Peptoid polymers and methods of use

Abstract

The present invention provides peptoid polymers and salts thereof that include hydrophobic and polar peptoid monomers and that can reduce or inhibit ice crystal formation at temperatures below 0°C. Also provided are peptoid-peptide hybrids and salts thereof that include the peptoid polymers provided herein. The peptoid polymers, peptoid-peptide hybrids, and salts thereof provided herein can be used to prepare cryoprotective solutions. The peptoid polymers, peptoid-peptide hybrids, salts thereof, and cryoprotective solutions provided herein can be used to prepare antifreeze solutions, frozen foods, and beauty care products. Also provided herein are methods for preserving tissues, organs, cells, or biomacromolecules using the compositions described herein.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 62 / 484,714, filed on April 12, 2017, the disclosure of which is incorporated herein by reference in its entirety for all purposes. Background Art

[0003] Cryoprotective agents (CPAs) are compounds that, when present in a solution, can reduce or inhibit the formation of ice crystals in a solution exposed to temperatures below 0° C. Current CPAs include small molecules (often referred to as osmotic CPAs), synthetic polymers, and antifreeze proteins.

[0004] Organ transplantation is currently the best treatment for end-stage organ failure based on survival, quality of life, and cost-effectiveness considerations. Unfortunately, there is a huge gap between the supply and demand of organ transplants, and this gap is one of the major medical barriers that forces patients with debilitating diseases to suffer a low quality of life during long waiting times. The apparent lack of organs is due to the large amount of waste caused by the lack of reliable preservation methods. In fact, more than 50% of lungs, pancreases, and hearts are still not collected from deceased donors.

[0005] In order to properly preserve the organ, it must be flushed with a preservation solution to remove blood and stabilize the organ. Even if it has been stabilized in a preservation solution, the time available for organ distribution, transportation, and transplantation after being taken out from the donor is also very limited (about 6-12 hours). This short timeliness causes most organs to be supplied only to local patients, because it is usually impossible to determine the matching of remote patients within the limited available time. Due to this shortage, although almost all countries have introduced laws prohibiting the sale of human organs, illegal organ trading and human trafficking have increased to meet demand.

[0006] In general, the osmotic CPAs currently used for organ preservation include ethylene glycol, 1,2-propylene glycol, dimethyl sulfoxide, formamide, glycerol, sucrose, lactose, and D-mannitol. In order to reduce or inhibit ice crystal growth at organ preservation temperatures, the effective concentration of osmotic CPA must be very high (usually requiring ≥60%). At such high concentrations, these compounds can be toxic to the tissue being preserved, and the removal of large amounts of CPA after warming before transplantation can lead to irreversible cell death.

[0007] Other CPAs for reducing or inhibiting ice crystal formation include synthetic polymers and antifreeze proteins. Similar to permeable CPAs, each has its disadvantages. For example, synthetic polymers cannot penetrate cell membranes. Therefore, synthetic polymer CPAs can only control ice formation outside the cell. In order to effectively preserve biological samples, ice crystal formation both inside and outside the cell must be controlled. Naturally occurring antifreeze proteins, such as those isolated from fish, plants, or insects, are very effective in preventing ice formation, but currently available antifreeze proteins are of low purity and very expensive. In addition, the use of antifreeze proteins to preserve biological samples introduces a potential source of immunogenicity.

[0008] Therefore, there is a need in the art for new non-toxic compounds for effectively reducing or inhibiting ice crystal formation at temperatures below 0° C. and cryogenic temperatures. The present disclosure satisfies this need and also provides other advantages. Summary of the invention

[0009] In one aspect, the present invention provides a peptoid polymer or a salt thereof comprising subunits, wherein the subunits include one or more first hydrophobic peptoid monomers H and one or more first polar peptoid monomers P, which are arranged so that the peptoid polymer has a sequence [H a P b ] n or [P b H a ] n , wherein the subscript a representing the number of consecutive first hydrophobic peptoid monomers in the subunit is between 1 and 10; the subscript b representing the number of consecutive first polar peptoid monomers in the subunit is between 1 and 10; and the subscript n representing the number of subunits in the peptoid polymer is between 2 and 50.

[0010] In some embodiments, the peptoid polymer or its salt further comprises substituents X and Y, such that the peptoid polymer has the sequence X-[H a P b ] n -Y or X-[P b H a ] n -Y, wherein X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 Carboxyalkyl, and halogen, or alternatively, X and Y together form a covalent bond.

[0011] In some embodiments, the subunit further comprises a second hydrophobic peptoid monomer and / or a second polar peptoid monomer, such that the peptoid polymer has the sequence [H a P b H c P d ] n or [P b H a P d H c ] n , wherein the subscript c representing the number of consecutive second hydrophobic peptoid monomers in the subunit is between 0 and 10; the subscript d representing the number of consecutive second polar peptoid monomers in the subunit is between 0 and 10; and c and d are not both zero.

[0012] In some embodiments, the peptoid polymer or its salt further comprises substituents X and Y such that the peptoid polymer has the sequence X-[H a P b H c P d ] n -Y or X-[P b H a P d H c ] n -Y, wherein X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 Carboxyalkyl, and halogen, or alternatively, X and Y together form a covalent bond.

[0013] In some embodiments, the peptoid polymer or its salt further includes a sequence Z, wherein the sequence Z includes one or more hydrophobic peptoid monomers and / or one or more polar peptoid monomers, wherein Z is located before the first subunit, after the last subunit, and / or between one or more subunits. In some instances, Z includes one or more hydrophobic peptoid monomers. In other instances, Z includes one or more polar peptoid monomers. In some other instances, Z includes one or more hydrophobic peptoid monomers and one or more polar peptoid monomers.

[0014] In some embodiments, n is between 2 and 10. In other embodiments, a is between 1 and 5. In some embodiments, b is between 1 and 5. In some examples, a is between 1 and 3 and b is between 1 and 3. In some embodiments, c is between 0 and 5. In other embodiments, d is between 0 and 5.

[0015] In another aspect, the present invention provides a peptoid polymer or a salt thereof, comprising: (a) a subunit comprising two first hydrophobic peptoid monomers H and two first polar peptoid monomers P, and (b) two second hydrophobic peptoid monomers located at the C-terminus of the peptoid polymer, arranged such that the peptoid polymer has a sequence [H2P2] n H2 or [P2H2] n H2, wherein the subscript n representing the number of subunits in the peptoid polymer is between 1 and 50. In some embodiments, n is between 1 and 10. In some embodiments, the peptoid polymer or a salt thereof includes Compound 81.

[0016] In some embodiments, the peptoid polymer or its salt further comprises substituents X and Y such that the peptoid polymer has the sequence X-[H2P2] n H2-Y or X-[P2H2] n H2-Y, wherein X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 Carboxyalkyl, and halogen, or alternatively, X and Y together form a covalent bond.

[0017] In some embodiments, the first and / or second hydrophobic peptoid monomers are independently selected from the group consisting of: wherein the subscript m is the number of repeating units and is between 1 and 10.

[0018] In some embodiments, none of the polar peptoid monomers include an optionally substituted C 1-18 A hydroxyalkyl side chain.

[0019] In some embodiments, the first and / or second polar peptoid monomers each include a side chain independently selected from (C 1-6 Alkoxy) (C 1-6alkylene), (oligo[ethylene glycol]), (4- to 10-membered heterocycloalkyl) (C 1-6 alkylene), and (5- to 10-membered heteroaryl) (C 1-6 In some embodiments, (4- to 10-membered heterocycloalkyl) (C 1-6 (4- to 10-membered heterocycloalkyl) (C 1-6 In some examples, the peptoid polymer includes In a specific embodiment, the polar peptoid monomers are all In some embodiments, the peptoid polymer includes Compound 63, Compound 76, Compound 86, or Compound 87.

[0020] In some embodiments, (5- to 10-membered heteroaryl) (C 1-6 (5- to 10-membered heteroaryl) (C 1-6 In some examples, the peptoid polymer includes In a particular example, the peptoid polymer includes compound 73.

[0021] In some embodiments, the side chain includes a methoxyethyl group. In some examples, the peptoid polymer includes In a particular example, the peptoid polymer includes compound 62.

[0022] In some embodiments, the side chain includes an oligo(ethylene glycol) moiety. In some embodiments, the oligo(ethylene glycol) moiety is a 2-(2-(2-methoxyethoxy)ethoxy)ethyl moiety. In some examples, the peptoid polymer includes In a particular example, the peptoid polymer includes compound 67.

[0023] In some embodiments, the peptoid polymer or its salt comprises a polar peptoid monomer having a side chain comprising a hydroxyl group. In some embodiments, the first and / or second polar peptoid monomer is independently selected from the group consisting of:

[0024]

[0025] wherein the subscript m is the number of repeating units and is between 1 and 10.

[0026] On the other hand, the present invention provides a peptoid polymer or a salt thereof, which comprises one or more hydrophobic peptoid monomers and one or more polar peptoid monomers, wherein the one or more polar peptoid monomers each comprise a hydrophobic peptoid monomer selected independently from (C 1-6 Alkoxy) (C 1-6 alkylene), (oligo[ethylene glycol]), (4- to 10-membered heterocycloalkyl) (C 1-6 alkylene), and (5- to 10-membered heteroaryl) (C 1-6 A side chain in the group consisting of an alkylene group.

[0027] In some embodiments, the one or more hydrophobic peptoid monomers are each independently selected from the group consisting of:

[0028] wherein the subscript m is the number of repeating units and is between 1 and 10.

[0029] In some embodiments, (4- to 10-membered heterocycloalkyl) (C 1-6 (4- to 10-membered heterocycloalkyl) (C 1-6 In some examples, the peptoid polymer includes In a specific embodiment, the polar peptoid monomers are all In some embodiments, the peptoid polymer includes Compound 63, Compound 76, Compound 86, or Compound 87.

[0030] In some embodiments, (5- to 10-membered heteroaryl) (C 1-6 (5- to 10-membered heteroaryl) (C 1-6 In some examples, the peptoid polymer includes In a particular example, the peptoid polymer includes compound 73.

[0031] In some embodiments, the side chain includes a methoxyethyl group. In some cases, the peptoid polymer includes In certain instances, the peptoid polymer comprises compound 62.

[0032] In some embodiments, the side chain includes an oligo(ethylene glycol) moiety. In some embodiments, the oligo(ethylene glycol) moiety is a 2-(2-(2-methoxyethoxy)ethoxy)ethyl moiety. In some examples, the peptoid polymer includes In a particular example, the peptoid polymer includes compound 67.

[0033] In some embodiments, the peptoid polymer or its salt further comprises substituents X and Y located at the N-terminus and C-terminus of the peptoid polymer, respectively, wherein X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 Carboxyalkyl, and halogen, or alternatively, X and Y together form a covalent bond.

[0034] In some embodiments, about 10% of the peptoid monomers are hydrophobic. In some embodiments, about 20% of the peptoid monomers are hydrophobic. In some embodiments, about 30% of the peptoid monomers are hydrophobic. In some embodiments, about 40% of the peptoid monomers are hydrophobic. In some embodiments, about 50% of the peptoid monomers are hydrophobic. In some embodiments, about 60% of the peptoid monomers are hydrophobic. In some embodiments, about 70% of the peptoid monomers are hydrophobic. In some embodiments, about 80% of the peptoid monomers are hydrophobic. In some embodiments, about 90% of the peptoid monomers are hydrophobic.

[0035] In some embodiments, the peptoid polymer forms a helical structure. In other embodiments, the peptoid polymer or its salt reduces or inhibits ice crystal formation at a temperature of about 0°C to about -20°C. In some embodiments, the peptoid polymer or its salt reduces or inhibits ice crystal formation at a temperature of about -20°C to about -40°C. In some embodiments, the peptoid polymer or its salt reduces or inhibits ice crystal formation at a temperature of about -20°C. In some other embodiments, the peptoid polymer or its salt reduces or inhibits ice crystal formation at a temperature of about -40°C to about -200°C. In some embodiments, the salt of the peptoid polymer is selected from the group consisting of hydrochloride, acetate, sulfate, phosphate, maleate, citrate, methanesulfonate, nitrate, tartrate, gluconate, and combinations thereof. In some embodiments, the peptoid polymer is not a peptoid polymer described in Tables 2-9.

[0036] On the other hand, the present invention provides a peptoid-peptide hybrid or a salt thereof comprising a peptoid polymer of the present invention and one or more amino acids, wherein the one or more amino acids are located at one or both ends of the peptoid polymer, and / or between one or more subunits. In some embodiments, the peptoid-peptide hybrid is not a peptoid-peptide hybrid described in SEQ ID NO: 3, 4, 5, or 6. In some embodiments, the one or more amino acids are selected from the group consisting of alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, arginine, lysine, leucine, methionine, asparagine, proline, glutamine, serine, threonine, valine, tryptophan, tyrosine, and combinations thereof. In a specific embodiment, the one or more amino acids are selected from the group consisting of isoleucine, leucine, serine, threonine, alanine, valine, arginine, and combinations thereof. In some embodiments, the peptoid-peptide hybrid salt is selected from the group consisting of hydrochloride, acetate, sulfate, phosphate, maleate, citrate, methanesulfonate, nitrate, tartrate, gluconate, and combinations thereof.

[0037] On the other hand, the present invention provides a cryoprotectant comprising a peptoid polymer of the present invention or a salt thereof, a peptoid-peptide hybrid of the present invention or a salt thereof, or a combination thereof. In some embodiments, the cryoprotectant further comprises a compound selected from the group consisting of: an ionic substance, a permeable cryoprotectant, a non-permeable cryoprotectant, an antioxidant, a cell membrane stabilizing compound, a water channel protein or other channel forming compound, an alcohol, a sugar, a sugar derivative, a non-ionic surfactant, a protein, dimethyl sulfoxide (DMSO), polyethylene glycol (PEG), polypropylene glycol (PPG), Polyvinyl pyrrolidone, polyvinyl alcohol, hyaluronic acid, formamide, natural or synthetic hydrogels, and combinations thereof.

[0038] In some examples, the cryoprotectant further includes an alcohol selected from the group consisting of propylene glycol, ethylene glycol, glycerol, methanol, butylene glycol, adonitol, ethanol, trimethylene alcohol, diethylene glycol, polyethylene oxide, erythritol, sorbitol, xylitol, polypropylene glycol, 2-methyl-2,4-pentanediol (MPD), mannitol, inositol, dithiothitol, 1,2-propylene glycol, and combinations thereof.

[0039] In some instances, the cryoprotectant further comprises a sugar selected from the group consisting of monosaccharides, disaccharides, polysaccharides, and combinations thereof. In some instances, the sugar is a monosaccharide selected from the group consisting of glucose, galactose, arabinose, fructose, xylose, mannose, 3-O-methyl-D-pyranose glucose, and combinations thereof. In other instances, the sugar is a disaccharide selected from the group consisting of sucrose, trehalose, lactose, maltose, and combinations thereof. Still in other instances, the sugar is a polysaccharide selected from the group consisting of raffinose, dextran, and combinations thereof.

[0040] In other examples, the cryoprotectant further comprises PEG or PPG having an average molecular weight of less than about 3,000 g / mol. In certain instances, the PEG or PPG has an average molecular weight between about 200 g / mol and 400 g / mol.

[0041] In some examples, the cryoprotectant further comprises a protein selected from the group consisting of bovine serum albumin, human serum albumin, gelatin, and combinations thereof. In other examples, the cryoprotectant further comprises a natural or synthetic hydrogel containing chitosan, hyaluronic acid, or a combination thereof. In still other examples, the cryoprotectant further comprises a nonionic surfactant selected from the group consisting of polyoxyethylene lauryl ether, polysorbate 80, and combinations thereof.

[0042] In another aspect, the present invention provides methods for preserving tissues, organs, or cells. In some embodiments, the method comprises contacting tissues, organs, or cells with a peptoid polymer of the present invention or a salt thereof, a peptoid-peptide hybrid of the present invention or a salt thereof, a cryoprotectant of the present invention, or a combination thereof. In some embodiments, the tissue is a bioengineered tissue. In other embodiments, the tissue, organ, or cell is selected from the group consisting of: heart, liver, lung, kidney, pancreas, intestine, thymus, cornea, nerve cell, platelet, sperm cell, oocyte, genitourinary cell, embryonic cell, stem cell, human pluripotent stem cell, hematopoietic stem cell, lymphocyte, granulocyte, immune system cell, bone cell, primary cell, organoid, and a combination thereof.

[0043] In some embodiments, the peptoid polymer or its salt, the peptoid-peptide hybrid or its salt, the cryoprotectant, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature within about 0°C to about -20°C. In some embodiments, the peptoid polymer or its salt, the peptoid-peptide hybrid or its salt, the cryoprotectant, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature within about -20°C to about -40°C. In some embodiments, the peptoid polymer or its salt, the peptoid-peptide hybrid or its salt, the cryoprotectant, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at about -20°C. In other embodiments, the peptoid polymer or its salt, the peptoid-peptide hybrid or its salt, the cryoprotectant, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature within about -40°C to about -200°C (e.g., -196°C). In certain embodiments, the concentration of the peptoid polymer or its salt, and / or the peptoid-peptide hybrid or its salt in the cryoprotectant is between about 100 nM and about 1,000 mM. In certain embodiments, the concentration of the peptoid polymer or its salt, and / or the peptoid-peptide hybrid or its salt in the cryoprotectant is between about 1 and 100 mM (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 mM).

[0044] In yet another aspect, the present invention provides methods for preserving biomacromolecules. In some embodiments, the method comprises contacting the biomacromolecule with a peptoid polymer of the present invention or a salt thereof, a peptoid-peptide hybrid of the present invention or a salt thereof, a cryoprotectant of the present invention, or a combination thereof. In some embodiments, the biomacromolecule is selected from the group consisting of nucleic acids, amino acids, proteins, isolated proteins, peptides, lipids, complex structures, and combinations thereof.

[0045] In another aspect, the present invention provides a cosmetic care product comprising the peptoid polymer of the present invention or a salt thereof, the peptoid-peptide hybrid of the present invention or a salt thereof, the cryoprotectant solution of the present invention, or a combination thereof.

[0046] On the other hand, the present invention provides antifreeze products such as deicing or ice suppression products, which include peptoid polymers of the present invention or salts thereof, peptoid-peptide hybrids of the present invention or salts thereof, cryoprotectants of the present invention, or combinations thereof. In some embodiments, antifreeze products are used to prevent, inhibit, or delay the formation of ice on or inside an object, including but not limited to aircraft or parts thereof, gas pipelines, windows, electrical equipment, drones, cables (e.g., power lines), and mechanical equipment (e.g., automobile engines, gear systems, brake systems, etc.), etc.

[0047] In still another aspect, the present invention provides a frozen food comprising a peptoid polymer of the present invention or a salt thereof, a peptoid-peptide hybrid of the present invention or a salt thereof, a cryoprotectant of the present invention, or a combination thereof. In some embodiments, the frozen food is selected from the group consisting of ice cream, yogurt, seafood, fruit, and meat products.

[0048] Other objects, features, and advantages of the present invention will become apparent to those skilled in the art from the following detailed description and accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 A general scheme for the synthesis of peptoid oligomers using the "submonomer" approach is shown.

[0050] Figure 2A and 2B Shown are the results of a capillary cryoassay performed at -20°C. Figure 2A Compounds 1 (1 eq) and 10 (1 eq) were dissolved in MilliQ water and subjected to sub-zero temperature assays. Comparisons were made with solutions of water and ethylene glycol (EG) (18 eq) alone. Figure 2B show Figure 2A Normalized results of the assays shown in .

[0051] Figures 3A-3D X-ray diffraction (XRD) crystallographic data are shown. Figure 3A Shown are XRD data for a solution containing 5 mM compound 12 and 17.5% (v / v) ethylene glycol (EG). Figure 3B XRD data for a solution containing 30% (v / v) EG is shown. Figure 3C XRD data for a solution containing 17.5% (v / v) EG is shown. Figure 3D The ice ring fractions of several solutions containing EG, compound 2 (labeled as "B"; SEQ ID NO: 10), compound 12 (labeled as "D"; SEQ ID NO: 8), and / or compound 8 (labeled as "E"; SEQ ID NO: 9) are shown. For each different solution, two different ice ring fractions were determined.

[0052] Figures 4A-4G Shown are X-ray diffraction (XRD) crystallographic data for solutions containing 5 mg / mL Compound 10, Compound 12, Compound 8, Compound 13, Compound 11, and Compound 58 compared to an ethylene glycol (EG) control. Each solution also contained 300 mM NaCl, 100 mM HEPES, 15% (v / v) ethylene glycol, and the pH was adjusted to 7.2. Figure 4A : XRD crystallography (left) and spectrum (right) of compound 10. Figure 4B: XRD crystallography (left) and spectrum (right) of compound 12. Figure 4C : XRD crystallography (left) and spectrum (right) of compound 8. Figure 4D : XRD crystallography (left) and spectrum (right) of compound 13. Figure 4E : XRD crystallography (left) and spectrum (right) of compound 11. Figure 4F : XRD crystallography (left) and spectrum (right) of compound 58. Figure 4G : EG control XRD crystallography (left) and spectra (right). For the XDR spectra, the intensity is plotted as a function of angle (2θ degrees).

[0053] Figure 5A-5G Shown are X-ray diffraction (XRD) crystallographic data for solutions containing 1 mg / mL Compound 10, Compound 12, Compound 8, Compound 13, Compound 11, and Compound 58 compared to an ethylene glycol (EG) control. Each solution also contained 300 mM NaCl, 100 mM HEPES, 17.5% (v / v) ethylene glycol, and the pH was adjusted to 7.2. Figure 5A : XRD crystallography (left) and spectrum (right) of compound 10. Figure 5B : XRD crystallography (left) and spectrum (right) of compound 12. Figure 5C : XRD crystallography (left) and spectrum (right) of compound 8. Figure 5D : XRD crystallography (left) and spectrum (right) of compound 13. Figure 5E : XRD crystallography (left) and spectrum (right) of compound 11. Fig. 5F : XRD crystallography (left) and spectrum (right) of compound 58. Figure 5G : EG control XRD crystallography (left) and spectra (right). For the XDR spectra, the intensity is plotted as a function of angle (2θ degrees).

[0054] Figures 6A-6C Two solutions are shown that were quickly frozen, rewarmed, and then frozen again. The control solution contained 22.5% (v / v) ethylene glycol (EG), while the test solution contained 22.5% EG and 5 mg / mL (0.5% (w / v)) Compound 12. Fig. 6A It was shown that during rapid freezing in liquid nitrogen, the solution containing compound 12 vitrified while the control solution was completely frozen. Figure 6B It was shown that during rewarming at 37°C, the solution containing Compound 12 thawed (within two seconds), while the control remained frozen. Figure 6C It is shown that after overnight in a -20°C freezer, the Compound 12 solution remained thawed, unlike the control.

[0055] Figure 7Results of cytotoxicity studies on HEK 293 cells in which compound 12 (squares) or DMSO (circles) were added to the cell culture medium are shown. Samples in which compound 12 or DMSO were not added ("culture medium" (triangles)) were used as controls. Serial dilutions were performed to test different concentrations of compound 12 and DMSO.

[0056] Figure 8 Shown are the results of a cryopreservation experiment on HEK293 cells, comparing solutions containing ethylene glycol (EG) to solutions containing EG and compound 12. Cell viability was measured 12 hours after thawing.

[0057] Fig. 9 Results of cryopreservation experiments on HEK293 cells are shown, comparing solutions containing 5 mg / mL of compound 12 and a mixture of diols, disaccharides, and general buffer to solutions containing VS2E or M22. Cell viability was measured 16 hours after thawing. Cells were vitrified with liquid nitrogen (LN2).

[0058] Fig.10 Shown are the viability data for K562 cells after storage at -20°C for 3 days followed by rewarming for 7 days.

[0059] Fig.11 Shown are survival data for K562 cells 1 day after thawing following 3 days storage at -80°C.

[0060] Fig.12 Shown are survival data for HEK293 cells 1 day after thawing following 3 days storage at -80°C.

[0061] Fig.13 Cytotoxicity data for K562 cells are shown. DETAILED DESCRIPTION

[0062] I. Introduction

[0063] Using cryopreservation to store cells and tissues at low temperatures is crucial for many biological products and applications, but it is still a major problem to successfully fully restore or activate therapeutic cells, tissues, and organs. Cryopreservation is usually performed with cryoprotectants (CPA), which are important chemical additives such as dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), and others. CPA is used to improve the post-thaw survival rate of cryopreserved biological systems by preventing ice crystal nucleation and growth. However, these substances show various levels of cytotoxicity at their effective concentrations, thus limiting the success of cryopreservation, biological storage, and advanced regenerative medicine. The lack of such effective and safe CPAs has led to the widespread use of toxic CPAs. In addition to biological products and applications, preventing ice formation remains a physical and chemical problem faced by a large number of industrial and technical sectors.

[0064] The present invention is based in part on the surprising discovery that N-substituted biomimetic amino acid polymers (peptoids) and peptoid-peptide hybrids have ice crystallization inhibition properties. In addition, the present invention is based in part on the discovery that specific combinations of hydrophobic and polar peptoid monomers can be used to enhance the ice crystallization inhibition properties of the compositions disclosed herein. Provided herein are polymers that reduce or inhibit ice crystal formation below 0°C and at cryogenic temperatures. These polymers can be used to make cryoprotective fluids. Also provided herein are methods for preserving tissues, organs, or cells using cryoprotective fluids, which include using peptoid polymers, peptoid-peptide hybrids, and / or salts thereof as described herein. In addition, beauty care products, deicing products, and frozen foods with antifreeze properties containing peptoid polymers, peptoid-peptide hybrids, and / or salts thereof as described herein are provided. After reading the detailed description, a person of ordinary skill in the art will recognize other advantages that can be derived from the teachings provided herein.

[0065] II. Abbreviations and Definitions

[0066] Abbreviations used herein are conventional unless otherwise defined. The following abbreviations are used to represent specific monomer units of the peptoid polymers: Nsb (2-(sec-butylamino)acetic acid), Nib (2-(isobutylamino)acetic acid), Nbu (2-(butylamino)acetic acid), Npr (2-propylamino)acetic acid), Nip (2-(isopropylamino)acetic acid), Nme (2-(methylamino)acetic acid), Nhp (2-((2-hydroxypropyl)amino)acetic acid), Nhe (2-((2-hydroxyethyl)amino)acetic acid), Ndp (2-((2,3-dihydroxypropyl)amino)acetic acid, Nyp (2-((1-hydroxypropan-2-yl)amino)acetic acid), Nep (2-((1-(4-hydroxyphenyl)ethyl)amino)acetic acid, Ndh (2-((1,3,-dihydroxypropan-2-yl)amino)acetic acid, Nop (2-((3-(2-oxopyrrolidin-1-yl)propyl)amino)acetic acid, Nmo (2-(2-methoxy- acetic acid), Ntf (2-((tetrahydrofuran-2-yl)methylamino)acetic acid), Nff (2-(furan-2-ylmethylamino)acetic acid), Nmb (2-(2-methylbutylamino)acetic acid), Nrh (2-(R)-(2-hydroxypropylamino)acetic acid), Nsh (2-(S)-(2-hydroxypropylamino)acetic acid), N3p (2-(2-(2-(2-methoxyethoxy)ethoxy)ethylamino)acetic acid), ) acetic acid), Nbr ((2-(R)-sec-butylamino)acetic acid), and Nbs ((2-(S)-sec-butylamino)acetic acid). The following abbreviations are used to represent the compounds: DMF (N,N'-dimethylformamide), DIEA (diisopropylethylamine), DIC (N,N'-diisopropylcarbodiimide), ACN (acetonitrile), DCM (dichloromethane), HFIP (hexafluoroisopropanol); Fmoc (9-fluorenylmethoxycarbonyl).

[0067] As used herein, the terms "a," "an," or "the" include not only aspects having one member, but also aspects having more than one member. For example, the singular forms "a," "an," or "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a cell" includes a plurality of such cells, and reference to "the agent" includes one or more agents known to those skilled in the art, and so forth.

[0068] The term "about" as used herein to modify a numerical value represents a limited range around that value. If "X" is that value, "about X" will represent a value of 0.9X to 1.1X, and more preferably, a value of 0.95X to 1.05X. Any reference to "about X" specifically represents at least that value X, 0.95X, 0.96X, 0.97X, 0.98X, 0.99X, 1.01X, 1.02X, 1.03X, 1.04X, and 1.05X. Therefore, "about X" is intended to teach and provide written description support for a claim limitation of, for example, "0.98X".

[0069] The term "alkyl" refers to a straight or branched chain, saturated, aliphatic group having a specified number of carbon atoms. An alkyl group may include any number of carbon atoms, for example, C 1-2 , C 1-3 , C 1-4 , C 1-5 , C 1-6 , C 1-7 , C 1-8 , C 1-9 , C 1-10 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5 , C 4-6 and C 5-6 For example, C 1-6 Alkyl includes but is not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, amyl, isopentyl, hexyl etc.Alkyl can also refer to the alkyl with up to 30 carbon atoms, such as but not limited to heptyl, octyl, nonyl, decyl etc.Alkyl can be substituted or unsubstituted.Alkyl can be optionally selected from halogen, hydroxyl, amino, sulfhydryl, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo (oxo), thioxo (thioxo) and cyano One or more parts replace.

[0070] The term "alkenyl" refers to a straight or branched chain hydrocarbon having at least 2 carbon atoms and at least 1 double bond. Alkenyl groups may include any number of carbon atoms, such as C2, C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 2-7 , C 2-8 , C 2-9 , C 2-10 , C3, C 3-4 , C 3-5 , C 3-6 , C4, C4-5 , C 4-6 , C5, C 5-6 , and C6.Alkenyl can have any suitable number of double bonds, including but not limited to 1, 2, 3, 4, 5 or more. Examples of alkenyl include but are not limited to vinyl (vinyl, ethenyl)), propenyl, isopropenyl, 1-butenyl, 2-butenyl, isobutylenyl, butadienyl, 1-pentenyl, 2-pentenyl, isopentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1,3-hexadienyl, 1,4-hexadienyl, 1,5-hexadienyl, 2,4-hexadienyl or 1,3,5-hexatrienyl.Alkenyl can be substituted or unsubstituted.Alkenyl can be optionally substituted by one or more parts selected from halogen, hydroxyl, amino, sulfhydryl, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio and cyano.

[0071] The term "alkynyl" refers to a straight or branched hydrocarbon having at least 2 carbon atoms and at least one triple bond. Alkynyl groups may include any number of carbon atoms, such as C2, C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 2-7 , C 2-8 , C 2-9 , C 2-10 , C3, C 3-4 , C 3-5 , C 3-6 , C4, C 4-5 , C 4-6 , C5, C 5-6 , and C6. Examples of alkynyl include, but are not limited to, ethynyl, propynyl, 1-butynyl, 2-butynyl, isobutynyl, sec-butynyl, butadiynyl, 1-pentynyl, 2-pentynyl, isopentynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,3-hexadiynyl, 1,4-hexadiynyl, 1,5-hexadiynyl, 2,4-hexadiynyl, or 1,3,5-hexatriynyl. Alkynyl can be substituted or unsubstituted. Alkynyl can be optionally substituted with one or more moieties selected from halogen, hydroxyl, amino, sulfhydryl, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thioxo, and cyano.

[0072] The term "alkylene" refers to a straight or branched, saturated, aliphatic group having a specified number of carbon atoms and connecting at least two other groups, i.e., divalent hydrocarbon groups. The two moieties connected to the alkylene group may be connected to the same atom or to different atoms of the alkylene group. For example, a straight chain alkylene group may be -(CH2) n-divalent radical, wherein n is the number of any suitable carbon atoms. Representational alkylidene includes but is not limited to methylene, ethylene, propylene, isopropylene, butylene, isobutylene, secondary butylene, pentylene and hexylene. Alkylidene can be substituted or unsubstituted. Alkylidene can optionally be selected from one or more parts of halogen, hydroxyl, amino, sulfydryl, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio and cyano.

[0073] The term "alkenylene" refers to an alkenyl group as defined above that is connected to at least two other groups, i.e. a divalent hydrocarbon group. The two parts connected to the alkenylene group can be connected to the same atom or different atoms of the alkenylene group. The alkenylene group includes but is not limited to vinylene, propenylene, isopropenylene, butenylene, isobutenylene, secondary butenylene, pentenylene and hexenylene. The alkenylene group can be substituted or unsubstituted. The alkenylene group can optionally be selected from one or more parts of halogen, hydroxyl, amino, sulfydryl, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio and cyano.

[0074] The term "alkynylene" refers to an alkynyl group as defined above that is connected to at least two other groups, i.e., a divalent hydrocarbon group. The two parts connected to the alkynylene group can be connected to the same atom or different atoms of the alkynylene group. Alkynylidene includes but is not limited to ethynylene, propynylene, isopropynylene, butynylene, secondary butynylene, pentynylene and hexynylene. Alkynylidene can be substituted or unsubstituted. Alkynylidene can be optionally substituted by one or more parts selected from halogen, hydroxyl, amino, sulfhydryl, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio and cyano.

[0075] The term "halogen" or "halo" refers to fluorine, chlorine, bromine and iodine.

[0076] The term "amine" or "amino" refers to -N(R)2 groups, wherein the R groups can be hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, etc. The R groups can be the same or different. The amino group can be a primary amino group (each R is hydrogen), a secondary amino group (one R is hydrogen) or a tertiary amino group (each R is not hydrogen). Alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl can be optionally substituted by one or more parts selected from halogen, hydroxyl, amino, sulfhydryl, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio and cyano.

[0077] The term "hydroxyl" or "hydroxy" refers to an -OH group. The hydroxyl group may be at any suitable carbon atom.

[0078] The term "mercapto" refers to a -SH group. The mercapto group may be at any suitable carbon atom.

[0079] The term "oxo" refers to a double bond O group (=O, -C(O)-). The oxo group may be at any suitable carbon atom.

[0080] The term "thio" refers to a double bond S group (=S). The thio group may be at any suitable carbon atom.

[0081] The term "nitro" refers to a -NO2 group. The nitro group may be at any suitable carbon atom.

[0082] The term "carboxyl" refers to a carboxylic acid group of the formula -C(O)OH or -CO2H.

[0083] The term "cycloalkyl" refers to a saturated or partially unsaturated, monocyclic, fused bicyclic or bridged polycyclic ring assembly containing 3 to 12 ring atoms or the specified number of atoms. Cycloalkyl may include any number of carbon atoms, such as C 3-6 , C 4-6 , C 5-6 , C 3-8 , C 4-8 , C 5-8 , C 6-8 , C 3-9 , C 3-10 , C 3-11 , and C 3-12 . Saturated monocyclic cycloalkyl rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. Saturated bicyclic and polycyclic cycloalkyl rings include, for example, norbornane, [2.2.2]bicyclooctane, decalin, and adamantane. Cycloalkyl groups may also be partially unsaturated, having one or more double or triple bonds in the ring. Representative partially unsaturated cycloalkyl groups include, but are not limited to, cyclobutene, cyclopentene, cyclohexene, cyclohexadiene (1,3- and 1,4-isomers), cycloheptene, cycloheptadiene, cyclooctene, cyclooctadiene (1,3-, 1,4- and 1,5-isomers), norbornene, and norbornadiene. When the cycloalkyl group is a saturated monocyclic C 3-8 When the cycloalkyl group is a saturated monocyclic C 3-6 When the cycloalkyl group is a cycloalkyl group, exemplary groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. The cycloalkyl group may be substituted or unsubstituted. The cycloalkyl group may be optionally substituted by one or more moieties selected from alkyl, alkenyl, alkynyl, halogen, hydroxyl, amino, alkylamino, alkoxy, haloalkyl, carboxyl, amido, sulfhydryl, nitro, oxo, thio, and cyano. For example, the cycloalkyl group may be C 1-6 The radical is substituted with an alkyl radical or an oxo radical (═O).

[0084] The term "heterocycloalkyl" refers to a saturated ring system having 3 to 12 ring members and 1 to 4 heteroatoms of N, O or S. Additional heteroatoms are also useful, including but not limited to B, Al, Si and P. Heteroatoms may also be oxidized to form moieties including but not limited to -S(O)- and -S(O)2-. Heterocycloalkyl may include any number of ring atoms, such as 3 to 6, 4 to 6, 5 to 6, 3 to 8, 4 to 8, 5 to 8, 6 to 8, 3 to 9, 3 to 10, 3 to 11 or 3 to 12 ring members. Any suitable number of heteroatoms may be included in the heterocycloalkyl, such as 1, 2, 3 or 4, or 1 to 2, 1 to 3, 1 to 4, 2 to 3, 2 to 4 or 3 to 4. The heterocycloalkyl group may include groups such as aziridine, azetidine, pyrrolidine, piperidine, azepane, azocane, quinuclidine, pyrazolidine, imidazolidine, piperazine (1,2-, 1,3- and 1,4-isomers), oxirane, oxetane, tetrahydrofuran, oxane (tetrahydropyran), hexene oxide, thiirane, thietane, thiolane (tetrahydrothiophene), thiane (tetrahydrothiopyrane), thiazolidine ... etrahydrothiopyran), oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, dioxolane, dithiolane, morpholine, thiomorpholine, dioxane or dithiane. Heterocycloalkyl can also be fused with an aromatic or non-aromatic ring system to form a member including but not limited to dihydroindole. Heterocycloalkyl can be substituted or unsubstituted. Heterocycloalkyl can be optionally substituted by one or more parts selected from alkyl, alkenyl, alkynyl, halogen, hydroxyl, amino, sulfhydryl, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio, and cyano. Wherein, for example, heterocycloalkyl can be C 1-6 The radical is substituted with an alkyl radical or an oxo radical (═O).

[0085] The heterocycloalkyl group may be attached at any position on the ring. For example, the aziridine can be 1- or 2-aziridine, the azetidine can be 1- or 2-azetidine, the pyrrolidine can be 1-, 2-, or 3-pyrrolidine, the piperidine can be 1-, 2-, 3-, or 4-piperidine, the pyrazolidine can be 1-, 2-, 3-, or 4-pyrazolidine, the imidazolidine can be 1-, 2-, 3-, or 4-imidazolidine, the piperazine can be 1-, 2-, 3-, or 4-piperazine, the tetrahydrofuran can be 1- or 2-tetrahydrofuran, the oxazolidine can be 2-, 3-, 4-, or 5-oxazolidine, the isoxazolidine can be 2-, 3-, 4-, or 5-isoxazolidine, the thiazolidine can be 2-, 3-, 4-, or 5-thiazolidine, the isothiazolidine can be 2-, 3-, 4-, or 5-isothiazolidine, and the morpholine can be 2-, 3-, or 4-morpholine.

[0086] When heterocycloalkyl includes 3 to 8 ring members and 1 to 3 heteroatoms, representative members include but are not limited to pyrrolidine, piperidine, tetrahydrofuran, ethylene oxide, tetrahydrothiophene, cyclopentyl sulfide, pyrazolidine, imidazolidine, piperazine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine, morpholine, thiomorpholine, dioxane and dithiane.Heterocycloalkyl can also form a ring with 5 to 6 ring members and 1 to 2 heteroatoms, and representative members include but are not limited to pyrrolidine, piperidine, tetrahydrofuran, tetrahydrothiophene, pyrazolidine, imidazolidine, piperazine, oxazolidine, isoxazolidine, thiazolidine, isothiazolidine and morpholine.

[0087] The term "aryl" refers to an aromatic ring system with any suitable number of ring atoms and any suitable number of rings. The aryl group can include any suitable number of ring atoms, such as 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 ring atoms, and 6 to 10, 6 to 12 or 6 to 14 ring members. The aryl group can be a monocyclic ring, fused to form a bicyclic or tricyclic group, or connected by a bond to form a biaryl group. Representative aryl groups include phenyl, naphthyl and biphenyl. Other aryl groups include benzyl with a methylene linking group. Some aryl groups have 6 to 12 ring members, such as phenyl, naphthyl or biphenyl. Other aryl groups have 6 to 10 ring members, such as phenyl or naphthyl. Some other aryl groups have 6 ring members, such as phenyl. The aryl group can be substituted or unsubstituted. The aryl group can be optionally substituted with one or more moieties selected from alkyl, alkenyl, alkynyl, halogen, hydroxy, amino, mercapto, alkylamino, alkoxy, haloalkyl, carboxy, amido, nitro, oxo, thio, and cyano.

[0088] The term "heteroaryl" refers to a monocyclic or fused bicyclic or tricyclic aromatic ring set containing 5 to 16 ring atoms, wherein 1 to 5 ring atoms are heteroatoms such as N, O or S. Other heteroatoms are also useful, including but not limited to B, Al, Si and P. Heteroatoms can also be oxidized to form parts including but not limited to-S(O)- and-S(O)2-. Heteroaryl can include any number of ring atoms, such as 3 to 6, 4 to 6, 5 to 6, 3 to 8, 4 to 8, 5 to 8, 6 to 8, 3 to 9, 3 to 10, 3 to 11 or 3 to 12 ring members. Any suitable number of heteroatoms can be included in the heteroaryl, such as 1, 2, 3, 4 or 5, or 1 to 2, 1 to 3, 1 to 4, 1 to 5, 2 to 3, 2 to 4, 2 to 5, 3 to 4 or 3 to 5. Heteroaryl can have 5 to 8 ring members and 1 to 4 heteroatoms, or 5 to 8 ring members and 1 to 3 heteroatoms, or 5 to 6 ring members and 1 to 4 heteroatoms, or 5 to 6 ring members and 1 to 3 heteroatoms.Heteroaryl can include the following groups: for example pyrrole, pyridine, imidazole, pyrazole, triazole, tetrazole, pyrazine, pyrimidine, pyridazine, triazine (1,2,3-, 1,2,4- and 1,3,5-isomers), thiophene, furan, thiazole, isothiazole, oxazole and isoxazole.Heteroaryl can also be fused to aromatic ring systems such as benzene rings to form members including but not limited to the following: benzopyrrole such as indole and isoindole, benzopyridine such as quinoline and isoquinoline, benzopyrazine (quinoxaline), benzopyrimidine (quinazoline), benzopyridazine such as phthalazine and cinnoline, benzothiophene and benzofuran. Other heteroaryls include heteroaryl rings connected by bonds, such as bipyridine. Heteroaryl can be substituted or unsubstituted. Heteroaryl can be optionally substituted with one or more moieties selected from alkyl, alkenyl, alkynyl, halogen, hydroxyl, amino, sulfhydryl, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio and cyano.

[0089] The heteroaryl group can be attached at any position on the ring. For example, pyrrole includes 1-, 2-, and 3-pyrrole, pyridine includes 2-, 3-, and 4-pyridine, imidazole includes 1-, 2-, 4-, and 5-imidazole, pyrazole includes 1-, 3-, 4-, and 5-pyrazole, triazole includes 1-, 4-, and 5-triazole, tetrazole includes 1- and 5-tetrazole, pyrimidine includes 2-, 4-, 5-, and 6-pyrimidine, pyridazine includes 3- and 4-pyridazine, 1,2,3-triazine includes 4- and 5-triazine, 1,2,4-triazine includes 3-, 5-, and 6-triazine, 1,3,5-triazine includes 2-triazine, thiophene includes 2- and 3-thiophene, furan includes 2- and 3-thiophene, including 2- and 3-furan, thiazole including 2-, 4-, and 5-thiazole, isothiazole including 3-, 4-, and 5-isothiazole, oxazole including 2-, 4-, and 5-oxazole, isoxazole including 3-, 4-, and 5-isoxazole, indole including 1-, 2-, and 3-indole, isoindole including 1- and 2-isoindole, quinoline including 2-, 3-, and 4-quinoline, isoquinoline including 1-, 3-, and 4-isoquinoline, quinazoline including 2- and 4-quinazoline, cinnoline including 3- and 4-cinnoline, benzothiophene including 2- and 3-benzothiophene, and benzofuran including 2- and 3-benzofuran.

[0090] Some heteroaryl groups include those having 5 to 10 ring members and 1 to 3 ring atoms including N, O, or S, such as pyrrole, pyridine, imidazole, pyrazole, triazole, pyrazine, pyrimidine, pyridazine, triazine (1,2,3-, 1,2,4- and 1,3,5-isomers), thiophene, furan, thiazole, isothiazole, oxazole, isoxazole, indole, isoindole, quinoline, isoquinoline, quinoxaline, quinazoline, phthalazine, cinnoline, benzothiophene, and benzofuran. Other heteroaryl groups include those having 5 to 8 ring members and 1 to 3 heteroatoms, such as pyrrole, pyridine, imidazole, pyrazole, triazole, pyrazine, pyrimidine, pyridazine, triazine (1,2,3-, 1,2,4- and 1,3,5-isomers), thiophene, furan, thiazole, isothiazole, oxazole, and isoxazole. Some other heteroaryl groups include those with 9 to 12 ring members and 1 to 3 heteroatoms, such as indole, isoindole, quinoline, isoquinoline, quinoxaline, quinazoline, phthalazine, cinnoline, benzothiophene, benzofuran, and bipyridine. Other heteroaryl groups include those with 5 to 6 ring members and 1 to 2 ring atoms including N, O, or S, such as pyrrole, pyridine, imidazole, pyrazole, pyrazine, pyrimidine, pyridazine, thiophene, furan, thiazole, isothiazole, oxazole, and isoxazole.

[0091] The term "(cycloalkyl)alkyl" refers to a group having an alkyl component and a cycloalkyl component, wherein the alkyl component connects the cycloalkyl component to the point of attachment. The alkyl component is defined as above, except that the alkyl component is at least divalent, and an alkylene group is connected to the cycloalkyl component and the point of attachment. The alkyl component may include any number of carbon atoms, such as C 1-6, C 1-2 , C 1-3 , C 1-4 , C 1-5 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5 , C 4-6 and C 5-6 The cycloalkyl component is as defined above. Exemplary (cycloalkyl)alkyl groups include, but are not limited to, methyl-cyclopropyl, methyl-cyclobutyl, methyl-cyclopentyl, and methyl-cyclohexyl.

[0092] The term "(heterocycloalkyl)alkyl" refers to a group having an alkyl component and a heterocycloalkyl component, wherein the alkyl component connects the heterocycloalkyl component to the point of attachment. The alkyl component is as defined above, except that the alkyl component is at least divalent, and an alkylene group is connected to the heterocycloalkyl component and the point of attachment. The alkyl component may include any number of carbon atoms, such as C 0-6 , C 1-2 , C 1-3 , C 1-4 , C 1-5 , C 1-6 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5 , C 4-6 and C 5-6 The heterocycloalkyl component is as defined above. The (heterocycloalkyl)alkyl group may be substituted or unsubstituted.

[0093] The term "aralkyl" refers to a group having an alkyl component and an aryl component, wherein the alkyl component connects the aryl component to the point of attachment. The alkyl component is as defined above, except that the alkyl component is at least divalent, and the alkylene group is connected to the aryl component and the point of attachment. The alkyl component may include any number of carbon atoms, such as C 0-6 , C 1-2 , C 1-3 , C 1-4 , C 1-5 , C 1-6 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5, C 4-6 and C 5-6 The aryl component is as defined above. Examples of aralkyl groups include, but are not limited to, benzyl and ethyl-benzene. Aralkyl groups may be substituted or unsubstituted.

[0094] The term "heteroaralkyl" refers to a group having an alkyl component and a heteroaryl component, wherein the alkyl component connects the heteroaryl component to the point of attachment. The alkyl component is as defined above, except that the alkyl component is at least divalent, and the alkylene group is connected to the heteroaryl component and the point of attachment. The alkyl component may include any number of carbon atoms, such as C 0-6 , C 1-2 , C 1-3 , C 1-4 , C 1-5 , C 1-6 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5 , C 4-6 and C 5-6 The heteroaryl component is as defined above. The heteroaralkyl group may be substituted or unsubstituted.

[0095] The term "carboxyalkyl" refers to a carboxyl group attached to an alkyl group as described above, and generally has the formula -C 1-8 Alkyl-C(O)OH. Any suitable alkyl chain is useful. The carboxyalkyl group may be optionally substituted with one or more moieties selected from halogen, hydroxy, amino, thiol, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio, and cyano.

[0096] The term "acyl" refers to an alkyl group containing an oxygen-substituted carbon at the point of attachment (-C(O)-C 1-8 The acyl group may be optionally substituted with one or more moieties selected from halogen, hydroxy, amino, mercapto, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio, and cyano.

[0097] The term "hydroxyalkyl" refers to an alkyl group as defined above in which at least one hydrogen atom is replaced by a hydroxy group. With respect to the alkyl group, the hydroxyalkyl group may have any suitable number of carbon atoms, for example C 1-6. Exemplary hydroxyalkyl groups include, but are not limited to, hydroxymethyl, hydroxyethyl (wherein the hydroxyl group is at the 1- or 2-position), hydroxypropyl (wherein the hydroxyl group is at the 1-, 2- or 3-position), hydroxybutyl (wherein the hydroxyl group is at the 1-, 2-, 3- or 4-position), hydroxypentyl (wherein the hydroxyl group is at the 1-, 2-, 3-, 4- or 5-position), hydroxyhexyl (wherein the hydroxyl group is at the 1-, 2-, 3-, 4-, 5- or 6-position), and 1,2-dihydroxyethyl, etc. The hydroxyalkyl group may be optionally substituted with one or more moieties selected from halogen, sulfhydryl, amino, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio, and cyano. It will be appreciated by those skilled in the art that other hydroxyalkyl groups are useful in the present invention.

[0098] The term "alkoxy" refers to an alkyl group having at least one bridging oxygen atom. The bridging oxygen atom may be at any position within the alkyl chain (alkyl-O-alkyl), or the bridging oxygen atom may connect the alkyl group to the point of attachment (alkyl-O-). In some embodiments, the bridging oxygen atom does not exist as a terminal hydroxyl group (i.e., -OH). In some examples, the alkoxy group contains 1, 2, 3, 4, or 5 bridging oxygen atoms. With respect to alkyl, the alkoxy group may have any suitable number of carbon atoms, such as C 1-2 , C 1-4 , and C 1-6 Alkoxy groups include, for example, methoxy, ethoxy, propoxy, isopropoxy, methyloxy-ethyloxy-ethyl (C1-O-C2-O-C2-), etc. An example of an alkoxy group is polyethylene glycol (PEG), wherein the polyethylene glycol chain may include between 2 and 20 ethylene glycol monomers. The alkoxy group may optionally be substituted with one or more moieties selected from halogen, hydroxyl, amino, sulfhydryl, alkylamino, haloalkyl, carboxyl, amido, nitro, oxo, thio, and cyano. The alkoxy group may be substituted or unsubstituted.

[0099] The term "alkylamino" refers to an alkyl group as defined above having one or more amino groups. The amino group may be a primary amino group, a secondary amino group or a tertiary amino group. Alkylamino groups that can be used in the present invention include, but are not limited to, ethylamine, propylamine, isopropylamine, ethylenediamine and ethanolamine. The amino group may be connected to the point of attachment, with the remainder of the compound being at any position of the alkyl group, or at least two carbon atoms of the alkyl group being connected together. The alkylamino group may optionally be substituted with one or more moieties selected from halogen, hydroxyl, sulfhydryl, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio and cyano groups. It will be appreciated by those skilled in the art that other alkylamino groups are useful in the present invention.

[0100] The term "alkylthio" refers to an alkyl group as defined above having one or more sulfhydryl groups. Alkylthio groups that can be used in the present invention include, but are not limited to, ethyl mercapto, propyl mercapto, and isopropyl mercapto. The mercapto group can connect the alkylthio group to a point of attachment, wherein the remainder of the compound is at any position of the alkyl group, or at least two carbon atoms of the alkyl group can be connected together. The alkylthio group can optionally be substituted with one or more parts selected from halogen, hydroxyl, amino, alkylamino, alkoxy, haloalkyl, carboxyl, amido, nitro, oxo, thio, and cyano. It will be appreciated by those skilled in the art that other alkylthio groups are useful in the present invention.

[0101] The term "oxyethyl" refers to a divalent group having the formula -OCH2CH2-.

[0102] The term "wavy line" indicates the point of attachment of a substituent to the rest of the molecule. When the wavy line is not described as being specifically attached to a particular ring atom, the point of attachment may be any suitable atom of the substituent. For example, the wavy line in the following structure:

[0103]

[0104] Any substitutable atom is intended to be included as a point of attachment.

[0105] The term "regenerative medicine" refers to the branch of medicine that deals with the process of replacing, altering, or regenerating human cells, tissues, or organs to restore or establish normal function. In some embodiments, regenerative medicine involves growing tissues and organs in the laboratory and safely transplanting them when the body cannot heal itself.

[0106] The term "bioengineered tissue" refers to one or more synthetically produced cells, tissues, or organs produced for the purpose of regenerative medicine. In some embodiments, bioengineered tissue refers to cells, tissues, or organs developed in a laboratory. In some embodiments, bioengineered tissue refers to laboratory-derived heart, liver, lung, kidney, pancreas, intestine, thymus, cornea, stem cells (e.g., human pluripotent stem cells, hematopoietic stem cells), lymphocytes, granulocytes, immune system cells, bone cells, primary cells, organoids, embryonic cells, genitourinary cells (e.g., sperm cells, oocytes, corpus cavernosum cells (e.g., corpus cavernosum smooth muscle cells, corpus cavernosum epithelial cells), bladder cells, urethral cells, ureteral cells, kidney cells, testicular cells), platelets, neural cells, or combinations thereof.

[0107] The term "cryoprotectant" refers to a solution used to reduce or prevent freezing damage caused by ice crystal formation. In some embodiments, the cryoprotectant comprises one or more peptoid polymers and / or salts thereof as described herein. In other embodiments, the cryoprotectant comprises one or more peptoid polymers and / or salts thereof as described herein, and one or more peptoid-peptide hybrids and / or salts thereof as described herein. In some embodiments, the cryoprotectant protects biological samples from freezing damage. In some embodiments, the cryoprotectant protects non-biological samples from ice crystal formation. In some embodiments, the cryoprotectant preserves the biological sample for a longer period of time than when the biological sample is not exposed to a reduced temperature.

[0108] The terms "vitrify" and "vitrification" refer to the conversion of a substance into a glassy state (i.e., a non-crystalline amorphous solid). In the case of water, vitrification refers to the conversion of water into a glassy state without the formation of ice crystals, as opposed to ordinary freezing, which results in the formation of ice crystals. Vitrification is usually achieved by very rapid cooling and / or the introduction of agents that inhibit the formation of ice crystals. On the other hand, "devitrify" and "devitrification" refer to the process of crystallization in a previously crystalline (amorphous) glassy state. In the case of ice water, devitrification can mean the formation of ice crystals as the previously non-crystalline amorphous solid undergoes melting.

[0109] The term "peptoid monomer" refers to a peptide according to formula (I) comprising a side chain R 1 and R 2 The molecule:

[0110]

[0111] and includes stereoisomers and tautomers thereof. In some embodiments, R 1 Selected from the group consisting of: H, hydroxy, optionally substituted C 1-18 Alkyl, optionally substituted C 2-18 Alkenyl, optionally substituted C 2-18 Alkynyl, optionally substituted C 1-18 Hydroxyalkyl, optionally substituted alkoxy, optionally substituted C 1-18 Alkylamino, optionally substituted C 1-18 alkylthio, optionally substituted carboxyalkyl, C 3-10 Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, (C 3-10 cycloalkyl)alkyl, (heterocycloalkyl)alkyl, aralkyl, and heteroaralkyl, wherein any of cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is optionally substituted by one or more R 3 In certain embodiments, R3 Selected from halogen, oxo, thio, -OH, -SH, sulfonamide, amino, C 1-8 Alkyl, C 1-8 Hydroxyalkyl, C 1-8 Alkylamino, and C 1-8 In addition, R 1 The side chain of any of the following amino acids may be included: alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), arginine (Arg), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), or tyrosine (Tyr).

[0112] R 1 and R 2 In some embodiments, R 2 is H. In other embodiments, R 2 Selected from the group consisting of: hydroxy, optionally substituted C 1-18 Alkyl, optionally substituted C 2-18 Alkenyl, optionally substituted C 2-18 Alkynyl, optionally substituted C 1-18 Hydroxyalkyl, optionally substituted alkoxy, optionally substituted C 1-18 Alkylamino, optionally substituted C 1-18 alkylthio, optionally substituted carboxyalkyl, C 3-10 Cycloalkyl, heterocycloalkyl, aryl, heteroaryl, (C 3-10 cycloalkyl)alkyl, (heterocycloalkyl)alkyl, aralkyl, and heteroaralkyl, wherein any of cycloalkyl, heterocycloalkyl, aryl, and heteroaryl is optionally substituted by one or more R 3 In certain embodiments, R 3 Selected from halogen, oxo, thio, -OH, -SH, sulfonamide, amino, C 1-8 Alkyl, C 1-8 Hydroxyalkyl, C 1-8 Alkylamino, and C 1-8 In addition, R 2The side chain of any of the following amino acids may be included: alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), arginine (Arg), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), or tyrosine (Tyr).

[0113] In some embodiments, R 1 and / or R 2 Includes structure:

[0114]

[0115] Where R 4 Choose from H, C 1-8 The group consisting of alkyl, hydroxy, sulfonamide, mercapto, nitro, amine, oxo, and thio.

[0116] In some embodiments, the peptoid monomer is selected from the group of peptoid monomers described in Table 1. Those skilled in the art will understand that the boundaries of the present invention are not limited to the monomers listed in Table 1, and any useful N-substituted substituents can be used as N-substituted peptoid monomers.

[0117] X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 In some embodiments, X is acetyl. In some embodiments, Y is carboxyl. Alternatively, X and Y together form a covalent bond. When a peptoid monomer is present as part of a polymer, X and / or Y may represent a covalent bond to another peptoid monomer.

[0118] Whenever stereochemistry is not indicated for any peptoid monomer herein, any stereoisomer may be used. In some embodiments, a mixture of two stereoisomers is selected. In embodiments comprising a mixture of stereoisomers, the ratio of the R to S stereoisomers of the monomers in the peptoid polymer may range from about 95:5 to about 90:10, from about 90:10 to about 85:15, from about 85:15 to about 80:20, from about 80:20 to about 75:25, from about 75:25 to about 70:30, from about 70:30 to about 65:35, from about 65:35 to about 60:40, from about 60:40 to about 65:50. From about 55:45 to about 50:50, from about 50:50 to about 45:55, from about 45:55 to about 40:60, from about 40:60 to about 35:65, from about 35:65 to about 30:70, from about 30:70 to about 25:75, from about 25:75 to about 20:80, from about 20:80 to about 15:85, from about 15:85 to about 10:90, or from about 10:90 to about 5:95. In some embodiments, only the R stereoisomer of the monomer is selected. In some embodiments, only the S stereoisomer of the monomer is selected.

[0119] Whenever a specific stereochemistry is shown with a wedge or dashed line, the monomer is substantially free of other stereoisomers. In some embodiments, substantially free means at least 70% pure. In some embodiments, substantially free means at least 80% pure. In some embodiments, substantially free means at least 90% pure. In some embodiments, substantially free means at least 95% pure. In some embodiments, substantially free means at least 99% pure. In some embodiments, substantially free means at least 99.9% pure.

[0120] In some embodiments, one or more R 1 and / or R 2 The side chain has 1a The structure:

[0121]

[0122] In some embodiments, each R 1a The groups are independently selected from In some embodiments, a mixture of two stereoisomers is selected. In some embodiments, only the R stereoisomer of a monomer is selected. In some embodiments, only the S stereoisomer of the monomer is selected.

[0123] In some embodiments, one or more R 1 and / or R 2 The side chain has 1b The structure:

[0124]

[0125] In some embodiments, each R 1b The groups are independently selected from In some embodiments, a mixture of two stereoisomers is selected. In some embodiments, only the R stereoisomer of a monomer is selected. In some embodiments, only the S stereoisomer of the monomer is selected.

[0126] In some embodiments, one or more R 1 and / or R 2 The side chain has 1c The structure:

[0127]

[0128] In some embodiments, each R 1c The groups are independently selected from In some embodiments, a mixture of two stereoisomers is selected. In some embodiments, only the R stereoisomer of a monomer is selected. In some embodiments, only the S stereoisomer of the monomer is selected.

[0129] In some embodiments, one or more R 1 and / or R 2 The side chain has 1d The structure:

[0130]

[0131] In some embodiments, each R 1d The groups are independently selected from In some embodiments, a mixture of two stereoisomers is selected. In some embodiments, only the R stereoisomer of a monomer is selected. In some embodiments, only the S stereoisomer of the monomer is selected.

[0132] In some embodiments, one or more R 1 and / or R 2 The side chain has 1e The structure:

[0133]

[0134] In some embodiments, each R 1e The groups are independently selected from In some embodiments, a mixture of two stereoisomers is selected. In some embodiments, only the R stereoisomer of a monomer is selected. In some embodiments, only the S stereoisomer of the monomer is selected.

[0135] The terms "polar peptoid monomer" and "peptoid monomer with polar side chains" are used interchangeably to refer to a peptide in which the substituent "R 1 " is a polar side chain, or R 1 and the substituent "R 2 " Both are peptoid monomers with polar side chains. Typically, polar side chains include hydroxyl groups and / or atoms (e.g., sulfur, nitrogen, oxygen) that can participate in hydrogen bonding. In some examples, polar side chains include atoms or groups that are more hydrophobic in nature relative to polarity (e.g., aromatic rings). In these examples, the side chains also include atoms or groups that make the entire side chain more polar relative to hydrophobicity. As a non-limiting example, the polar side chain may include an aromatic ring to which one or more hydroxyl groups are attached.

[0136] Examples of polar peptoid monomers include, but are not limited to,

[0137]

[0138]

[0139] wherein the subscript m is the number of repeating units and is between 1 and 10.

[0140] In some embodiments, the peptoid polymer of the present invention or a salt thereof includes one or more polar peptoid monomers selected from the group consisting of Nop, Nhp, Nhe, Ndp, Nyp, Nep, Ndh, and combinations thereof.

[0141] The terms "hydrophobic peptoid monomer" and "peptoid monomer having a hydrophobic side chain" are used interchangeably to refer to a peptoid monomer in which the substituent "R 1 " is a hydrophobic side chain (e.g., non-polar), or R 1 and the substituent "R 2 " Both are peptoid monomers with hydrophobic side chains. Typically, the hydrophobic side chains include unsubstituted alkyl groups, unsubstituted cycloalkyl groups, or unsubstituted aromatic groups.

[0142] Examples of hydrophobic peptoid monomers include, but are not limited to

[0143]

[0144] wherein the subscript m is the number of repeating units and is between 1 and 10.

[0145] In some embodiments, the peptoid polymer or salt thereof of the present invention includes one or more hydrophobic peptoid monomers selected from the group consisting of Nsb, Nib, Nbu, Npr, Nip, Nme, and combinations thereof.

[0146] The term "peptoid polymer" or "peptoid" refers to a polyamide of between about 2 and 1,000 (e.g., between about 2 and 1,000, 2 and 950, 2 and 900, 2 and 850, 2 and 800, 2 and 750, 2 and 700, 2 and 650, 2 and 600, 2 and 550, 2 and 500, 2 and 450, 2 and 400, 2 and 350, 2 and 300, 2 and 250, 2 and 200, 2 and 150, 2 and 100, 2 and 90, 2 and 80, 2 and 70, 2 and 60, 2 and 50, 2 and 40, 2 and 30, 2 and 20, 2 and 10, 2 and 9, 2 and 8, 2 and 7, 2 and 6, 2 and 5, 2 and 4, or 2 and 3) peptoid monomers. In a specific example, a peptoid is a synthetic analog of a peptide in which the side chain originally attached to the α-carbon atom is instead attached to the amide nitrogen atom. Peptoids are synthetic polymers of controlled sequence and length that can be prepared by automated solid phase organic synthesis to include a variety of side chains with different chemical functionalities. The term includes free amine forms as well as salt forms.

[0147] The term "peptoid-peptide hybrid" refers to an oligomer comprising both a peptoid monomer unit and an alpha amino acid (ie, a peptide unit). The term includes the free amine form as well as the salt form.

[0148] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to a polymer of amino acid residues, or a collection of multiple polymers of amino acid residues.

[0149] The term "amino acid" includes, but is not limited to, naturally occurring α-amino acids and stereoisomers thereof. A "stereoisomer" of an amino acid refers to a mirror image isomer of an amino acid, such as an L-amino acid or a D-amino acid. For example, a stereoisomer of a naturally occurring amino acid refers to a mirror image isomer of a naturally occurring amino acid (i.e., a D-amino acid).

[0150] Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified (e.g., hydroxyproline, γ-carboxyglutamate, and O-phosphoserine). Naturally occurring α-amino acids include, but are not limited to, alanine (Ala), cysteine ​​(Cys), aspartic acid (Asp), glutamic acid (Glu), phenylalanine (Phe), glycine (Gly), histidine (His), isoleucine (Ile), arginine (Arg), lysine (Lys), leucine (Leu), methionine (Met), asparagine (Asn), proline (Pro), glutamine (Gln), serine (Ser), threonine (Thr), valine (Val), tryptophan (Trp), tyrosine (Tyr), and combinations thereof. Stereoisomers of naturally occurring α-amino acids include, but are not limited to, D-alanine (D-Ala), D-cysteine ​​(D-Cys), D-aspartic acid (D-Asp), D-glutamic acid (D-Glu), D-phenylalanine (D-Phe), D-histidine (D-His), D-isoleucine (D-Ile), D-arginine (D-Arg), D-lysine (D-Lys), D-leucine (D-Leu), D-methionine (D-Met), D-asparagine (D-Asn), D-proline (D-Pro), D-glutamine (D-Gln), D-serine (D-Ser), D-threonine (D-Thr), D-valine (D-Val), D-tryptophan (D-Trp), D-tyrosine (D-Tyr), and combinations thereof.

[0151] Amino acids may be referred to herein by their generally recognized three-letter symbols or by the single-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. For example, L-amino acids may be represented herein by their generally recognized three-letter symbols (e.g., Arg represents L-arginine) or by uppercase single-letter amino acid symbols (e.g., R represents L-arginine). D-amino acids may be represented herein by their generally recognized three-letter symbols (e.g., D-Arg represents D-arginine) or by lowercase single-letter amino acid symbols (e.g., r represents D-arginine).

[0152] III. Detailed Description of the Implementation Scheme

[0153] A. Peptide polymers

[0154] In one aspect, the present invention provides a peptoid polymer or a salt thereof comprising subunits, wherein the subunits comprise one or more first hydrophobic peptoid monomers H and one or more first polar peptoid monomers P, which are arranged such that the peptoid polymer has a sequence [H a Pb ] n or [P b H a ] n , wherein the subscript a represents the number of consecutive first hydrophobic peptoid monomers in the subunit, the subscript b represents the number of consecutive first polar peptoid monomers in the subunit, and the subscript n represents the number of subunits in the peptoid polymer. In some embodiments, a is between 1 and 10 (e.g., a is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In other embodiments, b is between 1 and 10 (e.g., b is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some instances, a is between 1 and 5. In other instances, b is between 1 and 5. In specific instances, a is between 1 and 3 and b is between 1 and 3.

[0155] In some embodiments, the subunit further comprises a second hydrophobic peptoid monomer and / or a second polar peptoid monomer, such that the peptoid polymer has the sequence [H a P b H c P d ] n or [P b H a P d H c ] n , wherein the subscript c represents the number of consecutive second hydrophobic peptoid monomers in the subunit, and the subscript d represents the number of consecutive second polar peptoid monomers in the subunit. In some embodiments, c is between 0 and 10 (e.g., c is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In other embodiments, d is between 0 and 10 (e.g., d is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In specific embodiments, c and d are not both 0. In some examples, c is between 0 and 5. In other examples, d is between 0 and 5.

[0156] As a non-limiting example, the subunit of the peptoid polymer of the present invention may include the sequence HP, PH, HHPP, PPHH, HPHP, PHPH, HPPH, PHHP, HHP, PHH, HPP, PPH, HPPP, PPPH, HHPPHH, PPHHPP, HHHPPP, PPPHHH, HHHP, PHHH, HHHPPPHHH, or PPPHHHPPP. When a is 1 and b is 1, the subunit may include the sequence HP or PH. When a is 2 and b is 2, the subunit may include the sequence HHPP or PPHH. When a is 1 and b is 2, the subunit may include the sequence HPP or PPH. When a is 2 and b is 1, the subunit may include the sequence HHP or PHH. When a is 1 and b is 3, the subunit may include the sequence HPPP or PPPH. When a is 3 and b is 1, the subunit may include the sequence HHHP or PHHH. When a is 3 and b is 3, the subunit may comprise the sequence HHHPPP or PPPHHH.

[0157] As a further non-limiting example, when a, b, c, and d are 1, the subunit may include the sequence HPHP or PHPH, although these sequences may also be represented by the formulas [H1P1]2 and [P1H1]2, respectively, where n is 2. When a is 1, b is 2, c is 1, and d is 0, the subunit may include the sequence HPPH. When a is 2, b is 1, c is 0, and d is 1, the subunit may include the sequence PHHP (i.e., P1H2P1HO). When a, b, and c are 2 and d is 0, the subunit may include the sequence HHPPHH. When a, b, and d are 2 and c is 0, the subunit may include the sequence PPHHPP (i.e., P2H2P2HO). When a, b, and c are 3 and d is 0, the subunit may include the sequence HHHPPPHHH. When a, b, and d are 3 and c is 0, the subunit may include the sequence PPPHHHPPP (i.e., P3H3P3HO).

[0158] When there is more than one hydrophobic peptoid monomer in the peptoid polymer or its salt, all of the hydrophobic peptoid monomers can be the same, they can all be different, or a combination thereof. Similarly, when there is more than one polar peptoid monomer in the peptoid polymer or its salt, all of the polar peptoid monomers can be the same, they can all be different, or a combination thereof.

[0159] In some embodiments, the peptoid polymer or its salt further comprises substituents X and Y such that the peptoid polymer has the sequence X-[H a P b ] n -Y、X-[P b H a ] n -Y、X-[H a Pb H c P d ] n -Y, or X-[P b H a P d H c ] n In some embodiments, X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, acetyl, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 Carboxyalkyl, and halogen. Optionally, X and Y form a covalent bond together. The formation of the covalent bond between X and Y results in a cyclized form of the peptoid polymer.

[0160] In some embodiments, X and Y are H, optionally substituted C 1-8 Alkylamino, -OH, -SH, acetyl, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 In other embodiments, X or Y is a secondary or tertiary amine.

[0161] In some embodiments, the peptoid polymer or its salt further includes a sequence Z, which includes one or more hydrophobic peptoid monomers and / or one or more polar peptoid monomers. Z may be located before the first subunit, after the last subunit, and / or between one or more subunits. In some instances, Z includes one or more hydrophobic peptoid monomers. In other instances, Z includes one or more polar peptoid monomers. In a specific instance, Z includes one or more hydrophobic peptoid monomers and one or more polar peptoid monomers. Z may include a plurality of continuous hydrophobic peptoid monomers, followed by a plurality of continuous polar peptoid monomers, or vice versa. Alternatively, Z may include a plurality of continuous hydrophobic peptoid monomers, followed by a plurality of continuous polar peptoid monomers, followed by additional hydrophobic peptoid monomers, and so on. When there is more than one hydrophobic peptoid monomer in sequence Z, all hydrophobic peptoid monomers may be of the same type, they may be different from each other, or a combination thereof. Similarly, when there is more than one polar peptoid monomer in sequence Z, all polar peptoid monomers may be of the same type, they may be different from each other, or a combination thereof. In some embodiments, the peptoid polymers of the present invention or their salts include more than one instance of sequence Z (e.g., 2, 3, 4, 5, or more). In such cases, all instances of Z may be the same, they may be different from each other, or a combination thereof. In a particular embodiment, sequence Z includes 1, 2, 3, 4, or more hydrophobic peptoid monomers. In yet other embodiments, sequence Z includes 1, 2, 3, 4, or more polar peptoid monomers.

[0162] In some embodiments, n is between 2 and 50 (e.g., n is 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, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50). In some examples, n is between 2 and 10 (e.g., n is 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, n is between 3 and 25. In some embodiments, n is between 5 and 25. In some embodiments, n is between 8 and 50. In some embodiments, n is between 8 and 25. In some embodiments, n is between 8 and 20. In some embodiments, n can be from about 10 to about 28, from about 12 to about 26, from about 14 to about 24, from about 16 to about 22, or from about 18 to about 20. In some embodiments, n can be from about 8 to about 50, from about 8 to about 45, from about 8 to about 40, from about 8 to about 35, from about 8 to about 30, from about 10 to about 25, from about 10 to about 20, or from about 10 to about 15.

[0163] In another aspect, the present invention provides a peptoid polymer or a salt thereof, comprising: (a) a subunit comprising two first hydrophobic peptoid monomers H and two first polar peptoid monomers P, and (b) two second hydrophobic peptoid monomers located at the C-terminus of the peptoid polymer, arranged such that the peptoid polymer has a sequence [H2P2] n H2 or [P2H2] n H2, wherein the subscript n representing the number of subunits in the peptoid polymer is between 1 and 50.

[0164] In still another aspect, the present invention provides a peptoid polymer or a salt thereof, comprising: (a) a subunit comprising two first hydrophobic peptoid monomers H and two first polar peptoid monomers P, and (b) two second polar peptoid monomers located at the C-terminus of the peptoid polymer, arranged such that the peptoid polymer has a sequence [H2P2] n P2 or [P2H2] n P2, wherein the subscript n representing the number of subunits in the peptoid polymer is between 1 and 50.

[0165] In various embodiments, all of the hydrophobic peptoid monomers can be the same, they can all be different, or a combination thereof. Similarly, all of the polar peptoid monomers can be the same, they can all be different, or a combination thereof.

[0166] In some embodiments, the peptoid polymer or its salt further comprises substituents X and Y such that the peptoid polymer has the sequence X-[H2P2] n H2-Y, X-[P2H2] n H2-Y, X-[H2P2] n P2-Y, or X-[P2H2] n In some embodiments, X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, acetyl, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 Carboxyalkyl, and halogen. Optionally, X and Y form a covalent bond together. The formation of the covalent bond between X and Y results in a cyclized form of the peptoid polymer.

[0167] In some embodiments, X and Y are H, optionally substituted C 1-8 Alkylamino, -OH, -SH, acetyl, carboxyl, optionally substituted C1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 In other embodiments, X or Y is a secondary or tertiary amine.

[0168] In some embodiments, n is between 1 and 50 (e.g., n is 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, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50). In some examples, n is between 1 and 10 (e.g., n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, n is between 1 and 25. In some embodiments, n is between 3 and 25. In some embodiments, n is between 5 and 25. In some embodiments, n is between 8 and 50. In some embodiments, n is between 8 and 25. In some embodiments, n is between 8 and 20. In some embodiments, n can be about 10 to about 28, about 12 to about 26, about 14 to about 24, about 16 to about 22, or about 18 to about 20. In some embodiments, n can be about 8 to about 50, about 8 to about 45, about 8 to about 40, about 8 to about 35, about 8 to about 30, about 10 to about 25, about 10 to about 20, or about 10 to about 15.

[0169] In some embodiments, the peptoid polymer or its salt includes Compound 62, Compound 63, Compound 67, Compound 73, Compound 76, Compound 86, or Compound 87 (i.e., when n is 2). In some examples, the peptoid polymer includes Compound 76. In other embodiments, the peptoid polymer or its salt includes Compound 81 (i.e., when n is 1).

[0170] In some embodiments, the first and / or second hydrophobic peptoid monomers are independently selected from the group consisting of:

[0171]

[0172]

[0173] wherein the subscript m is the number of repeating units and is between 1 and 10 (e.g., m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, the repeating units, m, can be between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, or 1 and 10.

[0174] In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more hydrophobic peptoid monomers in the peptoid polymer or its salt have a side chain (e.g., R 1 ), the side chains include independently selected alkyl groups wherein the alkyl group has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 or more carbon atoms. In a specific embodiment, the alkyl group has 5 carbon atoms. In some examples, the 5-carbon alkyl group is a pentyl group. In other examples, the 5-carbon alkyl group is a substituted butyl group (e.g., 1-methylbutyl, 2-methylbutyl, and 3-methylbutyl, etc.). In yet other examples, the 5-carbon alkyl group is a substituted propyl group (e.g., 1-ethylpropyl, and 1,2-dimethylpropyl, etc.).

[0175] In some embodiments, the first and / or second polar peptoid monomers are independently selected from the group consisting of:

[0176]

[0177]

[0178] wherein the subscript m is the number of repeating units and is between 1 and 10 (i.e., m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some embodiments, the repeating units, m, can be between 1 and 2, 1 and 3, 1 and 4, 1 and 5, 1 and 6, 1 and 7, 1 and 8, 1 and 9, or 1 and 10.

[0179] In some embodiments, the first and / or second polar peptoid monomers have side chains containing hydroxyl groups (e.g., R 1 In other embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more polar peptoid monomers in the peptoid polymer or its salt have independently selected C 1-18 Hydroxyalkyl (e.g., independently selected C 1-6 hydroxyalkyl) side chain (e.g., R 1 ). In some examples, each C 1-18Hydroxyalkyl is C 1-6 In a specific example, each C 1-6 Hydroxylalkyl is the same C 1-6 In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more polar peptoid monomers in the peptoid polymer or its salt have side chains (e.g., R 1 ), wherein the length of the alkyl group in the hydroxyalkyl group is selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or more carbon atoms. In certain embodiments, the hydroxyalkyl group contains 1, 2, 3, 4, 5, 6, 7, or 8 hydroxy substituents.

[0180] In some embodiments, none of the polar peptoid monomers include an optionally substituted C 1-18 The side chain of a hydroxyalkyl group (e.g., R 1 ).

[0181] In some embodiments, the side chain of the polar peptoid monomer (e.g., R 1 ) includes (4- to 10-membered heterocycloalkyl) (C 1-6 (alkylene) or (5- to 10-membered heteroaryl) (C 1-6 In some embodiments, the alkylene moiety is a methylene moiety. In some embodiments, the alkylene moiety is a n-propylene moiety. In some embodiments, at least one member of the 4-, 5-, 6-, 7-, 8-, 9-, or 10-membered ring is O. In other embodiments, at least one member of the 4-, 5-, 6-, 7-, 8-, 9-, or 10-membered ring is N.

[0182] The heterocycloalkyl moiety can be, but is not limited to, 4- to 8-membered rings, 4- to 6-membered rings, or 5- to 6-membered rings. The heterocycloalkyl moiety can be, for example, azetidinyl, pyrrolidinyl, piperidinyl, aziridine, azanyl, pyrazolidinyl, imidazolidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, oxirane, tetrahydrothienyl, tetrahydrothiopyranyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, or morpholinyl. In some embodiments, the heterocycloalkyl moiety is selected from pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl, tetrahydrofuran-2-yl, and tetrahydrofuran-3-yl. In some embodiments, the heterocycloalkyl moiety is pyrrolidin-1-yl. In some embodiments, the heterocycloalkyl moiety is tetrahydrofuran-2-yl. In some embodiments, one or more carbocyclic ring members in pyrrolidinyl or tetrahydrofuranyl are substituted by oxygen.

[0183] In some embodiments, the peptoid polymer comprises and / or In some examples, the peptoid polymer includes Compound 63, Compound 76, Compound 86, and / or Compound 87. In some embodiments, all polar peptoid monomers are In other embodiments, all polar peptoid monomers are In some examples, the peptoid polymer is Compound 76, Compound 86, or Compound 87.

[0184] The heteroaryl moiety can be, but is not limited to, a 5- to 10-membered ring, a 5- to 9-membered ring, or a 5- to 6-membered ring. The heteroaryl moiety can be, for example, an indolyl, quinolyl, quinoxalinyl, quinazolinyl, phthalazinyl, cinnolinyl, benzofuranyl, pyrrolyl, pyridyl, imidazolyl, pyrazolyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl, or furanyl. In some embodiments, the heteroaryl moiety is selected from furan-2-yl, furan-3-yl, thien-2-yl, thien-3-yl, pyrrol-1-yl, pyrrol-2-yl, and pyrrol-3-yl.

[0185] In some embodiments, the side chain (e.g., R 1 ) includes (2-oxopyrrolidin-1-yl) (C 1-4 In some embodiments, the side chain (e.g., R 1 ) includes (tetrahydrofuran-2-yl)(C 1-4 In some embodiments, the side chain (e.g., R 1 ) includes (furan-2-yl)(C 1-4 alkylene).

[0186] In some embodiments, the peptoid polymer comprises In certain embodiments, all polar peptoid monomers are In some instances, the peptoid polymer is Compound 73.

[0187] In some embodiments, the side chain of the polar peptoid monomer (e.g., R 1 ) include 2- to 20-membered alkoxy groups. For example, the side chain (e.g., R 1 ) may include a 2-12 membered alkoxy group having 1-4 oxygen atoms, or a 2-6 membered alkoxy group having 1 or 2 oxygen atoms. 1 ) includes –CH2CH2OR′, where R′ is C 1-6 In some embodiments, the side chain (e.g., R 1 ) including –CH2CH2O(CH2CH2O) n R′, where R′ is C 1-6 The subscript n is 1, 2, or 3.

[0188] In some embodiments, the side chain (e.g., R 1 ) includes (C 1-6 Alkoxy) (C 1-6 In some embodiments, the side chain (e.g., R 1 ) includes (oligo[ethylene glycol]) groups or (oligo[propylene glycol]) groups. In some embodiments, the side chain (e.g., R 1 ) includes methoxyethyl. In some embodiments, the peptoid polymer includes In certain embodiments, all polar peptoid monomers are In some instances, the peptoid polymer is Compound 62.

[0189] In some embodiments, the oligo(ethylene glycol) moiety is a 2-(2-(2-methoxyethoxy)ethoxy)ethyl moiety. In some embodiments, the peptoid polymer comprises In certain embodiments, all polar peptoid monomers are In some instances, the peptoid polymer is compound 67.

[0190] In yet another aspect, the present invention provides a peptoid polymer or a salt thereof, which includes one or more hydrophobic peptoid monomers and one or more polar peptoid monomers. In some embodiments, none of the polar peptoid monomers include an optionally substituted C 1-18 The side chain of a hydroxyalkyl group (e.g., R 1 In some embodiments, the side chain of the polar peptoid monomer (e.g., R 1 ) includes (4- to 10-membered heterocycloalkyl) (C 1-6 In other embodiments, the side chain of the polar peptoid monomer (e.g., R 1) includes (5- to 10-membered heteroaryl) (C 1-6 In some embodiments, the side chain of the polar peptoid monomer (e.g., R 1 ) include 2- to 20-membered alkoxy groups. For example, the side chain (e.g., R 1 ) may include a 2-12 membered alkoxy group having 1-4 oxygen atoms, or a 2-6 membered alkoxy group having 1 or 2 oxygen atoms. 1 ) includes –CH2CH2OR′, where R′ is C 1-6 In some embodiments, the side chain (e.g., R 1 ) including –CH2CH2O(CH2CH2O) n R′, where R′ is C 1-6 alkyl and the subscript n is 1, 2, or 3. In some embodiments, the side chain of the polar peptoid monomer (e.g., R 1 ) includes (C 1-6 Alkoxy) (C 1-6 In some embodiments, the side chain of the polar peptoid monomer (e.g., R 1 ) includes an (oligo[ethylene glycol]) group or an (oligo[propylene glycol]) group.

[0191] For example, the alkylene moiety can be a straight chain alkylene moiety, such as methylene, ethylene, n-propylene (i.e., -CH2CH2CH2-), or n-butylene (i.e., -CH2CH2CH2CH2-). The alkylene linker can also be branched, such as in the case of secondary butylene (i.e., -CH(CH3)CH2CH2-) or isobutylene (i.e., -CH2CH(CH3)CH2-). In some embodiments, the alkylene moiety is methylene (i.e., -CH2-). In some embodiments, the alkylene moiety is n-propylene. In some embodiments, at least one member of the 4-, 5-, 6-, 7-, 8-, 9-, or 10-membered ring is O. In other embodiments, at least one member of the 4-, 5-, 6-, 7-, 8-, 9-, or 10-membered ring is N.

[0192] The heterocycloalkyl moiety can be, but is not limited to, 4- to 8-membered rings, 4- to 6-membered rings, or 5- to 6-membered rings. The heterocycloalkyl moiety can be, for example, azetidinyl, pyrrolidinyl, piperidinyl, aziridine, azanyl, pyrazolidinyl, imidazolidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, oxirane, tetrahydrothienyl, tetrahydrothiopyranyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, or morpholinyl. In some embodiments, the heterocycloalkyl moiety is selected from pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl, tetrahydrofuran-2-yl, and tetrahydrofuran-3-yl. In some embodiments, the heterocycloalkyl moiety is pyrrolidin-1-yl. In some embodiments, the heterocycloalkyl moiety is tetrahydrofuran-2-yl. In some embodiments, one or more carbocyclic ring members in pyrrolidinyl or tetrahydrofuranyl are substituted by oxygen.

[0193] In some embodiments, the peptoid polymer comprises and / or In some examples, the peptoid polymer includes Compound 63, Compound 76, Compound 86, and / or Compound 87. In some embodiments, all polar peptoid monomers are In other embodiments, all polar peptoid monomers are In some examples, the peptoid polymer is Compound 76, Compound 86, or Compound 87.

[0194] The heteroaryl moiety can be, but is not limited to, a 5- to 10-membered ring, a 5- to 9-membered ring, or a 5- to 6-membered ring. The heteroaryl moiety can be, for example, an indolyl, quinolyl, quinoxalinyl, quinazolinyl, phthalazinyl, cinnolinyl, benzofuranyl, pyrrolyl, pyridyl, imidazolyl, pyrazolyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl, or furanyl. In some embodiments, the heteroaryl moiety is selected from furan-2-yl, furan-3-yl, thien-2-yl, thien-3-yl, pyrrol-1-yl, pyrrol-2-yl, and pyrrol-3-yl.

[0195] In some embodiments, the side chain (e.g., R 1 ) includes (2-oxopyrrolidin-1-yl) (C 1-4 In some embodiments, the side chain (e.g., R 1 ) includes (tetrahydrofuran-2-yl)(C 1-4 In some embodiments, the side chain (e.g., R 1 ) includes (furan-2-yl)(C 1-4 alkylene).

[0196] In some embodiments, the peptoid polymer comprises In certain embodiments, all polar peptoid monomers are In some instances, the peptoid polymer is Compound 73.

[0197] In some embodiments, the side chain (e.g., R 1 ) includes methoxyethyl. In some embodiments, the peptoid polymer includes In certain embodiments, all polar peptoid monomers are In some instances, the peptoid polymer is Compound 62.

[0198] In some embodiments, the oligo(ethylene glycol) moiety is a 2-(2-(2-methoxyethoxy)ethoxy)ethyl moiety. In some embodiments, the peptoid polymer comprises In certain embodiments, all polar peptoid monomers are In some instances, the peptoid polymer is compound 67.

[0199] In some embodiments, the one or more hydrophobic peptoid monomers are each independently selected from the group consisting of: wherein the subscript m is the number of repeating units and is between 1 and 10.

[0200] In some embodiments, the one or more polar peptoid monomers are each independently selected from the group consisting of:

[0201] In some embodiments, the peptoid polymer further comprises substituents X and Y located at the N-terminus and C-terminus of the peptoid polymer, respectively. In some embodiments, X and Y are independently selected from H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, acetyl, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 A group consisting of carboxyl, alkyl, and halogen. Optionally, X and Y form a covalent bond together. The formation of the covalent bond between X and Y results in a cyclized form of the peptoid polymer.

[0202] In some embodiments, X and Y are H, optionally substituted C 1-8 Alkylamino, -OH, -SH, acetyl, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8Alkylthio, optionally substituted C 1-8 In other embodiments, X or Y is a secondary or tertiary amine.

[0203] In some embodiments, the sequence length of the peptoid polymers described herein is between 6 and 50. In some embodiments, the sequence length of the peptoid polymers is between 10 and 50. In some embodiments, the sequence length of the peptoid polymers is between 16 and 100. In some embodiments, the sequence length of the peptoid polymers is between 16 and 50. In some embodiments, the sequence length of the peptoid polymers is between 16 and 40. In some embodiments, the sequence length of the peptoid polymers can be about 20 to about 56, about 24 to about 52, about 28 to about 48, about 32 to about 44, or about 36 to about 40. In some embodiments, the sequence length of the peptoid polymers can be about 16 to about 100, about 16 to about 90, about 16 to about 80, about 16 to about 70, about 16 to about 60, about 20 to about 50, about 20 to about 40, or about 20 to about 30. In some embodiments, the sequence length of the peptoid polymer can be 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, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 , 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, or more.

[0204] In some embodiments, between about 1% and about 99% (e.g., about 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, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 94, 95, 96, 97, 98, or 99%) of the peptoid monomers are hydrophobic peptoid monomers. In other embodiments, between about 1% and about 99% (e.g., about 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, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65 94, 95, 96, 97, 98, or 99%) of the peptoid monomers are polar peptoid monomers.

[0205] In some embodiments, about 5% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 95% of the peptoid monomers are polar.

[0206] In some embodiments, about 10% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 90% of the peptoid monomers are polar.

[0207] In some embodiments, about 15% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 85% of the peptoid monomers are polar.

[0208] In some embodiments, about 20% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 80% of the peptoid monomers are polar.

[0209] In some embodiments, about 25% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 75% of the peptoid monomers are polar.

[0210] In some embodiments, about 30% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 70% of the peptoid monomers are polar.

[0211] In some embodiments, about 35% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 65% of the peptoid monomers are polar.

[0212] In some embodiments, about 40% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 60% of the peptoid monomers are polar.

[0213] In some embodiments, about 45% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 55% of the peptoid monomers are polar.

[0214] In some embodiments, about 50% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 50% of the peptoid monomers are polar.

[0215] In some embodiments, about 55% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 45% of the peptoid monomers are polar.

[0216] In some embodiments, about 60% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 40% of the peptoid monomers are polar.

[0217] In some embodiments, about 65% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 35% of the peptoid monomers are polar.

[0218] In some embodiments, about 70% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 30% of the peptoid monomers are polar.

[0219] In some embodiments, about 75% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 25% of the peptoid monomers are polar.

[0220] In some embodiments, about 80% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 20% of the peptoid monomers are polar.

[0221] In some embodiments, about 85% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 15% of the peptoid monomers are polar.

[0222] In some embodiments, about 90% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 10% of the peptoid monomers are polar.

[0223] In some embodiments, about 95% of the peptoid monomers in the peptoid polymer or salt thereof are hydrophobic and about 5% of the peptoid monomers are polar.

[0224] In certain embodiments, the peptoid polymer or its salt comprises about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95% hydrophobic peptoid monomers by mass. In other embodiments, the peptoid polymer or its salt comprises about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95% polar peptoid monomers by mass.

[0225] In some embodiments, the peptoid polymer forms a helical structure. In some embodiments, the helical structure adopts a structure similar to a polyproline helix. In certain instances, the peptoid polymer forms a polyproline I helix. In certain other instances, the peptoid polymer forms a polyproline II helix. In some embodiments, the helical structure is adopted when the peptoid polymer contains at least one N-aryl side chain. In some embodiments, the N-aryl side chain is a Nep monomer.

[0226] In some embodiments, the peptoid polymer or its salt reduces or inhibits ice crystal formation at a temperature within about 0°C to about -20°C (e.g., at about 0, -1, -2, -3, -4, -5, -6, -7, -8, -9, -10, -11, -12, -13, -14, -15, -16, -17, -18, -19, or -20°C). In other embodiments, the peptoid polymer or its salt reduces or inhibits ice crystal formation at a temperature within about -20°C to about -40°C (e.g., at about -20, -21, -22, -23, -24, -25, -26, -27, -28, -29, -30, -31, -32, -33, -34, -35, -36, -37, -38, -39, or -40°C). In certain embodiments, the peptoid polymer or its salt reduces or inhibits ice crystal formation at about -20°C. In certain other embodiments, the peptoid polymer or its salt reduces or inhibits ice crystal formation at a temperature within about -40°C to about -200°C (e.g., at about -40, -45, -50, -55, -60, -65, -70, -75, -80, -85, -90, -95, -100, -105, -110, -115, -120, -125, -130, -135, -140, -145, -150, -155, -160, -165, -170, -175, -180, -185, -190, -195, or -200°C). In some examples, the peptoid polymer or its salt reduces or inhibits ice crystal formation at about -196°C.

[0227] In some embodiments, the peptoid polymer or a salt thereof reduces or inhibits ice crystal formation at a temperature within about 0°C to about -200°C, within about -10°C to about -190°C, within about -20°C to about -180°C, within about -30°C to about -170°C, within about -40°C to about -160°C, within about -50°C to about -150°C, within about -60°C to about -140°C, within about -70°C to about -140°C, within about -80°C to about -130°C, within about -90°C to about -120°C, or within about -100°C to about -110°C.

[0228] In some embodiments, the concentration of the peptoid polymer and / or its salt (e.g., present in a composition, formulation, or product such as a cryoprotectant, antifreeze, frozen food, or beauty care product) is between about 100 nM and about 1,000 mM. In certain embodiments, the concentration of the peptoid polymer and / or its salt (e.g., present in a composition, formulation, or product such as a cryoprotectant, antifreeze, frozen food, or beauty care product) is between about 100 nM and about 250 nM, between about 250 nM and about 500 nM, between about 500 nM and about 750 nM, between about 750 nM and about 1 μM, between about 1 μM and about 5 μM, between about 5 μM and about 25 μM, between about 25 μM and about 50 μM, between about 50 μM and about 100 μM. In some embodiments, the present invention relates to an aqueous solution of at least about 1% to about 5% sodium bicarbonate, wherein the amount of the aqueous solution of at least about 1% sodium bicarbonate is about 0.1% to about 0.1% sodium bicarbonate, wherein the amount of the aqueous solution of at least about 1% sodium bicarbonate is about 0.1% to about 0.1% sodium bicarbonate, and wherein the amount of the aqueous solution of at least about 1% sodium bicarbonate is about 0.1% to about 0.1% sodium bicarbonate. In other embodiments, the concentration of the peptoid polymer and / or its salt (e.g., present in a composition, formulation, or product such as a cryoprotectant, antifreeze, frozen food, or beauty care product) is about 100 nM, about 1 μM, about 10 μM, about 100 μM, about 1 mM, about 10 mM, about 100 mM, or about 1,000 mM. In specific embodiments, the concentration of the peptoid polymer and / or its salt is about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mM.

[0229] For embodiments of the present invention in which the peptoid polymer is prepared or used as a salt, any number of salt forms may be prepared or used. Non-limiting examples of suitable salt forms include hydrochlorides, acetates, sulfates, phosphates, maleates, citrates, mesylates, nitrates, tartrates, and gluconates. In addition, any suitable combination of salt forms of the peptoid polymer may be prepared or used.

[0230] In some embodiments, the peptoid polymer or its salt comprises a peptoid polymer selected from the group consisting of: Compound 59, Compound 60, Compound 61, Compound 62, Compound 63, Compound 64, Compound 65, Compound 66, Compound 67, Compound 68, Compound 69, Compound 70, Compound 71, Compound 72, Compound 73, Compound 74, Compound 75, Compound 76, Compound 77, Compound 78, Compound 79, Compound 80, Compound 81, Compound 82, Compound 83, Compound 84, Compound 85, Compound 86, and Compound 87. In some embodiments, the peptoid polymer or its salt comprises a peptoid polymer selected from the group consisting of: Compound 88, Compound 89, Compound 90, Compound 91, Compound 92, Compound 93, Compound 94, Compound 95, Compound 96, Compound 97, Compound 98, Compound 99, Compound 100, and Compound 101. In specific embodiments, the peptoid polymer is Compound 62, 63, 67, 73, 76, 86, or 87. In some examples, the peptoid polymer is compound 76, 86, or 87.

[0231] In some embodiments, the peptoid polymer is not a peptoid polymer listed in Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, or Table 9.

[0232] B. Peptoid-Peptide Hybrids

[0233] On the other hand, the present invention provides peptoid-peptide hybrids or their salts. In some embodiments, peptoid-peptide hybrids or their salts include peptoid polymers as described herein and one or more amino acids. Amino acids can be naturally occurring amino acids or variants thereof. In some embodiments, peptoid-peptide hybrids or their salts include about 1 and 10 amino acids (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids). In other embodiments, peptoid-peptide hybrids or their salts include about 10 and 100 amino acids (e.g., about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 amino acids). In some embodiments, peptoid-peptide hybrids or their salts include greater than about 100 amino acids. In other embodiments, the peptoid-peptide hybrid or salt thereof includes between about 2 and 50 peptoid monomers (e.g., 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, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 42, 44, 45, 46, 47, 48, 49, or 50 peptoid monomers) and at least about between 1 and 100 amino acids (e.g., at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 amino acids).

[0234] In some embodiments, the amino acid is located at the N- and / or C-terminal of the peptoid polymer. In other embodiments, the amino acid is located between one or more subunits of the peptoid polymer. In a specific embodiment, the amino acid is located at one or both ends of the peptoid polymer and between one or more subunits. In the example where the peptoid-peptide hybrid or its salt includes more than two amino acids, the amino acids can all be continuous, or only a part of them can be continuous. Alternatively, all amino acids can be separated by one or more subunits.

[0235] In some embodiments, the amino acid is a D-amino acid. In other embodiments, the amino acid is an L-amino acid. In some other embodiments, the peptoid-peptide hybrid or its salt includes a combination of D- and L-amino acids. In some embodiments, one or more amino acids are selected from the group consisting of: alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, arginine, lysine, leucine, methionine, asparagine, proline, glutamine, serine, threonine, valine, tryptophan, tyrosine, and combinations thereof. In some instances, one or more amino acids are selected from the group consisting of isoleucine, leucine, serine, threonine, alanine, valine, arginine, and combinations thereof.

[0236] In certain embodiments, the peptoid-peptide hybrid or its salt comprises about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95% hydrophobic peptoid monomers by mass. In other embodiments, the peptoid-peptide hybrid or its salt comprises about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95% polar peptoid monomers by mass.

[0237] For embodiments of the present invention in which the peptoid-peptide hybrid is prepared or used as a salt, any number of salt forms may be prepared or used. Non-limiting examples of suitable salt forms include hydrochlorides, acetates, sulfates, phosphates, maleates, citrates, mesylates, nitrates, tartrates, and gluconates. In addition, any suitable combination of salt forms of peptoid polymers may be prepared or used.

[0238] In some embodiments, the peptoid-peptide hybrid is not a peptoid-peptide hybrid described in SEQ ID NOS: 3-5 or Table 10. In SEQ ID NOS: 3, 4, and 5, Xaa represents any amino acid.

[0239] C. Synthesis Method

[0240] On the other hand, the present invention provides a method for synthesizing a peptoid polymer and / or its salt or a peptoid-peptide hybrid and / or its salt. The peptoid polymer, peptoid polymer salt, peptoid-peptide hybrid and peptoid-peptide hybrid salt of the present invention can be prepared from readily available raw materials using the general methods and processes described herein. It should be understood that, unless otherwise stated, other process conditions may also be used when typical or preferred process conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.) are given. Optimal reaction conditions may vary with the specific reactants or solvents used, but such conditions may be determined by a skilled person in the art through conventional optimization processes.

[0241] The peptoid polymers, peptide polymer salts, peptoid-peptide hybrids, and peptoid-peptide hybrid salts of the present invention can be prepared by those skilled in the art of organic synthesis from known or commercially available raw materials and reagents. Solvents and reagents were purchased from commercial channels and used without further purification.

[0242] In some embodiments, the sub-monomer method ( Figure 1 ) is used for the synthesis of peptoids, where each N-substituted glycine monomer is assembled from two readily available "submonomers". Similar to peptide synthesis, the synthesis of oligomeric peptoids is based on the robust chemistry of standard solid phase methods. Each cycle of monomer addition consists of two steps: an acylation step and a nucleophilic substitution step. In some embodiments, solid phase assembly does not require N-protected monomers because there are no reactive side chain functional groups that need to be protected. Those skilled in the art will recognize that there are many solid phase synthesis methods, including automated robotic synthesizers. In some embodiments, the synthesizer used is manufactured by Protein Technologies, Inc. X Multiplex Peptide Synthesizer. In some embodiments, the synthesizer used is an Overture Peptide Synthesizer manufactured by Protein Technologies, Inc. In other embodiments, peptoids are artificially synthesized using conventional organic chemistry methods known in the art. The same techniques or techniques similar to those described above can be applied to the synthesis of peptoid-peptide oligomers by providing appropriate amino acids at appropriate times during the synthesis in place of the peptoid monomers.

[0243] As a non-limiting example, a peptoid polymer can be synthesized on 100 mg of Rink amide resin (NovaBiochem; 0.49 mmol / g). Rink amide resin (100 mg) can be washed twice in 1.5 mL of DCM and then swelled in 1.5 mL of DMF. The swelling step can be performed twice. The Fmoc protecting group can be removed from the resin by adding 20% ​​piperidine / DMF. The mixture is stirred for 10 minutes, drained, and the piperidine treatment is repeated, followed by thorough washing with DMF (1.5 mL, 5 times). The first monomer can be added manually by reacting 37 mg of bromoacetic acid (0.27 mmol; Sigma-Aldrich) and 189 μL of DIEA (1.08 mmol; Chem Impex International) in 2 mL of DCM on a shaking platform for 30 minutes at room temperature, followed by thorough washing with DCM (2 mL, 5 times) and DMF (2 mL, 5 times). Bromoacylated resin can be incubated with 2mL of 1M amine sub-monomer in DMF on a shaking platform at room temperature for 30 minutes, and then fully washed with DMF (2mL, 5 times). After the initial manual loading of bromoacetic acid, the first sub-monomer substitution step and all subsequent bromoacetylation and amine substitution steps can be carried out by a robot synthesizer until the desired oligomer length is obtained. Automatic bromoacetylation step can be carried out by adding 1,660μL of 1.2M bromoacetic acid in DMF and 400μL of DIC (Chem Impex International). The mixture is stirred for 20 minutes, drained, and washed with DMF (2mL, 3 times). Then, the 1M solution of the sub-monomer (2mmol) of 2mL in DMF can be added to introduce side chains by nucleophilic substitution of bromide. The mixture can be stirred for 20 minutes, drained, washed with DMF (2mL, 3 times) and washed with DCM (2mL, 3 times). The peptoid-resin can be cleaved in 20% HFIP (Alfa Aesar) (v / v) in 2 mL DCM at room temperature. The cleavage can be performed in a glass tube under constant stirring for 30 minutes. The HFIP / DCM can be evaporated under a stream of nitrogen. The final product can be dissolved in 50% ACN in 5 mL HPLC grade H2O and filtered with a 0.5 pm stainless steel sintered syringe tip filter (Upchurch Scientific). The Beckman Coulter System Gold instrument can be used at room temperature at C 18 Reversed phase analytical HPLC column (Peeke Scientific, 5 pm, Peptoid oligomers were analyzed on a 40×50 mm (2.0×50 mm) plate. A linear gradient of 5-95% acetonitrile / water (0.1% TFA, Acros Organics) over 20 minutes was used with a flow rate of 0.7 mL / min. To remove any traces of HFIP in the sample solution, the linear precursor dissolved in 50% ACN / H2O was freeze-dried overnight.

[0244] Peptoid polymers and peptoid-peptide hybrids can be analyzed by electrospray ionization (ESI) mass spectrometry. Typically, 0.5-2mL of 1-5 μM peptoid polymers or peptoid-peptide hybrids to be analyzed are prepared in 50% deionized water / 50% HPLC grade ACN with 1% organic acids such as trifluoroacetic acid. The prepared sample is ionized by electron bombardment, so that the molecules are split into charged fragments. The fragments are then accelerated and exposed to an electric or magnetic field, and separated according to the mass-to-charge ratio of the ions. Ions are detected by mechanisms such as electron multipliers that can detect charged particles. Peptoids and peptoid-peptide hybrids are identified by associating mass with the mass of identification or by characteristic fragmentation patterns.

[0245] D. How to use

[0246] In some aspects, the present invention provides a cryoprotectant. In some embodiments, the cryoprotectant comprises a peptoid polymer and / or a salt thereof as described herein, a peptoid-peptide hybrid as described herein and / or a salt thereof, or a combination thereof. In other embodiments, the cryoprotectant further comprises a compound selected from the group consisting of: an ionic substance, a permeating cryoprotectant, a non-permeating cryoprotectant, an antioxidant, a cell membrane stabilizing compound, a water channel protein or other channel forming compound, an alcohol, a sugar, a sugar derivative, a non-ionic surfactant, a protein, dimethyl sulfoxide (DMSO), polyethylene glycol (PEG), polypropylene glycol (PPG), Polyvinyl pyrrolidone, polyvinyl alcohol, hyaluronic acid, formamide, natural or synthetic hydrogels, and combinations thereof. In specific embodiments, permeable cryoprotectants permeate the cell membrane and reduce the concentration of intracellular water, thereby reducing the amount of ice formed at any temperature. In other specific embodiments, non-permeable cryoprotectants induce changes in colloid osmotic pressure and alter the association of cell membranes with extracellular water through induced ionic interactions.

[0247] In some examples, the cryoprotectant further includes an alcohol selected from the group consisting of propylene glycol, ethylene glycol, glycerol, methanol, butylene glycol, adonitol, ethanol, trimethylene alcohol, diethylene glycol, polyethylene oxide, erythritol, sorbitol, xylitol, polypropylene glycol, 2-methyl-2,4-pentanediol (MPD), mannitol, inositol, dithiothitol, 1,2-propylene glycol, and combinations thereof.

[0248] In other examples, the cryoprotectant further includes a sugar selected from the group consisting of monosaccharides, disaccharides, polysaccharides, and combinations thereof. In a specific example, the sugar is selected from the group consisting of glucose, 3-O-methyl-D-pyranose glucose, galactose, arabinose, fructose, xylose, mannose, sucrose, trehalose, lactose, maltose, raffinose, dextran, and combinations thereof.

[0249] In other examples, the cryoprotectant further comprises PEG or a plurality of different PEG compounds. In some examples, at least one PEG compound has a molecular weight of less than about 3,000 g / mol (e.g., less than about 3,000, 2,950, 2,900, 2,850, 2,800, 2,750, 2,700, 2,650, 2,600, 2,550, 2,500, 2,450, 2,400, 2,350, 2,300, 2,250, 2,200, 2,150, 2,100, 2,050, 2,000, 1,950, 1,900, 1,850, 1, 700, 650, 600, 550, 500, 450, 400, 350, 300, 250, 200, 150, or 100 g / mol). In certain examples, at least one PEG compound has an average molecular weight between about 200 g / mol and 400 g / mol (e.g., about 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, or 400 g / mol). In some examples, the cryoprotectant fluid includes PEG or a plurality of PEG compounds selected from the group consisting of PEG 200, PEG 300, PEG 400, and combinations thereof.

[0250] In other examples, the cryoprotectant further comprises PPG or a plurality of different PPG compounds. In some examples, at least one PPG compound has a molecular weight of less than about 3,000 g / mol (e.g., less than about 3,000, 2,950, 2,900, 2,850, 2,800, 2,750, 2,700, 2,650, 2,600, 2,550, 2,500, 2,450, 2,400, 2,350, 2,300, 2,250, 2,200, 2,150, 2,100, 2,050, 2,000, 1,950, 1,900, 1,850, 1, 700, 650, 600, 550, 500, 450, 400, 350, 300, 250, 200, 150, or 100 g / mol). In certain examples, at least one PPG compound has an average molecular weight between about 200 and 400 g / mol (e.g., about 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, or 400 g / mol).

[0251] In other examples, the cryoprotectant further comprises a protein selected from the group consisting of ovalbumin, bovine serum albumin, human serum albumin, gelatin, and combinations thereof. In still other examples, the cryoprotectant further comprises a natural or synthetic hydrogel, wherein the natural or synthetic hydrogel comprises chitosan, hyaluronic acid, or a combination thereof. In yet other examples, the cryoprotectant further comprises a nonionic surfactant selected from the group consisting of polyoxyethylene lauryl ether, polysorbate 80, and combinations thereof.

[0252] Non-limiting examples of various properties of cryoprotectant solutions, such as effective concentration, viscosity, water solubility, and / or membrane permeability, can be evaluated using model cells or tissues including, but not limited to, stem cells, liver tissue or hepatocytes, kidney, intestine, heart, pancreas, genitourinary cells (e.g., sperm cells, oocytes, corpus cavernosum cells (e.g., corpus cavernosum smooth muscle cells, corpus cavernosum epithelial cells), bladder cells, urethral cells, ureteral cells, kidney cells, testicular cells), bone marrow, primary cells, organoids, and other biological tissues for cryopreservation.

[0253] In some embodiments, the cryoprotectant reduces or inhibits ice crystal formation at a temperature within about 0°C to about -20°C (e.g., at about 0, -1, -2, -3, -4, -5, -6, -7, -8, -9, -10, -11, -12, -13, -14, -15, -16, -17, -18, -19, or -20°C). In other embodiments, the cryoprotectant reduces or inhibits ice crystal formation at a temperature within about -20°C to about -40°C (e.g., at about -20, -21, -22, -23, -24, -25, -26, -27, -28, -29, -30, -31, -32, -33, -34, -35, -36, -37, -38, -39, or -40°C). In certain embodiments, the cryoprotectant reduces or inhibits ice crystal formation at about -20°C. In certain other embodiments, the cryoprotectant reduces or inhibits ice crystal formation at a temperature within about -40°C to about -200°C (e.g., at about -40, -45, -50, -55, -60, -65, -70, -75, -80, -85, -90, -95, -100, -105, -110, -115, -120, -125, -130, -135, -140, -145, -150, -155, -160, -165, -170, -175, -180, -185, -190, -195, or -200°C). In some instances, the cryoprotectant reduces or inhibits ice crystal formation at about -196°C.

[0254] In some embodiments, the cryoprotectant solution reduces or inhibits ice crystal formation at temperatures within about 0°C to about -200°C, within about -10°C to about -190°C, within about -20°C to about -180°C, within about -30°C to about -170°C, within about -40°C to about -160°C, within about -50°C to about -150°C, within about -60°C to about -140°C, within about -70°C to about -140°C, within about -80°C to about -130°C, within about -90°C to about -120°C, or within about -100°C to about -110°C.

[0255] In some embodiments, the concentration of the peptoid polymer and / or its salt and / or the peptoid-peptide hybrid and / or its salt in the cryoprotectant is between about 100 nM and about 1,000 mM. In some embodiments, the concentration of the peptoid polymer and / or its salt and / or the peptoid-peptide hybrid and / or its salt in the cryoprotectant is between about 100 nM and about 250 nM, between about 250 nM and about 500 nM, between about 500 nM and about 750 nM, between about 750 nM and about 1 μM, between about 1 μM and about 5 μM, between about 5 μM and about 25 μM, between about 25 μM and about 50 μM, between about 50 μM and about 100 μM, between about 100 μM and about 25 μM. and about 250 μM, between about 250 μM and about 500 μM, between about 500 μM and about 750 μM, between about 750 μM and about 1 mM, between about 1 mM and about 10 mM, between about 10 mM and about 50 mM, between about 50 mM and about 100 mM, between about 100 mM and about 250 mM, between about 250 mM and about 500 mM, between about 500 mM and about 750 mM, or between about 750 mM and about 1,000 mM. In some embodiments, the concentration of the peptoid polymer and / or its salt and / or peptoid-peptide hybrid and / or its salt in the cryoprotectant is about 100 nM, about 1 μM, about 10 μM, about 100 μM, about 1 mM, about 10 mM, about 100 mM, or about 1,000 mM. In specific embodiments, the concentration of the peptoid polymer and / or its salt and / or peptoid-peptide hybrid and / or its salt in the cryoprotectant is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 mM.

[0256] In other aspects, the present invention provides a method for preserving a biological sample. In a specific embodiment, the biological sample has a cell composition. In some embodiments, the biological sample includes a tissue. In other embodiments, the biological sample includes an organ. In still other embodiments, the biological sample includes a cell. In a specific embodiment, the biological sample includes one or more tissues, one or more organs, one or more cells, or a combination thereof. In some embodiments, the method includes contacting the biological sample with a peptoid polymer as described herein and / or its salt, a peptoid-peptide hybrid as described herein and / or its salt, a cryoprotectant as described herein, or a combination thereof. In some examples, when a combination of a composition or a solution is used, contacting the biological sample with a composition or a solution can be completed in multiple steps. As a non-limiting example, the biological sample can be first contacted with a peptoid polymer as described herein and / or its salt, and then the biological sample can be contacted with a cryoprotectant as described herein at a later time point.

[0257] In certain examples, the tissue is a bioengineered tissue. In some examples, the biological sample is selected from the group consisting of: heart, liver, lung, kidney, pancreas, intestine, thymus, cornea, nerve cells, platelets, genitourinary cells (e.g., sperm cells, oocytes, corpus cavernosum cells (e.g., corpus cavernosum smooth muscle cells, corpus cavernosum epithelial cells), bladder cells, urethral cells, ureteral cells, kidney cells, testicular cells), embryonic cells, stem cells, bone cells, primary cells, and combinations thereof.

[0258] Cryopreservation of biological samples can be used for various purposes. Non-limiting examples include organoid preservation, primary cell preservation, stem cell (e.g., hematopoietic stem cell, embryonic stem (ES) cell, pluripotent stem cell (PSC) and induced pluripotent stem cell (iPSC)) preservation, adult cell and cell line (e.g., lymphocyte, granulocyte, immune system cell, osteocyte) preservation, embryo, sperm and oocyte preservation, tissue preservation, and organ preservation. For example, in the field of organ transplantation, the preservation of tissues, organs and other biological samples and structures is very useful. Those skilled in the art will readily understand other useful applications of the present invention to cryopreservation of biological samples.

[0259] In other aspects, the present invention provides a method for preserving one or more biomacromolecules. The biomacromolecule can be a natural or non-natural biomacromolecule. Non-limiting examples of biomacromolecules suitable for cryoprotection by the compositions and methods of the present invention include nucleic acids (e.g., DNA, RNA), amino acids, proteins, peptides, lipids, and complex structures (e.g., liposomes). In some embodiments, the method includes contacting the biomacromolecule with a peptoid polymer and / or its salt as described herein, a peptoid-peptide hybrid as described herein, and / or its salt, a cryoprotectant solution as described herein, or a combination thereof. In some instances, the biomacromolecule is an isolated protein. In a particular instance, the isolated protein is a protease protein. In some instances, when using a combination of a composition or a solution, contacting one or more biomacromolecules with a composition or a solution can be completed in multiple steps. As a non-limiting example, one or more biomacromolecules can be first contacted with a peptoid polymer and / or its salt as described herein, and then at a later time point, the biological sample is contacted with a cryoprotectant solution as described herein.

[0260] Cryoprotection of biomacromolecules using the compositions and methods of the present invention can be used for a variety of purposes. Non-limiting examples of such purposes include the preservation of DNA (e.g., genomic DNA) and RNA samples, the preservation of stem cell growth factors, and the preservation of antibodies. Those skilled in the art will readily appreciate other useful purposes and suitable applications of the compositions and methods of the present invention.

[0261] In a particular embodiment, the separated protein has been crystallized. Crystal cryoprotection has become an important tool in the crystallographic method for studying biomacromolecules (e.g., proteins and peptides). In many instances, cryoprotection at cryogenic temperatures and subsequent data collection are crucial for obtaining a complete data set by overcoming the problem of radiation damage from x-ray beam lines. In addition, cryogenic methods enable crystallographers to handle small crystals, and such methods have become an ideal method for long-term storage of crystals without losing diffraction quality. Cryoprotectants provide means for protecting macromolecular crystals from the destructive effects of ice formation during the freezing process. Cryoprotection generally involves immersing crystals in a solution that forms amorphous glass (i.e., vitrification) and rapidly cooling in liquid nitrogen simultaneously. The ideal cryoprotectant for crystallography should be efficient (i.e., cryoprotectants reach effective results at low concentrations). Currently available cryoprotectants are not efficient. Therefore, if the cryoprotectant concentration is too low, crystal ice will be formed during the experiment, which causes background interference. If the cryoprotectant concentration is too high, the beam energy can cause the instant melting of the crystal structure, causing low-quality data to affect subsequent structural analysis. For example, the state-of-the-art cryoprotectant for X-ray crystallography applications requires the use of 20% ethylene glycol to prevent ice crystal formation at the storage temperature of the crystallized protein. During X-ray data collection, the ethylene glycol heats and the crystals melt, preventing further data collection. For additional information, see, e.g., Garman et al., J. Appl. Cryst. 30:211 (1997).

[0262] In some embodiments, the peptoid polymers and / or salts thereof described herein, the peptoid-peptide hybrids and / or salts thereof described herein, the cryoprotectant fluids described herein, or combinations thereof reduce the dissolution of crystals during X-ray data collection. In some embodiments, the peptoid polymers and / or salts thereof, the peptoid-peptide hybrids and / or salts thereof, the cryoprotectant fluids, or combinations thereof reduce background scattering.

[0263] Biological samples and macromolecules suitable for cryoprotection according to the compositions and methods of the present invention can be from any kingdom (e.g., animal kingdom (including but not limited to humans and livestock animals), plant kingdom, fungi (including but not limited to mushrooms), protists, archaea / archaea, and bacteria / eubacteria).

[0264] On the other hand, the present invention provides beauty care products. In some embodiments, the beauty care product comprises a peptoid polymer as described herein and / or a salt thereof, a peptoid-peptide hybrid as described herein and / or a salt thereof, a cryoprotectant as described herein, or a combination thereof. In some embodiments, the beauty care product is a skin care product. In some embodiments, the skin care product is topically applied. Typical formulations for topical products include creams, serums, ointments, sprays, lotions, and patches.

[0265] On the other hand, the present invention provides antifreeze products, such as deicing or ice suppression products. In some embodiments, antifreeze products include peptoid polymers as described herein and / or its salt, peptoid-peptide hybrids as described herein and / or its salt, cryoprotectant as described herein or its combination. In some embodiments, antifreeze products are used to prevent, suppress or delay the formation of ice on or inside an object, and the object includes but is not limited to general machinery and electrical equipment. In some embodiments, antifreeze products prevent, suppress or delay the formation of ice on or inside an aircraft or its parts, unmanned aerial vehicles, cables, automobiles or their parts (for example, automobile engines), gear systems, brake systems, windows, sprinkler systems, gas pipelines or power cables (for example, including power lines). In other examples, antifreeze products are used as kinetic hydrate inhibitors. In some embodiments, antifreeze products further include ethylene glycol, methanol, propylene glycol, glycerine or its combination.

[0266] On the other hand, the present invention provides frozen food. In some embodiments, the frozen food comprises peptoid polymers and / or salts thereof as described herein, peptoid-peptide hybrids and / or salts thereof as described herein, cryoprotectant fluids as described herein, or combinations thereof. In some embodiments, the frozen food is selected from the group consisting of ice cream, yogurt, seafood, fruit, and meat products. In some embodiments, the frozen food further comprises propylene glycol.

[0267] E. Cryopreservation Protocol

[0268] The compositions and methods described herein are applicable to any number of cryopreservation schemes. As a non-limiting example, the compositions and methods of the present invention can be used for cryopreservation during supercooling to high sub-zero temperatures (e.g., 0°C to -20°C). In the field of organ transplantation, organs are usually cooled on ice (e.g., to 0-4°C), which limits the transplantation window to about 10 hours. By using a cryoprotectant containing a standard small molecule CPA for ex vivo machine perfusion, organs can be preserved for up to 96 hours at a temperature of -6°C. Although it is expected that the cryopreservation temperature will be further reduced to below -6°C, which will extend the possible cryopreservation time, this cannot be done because the high concentration of standard CPA required to further reduce the temperature causes irreversible organ damage due to CPA-related toxicity. For more information, see, e.g., Uygun K et al., Nat. Protoc. 10(3): 484-94(2015). Using ex vivo perfusion methods or otherwise contacting biological samples (e.g., organs and tissues) or macromolecules with peptoid polymers and / or salts thereof, peptoid-peptide hybrids and / or salts thereof, and / or cryoprotectant solutions as described herein, is useful for supercooling to high subzero temperatures, allowing cryopreservation at lower temperatures for longer periods of time than currently available options. Other suitable applications of the present invention to supercooling to high subzero temperatures will be readily apparent to those skilled in the art.

[0269] As another non-limiting example, the compositions and methods of the present invention can be used for cryopreservation during a freezing protocol (e.g., -20°C to -196°C). The freezing protocol is usually performed at a controlled rate (sometimes referred to as slow freezing) during at least a portion of the temperature reduction. For example, a biological sample or macromolecule can be contacted with a peptoid polymer and / or a salt thereof as described herein, a peptoid-peptide hybrid as described herein and / or a salt thereof, and / or a cryoprotectant as described herein, and the temperature is reduced at a controlled rate (e.g., reduced at a rate of 1°C per minute) until the desired temperature is reached. Alternatively, the temperature can be reduced at a controlled rate until the desired temperature (e.g., between -80°C and -180°C) is reached, and then the sample or macromolecule can be quickly frozen (e.g., by immersing the sample or macromolecule in liquid nitrogen or placing the sample or macromolecule on liquid nitrogen). The peptoid polymer and / or salts thereof, peptoid-peptide hybrids and / or salts thereof, and / or cryoprotectant fluid may be contacted with the cryopreserved sample or macromolecule at any time during the protocol, provided that it is prior to the formation of ice crystals that disrupt the preserved sample or macromolecule.

[0270] As another non-limiting example, the compositions and methods of the present invention can be used in low temperature freezing protocols (e.g., -90°C to -196°C). For example, a biological sample or macromolecule can be contacted with a peptoid polymer and / or a salt thereof as described herein, a peptoid-peptide hybrid as described herein and / or a salt thereof, and / or a cryoprotectant as described herein, and then inserted into liquid nitrogen or a stream of liquid nitrogen vapor to rapidly freeze the sample without forming ice crystals. There is no ice lattice, so the water in the sample or macromolecule is in an amorphous or glassy state. Therefore, destructive ice is not formed.

[0271] Those skilled in the art will readily appreciate that the concentration and composition of the peptoid polymers, peptoid polymer salts, peptoid-peptide hybrids, peptoid-peptide hybrid salts, and cryoprotectant solutions described herein may vary depending on the specific biological samples and / or macromolecules being cryopreserved and the specific cryopreservation protocol employed.

[0272] F. Screening Methods

[0273] In a related aspect, the invention provides methods for screening the activity of peptoid polymers and / or salts thereof, peptoid-peptide hybrids and / or salts thereof, and / or cryoprotectant solutions.

[0274] In one embodiment, polarized light microscopy is used to detect ice crystal formation to screen whether peptoid polymers and / or their salts, peptoid-peptide hybrids and / or their salts, and / or cryoprotectants can lower the freezing point of water. Polarized light microscopy is an optical microscopy technique that uses polarized light as a light source. Image contrast arises from the interaction of plane polarized light with birefringent or doubly-refracting substances to produce two separate wave components that are each polarized in mutually perpendicular planes. The speeds of these components, called ordinary wavefronts and extraordinary wavefronts, are different and vary with the direction of propagation through the sample. After leaving the sample, the light components become out of phase, but when they pass through the analyzer, they are recombined with constructive and destructive interference. The interference produces a detectable contrast in the sample. Ice crystal formation is easily detected using this technique because ice crystals are birefringent substances. In a standard experiment, a sample comprising a peptoid polymer and / or a salt thereof, a peptoid-peptide hybrid and / or a salt thereof, and / or a cryoprotectant is cooled to a desired temperature for a desired period of time. One or more samples are placed under a polarized light microscope at the desired temperature and visually inspected for ice crystal formation.

[0275] In one embodiment, differential scanning calorimetry is used to quantitatively measure the thermal hysteresis activity to screen whether peptoid polymers and / or their salts, peptoid-peptide hybrids and / or their salts, and / or cryoprotective solutions can reduce the freezing point of aqueous solutions. Differential scanning calorimetry is a thermal analysis technique in which the difference in the amount of heat required to raise a sample and a reference sample is measured as a function of temperature. When a physical transition such as a phase change occurs, more or less heat will be required to flow to the sample than the reference sample to maintain both at the same temperature. The temperature difference between the phase change of the reference sample and the sample reports the ability of the sample to reduce or inhibit ice crystal formation at temperatures below 0°C. In a standard experiment, a sample comprising a peptoid polymer and / or its salt, a peptoid-peptide hybrid and / or its salt, and / or a cryoprotective solution is compared with a reference sample lacking a peptoid polymer and / or its salt, a peptoid-peptide hybrid and / or its salt, and / or a cryoprotective solution.

[0276] G. Cell viability assay to test activity

[0277] In a related aspect, the invention provides a cell viability assay to test the ability of the peptoid polymers of the invention and / or their salts, the peptoid-peptide hybrids of the invention and / or their salts, and / or the cryoprotectant solutions of the invention to maintain cell viability (e.g., after storage) at reduced temperatures.

[0278] In some embodiments, a PCR product provided by Thermo Fisher Scientific, Inc. is used. Cell viability assay protocol was used to test cell viability. Briefly, is the trade name for resazurin (7-hydroxy-3H-phenoxazine-3-one 10-oxide), a non-toxic, cell-permeable compound that is blue and nearly non-fluorescent. Once inside the cell, resazurin is reduced to the red, highly fluorescent compound resorufin. Living cells continuously convert resazurin to resorufin, increasing the overall fluorescence and color of the culture medium surrounding the cells. Inactive cells cannot convert resazurin to resorufin; therefore, the overall fluorescence and color of the culture medium surrounding the cells are indicators of the relative amount of living cells in the sample. In a standard experiment, cells are mixed with a peptoid polymer and / or a salt thereof, a peptoid-peptide hybrid and / or a salt thereof, and / or a cryoprotectant in any suitable container. The mixture is then cooled to a desired temperature below 0°C and maintained for a desired time. The cells are then returned to ambient temperature and added Reagents, incubation and measurement according to Thermo Fisher protocol. Typically, the sample is measured at wavelength Relative fluorescence at 400 nm was used as a direct readout to determine cell viability.

[0279] In some embodiments, the LIVE / for mammalian cells provided by Thermo Fisher Scientific, Inc. is used. The Viability / Cytotoxicity Kit is used to test cell viability. The kit uses two indicator molecules: Calcein AM and Ethidium homodoimer-1 (EthD-1). Viable cells are characterized by ubiquitous intracellular esterase activity, which is measured by enzymatic conversion of the virtually non-fluorescent, cell-permeable Calcein AM to the highly fluorescent Calcein. The polyanionic dye Calcein is well retained within viable cells, where it produces a strong, uniform green fluorescence. In contrast, EthD-1 enters cells with membrane damage and experiences a 40-fold enhancement in fluorescence upon binding to nucleic acids, resulting in bright red fluorescence in dead cells. Of note, EthD-1 is excluded by the intact cell membrane of live cells, so determination of live and dead cells is readily discriminative. Calcein and EthD-1 can be simultaneously observed with a conventional fluorescein longpass filter. Alternatively, the fluorescence from these dyes can also be observed separately; calcein can be observed with a standard fluorescein bandpass filter, and EthD-1 can be observed with propidium iodide or Texas EthD-1 is observed through a filter that filters the dye. In a standard experiment, cells are mixed with a peptoid polymer and / or a salt thereof, a peptoid-peptide hybrid and / or a salt thereof, and / or a cryoprotectant in any suitable container. The mixture is then cooled to a desired temperature below 0°C, maintained at that temperature for a desired time, and then returned to ambient temperature. Subsequent steps involving the addition of Calcein AM and EthD-1 reagents and measuring the test results are performed as described in the Thermo Fisher protocol. Typically, a direct readout of cell viability is determined by measuring the relative fluorescence at the above-mentioned wavelengths of the two reagents.

[0280] In some embodiments, cell viability is tested using the MTT assay. The MTT assay is a colorimetric cell viability and proliferation assay that relies on the reduction of yellow tetrazolium MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) to insoluble formazan with purple color. Tetrazolium dye reduction depends on NAD(P)H-dependent oxidoreductases that are primarily located in the cytoplasmic compartment of metabolically active cells. The MTT assay can be obtained from, for example, ATCC (www.atcc.org) or Sigma-Aldrich (www.sigmaaldrich.com). In a standard experiment, cells are mixed with peptoid polymers and / or salts thereof, peptoid-peptide hybrids and / or salts thereof, and / or cryoprotectant in any suitable container. The mixture is then cooled to a desired temperature below 0°C and maintained for a desired time. The cells are then returned to ambient temperature, and the MTT reagent is added, incubated, and measured according to ATCC or Sigma-Aldrich protocols. Typically, the absorbance of the converted dye is measured at a wavelength of 570 nm with background subtraction at 630-690 nm.

[0281] IV. Examples

[0282] The following examples are provided to illustrate, but not to limit the claimed invention.

[0283] Example 1. Peptoid-mediated inhibition of ice crystal formation

[0284] This example illustrates the ice crystal inhibition properties of N-substituted peptoid polymers and peptoid-peptide hybrids at temperatures below 0°C.

[0285] Capillary assay

[0286] In this experiment, four aqueous samples were prepared in capillaries containing MilliQ pure water. One sample contained only water, and another sample contained 160mM ethylene glycol (EG). The other two samples each contained 9mM of a peptoid polymer. One peptoid polymer sample contained a peptoid polymer called "Compound 1", and the other sample contained a peptoid polymer called "Compound 10". The sequences of the peptoid polymers are as follows: Compound 1: Nsb-Nsb-Nhp-Nsb-Nsb-Nhp-Nsb-Nsb-Nhp-Nsb (SEQ ID NO: 1); Compound 10: Nhp-Nhp-Nhp-Nhp-Nhp-Nhp-Nhp-Nhp-Nhp-Nhp-Nhp (SEQ ID NO: 2).

[0287] The chemical structures of these compounds are provided in Table 2.

[0288] After sample preparation, all samples were slowly cooled on a Peltier cooling stage and incubated at -20°C. After one hour, the samples were removed and immediately photographed using a digital camera connected to a 180x stereo zoom microscope ( Figure 2A ). The water and EG samples showed obvious ice crystal formation, although the EG sample showed less ice formation than the water-only sample. In contrast, none of the samples containing the peptoid polymer compound showed obvious ice crystal formation. Figure 2B Normalized data are shown. Notably, the EG sample containing CPA concentration approximately 18-fold higher than the peptoid sample still showed significant ice formation, whereas the peptoid sample did not.

[0289] Crystal X-ray diffraction determination

[0290] In order to increase the throughput of library analysis, crystal X-ray diffraction (XRD) techniques were used to evaluate ice crystal formation. For these experiments, compounds named "Compound 2", "Compound 8", "Compound 10", "Compound 11", "Compound 12", "Compound 13", and "Compound 58" were tested. Compounds 2, 8, 10, 11, 12, and 13 are peptoid polymers, the structures of which are given in Table 2. Compound 58 is a peptoid-peptide hybrid, the structure of which is given in Table 10. Compound 58 is similar to Compound 12 except that an arginine amino acid is appended to the N-terminus.

[0291] For these experiments, EG concentrations between 15% and 30% (v / v) were used. Typically, EG, DMSO, and other cryoprotectants were used at concentrations of 35-40% (v / v) during XRD sample analysis to vitrify the solution and avoid diffraction interference from ice crystals. Peptoids and peptoid-peptide compounds were used at concentrations of 1 and 5 mg / mL. Figure 3A , 3B , and 3C show exemplary XRD data under conditions of complete vitrification, partial vitrification in the presence of cubic ice, and freezing (cubic ice crystals), respectively. Figures 4A-4G and Figure 5A-5G The XRD data of compounds 8, 10, 11, 12, 13, and 58 are provided in . Figure 3D Ice ring fractions for various EG concentrations and two concentrations of compounds 2, 8, and 12 are provided.

[0292] Several mixtures of the test solution sample set showed strong anti-icing effects. Figure 3D Experimental results for some peptoid polymer solutions compared to EG are shown. "Ice Ring 1" and "Ice Ring 2" refer to ice formation scores ranging between 0 (no ice formation) and 15 (large ice formation). Compounds 2, 12, and 8, etc. significantly reduce the necessary EG concentration while preventing ice formation.

[0293] The sample containing compound 12 at a concentration of 5 mg / mL (0.5% (w / v)) and EG at a concentration of 17.5% (v / v) in water was ice-free after rapid freezing. This particular mixture was found to completely eliminate all ice formation in multiple rapid freezing tests in a stream of liquid nitrogen vapor ( Figure 3A ) and is much better than the standard solution of 30% EG ( Figure 3B ). In the figure, black dots and circles represent ice crystals. The anti-ice effect is 500 times stronger than that of EG at the same molar concentration and, without being bound by any particular theory, suggests a non-colligative mechanism for anti-ice, which is the mechanism used by natural antifreeze proteins.

[0294] Larger volume measurements

[0295] To test the usefulness of the peptoid polymer on a larger scale, experiments were performed using solution volumes similar to those used for standard egg and stem cell storage. For these experiments, two samples, one containing only 22.5% EG and buffer, and the other containing 22.5% EG and 5 mg / ml (0.5% w / v) compound 12 and buffer, were flash frozen in liquid nitrogen. Fig. 6A As shown, the Compound 12 solution showed complete vitrification after being removed from liquid nitrogen, without immediate ice formation, while the control solution was obviously frozen, producing a large number of white ice crystals. Rewarming of the solution in a 37°C water bath resulted in an unexpected and beneficial effect. The Compound 12 solution underwent devitrification in less than 2 seconds upon rewarming ( Figure 6B , right), while ice cubes were seen floating in the control sample after 20 seconds ( Figure 6B , left). Condensation is seen on each sample tube because the sample tubes are actually still well below room temperature. This result indicates that compound 12 acts as an active deicing agent during thawing.

[0296] In addition, after placing 100 μL of the sample in a -20°C refrigerator overnight, it was found that the compound 12 solution was not frozen ( Figure 6C , right). This result indicates that the peptoid polymer provides the ability to preserve samples for long periods of time at temperatures below 0°C without any ice formation. In addition, these experiments show that ice-free conditions can be achieved by introducing peptoid polymers at -20°C and near vitrification to -80°C with cryopreservation, or by supercooling methods, thereby significantly reducing the critical concentration of osmotic CPA and alleviating cryopreservation toxicity.

[0297] As shown here, it was found that the formulation of compound 12 prevented ice formation in sub-milliliter volumes during vitrification. In fact, the solution was able to remain completely thawed at -20°C and was also able to vitrify when rapidly frozen at -196°C. Currently, standard human oocyte preservation techniques for in vitro fertilization are limited to solution volumes less than 5uL (typically 0.5 to 2.5 μL) while using cryoprotectant concentrations of more than 50%. Therefore, compound 12 can prevent ice formation in practical volumes with very little cryoprotectant, which makes it useful, for example, for preserving human oocytes for in vitro fertilization.

[0298] Example 2. Cytotoxicity and cryopreservation screening

[0299] This example demonstrates that peptoid polymers have little cytotoxicity when compared to existing compounds and can achieve excellent cryopreservation while reducing the amount of CPA required and thus reducing CPA-associated toxicity.

[0300] Cytotoxicity assay

[0301] To demonstrate the safety of the cryoprotectant compositions, a cell-based high-throughput cytotoxicity assay was developed using the HEK 293 cell line, a robust and robust stem cell line grown in tissue culture from human embryonic kidney cells.

[0302] Solutions were prepared in 96- and 384-well plates using a Tecan Genesis robotic workstation. The solution contained culture medium, buffer, cryoprotectant composition (compound 12) or DMSO. The solution was adjusted to the desired pH. Serial dilutions were performed to obtain solutions containing various concentrations of compound 12 and DMSO. Control experiments were performed using culture medium only.

[0303] For these experiments, cells were seeded at low density (i.e., 10% confluence), exposed to solutions containing compound 12 or DMSO, and placed in a 37°C incubator. Cells were grown until control cells treated with empty vector alone approached 70% confluence (usually about 3 to 5 days). The cytotoxicity of the compounds was assessed by MTT assay.

[0304] like Figure 7 As shown, the toxicity of compound 12 did not deviate significantly from that of the medium alone when analyzed by the MTT assay. On the other hand, DMSO did not allow rewarming survival for an extended period of time at any concentration above 0.5% (v / v). Notably, compound 12 did not show toxicity at a concentration (0.5% w / v) at which it could prevent ice formation in non-biological samples, and did not show significant toxicity at a concentration 4-fold higher than this concentration.

[0305] These results indicate that the cryoprotectant composition is effective in protecting against ice even at concentrations where DMSO toxicity significantly reduces cell survival.

[0306] Cryopreservation assay

[0307] Very simple solutions were used to carry out the initial cryopreservation assays with and without the addition of compound 12 to minimize confounding external factors. For the first set of experiments, two sample solutions were prepared. The first sample solution contained a simple buffer and ethylene glycol (EG) at a concentration of 22.5% (v / v), and the second sample solution contained a simple buffer, EG (22.5% (v / v)), and 5 mg / mL (0.5% (w / v)) of compound 12.

[0308] HEK 293 cells were grown until 70% confluent and then treated with trypsin to remove adhesion proteins and produce free floating cells. Cells were counted using a hemocytometer and the sample cell concentration was adjusted to a final concentration of 10,000 cells per microliter. The cells were compressed into a tight mass by centrifugation, after which each sample was mixed with one of 20 μL of sample solution. The sample was then quickly frozen by immersing it in liquid nitrogen and then rewarmed in a 37°C water bath. After the freeze-thaw process, the cells were suspended in a 400x volume of culture medium to recover. The positive control sample was treated with culture medium at 37°C and the freeze-thaw process was not performed. The negative control sample was treated with culture medium only during the freeze-thaw process. After recovery, the cells were stained with calcein AM for 30 minutes and the cell viability was measured using a fluorescent plate reader.

[0309] like Figure 8 As shown, the addition of compound 12 greatly improved cell survival and demonstrated the ability of the compound to cryopreserve cells. It was observed that samples containing compound 12 achieved complete vitrification without ice formation during the freezing process. In addition, compared with samples lacking compound 12, the vitrification process was crossed faster.

[0310] A second set of experiments was performed to evaluate the cryopreservability of a formulation containing 5 mg / mL of Compound 12 plus a mixture of a diol, a disaccharide, and a general buffer. Post-thaw survival after vitrification in liquid nitrogen was evaluated as described above. Fig. 9 As can be seen in the Figure 3, this preparation achieved nearly 100% (i.e., 98%) post-thaw survival of cells, which is similar to the control group that was not exposed to the freezing treatment. Cell morphology and fluorescent signal appear to be the same as the non-frozen control, indicating that the cells suffered little damage during the experiment.

[0311] As part of a second set of experiments, the cryopreservation ability of the formulation was compared with two known cryopreservation agents. VS2E is a DMSO-free and serum-free solution containing a non-chemically defined polymer (see, e.g., Nishigaki et al., Int. J. Dev. Biol. 55:3015-311 (2011)), and M22 is a cryopreservation agent derived from 21 st Organ vitrification solution obtained from Century Medicine. Fig. 9 The formulation containing compound 12 was shown to achieve excellent cryopreservation, as the cell viability of VS2E and M22 was 72% and 51%, respectively. It should be noted that for the M22 sample, background fluorescence may have affected the accuracy of this result, as the count of live cells in the image indicated much less than 51% cell viability.

[0312] The peptoid polymers are very effective in preventing ice formation in solutions containing significantly reduced amounts of ethylene glycol. Specifically, at a 20 uL scale for preserving various types of cells, low concentrations of the composition (e.g., 0.5% (w / v)) are sufficient to prevent ice growth during vitrification and keep the solution in a liquid, ice-free state.

[0313] In summary, these results show that peptoid polymers can achieve excellent cryopreservation and reduce the amount of CPA required, thereby reducing the cytotoxicity associated with CPA. The superior properties of peptoid polymers are particularly useful for the treatment of particularly sensitive cell lines and / or when cells need to be cultured for longer periods of time.

[0314] Example 3. Vitrification efficacy test

[0315] As another means of increasing the throughput of library analysis, vitrification screening experiments were performed to evaluate anti-ice potency. For these experiments, compounds named "Compound 11", "Compound 12", "Compound 25", "Compound 26", "Compound 27", "Compound 28", "Compound 60", "Compound 62", "Compound 63", "Compound 64", "Compound 69", "Compound 70", "Compound 71", "Compound 72", "Compound 73", "Compound 74", "Compound 75", "Compound 76", "Compound 77", "Compound 78", "Compound 81", "Compound 82", "Compound 83", "Compound 84" and "Compound 85" were tested. Each compound is a peptoid polymer, and its structure is provided in Tables 2, 5, and 11.

[0316] For these experiments, 20 μL samples were flash frozen by liquid immersion in liquid nitrogen. Ethylene glycol (EG) concentrations between 5% and 25% (v / v) were used. The peptoid polymer compound was used at a concentration of 5 mg / mL in the formulation. Table 12 lists the minimum concentration of EG required to completely vitrify a 20 μL sample with a peptoid polymer in the formulation, referred to as the "critical EG concentration." Samples of formulations without the peptoid polymer were only able to achieve vitrification at 20% or more of EG. The target peptoid polymer can achieve vitrification in a formulation containing EG when the concentration of EG is less than 20%.

[0317] Several peptoids were shown to have critical EG concentrations below 20%, demonstrating their usefulness as anti-ice agents. Compounds 60, 62, 82, 83, 84, and 85 required significantly less EG to achieve vitrification. These compounds are therefore able to prevent ice formation in practical volumes, which is useful for cryopreservation of biological materials.

[0318]

[0319] Example 4. Cell viability and toxicity assay

[0320] This example describes a number of experiments performed to evaluate the effects of the compositions of the invention on cell viability, survival, and toxicity.

[0321] Evaluation of the effect of peptoid polymers on the survival rate of K562 cells at -20°C for 3 days

[0322] Several compounds were tested for their ability to increase the viability of cells subjected to supercold storage at a 1% concentration (i.e., 1% peptoid polymer concentration) in an XT formulation. The XT formulation contains HTK buffer, among other components. In particular, the effects of these compounds on the viability of K562 cells stored at -20°C for 3 days were evaluated. The Alamar Blue assay was used to assess cell viability 7 days after thawing of stored cells. Fig.10 As shown, compounds 12, 27, 62, 74, and 76 improved cell viability compared to the XT formulation alone.

[0323] Evaluation of the effect of peptoid polymers on the survival rate of K562 cells at -80°C for 3 days

[0324] Several compounds were tested for their effects on cryopreservation of cells at 1% concentration (i.e., 1% peptoid polymer concentration) in XT formulations. In particular, the effects of these compounds on the viability of K562 cells stored at -80°C for 3 days were evaluated. The Alamar Blue assay was used to assess cell viability 1 day after thawing of stored cells. 10% DMSO formulations and DMEM cell culture medium alone were used as controls. Fig.11As shown, compound 76 produced the greatest improvement in cell viability.

[0325] Evaluation of the effect of peptoid polymers on the survival rate of HEK293 cells at -80°C for 3 days

[0326] Several compounds were tested at 1% concentration (i.e., 1% peptoid polymer concentration) in XT formulations for their effects on cryopreservation of HEK293 cells, an adherent cell line derived from human embryonic kidney cells. In particular, the effects of these compounds on the viability of HEK293 cells stored at -80°C for 3 days were evaluated. The Alamar Blue assay was used to assess cell viability 1 day after thawing of stored cells. 10% DMSO formulations and DMEM cell culture medium alone were used as controls. Fig.12 As shown, compounds 11, 27, 62, 64, and 75 improved cell viability compared to the XT formulation alone.

[0327] Evaluation of cytotoxic effects

[0328] The cytotoxicity of several compounds was evaluated in K562 cell culture using a typical cytotoxicity assay. K562 cells were incubated with peptoid polymers (i.e., compounds 12, 15, or 62 at 1% concentration) at 37°C and 5% CO2 for 24 hours. As a positive control, some cells were incubated without any peptoid polymer (DMEM). As a negative control, cells were incubated with DMSO (DMEM + 10% DMSO). Fig.13 As shown, cells incubated with the peptoid polymer persisted as well as cells incubated in cell culture medium alone (DMEM), but cells incubated with DMSO suffered significant toxicity.

[0329]

[0330]

[0331]

[0332]

[0333]

[0334]

[0335]

[0336]

[0337]

[0338]

[0339]

[0340]

[0341]

[0342]

[0343]

[0344]

[0345]

[0346] V. Exemplary Embodiments

[0347] Exemplary embodiments provided according to the presently disclosed subject matter include, but are not limited to, the claims and the following embodiments:

[0348] 1. A peptoid polymer or a salt thereof comprising subunits, wherein the subunits comprise one or more first hydrophobic peptoid monomers H and one or more first polar peptoid monomers P, which are arranged so that the peptoid polymer has a sequence [H a P b ] n or [P b H a ] n ,in:

[0349] The subscript a indicating the number of consecutive first hydrophobic peptoid monomers in the subunit is between 1 and 10;

[0350] The subscript b indicating the number of consecutive first polar peptoid monomers in the subunit is between 1 and 10; and

[0351] The subscript n, denoting the number of subunits in the peptoid polymer, is between 2 and 50.

[0352] 2. The peptoid polymer or salt thereof according to embodiment 1, further comprising substituents X and Y such that the peptoid polymer has a sequence X-[H a P b ] n -Y or X-[P b H a ] n -Y, where:

[0353] X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C1-8 Alkylamino, -OH, -SH, -NH2, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 carboxyalkyl, and halogen, or

[0354] Optionally, X and Y together form a covalent bond.

[0355] 3. The peptoid polymer or salt thereof according to embodiment 1, wherein the subunit further comprises a second hydrophobic peptoid monomer and / or a second polar peptoid monomer, so that the peptoid polymer has a sequence [H a P b H c P d ] n or [P b H a P d H c ] n ,in:

[0356] The subscript c indicating the number of consecutive second hydrophobic peptoid monomers in the subunit is between 0 and 10;

[0357] The subscript d representing the number of consecutive second polar peptoid monomers in the subunit is between 0 and 10; and

[0358] c and d are not both 0.

[0359] 4. The peptoid polymer or salt thereof according to embodiment 3, further comprising substituents X and Y such that the peptoid polymer has a sequence X-[H a P b H c P d ] n -Y or X-[P b H a P d H c ] n -Y, where:

[0360] X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C1-8 carboxyalkyl, and halogen, or

[0361] Optionally, X and Y together form a covalent bond.

[0362] 5. A peptoid polymer or a salt thereof according to any one of embodiments 1 to 4, further comprising a sequence Z, wherein the sequence Z comprises one or more hydrophobic peptoid monomers and / or one or more polar peptoid monomers, wherein Z is located before the first subunit, after the last subunit, and / or between one or more subunits.

[0363] 6. The peptoid polymer or salt thereof according to embodiment 5, wherein Z comprises one or more hydrophobic peptoid monomers.

[0364] 7. The peptoid polymer or salt thereof according to embodiment 5, wherein Z comprises one or more polar peptoid monomers.

[0365] 8. The peptoid polymer or salt thereof according to embodiment 5, wherein Z comprises one or more hydrophobic peptoid monomers and one or more polar peptoid monomers.

[0366] 9. The peptoid polymer or salt thereof according to any one of embodiments 1 to 8, wherein n is between 2 and 10.

[0367] 10. The peptoid polymer or salt thereof according to any one of embodiments 1 to 9, wherein a is between 1 and 5.

[0368] 11. The peptoid polymer or salt thereof according to any one of embodiments 1 to 10, wherein b is between 1 and 5.

[0369] 12. The peptoid polymer or salt thereof according to embodiment 10 or 11, wherein a is between 1 and 3 and b is between 1 and 3.

[0370] 13. The peptoid polymer or salt thereof according to any one of embodiments 3 to 12, wherein c is between 0 and 5.

[0371] 14. The peptoid polymer or salt thereof according to any one of embodiments 3 to 13, wherein d is between 0 and 5.

[0372] 15. A peptoid polymer or a salt thereof, comprising:

[0373] (a) a subunit comprising two first hydrophobic peptoid monomers H and two first polar peptoid monomers P, and

[0374] (b) two second hydrophobic peptoid monomers located at the C-terminus of the peptoid polymer,

[0375] The arrangement is such that the peptoid polymer has the sequence [H2P2]n H2 or [P2H2] n H2, wherein the subscript n representing the number of subunits in the peptoid polymer is between 1 and 50.

[0376] 16. The peptoid polymer or salt thereof according to embodiment 15, which comprises compound 81.

[0377] 17. The peptoid polymer or salt thereof according to embodiment 15 or 16, further comprising substituents X and Y such that the peptoid polymer has a sequence X-[H2P2] n H2-Y or X-[P2H2] n H2-Y, where:

[0378] X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 carboxyalkyl, and halogen, or

[0379] Optionally, X and Y together form a covalent bond.

[0380] 18. The peptoid polymer or salt thereof according to any one of embodiments 15 to 17, wherein n is between 1 and 10.

[0381] 19. The peptoid polymer or salt thereof according to any one of embodiments 1 to 18, wherein the first and / or second hydrophobic peptoid monomer is independently selected from the group consisting of:

[0382]

[0383]

[0384] wherein the subscript m is the number of repeating units and is between 1 and 10.

[0385] 20. The peptoid polymer or salt thereof according to any one of embodiments 1 to 15 or 17 to 19, wherein none of the polar peptoid monomers comprises an optionally substituted C 1-18 A hydroxyalkyl side chain.

[0386] 21. The peptoid polymer or salt thereof according to any one of embodiments 1 to 20, wherein each of the first and / or second polar peptoid monomers comprises a side chain independently selected from (C 1-6 Alkoxy) (C 1-6 alkylene), (oligo[ethylene glycol]), (4- to 10-membered heterocycloalkyl) (C 1-6 alkylene), and (5- to 10-membered heteroaryl) (C 1-6 alkylene).

[0387] 22. The peptoid polymer or salt thereof according to embodiment 21, wherein (4- to 10-membered heterocycloalkyl) (C 1-6 The term "alkylene" refers to a 4-6 membered heterocyclic ring wherein at least one member is selected from the group consisting of O and N.

[0388] 23. The peptoid polymer or salt thereof according to embodiment 21 or 22, wherein (4- to 10-membered heterocycloalkyl) (C 1-6 Alkylene) includes tetrahydrofuranyl or oxopyrrolidinyl moieties.

[0389] 24. The peptoid polymer or salt thereof according to embodiment 23, wherein the peptoid polymer comprises

[0390]

[0391] 25. The peptoid polymer or salt thereof according to embodiment 24, wherein all polar peptoid monomers are

[0392] 26. The peptoid polymer or salt thereof according to embodiment 24 or 25, wherein the peptoid polymer comprises Compound 63, Compound 76, Compound 86, or Compound 87.

[0393] 27. The peptoid polymer or salt thereof according to embodiment 21, wherein (5- to 10-membered heteroaryl) (C 1-6 The term "alkylene" refers to a 5-6 membered aromatic ring wherein at least one ring member is selected from the group consisting of O and N.

[0394] 28. The peptoid polymer or salt thereof according to embodiment 21 or 27, wherein (5- to 10-membered heteroaryl) (C 1-6 Alkylene) includes a furyl moiety.

[0395] 29. The peptoid polymer or salt thereof according to embodiment 28, wherein the peptoid polymer comprises

[0396]

[0397] 30. The peptoid polymer or a salt thereof according to embodiment 29, wherein the peptoid polymer comprises compound 73.

[0398] 31. The peptoid polymer or salt thereof according to embodiment 21, wherein the side chain comprises a methoxyethyl group.

[0399] 32. The peptoid polymer or salt thereof according to embodiment 31, wherein the peptoid polymer comprises

[0400]

[0401] 33. The peptoid polymer or a salt thereof according to embodiment 32, wherein the peptoid polymer comprises compound 62.

[0402] 34. The peptoid polymer or salt thereof of embodiment 21, wherein the side chain comprises an oligo(ethylene glycol) moiety.

[0403] 35. The peptoid polymer or salt thereof of embodiment 34, wherein the oligo(ethylene glycol) moiety is a 2-(2-(2-methoxyethoxy)ethoxy)ethyl moiety.

[0404] 36. The peptoid polymer or salt thereof according to embodiment 35, wherein the peptoid polymer comprises

[0405]

[0406] 37. The peptoid polymer or a salt thereof according to embodiment 36, wherein the peptoid polymer comprises compound 67.

[0407] 38. The peptoid polymer or salt thereof according to any one of embodiments 1 to 19, wherein the peptoid polymer comprises a polar peptoid monomer having a hydroxyl-containing side chain.

[0408] 39. The peptoid polymer or salt thereof according to any one of embodiments 1 to 19, wherein the first and / or second polar peptoid monomer is independently selected from the group consisting of:

[0409]

[0410]

[0411]

[0412] wherein the subscript m is the number of repeating units and is between 1 and 10.

[0413] 40. A peptoid polymer or a salt thereof, comprising one or more hydrophobic peptoid monomers and one or more polar peptoid monomers, wherein the one or more polar peptoid monomers each comprise a hydrophobic peptoid monomer selected independently from (C 1-6 Alkoxy) (C 1-6 alkylene), (oligo[ethylene glycol]), (4- to 10-membered heterocycloalkyl) (C 1-6 alkylene), and (5- to 10-membered heteroaryl) (C 1-6 alkylene).

[0414] 41. The peptoid polymer or salt thereof according to embodiment 40, wherein the one or more hydrophobic peptoid monomers are each independently selected from the group consisting of:

[0415] wherein the subscript m is the number of repeating units and is between 1 and 10.

[0416] 42. The peptoid polymer or salt thereof according to embodiment 40 or 41, wherein (4- to 10-membered heterocycloalkyl) (C 1-6 The term "alkylene" refers to a 4-6 membered heterocyclic ring wherein at least one member is selected from the group consisting of O and N.

[0417] 43. The peptoid polymer or salt thereof according to any one of embodiments 40 to 42, wherein (4- to 10-membered heterocycloalkyl) (C 1-6 Alkylene) includes tetrahydrofuranyl or oxopyrrolidinyl moieties.

[0418] 44. The peptoid polymer or salt thereof according to embodiment 43, wherein the peptoid polymer comprises

[0419]

[0420] 45. The peptoid polymer or salt thereof according to embodiment 44, wherein all polar peptoid monomers are

[0421] 46. ​​The peptoid polymer or salt thereof according to embodiment 44 or 45, wherein the peptoid polymer comprises Compound 63, Compound 76, Compound 86, or Compound 87.

[0422] 47. The peptoid polymer or salt thereof according to embodiment 40 or 41, wherein (5- to 10-membered heteroaryl) (C 1-6 The term "alkylene" refers to a 5-6 membered aromatic ring wherein at least one ring member is selected from the group consisting of O and N.

[0423] 48. The peptoid polymer or salt thereof according to embodiment 40, 41, or 47, wherein (5- to 10-membered heteroaryl) (C 1-6 Alkylene) includes a furyl moiety.

[0424] 49. The peptoid polymer or salt thereof according to embodiment 48, wherein the peptoid polymer comprises

[0425]

[0426] 50. The peptoid polymer or a salt thereof according to embodiment 49, wherein the peptoid polymer comprises compound 73.

[0427] 51. The peptoid polymer or salt thereof of embodiment 40 or 41, wherein the side chain comprises a methoxyethyl group.

[0428] 52. The peptoid polymer or salt thereof according to embodiment 51, wherein the peptoid polymer comprises

[0429]

[0430] 53. The peptoid polymer or a salt thereof according to embodiment 52, wherein the peptoid polymer comprises compound 62.

[0431] 54. The peptoid polymer of embodiment 40 or 41, wherein the side chain comprises an oligo(ethylene glycol) moiety.

[0432] 55. The peptoid polymer or salt thereof of embodiment 54, wherein the oligo(ethylene glycol) moiety is a 2-(2-(2-methoxyethoxy)ethoxy)ethyl moiety.

[0433] 56. The peptoid polymer or salt thereof according to embodiment 55, wherein the peptoid polymer comprises

[0434]

[0435] 57. The peptoid polymer or a salt thereof according to embodiment 56, wherein the peptoid polymer comprises compound 67.

[0436] 58. The peptoid polymer or salt thereof according to any one of embodiments 40 to 57, further comprising substituents X and Y located at the N-terminus and C-terminus of the peptoid polymer, respectively, wherein:

[0437] X and Y are independently selected from the group consisting of: H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, carboxyl, optionally substituted C1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 carboxyalkyl, and halogen, or

[0438] Optionally, X and Y together form a covalent bond.

[0439] 59. The peptoid polymer or salt thereof according to any one of embodiments 1 to 58, wherein about 10% of the peptoid monomers are hydrophobic.

[0440] 60. The peptoid polymer or salt thereof according to any one of embodiments 1 to 58, wherein about 20% of the peptoid monomers are hydrophobic.

[0441] 61. The peptoid polymer or salt thereof according to any one of embodiments 1 to 58, wherein about 30% of the peptoid monomers are hydrophobic.

[0442] 62. The peptoid polymer or salt thereof according to any one of embodiments 1 to 58, wherein about 40% of the peptoid monomers are hydrophobic.

[0443] 63. The peptoid polymer or salt thereof according to any one of embodiments 1 to 58, wherein about 50% of the peptoid monomers are hydrophobic.

[0444] 64. The peptoid polymer or salt thereof according to any one of embodiments 1 to 58, wherein about 60% of the peptoid monomers are hydrophobic.

[0445] 65. The peptoid polymer or salt thereof according to any one of embodiments 1 to 58, wherein about 70% of the peptoid monomers are hydrophobic.

[0446] 66. The peptoid polymer or salt thereof according to any one of embodiments 1 to 58, wherein about 80% of the peptoid monomers are hydrophobic.

[0447] 67. The peptoid polymer or salt thereof according to any one of embodiments 1 to 58, wherein about 90% of the peptoid monomers are hydrophobic.

[0448] 68. The peptoid polymer or salt thereof according to any one of embodiments 1 to 67, wherein the peptoid polymer forms a helical structure.

[0449] 69. The peptoid polymer or salt thereof according to any one of embodiments 1 to 68, wherein the peptoid polymer reduces or inhibits ice crystal formation at a temperature within about 0°C to about -20°C.

[0450] 70. The peptoid polymer or salt thereof according to any one of embodiments 1 to 68, wherein the peptoid polymer reduces or inhibits ice crystal formation at a temperature within about -20°C to about -40°C.

[0451] 71. The peptoid polymer or salt thereof according to any one of embodiments 1 to 68, wherein the peptoid polymer reduces or inhibits ice crystal formation at a temperature of about -20°C.

[0452] 72. The peptoid polymer or salt thereof according to any one of embodiments 1 to 68, wherein the peptoid polymer reduces or inhibits ice crystal formation at a temperature within about -40°C to about -200°C.

[0453] 73. A peptoid polymer or a salt thereof according to any one of embodiments 1 to 72, wherein the salt of the peptoid polymer is selected from the group consisting of hydrochloride, acetate, sulfate, phosphate, maleate, citrate, methanesulfonate, nitrate, tartrate, gluconate, and combinations thereof.

[0454] 74. The peptoid polymer or salt thereof according to any one of embodiments 1 to 73, wherein the peptoid polymer is not a peptoid polymer described in Tables 2-9.

[0455] 75. A peptoid-peptide hybrid or a salt thereof, comprising a peptoid polymer or a salt thereof according to any one of embodiments 1 to 74 and one or more amino acids, wherein the one or more amino acids are located at one or both ends of the peptoid polymer and / or between one or more subunits.

[0456] 76. The peptoid-peptide hybrid or a salt thereof according to embodiment 75, wherein the peptoid-peptide hybrid is not the peptoid-peptide hybrid described in SEQ ID NO: 3, 4, 5, or 6.

[0457] 77. The peptoid-peptide hybrid or a salt thereof according to embodiment 75 or 76, wherein one or more amino acids are selected from the group consisting of alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, arginine, lysine, leucine, methionine, asparagine, proline, glutamine, serine, threonine, valine, tryptophan, tyrosine, and combinations thereof.

[0458] 78. The peptoid-peptide hybrid or a salt thereof according to embodiment 77, wherein the one or more amino acids are selected from the group consisting of isoleucine, leucine, serine, threonine, alanine, valine, arginine, and combinations thereof.

[0459] 79. The peptoid-peptide hybrid of any one of embodiments 75 to 78, wherein the peptoid-peptide hybrid salt is selected from the group consisting of hydrochloride, acetate, sulfate, phosphate, maleate, citrate, methanesulfonate, nitrate, tartrate, gluconate, and combinations thereof.

[0460] 80. A cryoprotectant solution comprising the peptoid polymer or a salt thereof according to any one of embodiments 1 to 74, the peptoid-peptide hybrid or a salt thereof according to any one of embodiments 75 to 79, or a combination thereof.

[0461] 81. The cryoprotectant according to embodiment 80, further comprising a compound selected from the group consisting of: an ionic substance, a permeating cryoprotectant, a non-permeating cryoprotectant, an antioxidant, a cell membrane stabilizing compound, a water channel protein or other channel forming compound, an alcohol, a sugar, a sugar derivative, a non-ionic surfactant, a protein, dimethyl sulfoxide (DMSO), polyethylene glycol (PEG), polypropylene glycol (PPG), Polyvinyl pyrrolidone, polyvinyl alcohol, hyaluronic acid, formamide, natural or synthetic hydrogels, and combinations thereof.

[0462] 82. The cryoprotectant solution of embodiment 81, wherein the alcohol is selected from the group consisting of propylene glycol, ethylene glycol, glycerol, methanol, butylene glycol, adonitol, ethanol, trimethylene alcohol, diethylene glycol, polyethylene oxide, erythritol, sorbitol, xylitol, polypropylene glycol, 2-methyl-2,4-pentanediol (MPD), mannitol, inositol, dithiodiol, 1,2-propylene glycol, and combinations thereof.

[0463] 83. The cryoprotectant solution of embodiment 81, wherein the sugar is a monosaccharide.

[0464] 84. The cryoprotectant solution of embodiment 83, wherein the monosaccharide is selected from the group consisting of glucose, 3-O-methyl-D-glucopyranose, galactose, arabinose, fructose, xylose, mannose, and combinations thereof.

[0465] 85. The cryoprotectant solution of embodiment 81, wherein the sugar is a disaccharide.

[0466] 86. The cryoprotectant solution of embodiment 85, wherein the disaccharide is selected from the group consisting of sucrose, trehalose, lactose, maltose, and combinations thereof.

[0467] 87. The cryoprotectant solution of embodiment 81, wherein the sugar is a polysaccharide.

[0468] 88. The cryoprotectant solution of embodiment 87, wherein the polysaccharide is selected from the group consisting of raffinose, dextran, and combinations thereof.

[0469] 89. The cryoprotectant solution of embodiment 81, wherein the PEG or PPG has an average molecular weight of less than about 3,000 g / mol.

[0470] 90. The cryoprotectant solution of embodiment 81, wherein the PEG or PPG has an average molecular weight between about 200 g / mol and 400 g / mol.

[0471] 91. The cryoprotectant solution of embodiment 81, wherein the protein is selected from the group consisting of ovalbumin, bovine serum albumin, human serum albumin, gelatin, and combinations thereof.

[0472] 92. The cryoprotectant solution of embodiment 81, wherein the natural or synthetic hydrogel comprises chitosan, hyaluronic acid, or a combination thereof.

[0473] 93. The cryoprotectant solution of embodiment 81, wherein the nonionic surfactant is selected from the group consisting of polyoxyethylene lauryl ether, polysorbate 80, and combinations thereof.

[0474] 94. A method for preserving tissue, organ, or cell, the method comprising contacting the tissue, organ, or cell with a peptoid polymer or a salt thereof according to any one of embodiments 1 to 74, a peptoid-peptide hybrid or a salt thereof according to any one of embodiments 75 to 79, a cryoprotectant solution according to any one of embodiments 80 to 93, or a combination thereof.

[0475] 95. A method according to embodiment 94, wherein the peptoid polymer or its salt, the peptoid-peptide hybrid or its salt, the cryoprotectant fluid, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature within the range of about 0°C to about -20°C.

[0476] 96. A method according to embodiment 94, wherein the peptoid polymer or its salt, the peptoid-peptide hybrid or its salt, the cryoprotectant fluid, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature within the range of about -20°C to about -40°C.

[0477] 97. A method according to embodiment 94, wherein the peptoid polymer or a salt thereof, the peptoid-peptide hybrid or a salt thereof, the cryoprotectant fluid, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature of about -20°C.

[0478] 98. A method according to embodiment 94, wherein the peptoid polymer or its salt, the peptoid-peptide hybrid or its salt, the cryoprotectant, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature within the range of about -40°C to about -200°C.

[0479] 99. The method of any one of embodiments 94 to 98, wherein the tissue is a bioengineered tissue.

[0480] 100. The method of any one of embodiments 94 to 99, wherein the peptoid polymer or a salt thereof, the peptoid-peptide hybrid or a salt thereof, or a combination thereof is present in an amount between about 100 nM and about 1,000 mM.

[0481] 101. The method of any one of embodiments 94 to 100, wherein the tissue, organ, or cell is selected from the group consisting of heart, liver, lung, kidney, pancreas, intestine, thymus, cornea, neural cells, platelets, sperm cells, oocytes, genital urinary cells, embryonic cells, stem cells, human pluripotent stem cells, hematopoietic stem cells, lymphocytes, granulocytes, immune system cells, bone cells, primary cells, organoids, and combinations thereof.

[0482] 102. A method for preserving biological macromolecules, the method comprising contacting the biological macromolecules with a peptoid polymer or a salt thereof according to any one of embodiments 1 to 74, a peptoid-peptide hybrid or a salt thereof according to any one of embodiments 75 to 79, a cryoprotectant solution according to any one of embodiments 80 to 93, or a combination thereof.

[0483] 103. The method of embodiment 102, wherein the biomacromolecule is selected from the group consisting of nucleic acids, amino acids, proteins, isolated proteins, peptides, lipids, complex structures, and combinations thereof.

[0484] 104. A beauty care product comprising the peptoid polymer or a salt thereof according to any one of embodiments 1 to 74, the peptoid-peptide hybrid or a salt thereof according to any one of embodiments 75 to 79, the cryoprotectant solution according to any one of embodiments 80 to 93, or a combination thereof.

[0485] 105. An antifreeze product comprising the peptoid polymer or salt thereof according to any one of embodiments 1 to 74, the peptoid-peptide hybrid or salt thereof according to any one of embodiments 75 to 79, the cryoprotectant solution according to any one of embodiments 80 to 93, or a combination thereof.

[0486] 106. The anti-icing product of embodiment 105, wherein the anti-icing product is a de-icing or ice suppression product for preventing, inhibiting, or delaying the formation of ice on or within an object.

[0487] 107. The antifreeze product of embodiment 106, wherein the object is selected from the group consisting of an aircraft or parts thereof, a gas pipeline, a window, an electrical device, a drone, a cable, a power line, a mechanical device, an automobile engine, a gear system, and a brake system.

[0488] 108. A frozen food comprising the peptoid polymer or salt thereof according to any one of embodiments 1 to 74, the peptoid-peptide hybrid or salt thereof according to any one of embodiments 75 to 79, the cryoprotectant solution according to any one of embodiments 80 to 93, or a combination thereof.

[0489] 109. The frozen food of embodiment 108, wherein the frozen food is selected from the group consisting of ice cream, yogurt, seafood, fruit, and meat products.

[0490] It should be understood that the examples and embodiments described herein are for illustrative purposes only, and that various modifications or changes in view thereof will be suggested to those skilled in the art and are included within the spirit and scope of the present application and the scope of the appended claims. All publications, patents, patent applications, and serial accession numbers cited herein are incorporated herein by reference in their entirety for all purposes.

[0491] Informal sequence listing

[0492]

[0493]

[0494]

Claims

1. A peptoid polymer or a salt thereof having the sequence [H2P2] n , [P2H2] n 、X-[H2P2] n -Y, X-[P2H2] n -Y, [H2P2] n H2, [P2H2] n H2, [H2P2] n P2 or [P2H2] n P2, where the subscript n is between 2 and 5, Wherein H is a hydrophobic peptoid monomer selected from where the subscript m is the number of repeating units and is between 1 and 10, wherein P is a polar peptoid monomer selected from a tetrahydrofuranyl moiety or an oxopyrrolidyl moiety, and wherein X and Y are independently selected from H, optionally substituted C 1-8 Alkyl, optionally substituted C 1-8 Acyl, optionally substituted C 1-8 Alkylamino, -OH, -SH, -NH2, secondary amine, tertiary amine, carboxyl, optionally substituted C 1-8 Hydroxyalkyl, optionally substituted C 1-8 Alkylamino, optionally substituted C 2-8 Alkylthio, optionally substituted C 1-8 Carboxyalkyl or halogen.

2. The peptoid polymer or salt thereof according to claim 1, wherein P is 3. The peptoid polymer or salt thereof according to claim 2, wherein P is 4. The peptoid polymer or a salt thereof according to claim 2 or 3, wherein the peptoid polymer is compound 76 Compound 86 or compound 87 5 . The peptoid polymer or a salt thereof according to any one of claims 1 to 3 , wherein the peptoid polymer forms a helical structure.

6. The peptoid polymer or a salt thereof according to any one of claims 1 to 3, wherein the salt of the peptoid polymer is selected from hydrochloride, acetate, sulfate, phosphate, maleate, citrate, methanesulfonate, nitrate, tartrate, gluconate or a combination thereof.

7. A peptoid-peptide hybrid or a salt thereof, comprising a peptoid polymer or a salt thereof according to any one of claims 1 to 6 and one or more amino acids, wherein the one or more amino acids are located at one or both ends of the peptoid polymer and / or between one or more subunits.

8. The peptoid-peptide hybrid or a salt thereof according to claim 7, wherein the one or more amino acids are selected from alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, arginine, lysine, leucine, methionine, asparagine, proline, glutamine, serine, threonine, valine, tryptophan, tyrosine or a combination thereof.

9. The peptoid-peptide hybrid or a salt thereof according to claim 8, wherein the one or more amino acids are selected from isoleucine, leucine, serine, threonine, alanine, valine, arginine or a combination thereof.

10. The peptoid-peptide hybrid or a salt thereof according to any one of claims 7 to 9, wherein the salt of the peptoid-peptide hybrid is selected from hydrochloride, acetate, sulfate, phosphate, maleate, citrate, methanesulfonate, nitrate, tartrate, gluconate or a combination thereof.

11. A cryopreservation solution comprising the peptoid polymer or a salt thereof according to any one of claims 1 to 6 or the peptoid-peptide hybrid or a salt thereof according to any one of claims 7 to 10, or a combination of the peptoid polymer or a salt thereof according to any one of claims 1 to 6 and the peptoid-peptide hybrid or a salt thereof according to any one of claims 7 to 10.

12. The cryopreservation solution according to claim 11, wherein the cryopreservation solution reduces or inhibits ice crystal formation at a temperature of 0°C to -20°C.

13. The cryopreservation solution according to claim 11, wherein the cryopreservation solution reduces or inhibits ice crystal formation at a temperature of -20°C to -40°C.

14. The cryopreservation solution according to claim 11, wherein the cryopreservation solution reduces or inhibits ice crystal formation at a temperature of -20°C.

15. The cryopreservation solution according to claim 11, wherein the cryopreservation solution reduces or inhibits ice crystal formation at a temperature of -40°C to -200°C.

16. A method for preserving tissues, organs or cells, the method comprising contacting the tissues, organs or cells with a peptoid polymer or a salt thereof according to any one of claims 1 to 6, a peptoid-peptide hybrid or a salt thereof according to any one of claims 7 to 10, a cryopreservation solution according to any one of claims 11 to 15, or a combination thereof.

17. The method of claim 16, wherein the peptoid polymer or salt thereof, peptoid-peptide hybrid or salt thereof, cryopreservation solution, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature of 0°C to -20°C.

18. The method of claim 16, wherein the peptoid polymer or salt thereof, peptoid-peptide hybrid or salt thereof, cryopreservation solution, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature of -20°C to -40°C.

19. The method of claim 16, wherein the peptoid polymer or salt thereof, peptoid-peptide hybrid or salt thereof, cryopreservation solution, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature of -20°C.

20. The method of claim 16, wherein the peptoid polymer or salt thereof, peptoid-peptide hybrid or salt thereof, cryopreservation solution, or a combination thereof is present in an amount sufficient to reduce or inhibit ice crystal formation at a temperature of -40°C to -200°C.

21. The method of any one of claims 16 to 20, wherein the tissue is a bioengineered tissue.

22. The method of any one of claims 16 to 20, wherein the peptoid polymer or salt thereof or the peptoid-peptide hybrid or salt thereof is present in an amount between 100 nM and 1,000 mM, or the combination of the peptoid polymer or salt thereof and the peptoid-peptide hybrid or salt thereof is present in an amount between 100 nM and 1,000 mM.

23. The method of any one of claims 16 to 20, wherein the tissue, organ or cell is selected from the group consisting of heart, liver, lung, kidney, pancreas, intestine, thymus, cornea, nerve cells, platelets, sperm cells, oocytes, corpus cavernosum cells, bladder cells, urethral cells, ureteral cells, testicular cells, embryonic cells, stem cells, immune system cells, bone cells, organoids or a combination thereof.

24. The method of claim 23, wherein the cells are primary cells.

25. The method of claim 23, wherein the stem cells are human pluripotent stem cells, hematopoietic stem cells, or a combination thereof.

26. The method of claim 23, wherein the immune system cell is a lymphocyte, a granulocyte, or a combination thereof.

27. A method for preserving a biomacromolecule, the method comprising contacting the biomacromolecule with a peptoid polymer or a salt thereof according to any one of claims 1 to 6, a peptoid-peptide hybrid or a salt thereof according to any one of claims 7 to 10, a cryopreservation solution according to any one of claims 11 to 15, or a combination thereof.

28. The method of claim 27, wherein the method comprises preserving DNA and RNA samples, stem cell growth factors or antibodies.

29. The method of claim 27, wherein the biomacromolecule is selected from nucleic acids, amino acids, proteins, peptides, lipids, or combinations thereof.

30. The method of claim 29, wherein the protein is an isolated protein.

31. The method of claim 30, wherein the isolated protein is a protease protein.

32. The method of claim 27, wherein the biomacromolecule is a complex structure.

33. The method of claim 32, wherein the composite structure is a liposome.

34. The method of claim 29, wherein the nucleic acid is DNA or RNA.

35. A beauty care product comprising the peptoid polymer or a salt thereof according to any one of claims 1 to 6, the peptoid-peptide hybrid or a salt thereof according to any one of claims 7 to 10, the cryopreservation solution according to any one of claims 11 to 15, or a combination thereof.

36. An antifreeze product comprising the peptoid polymer or a salt thereof according to any one of claims 1 to 6, the peptoid-peptide hybrid or a salt thereof according to any one of claims 7 to 10, the cryopreservation solution according to any one of claims 11 to 15 or a combination thereof.

37. An anti-icing product according to claim 36, wherein the anti-icing product is a de-icing or ice suppression product for preventing, inhibiting or delaying the formation of ice on or within an object.

38. The deicing product of claim 37, wherein the object is selected from an aircraft or part thereof, a gas pipeline, a window, an electrical device or a drone.

39. The antifreeze product of claim 38, wherein the electrical device is an electrical cable or power line.

40. The antifreeze product of claim 37, wherein the object is a mechanical device.

41. The antifreeze product of claim 40, wherein the mechanical device is an automobile engine, gear system or brake system.

42. A frozen food comprising the peptoid polymer or a salt thereof according to any one of claims 1 to 6, the peptoid-peptide hybrid or a salt thereof according to any one of claims 7 to 10, the cryopreservation solution according to any one of claims 11 to 15, or a combination thereof.

43. The frozen food of claim 42, wherein the frozen food is selected from ice cream, yogurt, seafood, fruit or meat products.

44. Use of the peptoid polymer or salt thereof according to any one of claims 1 to 6, the peptoid-peptide hybrid or salt thereof according to any one of claims 7 to 10, the cryopreservation solution according to any one of claims 11 to 15, or a combination thereof in the following aspects: - For the preservation of tissues, organs or cells; - Use for preservation of biomacromolecules; - Use in the preparation of beauty care products; - Use in the preparation of an antifreeze product; or - Use in the preparation of frozen foods.

45. Use according to claim 44, wherein the anti-icing product is a de-icing or ice suppression product for preventing, inhibiting or delaying the formation of ice on or in objects.

46. ​​Use according to claim 45, wherein the object is selected from an aircraft or a part thereof, a gas pipeline, a window, an electrical device or a drone.

47. Use according to claim 46, wherein the electrical device is a cable or a power line.

48. The use according to claim 45, wherein the object is a mechanical device.

49. The use according to claim 48, wherein the mechanical device is an automobile engine, a gear system or a brake system.

50. The use according to claim 44, wherein the frozen food is selected from ice cream, yogurt, seafood, fruit or meat products.

Images