in a subject in need thereof, the method comprising: administering to the subject the compound of formula (I) as described herein, thereby causing anticoagulation effects in the subject; and subsequently administering a nitrogen-based compound to the subject, thereby reversing the anticoagulation effects.

[0072] In various embodiments, when the pillararene is administered, it does not comprise any additional compounds or drugs aside from the pillararene and pharmaceutically acceptable carrier or excipient. In an embodiment, inhibiting, stopping, or reversing the anticoagulation effect comprises administering to the subject an effective amount of one or more nitrogen-based compound comprising one or more positively charged nitrogen atoms. For example, the nitrogen-based compound may comprise one or more hydrophobic groups, such as, for example, one or more aliphatic groups, aryl groups, and / or heteroaryl groups. The nitrogen-based compound may be a monoammonium, a diammonium, a polyammonium, a monopyridinium, a dipyridinium, a polypyridinium, a monoimidazolium, a diimidazolium, a polyimidazolium, a monoguanidinium, a diguanidinium, or a polyguanidinium. Examples of nitrogen-based compound include, but are not limited to, spermine, cadaverine, putrescine, 1,6- hexanediammonium, hexamethonium, decane-1,10-diammonium, decamethonium, trimethyl hexylammonium, trimethyl octylammonium, trimethyl decylammonium, N,N-dimethyl octyl ammonium, hexane-1,6-dipyridinium, octane-1,8-dipyridinium, decane-1,10-dipyridinium,Attorney Docket No.: 171351.00102 protamine, cystamine hydrochloride,

[0074] In some embodiments, the compound of formula (I) is: orAttorney Docket No.: 171351.00102 , wherein R is SO3Na. compound of formula (I) is:, wherein R is SO3Na.known to those skilled in the art can be used to introduce the compounds and / or compositions of the present disclosure to a subject. These methods include, but are not limited to, intravenous, intramuscular, intracranial, intrathecal, intradermal, subcutaneous, oral routes, and the like, and combinations thereof. The dose of the composition comprising a compound and a pharmaceutical agent will necessarily be dependent upon the needs of the subject to whom the composition is to be administered. These factors include, but are not necessarily limited to, the weight, age, sex, medical history, and nature and stage of the disease for which a therapeutic effect is desired, and wherein inhibiting coagulation is desired.

[0077] The steps of the method described in the various examples disclosed herein are sufficient to carry out the methods of the present disclosure. Thus, in an example, the method consists essentially of a combination of the steps of the methods disclosed herein. In another example, the method consists of such steps. EXAMPLES

[0078] The following examples are presented to illustrate the present disclosure. They are not intended to be limiting in any manner.

[0079] Example 1

[0080] This example provides a description of exemplary compounds that can be usedAttorney Docket No.: 171351.00102 in methods of the disclosure.

[0081] General experimental details. Starting materials were purchased from commercial suppliers and were used without further purification or were prepared by literature procedures. Melting points were measured on a Meltemp apparatus in open capillary tubes and are uncorrected. IR spectra were recorded on a JASCO FT / IR 4100 spectrometer and are reported in cm-1.1H NMR spectra were measured on Bruker instruments operating at 400 or 600 MHz for1H and 100 MHz for13C. Mass spectrometry was performed using a JEOL AccuTOF electrospray instrument (ESI). ITC data was collected on a Malvern Microcal PEAQ-ITC instrument.

[0082] Synthetic procedures and characterization data.

[0083] Host P5AS.from methods known in the art. The procedure for the last step was: to a mixture of compound 3 (0.200 g, 0.328 mmol) and pyridine sulfur trioxide complex (1.050 g, 6.56 mmol) was added dry pyridine (10 mL). The resulting mixture was stirred at 90 ºC under N2for 24 hours. The reaction mixture was cooled to RT. The product precipitated out of the solution and was collected by filtration. The solid was slurried in water (5 mL), and the pH was adjusted to 8.4Attorney Docket No.: 171351.00102 by slow addition of saturated aqueous NaHCO3. After addition of EtOH (35 mL), the crude product was collected by centrifugation 7000 rpm × 7 min. The precipitate was suspended in ethanol (20 mL × 2), sonicated for 30 minutes, and solid collected by centrifugation. The crude solid was redissolved in minimum amount of water (2 ml) and purified by size exclusion chromatography using Sephadex® G25 resin (30mm x 200mm) and eluted by water. Pure product was collected as the front fractions. After drying under high vacuum, the compound P5AS was obtained as a white solid (0.374 g, 0.229 mmol, 70 % yield). M.p. > 310oC (decomposed). IR (ATR, cm-1): 3490w, 1630m, 1497m, 1399m, 1234s, 1116s, 1042s, 995m, 941m, 858m, 806m.1H NMR (600 MHz, D2O): 7.31 (s, 10H), 4.00 (s, 10H).13C NMR (150 MHz, D2O, EtOH as internal reference): 147.4, 134.1, 125.6, 30.8. MS (ESI): m / z 791.78179 ([M - 2Na]2-), calculated 791.79597.

[0085] Host P5ACS.2 mL) was treated dropwise with a solution of propane sultone (0.687 g, 5.63 mmol) in acetone (4 mL). This solution was stirred at RT for 5 days (d) and then EtOH (25 mL) was added to the mixture to yield the crude product as a precipitate. The precipitate was obtained by filtration, the solid was dissolved in H2O (0.5 mL), and then re-precipitated by the addition of EtOH (5 mL) to yield P5ACS as a light yellow solid (45 mg, 0.022 mmol, 8 %). M.p. > 300 ºC (decomposed). IR (ATR, cm-1): 3452w, 2936w, 1725m, 1625m, 1479m, 1471m, 1406m, 1181s, 1035s, 951w, 798m, 756m.1H NMR (400 MHz, D2O): 6.76 (s, 10H), 3.90 (m, 10H), 3.86 (s, 10H), 3.68 (m,Attorney Docket No.: 171351.00102 10H), 3.05 (m, 20H), 2.08 (m, 20H).13C NMR (150 MHz, D2O, EtOH as internal reference): δ 150.9, 129.8, 116.7, 68.5, 49.1, 31.1, 25.5. HR-MS (ESI): m / z 1002.03215 ([M - 2Na]2-), calculated 1002.03072.

[0087] Host P6AS.those known in the art. The procedure for the last step was: to a mixture of compound 8 (0.200 g, 0.27 mmol) and pyridine sulfur trioxide complex (1.090 g, 6.83 mmol) was added dry pyridine (10 mL). The resulting mixture was stirred at 70 ºC under N2for 24 hours. The reaction mixture was cooled to RT. The product precipitated out of the solution and was collected by filtration. The solid was slurried in water (5 mL), and the pH was adjusted to 8.4 by slow addition of saturated aqueous NaHCO3. After addition of EtOH (35 mL), the crude product was collected by centrifugation 7000 rpm × 7 min. The precipitate was suspended in ethanol (20 mL × 2), sonicated for 30 minutes, and solid collected by centrifugation. The crude solid was redissolved in minimum amount of water (2 ml) and purified by size exclusion chromatography using Sephadex® G25 resin (30mm x 200mm) and eluted by water. Pure product was collected as the front fractions. After drying under high vacuum, the compound P6AS was obtained as a white solid (0.352 g, 0.18 mmol, 66 % yield). M.p. > 290oC (decomposed). IR (ATR, cm-1): 3509w, 1712m, 1630m, 1498m, 1364m, 1237m, 1113s, 1045s,Attorney Docket No.: 171351.00102 995m, 942m, 861m, 813m.1H NMR (600 MHz, D2O): 7.35 (s, 12H), 4.11 (s, 12H).13C NMR (150 MHz, D2O and CD3OD 10:1): δ 148.1, 133.6, 125.6, 31.5. MS (ESI): m / z 954.7593 ([M- 2Na]2-), calculated 954.7537.

[0089] Host P6S8. OEt OEtwere those known in the art. The procedure for the last step was: to a mixture of compound octahydroxy pillar[6]arene (0.100 g, 0.15 mmol) and pyridine sulfur trioxide complex (0.479 g, 3 mmol) was added dry pyridine (5 mL). The resulting mixture was stirred at 70 ºC under N2 for 24 hours. The reaction mixture was cooled to RT. The product precipitated out of the solution and was collected by filtration. The solid was slurried in water (4 mL), and the pH was adjusted to 8.4 by slow addition of saturated aqueous Na2CO3. After addition of EtOH (10 mL), the crude product was collected by centrifugation 7000 rpm × 7 min. The precipitate was suspended in ethanol (10 mL × 2), sonicated for 30 minutes, and solid collected by centrifugation. The crude solid was redissolved in minimum amount of water (0.5 mL) and purified by size exclusion chromatography using Sephadex® G25 resin (30mm x 200mm) and eluted by water. Pure product was collected as the front fractions. After drying under high vacuum, the compound P6S8 (10) was obtained as a white solid (0.075 g, 0.051 mmol, 33 % yield). M.p. > 285oC (decomposed). IR (ATR, cm-1): 3491w, 1630m, 1440s, 1234s, 1078m, 1043s, 941m, 878m, 800m, 667m.1H NMR (600 MHz, D2O): 7.39 (s, 4H), 7.25 (s, 4H), 7.04 (s, 8H), 4.07 (s, 4H), 3.99 (s, 8H).13C NMR (150 MHz, D2O, EtOH as internal reference): δ 147.7, 147.6, 138.7, 134.2, 133.2, 129.2, 125.3, 125.1, 35.7, 31.1. MS (ESI): m / z 718.88334Attorney Docket No.: 171351.00102 ([M-2Na]2-), calculated 718.88632.

[0091] Host P7AS.those known in the art. The procedure for the last step was: to a mixture of compound (HO)14 pillar[7]arene (0.020 g, 0.023 mmol) and pyridine sulfur trioxide complex (0.375 g, 2.34 mmol) was added dry pyridine (3 mL). The resulting mixture was stirred at 70 ºC under N2 for 24 hours. The reaction mixture was cooled to RT. The product precipitated out of the solution and was collected by filtration. The solid was slurried in water (1 mL), and the pH was adjusted to 8.4 by slow addition of saturated aqueous Na2CO3. After addition of EtOH (10 mL), the crude product was collected by centrifugation 7000 rpm × 7 min. The precipitate was suspended in ethanol (10 mL × 2), sonicated for 30 minutes, and solid collected by centrifugation. The crude solid was redissolved in minimum amount of water (0.5 mL) and purified by size exclusion chromatography using Sephadex® G25 resin (30mm x 200mm) and eluted by water. Pure product was collected as the front fractions. After drying under high vacuum, the compound P7AS was obtained as a white solid (0.025 g, 0.011 mmol, 46 % yield). M.p. > 290oC (decomposed). IR (ATR, cm-1): 3494w, 1624m, 1444s, 1244m, 1102s, 1049s, 995m, 875m, 807m, 614s.1H NMR (600 MHz, D2O): 7.29 (s, 14H), 4.14 (s, 14H).13C NMR (150 MHz, D2O, Dioxane as external reference): δ 147.6, 132.8, 124.7, 30.8. MS (ESI): m / z 547.36023Attorney Docket No.: 171351.00102 ([M-4Na]4-), calculated 547.36080.

[0093] RimP5AS. OBn OBnbased on methods known in the art. The penta-hydroxy pillar[5]arene compound was synthesized by using the methods known in the art. To a mixture of penta-hydroxy pillar[5]arene (0.200 g, 0.328 mmol) and pyridine sulfur trioxide complex (1.050 g, 6.56 mmol) was added dry pyridine (10 mL). The resulting mixture was stirred at 70 ºC under N2for 24 hours. The reaction mixture was cooled to RT. The product precipitated out of the solution and was collected by filtration. The solid was slurried in water (5 mL), and the pH was adjusted to 9 by slow addition of saturated aqueous NaHCO3. Addition of EtOH (EtOH / H2O v / v = 2:1) gave a precipitate which was removed by centrifugation (7000 rpm × 10 min). The filtrate was collected as the crude product and was redissolved in a minimum amount of water (2 mL) and purified by size exclusion chromatography using Sephadex® G25 resin (5 cm × 50 cm) with water as eluent. The front fractions eluting from the column contain pure product. After drying under high vacuum, RimP5AS was obtained as a white solid (0.374 g, 0.229 mmol, 67 % yield, content: ~92%, determined by using sodium 2-bromoethanesulfonate as1H NMR internal standard). M.p. > 300 ˚C (decomposed).1H NMR (400 MHz, D2O): 7.21 (s, 5H), 6.56 (s, 5H), 3.90 (s, 10H), 3.24 (s, 15H).13C NMR (150 MHz, D2O, EtOH as internal reference): 155.3, 143.2, 134.5, 129.6, 124.7, 114.9, 56.7, 30.6.Attorney Docket No.: 171351.00102

[0095] Solubility determination.

[0096] Determination of the solubility of P5AS in water. Compound P5AS was added in excess to 0.5 mL deuterium oxide. This suspension was magnetically stirred at room temperature overnight and then centrifuged (4500 rpm) twice for 10 min each time. Supernatant (50 μL) and sodium 3-(trimethylsilyl)propionate-2,2,3,3-d4 (TMSP) (10 mM, 50 μL in D2O) were added into 0.4 mL deuterium solvent. The concentration of P5AS was measured with1H NMR and calculated using sodium 3-(trimethylsilyl)propionate-2,2,3,3-d4 (TMSP) as internal reference.

[0097] Determination of the solubility of P6AS in water. Compound P6AS was added in excess to 0.5 mL deuterium oxide. This suspension was magnetically stirred at room temperature overnight and then centrifuged (4500 rpm) twice for 10 min each time. Supernatant (50 μL) and sodium 3-(trimethylsilyl)propionate-2,2,3,3-d4(TMSP) (10 mM, 50 μL in D2O) were added into 0.4 mL deuterium solvent. The concentration of P6AS was measured with1H NMR and calculated using sodium 3-(trimethylsilyl)propionate-2,2,3,3-d4(TMSP) as internal reference.

[0098] Example 2

[0099] This Example refers to use of a compound described in Figures 1 and 4.

[0100] A maximum tolerated dose study for P5AS in female Swiss Webster mice were performed at three concentrations (6.5 mM, data shown in Figure 5; 13 mM and 26 mM data not shown). The 13 mM and 26 mM groups experiments were terminated due to unexpected significant side effects and death. Figure 5 provides a graphical summary of data using a 6.5 mM concentration at a volume of 0.150 ml administered to mice with an average weight of 31.5 g. Results using P5AS were similar to P6AS but behavior score was more severe post- injection. Mice showed dose-dependent response to compound. Mice were withdrawn, freezing up, exhibiting ataxia w / physical manipulation, and some labored breathing. Mice returned to baseline in ~1-2h post-injection. Severe latent toxicity likely given that 3 out of 5 of the 26 mM mice died and 2 out of 5 of the 13 mM mice died within 1-3 days post injection. Necropsy analysis indicated P5AS has toxic and anticoagulative properties at higher doses. The 6.5 mM group tolerated second injection significantly better. For the higher dosage groups, mice had accumulation of blood on the tail and the fur. Their carcass was extremely pale and the liver and internal organs were pale. The intestinal tract was filled with dark black ingesta suggestive of digested blood. Both mice had either a coagulopathy or a condition such as hemorrhagic shock. The final concentration of P5AS in the bloodstream of the 6.5 mM group is estimated to be about 123 μM. Thus, in an embodiment, an effective dose as used in a method of thisAttorney Docket No.: 171351.00102 disclosure does not result in a blood concentration of that exceeds about 150 μM.

[0101] Highly water-soluble molecular container compounds (e.g., cyclodextrins, calixarenes, and cucurbiturils) typically display good to very good in vivo biocompatibility (e.g., as determined by maximum tolerated dose studies) and are often used for biomedical applications such as drug delivery and drug sequestration. For example, highly charged molecular containers, such as CaptisolTM( ^-cyclodextrin with sulfonate groups that is used to formulate and deliver insoluble drugs) and Sugammadex (BridionTM^-cyclodextrin with 8 carboxylate group used to sequester neuromuscular blocking agents post surgically) have been used in humans and are known to be biocompatible. Surprisingly, it was demonstrated in the present studies that P5AS has remarkable anticoagulant property, as a high dose of P5AS resulted in death of tested mice likely caused by coagulopathy (a condition in which the blood's ability to coagulate is impaired). To date, anticoagulant property of sulfated pillararene molecules (e.g., P5AS) has not been reported or suggested by other biocompatible container molecules or sequestrants. Therefore, the anticoagulant activity demonstrated by the present compounds (e.g., P5AS) is an unexpected result.

[0102] Example 3

[0103] To confirm the anticoagulative effects of P5AS, Prothrombin Time (PT) and activated partial thromboplastin time (aPTT) assays were performed using pooled human plasma. The results are summarized in the Tables and in Figure 6.

[0104] P5AS did not show any changes in prothrombin time (PT) up to a concentration of 0.1 mM and was therefore deemed to not have any significant effect on the extrinsic pathway. In contrast, P5AS was shown to have a potent effect on the aPTT time (intrinsic pathway) at concentrations greater than 10 μM as shown in Figure 6 relative to blank as control. This experiment was followed up with a more focused concentration range of 1.56 μM to 100 μM in duplicate also shown in Figure 6. The prolongation of aPTT time is comparable to that seen with unfractionated heparin at the 0.06 to 0.5 IU / mL levels (data not shown).

[0105] The effect of P5AS on aPTT time was next compared with the effects with the known anticoagulant Arixtra (fondaparinux). The data is shown in Figure 7. As can be seen from Figure 7, P5AS compares favorably with Arixtra (Fondaparinux) on a concentration basis.

[0106] Example 4

[0107] Given that P5AS (-10 charge) has a potent effect on coagulation, and because it is often important to reverse the effects of anticoagulant compounds, the influence of the addition of the spermine (+4 charge) on aPTT time was determined. It was hypothesized thatAttorney Docket No.: 171351.00102 spermine would bind very tightly to P5AS to form P5AS•spermine complex (now -6 charge) which because of the change in charge should be a less potent anticoagulant. Figure 8 shows the results of aPTT measurements for [P5AS] = 25 μM along with different concentrations of spermine (0 – 28.7 μM). Clearly, at an ≈ 1:1 molar ratio of P5AS:spermine, the aPTT time decrease dramatically back toward the baseline aPTT time for pooled plasma (29.3 seconds). Spermine itself has an effect on aPTT at 287 μM (100 μg / ml) = 75.7 (Table 2). This data establishes conclusively the concept that ammonium compounds like spermine can be used as reversal agents for aPTT prolongation induced by P5AS.

[0108] Example 5

[0109] A series of structural analogues (Figure 4) were analyzed via the PT and aPTT assays. All of the compounds showed only minor changes in the PT assay at concentrations up to 100 μM. Figures 9 and 10 show the aPTT time for the four compounds that had the largest effect on the aPTT time (P5AS, P5ACS, P6AS, P7AS; Figure 9) and separately for the compounds that has smaller effects on the aPTT time (RimP5AS, WP5, P6S8, WP6, DimerACS, TrimerACS4 (Figure 10). These results inform structure activity relationships as follows. Comparison of P5AS with WP5 and P5ACS suggests that sulfate substituents are more advantageous than carboxylate or sulfonate substituents and that the charge density at the rim of the pillararene cavity may be important. Comparison of the aPTT results for P5AS with RimP5AS indicate importance for both of the portals of the pillararene to have charged sulfate substituents. The aPTT results for P5AS, P6AS, and P7AS show that size of the pillar[n]arene skeleton has some influence on aPTT. Comparison of P6AS with P6S8 shows that the number of sulfate substituents and most likely the charge density on each face of the pillararene play an important role in the anti-coagulative properties of P5AS – P7AS. Linear oligomers DimerACS and TrimerACS4 show only small changes in aPTT time up to 100 μM which suggests that the conformational constraints imposed by macrocyclization influence the anti- coagulative properties of P5AS – P7AS.

[0110] The following Tables relate to data presented in this specification.Attorney Docket No.: 171351.00102 Table 1. Data from comparing P5AS and fondaparinux sodium Conc µM P5ASaPTT (sec) conc Arixtra µg / ml Conc arixtra mM aPTT (sec)c)Table 2. Data showing effects of Spermine (NPP17) on 25 μM P5AS NPP17 NPP+25 µM P5AS

[0111] As can be seen from Table 2, when 28.7 μM spermine is used to reverse the effect of 25 μM P5AS there is a marked decrease in aPTT time back toward the “background” aPTT levels.Attorney Docket No.: 171351.00102 Table 3. Data related to Figures 9 and 10 aPTT(sec)Conc µM Blanc P5AS RimP5AS WP5 P5ACS P6AS P6S8 WP6 P7AS 100 28.8 >160 33.2 43.2 83.7 >160 34.6 54.0 109.4 25 29.0 61.2 30.2 35.2 44.9 119.5 30.2 36.7 38.1 6.25 28.8 36.5 29.5 30.9 31.5 48.1 28.8 31.1 30.2 029.0PT(sec)Conc µM Blanc P5AS RimP5AS WP5 P5ACS P6AS P6S8 WP6 P7AS 100 10.8 11.1 11.1 10.5 14.6 11.4 10.4 10.7 11.0 25 10.9 10.6 11.0 10.8 10.9 10.3 10.7 10.9 10.6 6.25 10.9 10.9 10.9 10.7 10.1 10.2 10.8 10.8 10.9 010.9Diluted in NPP2017aPTT(sec)Conc µM Blanc P5AS P5ACS P6AS P7AS100.00 30.5 >160 91.2 >160 9650.00 101.3 62.6 >160 53.325.00 63.4 46.7 126.0 38.812.50 45.4 36.9 71.0 32.46.25 37.3 31.9 47.8 30.53.13 33.1 30.1 37.1 30.01.56 31.4 29.8 32.8 29.60.78 31.0 29.2 31.3 29.80.00 30.4Diluted in NPP2017PT(sec)Conc µM Blanc P5AS P5ACS P6AS P7AS100.00 11.2 11.7 15.0 12.1 10.750.00 11.0 12.4 11.4 10.425.00 10.8 11.4 10.8 10.612.50 10.8 10.6 10.3 10.86.25 11.1 10.3 10.3 11.03.13 11.0 10.3 10.6 11.01.56 11.0 10.4 10.8 11.00.78 11.1 10.5 10.8 11.10.00 11.2Diluted in NPP2017

[0112] Example 6

[0113] Additional studies on the anticoagulant property of P5AS were conducted using rotational thromboelastography (ROTEM) and the results are shown in Figures 11-13. ROTEM provides a viscoelastic assay of hemostasis which allows global measurement of blood clotAttorney Docket No.: 171351.00102 formation and dissolution in real time in whole human blood. ROTEM delivers more information compared to PT and aPTT assays, including clotting time (CT in seconds; time for clot to reach 2 mm amplitude), clot formation time (CFT in seconds; time for clot to reach 20 mm amplitude), maximum clot firmness (MCF in millimeters; peak ), and alpha-angle (in degrees; angle between the central horizontal line and the tangent to the curve through the 20 mm amplitude point).

[0114] Figures 11-12 show the clotting times determined by ROTEM assays for negative control, known anticoagulant FondaparinuxTMas positive control, and P5AS at different concentrations. Fondaparinux and P5AS both show an increase in the clotting time in a dose dependent manner up to 100 micromolar.

[0115] Figure 13 shows the CFT, α-angle (alpha angle), and MCF values determined by ROTEM assays for negative control, known anticoagulant FondaparinuxTMas positive control, and P5AS up to 100 micromolar. The CFT plot shows only small differences in clot formation time between the different concentrations of fondaparinux and P5AS, although P5AS (100 μM) appears to trend toward longer CFT times. The MCF plot shows that maximum clot firmness for fondaparinux (50 and 100 μM) is slightly smaller than that of the control. P5AS (25, 50, 100 μM) also shows a decrease in maximum clot firmness in a dose dependent manner. The MCF value at 100 μM P5AS is smaller than that of fondaparinux 100 μM. The alpha angle measurements show that the formation of the clot is significantly slower for fondaparinux (50, 100 μM) and P5AS (25, 50, 100 μM) relative to the control.

[0116] Figures 11-13 report on the intrinsic coagulation pathway (INTEM test), with CaCl2 and ellagic acid as the activator.

[0117] Although the present disclosure has been described with respect to one or more particular example(s), it will be understood that other examples of the present disclosure may be made without departing from the scope of the present disclosure.

[0118] For reasons of completeness, various aspects of the invention are set out in the following numbered clauses:

[0119] Clause 1. A method for inhibiting coagulation in a subject in need thereof, the method comprising administering to the subject a compound of formula (I), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereof,Attorney Docket No.: 171351.00102 wherein Ar at eachR at each occurrence is independently –L-W; L is a bond or a linker; W at each occurrence is -OS(O)2O-M+, -OS(O)2OH, a non-sulfate anionic group, a carboxylate group, a phosphonate group, a phosphate group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted aliphatic group, an O-alkyl group, a polyether group, an azide group, H, a substituted or unsubstituted alkyl group, halogen, an amide group, cyano, a substituted or unsubstituted sulfur-containing aliphatic group, nitro, an amino group, a substituted or unsubstituted nitrogen-containing aliphatic group, a substituted or unsubstituted polyethylene glycol group, a polyether group, an O-aryl group, an ester group, a carbamate group, an imine group; an aldehyde group, -SO3H, -SO3-M+, -OSO2F, -OSO2CF3, -OSO2OR′′′, wherein R′′′ is substituted or unsubstituted aryl group or substituted or unsubstituted alkyl group, M+is a counterion; x is 1, 2, 3, 4, 5, or 6; y at each occurrence is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16; at least one y is 1, 2, 3, 4, 5, 6, 7, or 8; and at least one R is –OS(O)2OH or –OS(O)2O-M+.

[0120] Clause 2. The method of clause 1, wherein R at each occurrence is independently –W, –O(CH2)m–W, or –C6H4–W; W at each occurrence is alkyl, alkoxy, O-aryl, S(O)2OH, S(O)2O-M+, OS(O)2OH, OSO2ORa, OS(O)2O-M+, OSO2F, OSO2CF3, C(O)H, C(O)OH, C(O)OR1, C(O)O-M+, P(O)(OH)2, P(O)(OH)(O)-M+, OP(O)(OH)2, OP(O)(OH)(O)-M+, substituted or unsubstituted aryl, substituted or unsubstituted polyethylene glycol, substituted or unsubstituted heteroaryl,Attorney Docket No.: 171351.00102 azide, halogen, amide, cyano, -S-alkyl, polyether, poly thioether, nitro, amino, C1-10alkylene- NR2R3, polyamines, OC(O)-NR4R5, CHO, imine; Rais substituted or unsubstituted aryl or substituted or unsubstituted alkyl; R1, R2, R3, R4, and R5are each independently C1-10alkyl; and m is 1, 2, 3, 4, 5, 6, 7, or 8.

[0121] Clause 3. The method of any one of clauses 1-2, wherein Ar at each occurrence is independently phenyl, fused-ring aryl, biaryl, terphenyl, or tetraphenyl.

[0122] Clause 4. The method of any one of clauses 1-3, wherein each Ar is phenyl.

[0123] Clause 5. The method of any one of clauses 1-4, wherein the compound is a compound of formula (I-a), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereof a) whereineach y is independently 1, 2, 3, or 4.

[0124] Clause 6. The method of any one of clauses 1-5, wherein the compound is a compound of formula (I-a-i), (I-a-ii), (I-a-iii), (I-a-iv), or (I-a-v), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereof ii)Attorney Docket No.: 171351.00102 v).

[0125] Clauseat least one R is – OS(O)2O-M+.

[0126] Clause 8. The method of any one of clauses 1-7, wherein at least two Ar groups each have at least one R, which is independently –OS(O)2OH or –OS(O)2O-M+.

[0127] Clause 9. The method of any one of clauses 1-8, wherein each Ar has at least one R that is independently –OS(O)2OH or –OS(O)2O-M+.

[0128] Clause 10. The method of any one of clauses 1-8, wherein at least one Ar does not have R that is –OS(O)2OH or –OS(O)2O-M+.

[0129] Clause 11. The method of clause 5, wherein each Ar has at least one R that is independently –OS(O)2OH or –OS(O)2O-M+.

[0130] Clause 12. The method of clause 5, wherein at least one Ar does not have RAttorney Docket No.: 171351.00102 that is –OS(O)2OH or –OS(O)2O-M+.

[0131] Clause 13. The method of any one of clauses 1-12, wherein M+is Na+, K+, Ca2+, Mg2+, Zn2+, H4N+, Et3NH+, Me4N+, (HOCH2CH2)3NH+, or a cationic form of ethylenediamine, piperazine, or trishydroxymethyl aminomethane (TRIS).

[0132] Clause 14. The method of clause 13, wherein M+is Na+, K+, H4N+, Et3NH+, Me4N+, or (HOCH2CH2)3NH+.

[0133] Clause 15. The method of clause 14, wherein M+is Na+.

[0134] Clause 16. The method of any one of clauses 1-15, wherein x is 3, 4, 5, or 6.

[0135] Clause 17. The method of clause 1, wherein the compound is: orisAttorney Docket No.: 171351.00102 , wherein R is SO3Na. for reversing anticoagulation effects in a subject in needthereof, the method comprising: administering to the subject the compound of formula (I) according to any one of clauses 1-18, thereby causing anticoagulation effects in the subject; and subsequently administering a nitrogen-based compound to the subject, thereby reversing the anticoagulation effects.

[0138] Clause 20. The method of clause 19, wherein the nitrogen-based compound is a monoammonium, a diammonium, a polyammonium, a monopyridinium, a dipyridinium, a polypyridinium, a monoimidazolium, a diimidazolium, a polyimidazolium, a monoguanidinium, a diguanidinium, or a polyguanidinium.

[0139] Clause 21.The method of any one of clauses 19-20, wherein the nitrogen-based compound is spermine, cadaverine, putrescine, 1,6-hexanediammonium, hexamethonium, decane-1,10-diammonium, decamethonium, trimethyl hexylammonium, trimethyl octylammonium, trimethyl decylammonium, N,N-dimethyl octyl ammonium, hexane-1,6- dipyridinium, octane-1,8-dipyridinium, decane-1,10-dipyridinium, protamine, cystamine ,isspermine.

Claims

Attorney Docket No.: 171351.00102 CLAIMS What is claimed is:

1. A method for inhibiting coagulation in a subject in need thereof, the method comprising administering to the subject a compound of formula (I), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereof, whereinAr at each occurrence or R at each occurrence is independently –L-W; L is a bond or a linker; W at each occurrence is -OS(O)2O-M+, -OS(O)2OH, a non-sulfate anionic group, a carboxylate group, a phosphonate group, a phosphate group, a substituted or unsubstituted aryl group, a substituted or unsubstituted heteroaryl group, a substituted or unsubstituted aliphatic group, an O-alkyl group, a polyether group, an azide group, H, a substituted or unsubstituted alkyl group, halogen, an amide group, cyano, a substituted or unsubstituted sulfur-containing aliphatic group, nitro, an amino group, a substituted or unsubstituted nitrogen-containing aliphatic group, a substituted or unsubstituted polyethylene glycol group, a polyether group, an O-aryl group, an ester group, a carbamate group, an imine group; an aldehyde group, -SO3H, -SO3-M+, -OSO2F, -OSO2CF3, -OSO2OR′′′, wherein R′′′ is substituted or unsubstituted aryl group or substituted or unsubstituted alkyl group, M+is a counterion; x is 1, 2, 3, 4, 5, or 6; y at each occurrence is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16; at least one y is 1, 2, 3, 4, 5, 6, 7, or 8; and at least one R is –OS(O)2OH or –OS(O)2O-M+.Attorney Docket No.: 171351.00102 2. The method of claim 1, wherein R at each occurrence is independently –W, –O(CH2)m–W, or –C6H4–W; W at each occurrence is alkyl, alkoxy, O-aryl, S(O)2OH, S(O)2O-M+, OS(O)2OH, OSO2ORa, OS(O)2O-M+, OSO2F, OSO2CF3, C(O)H, C(O)OH, C(O)OR1, C(O)O-M+, P(O)(OH)2, P(O)(OH)(O)-M+, OP(O)(OH)2, OP(O)(OH)(O)-M+, substituted or unsubstituted aryl, substituted or unsubstituted polyethylene glycol, substituted or unsubstituted heteroaryl, azide, halogen, amide, cyano, -S-alkyl, polyether, poly thioether, nitro, amino, C1-10alkylene- NR2R3, polyamines, OC(O)-NR4R5, CHO, imine; Rais substituted or unsubstituted aryl or substituted or unsubstituted alkyl; R1, R2, R3, R4, and R5are each independently C1-10alkyl; and m is 1, 2, 3, 4, 5, 6, 7, or 8.

3. The method of claim 1, wherein Ar at each occurrence is independently phenyl, fused-ring aryl, biaryl, terphenyl, or tetraphenyl.

4. The method of claim 1, wherein each Ar is phenyl.

5. The method of claim 1, wherein the compound is a compound of formula (I-a), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereof a) whereineach y is independently 1, 2, 3, or 4.

6. The method of claim 1, wherein the compound is a compound of formula (I-a-i), (I-a- ii), (I-a-iii), (I-a-iv), or (I-a-v), or a salt, a partial salt, a hydrate, a polymorph, a stereoisomer, conformational isomer, or a mixture thereofAttorney Docket No.: 171351.00102 v).Attorney Docket No.: 171351.00102 7. The method of claim 1, wherein at least one R is –OS(O)2O-M+.

8. The method of claim 1, wherein at least two Ar groups each have at least one R, which is independently –OS(O)2OH or –OS(O)2O-M+.

9. The method of claim 1, wherein each Ar has at least one R that is independently – OS(O)2OH or –OS(O)2O-M+.

10. The method of claim 1, wherein at least one Ar does not have R that is –OS(O)2OH or –OS(O)2O-M+.

11. The method of claim 5, wherein each Ar has at least one R that is independently – OS(O)2OH or –OS(O)2O-M+.

12. The method of claim 5, wherein at least one Ar does not have R that is –OS(O)2OH or –OS(O)2O-M+.

13. The method of claim 1, wherein M+is Na+, K+, Ca2+, Mg2+, Zn2+, H4N+, Et3NH+, Me4N+, (HOCH2CH2)3NH+, or a cationic form of ethylenediamine, piperazine, or trishydroxymethyl aminomethane (TRIS).

14. The method of claim 13, wherein M+is Na+, K+, H4N+, Et3NH+, Me4N+, or (HOCH2CH2)3NH+.

15. The method of claim 14, wherein M+is Na+.

16. The method of claim 1, wherein x is 3, 4, 5, or 6.

17. The method of claim 1, wherein the compound is:Attorney Docket No.: 171351.00102 or18. The method of claim 1, wherein the compound isAttorney Docket No.: 171351.00102 19. A method for reversing anticoagulation effects in a subject in need thereof, the method comprising: administering to the subject the compound of formula (I) according to any one of claims 1-18, thereby causing anticoagulation effects in the subject; and subsequently administering a nitrogen-based compound to the subject, thereby reversing the anticoagulation effects.

20. The method of claim 19, wherein the nitrogen-based compound is a monoammonium, a diammonium, a polyammonium, a monopyridinium, a dipyridinium, a polypyridinium, a monoimidazolium, a diimidazolium, a polyimidazolium, a monoguanidinium, a diguanidinium, or a polyguanidinium.

21. The method of claim 19, wherein the nitrogen-based compound is spermine, cadaverine, putrescine, 1,6-hexanediammonium, hexamethonium, decane-1,10-diammonium, decamethonium, trimethyl hexylammonium, trimethyl octylammonium, trimethyl decylammonium, N,N-dimethyl octyl ammonium, hexane-1,6-dipyridinium, octane-1,8- dipyridinium, decane-1,10-dipyridinium, protamine, cystamine hydrochloride, ,22. The method of claim 21, wherein the nitrogen-based compound is spermine.

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