Substituted organofluoroborates as imaging agents

Fluoridated organofluoroborates comprising at least one 18F atom and precursors thereto, for use in PET scanning.

TECHNICAL FIELD

This invention relates to the field of18F radiolabeled reagents for use in positron emission tomography (PET) imaging.

BACKGROUND

18F is the isotope of choice for many PET cancer imaging applications.

PET imaging agents are often based on a labeled biomolecule. Examples include fluorodeoxyglucose (FDG); Octreotate, an octapeptide that is used to image cancer; and folate, which has been used to image cancer. Since the high energy particle bombardment used to produce18F destroys complex organic molecules,18F is first made as fluoride ion in a cyclotron and subsequently attached to the biomolecule used as the imaging agent. Also, conditions used to incorporate18F are often too harsh for direct labeling of many biomolecules. Therefore,18F is usually introduced into a precursor (such as an aryl fluoride) that is then subsequently appended to a larger molecule. Such multi-step procedures result in delays, with consequent loss in specific radioactivity.

Some methodologies for incorporating18F into imaging agents have been reported, including a new approach which makes use of boron as an acceptor capable of binding several18F atoms, thus increasing the density of positron emitters in the resulting imaging agent (see, for example, PCT publication WO 2005/0077967). In addition, the use of arylboronic acids/esters as18F acceptors has been reported. This approach has circumvented the previous practice of generating aryl fluorides in multi-step procedures.18F radiolabeled substituted aryl-fluoroborates for use in PET imaging have also been reported (see, for example, WO 2009/012596).

SUMMARY

A consideration in the design of PET imaging agents is the longevity of the agent itself. It is desirable that the imaging agent be sufficiently stable with respect to loss of18F ions (termed herein as ‘solvolytic de-18F-fluoridation, as quantified by the half-life at physiological pH of the chemical bond that attaches the 18F-atom to the tracer’). For example, in some applications it is desirable for the imaging agent to have a half-life with respect to solvolytic de-18F-fluoridation at physiological pH that is greater than 10 times the rate of 18F decay. In some applications it is desirable for the imaging agent to have a half-life with respect to solvolytic de-18F-fluoridation at physiological pH of around 1000 minutes or more. It is noted that the rate of solvolytic de-18F-fluoridation is a chemical process that involves bond dissociation, not radioactive decay, and therefore the rate of de-18F-fluoridation is equal to that of non-radioactive defluoridation, where the 18F-fluorine is replaced with a nonradioactive 19F-fluorine atom.

The invention described herein is based, in part, on the discovery that some organofluoroborates exhibit enhanced resistance to solvolytic de-18F-fluoridation and can be useful as PET imaging agents or precursors thereof.

An embodiment makes use of a positron emitting compound or salt thereof, wherein the compound may be of the formula I:

B is boron;

A may be a linear or branched C1-C15alkyl group, a linear or branched C1-C15alkenyl, group, a linear or branched C1-C15alkynyl group, or a C3-C18non-aromatic cycloalkyl group, wherein each of the C1-C15alkyl group, the linear or branched C1-C15alkenyl group, the linear or branched C1-C15alkynyl group, and the C3-C18non-aromatic cycloalkyl group is unsubstituted or substituted and optionally includes at least one heteroatom interposed between two carbon atoms of the carbon chain of the group, wherein each of the at least one heteroatom is independently selected from the group consisting of O, S, N and P;

A may be joined to B through a carbon atom;

each Y1may independently be selected from the group consisting of R1,18F and19F;

Y2may be selected from the group consisting of R2,18F and19F;

R1may be a non-interfering substituent with regard to fluoridation of B;

R2may be a non-interfering substituent with regard to fluoridation of B; and

at least one of (Y1)nand Y2may be18F;

providing that A may be selected such that the pKaof Haof the acid of the formula II:

is less than or equal to about 2.8. When A is substituted at the carbon alpha to the

group with a functional group that has a dissociable proton, the dissociable proton may have a pKa greater than about 9 and contributes to a net positive charge on the functional group. Moieties, A, when selected so that:i) the pKaof Haof the acid of the formula II:

is less than or equal to 2.8; andii) when A is substituted at the carbon alpha to the

group with a functional group that has a dissociable proton, the dissociable proton has a pKa>9 and contributes to a net positive charge on the functional group,
permit the resulting fluoroborate to resist defluoridation. Half-lives with regard to solvolytic de-18F-fluoridation of the resulting organofluoroborates may be at least about 10 times longer or more than the rate of decay of 18F. In other embodiments, half-lives with regard to solvolytic de-18F-fluoridation of the resulting organofluoroborates may be at least about 1000 minutes or more; or at least about 5000 minutes or more; or at least about 10000 minutes or more; or at least about 15000 minutes or more; or at least about 20000 minutes or more; or at least about 25000 minutes or more; or at least about 50000 minutes or more; or at least about 100000 minutes or more; or at least about 125000 minutes or more; or at least about 150000 minutes or more. In some applications where loss of fluorine may be desired (e.g. in comparison of externally bound ligand that loses fluoride to the surrounding bone with internalized ligands that keep the fluoride in a targeted cell) shorter half-lives may be desirable. However, for whole body imaging, longer half-lives are desirable.

In an embodiment, one or more counterions may be present when the compound is charged.

In various embodiments, there is provided a method of making a positron emitting compound or salt thereof, the method comprising contacting an18F source with a compound or salt thereof, wherein the compound may be of the formula I:

B is boron;

A may be a linear or branched C1-C15alkyl group, a linear or branched C1-C15alkenyl, group, a linear or branched C1-C15alkynyl group, or a C3-C18non-aromatic cycloalkyl group, wherein each of the C1-C15alkyl group, the linear or branched C1-C15alkenyl group, the linear or branched C1-C15alkynyl group, and the C3-C18non-aromatic cycloalkyl group is unsubstituted or substituted and optionally includes at least one heteroatom interposed between two carbon atoms of the carbon chain of the group, wherein each of the at least one heteroatom is independently selected from the group consisting of O, S, N and P;

A may be joined to B through a carbon atom;

each Y1may independently be selected from the group consisting of R1and a leaving group that can be displaced by 18F-fluoride;

Y2may be selected from the group consisting of R2and a leaving group that can be displaced by fluoride 18F-fluoride;

at least one of (Y1)nand Y2may be the leaving group when n is 2;

R1may be a non-interfering substituent with regard to fluoridation of B; and

R2may be a non-interfering substituent with regard to fluoridation of B;

providing that A may be selected such that the pKaof Haof the acid of the formula II:

may be less than or equal to 2.8. When A is substituted at the carbon alpha to the

group with a functional group that has a dissociable proton, the dissociable proton may have a pKa greater than about 9 and contributes to a net positive charge on the functional group.

In various embodiments, there is provided a method of performing PET imaging by administering an imaging-effective amount of a positron emitting compound or salt as defined anywhere herein to a subject or object to be subjected to PET.

In various embodiments, there is provided a method of selecting a PET imaging agent or precursor thereof having resistance to solvolytic de-18F-fluoridation at physiological pH, the method comprising:

(i) providing one or more compounds or salt thereof wherein the compound may be of the formula I:

B is boron;

A may be a linear or branched C1-C15alkyl group, a linear or branched C1-C15alkenyl, group, a linear or branched C1-C15alkynyl group, or a C3-C18non-aromatic cycloalkyl group, wherein each of the C1-C15alkyl group, the linear or branched C1-C15alkenyl group, the linear or branched C1-C15alkynyl group, and the C3-C18non-aromatic cycloalkyl group is unsubstituted or substituted and optionally includes at least one heteroatom interposed between two carbon atoms of the carbon chain of the group, wherein each of the at least one heteroatom is independently selected from the group consisting of O, S, N and P;

A may be joined to B through a carbon atom;

each Y1may independently be selected from the group consisting of R1,18F and19F;

Y2may be selected from the group consisting of R2,18F and19F;

R1may be a non-interfering substituent with regard to fluoridation of B;

R2may be a non-interfering substituent with regard to fluoridation of B; and

at least one of (Y1)nand Y2may be18F;

providing that A may be selected such that the pKaof Haof the acid of the formula II:

may be less than or equal to about 2.8;

(ii) assessing the half-life of the presence of the fluorine bound to B; and

(iii) selecting a compound or compounds having said half-life of about 1000 minutes or more as said imaging agent or precursor thereof. When A is substituted at the carbon alpha to the

group with a functional group that has a dissociable proton, the dissociable proton may have a pKa greater than about 9 and contributes to a net positive charge on the functional group

In various embodiments, there is provided a conjugate or salt thereof comprising a peptide conjugated to a positron-emitting compound or salt thereof as described above.

In various embodiments, there is provided a positron emitting compound or salt thereof as described above, or a peptide conjugated to such positron-emitting compound or salt thereof, for use as a PET imaging agent or precursor thereof.

In various embodiments, there is provided a use of a compound or salt thereof as defined above as a precursor in the manufacture of an18F containing PET imaging agent.

In various embodiments, the compound may be of the formula (IV):

DETAILED DESCRIPTION

Any terms not directly defined herein shall be understood to have the meanings commonly associated with them as understood within the art of the invention. Certain terms are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner in describing the devices, methods and embodiments of the invention, and how to make or use them. It will be appreciated that the same thing may be said in more than one way. Consequently, alternative language and synonyms may be used for any one or more of the terms discussed herein. No significance is to be placed upon whether or not a term is elaborated or discussed herein. Some synonyms or substitutable methods, materials and the like are provided. Recital of one or a few synonyms or equivalents does not exclude use of other synonyms or equivalents, unless it may be explicitly stated. Use of examples in the specification, including examples of terms, may be for illustrative purposes only and does not limit the scope and meaning of the embodiments of the invention herein.

“Fluoridation” or “fluorination” are used synonymously herein to refer generally to a chemical reaction by which fluorine is introduced into a compound.

As used herein, the symbol ‘pKa’ is normally understood to a person skilled in the art and refers to the logarithmic constant, pKa, where pKa=−log10Ka. The symbol ‘Ka’ refers to an acid dissociation constant, which is the equilibrium constant for the proton-transfer reaction between a weak acid, HA, and water, H2O. The acid dissociation constant, Ka, for a monoprotic acid, HA, is given by the expression Ka=[A−][H3O+]/[HA].

As used herein, the phrase ‘Cx-Cyalkyl’ group may be used as it is normally understood to a person of skill in the art and often refers to a chemical entity that has a carbon skeleton or main carbon chain comprising a number from x to y (with all individual integers within the range included, including integers x and y) of carbon atoms. For example a ‘C1-C15alkyl’ group may be a chemical entity that has 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 carbon atom(s) in its carbon skeleton or main chain.

As used herein, the term ‘linear’ may be used as it is normally understood to a person of skill in the art and often refers to a chemical entity that comprises a skeleton or main chain that does not split off into more that one contiguous chain. Non-limiting examples of linear alkyls include methyl, ethyl, n-propyl, and n-butyl.

As used herein, the term ‘branched’ may be used as it is normally understood to a person of skill in the art and often refers to a chemical entity that comprises a skeleton or main chain that splits off into more than one contiguous chain. The portions of the skeleton or main chain that split off in more than one direction may be linear, cyclic or any combination thereof. Non-limiting examples of a branched alkyl group include tert-butyl and isopropyl.

A linear or branched C1-C15alkyl group may include a linear or branched saturated C1-C15alkyl group, a linear or branched C2-C15alkenyl group and a linear or branched C2-C15alkynyl group. As used herein, the term ‘saturated’ when referring to a chemical entity may be used as it is normally understood to a person of skill in the art and often refers to a chemical entity that comprises only single bonds. Non-limiting examples of saturated C1-C15alkyl group may include methyl, ethyl, n-propyl, i-propyl, sec-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, i-pentyl, sec-pentyl, t-pentyl, n-hexyl, i-hexyl, 1,2-dimethylpropyl, 2-ethylpropyl, 1-methyl-2-ethylpropyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1,2-triethylpropyl, 1,1-dimethylbutyl, 2,2-dimethylbutyl, 2-ethylbutyl, 1,3-dimethylbutyl, 2-methylpentyl, 3-methylpentyl, sec-hexyl, t-hexyl, n-heptyl, i-heptyl, sec-heptyl, t-heptyl, n-octyl, i-octyl, sec-octyl, t-octyl, n-nonyl, i-nonyl, sec-nonyl, t-nonyl, n-decyl, i-decyl, sec-decyl and t-decyl. Non-limiting examples of C2-C15alkenyl group may include vinyl, allyl, isopropenyl, 1-propene-2-yl, 1-butene-1-yl, 1-butene-2-yl, 1-butene-3-yl, 2-butene-1-yl, 2-butene-2-yl, octenyl and decenyl. Non-limiting examples of C2-C15alkynyl group may include ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl and decynyl. The saturated C1-C15alkyl group, C2-C15alkenyl group or C2-C15alkynyl group may be, for example, and without limitation, interrupted by one or more heteroatoms which may independently be nitrogen, sulfur, oxygen or phosphorus.

As used herein, the term ‘Cx-Cycycloalkyl’ group may be used as it is normally understood to a person of skill in the art and often refers to a compound or a chemical entity in which at least a portion of the carbon skeleton or main chain of the chemical entity may be bonded in such a way so as to form a ‘loop’, circle or ring of atoms that are bonded together. The atoms do not have to all be directly bonded to each other, but rather may be directly bonded to as few as two other atoms in the ‘loop’. As used herein, C3-C18cycloalkyl group may include a non-aromatic C3-C18cycloalkyl group and an aromatic C3-C18cycloalkyl group.

A C3-C18cycloalkyl group may include, for example, and without limitation, a saturated C3-C18cycloalkyl group, a C3-C18cycloalkenyl group, a C3-C18cycloalkynyl group, a C3-C18aryl group, a C3-C18non-aromatic heterocyclic group containing one or more heteroatoms which may independently be nitrogen, sulfur, phosphorus or oxygen, and a C3-C18aromatic heterocyclic group containing one or more heteroatoms which may independently be nitrogen, sulfur, phosphorus or oxygen. Non-limiting examples of the saturated C3-C18cycloalkyl group may include cyclopropanyl, cyclobutanyl, cyclopentanyl, cyclohexanyl, cycloheptanyl, cyclooctanyl, cyclononanyl and cyclodecanyl. Non-limiting examples of the C3-C18cycloalkenyl group may include cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclononanenyl and cyclodecanenyl. Non-limiting examples of the C3-C18aryl group may include phenyl (Ph), pentalenyl, indenyl, naphthyl, and azulenyl.

As used herein, the term ‘substituted’ may be used as it is normally understood to a person of skill in the art and often refers to a chemical entity that has one chemical group replaced with a different chemical group that contains one or more heteroatoms. Unless otherwise specified, a substituted alkyl may be an alkyl in which one or more hydrogen atom(s) may be/are replaced with one or more atom(s) that may be/are not hydrogen(s). For example, chloromethyl may be a non-limiting example of a substituted alkyl, more particularly an example of a substituted methyl. Aminoethyl may be another non-limiting example of a substituted alkyl, more particularly it may be a substituted ethyl.

As used herein, the term ‘unsubstituted’ may be used as it may be normally understood to a person of skill in the art and often refers to a chemical entity that may be a hydrocarbon and/or does not contain a heteroatom. Non-limiting examples of unsubstituted alkyls include methyl, ethyl, tert-butyl, and pentyl.

“Intercepted”, as used herein with respect to the positioning heteroatoms, pertains to a carbon chain in which a heteroatom is interposed between two carbon atoms of the carbon chain.

In an embodiment, the compound may be of the formula I:

In further embodiments, A may be selected such that the pKa of Haof the acid of the formula II:

may be less than or equal to 5.0, less than or equal to 4.5, less than or equal to 4.0, less than or equal to 3.5, less than or equal to 3.0, less than or equal to 2.9, less than or equal to 2.8, less than or equal to 2.7, less than or equal to 2.6, less than or equal to 2.5, less than or equal to 2.4, less than or equal to 2.3, less than or equal to 2.2, less than or equal to 2.1, less than or equal to 2.0, less than or equal to 1.9, less than or equal to 1.8, less than or equal to 1.7, less than or equal to 1.96, less than or equal to 1.5, less than or equal to 1.4, less than or equal to 1.3, less than or equal to 1.2, less than or equal to 1.1, less than or equal to 1.0, less than or equal to 0.9, less than or equal to 0.8, less than or equal to 0.6, less than or equal to 0.4, or less than or equal to 0.2. When A is substituted at the carbon alpha to the

group with a functional group that has a dissociable proton, the dissociable proton has a pKa>9 and contributes to a net positive charge on the functional group.

In further embodiments, A may be selected such that the pKa of Haof the acid of the formula II:

may be from 0.2 to 0.4, 0.2 to 0.6, 0.2 to 0.8, 0.2 to 1.0, 0.2 to 1.2, 0.2 to 1.4, 0.2 to 1.8, 0.2 to 2.0, 0.2 to 2.2, 0.2 to 2.4, 0.2 to 2.6, 0.2 to 2.8, 0.2 to 2.9, 0.4 to 2.4, 0.6 to 2.4, 0.8 to 2.4, 1.0 to 2.4, 1.2 to 2.4, 1.4 to 2.4, 1.6 to 2.4, 1.8 to 2.4, 2.0 to 2.4, or 2.2 to 2.4. When A is substituted at the carbon alpha to the

group with a functional group that has a dissociable proton, the dissociable proton has a pKa>9 and contributes to a net positive charge on the functional group.

In various embodiments, the positron emitting compound or salt may have a solvolytic de-18F-fluoridation half-life at physiological pH of about 1000 minutes or more, about 5000 minutes or more, about 10000 minutes or more, about 15000 minutes or more, about 20000 minutes or more, about 25000 minutes or more, about 50000 minutes or more, about 100000 minutes or more, about 125000 minutes or more, about 150000 minutes or more, or about 200000 minutes or more. In various embodiments, the positron emitting compound or salt may have a solvolytic de-18F-fluoridation half-life at physiological pH from about 1000 minutes to about 200000 minutes, from about 5000 minutes to about 200000 minutes, from about 10000 minutes to about 200000 minutes, from about 15000 minutes to about 200000 minutes, from about 20000 minutes to about 200000 minutes, from about 25000 minutes to about 200000 minutes, from about 50000 minutes to about 200000 minutes, from about 100000 minutes to about 200000 minutes, from about 125000 minutes to about 200000 minutes, or from about 150000 minutes to about 200000 minutes.

In various embodiments, the compound may be of the formula (I), wherein A may be substituted with one or more substituents selected from the group consisting of F, Cl, Br, I, —N+(C1-15alkyl)3, —N+(C2-15alkenyl)3, —N+(C2-15alkynyl)3, —N+H(C1-15alkyl)2, —N+H(C2-15alkenyl)2, —N+H(C2-15alkynyl)2, P+(C1-15alkyl)3, P+(C2-15alkenyl)3, P+(C2-15alkynyl)3, S+(C1-15alkyl)2, S+(C2-15alkenyl)2, S+(C2-15alkynyl)2, oxo (i.e., ═O), OH, —OC1-15alkyl, unsubstituted or substituted aromatic C3-C18cycloalkyl, unsubstituted or substituted non-aromatic C3-C18heterocyclic group, unsubstituted or substituted aromatic C3-C18heterocyclic group, ═NH, —C1-C15alkyl, non-aromatic C3-C18cycloalkyl, —N(C1-15alkyl)2, —N(C2-15alkenyl)2, —N(C2-15alkynyl)2, —COOH, —NH2, —SH, a biomolecule, and a linking group optionally joined to a biomolecule. In various embodiments, A may be substituted with one or more substituents selected from the group consisting of F, Cl, Br, I, —N+(C1-15alkyl)3, —N+(C2-15alkenyl)3, —N+(C2-15alkynyl)3, —N+H(C1-15alkyl)2, —N+H(C2-15alkenyl)2, —N+H(C2-15alkynyl)2, P+(C1-15alkyl)3, P+(C2-15alkenyl)3, P+(C2-15alkynyl)3, oxo (i.e., ═O), OH, unsubstituted or substituted aromatic C3-C18cycloalkyl, unsubstituted or substituted non-aromatic C3-C18heterocyclic group, unsubstituted or substituted aromatic C3-C18heterocyclic group, ═NH, —C1-C15alkyl, non-aromatic C3-C18cycloalkyl, —N(C1-15alkyl)2, —N(C2-15alkenyl)2, —N(C2-15alkynyl)2, —COOH, —NH2, —SH, a biomolecule, and a linking group optionally joined to a biomolecule. In various embodiments, A may be substituted with one or more substituents selected from the group consisting of F, Cl, Br, I, —N+(C1-15alkyl)3, —N+(C2-15alkenyl)3, —N+(C2-15alkynyl)3, —N+H(C1-15alkyl)2, —N+H(C2-15alkenyl)2, —N H(C2-15alkynyl)2, P+(C1-15alkyl)3, P+(C2-15alkenyl)3, P+(C2-15alkynyl)3, oxo (i.e., ═O), OH, unsubstituted or substituted aromatic C3-C18cycloalkyl, unsubstituted or substituted non-aromatic C3-C18heterocyclic group, unsubstituted or substituted aromatic C3-C18heterocyclic group, —C1-C15alkyl, non-aromatic C3-C18cycloalkyl, —N(C1-15alkyl)2, —N(C2-15alkenyl)2, —N(C2-15alkynyl)2, —COOH, —NH2, —SH, a biomolecule, and a linking group optionally joined to a biomolecule. In various embodiments, A may be substituted with at least one substituent that may be a biomolecule or a linking group optionally joined to a biomolecule.

In various embodiments, the compound may be of the formula (IV):

or —R10C═CR11R12; and each of R3, R4, R5, R8, R9, R10, R11, and R12may be as defined anywhere herein. In various embodiments, Q1may be —CR3R4R5, —C≡CR8, or —R10C═CR11R12; and each of R3, R4, R5, R8, R10, R11, and R12may be as defined anywhere herein. In various embodiments, Q1may be —CR3R4R5,

or —R10C═CR11R12; and each of R3, R4, R5, R9, R10, R11, and R12may be as defined anywhere herein. In various embodiments, Q1may be —C≡CR8,

or —R11C═CR11R12; and each of R8, R9, R10, R11, and R12may be as defined anywhere herein. In various embodiments, Q1may be —CR3R4R5or

and each of R3, R4, R5, and R9may be as defined anywhere herein. In various embodiments, Q1may be —CR3R4R5or —R10C═CR11R12; and each of R3, R4, R5, R10, R11and R12may be as defined anywhere herein. In an embodiment, Q1may be —C≡CR8or

and each of R8and R9may be as defined anywhere herein. In an embodiment, Q1may be —C≡CR8or —R10C═CR11R12; and each of R8, R10, R11, and R12may be as defined anywhere herein. In various embodiments, Q1may be

or —R10C═CR11R12; and each of R9, R10, R11, and R12may be as defined anywhere herein. In an embodiment, Q1may be

and R9may be as defined anywhere herein. In an embodiment, Q1may be —R10C═CR11R12; and each of R10, R11, and R12may be as defined anywhere herein. In various embodiments, Q1may be —CR3R4R5, —C≡CR8, or

and each of R3, R4, R5, R8, and R9may be as defined anywhere herein. In an embodiment, Q1may be —CR3R4R5or —C≡CR8; and each of R3, R4, R5, and R8may be as defined anywhere herein. In an embodiment, Q1may be —C≡CR8; and R8may be as defined anywhere herein. In various embodiments, Q1may be —CR3R4R5; and each of R3, R4, and R5may be as defined anywhere herein.

In various embodiments, the compound may be of the formula (IV):

wherein Q1may be —CR3R4R5; and each of R3, R4, and R5may be as defined anywhere herein. Each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —N+R15R16R17, —P+R18R19R20, —S+R21R22, —NR23R24, NHR23, NHR23, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R15, R16, R17, R18, R19, R20, R21, R22, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —N+R15R16R17, —P+R18R19R20, —S+R21R22or —NR23R24; NHR23; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, —S+R21R22, or —NR23R24, and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, —S+R21R22, or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In various embodiments, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —NR15R16R17, —P+R18R19R20, —S+R21R22, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R15, R16, R17, R18, R19, R9, R21and R22may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —N+R15R16R17, —P+R18R19R20, or —S+R21R22; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, or —S+R21R22; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R3, R4and R5may independently be H, D, F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, or —S+R21R22; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In various embodiments, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —N+R15R16R17, —P+R18R19R20, —NR23R24, NHR23, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R15, R16, R17, R18, R19, R20, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —N+R15R16R17, —P+R18R19R20, or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, or NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —N+R15R16R17, —S+R21R22, —NR23R24, NHR23, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R15, R16, R17, R21, R22, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —N+R15R16R17, —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —N+R15R16R17, —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —N+R15R16R17, —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In various embodiments, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —P+R18R19R20, —S+R21R22, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R18, R19, R20, R21, R22, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —P+R18R19R20, —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R18, R19, R20, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —P+R18R19R20, —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R18, R19, R20, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —P+R18R19R20, —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R18, R19, R20, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, —N+R15R16R17, —P+R18R19R20, —S+R21R22, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R15, R16, R17, R18, R19, R20, R21, R22, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, N+R15R16R17, —P+R18R19R20, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R15, R16, R17, R18, R19, and R20may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —N+R15R16R17, or —P+R18R19R20; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, and R20may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —N+R15R16R17, or —P+R18R19R20; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, and R20may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —N+R15R16R17, or —P+R18R19R20; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, and R20may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In various embodiments, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —NR15R16R17, —S+R21R22, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R15, R16, R17, R21and R22may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —N+R15R16R17, or —S+R21R22; and at least one of R3, R4, R5, R15, R16, R17, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —N+R15R16R17, or —S+R21R22; and at least one of R3, R4, R5, R15, R16, R17, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —N+R15R16R17, or —S+R21R22; and at least one of R3, R4, R5, R15, R16, R17, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —N+R15R16R17, —NR23R24, NHR23, a biomolecule, a linking group optionally joined to, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R15, R16, R17, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —N+R15R16R17, or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —N+R15R16R17, —NR23R24, or NHR23; and at least one of R3, R4, R5, R15, R16, R17, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —N+R15R16R17, or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In various embodiments, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —P+R18R19R20, —S+R21R22, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R18, R19, R20, R21and R22may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —P+R18R19R20, or —S+R21R22; and at least one of R3, R4, R5, R18, R19, R20, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —P+R18R19R20, or —S+R21R22; and at least one of R3, R4, R5, R18, R19, R20, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —P+R18R19R20, or —S+R21R22; and at least one of R3, R4, R5, R18, R19, R20, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In various embodiments, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —P+R18R19R20, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R18, R19, R20, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —P+R18R19R20, or —NR23R24and at least one of R3, R4, R5, R18, R19, R20, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —P+R18R19R20, or —NR23R24and at least one of R3, R4, R5, R18, R19, R20, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —P+R18R19R20, or —NR23R24; and at least one of R3, R4, R5, R18, R19, R20, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In various embodiments, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —S+R21R22, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R21, R22, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, —N+R15R16R17, —P+R18R19R20, —S+R21R22, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R15, R16, R17, R18, R19, R20, R21and R22may be as defined anywhere herein; at least one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, or —S+R21R22; and at least one of R3, R4, R15, R16, R17, R18, R19, R20, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, or —S+R21R22; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, —N+R15R16R17, —P+R18R19R20, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R15, R16, R17, R18, R19, R20, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be —N+R15R16R17, —P+R18R19R20, or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R3, R4and R5may be —N+R15R16R17, —P+R19R20, or —NR23R24and at least one of R3, R4, R5, R15, R16, R17, R18, R19, R20, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, —N+R15R16R17, —S+R21R22, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R15, R16, R17, R21, R22, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be —N+R15R16R17, —S+R21R22or —NR23R24; and at least one of R3, R4, R15, R16, R17, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R3, R4and R5may be —N+R15R16R17, —S+R21R22or —NR23R24and at least one of R3, R4, R5, R15, R16, R17, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In various embodiments, each of R3, R4, and R5may independently be H, D, —P+R18R19R20, —S+R21R22, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R18, R19, R20, R21, R22, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be —P+R18R19R20, —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R18, R19, R20, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, one of R3, R4and R5may be —P+R18R19R20, —S+R21R22or —NR23R24and at least one of R3, R4, R5, R18, R19, R20, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —P+R18R19R20, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R18, R19, and R20may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, or —P+R18R19R20; and at least one of R3, R4, R5, R18, R19, and R20may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —P+R18R19R20; and at least one of R3, R4, R5, R18, R19, and R20may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —P+R18R19R20; and at least one of R3, R4, R5, R18, R19, and R20may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In various embodiments, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —S+R21R22, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R21and R22may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, or —S+R21R22; and at least one of R3, R4, R5, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —S+R21R22; and at least one of R3, R4, R5, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —S+R21R22; and at least one of R3, R4, R5, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, or —NR23R24; and at least one of R3, R4, R5, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —NR23R24; and at least one of R3, R4, R5, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —NR23R24; and at least one of R3, R4, R5, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, —N+R15R16R17, —P+R18R19R20, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R15, R16, R17, R18, R19, and R20may be as defined anywhere herein; at least one of R3, R4and R5may be —N+R15R16R17or —P+R18R19R20; and at least one of R3, R4, R5, R15, R16, R17, R18, R19, and R20may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R3, R4and R5may be —N+R15R16R17or —P+R18R19R20and at least one of R3, R4, R5, R15, R16, R17, R18, R19, and R20may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, —N+R15R16R17, —S+R21R22, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R15, R16, R17, R21and R22may be as defined anywhere herein; at least one of R3, R4and R5may be —N+R15R16R17or —S+R21R22; and at least one of R3, R4, R5, R15, R16, R17, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R3, R4and R5may be —N+R15R16R17or —S+R21R22and at least one of R3, R4, R5, R15, R16, R17, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In various embodiments, each of R3, R4, and R5may independently be H, D, —N+R15R16R17, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R15, R16, R17, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be —N+R15R16R17or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R3, R4and R5may be —N+R15R16R17or —NR23R24; and at least one of R3, R4, R5, R15, R16, R17, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, —P+R18R19R20, —S+R21R22, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R18, R19, R20, R21and R22may be as defined anywhere herein; at least one of R3, R4and R5may be —P+R18R19R20or —S+R21R22; and at least one of R3, R4, R5, R18, R19, R20, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, one of R3, R4and R5may be —P+R18R19R20or —S+R21R22; and at least one of R3, R4, R5, R18, R19, R20, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, —P+R18R19R20, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R18, R19, R20, R23, and R24may be as defined anywhere herein; at least one of R3, R4and R5may be —P+R18R19R20or —NR23R24; and at least one of R3, R4, R5, R18, R19, R20, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, one of R3, R4and R5may be —P+R18R19R20or —NR23R24; and at least one of R3, R4, R5, R18, R19, R20, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, —S+R21R22, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R21, R22, R23and R24may be as defined anywhere herein; at least one of R3, R4and R5may be —S+R21R22or —NR23R24; and at least one of R3, R4, R5, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, one of R3, R4and R5may be —S+R21R22or —NR23R24and at least one of R3, R4, R5, R21, R22, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule.

In an embodiment, each of R3, R4, and R5may independently be H, D, F, Cl, Br, I, CX3, —N+R15R16R17, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each X may be the same or different and may be F, Cl, Br, or I; each of R15, R16and R17may be as defined anywhere herein; at least one of R3, R4and R5may be F, Cl, Br, I, CX3, or —N+R15R16R17; and at least one of R3, R4, R5, R15, R16and R17may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or at least one of R3, R4and R5may be —N+R15R16R17; and at least one of R3, R4, R5, R15, R16and R17may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R3, R4and R5may independently be F, Cl, Br, I, or CX3, or one of R3, R4and R5may be —N+R15R16R17; and at least one of R3, R4, R5, R15, R16and R17may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In various embodiments, the compound may be of the formula (IV):

In various embodiments, the compound may be of the formula (IV):

wherein Q1may be —CR3R4R5; and each of R3, R4, and R5may independently be H, D, —N+R15R16R17, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R15, R16and R17may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P, each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the nitrogen atom of —N+R15R16R17through a carbon atom, or R15may be absent and R16and R17may be joined so that —N+R15R16R17forms a positively charged nitrogen containing heterocyclic group which may be substituted or unsubstituted; at least one of R3, R4and R5may be —N+R15R16R17; and at least one of R3, R4, R5, R15, R16, and R17may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, R3may be —N+R15R16R17; each of each of R4and R5may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R15, R16and R17may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P, each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the nitrogen atom of —N+R15R16R17through a carbon atom, or R15may be absent and R16and R17may be joined so that —N+R15R16R17forms a positively charged nitrogen containing heterocyclic group which may be substituted or unsubstituted; and at least one of R4, R5, R15, R16and R17may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In an embodiment, each of R15, R16and R17may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P, and each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the nitrogen atom of —N+R15R16R17through a carbon atom. In various embodiments, R15may be absent and R16and R17may be joined so that —N+R15R16R17forms a positively charged nitrogen containing heterocyclic group which may be substituted or unsubstituted. In various embodiments, R15may be absent and R16and R17may be joined so that —NR15R16R17forms a positively charged 4 to 6-membered nitrogen containing heterocyclic group which may be substituted or unsubstituted. In an embodiment, the positively charged 4 to 6-membered nitrogen containing heterocyclic group may be a pyridinium group, an imidazolium group, a pyrazinium group, a pyrimidinium group or a pyridazinium group, each of which may be substituted or unsubstituted. In various embodiments, the positively charged nitrogen containing heterocyclic group may be unsubstituted. In various embodiments, the positively charged nitrogen containing heterocyclic group which may be substituted with one or more substituents that are selected from the group consisting of a C1-C15alkyl group, a biomolecule, a linking group optionally joined to a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, the compound may be of the formula (IV):

wherein Q1may be —CR3R4R5; and each of R3, R4, and R5may independently be H, D, —P+R18R19R20, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R18, R19, and R20may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the phosphorus atom of —P+R18R19R20through a carbon atom; at least one of R3, R4and R5may be —P+R18R19R20; and at least one of R3, R4, R5, R18, R19, and R20may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, R3may be —P+R18R19R20; each of R4and R5may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R18, R19and R20may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the phosphorus atom of —P+R18R19R20through a carbon atom; and at least one of R4, R5, R18, R19, and R20may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, the compound may be of the formula (IV):

wherein Q1may be —CR3R4R5; and each of R3, R4, and R5may independently be H, D, —S+R21R22, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R21and R22may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the sulfur atom of —S+R21R22through a carbon atom; at least one of R3, R4and R5may be —S+R21R22; and at least one of R3, R4, R5, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, R3may be —S+R21R22; each of R4and R5may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R21and R22may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the sulfur atom of —S+R21R22through a carbon atom; and at least one of R4, R5, R21, and R22may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, the compound may be of the formula (IV):

wherein Q1may be —CR3R4R5; and each of R3, R4, and R5may independently be H, D, —NR23R24, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R23and R24may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the nitrogen atom of —NR23R24through a carbon atom; at least one of R3, R4and R5may be —NR23R24; and at least one of R3, R4, R5, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, R3may be —NR23R24; each of R4and R5may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —CR3R4R5through a carbon atom; each of R23and R24may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the nitrogen atom of —NR23R24through a carbon atom; and at least one of R4, R5, R23, and R24may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, the compound may be of the formula (IV):

wherein Q1may be —C≡CR8; and R8may be as defined anywhere herein.

a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be substituted with at least one substituent that may be a biomolecule and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —C≡CR8through a carbon atom; each of R25, R26, R27, R28, R29, R30, R31, R32, R33, R34, and R35may be as defined anywhere herein; and at least one of R8, R25, R26, R27, R28, R29, R30, R31, R32, R33, R34, and R35may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule.

In various embodiments, R8may be —N+R25R26R27; each of R25, R26and R27may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the nitrogen atom of —N+R25R26R27through a carbon atom, or R25may be absent and R26and R27may be joined so that —N+R25R26R27forms a positively charged nitrogen containing heterocyclic group which may be unsubstituted or substituted; and at least one of R25, R26and R27may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In an embodiment, the positively charged 4 to 6-membered nitrogen containing heterocyclic group may be a pyridinium group, an imidazolium group, a pyrazinium group, a pyrimidinium group or a pyridazinium group, each of which may be substituted or unsubstituted. In various embodiments, the positively charged nitrogen containing heterocyclic group may be unsubstituted. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, R8may be —P+R28R29R30; each of R28, R29, and R30may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the phosphorus atom of —P+R28R29R30through a carbon atom; and at least one of R28, R29and R30may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, R8may be —S+R31R32; each of R31and R32may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the sulfur atom of —S+R31R32through a carbon atom; and at least one of R31and R32may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In an embodiment, R8may be —NR33R34; each of R33and R34may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the nitrogen atom of —NR33R34through a carbon atom; and at least one of R33and R34may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In an embodiment, R8may be

and R35may be a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be substituted with at least one substituent that may be a biomolecule and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; and each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of

through a carbon atom. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, R8may be a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be substituted with at least one substituent that may be a biomolecule and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; and each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —C≡CR8through a carbon atom. In various embodiments, R8may be a linear or branched C1-C15alkyl group or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be substituted with at least one substituent that may be a biomolecule and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; and each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of —C≡CR8through a carbon atom. In various embodiments, R8may be a biomolecule or a linking group optionally joined to a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In an embodiment, the compound may be of the formula (IV):

wherein Q1may be

and R9may be as defined anywhere herein.

In various embodiments, R9may be —NR38R39,

a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be substituted with at least one substituent that may be a biomolecule and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of

through a carbon atom; each of R38, R39, and R40may be as defined anywhere herein; and at least one of R9, R38, R39, and R40may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, R9may be —NR38R39; each of R38and R39may independently be H, D, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be unsubstituted or substituted and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the nitrogen atom of —NR38R39through a carbon atom; and at least one of R38and R39may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that may be substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In an embodiment, R9may be

and R40may be a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be substituted with at least one substituent that may be a biomolecule and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; and each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of

through a carbon atom. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, R9may be a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be substituted with at least one substituent that may be a biomolecule and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; and each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of

through a carbon atom. In an embodiment, R9may be a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be substituted with at least one substituent that may be a biomolecule and may be optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; and each of the C1-C15alkyl group and the C3-C18cycloalkyl group may be joined to the carbon atom of

through a carbon atom. In an embodiment, R9may be a biomolecule or a linking group optionally joined to a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, the compound may be of the formula (IV):

wherein Q1may be —R10C═CR11R12; and each of R10, R11, and R12may be as defined anywhere herein.

In various embodiments, each of R10, R11, and R12may independently be H, D, F, Cl, Br, I, CX3, —N+R41R42R43, —P+R44R45R46, —S+R47R48, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom, and each X is the same or different and is F, Cl, Br, or I; each of R41, R42, R43, R44, R45, R46, R47, and R48may be as defined anywhere herein; at least one of R10, R11, and R12may independently be F, Cl, Br, I, CX3, —N+R41R42R43, —P+R44R45R46, or —S+R47R48; and at least one of R10, R11, R12, R41, R42, R43, R44, R45, R46, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or at least one of R10, R11, and R12may be —N+R41R42R43, —P+R44R45R46, or —S+R47R48; and at least one of R10, R11, R12, R41, R42, R43, R44, R45, R46, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or one of R10, R11, and R12may be —N+R41R42R43, —P+R44R45R46, or —S+R47R48; and at least one of R10, R11, R12, R41, R42, R43, R44, R45, R46, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, F, Cl, Br, I, CX3, —N+R41R42R43, —P+R44R45R46, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom, and each X is the same or different and is F, Cl, Br, or I; each of R41, R42, R43, R44, R45, R46, R47, and R48may be as defined anywhere herein; at least one of R10, R11, and R12may independently be F, Cl, Br, I, CX3, —N+R41R42R43or —P+R44R45R46; and at least one of R10, R11, R12, R41, R42, R43, R44, R45, and R46may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or at least one of R10, R11, and R12may be —N+R41R42R43, or —P+R44R45R46; and at least one of R10, R11, R12, R41, R42R43, R44, R45, and R46may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or one of R10, R11, and R12may be —N+R41R42R43, or —P+R44R45R46; and at least one of R10, R11, R12, R41, R42, R43, R44, R45, and R46may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, F, Cl, Br, I, CX3, —N+R41R42R43, —S+R47R48, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom, and each X is the same or different and is F, Cl, Br, or I; each of R41, R42, R43, R47, and R48may be as defined anywhere herein; at least one of R10, R11, and R12may independently be F, Cl, Br, I, CX3, —N+R41R42R43, or —S+R47R48and at least one of R10, R11, R12, R41, R42, R43, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or at least one of R10, R11, and R12may be —N+R41R42R43or —S+R47R48; and at least one of R10, R11, R12, R41, R42, R43, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or one of R10, R11, and R12may be —N+R41R42R43or —S+R47R48; and at least one of R10, R11, R12, R41, R42, R43, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, F, Cl, Br, I, CX3, —P+R44R45R46, —S+R47R48, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom, and each X is the same or different and is F, Cl, Br, or I; each of R44, R45, R46, R47, and R48may be as defined anywhere herein; at least one of R10, R11, and R12may independently be F, Cl, Br, I, CX3, —P+R44R45R46, or —S+R47R48; and at least one of R10, R11, R12, R44, R45, R46, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or at least one of R10, R11, and R12may be —P+R44R45R46or —S+R47R48; and at least one of R10, R11, R12, R44, R45, R46, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or one of R10, R11, and R12may be —P+R44R45R46or —S+R47R48; and at least one of R10, R11, R12, R44, R45, R46, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, —N+R41R42R43, —P+R44R45R46, —S+R47R48, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom; each of R41, R42, R43, R44, R45, R46, R47, and R48may be as defined anywhere herein; at least one of R10, R11, and R12may independently be —N+R41R42R43, —P+R44R45R46, or —S+R47R48; and at least one of R10, R11, R12, R41, R42, R43, R44, R45, R46, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R10, R11, and R12may be —N+R41R42R43, —P+R44R45R46, or —S+R47R48; and at least one of R10, R11, R12, R41, R42, R43, R44, R45, R46, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, F, Cl, Br, I, CX3, —N+R41R42R43, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom, and each X is the same or different and is F, Cl, Br, or I; each of R41, R42, and R43may be as defined anywhere herein; at least one of R10, R11, and R12may independently be F, Cl, Br, I, CX3, or —N+R41R42R43; and at least one of R10, R11, R12, R41, R42, and R43may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, at least two of R10, R1, and R12may independently be F, Cl, Br, I, or CX3, or at least one of R10, R11, and R12may be —N+R41R42R43; and at least one of R10, R11, R12, R41, R42, and R43may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or one of R10, R11, and R12may be —N+R41R42R43; and at least one of R10, R11, R12, R41, R42, and R43may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, F, Cl, Br, I, CX3, —P+R44R45R46, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom, and each X is the same or different and is F, Cl, Br, or I; each of R4, R45, and R46may be as defined anywhere herein; at least one of R10, R11, and R12may independently be F, Cl, Br, I, CX3, or —P+R44R45R46; and at least one of R10, R11, R12, R44, R45, and R46may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, at least two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or at least one of R10, R11, and R12may be —P+R44R45R46; and at least one of R10, R11, R12, R44, R45, and R46may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or one of R10, R11, and R12may be —P+R44R45R46; and at least one of R10, R11, R12, R44, R45, and R46may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, F, Cl, Br, I, CX3, —S+R47R48, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom, and each X is the same or different and is F, Cl, Br, or I; each of R47and R48may be as defined anywhere herein; at least one of R10, R11, and R12may independently be F, Cl, Br, I, CX3, or —S+R47R48; and at least one of R10, R11, R12, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, at least two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or at least one of R10, R11, and R12may be —S+R47R48; and at least one of R10, R11, R12, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, at two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3, or one of R10, R11, and R12may be —S+R47R48; and at least one of R10, R11, R12, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, —N+R41R42R43, —P+R44R45R46, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom; each of R41, R42, R43, R44, R45, and R46may be as defined anywhere herein; at least one of R10, R11, and R12may independently be —N+R41R42R43or —P+R44R45R46; and at least one of R10, R11, R12, R41, R42, R43, R44, R45, and R46may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R10, R11, and R12may be —NR41R42R43or —P+R44R45R46; and at least one of R10, R11, R12, R41, R42, R43, R44, R45, and R46may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, —N+R41R42R43, —S+R47R48, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom; each of R41, R42, R43, R47, and R48may be as defined anywhere herein; at least one of R10, R11, and R12may independently be —N+R41R42R43or —S+R47R48; and at least one of R10, R11, R12, R41, R42, R43, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R10, R11, and R12may be —N+R41R42R43or —S+R47R48; and at least one of R10, R11, R2, R41, R42, R43, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, —P+R44R45R46, —S+R47R48, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom; each of R44, R45, R46, R47, and R48may be as defined anywhere herein; at least one of R10, R11, and R12may independently be —P+R44R45R46or —S+R47R48; and at least one of R10, R11, R12, R44, R45, R46, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, one of R10, R11, and R12may be —P+R44R45R46or —S+R47R48; and at least one of R10, R11, R12, R4, R45, R46, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, F, Cl, Br, I, CX3, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR10R12through a carbon atom, and each X is the same or different and is F, Cl, Br, or I; at least one of R10, R11, and R12may independently be F, Cl, Br, I, or CX3; and at least one of R10, R11, and R12may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, two of R10, R11, and R12may independently be F, Cl, Br, I, or CX3; and one of R10, R11, and R12may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, —N+R41R42R43, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom; each of R41, R42, and R43may be as defined anywhere herein; at least one of R10, R11, and R12may independently be —N+R41R42R43; and at least one of R10, R11, R12, R41, R42, and R43may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In various embodiments, one of R10, R11, and R12may be —N+R41R42R43; and at least one of R10, R11, R12, R41, R42, and R43may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule, or forms a group that is substituted with at least one substituent that is a biomolecule. In an embodiment, the positively charged 4 to 6-membered nitrogen containing heterocyclic group may be a pyridinium group, an imidazolium group, a pyrazinium group, a pyrimidinium group or a pyridazinium group, each of which may be substituted or unsubstituted. In various embodiments, the positively charged nitrogen containing heterocyclic group may be unsubstituted. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, —P+R44R45R46, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom; each of R44, R45, and R46may be as defined anywhere herein; at least one of R10, R11, and R12may independently be —P+R44R45R46; and at least one of R10, R11, R12, R44, R45, and R46may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, one of R10, R11, and R12may be —P+R44R45R46; and at least one of R10, R11, R12, R44, R45, and R46may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R10, R11, and R12may independently be H, D, —S+R47R48, a biomolecule, a linking group optionally joined to a biomolecule, a linear or branched C1-C15alkyl group, or a C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P; each of the C1-C15alkyl group and the C3-C18cycloalkyl group is joined to the carbon atom of —R10C═CR11R12through a carbon atom; each of R47and R48may be as defined anywhere herein; at least one of R10, R11, and R12may independently be —S+R47R48; and at least one of R10, R11, R12, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, one of R10, R11, and R12may be —S+R47R48; and at least one of R10, R11, R12, R47, and R48may be a biomolecule, a linking group optionally joined to a biomolecule, or a group that is substituted with at least one substituent that may be a biomolecule. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

The term ‘biomolecule’ as used herein refers to a biomolecule, or analog or derivative of a biomolecule, or other molecule that may be delivered into a human or animal in order to track or image distribution of the biomolecule within a human or animal body or tissue via positron emission tomography. Examples are disclosed in WO 2005/077967. In some embodiments, a ‘biomolecule’ refers to any molecule of medical, physiological or scientific significance, analog or derivative thereof that may be compatible with a biological system or which possesses biological activity. Biomolecules may be delivered into a human or animal and include biomolecules that become localized at particular places in the organism. Examples include sugars, amino acids, nucleic acids, nucleotides, nucleosides, peptide hormones (steroid and nonsteroid), antibodies, aptamers and oligonucleotides, proteins, peptides, oligonucleotides, lipids, hormones, drugs (synthetic drugs and natural products), polysaccharides, liposomes, micelles, microsomes, magnetic particles, metal chelators, oligoribonucleotides, oligonucleotides and related analogs bearing modifications in the backbone, nucleobase, or phosphate linker regions that enhance stability or modulate specificity, peptidomimetics, dendrimers, drug delivery agents, nanotubes, fullerenes, virus particles, and other targeting molecules (e.g. cancer targeting molecules). Specific examples include, but are not limited to, biotin, matrix mettaloprotease inhibitors such as marimastat, insulin, somatostatin, somatotropin, somatomedin, adrenocorticotropic hormone, parathormone, follicle stimulating hormone, luteinizing hormone, epidermal growth factor, thyroid stimulating hormone, thyroid stimulating hormone releasing hormone, luteinizing hormone releasing hormone, vasopressin, bombesin, endothelin, gonadotropins, gonadotropin releasing hormone, antiflamin I&II, NLE-antiflamin II, brain natriureitic peptide, calcitonin, corticotropin releasing peptide, oxytocin, calpain inhibitor peptide, alpha-CGRP, corticotropin releasing factor, galanin, growth hormone releasing factor, guanylin, alpha-helical corticotropin releasing factor, laminin, alpha-melanocyte stimulating hormone, platelet derived growth factor, neuromedin, neurotensin, pancreatic polypeptide, pentagastrin, peptide-YY, pituitary adenylate cyclase activating peptide, secretin, thyrotropin releasing hormone, urocortin, vasoactive intestinal peptide, vasopressin, vascular endothelial growth factor, apamin, bungarotoxin, calciceptin, charybdotoxin, cobrotoxin, conotoxin, dendrotoxin, melittin, neuropeptide-Y, imperatoxin, taycatoxin, annexin, inhibin, insulin-like growth factor, prolactin, melanin stimulating hormone, melanin concentrating hormone, substance-P, tachykinin, angiotensin, antibodies of general structural classes of IgG, IgM, IgE, IgA, as well as single-chain, monoclonal, and recombinant forms used for current and anticipated imaging, diagnostic, and therapeutic applications. Specific targets that can be recognized by antibodies comprise without limitation: melanoma cell, melanoma specific antigen, myelin basic protein, breast cancer specific tumor markers such as Her2-Neu and Brc-Ab1, alpha-fetoprotein, human chorionic gonadotropin, prostate specific antigen, prostate specific membrane antigen, epidermal growth factor receptors, fibroblast growth factor receptor, insulin receptor. Other examples are antibodies approved for use in therapy such as Herceptin™ (Amgen), Erbitux™ (Imclone). Polymers containing nucleobases and nucleotides including RNA, DNA, and PNAs and various synthetic derivatives thereof that reflect modification of the sugar, intemucleoside linkage (backbone) and nucleobase portions are also contemplated. Oligonucleotides that can be used for imaging, for example: antisense oligonucleotides that target mRNA of genes implicated in the disease state, siRNA or RNAi molecules that target mRNA via RNA silencing, and aptamer structures which represent a diverse class of folded nucleic acid structures that target protein or glycoforms of proteins or both, or folded RNA structures. Further examples are aptamers approved for clinical use or those intended for clinical and diagnostic use such as Macugen™ (Eyetech) and aptamers that are used in the context of surface arrayed aptamers for diagnostic purposes, oligosaccharides of both synthetic and natural origin that are found on the surface of cellular receptors or can mimic the glycoforms of cellular receptors and proteins. Other saccharide components in synthetic glycoforms are sialic acid, mannose, fucose, N-acetyl-glucosamine, N-acetyl-mannosamine, maltose, galactose and N-acetyl-galactosamine, small to mid-size molecular weight ligands for proteins comprise various classes of compounds, for example: porphyrins, lectins, lipids, steroids, barbiturates, taxanes, terpenes, terpenoids, canabinoids, opioids, nucleosides, purines, pyrimidines, heteroaromatics, quinolines, biogenic amines, amino acids, indole-alkaloids, topane alkaloids, statins, enzyme inhibitors, nonsteroidal anti-inflammatory agents, monosaccharides, folates, derivatives of folate, methotrexate, derivatives of methotrexate, trexates, vitamins, growth hormone, VEGF, EGF, an antibody, a breast cancer antigen specific antibody, a prostate cancer antigen specific antibody, a melanoma antigen specific antibody, a ligand, a RGD-motif ligand recognizing a matrix metalloprotease, an aptamer, an aptamer recognizing a cell surface protein, folic acid, a folic acid derivative and a methotrexate. Tracer molecules used in this invention may be conjugated to a ligand such as a biomolecule that preferentially interacts with a tissue type or cell type of interest. In some embodiments, a precursor substituted alkyl-boronic acid may be pre-conjugated to a biomolecule of interest and subsequently fluoridated when needed in a one-step aqueous fluoridation reaction. A typical reaction may occur in a buffered solution of KHF2where the18F may be generated in carrier free form and supplemented with carrier19F either at the time of fluoridation or during a chase reaction, while in another typical reaction the18F will be used in an isotope exchange reaction. In various embodiments, the biomolecule may be a sugar, a peptide, a nucleic acid, a lipid, a steroid, a biogenic amine or derivative or analog thereof. In various embodiments, the biomolecule may be a hormone, a drug, insulin, somatostatin, growth hormone, VEGF, EGF, bombesin, a gonadotropin, gonadotropin releasing hormone, corticotropin releasing peptide, oxytocin, corticotropin releasing factor, growth hormone releasing factor, platelet derived growth factor, neurotensin, urocortin, vasoactive intestinal peptide, inhibin, insulin-like growth factor, an antisense oligonucleotide that targets mRNA of a gene implicated in the disease state, a siRNA or RNAi molecule that targets mRNA via RNA silencing, an antibody, a breast cancer antigen specific antibody, a prostate cancer antigen specific antibody, a melanoma antigen specific antibody, a ligand, a RGD-motif ligand recognizing a matrix metalloprotease, an aptamer, an aptamer recognizing a cell surface protein, folic acid, a folic acid derivative and a methotrexate or a derivative or analog thereof.

Linking groups may include aliphatic or aromatic moieties designed to insulate the biomolecule from the boron atom by an appropriate distance or to ensure that appropriate atoms are adjacent the boron atom to facilitate the fluoridation process. Groups which facilitate subsequent addition of a biomolecule are well known in the art and may include moieties which readily form a bond to a selected biomolecule, a variety of such groups being known in the art. These include thiol and amine reactive groups and other such groups which may be useful for joining a compound of this invention to functionalities on biomolecules including hydroxide, carboxylic acid, amine, sulfhydryl groups, etc. Further contemplated herein may be the joining of a compound of this invention to a biomolecule through other linkages involving what is known as “click chemistry” with examples that include reactions of tetrazines with either strained alkenes or alkynes, or alkynes with azides either by metal-mediated catalysis or strain promotion. Alternatively, bonds other than covalent bonds are contemplated. Thus, groups, which provide for ionic, hydrophobic and other non-covalent linkages to a biomolecule are contemplated. Exemplary linker groups for facilitating conjugation to a biomolecule include:

In various embodiments, n may be 1 or 2; and each Y1may independently be selected from the group consisting of R1,18F, and19F. In various embodiments, n may be 1 or 2. Each Y1may independently be selected from the group consisting of R1,18F, and19F. In various embodiments, n may be 2 and each Y1may independently be selected from the group consisting of R1,18F, and19F. In various embodiments, n may be 2, and each Y1may independently be R1or18F, each Y1may independently be R1or19F, or each Y1may independently be18F or19F. In various embodiments, n may be 2, and each Y1may be R1, each Y1may be18F, or each Y1may be19F. In various embodiments, n may be 1, and Y1may be R1,18F, or19F.

In various embodiments, R1may be absent or present. In an embodiment, when R1is present, R1may be any non-interfering group with regard to fluoridation of B. In an embodiment, R1may be a linear or branched C1-C15alkyl group or a non-aromatic C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the non-aromatic C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P. In an embodiment, the C1-C15alkyl group or non-aromatic C3-C18cycloalkyl group may be intercepted with at least one heteroatom at or near the attachment point to B. In particular embodiments, R1may be a linear or branched C1-C6alkyl group or a non-aromatic C3-C8cycloalkyl group. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the non-aromatic C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, Y2may independently be selected from the group consisting of R2,18F, and19F. In an embodiment, Y2may independently be selected from the group consisting of R2and18F. In an embodiment, Y2may independently be selected from the group consisting of R2and19F. In an embodiment, Y2may independently be selected from the group consisting of18F and19F. In various embodiments, Y2may be R2. In various embodiments, Y2may be18F. In various embodiments, Y2may be19F.

In various embodiments, R2may be absent or present. In an embodiment, when R2is present, R2may be any non-interfering group with regard to fluoridation of B. In an embodiment, R2may be a linear or branched C1-C15alkyl group or a non-aromatic C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the non-aromatic C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P. In an embodiment, the C1-C15alkyl group or non-aromatic C3-C18cycloalkyl group may be intercepted with at least one heteroatom at or near the attachment point to B. In particular embodiments, R2may be a linear or branched C1-C6alkyl group or a non-aromatic C3-C8cycloalkyl group. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the non-aromatic C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, each of R1and R2may independently be a linear or branched C1-C15alkyl group or a non-aromatic C3-C18cycloalkyl group, wherein each of the C1-C15alkyl group and the non-aromatic C3-C18cycloalkyl group is unsubstituted or substituted and is optionally intercepted with at least one heteroatom selected from the group consisting of O, S, N and P. In various embodiments, the C1-C15alkyl group may be a C1-C6alkyl group. In various embodiments, the non-aromatic C3-C18cycloalkyl group may be a C4-C8cycloalkyl group.

In various embodiments, n may be 2; each Y1may independently be18F or19F; Y2may be18F or19F; and at least one of (Y1)nand Y2may be18F.

In various embodiments, the a linear or branched C1-C15alkyl group may be a linear or branched saturated C1-C2alkyl group, or a linear or branched saturated C1-C3alkyl group, or a linear or branched saturated C1-C4alkyl group, or a linear or branched saturated C1-C5alkyl group, or a linear or branched saturated C1-C6alkyl group, or a linear or branched saturated C1-C7alkyl group, or a linear or branched saturated C1-C8alkyl group, or a linear or branched saturated C1-C9alkyl group, or a linear or branched saturated C1-C10alkyl group, or a linear or branched saturated C1-C1alkyl group, or a linear or branched saturated C1-C12alkyl group, or a linear or branched saturated C1-C13alkyl group, or a linear or branched saturated C1-C14alkyl group, or a linear or branched saturated C1-C15alkyl group, or a linear or branched saturated C1alkyl group, or a linear or branched saturated C2alkyl group, or a linear or branched saturated C3alkyl group, or a linear or branched saturated C4alkyl group, or a linear or branched saturated C5alkyl group, or a linear or branched saturated C6alkyl group, or a linear or branched saturated C7alkyl group, or a linear or branched saturated C8alkyl group, or a linear or branched saturated C9alkyl group, or a linear or branched saturated C10alkyl group, or a linear or branched saturated C11alkyl group, or a linear or branched saturated C12alkyl group, or a linear or branched saturated C13alkyl group, or a linear or branched saturated C14alkyl group, or a linear or branched saturated C15alkyl group.

In various embodiments, the linear or branched C1-C15alkyl group may be a linear or branched C2-C3alkenyl group, or a linear or branched C2-C4alkenyl group, or a linear or branched C2-C5alkenyl group, or a linear or branched C2-C6alkenyl group, or a linear or branched C2-C7alkenyl group, or a linear or branched C2-C8alkenyl group, or a linear or branched C2-C9alkenyl group, or a linear or branched C2-C10alkenyl group, or a linear or branched C2-C11alkenyl group, or a linear or branched C2-C12alkenyl group, or a linear or branched C2-C13alkenyl group, or a linear or branched C2-C14alkenyl group, or a linear or branched C2-C15alkenyl group, or a linear or branched C2alkenyl group, or a linear or branched C3alkenyl group, or a linear or branched C4alkenyl group, or a linear or branched C5alkenyl group, or a linear or branched C6alkenyl group, or a linear or branched C7alkenyl group, or a linear or branched C8alkenyl group, or a linear or branched C9alkenyl group, or a linear or branched C10alkenyl group, or a linear or branched C11alkenyl group, or a linear or branched C12alkenyl group, or a linear or branched C13alkenyl group, or a linear or branched C14alkenyl group, or a linear or branched C15alkenyl group.

In various embodiments, the linear or branched C1-C15alkyl group may be a linear or branched C2-C3alkynyl group, or a linear or branched C2-C4alkynyl group, or a linear or branched C2-C5alkynyl group, or a linear or branched C2-C6alkynyl group, or a linear or branched C2-C7alkynyl group, or a linear or branched C2-C8alkynyl group, or a linear or branched C2-C9alkynyl group, or a linear or branched C2-C10alkynyl group, or a linear or branched C2-C11alkynyl group, or a linear or branched C2-C12alkynyl group, or a linear or branched C2-C13alkynyl group, or a linear or branched C2-C14alkynyl group, or a linear or branched C2-C15alkynyl group, or a linear or branched C2alkynyl group, or a linear or branched C3alkynyl group, or a linear or branched C4alkynyl group, or a linear or branched C5alkynyl group, or a linear or branched C6alkynyl group, or a linear or branched C7alkynyl group, or a linear or branched C8alkynyl group, or a linear or branched C9alkynyl group, or a linear or branched C10alkynyl group, or a linear or branched C11alkynyl group, or a linear or branched C12alkynyl group, or a linear or branched C13alkynyl group, or a linear or branched C14alkynyl group, or a linear or branched C15alkynyl group.

In various embodiments, the C3-C18cycloalkyl group may be a C3-C4cycloalkyl group, or a C3-C5cycloalkyl group, or a C3-C6cycloalkyl group, or a C3-C7cycloalkyl group, or a C3-C8cycloalkyl group, or a C3-C9cycloalkyl group, or a C3-C10cycloalkyl group, or a C3-C11cycloalkyl group, or a C3-CL2cycloalkyl group, or a C3-C13cycloalkyl group, or a C3-C14cycloalkyl group, or a C3-C15cycloalkyl group, or a C3-C16cycloalkyl group, or a C3-C17cycloalkyl group, or a C3-C18cycloalkyl group, or a C4-C5cycloalkyl group, or a C4-C6cycloalkyl group, or a C4-C7cycloalkyl group, or a C4-C8cycloalkyl group, or a C4-C9cycloalkyl group, or a C4-C10cycloalkyl group, or a C4-C11cycloalkyl group, or a C4-C12cycloalkyl group, or a C4-C13cycloalkyl group, or a C4-C14cycloalkyl group, or a C4-C15cycloalkyl group, or a C4-C16cycloalkyl group, or a C4-C17cycloalkyl group, or a C4-C18cycloalkyl group, or a C3cycloalkyl group, or a C4cycloalkyl group, or a C5cycloalkyl group, or a C6cycloalkyl group, or a C7cycloalkyl group, or a C8cycloalkyl group, or a C9cycloalkyl group, or a C10cycloalkyl group, or a C11cycloalkyl group, or a C12cycloalkyl group, or a C13cycloalkyl group, or a C14cycloalkyl group, or a C15cycloalkyl group, or a C16cycloalkyl group, or a C17cycloalkyl group, or a C18cycloalkyl group.

In various embodiments, the C3-C18cycloalkyl group may be a C3-C4cycloalkenyl group, or a C3-C5cycloalkenyl group, or a C3-C6cycloalkenyl group, or a C3-C7cycloalkenyl group, or a C3-C8cycloalkenyl group, or a C3-C9cycloalkenyl group, or a C3-C10cycloalkenyl group, or a C3-C11cycloalkenyl group, or a C3-C12cycloalkenyl group, or a C3-C13cycloalkenyl group, or a C3-C14cycloalkenyl group, or a C3-C15cycloalkenyl group, or a C3-C16cycloalkenyl group, or a C3-C17cycloalkenyl group, or a C3-C18cycloalkenyl group, or a C4-C5cycloalkenyl group, or a C4-C6cycloalkenyl group, or a C4-C7cycloalkenyl group, or a C4-C8cycloalkenyl group, or a C4-C9cycloalkenyl group, or a C4-C10cycloalkenyl group, or a C4-C11cycloalkenyl group, or a C4-C12cycloalkenyl group, or a C4-C13cycloalkenyl group, or a C4-C14cycloalkenyl group, or a C4-C15cycloalkenyl group, or a C4-C16cycloalkenyl group, or a C4-C17cycloalkenyl group, or a C4-C18cycloalkenyl group, or a C3cycloalkenyl group, or a C4cycloalkenyl group, or a C5cycloalkenyl group, or a C6cycloalkenyl group, or a C7cycloalkenyl group, or a C8cycloalkenyl group, or a C9cycloalkenyl group, or a C10cycloalkenyl group, or a C11cycloalkenyl group, or a C12cycloalkenyl group, or a C13cycloalkenyl group, or a C14cycloalkenyl group, or a C15cycloalkenyl group, or a C16cycloalkenyl group, or a C17cycloalkenyl group, or a C18cycloalkenyl group.

In various embodiments, the C3-C18cycloalkyl group may be a C3-C4cycloalkynyl group, or a C3-C5cycloalkynyl group, or a C3-C6cycloalkynyl group, or a C3-C7cycloalkynyl group, or a C3-C8cycloalkynyl group, or a C3-C9cycloalkynyl group, or a C3-C10cycloalkynyl group, or a C3-C11cycloalkynyl group, or a C3-C12cycloalkynyl group, or a C3-C13cycloalkynyl group, or a C3-C14cycloalkynyl group, or a C3-C15cycloalkynyl group, or a C3-C16cycloalkynyl group, or a C3-C17cycloalkynyl group, or a C3-C18cycloalkynyl group, or a C4-C5cycloalkynyl group, or a C4-C6cycloalkynyl group, or a C4-C7cycloalkynyl group, or a C4-C8cycloalkynyl group, or a C4-C9cycloalkynyl group, or a C4-C10cycloalkynyl group, or a C4-C11cycloalkynyl group, or a C4-C12cycloalkynyl group, or a C4-C13cycloalkynyl group, or a C4-C14cycloalkynyl group, or a C4-C15cycloalkynyl group, or a C4-C16cycloalkynyl group, or a C4-C17cycloalkynyl group, or a C4-C18cycloalkynyl group, or a C3cycloalkynyl group, or a C4cycloalkynyl group, or a C5cycloalkynyl group, or a C6cycloalkynyl group, or a C7cycloalkynyl group, or a C8cycloalkynyl group, or a C9cycloalkynyl group, or a C10cycloalkynyl group, or a C11cycloalkynyl group, or a C12cycloalkynyl group, or a C13cycloalkynyl group, or a C14cycloalkynyl group, or a C15cycloalkynyl group, or a C16cycloalkynyl group, or a C17cycloalkynyl group, or a C18cycloalkynyl group.

In various embodiments, the linear or branched C1-C15alkyl group or the C3-C18cycloalkyl group may be intercepted with at least one heteroatom that may independently be O, S, N or P; or S, N or P; or O, N or P; or O, S, or P; or O, S, or N; or N or P; or S or P; or S or N; or O or P; or O or N; or O or S; or P; or N; or S; or O.

In various embodiment, the C1-C15alkyl group or the C3-C18cycloalkyl group may be substituted with one or more substituents selected from the group consisting of F, Cl, Br, I, —N+(C1-15alkyl)3, —N+(C2-15alkenyl)3, —N+(C2-15alkynyl)3, —N+H(C1-15alkyl)2, —N+H(C2-15alkenyl)2, —N H(C2-15alkynyl)2, P+(C1-15alkyl)3, P+(C2-15alkenyl)3, P+(C2-15alkynyl)3, S+(C1-15alkyl)2, S+(C2-15alkenyl)2, S+(C2-15alkynyl)2, oxo (i.e., ═O), —OC1-15alkyl, unsubstituted or substituted aromatic C3-C18cycloalkyl, unsubstituted or substituted non-aromatic C3-C18heterocyclic group, unsubstituted or substituted aromatic C3-C18heterocyclic group, ═NH, —C1-C15alkyl, non-aromatic C3-C18cycloalkyl, —N(C1-15alkyl)2, —N(C2-15alkenyl)2, —N(C2-15alkynyl)2, —COOH, —NH2, —SH, a biomolecule, and a linking group optionally joined to a biomolecule. In various embodiments, the C1-C15alkyl group or the C3-C18cycloalkyl group may be substituted with one or more substituents selected from the group consisting of F, Cl, Br, I, —N+(C1-15alkyl)3, —N+(C2-15alkenyl)3, —N+(C2-15alkynyl)3, —N+H(C1-15alkyl)2, —N+H(C2-15alkenyl)2, —NH(C2-15alkynyl)2, P+(C1-15alkyl)3, P+(C2-15alkenyl)3, P+(C2-15alkynyl)3, S+(C1-15alkyl)2, S+(C2-15alkenyl)2, S+(C2-15alkynyl)2, oxo (i.e., ═O), —OC1-15alkyl, unsubstituted or substituted aromatic C3-C18cycloalkyl, unsubstituted or substituted non-aromatic C3-C18heterocyclic group, unsubstituted or substituted aromatic C3-C18heterocyclic group, —C1-C15alkyl, non-aromatic C3-C18cycloalkyl, —N(C1-15alkyl)2, —N(C2-15alkenyl)2, —N(C2-15alkynyl)2, —COOH, —NH2, —SH, a biomolecule, and a linking group optionally joined to a biomolecule. In various embodiments, the C1-C15alkyl group or the C3-C18cycloalkyl group may be substituted with one or more substituents selected from the group consisting of a biomolecule and a linking group optionally joined to a biomolecule.

The compounds may also include base-free forms, prodrugs, or pharmaceutically acceptable salts thereof. The compounds described herein are meant to include all racemic mixtures and all individual enantiomers or combinations thereof, whether or not they are represented herein.

Various embodiments of the invention provide a method of making a precursor compound which comprises converting a corresponding alkylboronic acid variant of any of the aforementioned compounds to an alkylboronic ester, wherein one, two or three of Y1and Y2may be a leaving group displaceable by fluoride. This invention also provides a method of making a18F containing compound which comprises replacing at least one of said leaving group or groups of the aforesaid alkylboronic ester with18F. Methods of this invention make use of substituted alkyl-boronic compounds, wherein Y1or Y2may be a moiety that can be displaced by reaction with fluoride. These may be used as precursor molecules to the above-described18F labeled alkyfluoroborate compounds by reaction with a suitable source of18F. In this aspect of the invention, Y1or Y2may (for example) be an alkoxy, halide, amine (e.g. alkyl, aryl), or thiol (alkyl, aryl) moiety. Other examples of leaving groups that can be displaced by fluoride are known. Particular examples are described in WO 2005/077967.

Alkylfluoroborate compounds according to a formula described herein may be made by a variety of synthetic methods, ranging in complexity from de novo synthesis to a ‘wash-in’ of the fluorine on a previously prepared boronate (see, for example, Molander et al., J. Org. Chem. 2010, 75, 4304-4306; Lennox et al. JACS, 2012, 134, 7431-7441; Dumas et al., Organic Letters, 2012, 14, 2138-2141; and Molander et al., Organic Letters, 2006, 8, 2031-2034).

A ‘wash-in’ preparative method may include preparation of a solution of a substituted alkylboronic acid or ester in an appropriate solvent, to which aqueous fluoride is added. The pH may be at a suitable range (e.g. about 2.5 to about 3.5) or according to what is suitable for the solvents and the substituted alkylboronic acid or ester. The solvent may be DMF or another solvent that is miscible with an aqueous fluoride solution, and solubilizes the substituted alkylboronic acid or ester of interest. Examples of such solvents may include aqueous mixtures comprising DMSO, DMF, MeOH, THF, MeCN, DMA, and NMP. Selection of a particular solvent may vary with the particular substituted alkylboronic acid or ester and in particular with regards to preserving the bioactivity of the biomolecule, and is within the skill of one versed in the art. Aqueous fluoride may be at any suitable concentration. For example, the substituted alkylboronic acid may be present at a concentration of about 1-4 mM, and the KH18/19F2may be present in 3 or 4 equivalents i.e. 3-12 mM or 4-16 mM, where the minimal concentrations are selected to increase the specific activity of the labeling, and the maximal concentrations determined by the maximal solubility of the bioconjugate.

Clinical preparations of a substituted alkylboronic acid or ester may involve use of about 800 mCi18F in no carrier added form, which, barring environmental contamination with19F, represents about 0.46 nmol of18F, or 3.8% of the total fluoride used in a 10 μL reaction at 12 mM total fluoride. Microreactor and microfluidic techniques, (which provide reaction volumes of about 50 nL) can reduce the quantity of carrier19F needed. For instance in a 50 nL reaction at 10 mM fluoride, one needs only 500 pmol of total fluoride. Thus, a no carrier added reaction is readily contemplated for labeling alkylboronic acids.

Reaction temperature may be increased above room temperature, but below a temperature that may destabilize or denature the selected biomolecule. For example, some nucleic acids or oligonucleotides may be suitable for use in labeling reactions at temperatures of about 60° C., while some proteins may require lower temperatures. Peptides are known to withstand temperatures as high as about 110° C. Antibodies are known in the art to have limited thermostability compared to most other proteins however thermostable antibodies and enzymes may also be suitable for use in labeling reactions at temperatures above room temperature. Alternatively, some biomolecules may be preferentially suitable for labeling reactions at reduced temperatures, i.e. below room temperature.

General approaches and methods for direct chemical modification of biomolecules for addition and/or substitution of modifying groups are known. As an example, chemical modification of proteins is described by Means and FeeneyBioconjugate Chemistry1990 1: 2-12). Chemical modification of nucleic acids such as DNA and RNA is described in, for example, Boutourine et al.Bioconjugate Chemistry1990 1:350-56. Chemical modification of sugars and oligosaccharides is described in, for example Wood et al.Bioconjugate Chemistry1992 3: 391-6 and more recently in click reactions

In various embodiments, there is provided a method for selections or screening substituted organotrifluoroborate compounds, for their ability to resist defluoridation as an indicator of their longevity as a radio imaging agent for PET. Various methods may be employed such as described below.

Chromatographic methods for separation of a fluoridated compound from the free18F are useful for qualitative or semi-quantitative assessment of the resistance to defluoridation of the fluoridated compound. Such methods generally involve a stationary phase, which may be a column matrix having qualities such as hydrophobicity, porosity or size-exclusion capabilities, charge, hydrophilicity or the like. Alternately, the stationary phase may provide structural support only, and be largely inert to interactions with a mobile phase or the solutes in the mobile phase. The stationary phase may be further supported in, for example, a column, or for thin-layer chromatography, on a glass plate. Paper used in paper chromatography may provide both the stationary phase and physical support of the stationary phase. Alternatively a diol column can be used to remove unlabelled boronic acid that may be present as unreacted starting material or following competing solvolysis of the organotrifluoroborate during labeling. The mobile phase is frequently a solvent, which may be hydrophobic or hydrophilic, aqueous or non-aqueous, and may be formulated to provide a fixed pH or a selected pH range, or a particular salt or other solute concentration. In various embodiments, the molecules or compounds of interest, such as the fluoridated compounds above are soluble in the mobile phase, as is the free fluorine that is to be separated from the fluoridated compound. Choice of a particular chromatographic method may be influenced by the molecule to be separated. For example, separation of a labeled biomolecule, such as an antibody, gel-permeation or affinity chromatography may be suitable. In another example, separation of a labeled oligonucleotide or peptide, anion exchange chromatography may be suitable. In another example, separation of labeled biotin complexed with an avidin-conjugate may involve gel-permeation chromatography. In another example, separation of labeled free biotin, folate or methotrexate or other small molecules, such as peptides may involve chromatographic separation through a silica column or plug, or HPLC/FPLC.

Isotopic wash-out or pulse-chase methods may also be used for screening labeled trifluoroborates in order to identify compositions that are suitably stable for use as imaging agents.

It is recognized that stable organotrifluoroborates (no18F incorporated) may be useful as radiochemically stable precursors that will find use in labeling whereby they are treated under acidic conditions to promote the exchange of an atom of19F for an atom of18F. Following labeling, at least one of F may be18F. It may be recognized that in the preparation of compounds of this invention, that there may be a fraction of molecules that will not be complexed with any18F when carrier19F may be used. It may be recognized that the addition of carrier19F may be advantageous in certain cases. A final/overall specific activity suitable for imaging purposes can be achieved even if a particular trifluoroborate molecule in a mixture contains no18F, provided that at least some of the trifluoroborates prepared contain at least one18F.

EXAMPLES

Aryltrifluoroborates undergo solvolysis (i. e., B—F bond scission) at a nearly pH-independent rate above pH 3. The electronic nature of the ring substituents is the main factor affecting the rate of solvolysis. Groups on aryl rings impart their effects on rates by both inductive and resonance effects whereby the electron density in the pi-bonds is increased or diminished. Increased pi electron density can delocalize from the ring into the sp2 orbital on boron to promote loss of one fluoride ion whereupon the other two are lost rapidly. Increasing the electron density results in an enhancement of the rate of B—F bond scission, whereas decreasing the electron density reduces the rate. The inductive effects are difficult to separate from those related to resonance, and both effects are further nuanced by whether the substituents are in the meta vs ortho/para positions. Moreover, it is critical to understand the magnitude of these effects in order to understand how to create aryltrifluoroborates with sufficient stability.

The magnitude of these effects may be given by the ρ value in a quantitative linear free energy relationship. The effects of substitution by several (at least two) electron withdrawing group may be estimated by summing their known σ values. The rate at which such a composition reacts can be related to the rate of an unsubstituted aryl-BF3by the following equation (log(k/k0)=σρ where k is the solvolysis rate constant for a certain aryl-BF3under consideration and k0 is the solvolysis rate constant for an unsubstituted aryl-BF3Thus, for aryl-BF3solvolysis, the relationship between the rate of B—F bond scission and the number and position of various electron withdrawing groups may be understood in terms of known σ values and a value of ρ˜−1. Furthermore, owing to the change in the hybridization of boron from sp3 to sp2 upon loss of a first fluoride ion, which places the remaining two fluorine atoms co-planar with the aryl ring, it has been recognized that bulky ortho substituents may exert steric effects to retard solvolysis.

While aryl-trifluoroborate solvolysis can be understood in terms of delocalization of electron density from the arene pi-system into the empty p-orbital on boron with concomitant expulsion of a fluoride atom whose rate is correlated by the Hammett relationship of (log(k/k0)=σρ where ρ˜−1, this understanding based on a combination of inductive effects and pi-delocalization effects, as expressed in the pKa of (aryl) benzoic acids, and cannot be extended to substituted nonaromatic organotrifluoroborates. Electron withdrawing groups can greatly affect the rate of a reaction in some cases, but not in others, and thus the application of linear free energy relationships that hold well for aryl systems do not necessarily apply to nonaromatic compositions. It was unclear to what extent such relationships could be extended to the rate of B—F bond scission in systems where the carbon atom to which the boron is attached is not in an aromatic state, and of what magnitude these effects would be.

Fluoridation of a organoboronic Acid or Ester

22.4 mg (0.10 mmol) of N,N,-dimethyl-N-propargylammoniomethylborate pinacol ester was dissolved with 80 μL of DMF, then 2.4 equiv. 120 μL of 3 M KHF2(aqueous solution) and 80 μL of 4 M HCl (aqueous solution) were added as the fluoridation reagents. This reaction was incubated at 37° C. for two hours, then it was concentrated with a vacuum concentrator (Speedvac). A yellowish white powder was achieved and was ready for further purification.

Evaluation of Rates of Solvolytic Defluoridation of Fluoridated Organotrifluoroborates by 19F-Nuclear Magnetic Resonance (19F-NMR) Spectroscopy

The following is are representative examples of testing of solvolytic defluoridation of fluoridated organoborates as shown inFIG. 1AtoFIG. 1FFand Table 1. Solutions of the fluoridated organotrifluoroborates were made by dissolving 30 mg of the fluoridated organotrifluoroborate into 1 mL of 20% ethanol/acetonitrile.

As the fluoridated organotrifluoroborate salts were often contaminated with a significant amount of free fluoride, a silica column was used to remove the free fluoride from the organotrifluoroborate solution. After loading the crude reaction onto the silica column, the fractions containing the desired organotrifluoroborate were eluted and combined to form a combined solution. In view of the strong stability of the fluoridated organotrifluoroborate in organic solvent, 20% of the combined solution was stored in an NMR tube and this sample served as the “zero” minute time point. A purified organotrifluoroborate salt was isolated from the remaining portion of the combined solution (remaining 80% of the combined solution) by removing the organic solvent from the combined solution by vacuum. Removal of the organic solvent from the combined solution gave the purified fluoridated alkylbornate salt in the form of a white solid. The dry and purified fluoridated organotrifluoroborate salt was re-dissolved into 3 mL of 200 mM phosphate buffer. Solvolytic defluoridation started immediately upon dissolution of the fluoridated organotrifluoroborate salt into phosphate buffer, and accordingly, the time was recorded at the moment of dissolution. The progress of solvolytic defluoridation of the fluoridated organotrifluoroborate salt was monitored using19F NMR spectroscopy. General principles, methods and background relating to the use of19F NMR spectroscopy for monitoring defluoridation are known, and may be found in, for example, Ting et al. (2008, J. Org. Chem. 73; 4662-70; Harwig et al. (2008) Tetrahedron Letters 49:3152-56; and Ting et al. (2008) Journal of Fluorine Chemistry 129:349-58. For each time point, solvolytic defluoridation data was collected using a 300 MHz19F NMR spectrometer (Bruker).19F NMR spectroscopic tracers for the fluoridated organotrifluoroborate salt at different times are shown inFIG. 1A. Use of19F-NMR to measure solvolytic defluoridation provides a useful model for measuring solvolytic 18F-defluoridation because if one F is lost, all are lost. Moreover, it is understood that the kinetic isotope effect in terms of B—F bond solvolysis is negligible. Hence whereas for PET scanning purposes solvolytic 18F-defluoridation is of consideration,19F-NMR provides a reliable measure of the rate of 18F-defluoridation since both18F and19F rates are the same in practice because there should be no measurable isotopic difference. The degree of solvolytic defluoridation of the fluoridated organotrifluoroborate salt was plotted as a function of time, for example, as shown inFIG. 1Bfor Compound 6 of Table 1.19F NMR spectroscopic tracers for the fluoridated organotrifluoroborate salts of compounds listed in Table 1 at different times are shown inFIGS. 1A, 1C, 1E, 1G, 1I, 1K, 1M, 1O, 1Q, 1S, 1U, 1W, 1Y, 1AA, 1CC, and 1EE, and the degree of solvolytic defluoridation of the compounds as a function of time is shown inFIGS. 1B, 1D, 1F, 1H, 1J, 1L, 1N, 1P, 1R, 1T, 1V, 1X, 1ZZ, 1BB, 1DD, and 1FF. As can be seen fromFIGS. 1B, 1D, 1F, 1H, 1J, 1L, 1N, 1P, 1R, 1T, 1V, 1X, 1ZZ, 1BB, 1DD, and 1FF, solvolytic defluoridation of the fluoridated organoborate salts shows pseudo first-order reaction kinetics.

Rate Constant for Solvolytic Defluoridation

The time curves shown inFIGS. 1B and 1Dwere fitted and a rate constant, ksovolysis, for each time curve was calculated. As a measure of the stability of the fluoridated organoborate to solvolytic defluoridation, pkB—Fvalues were calculated using the equation pkB—F=−log(ksolvolysis). In addition to pkB—Fvalues, observed solvolytic defluoridation half-life, t1/2, values and rate constants, ksolvolysis, for a number of different fluoridated organoborates are listed in Table 1. As can be seen from Table 1, the larger the pkB—Fvalue, the more stable the fluoridated organoborate was observed to be to solvolytic defluoridation.

It has been reported that aryltrifluoroborates can be considerably stabilized by an electron-poor environment (see, for example, Ting et al. (2008, J. Org. Chem. 73; 4662-70). It has also been reported that EWGs (electron withdrawing group) on the benzene ring can significantly slow down hydrolysis, resulting in longer solvolytic half-lives. A similar correlation with such substituent effects is seen with regards to the nitration of benzene or the solvolysis of substituted benzyl tosylates. Without being bound by theory, a possible mechanistic reason for this effect was that pi-electrons in the aryl ring could delocalize into the empty p-orbital on boron thus promoting B—F bond dissociation. As such, a group that weakened this effect could result in greater B—F bond stability. However, in contrast to aryltrifluoroborates, for organotrifluoroborates, there may or may not be pi-overlap by an electron withdrawing group and hence there is no reason a priori to have thought that electron withdrawing groups would have any effects on the B—F bond stability or to what extent they might have retarded solvolysis.

The existence of any relationship between pkB-Fobserved for a particular alkyl moiety of the fluoridated organotrifluoroborate compound and the pKa of a carboxylic acid conjugated to the same alkyl moiety was also investigated. The pKa values for the carboxylic acids conjugated to the same alkyl moieties of the fluoridated organotrifluoroborates tabulated in Table 1 are also included in the same table. For example, for comparison purposes with pkB-Ffor Compound 1 of Table 1, the pKa of the carboxylic acid

is also listed in Table 1. pkB-Fvalues for a number of fluoridated organotrifluoroborates were plotted as a function of the pKa values for the corresponding carboxylic acids as shown inFIG. 2. Unexpectedly, a simple yet previously unknown relationship was observed between pKa and pkB-Fas seen inFIG. 2, i.e. that the log of the pseudo-first order solvolytic rate constant, log ksolv.(B—F), of RBF3is correlated to the pKa of the corresponding carboxylic acid RCOOH (R=0.99) with a slope of ˜−1.4. Without being bound by theory, one possible explanation for this relationship could involve consideration of the impact of various substituents on the electronic environment experienced by the carboxylic acid and in turn, the impact of the various substituent groups on the dissociability of the carboxylic acid proton. The observation of a relationship between the pKa and pkB-Fvalues also provides, in turn, an unexpected relationship between the pKa values and the stability or half-life of the fluoridated organotrifluoroborate compounds in terms of solvolytic defluoridation. A pKa of 2.85 or lower for the corresponding COOH may inform the design of organotrifluoroborate compositions with solvolytic half-lives of about 1000 min or longer. The box provides an approximate range that is indicative, although not determinative, of preferred utility.

This discovery now permits the stability of such organotrifluoroborate structures to be predicted, such that candidates for organoorganotrifluoroborates that will be useful in imaging may be selected and tested. A person skilled in the art of synthesis may consider the pKa of any carboxylic acid (with whatever modifications) by measurement or which is known (e.g. Jencks, Bordwell, CRC Handbook) or predicted by computer simulation, and use this knowledge to contemplate and design alkyltrifluoroborate structures that would have sufficiently long half-lives for use in the production of radiotracers.

Accordingly, a person skilled in the art may exploit this newly discovered relationship to identify candidate organotrifluoroborate compounds that are most likely to be useful as imaging agents. However, the invention is in no way limited to compounds that follow this relationship. Even though certain useful nonaromatic substituted oragnotrifluoroborate structures may deviate from this relationship when considered based solely on the pKa of a corresponding carboxylic acid, a person skilled in the art will understand that certain second order effects may dampen or accentuate the impact of various substituents on the electronic environment. Such deviations may easily be understood in terms of second-order effects related to steric interactions, to the presence of a second ionizable or charged group, or to other constraints that become clear in light of this relationship. Moreover, this relationship may guide the person skilled in the art when considering compounds that have no corresponding carboxylic acid.

This relationship also allows an electron withdrawing group to be placed as close to the boron as the carbon to which the boron is attached, whereas the geometry of an aryl ring requires that any electron withdrawing group be placed at least two carbon atoms away from the boron.

Evaluation of Stability of18F-Radiolabeled Fluoridated Organotrifluoroboratesby Thin Layer Chromatography (TLC)

The following is a representative example of testing of solvolytic defluoridation of a fluoridated organotrifluoroborateas shown inFIG. 3. 2 nmol Rhodamine-BF3was added into 100 μL plasma, and then incubated at 37° C. After 2 hours, 200 μL acetonitrile (ACN) was added to precipitate the protein from the solution. The crude was filtered with 40 μm filter paper, and then a reddish purple solution was achieved. 1 μL of this solution was loaded on the TLC plate. The TLC plate was developed by 15% methanol/DCM. The TLC chromatograms for an aryltrifluoroborate (Rhodamine-ArBF3), Rhodamine-PyrBF3, and a quaternary ammoniomethyltrifluoroborate (Rhodamine-Compound 10 of Table 1) are shown asFIG. 3.

In the TLC chromatograms, Rhodamine B was conjugated with the trifluoroborate to trace the decomposition. The hydrolyzed product lost the negative charge, and therefore migrated much more rapidly than the precursor. Based on the TLC chromatograms the ArBF3showed decomposition after incubation in plasma for two hours. PyrBF3exhibited greater stability than ArBF3because less decomposition was detected on the TLC chromatogram. Moreover, for Compound 10 of Table 1, almost no hydrolyzed product was detected based on the fluorescence analysis. The lack of detection of hydrolyzed product for Compound 10 of Table 1 suggests that Compound 10 has extraordinary stability in plasma and may also show excellent stability in animal models.

Evaluation of Stability of18F-radiolabeled Fluoridated Organotrifluoroborates by High-performance Liquid Chromatography (HPLC)

1.1 mCi of18F-Compound 10 (18F-ammoniomethyltrifluoroborate) was mixed with 100 L of plasma to form a18F-Compound 10-plasma mixture, and then incubated at 37° C. for two hours. 200 uL of acetonitrile (ACN) was then added to the18F-Compound 10-plasma mixture to precipitate the protein from the mixture. The precipitated protein portion was separated from the mixture by filtration with 40 m filter paper, and the clear filtrate was injected into an HPLC for stability analysis. Chromatograms for independent HPLC tests of Rhodamine-Compound 10 of Table 1 at 0 minutes (A), 80 minutes (B), 150 minutes (C), 0 minutes (D), 150 minutes (E), and 150 minutes (F) are shown inFIG. 4. Based on the in vitro test (n=3), almost no decomposition was observed in 150 mins, which proved that Compound 10 has excellent stability in plasma. The observed excellent stability of Compound 10 in plasma suggest that it should present good stability under in vivo conditions.

Radiolabeling and Animal Study with Rhodamine(B)-AMBF3.

The following compound, wherein Rhodamine(B) is conjugated to Compound 10, was synthesized and injected into mice.

Compound 18 was synthesized according to the method outlined below:

FIG. 5Ashows HPLC traces of radioactivity in the plasma of a rat injected with Compound 18 at 0, 80 and 150 min post injection.FIG. 5Bshows PET/CT images of the mouse at 10, 30, and 60 min post injection.

Radiolabeling and Animal Study with Bissulfo-Rhodamine(B)-AMBF3.

The following compound, wherein sulforhodamine(B) is conjugated to Compound 13, was synthesized and injected into mice.

FIG. 6Ashow the uptake value of different organs for Rhodamine-Compound 10 at 60 min post injection into a mouse.FIG. 6Bis a PET/CT image of a mouse injected with Rhodamine-Compound 10 at 60 min post injection.

The following compound, Compound 10 conjugated to folic acid, has been synthesized.

Compound 20 was synthesized according to the method outlined below:

Various embodiments of the invention provide peptides or neurotracers conjugated to the positron-emitting organofluoroborate compounds described herein. A person skilled in the art will understand that the positron-emitting organofluoroborate compounds described herein can be conjugated to a ligand (e.g. a peptide or a neurotracer) with affinity and specificity for a biomolecule of interest, provided that a suitable linker is chosen such that the affinity of the ligand for the biomolecule of interest is not reduced by not more than a factor of 100. Such compounds may serve as useful tracers.

For example, BisRGD-rho-AMBF3(Compound 21)_has been synthesized according to the methods described in Zhibo Liu et al.MedChemComm2014 5: 171-179 and Liu et al.Nucl. Med and Biology2013 40: 841-849. Methods for conjugation may include use of an alkyne- or azide-linked tri-substituted ammonium methyl-BF3 or disubstituted protonated ammonium-methyl-BF3 that is linked to the peptide by copper-catalyzed or strain promoted cycloaddition reactions. A person skilled in the art will understand that rhodamine is used for the purposes of screening various RBF3 on TLC and for easily measuring the specific activity of a tracer, and has also been used specifically to direct tracers for cardiac imaging. However, those skilled in the art will further understand that the rhodamine can be replaced with a peptide or other ligand to be imaged.

FIG. 7shows PET-CT images of mice injected with Compound 21.

Although various embodiments of the invention are disclosed herein, many adaptations and modifications may be made within the scope of the invention in accordance with the common general knowledge of those skilled in this art. Such modifications include the substitution of known equivalents for any aspect of the invention in order to achieve the same result in substantially the same way. Numeric ranges are inclusive of the numbers defining the range. The word ‘comprising’ may be used herein as an open-ended term, substantially equivalent to the phrase ‘including, but not limited to’, and the word ‘comprises’ has a corresponding meaning. As used herein, the singular forms ‘a’, ‘an’ and ‘the’ include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to ‘a thing’ includes more than one such thing. Citation of references herein may be not an admission that such references are prior art to the present invention. Publications, including patents and patent applications, cited in this specification are incorporated herein by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein and as though fully set forth herein. The invention includes all embodiments and variations substantially as hereinbefore described and with reference to the examples and drawings.