WO2012166705A2 - Nanofibres et hydrogels supramoléculaires à base de conjugués acide aminé-nucléobase-glucoside - Google Patents

Nanofibres et hydrogels supramoléculaires à base de conjugués acide aminé-nucléobase-glucoside Download PDF

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WO2012166705A2
WO2012166705A2 PCT/US2012/039821 US2012039821W WO2012166705A2 WO 2012166705 A2 WO2012166705 A2 WO 2012166705A2 US 2012039821 W US2012039821 W US 2012039821W WO 2012166705 A2 WO2012166705 A2 WO 2012166705A2
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hydrogelator
kpa
hydrogel
supramolecular structure
nanofibers
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WO2012166705A3 (fr
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Bing Xu
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Brandeis University
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Publication of WO2012166705A3 publication Critical patent/WO2012166705A3/fr
Priority to US14/093,974 priority Critical patent/US10093674B2/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H13/00—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids
    • C07H13/02—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids
    • C07H13/04—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids having the esterifying carboxyl radicals attached to acyclic carbon atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A61K47/183—Amino acids, e.g. glycine, EDTA or aspartame
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/70—Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
    • A61L31/12—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L31/125—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A61L31/129—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix containing macromolecular fillers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • Hydrogels which consist of crosslinked matrices and water, have emerged as an important class of biomaterials due to their morphological similarity to extracellular matrices (ECM) in tissues and organs.
  • ECM extracellular matrices
  • both natural polymers e.g., collagen, gelatin, hyaluronic acid, and alginate
  • synthetic polymers e.g., poly(D-L-lactide-co-glycolide), poly(N-isopropyl acrylic amide), and poly(ethylene oxide)
  • biomedical applications e.g., tissue engineering and drug delivery
  • the currently known members of each class have considerable drawbacks or limitations.
  • the separation and purification of natural polymers are non-trivial, and synthetic polymers are largely passive even if they are functionalized.
  • Nanofibers comprised of self-assembled peptides, that form supramolecular hydrogels have shown considerable promise. These self-assembled peptides have served as scaffolds to guide the differentiation of neuron progenitor cells, media for cell culture, and carriers for drug release. Like modified peptides, derivatives of glycosides can also self- assemble into nanofibers to give supramolecular gels or hydrogels, which has led to the development of semi-wet peptide/protein arrays as biosensors and intelligent soft materials. Recently, nanofibers of deoxynucleic acid (DNA) were found to form supramolecular hydrogels.
  • DNA deoxynucleic acid
  • the invention relates to a hydrogelator of Formula I
  • cytosinyl is cytosinyl, guaninyl, adeninyl, thyminyl, uracilyl, or an oligonucleic acid; is fructosyl, galactosyl, glucosyl, mannosyl, or an oligosaccharide;
  • R is H or alkyl
  • R 1 is H, alkyl, alkylthioalkyl, aralkyl, heteroaralkyl, hydroxyaralkyl, H0 2 C-alkyl, or guanidinylalkyl;
  • n 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
  • the invention relates to a supramolecular structure, comprising a plurality of any one of the aforementioned hydrogelators.
  • the invention relates to a hydrogel, wherein the hydrogel comprises a plurality of any one of the aforementioned hydrogelators; and water.
  • the invention relates to a hydrogel, wherein the hydrogel comprises a plurality of any one of the aforementioned supramolecular structures; and water.
  • Figure 1 depicts (a) structures of exemplary hydrogelators (except 1C) comprising nucleobase, amino acid, and glycoside; and (b) a cartoon representing the resulting supramolecular structure.
  • Figure 2 depicts an exemplary synthetic route for the preparation of hydrogelators 1A and 2A.
  • Figure 3 depicts the molecular structures and exemplary synthetic routes for the preparation of hydrogelators 1A, 2 A, 2C, 1G, 2G, and compound 1C.
  • Figure 5 depicts transmission electron micrographs of negative stained hydrogels of IT, 2T, 2C, 1A, 2 A, 1G and 2G; and solution of 1C.
  • Scale bar 100 nm; the concentration and pH value for each of them are same as in Figure 4.
  • Figure 6 depicts transmission electron micrographs of hydrogels of (a) IT and (b)
  • Figure 7 depicts transmission electron micrographs of (a) the solution of 1C and (b) the hydrogel of 2C.
  • Figure 8 depicts transmission electron micrographs of hydrogels of (a) 1A and (b)
  • Figure 9 depicts transmission electron micrographs of hydrogels of (a) 1G and (b)
  • Figure 10 depicts the UV-vis absorption spectrum of: (A) IT in aqueous solution (c
  • Figure 11 depicts (A) the critical strain, and (B) dynamic storage moduli (C) of hydrogels of IT, 2T, 2C, 1A, 2A, 1G and 2G; (C) the CD spectra of hydrogels of IT, 1A, 1G and solution of 1C; (D) the CD spectra of hydrogels of 2T, 2C, 2A, and 2G.
  • concentration and pH value for each of them are same as for Figure 4.
  • Figure 12 depicts the circular dichroism (CD) spectra of the hydrogel of IT, the solution of poly(lOA), the mixture solution of thymine acetic acid with poly(lOA) in 1 : 1 molecular ratio, and the hydrogel of IT mixed with poly(lOA) in 1 : 1 molecular ratio.
  • CD circular dichroism
  • Figure 13 depicts (A) strain dependence of dynamic storage moduli (G') and loss moduli (G") of hydrogels of IT, 2T, 2C, 1A, 2A, 1G and 2G; (B) frequency dependence of dynamic storage moduli (G') and loss moduli (G") of hydrogels of IT, 2T, 2C, 1A, 2A, 1G and 2G, as shown in Figure 4.
  • Figure 14 depicts 72 hr cell viability test of (A) hydrogelators 1, and (B) hydrogelators 2; optical images of the scratch-wound assay to assess the effects of 2T in the media on wound closure; optical images of HeLa cells on the surface at 0 h (C); and at 20 h (D) after the creation of wound in the presence of 2T (by adding 500 ⁇ of 2T in the media); and (E) the time-dependent course of digestions of hydrogelators of 2T, 2C, 2A and 2G by proteinase K.
  • Figure 15 depicts the CD spectra of the hydrogels of IT, 1A, 1G, and the solution of
  • Figure 17 depicts (left) strain dependence of dynamic storage moduli (G') and loss moduli (G") of the hydrogels of IT, and lT+deoxyadenosine (A 10 ) mixed gel; (right) frequency dependence of dynamic storage moduli (G') and loss moduli (G") of the hydrogels of IT, and lT+deoxyadenosine (A 10 ) mixed gel shown in Figure 16.
  • Figure 18 depicts (A) The molecular structures of NapFFCGLDD and thymine-FF, and (B) their time-dependent course of the digestions by proteinase K as control experiment, in which NapFFCGLDD is the heptapeptide derivative and thymine-FF is the nucleopeptide without D-glucosamine in conjugation.
  • Figure 19 depicts fluorescence and bright field microscopy images illustrating nuclear localization of DNA released from the IT and nucleic acid complex.
  • Nucleic acid was labeled with fluorescien dye (FITC) (green). Cell nuclei were stained with SYTO 85.
  • FITC fluorescien dye
  • A 500 ⁇ IT and 0.1 ⁇ nucleic acid labeled with FITC complex incubated with HeLa cells for 24 h.
  • B 0.1 ⁇ nucleic acid labeled with FITC incubated with HeLa cells for 24 h.
  • Figure 20 depicts the molecular structures and exemplary synthetic routes for the preparation of hydrogelators of the invention.
  • Figure 21 depicts the molecular structures of hydrogelators consisting of nucleobase, RGD peptides, and glycoside, (a) 1A + RGD; (b) 1C + RGD; (c) 1G + RGD; (d) IT + RGD.
  • the invention relates to a hydrogelator, wherein the hydrogelator comprises, consists essentially of, or consists of a nucleobase, an amino acid, and a glycoside.
  • the invention relates to a multifunctional, biocompatible supramolecular nanofiber or hydrogel, comprising, consisting essentially of, or consisting of an aforementioned hydrogelator.
  • the invention relates to a hydrogelator, comprising a nucleobase (e.g., thymine), an amino acid (e.g., phenylalanine), and a glycoside (e.g., D- glucosamine), wherein they are covalently tethered.
  • the hydrogelator forms molecular nanofibers that result in a supramolecular hydrogel at pH of about 7.0, and concentration of about 3.0 wt%.
  • the invention relates to a nano fiber, comprising a plurality of said hydrogelators.
  • the invention relates to a supramolecular hydrogel, comprising a plurality of said nanofibers.
  • the invention relates to a hydrogelator, wherein the hydrogelator comprises thymine, cytosine, adenine, or guanine.
  • the invention relates to a hydrogelator, wherein the hydrogelator comprises alanine, isoleucine, leucine, methionine, phenylalanine, tryptophan, tyrosine, or valine.
  • the hydrogelator comprises phenylalanine.
  • the hydrogelator comprises diphenylalanine.
  • the invention relates to a hydrogelator, wherein the hydrogelator comprises a glycoside.
  • the inclusion of glycoside in the hydrogelator significantly enhances its resistance to proteases.
  • the glycoside is or is derived from a glucosamine or a galactosamine.
  • the invention relates to a hydrogelator, wherein the hydrogelator does not inhibit the growth of mammalian cells.
  • the invention relates to a nanofiber or a hydrogel comprising any one of the aforementioned hydrogelators.
  • Figure 1 shows the molecular design of two exemplary types of hydrogelators (1 and
  • Hydrogelator 1 consists of a nucleobase (e.g., thymine, cytosine, adenine, or guanine), a phenylalanine, and a D-glucosamine; hydrogelator 2 consists of a nucleobase, a bis(phenylalanine), and a D-glucosamine.
  • the nucleobase and the D- glucosamine connect to the amino acid(s) at the N-terminus and C-terminus, respectively.
  • Figure 2 outlines an exemplary synthetic route for making these hydrogelators.
  • thymine acetic acid (3) is activated by N-hydroxysuccinimide (NHS) before reacting with L- Phe to afford 4.
  • NHS N-hydroxysuccinimide
  • 4 couples with D-glucosamine to give the hydrogelator IT.
  • the addition of the second phenylalanine to 4 affords 5, which couples with D-glucosamine to yield the hydrogelator 2T.
  • 1C starts from protected nucleobases (e.g., (A ⁇ -fos-Boc-cytosine-l-yFi-acetic acid, f-bis- Boc-adenine-9-yl)-acetic acid, and (N 2 -£zs-Boc-guanine-9-yl)-acetic acid).
  • protected nucleobases e.g., (A ⁇ -fos-Boc-cytosine-l-yFi-acetic acid, f-bis- Boc-adenine-9-yl)-acetic acid, and (N 2 -£zs-Boc-guanine-9-yl)-acetic acid).
  • bis(tert- butyloxycarbonyl) (bis-Boc) protected adenine (A ⁇ -bis-Boc-adenine- -yFi-acetic acid (6), was synthesized. After being activated by NHS, 6 reacts with L-Phe to afford 7, which undergoes the
  • protonation and deprotonation of an amine group may be used to dissolve any one of the aforementioned hydrogelators at low pH.
  • hydrogelation may be triggered by increasing the pH.
  • the hydrogelators dissolve in water at about 3.0 wt% and pH of about 10.0.
  • the hydrogelators dissolve in water at about 3.0 wt% and pH of about 10.0 with gentle heating.
  • changing the pH values of the solutions of IT and 2T from 10.0 to 7.0 and 8.5, respectively results in transparent hydrogels.
  • the mixture containing 1C remains as a solution at the same conditions.
  • each hydrogelator exhibits distinctive morphology of the nanostructures in the corresponding hydrogels.
  • the nanofibers of IT are thin and straight and with the diameter of about 12 nm
  • the nanofibers of 2T (about 15 nm in diameter) appear to bend easily and to crosslink relatively heavily.
  • the solution of 1C only results in featureless aggregates, likely due to non-specific absorption of the 1C on the carbon film of TEM grid.
  • the hydrogel of 2C consists of nanobelts (about 25 nm wide) that physically crosslink into networks.
  • the nanofibers of 2C also form bundles that likely contribute to the high storage modulus (Figure 1 IB). While both short nanofibers (14 nm in width and 200 nm in length) and nanoparticles (average diameter of 18 nm) present as the solid phase in the hydrogel of 1A, the hydrogel of 2A exhibits only nanofibers, which tend to crosslink physically to afford the network.
  • the hydrogel of 1G appears to comprise thin nanofibers (9 nm in width) and aggregated nanoparticles whose diameters are about 27 nm.
  • Hydrogelator 2G self-assembles in water to form long thin nano fibers with a width of about 13 nm, and the nano fibers in 2G entangle with each other to form a dense nanofiber network, which also contributes to its relatively high storage modulus (Figure 1 IB).
  • hydrogels are viscoelastic they resist external destruction. Rheometry was used to study the viscoelastic properties of the instant hydrogels and to evaluate their critical strains and storage moduli (G'). Based on the results from the strain sweep (Figure 10), the hydrogel of IT shows the highest tolerance to external shear force with critical strain value at 0.5 % ( Figure 11 A). The critical strain values of the hydrogels of 1A, 1G, 2T, 2C, 2A, and 2G are at 0.23, 0.28, 0.31, 0.27, 0.39, and 0.18 %, respectively, suggesting that the networks in these hydrogels lose their integrity relatively easily upon application of external force.
  • the frequency sweep shows that the dynamic storage moduli (G') of the hydrogels (IT, 2T, 2C, 1A, 2A, 1G and 2G) dominate their dynamic loss moduli (G") ( Figure 10), indicating that all samples behave as viscoelastic materials.
  • the hydrogel of 2C exhibits the highest storage modulus (220 KPa).
  • the hydrogels of 1G, 2G, IT, 2T, and 1A possess relatively high storage moduli of 139, 101, 34, 32, and 6 KPa, respectively.
  • the hydrogel of 2A exhibits the lowest storage modulus (0.37 KPa), likely due to the short constituent nanofibers and nanoparticles, which disfavour the formation of crosslinked network.
  • Circular dichroism (CD) spectroscopy provides helpful information about self- assembled superstructures in the gel phase or liquid crystal phases.
  • CD circular dichroism
  • hydrogels of IT, 2T, 2C, 2A, 2G all exhibit a positive peak near 195 nm and a negative peak around 210 nm, suggesting that the backbones of the hydrogelators adopt ⁇ -sheet-like configurations in the self-assembled structures.
  • the CD spectrum of the hydrogel of 2T shows a negative broad band around 296 nm, which likely originates from the formation of a mesophase of 2T because it locates far from the chromophoric absorption region (ca. 268 nm) of compound 2T ( Figure 10).
  • the CD spectra of hydrogels of 1A and 1G display a maximum around 201 nm and a minimum near 210 nm, slightly red-shifted from the maxima and minima found in typical ⁇ -sheets, indicating that the supramolecular structures share the common feature of a ⁇ -sheet structure, but in a less ordered conformation or in a mixture of ⁇ -sheet and random coil structures.
  • the solution of 1C exhibits the weakest CD absorptions, agreeing with the poor tendency for compound 1C to self-assemble in water to form ordered structures.
  • hydrogelators 1 and 2 were added into a culture of mammalian cells, and the proliferation of the cells was measured. MTT assay results, shown in Figure 14A and Figure 14B, revealed cell viability remained at 90% after incubation with 500 ⁇ hydrogelator (IT, 1G, 2T, 2C, 2A, 2G) for 72 hours. Although the cell viability decreased slightly when the cells were incubated with 500 ⁇ of 1C or 1A for 72 hours, the value of IC 50 is still > 500 ⁇ . These results suggest that hydrogelators 1 and 2 are biocompatible.
  • we also conducted a simple wound-healing assay with hydrogelator IT As shown in Figure 14D, the presence of the hydrogelator of IT in cell culture has little inhibitory effect on the migration of cells.
  • hydrogelators 2 Besides biocompatibility, biostability is also an essential prerequisite for a biomaterial.
  • proteinase K a powerful protease that hydro lyzes a broad spectrum of peptides.
  • hydrogelators 2 exhibit excellent resistance to enzymatic digestion, indicated by more than 85% of 2T and 2G and 95% of 2C and 2A remaining intact after 24 hours of incubation. Due to their high resistance to proteases, the hydrogels formed by hydrogelators 2 promise to serve as new biomaterials for applications that require long-term biostability.
  • an element means one element or more than one element.
  • a reference to "A and/or B", when used in conjunction with open-ended language such as “comprising” can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.
  • the phrase "at least one,” in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements.
  • This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase "at least one" refers, whether related or unrelated to those elements specifically identified.
  • At least one of A and B can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.
  • compositions of the present invention may exist in particular geometric or stereoisomeric forms.
  • polymers of the present invention may also be optically active.
  • the present invention contemplates all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)- isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention.
  • Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.
  • a particular enantiomer of compound of the present invention may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers.
  • the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomers.
  • the invention relates to a hydrogelator of Formula I
  • cytosinyl is cytosinyl, guaninyl, adeninyl, thyminyl, uracilyl, or an oligonucleic acid; is fructosyl, galactosyl, glucosyl, mannosyl, or an oligosaccharide;
  • R is H or alkyl
  • R 1 is H, alkyl, alkylthioalkyl, aralkyl, heteroaralkyl, hydroxyaralkyl, H0 2 C-alkyl, or guanidinylalkyl;
  • n 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
  • the invention relates to any one of the aforementioned
  • hydrogelators wherein S cytosinyl.
  • the invention relates to
  • the invention relates to any one of the
  • hydrogelators wherein is thyminyl.
  • the hydrogelators wherein is thyminyl.
  • invention relates to any one of the aforementioned hydrogelators, wherein is uracilyl. In certain invention relates to any one of the aforementioned hydrogelato n oligonucleic acid.
  • the invention relates to any one of the aforementioned hydrogelato glucosyl.
  • the invention relates to any one of the aforementioned hydrogelators, wherein Vs —- is an oligosaccharide.
  • the invention relates to any one of the aforementioned hydrogelators, ndrosinyl.
  • the invention relates to any one of the aforementioned hydrogelators, wherein R is H.
  • the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is H.
  • the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is alkyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is methyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is ethyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is propyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is isopropyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is butyl.
  • the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is isobutyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is sec-butyl.
  • the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is alkylthioalkyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is CH 3 -S-CH 2 CH 2 -.
  • the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is aralkyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is benzyl.
  • the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is heteroaralkyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is indolyl-CH 2 -. In certain mbodiments, the invention relates to any one of the aforementioned hydrogelators, wherein
  • the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is hydroxyaralkyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is hydroxybenzyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is 4-hydroxybenzyl.
  • the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is H0 2 C-alkyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is HO 2 C-CH 2 -.
  • the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is guanidinylalkyl. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein R 1 is guanidinyl-CH 2 CH 2 CH 2 -.
  • the invention relates to any one of the aforementioned hydrogelators, wherein n is 1. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein n is 2. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein n is 3. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein n is 4. In certain embodiments, the invention relates to any one of the aforementioned hydrogelators, wherein n is 5.
  • the invention relates to a compound selected from the group consisting of:
  • the invention relates to a supramolecular structure, comprising a plurality of any one of the aforementioned hydrogelators.
  • the invention relates to any one of the aforementioned supramolecular structures, wherein the supramolecular structure is in the form of nanofibers or nanobelts.
  • the average diameter of the nanofibers or the average width of the nanobelts is about 8 nm, about 9 nm, about 10 nm, about 11 nm, about 12 nm, about 13 nm, about 14 nm, about 15 nm, about 16 nm, about 17 nm, about 18 nm, about 19 nm, about 20 nm, about 21 nm, about 22 nm, about 23 nm, about 24 nm, or about 25 nm.
  • the nanofibers are crosslinked.
  • the nanofibers are substantially straight. In certain embodiments, the nanofibers are bent. In certain embodiments, the nanofibers form bundles of nanofibers. In certain embodiments, the nanofibers are about 100 nm, about 120 nm, about 140 nm, about 160 nm, about 180 nm, about 200 nm, about 220 nm, about 240 nm, about 260 nm, about 280 nm, or about 300 nm in length.
  • the invention relates to any one of the aforementioned supramolecular structures, wherein the supramolecular structure is in the form of aggregated nanoparticles.
  • the average diameter of the aggregated nanoparticles is about 20 nm, about 21 nm, about 22 nm, about 23 nm, about 24 nm, about 25 nm, about 26 nm, about 27 nm, about 28 nm, about 29 nm, about 30 nm, about 31 nm, about 32 nm, about 33 nm, about 34 nm, or about 35 nm.
  • the invention relates to a hydrogel, wherein the hydrogel comprises a plurality of any one of the aforementioned hydrogelators; and water.
  • the invention relates to a hydrogel, wherein the hydrogel comprises a plurality of any one of the aforementioned supramolecular structures; and water.
  • the invention relates to any one of the aforementioned hydrogels, wherein the hydrogel is formed from a solution of the hydrogelators in water.
  • the hydrogelator is present in an amount from about 1.5% to about 6% by weight. In certain embodiment, the hydrogelator is present in an amount of about 2.0%, about 2.5%), about 3.0%, about 3.5%, or about 4.0% by weight.
  • the invention relates to any one of the aforementioned hydrogels, wherein the hydrogel is formed from a solution of the hydrogelators in water.
  • the pH of the solution is about 10.0, about 9.5, about 9.0, or about 8.5.
  • the invention relates to any one of the aforementioned hydrogels, wherein the hydrogel is formed from a solution of the hydrogelators in water.
  • the temperature of the solution is about 20 °C, about 25 °C, about 30 °C, about 35 °C, or about 40 °C.
  • the invention relates to any one of the aforementioned hydrogels, wherein the hydrogel is formed by decreasing the pH of the solution of hydrogelators in water.
  • the pH at which the supramolecular structure is formed is about 9.0, about 8.5, about 8.0, about 7.5, about 7.0, about 6.5, about 6.0, about 5.5, about 5.0, about 4.5, about 4.0, or about 3.5.
  • the invention relates to any one of the aforementioned hydrogels, wherein the hydrogel has a critical strain value of from about 0.15% to about 0.45%. In certain embodiments, the invention relates to any one of the aforementioned hydrogels, wherein the hydrogel has a critical strain value of about 0.15%, about 0.16%, about 0.17%, about 0.18%, about 0.19%, about 0.20%, about 0.21%, about 0.22%, about 0.23%, about 0.24%, about 0.25%, about 0.26%, about 0.27%, about 0.28%, about 0.29%, about 0.30%, about 0.31%, about 0.32%, about 0.33%, about 0.34%, about 0.35%, about 0.36%, about 0.37%, about 0.38%, about 0.39%, about 0.40%, about 0.41%, about 0.42%, about 0.43%, about 0.44%, or about 0.45%.
  • the invention relates to any one of the aforementioned hydrogels, wherein the hydrogel has a storage modulus of from about 0.2 KPa to about 150 KPa. In certain embodiments, the invention relates to any one of the aforementioned hydrogels, wherein the hydrogel has a storage modulus of about 0.2 KPa, about 0.3 KPa, about 0.4 KPa, about 0.5 KPa, about 0.6 KPa, about 0.8 KPa, about 1 KPa, about 2 KPa, about 3 KPa, about 4 KPa, about 5 KPa, about 6 KPa, about 7 KPa, about 8 KPa, about 9 KPa, about 10 KPa, about 15 KPa, about 20 KPa, about 25 KPa, about 30 KPa, about 35 KPa, about 40 KPa, about 50 KPa, about 60 KPa, about 70 KPa, about 80 KPa, about 90 KPa, about 100 KPa, about
  • the invention relates to any one of the aforementioned hydrogels, wherein the hydrogel is substantially biocompatible. In certain embodiments, the invention relates to any one of the aforementioned hydrogels, wherein the hydrogel is substantially biostable.
  • the invention relates to a method of growing cells, comprising contacting a plurality of cells with any one of the aforementioned supramolecular structures or any one of the aforementioned hydrogels.
  • the cells are engineered tissue cells.
  • the cells are stem cells.
  • the cells are skin cells.
  • the invention relates to a method of delivering a substance to a cell, comprising
  • the invention relates to any one of the aforementioned methods, wherein the substance is a drug. In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the substance is a protein. In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the substance is a gene. In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the substance is siR A. In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the substance is microRNA. In certain embodiments, the invention relates to any one of the aforementioned methods, wherein the substance is a second cell.
  • the invention relates to a method of binding a nucleic acid, comprising
  • the invention relates to any one of the aforementioned methods, wherein the nucleic acid binding is selective nucleic acid binding.
  • the invention relates a method of separating a protein from a substance, comprising
  • the invention relates to any one of the aforementioned methods, wherein the mixture comprises at least two proteins.
  • the invention relates to a method of treating or preventing a viral infection, comprising
  • the invention relates to a method of treating or preventing cancer, comprising
  • the invention relates to a method of preventing adhesion of an organism or a cell to a surface, comprising
  • FIG. 3 depicts six synthetic schemes for various compounds of the invention.
  • Thymine-Phe (4) Thymine acetic acid (184 mg, 1 mmol) and NHS (126.5 mg, 1.1 mmol) were dissolved in 20 mL of DMF, and DCC (226.6 mg, 1.1 mmol) was added to the above solution with stirring. After the reaction mixture was stirred at room temperature overnight, and the resulted solid was filtered off. The filtrate was evaporated under reduced pressure to dryness, and the crude product was used in the next reaction without purification.
  • Thymine-Phe-glucosamine (IT).
  • Compound 4 (331.3 mg, 1 mmol) and NHS (126.5 mg, 1.1 mmol) were dissolved in DMF (30 mL), and DCC (226.6 mg, 1.1 mmol) was added to the above solution with stirring. After the reaction mixture was stirred at room temperature for 12 h, the resulted solid was filtered off, and the filtrate was evaporated under reduced pressure to dryness. The crude product was used for the next reaction without purification.
  • CD spectra were recorded (185-350 nm) using a JASCO 810 spectrometer under a nitrogen atmosphere.
  • the hydrogels (0.2 mL, 3.0 wt %) were placed evenly on the 1 mm thick quartz curvet and scanned with 0.5 nm interval.
  • Figure 10, Figure 11, Figure 12, and Figure 15 depict various UV-Vis and CD spectra.
  • the critical strain ( ⁇ ) value was determined from the storage-strain profiles of the hydrogel sample.
  • the strain applied to the hydrogel sample increased from 0.1 to 100 % (10 rad/s and 25°C). Over a certain strain, a drop in the elastic modulus was observed, and the strain amplitude at which storage moduli just begins to decrease by 5 % from its maximum value was determined and taken as a measure of the critical strain of the hydrogels, which correspond to the breakdown of the cross-linked network in the hydrogel sample.
  • HeLa cells were re-suspended in 10 cm tissue culture dish after washing cells once with PBS. 0.8 mL 0.25 % trypsin containing 0.1 % EDTA was then added, and the cells were re-suspended with 1.6mL complete medium. 5000 cells (in 100 medium) were plated into each vial on a 96 well plate to create a confluent monolayer. After adherent for 24 hr, a wound was created by scraping the cell monolayer with a p200 pipet tip. The cells were washed once with 100 of complete medium to remove flowing cells and replace with 100 of complete medium. 0 hr image was acquired as a reference point.
  • the medium was replaced with 100 ⁇ ⁇ of medium containing 500 ⁇ of hydrogelator IT and the plate was incubate at 37 °C, 5 % C0 2 for 20 hr. 0 hr and 20 hr images were acquired at the match photographed region.
  • HeLa cells were seeded in 2 well chamber slide at a density of 10,000 cell/well. After allowing the attachment at 37°C for 4 h, we removed culture medium and applied ImL of culture medium containing 0.1 ⁇ fluorescein (FITC) labeled poly(lOA) with or without 500 ⁇ hydrogelator IT. After incubation at 37°C for 24 h, we removed the culture medium, washed the cells by ImL PBS for 3 times, then resin the cells in ImL PBS. Fluorescence images were taken by using confocal fluorescence microscope. Figure 19.
  • FITC fluorescein

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Abstract

L'invention concerne des promoteurs d'hydrogel qui comportent une nucléobase, un acide aminé et un glucoside. Ces promoteurs d'hydrogel présentent une résistance accrue aux protéases et peuvent former par auto-assemblage des nanofibres ou des hydrogels supramoléculaires. Ces structures supramoléculaires peuvent être utilisées pour favoriser la croissance de cellules ou pour délivrer une substance à une cellule.
PCT/US2012/039821 2011-05-31 2012-05-29 Nanofibres et hydrogels supramoléculaires à base de conjugués acide aminé-nucléobase-glucoside WO2012166705A2 (fr)

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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2014138367A1 (fr) * 2013-03-06 2014-09-12 Brandeis University Inhibition de la croissance tumorale avec des agrégats de petites molécules
US10308682B2 (en) 2013-03-06 2019-06-04 Brandeis University Inhibition of tumor growth with aggregates of small molecules
US10232037B2 (en) 2014-02-03 2019-03-19 Brandeis University Supramolecular hydrogel of fMLF-based molecules and use thereof
US10857243B2 (en) 2014-04-09 2020-12-08 Brandeis University Enzymatically responsive magnetic particles and their use
WO2016205459A3 (fr) * 2015-06-16 2017-01-26 The Johns Hopkins University Hydrogels thérapeutiques à base de nanofibres pour le traitement local des maladies liées au cerveau
US11160875B2 (en) 2015-06-16 2021-11-02 The Johns Hopkins University Therapeutic nanofiber hydrogels for local treatment of brain-related diseases
US11839661B2 (en) 2017-08-15 2023-12-12 Brandeis University Rapid formation of supramolecular hydrogels by short peptide and bioactive small molecules
US11191724B2 (en) 2017-09-18 2021-12-07 Brandeis University Branched peptides for enzymatic assembly and mitochondria drug delivery
US11834517B2 (en) 2017-09-18 2023-12-05 Brandeis University Branched peptides for enzymatic assembly and mitochondria drug delivery

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