WO2021252607A1

WO2021252607A1 - Beta-propiolactone compounds and ring opened beta-propiolactone compounds and uses for same

Info

Publication number: WO2021252607A1
Application number: PCT/US2021/036581
Authority: WO
Inventors: Eric STOUTENBURG; Jeff UHRIG; Geoff Coates
Original assignee: Novomer, Inc.
Priority date: 2020-06-12
Filing date: 2021-06-09
Publication date: 2021-12-16

Abstract

Functionalized beta-propiolactone in which one or more of the carbons or the ring structure are substituted with either hydrogen or a hydrocarbyl moiety which may contain heteroatoms and substituents.

Description

BETA-PROPIQLACTONE COMPOUNDS AND RING OPENED

BETA-PROPIOLACTONE COMPOUNDS AND USES FOR SAME

BACKGROUND

[0001] The present invention is a non-provisional utility application that claims priority to Unites States Provisional Patent Application Serial Number 63/038,533 filed June 12, 2020, currently pending.

[0002] The present disclosure is directed to the use of beta-propiolactone compounds, particularly functionalized beta-propiolactone compounds in the ring-opened and ring-closed state. This disclosure is also directed to uses for functionalized beta-propiolactone compounds for reactions biologically derived material such as material including nucleic acid such uses including, but not limited to, use as an active agent against biological pathogens during vaccine preparation as well as the processing of biological specimens and materials.

[0003] Various pharmaceutical, biomedical and bioanalytical processes incorporate reactions with biologically derived material reactions that include one or more macromolecular biopolymers. These can include stabilizing blood for transport, inactivation of biological pathogens such as viral material and various analytical procedures. Non-limiting examples of such processes include purification of biological material such as blood products and biological specimens, analysis and/or preservation of biological specimens as well as processes associated with preparation of vaccines.

[0004] Vaccine production can involve inactivation of the bacterial or viral material.

Bacterial or viral inactivation can include cross linking the nucleic acid present in the bacterial or viral material. Compounds that can react with nucleic acid without undo reaction of interaction with various other biological compounds present would be valuable for the use in biological purification processes, analysis and/or preservation of biological specimens or vaccine preparation.

[0005] The present disclosure provides methods and compounds that can modify the function of biologically derived material such as those containing macromolecular biopolymer. The macromolecular biopolymer can be selected from the group consisting of nucleic acids, proteins, and carbohydrates. The macromolecular biopolymer in question can be present in biological pathogens such as bacteria, viruses, and the like. More particularly, the present disclosure provides methods and compounds that can be employed in processes and procedures that require full or partial chemical inactivation of viral material while preserving functional viral proteins that enhance antibody production and future pathogen recognition in the target individual.

SUMMARY

[0006] Disclosed is functionalized beta-propiolactone having the general formula:

wherein R¹, R², R³ , R⁴ are hydrogen or a hydrocarbyl moiety which may contain heteroatoms and substituents.

[0007] Also disclosed is a ring-opened functionalized beta-propiolactone complex derived from the structure corresponding to the general formula set forth above. The ring-opened functionalized beta-propiolactone complex can have the formula:

in which R¹ and R² are hydrogen or a hydrocarbyl moiety which may contain heteroatoms and substituents at each occurrence are independently with at least one R¹ and R² being a hydrocarbyl moiety. A denotes biologically derived material.

[0008] The hydrocarbyl moiety R¹ , R² present in the ring-opened functionalized beta- propiolactone compound is one that enhances the function of the ring-opened functionalized beta-propiolactone compound in contact with the biologically derived material. [0009] In certain embodiments, the biologically derived material A can be a biological pathogen such as bacterial, viruses and the like. In certain embodiments, the virus can be a virus from the family Picornaviridae or Coronaviridae.

[0010] Also disclosed is a method for producing functionalized beta-propiolactone that includes the steps of introducing a beta-propiolactone feed stock having elevated purity into contact with a strong base in a polar aprotic solvent at a temperature below -50C and introducing a reactive adduct into contact with the beta-propiolactone feed stock. The reactive adduct includes a moiety which, when present on the beta-propiolactone molecule, imparts enhances the capability of the synthesized compounds to function that cause cross linkage of a biologically derived chemical compound.

DETAILED DESCRIPTION

[0011] The present disclosure is directed to functionalized beta-propiolactone compounds that can react with biologically derived material in which the function of the biologically derived material is altered. The functionalized beta-propiolactone compounds disclosed can react with at least one chemical site present at least one compound in the biologically derived material. In certain embodiments, the functionalized beta-propiolactone compounds as disclosed can include at least one functional group that facilitates interaction with a macromolecular biopolymer such as nucleic acids, proteins, and carbohydrates. In certain embodiments, the macromolecular biopolymer can be a nucleic acid or nucleic acid fragment. In certain embodiments the nucleic acid or nucleic acid fragment can contain a substituted purine reaction site. It is believed that the functionalized beta-propiolactone as disclosed may interact with macromolecular biopolymers such as nucleic acids or nucleic acid fragments present in biological pathogens to modify the function of the macromolecular biopolymer. The biological pathogen may include bacteria, viruses, and the like in certain embodiments.

[0012] Also disclosed are functionalized beta-propiolactone compounds that can complex with biologically derived material as well as complexes of the functionalized beta- propiolactone compound(s) pathogens such as viruses and/or bacteria in a manner that modifies the function of the biological pathogen as well as complexes of the beta-functionalized beta- propiolactone and biologically derived material. [0013] The functionalized beta-propiolactone compound can be employed in various processes and methods including, but not limited to purification of biomaterials, bioanalytical methods, vaccine preparation and the like. The functionalized beta-propiolactone compound disclosed can complex with and inactivate biological pathogens while maintaining biological integrity of specific proteins such as those capable of eliciting and/or initiating an antibody response when introduced into an individual.

[0014] The beta-propiolactone compound useful for inactivating the target virus can be functionalized with one of more organic moieties that are capable of enhancing the function of the disclosed compounds to function as chemical inactivation agents. The hydrocarbyl moieties may impart sufficient lipophilicity to the functionalized beta-propiolactone compound to permit its transit through a lipid layer present in a virus structure.

[0015] Where desired, the functionalized beta-propiolactone compounds can be prepared from beta-propiolactone processes that yield beta-propiolactone product having a purity above 90%, in some embodiments, the beta-propiolactone compounds can have a purity greater than 95%, greater than 98% and in some embodiments above 99%.

[0016] The functionalized beta-propiolactone as disclosed can be one that is substantially polymer-free. In certain embodiments, the beta-propiolactone that is employed can have aforementioned purity and may contain a suitable stabilizer such as cyclic anhydrides and the like.

[0017] The beta-propiolactone compound as disclosed can have the general formula:

wherein R¹, R², R³ , R⁴ are hydrogen or a hydrocarbyl moiety which may contain heteroatoms and substituents. R¹, R², R³, R⁴ can be present as a hydrocarbyl moiety that enhances the function of the beat-propiolactone in contact with a biologically derived material.

In certain embodiments, each of R¹, R², R³, R⁴ can be selected to modify steric hindrance of the beta-propiolactone molecule, modify electron density of the beta-propiolactone molecule, modify the lipophilicity of the beta-propiolactone molecule and the like. [0018] The biologically derived material can be one that includes at least one macromolecular biopolymer. The at least one macromolecular biopolymer can be selected from the group consisting of nucleic acids, proteins, and carbohydrates. In situations where the macromolecular biopolymer is one or more nucleic acid, the nucleic acid can be present in suitable biological structures such in cells or subcellular material. The biologically derived material can be biological pathogens such as bacteria, viruses, and the like. Non-limiting examples of viruses include those from the family Picornaviridae or the family Coronaviridae. The biologically derived material can be the nucleic acid that is present in or associated with the biological pathogen. Where desired or required, the nucleic acid can have a reaction site that includes at least one substituted purine constituent and the functionalized beta-propiolactone interacts with at least that substituted purine constituent site in a manner that modifies the associated biologically derived material.

[0019] It is believed that the functionalized beta-propiolactone compound as disclosed can, in certain embodiments, react with biologically derived material such that containing nucleic acid to cross link the nucleic acid at one or more locations thereby altering the function of some or all of the nucleic acid. The nucleic acid can be a single stranded nucleic acid or a double stranded material such as ribonucleic acid or deoxyribonucleic acid. Without being bound to any theory, it is believed that the functionalized beta-propiolactone can interact with nitrogenous bases present in the nucleic acid. In certain embodiments, the nitrogenous base targeted can be one or more substituted purine groups as would be found in adenine, guanine and/or one or more pyrimidine groups as would be found in cytosine, thymine, or uracil.

[0020] R¹, R², R³, R⁴ may be separately in each occurrence an aryl, substituted aryl, cycloaliphatic, substituted cycloaliphatic, aliphatic or substituted aliphatic moiety. Each may be alcohol, carboxylic acid, dicarboxylic acid, substituted indole, substituted indene, triglyceride moieties and the like

[0021] Each of R¹, R², R³, R⁴ may be separately in each occurrence alkyl, alkenyl, cycloalkyl, heterocyclyl, alkyl heterocyclyl, aryl, aralkyl, alkaryl, heteroaryl, or alkyl heteroaryl, or polyoxyalkylene, or two of R¹, R², R³, R⁴ may form a 5-7 membered cyclic or heterocyclic ring. R¹, R², R³, R⁴ may be separately in each occurrence C1--C15 alkyl, C2-C15 alkenyl, C3-C9 cycloalkyl, C2-C20 heterocyclyl, C3-C20 alkyl heterocyclyl, C6-Cis aryl, C7-C25 alkaryl, C7-C25 aralkyl, Cs-Cis heteroaryl or C6-C₂₅ alkyl heteroaryl, or polyoxyalkylene. The recited groups may be substituted with one or more substituents, which do not interfere with the uses of these compounds as disclosed herein. Exemplary substituents include halo, alkylthio, alkoxy, hydroxyl, nitro, azido, cyano, acyloxy, carboxy, ester or unsaturated groups two of R¹, R², R³ , R⁴ may be separately in each occurrence C₁-C₁₅ alkyl, C3-C6 cycloalkyl, C₄-C₁₈ heterocyclyl, C₄- Ci8 alkheterocyclyl, C6-C₁₈ aryl, C7-C₂₅ alkaryl, C7-C₂₅ aralkyl, C5-C₁₈ heteroaryl or C6-C₂₅ alkyl heteroaryl. Heteroatom as used herein means oxygen, nitrogen, sulfur, and phosphorus.

[0022] The functionalized beta-propiolactone compound as disclosed can have at least one of R¹, R², R³, R⁴ present as a hydrophilic lipid selected from the group consisting of substituted free fatty acids, unsubstituted free fatty acids, substituted triglycerides, unsubstituted triglycerides, substituted phospholipids, unsubstituted phospholipids, substituted polyketids, unsubstituted poly ketids, substituted phosphoglycerides, unsubstituted phosphoglycerides. The degree of lipophilicity imparted can be that sufficient to permit transit through a lipid layer present in the biological material such as in a virus structure. In certain embodiments, R¹, R², R³, R⁴ may be selected to provide steric hindrance modification to the molecule and/or to accomplish molecular electron density distribution in the molecule. It is contemplated that the functionalized beta-propiolactone can include a combination of functional groups that provide one or more desired modifications.

[0023] In certain embodiments, R¹, R², R³, R⁴ can each be a hydrocarbyl moiety that is the same or different from the other hydrocarbyl moieties. In certain embodiments at least three of R¹, R², R³, R⁴ can be a respective hydrocarbyl moiety. In certain embodiments at least two or one of R¹, R², R³, R⁴ can be a respective hydrocarbyl moiety.

[0024] The functionalized beta-propiolactone compound of contacting a biologically derived material and may complex with the desired target material such as a biological pathogen. In certain embodiments, the biological pathogen can be one or more bacterial or viral materials present in biological samples such as blood products which need to be inactivated, neutralized or eliminated. In other embodiments, the biological pathogen can be a target virus to be inactivated in the process of vaccine preparation. In vaccine formation the virus of interest can be inactivated or partially inactivated as by inactivating nucleic acid present in the biological pathogen. [0025] The virus of interest can be one having a structure that includes nucleic acid that can be inactivated or partially inactivated by interaction with the functionalized beta- propiolactone compound in either its ring-open or ring-closed structure. The virus can be non- enveloped or can have a viral envelope. Where present, the viral envelope can be a mixture of phospholipids, proteins, as well as glycoproteins derived from viral material. It is believed that particular stmctural and biochemical features present on the exterior surface of the virus such as the viral envelope can serve as antigens that can facilitate antibody response to the presence of virus in the body of the patient.

[0026] Non-limiting examples of viruses that can be treated by the beta-propiolactone compound as disclosed can include viruses from the family Picornaviridae or the family Coronaviridae. Non-limiting examples of picornaviruses include enteroviruses, saphthoviruses, cardioviruses, hepatoviruses. Specific viruses include picornaviruses include poliovirus, coxsackievims and the like. Non-limiting examples of coronaviruses include betacoronavirus such as severe acute respiratory syndrome related coronavirus such as SARS-CoV, SARS-CoV- 2, Middle east respiratory related coronavirus such as MERS-CoV.

[0027] The functionalized beta-propiolactone compound as disclosed can be synthesized by a method in which a beta-propiolactone feed stock having a purity greater than 98%. In certain embodiments, a beta-propiolactone feed stock having a purity greater than 99% can be used. High purity beta-propiolactone feed stock can be prepared by reacting a feed stream containing an epoxide and carbon monoxide in the presence of a carbonylation catalyst to produce a beta-lactone stream comprising beta-propiolactone. The epoxide component can be either substituted or unsubstituted. Where desired or required, the high purity beta-propiolactone feed stock can be prepared according to the disclosure found in U.S. Patent Nos. 8,445,703; 8,796,475; 9,206,144 and 9,493,391, the specifications of which are incorporated by reference herein.

[0028] One suitable carbonylation catalyst comprises a metal carbonyl compound balanced by a suitable Lewis acid in which the metal carbonyl compound has the general formula [QM_y(CO)_w]_x where Q is any ligand and need not be present; M is a metal atom; y is an integer from 1 to 6 inclusive; w is a number that provides a metal carbonyl compound that is stable, and x is an integer from -3 to +3 inclusive. In certain embodiments, M can be Ti, Cr, Mn, Fe, Ru, Co, Rh, Ni, Pd, Cu, Zn, Al, Ga and In. In certain embodiments, M can be cobalt. In certain embodiments, y is an integer between 1 and 3. In certain embodiments, y is 1. In certain embodiments, w is 4. In certain embodiments, the metal carbonyl compound comprises a carbonyl cobaltate and the carbonylation catalyst further comprises a Lewis acidic co-catalyst which comprises a metal-centered cationic Lewis acid comprising an aluminum cation.

[0029] In certain embodiments, the carbonylation catalyst further comprises a Lewis acidic component. In some embodiments, the carbonylation catalyst includes an anionic metal carbonyl complex (e.g. x is a negative integer) and a cationic Lewis acidic component. In certain embodiments, the metal carbonyl complex comprises a carbonyl cobaltate and the Lewis acidic co-catalyst comprises a metal-centered cationic Lewis acid.

[0030] In certain embodiments, the metal-centered Lewis acid is a metal complex of formula [M’(L)_¾]^C+, where: M’ is a metal; each L is a ligand; b is an integer from 1 to 6 inclusive; c is 1, 2, or 3; and where, if more than one L is present, each L may be the same or different.

In some embodiments where the metal-centered Lewis acid is a metal complex of formula [M’(L)_&]^C+, M’ is selected from the group consisting of: a transition metal, a group 13 or 14 metal, and a lanthanide. In certain embodiments, M’ is a transition metal or a group 13 metal. In certain embodiments, is selected from the group consisting of aluminum, chromium, indium and gallium. In certain embodiments, M’ is aluminum. In certain embodiments, M’ is chromium. [0031] Various suitable carbonylation catalysts discussed in and synthetic processes are described in U.S. Patent Numbers 10,221,150 and 10,590,099 and U.S. Published Patent Application 2018-0029005 the specifications of which are each incorporated by reference herein. [0032] In the synthetic process as disclosed, an epoxide compound, a solvent, a carbonylation catalyst, and carbon monoxide can be contacted inside a reactor to produce a first product stream comprising beta-propiolactones and the carbonylation catalyst. Upon completion of the reaction process, the first product stream is separated to produce the carbonylation catalyst and a beta-propiolactone stream comprising the beta-propiolactones having a purity of about 99 percent or more.

[0033] The solvent that is employed in the synthesis process can be a solvent having a boiling point higher than about 108 at 50 Torr since the lactone produced (propiolactone) has a boiling point of 108 °C at that pressure. For propylene oxide, the corresponding lactone (beta- butyrolactone) has a boiling point of 86-87 °C at 50 Torr, and the high boiling solvent for these processes should have a higher boiling point than that. In certain embodiments, the boiling point of the high boiling solvent is selected to be at least 20 degrees higher than that of the lactone produced by the carbonylation step. In certain embodiments, the boiling point of the high boiling solvent is selected to be at least 30 degrees higher than that of the lactone produced by the carbonylation step. In certain embodiments, the boiling point of the high boiling solvent is selected to be at least 50 degrees higher than that of the lactone produced by the carbonylation step. In certain embodiments, the high boiling solvent is selected from the group consisting of: sulfolane; /V- methyl pyrrolidone; l,3-dimethyl-2-imidazolidinone; diglyme; triglyme; tetraglyme; diethylene glycol dibutyl ether; ethylene carbonate; propylene carbonate; butylene carbonate; dibasic esters; and mixtures of any two or more of these. In general, polar aprotic solvents, or hydrocarbons are suitable for this step. In certain embodiments, protic solvents are unsuitable for the first step. Where desired or required, the solvents as well as all feed stocks and intermediates can be substantially free of formaldehyde, formaldehyde derivatives and formaldehyde-containing compounds as well as compounds that can be devolved to the same during processing and manufacture. The resulting beta-propiolactone material that is synthesized can be non-functionalized or partially functionalized.

[0034] Where desired or required, the resulting beta-propiolactone material functionalized by any suitable process. Non-limiting examples of suitable synthetic processes include For example, the resulting beta-propiolactone material can be brought into contact with a strong base in a polar aprotic solvent into which a reactive compound X-R is introduced. In the reactive compound X-R, X can be a suitable leaving group including but not limited to halogens, tosylates, mesylates, and the like. R can be a hydrocarbyl group as defined previously and may be selected from the group including but not limited to an aryl, substituted aryl, cycloaliphatic, substituted cycloaliphatic, aliphatic or substituted aliphatic moiety.

[0035] Also disclosed is a ring-opened functionalized beta-propiolactone constituent having the general formula:

in which R¹ and R² is each independently hydrogen or a hydrocarbyl moiety. The hydrocarbyl moiety can optionally contain at least one heteroatom or at least one substituent in which at least one of R¹, R² are present a hydrocaryl moiety and wherein

indicates attachment to a suitable biological structure or intermediate functional group. Where R¹ or R² is a hydrocarbyl moiety that enhances the function of the disclosed compounds to function relative to an associated biologically active material substituted alkyl or substituted aryl, R¹ and/ or R² will include at least one functional moiety as defined above.

[0036] The ring-opened functionalized beta-propiolactone complex derived from the structure of formula corresponding to the general formula of the functionalized beta- propiolactone set forth above. One embodiment of the ring-opened functionalized beta- propiolactone complex can have the formula:

in which R¹ and R² are hydrogen or a hydrocarbyl moiety which may contain heteroatoms and substituents, wherein at least one of R¹ and R² is the hydrocarbyl moiety that enhances the function of the disclosed compounds to function as chemical inactivation agents. R¹ and R² may be separately in each occurrence an aryl, substituted aryl, cycloaliphatic, substituted cycloaliphatic, aliphatic or substituted aliphatic moiety. Each R¹ and R² may be alcohol, carboxylic acid, dicarboxylic acid, substituted indole, substituted indene, triglyceride moieties and the like.

[0037] Each of R¹ and R² may be separately in each occurrence alkyl, alkenyl, cycloalkyl, heterocyclyl, alkyl heterocyclyl, aryl, aralkyl, alkaryl, heteroaryl, or alkyl heteroaryl, or polyoxyalkylene, or R¹ and R² may form a 5-7 membered cyclic or heterocyclic ring. R¹ and R² may be separately in each occurrence CTC15 alkyl, C2-C15 alkenyl, C3-C9 cycloalkyl, C2-C20 heterocyclyl, C3-C20 alkyl heterocyclyl, C6-C18 aryl, C7-C25 alkaryl, C7-C25 aralkyl, C5-C18 heteroaryl or C6-C25 alkyl heteroaryl, or polyoxyalkylene, or R'and R² may form groups form a 5-20 membered cyclic or heterocyclic ring. The recited groups may be substituted with one or more substituents, which do not interfere with the uses of these compounds as disclosed herein. Exemplary substituents include halo, alkylthio, alkoxy, hydroxyl, nitro, azido, cyano, acyloxy, carboxy, ester or unsaturated groups. R¹ and R² may be separately in each occurrence C1-C15 alkyl, C3-C6 cycloalkyl, C4-C18 heterocyclyl, C4-C18 alkheterocyclyl, C6-C18 aryl, C7-C25 alkaryl, C7-C25 aralkyl, Cs-Cis heteroaryl or C6-C25 alkyl heteroaryl. Heteroatom as used herein means oxygen, nitrogen, sulfur and phosphorus.

[0038] Each of R¹ and R² may be separately in each occurrence alkyl, alkenyl, cycloalkyl, heterocyclyl, alkyl heterocyclyl, aryl, aralkyl, alkaryl, heteroaryl, or alkyl heteroaryl, or polyoxyalkylene, or R¹ and R² may form a 5-7 membered cyclic or heterocyclic ring. R¹ and R² may be separately in each occurrence CTC15 alkyl, C2-C15 alkenyl, C3-C9 cycloalkyl, C2-C20 heterocyclyl, C3-C20 alkyl heterocyclyl, C6-C18 aryl, C7-C25 alkaryl, C7-C25 aralkyl, C5-C18 heteroaryl or C6-C25 alkyl heteroaryl, or polyoxyalkylene, or R'and R² may form groups form a 5-20membered cyclic or heterocyclic ring. The recited groups may be substituted with one or more substituents, which do not interfere with the uses of these compounds as disclosed herein. Exemplary substituents include halo, alkylthio, alkoxy, hydroxyl, nitro, azido, cyano, acyloxy, carboxy, ester or unsaturated groups. R¹ and R² may be separately in each occurrence C1-C15 alkyl, C3-C6 cycloalkyl, C4-C18 heterocyclyl, C4-C18 alkheterocyclyl, C6-C18 aryl, C7-C25 alkaryl, C7-C25 aralkyl, C5-C18 heteroaryl or C6-C25 alkyl heteroaryl. Heteroatom as used herein means oxygen, nitrogen, sulfur and phosphorus.

[0039] A is an attached group. The attached group can be a biological pathogen. The biological pathogen can be at least one strand or strand fragment of nucleic acid. Biological pathogens can include bacteria, viruses or fragments of the same. In certain embodiments, the virus structure can be a virus from the family Picornaviridae or Coronaviridae.

[0040] The present disclosure also contemplates methods for inactivating a virus that include the steps of contacting the virus material with the resulting beta-propiolactone compound for an interval sufficient to produce an organo-viral complex in which the virus in contact with one or more alpha carbons of the beta-propiolactone compound, one or more beta carbons of the beta-propiolactone compound, or any combination thereof.

[0041] The resulting functionalized beta-propiolactone compounds can be employed to inactivate various viruses. It has been found that the resulting beta-propiolactone compounds can inactivate the desired vims without unduly compromising proteins necessary for achieving antibody response in the associated patient. It has been posited that beta-propiolactone compounds can affect cross linking one or more amine constituents present in the viral nucleic acid.

[0042] As broadly construed, reactive contact between the desired functionalized beta- propiolactone compound and the vims can produce an organo-viral complex having at least one of the following general formulae:

in which each R¹, R² are hydrogen or hydrocarbyl groups in which a given hydrocarbyl moiety may contain heteroatoms and substituents and wherein at least one of R¹, R² is a hydrocarbyl moiety that enhances the function of the disclosed compounds to function as chemical inactivation agents. The value n can be one or a number greater than one.

[0043] It is contemplated that the number of ring-opened functionalized beta- propiolactone moieties associated with a given virus structure will be that sufficient to accomplish inactivation of the nucleic acid present in the virus. It is contemplated that the organo-viral complex as disclosed herein can include compounds in which the ratio of individual virus to functionalized beta-propiolactone compound can be at least 1:1. It is contemplated that the ratio of virus to functionalized beta-propiolactone groups can be in a range between 1 to 1 and the number of reactive sites present in the virus. These reactive sites may include binding sites located in one or more virus structures such as on the surface of the capsid.

[0044] Where R ¹ and/or R² is a hydrocarbyl moiety, each of R¹, R² separately can be an aryl, substituted aryl, cycloaliphatic, substituted cycloaliphatic, aliphatic or substituted aliphatic moiety. Each of R¹, R² separately can be selected from alkyls, alkoxyls, carboxylic acids, dicarboxylic acids, substituted indoles, substituted indenes, triglycerides. Each of R¹, R² can be chosen to enhance to function of the disclosed compounds to function as chemical inactivation agents as by enhancing the lipophilicity of the compound, modifying steric hindrance characteristics of the compound, modifying the electron density within the compound and the like.

[0045] Each R¹, R² can be substituted or unsubstituted. Each of R¹, R² may be separately in each occurrence alkyl, alkenyl, cycloalkyl, heterocyclyl, alkyl heterocyclyl, aryl, aralkyl, alkaryl, heteroaryl, or alkyl heteroaryl, or polyoxyalkylene, or the two of R¹, R² may form a 5-7 membered cyclic or heterocyclic ring. R¹, R² may be separately in each occurrence Ci-Ci₅ alkyl, C₂-C₁₅ alkenyl, C₃-C₉ cycloalkyl, C₂-C₂₀ heterocyclyl, C₃-C₂₀ alkyl heterocyclyl, C6-C18 aryl, C7-C25 alkaryl, C7-C25 aralkyl, C5-C18 heteroaryl or C6-C25 alkyl heteroaryl, or polyoxyalkylene, or the two of R¹, R² may form groups form a 5-20 membered cyclic or heterocyclic ring. The recited groups may be substituted with one or more substituents, which do not interfere with the uses of these compounds as disclosed herein. Exemplary substituents include halo, alkylthio, alkoxy, hydroxyl, nitro, azido, cyano, acyloxy, carboxy, ester or unsaturated groups. The two of R¹, R² may be separately in each occurrence C₁-C₁₅ alkyl, C3-C6 cycloalkyl, C₄-C₁₈ heterocyclyl, C₄-C₁₈ alkheterocyclyl, C6-Cis aryl, C₇-C₂₅ alkaryl, C₇-C₂₅ aralkyl, Cs-Cis heteroaryl or C6-C₂₅ alkyl heteroaryl. Heteroatom as used herein means oxygen, nitrogen, sulfur and phosphorus.

[0046] Substituent groups, where present can be selected from the group consisting of substituted free fatty acids, unsubstituted free fatty acids, substituted triglycerides, unsubstituted triglycerides, substituted phospholipids, unsubstituted phospholipids, substituted polyketids, unsubstituted poly ketids, substituted phosphoglycerides, unsubstituted phosphoglycerides, alkyls, alcohols carboxylic acids, dicarboxylic acids, substituted indoles, substituted indenes, and the like.

[0047] In the organo- viral complex as depicted, “virus” is understood to be an infective agent that includes at least one strand of nucleic acid. The nucleic acid can be surrounded or enveloped by a structural layer that is composed of various proteins, lipids, phospolipids and the like. [0048] In certain embodiments, the vims is from the family Picornaviridae or

Coronaviridae and is one of coronavirus, enterovirus, aphthovims, cardiovirus, hepatovims, and the like. In certain embodiments, the vims is one of the following: coronavirus, coxsackievirus, rhinovirus. In certain embodiments, the virus can be one of SARS-CoV, SARS-CoV-2, MERS- CoV.

[0049] The connection between the vims and the ring-opened functionalized beta- propiolactone moieties can be by any suitable chemical bond. In certain embodiments, the bond can be a suitable covalent bond. Bonding between the ring-opened functionalized beta- propiolactone and the virus can involve proteins present on any suitable viral stmcture of the vims. Non-limiting examples of such proteins include nucleoproteins, membrane proteins, spike glycoproteins, envelope proteins, and small membrane protein. It is believed that ring-opened functionalized beta-propiolactone moieties may have affinity to cysteine, methionine, and/or histidine in certain embodiments.

[0050] Without being bound to any theory, it is hypothesized that the presence of functional groups on the functionalized beta-propiolactone compound can render the ring opened compound more amenable to reaction with adenine and/or guanine present in nucleic acid in the viral structure. It is believed that the presence of functionalized groups can modulate the ring opening kinetics as well as reactivity of the compound with associated material.

[0051] It is believed that the functionalized beta-propiolactone can interact with the adenine portion of a nucleic acid to achieve crosslinking of the associated nucleic acid. One theoretic method is outlined below in which the adenine ring opens the propionic ring of a beta propionic acid and achieves deactivation of RNA through alkylation of the adenine nucleotide.

[0052] The beta-propiolactone include one or more functional groups. An illustrative synthetic method in which a functional group is a triglyceride is depicted. The resulting triglyceride epoxide can be a regioisomeric mixture. Various triglycerides can undergo suitable reactions to yield triglyceride epoxides. The resulting triglyceride epoxides can be a regioisomeric mixture. The resulting triglyceride epoxide can undergo a suitable beta- propiolactone formation reaction as disclosed herein to yield a functionalized beta-propiolactone triglyceride :

[0053] The functionalized beta-propiolactone can be synthesized by any suitable manner such as a suitable reaction. In certain processes, the suitable functionalized beta-propiolactone can be prepared using a halogen compound that includes a good leaving group in the presence of a strong base in a protic solvent. A non-limiting reaction scheme is depicted below:

A

The reaction can also be employed to impart aryl functionality as depicted below:

B [0001] Beta-propiolactone synthesis can proceed using any suitable method. The propiolactone can be synthesized by the catalytic method as outlined previously and depicted in Scheme C with the resulting compound being functionalized subsequently. It is also contemplated that beta- propiolactone can be synthesized as depicted in Scheme D. Functionalized beta-propiolactone can be synthesize by various methods such as those illustrated in Scheme E and F using the catalysts outlined.

O catalyst

C.

catalyst

D.

In each of the preceding reaction process diagrams R¹ and R² can be as described before and are independent of one another.

[0054] Once introduced into contact with the target virus to be inactivated, the functionalized beta-propiolactone compound as disclosed can be undergo a ring opening reaction, typically either by an acylation reaction, an alkylation reaction or both at the beta carbon to yield ring open structures. Where desired or required, one or more of the functional groups can be selected to direct or promote either the acylation reaction or the alkylation reaction.

[0055] The functionalized ring opened beta-propiolactone can be complexed to one more structural elements of the target virus. In certain embodiments, it is believed that the functionalized ring opened beta-propiolactone is connected to structures present in the virus through covalent bonding and can establish organo-viral complexes as outlined in the reaction schemes below:

in which R¹ and or R² are functional groups as discussed previously.

in which R¹ and or R² are functional groups as discussed previously.

[0056] The present disclosure also contemplates methods for inactivating a virus that include the steps of contacting the virus material with the resulting beta-propiolactone compound for an interval sufficient to produce an organo-viral complex in which the virus in contact with one or more alpha carbons of the beta-propiolactone compound, one or more beta carbons of the beta-propiolactone compound, or any combination thereof.

[0057] The resulting functionalized beta-propiolactone compounds can be employed to inactivate various viruses. It has been found that the resulting beta-propiolactone compounds can inactivate the desired vims without unduly compromising proteins necessary for achieving antibody response in the associated patient. It has been posited that beta-propiolactone compounds can affect cross linking one or more amine constituents present in the viral nucleic acid.

[0058] In certain embodiments the number n can be between 1 and 1000; with n between

1 and 100 in some embodiments.

[0059] In certain embodiments, the virus is from the family Picornaviridae or

Coronaviridae and is one of coronavirus, enterovirus, aphthovirus, cardiovirus, hepatovirus, and the like. In certain embodiments, the virus is one of the following: coronavirus, coxsackievirus, rhinovirus. In certain embodiments, the virus can be one of SARS-CoV, SARS-CoV-2, MERS- CoV.

[0060] Also disclosed is a method for inducing immune response in a human in which the method comprises administering to the human a composition comprising a chemically inactivated coronavirus that has been inactivated by treatment with a beta-propiolactone compound, the inactivated coronavirus is administered in an amount sufficient to induce an immune response in the patient.

[0061] In certain embodiments, it is believed that one or more beta-propiolactone compounds can covalently attach to peptides present in the coronavirus structure, as in areas of active peptides contained within the “spike” of the coronavirus. It is believed that reaction with the spike may enable deactivation of the virus while still allowing the body to generate antibodies to defend against similar peptide sequences. It is believed that the functionalized beta- propiolactone compounds are favored to pass through the envelop structure that is characterized by an oily layer. It is believed that the oleophilic functionalization as disclosed herein can assist in maintaining the integrity of some or all of the viral envelop structure while allowing the beta- lactone structure to ring opening the presence of thiols accessible in the cysteine contained in the coronavirus.

[0062] While the invention has been described in connection with what is presently considered to be the most practical and preferred embodiment, it is to be understood that the invention is not to be limited to the disclosed embodiments but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims, which scope is to be accorded the broadest interpretation so as to encompass all such modifications and equivalent structures as is permitted under the law.

Claims

What is claimed is:

1. A functionalized beta-propiolactone having the general formula:

wherein R¹, R², R³, R⁴ are hydrogen or a hydrocarbyl moiety, the hydrocarbyl moiety optionally containing at least one heteroatom or at least one substituent, wherein at least one R¹, R², R³, R⁴ are present as a hydrocarbyl.

2. The functionalized beta-propiolactone of claim 1 wherein at least at least one R¹, R², R³, R⁴ are present as a hydrocarbyl moiety that enhances function of the functionalized beta- propiolactones in contact with biologically derived material.

3. The functionalized beta-propiolactone of claim lor 2 wherein R¹, R², R³, R⁴ are separately and in each occurrence hydrogen or an aryl, substituted aryl, cycloaliphatic, substituted cycloaliphatic, aliphatic or substituted aliphatic moiety.

4. The functionalized beta-propiolactone of claims 1, 2, 3 or 4 wherein R¹, R², R³, R⁴ separately and at each occurrence are hydrogen, alkyl, alkenyl, cycloalkyl, heterocyclyl, alkyl heterocyclyl, aryl, aralkyl, alkaryl, heteroaryl, or alkyl heteroaryl, or polyoxyalkylene, or two of R¹, R², R³, R⁴may form a 5-20 membered cyclic or heterocyclic ring, R¹, R², R³, R⁴ may be separately in each occurrence C1-C15 alkyl, C2-C15 alkenyl, C3-C9 cycloalkyl, C2-C20 heterocyclyl, C3-C20 alkyl heterocyclyl, C6-Cis aryl, C7-C25 alkaryl, C7-C25 aralkyl, C5-C18 heteroaryl or C6-C25 alkyl heteroaryl, or polyoxyalkylene.

5. The functionalized beta-propiolactone of claims 1, 2, 3 or 4 wherein the substituent is halo, alkylthio, alkoxy, hydroxyl, nitro, azido, cyano, acyloxy, carboxy, ester or unsaturated groups two of R¹, R², R³, R⁴ may be separately in each occurrence C1-C15 alkyl, C3-C6 cycloalkyl, C4- Ci8 heterocyclyl, C4-C18 alkheterocyclyl, Ce-Cis aryl, C7-C25 alkaryl, C7-C25 aralkyl, C5-C18 heteroaryl or C6-C25 alkyl heteroaryl.

6. The functionalized beta-propiolactone of claim 1, 2, 3, 4 or 5 wherein R¹, R², R³, R⁴ is halogen or a hydrocarbyl selected from the group consisting of alkyls, alkoxyls carboxylic acids, dicarboxylic acids, substituted indoles, substituted indenes, triglycerides, and mixtures thereof.

7. The functionalized beta-propiolactone of claims 1, 2, 3, 4, 5 or 6 wherein the substituent is selected from the group consisting of substituted fatty acids, unsubstituted fatty acids, substituted triglycerides, unsubstituted triglycerides, substituted phospholipids, unsubstituted phospholipids, substituted polyketids, unsubstituted polyketids, substituted phosphoglycerides, unsubstituted phosphoglycerides and mixture thereof.

8. The functionalized beta-propiolactone of claims 1, 2, 3, 4, 5, 6 or 7 wherein the heteroatom is oxygen, nitrogen, sulfur and phosphorus.

9. The functionalized beta-propiolactone of claims 1, 2, 3, 4, 5, 6, 7 or 8 wherein the at least one R¹, R², R³, R⁴ are present as a hydrocarbyl moiety that enhances reaction with at least one chemical site present in the biologically derived material, the biologically derived material including at least one macromolecular biopolymer selected from the group consisting of nucleic acids, proteins, and carbohydrates.

10. The functionalized beta-propiolactone of claims 1, 2, 3,4, 5, 6, 7, 8 or 9 wherein the macromolecular biopolymer is a nucleic acid.

11. The functionalized beta-propiolactone of claims 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 wherein the nucleic acid has at least one at least one substituted purine constituent.

12. The functionalized beta-propiolactone of claims 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 wherein the biologically derived compound is a nucleic acid and wherein the reaction between the nucleic acid and the functionalized beta-propiolactone involves cross linking.

13. A method comprising reacting the functionalized beta-propiolactone of any of claims 1 to 12 with a biologically active material wherein the function of the function of the biologically active material is altered.

14. A method for inducing immune response in a human, wherein the method comprises administering to the human a composition comprising a chemically inactivated virus by treatment with the beta-propiolactone compound claims 1 to 8, the inactivated virus administered in an amount sufficient to induce an immune response in the patient, wherein the wherein the beta-propiolactone compound has a purity greater than 90%.

15. The method of claim 14 wherein the beta-propiolactone compound has a purity greater than 95%.

16. The method of claim 14 wherein the beta-propiolactone compound has a purity greater than 98%.

17. The method of claim 14, 15, 16 wherein the inactivated virus is from the family Picornaviridae or the family Coronaviridae and is one of coronavirus, enterovirus, aphthovirus, cardiovirus, hepatovirus.

18. The method of claims 14, 15, 16 or 17 wherein the inactivated virus is one of the following: coronavirus, aphthovirus, coxsackievirus, rhinovirus.

19. The method of claims 14, 15, 16, 17 or 18 wherein the inactivated virus is SARS-CoV-2.

20. The method of claims 14, 15, 16, 17, 18 or 19 wherein the inactivated virus is present in a composition further comprising a pharmaceutically acceptable carrier and/or excipient.

21. The method of claim 20, wherein the pharmaceutically acceptable carrier and/or excipient is an adjuvant.

22. The method of claims 14, 15, 16, 17, 18, 19, 20 or 21 wherein the inactivated virus is SARS- CoV-2 and the disease against which the immune response is generated is COVID-19.

23. A composition comprising: a beta-propiolactone compound having the general formula:

wherein R¹, R², R³, R⁴ are separately hydrogen or a hydrocarbyl moiety, the hydrocarbyl moiety optionally containing at least one of the following: a heteroatom, at least one substituent; wherein at least one R¹, R², R³, R⁴ are present as a hydrocarbyl moiety; and a carrier medium, the carrier medium including a pharmaceutically acceptable carrier and/or excipient, wherein the beta-propiolactone compound is capable of reacting with at least one biologically active material.

24. The composition of claims 23 wherein the beta-propiolactone is capable of reacting with at least two biologically active materials.

25. The composition of claim 23 or 24 wherein the at least one biologically active material is a nucleic acid.

26. The composition of claims 23, 24 or 25 wherein the beta-propiolactone is capable of reacting with at least two nucleic acids.

27. The compositions of claims 23, 24, 25 or 26 wherein the nucleic acids is selected from the group consisting of adenine, guanine, cytosine, thymine, uracil or mixtures thereof.

28. The composition for reactively cross linking a nucleic acid compound of claim 23, 24, 25,

26 or 27 wherein the pharmaceutically acceptable carrier and/or excipient is an adjuvant.

29. The composition for reactively cross linking at least one nucleic acid compound of claims 23, 24, 25, 26, 27 or 28 wherein the nucleic acid is present in a biological pathogen.

30. The composition for reactively cross linking a nucleic acid compound of claim 29 wherein the biological pathogen is a vims.

31. The composition of claim 23, 23, 24, 25, 26, 27, 28, 29 or 30 wherein the beta functionalized beta-propiolactone compound is the ring open configuration includes at least one of the following general formulae:

wherein each of R¹, R² are hydrogen or a hydrocarbyl moiety, the hydrocarbyl moiety optionally containing at least one heteroatom or at least one substituent, wherein at least one of R¹, R² are present as a hydrocarbyl moiety, and wherein

indicates attachment to a suitable biological structure or intermediate functional group, wherein the at least one biologically active material.

32. An organo-viral complex having at least one of the following general formulae:

wherein each R¹ , R² is hydrogen or a hydrocarbyl moiety, the hydrocarbyl moiety optionally containing at least one heteroatom or at least one substituent, wherein at least one R¹, R² are present as a hydrocarbyl moiety; wherein vims is an infective agent having at least one nucleic acid molecule and a protein coat surrounding at least a portion of the nucleic acid, and wherein n is an integer that is greater than or equal to one.

33. The organo-viral complex of claim 32 wherein n is an integer between 1 and 100.

34. The organo-viral complex of claims 32 or 33 wherein the virus is from the family Picornaviridae or Coronaviridae and is one of coronavirus, enterovirus, aphthovirus, cardiovirus, hepatovirus.

35. The organo-viral complex of claims 32, 33 or 34 wherein the vims is one of the following: coronavirus, coxsackievims, rhinovirus.

36. The organo-viral complex of claims 32, 33, 34 or 35 wherein the vims is SARS-CoV-2.

37. The organo-viral complex of claims 32, 33, 34, 35 or 36 wherein each R¹ , R² are separately and in each occurrence hydrogen or an aryl, substituted aryl, cycloaliphatic, substituted cycloaliphatic, aliphatic or substituted aliphatic moiety.

38. The organo-viral complex of claims 32, 33, 34, 35, 36 or 37 wherein each R¹ , R² are separately and at each occurrence are hydrogen, alkyl, alkenyl, cycloalkyl, heterocyclyl, alkyl heterocyclyl, aryl, aralkyl, alkaryl, heteroaryl, or alkyl heteroaryl, or polyoxyalkylene, or wherein R¹, R² form a 5-20 membered cyclic or heterocyclic ring and wherein R¹ , R² are separately in each occurrence C1-C15 alkyl, C2-C15 alkenyl, C3-C9 cycloalkyl, C2-C20 heterocyclyl, C3-C20 alkyl heterocyclyl, C6-Cis aryl, C7-C25 alkaryl, C7-C25 aralkyl, C5-C18 heteroaryl or C6-C25 alkyl heteroaryl, or polyoxyalkylene.

39. The organo-viral complex of claims 32, 33, 34, 35, 36, 37 or 38 wherein the substituent is halo, alkylthio, alkoxy, hydroxyl, nitro, azido, cyano, acyloxy, carboxy, ester or unsaturated groups two of R¹, R² may be separately in each occurrence C1-C15 alkyl, C3-C6 cycloalkyl, C4- Ci8 heterocyclyl, C4-C18 alkheterocyclyl, C6-C18 aryl, C7-C25 alkaryl, C7-C25 aralkyl, C5-C18 heteroaryl or C6-C25 alkyl heteroaryl.