A chemoenzymatic platform for the syntheses of clickable kainoid neurochemicals

Abstract

Presented are kainoids, kainoid precursors, and kainoid derivatives and synthetic schemes for producing kainoids, kainoid precursors, and kainoid derivatives. The synthetic schemes for producing kainoids can make use of kainoid synthases, for example, in cyclizing a kainoid precursor to produce a kainoid. The kainoids can include an alkyne group 5 to facilitate the production of kainoid derivatives.

Description

[0001] A CHEMOENZYMATIC PLATFORM FOR THE SYNTHESES OF CLICKABLE KAINOID NEUROCHEMICALS

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 708,251 filed October 16. 2024.

[0003] FIELD OF THE EMBODIMENTS

[0004] This specification pertains to the field of kanoids and their synthesis.

[0005] SUMMARY

[0006] Some embodiments of the invention include the following:

[0001] A compound of formula wherein n is at least 1.

[0007] [2] The compound of [1], wherein n is 1 or 2.

[0008] [3] The compound of | 1 ], of formula

[0009] [4] The compound of [1], of formula

[0010]

[0011] [5] A compound of formula wherein m is at least 0.

[0012] [6] The compound of [5], wherein m is 0 or 1.

[0013] [7] The compound of [5] of formula

[0014] [8] The compound of [5] of formula

[0015]

[0016] [9] A compound of formula wherein m is at least 0 and wherein G is is selected from the group consisting of a fluorescent dye, a coumarin dye, coumarin, a tri aryl methane dye, fluorescein, a cyanme dye, Cy5, a squaraine dye, SeTau-647, biotin, and benzyl.

[0010] The compound of [9], wherein m is 0 or 1

[0017]

[0011] The compound of [9] of formula

[0012] The compound of [9] of formula

[0018]

[0013] The compound of claim [9] of formula

[0019]

[0014] The compound of [9] of formula

[0020]

[0015] A method of preparing a compound of formula reacting a 1stcompound

[0021] (1stcompound) with an epoxidizing agent to obtain a 2ndcompound

[0022] (2ndcompound) reacting the 2ndcompound with an oxidizing agent to obtain a 3rdcompound

[0023] (3rdcompound) reacting the 3rdcompound with a homologation agent, optionally a Seyferth-Gilbert reagent, dimethyl (diazomethyl)phosphonate, and / or dimethyl 1 -diazo-2-oxopropylphosphonate, to obtain a 4thcompound

[0024] (4thcompound) reacting the 4thcompound with an oxidizing agent to obtain a 5thcompound

[0025] (5thcompound) reacting the 5thcompound with a glutamic acid (Glu), wherein n is at least 1 .

[0026]

[0016] The method of

[0015] , wherein n is 1 or 2.

[0027]

[0017] The method of

[0015] , comprising preparing a compound of formula

[0028] (an alkynyl-functionalized prekainic acid) comprising reacting a 1stcompound with meta-chloroperoxybenzoic acid (mCPBA) in dichloromethane (DCM) to obtain a 2ndcompound reacting the 2ndcompound with an alkali metal periodate in tetrahydrofuran (THF) and water (H2O) to obtain a 3rdcompound (3rdcompound) reacting the 3rdcompound with (compound A) and an alkali metal carbonate in methanol (MeOH) to obtain a 4thcompound (4thcompound) reacting the 4thcompound with Dess-Martin periodinane (DMP) in dichloromethane (DCM) to obtain a 5thcompound (5thcompound) reacting the 5thcompound with L-glutamic acid (L-Glu) in a solution of an alkali metal borohydride and an alkali metal hydroxide in methanol (MeOH) and water (H2O) to obtain the alkynyl-functionalized prekainic acid.

[0018] The method of

[0017] , wherein the alkali metal periodate is sodium periodate (NaIO4), wherein the alkali metal carbonate is potassium carbonate (K2CO3), wherein the alkali metal borohydride is sodium borohydride (NaBH4), and wherein the alkali metal hydroxide is sodium hydroxide (NaOH).

[0019] A method of preparing a compound of formula comprising transforming with a kanoid synthase, wherein m is at least 0.

[0029]

[0020] The method of

[0019] , wherein m is 0 or 1.

[0030]

[0021] The method of

[0019] , wherein the kanoid synthase has at least 80%, 90%, 95%, 98%, or 99%, amino acid sequence identify to a KabC synthase, a DabC synthase, a RadCl synthase, DsKabC [SEQ ID NO. 1], GfKabC [SEQ ID NO. 2], ReKabC [SEQ ID NO. 3], PpKabC [SEQ ID NO. 4], PmDabC [SEQ ID NO. 5], or CaRadCl [SEQ ID NO. 6].

[0031]

[0022] The method of

[0019] , wherein the kanoid synthase is selected from the group consisting of a KabC synthase, aDabC synthase, and aRadCl synthase.

[0032]

[0023] The method of

[0019] , wherein the kanoid synthase is selected from the group consisting of DsKabC [SEQ ID NO. 1], GfKabC [SEQ ID NO. 2], ReKabC [SEQ ID NO. 3], PpKabC [SEQ ID NO. 4], PmDabC [SEQ ID NO. 5], and CaRadCl [SEQ ID NO. 6],

[0033]

[0024] The method of

[0019] , comprising preparing a compound of formula

[0034]

[0035] (an alkynyl-functionalized kainic acid) comprising transforming

[0036] (an alkynyl-functionalized prekainic acid) with CuRadC 1 [SEQ ID NO. 6] protein in a solution of a-ketoglutaric acid (aKG), iron(ll) sulfate

[0037] (FeSO4), and L-ascorbic acid (L-asc) in water (H2O) to obtain the alkynyl-functionalized kainic acid.

[0038]

[0025] A method of preparing a compound of formula comprising reacting

[0039] with an azide-functionalized group G, optionally in the presence of a copper salt, wherein m is al least 0.

[0026] The method of

[0025] . wherein m is 0 or I

[0040]

[0027] The method of

[0025] , comprising preparing a compound of formula

[0041] (group G-functionalized triazole-functionalized kainic acid) comprising

[0042] (an alkynyl-functionalized kainic acid) with an azide-functionalized group G, optionally in the presence of a copper salt, to obtain the group G-functionalized triazole-functionalized kainic acid.

[0028] The method of any one of claims [251 -

[0027] , wherein the group G is selected from the group consisting of a fluorescent dye, a coumarin dye, coumarin, a triarylmethane dye, fluorescein, a cyanine dye, Cy5, a squaraine dye. SeTau-647, biotin, and benzyl.

[0043]

[0029] The method of any one of claims

[0025] -

[0028] , wherein the copper salt is copper(II) sulfate (CuSO4).

[0044]

[0030] The method of any one of claims

[0025] -

[0029] , wherein the reacting is further in the presence of an alkali ascorbate and'or sodium ascorbate.

[0045]

[0031] The method of any one of claims

[0025] and

[0027] -

[0030] , comprising preparing a compound of formula

[0046] (coumann-triazole functionalized kainic acid) comprising

[0047] (an alkynyl-functionalized kainic acid) with azide-functionalized coumarin (coumarin-N3) in the presence of a copper salt to obtain the coumarin- triazole functionalized kainic acid

[0032] The method of any one of claims

[0025] and

[0027] -

[0030] , comprising preparing a compound of formula (benzyl-triazole functionalized kainic acid) comprising reacting (an alkynyl-functionalized kainic acid) with benzyl azide (Bn-N3) in the presence of a copper salt to obtain the benzyl-triazole functionalized kainic acid. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 illustrates synthetic strategies for generating C1’ functionalized kainoid neurochemicals from 4-hydroxy-L-proline precursors. FIG. 2A shows a first route for the biosynthesis of native kainoids in marine micro- and macroalgae. FIG.2B shows a second route for the biosynthesis of native kainoids in marine micro- and macroalgae. FIG. 3A contrasts a synthetic approach using the enzyme KabC (A) with a synthetic approach using E. coli including the recombinant gene p28DsKabC. FIG.3B illustrates how E. coli is transformed with the recombinant gene p28DsKabC and is treated to obtain the DsKabC [SEQ ID NO.1] enzyme in lysate. FIG.3C presents a synthesis using the DsKabC [SEQ ID NO.1] enzyme in lysate. FIG.3D presents a graph showing the conversion of prekainic acid to kainic acid over time in the synthesis of FIG.3C. FIG. 4 illustrates a retro(bio)synthetic analysis towards constructing terminal alkyne- containing kainoids using kainoid synthase enzymology and chemical synthesis from glutamate and terpenoid precursors. FIG. 5A shows a chemical synthesis of linear pre-alkynic (n = 1) and pre-aldynic (n = 2) acid substrates. FIG. 5B shows a trans-selective chemical synthesis of linear pre-alkynic (n = 1) and pre- aldynic (n = 2) acid substrates. FIG.6A shows a chemical synthesis with R selected from various groups shown in FIGS. 6B and 6C. FIG. 6B shows an in vitro lysate assay with six kainoid synthases against non-native cis / trans mixtures of pre-alkynic (2) and pre-aldynic (3) substrates (top) and native N-prenylated L-glutamate substrates (4-6) (bottom). A heatmap for cyclization activity is shown. FIG. 6C shows an in vitro lysate assay with six kainoid synthases against non-native cis / trans mixtures of pre-alkynic (2) and pre-aldynic (3) substrates (top) and native N-prenylated L-glutamate substrates (4-6) (bottom). A heatmap for hydroxylation activity is shown. FIG.6D shows a chemical synthesis with the CaRadC1 [SEQ ID NO.6] kanoid synthase. FIG. 6E shows UHPLC-MS analytical assays of CaRadC1 [SEQ ID NO. 6] transformations under the chemical synthesis of FIG.6D of pre-alkynic acid (2) cis / trans mixtures using optimized conditions (Figures 6F-6H). FIG. 6F shows a chemical synthesis with prealkynic acid (2) and the CaRadC1 [SEQ ID NO.6] kanoid synthase. FIG.6G shows extracted ion chromatograms (EICs) for the chemical synthesis of FIG.6F. FIG.6H shows a summary of results of the chemical synthesis of FIG.6F. FIG.6I shows a chemical synthesis with prealkynic acid (2) and a RadC1 or a DabC kanoid synthase. FIG.6J shows extracted ion chromatograms for the chemical synthesis of FIG.6I. FIG.6K shows a chemical synthesis with prealdynic acid (3) and the CaRadC1 [SEQ ID NO.6] kanoid synthase. FIG. 6L shows UHPLC-MS analytical assays of CaRadC1 [SEQ ID NO. 6] transformations under the chemical synthesis of FIG.6K of pre-aldynic acid (3) cis / trans mixtures using optimized conditions (Figures 6M-6O). FIG.6M shows a chemical synthesis with prealdynic acid (3) and the CaRadC1 [SEQ ID NO.6] kanoid synthase. FIG.6N shows extracted ion chromatograms for the chemical synthesis of FIG.6M. FIG.6O shows a summary of results of the chemical synthesis of FIG.6M. FIG.7A shows a chemical synthesis with prealkynic acid (2) and a kanoid synthase. FIG.7B shows extracted ion chromatograms for the chemical synthesis of FIG.7A. FIG. 7C shows a chemical synthesis with prealkynic acid (2) and the CaRadC1 [SEQ ID NO.6] kanoid synthase. FIG.7D shows extracted ion chromatograms for the chemical synthesis of FIG.7C. FIG. 7E shows an NMR spectrum for an isolated product of the CaRadC1 [SEQ ID NO. 6] transformation of prealkynic acid B (2). FIG. 7F shows an NMR spectrum for an isolated product of the CaRadC1 [SEQ ID NO. 6] transformation of prealkynic acid B (2). FIG. 7G shows an NMR spectrum for an isolated product of the CaRadC1 [SEQ ID NO. 6] transformation of prealkynic acid B (2). FIG. 8A shows reaction schemes starting with prealkynic acid (2) (top) and starting with prealdynic acid A (3) (bottom). Preparative enzymatic transformation of prealkynic acid (2) yields alkynic acid B (7); subsequent CuAAC functionalization with benzyl azide is shown (top). Preparative enzymatic transformation of prealdynic acid (3) yields aldynic acid A (8); subsequent CuAAC functionalization with benzyl azide is shown (bottom). Critical Nuclear Overhauser Effects (NOEs) determining C1’ stereochemistry are depicted with arrows. FIG.8B shows a click reaction scheme starting with alkynic acid B (7) and adding a benzyl group by reaction of the alkynic acid B (7) with benzyl (Bn) azide to result in benzyl-functionalized kainic acid (top) and extracted ion chromatograms for the reactant and the product (bottom). FIG. 8C shows a click reaction scheme starting with alkynic acid B (7) and adding a coumarin group by reaction of the alkynic acid B (7) with coumarin azide to result in comarin- functionalized kainic acid (top) and extracted ion chromatograms for the reactant and the product (bottom). The coumarin group can have fluorescent properties. FIG. 8D shows a reaction scheme starting with prealdynic acid A (3) (top) and extracted ion chromatograms (bottom). FIG.9A shows the peak response of glutamate, compound 1, compound 9, compound 10, compound 7, or compound 8 at recombinant homomeric GluK2 expressed in HEKT-293 cells and measured via outside-out patch clamp electrophysiology. Partial agonistic activity of non-native kainoids alkynic acid B 7 and aldynic acid B 8 is retained but weakened compared to native kainoids. FIG.9B shows the relative peak response of kainoids tested at 1 mM normalized to 10 mM glutamate. FIG.9C shows the current rise-time kinetic parameter of kainoid agonism. FIG.9D shows the desensitization decay kinetic parameter of kainoid agonism. FIG.9E shows the current equilibrium kinetic parameter of kainoid agonism. FIG. 10A illustrates glutamate bound (molecular docked) to a GluK2 ligand binding domain (LBD) (PDB ID 2XXR). FIG.10B illustrates kainic acid (1) bound to a GluK2 LBD (PDB ID 2XXT). FIG. 10C illustrates domoic acid bound to the full homomeric GluK2 channels (PDB ID 8GC2). FIG.10D illustrates the non-native kainoid alkynic acid B (7) docked into a GluK2 LBD via DynamicBind.24FIG.10E illustrates the non-native kainoid aldynic acid A (8) docked into a GluK2 LBD. FIG.10F illustrates domoic acid bound to a GluK1 LBD (PDB ID 2pbw). FIG.10G illustrates alkynic acid B (7) docked to a GluK1 LBD (PDB ID 2pbw). FIG.11A shows a proton (1H) nuclear magnetic resonance (NMR) spectrum taken at 500 MHz in deuterium oxide (D2O) + 0.1% methanol (MeOH) of kainic acid (1). FIG.11B shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of kainic acid (1). FIG.11C shows a heteronuclear single quantum correlation (HSQC) NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of kainic acid (1). FIG.12A shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of alkynic acid B (7). FIG.12B shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of alkynic acid B (7). FIG.12C shows a correlation spectroscopy (COSY) NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of alkynic acid B (7). FIG. 12D shows a HSQC NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of alkynic acid B (7). FIG. 12E shows a HSQC NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of alkynic acid B (7). FIG.12F shows a nuclear Overhauser effect spectroscopy (NOESY) NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of alkynic acid B (7). FIG.13A shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of aldynic acid A (8). FIG.13B shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of aldynic acid A (8). FIG. 13C shows a carbon 13 (13C) NMR spectrum taken at 125 MHz with deuterated methanol (MeOD) of aldynic acid A (8). FIG.13D shows a COSY NMR spectrum taken at 500 MHz with MeOD of aldynic acid A (8). FIG.13E shows a HSQC NMR spectrum taken at 500 MHz with MeOD of aldynic acid A (8). FIG. 13F shows a heteronuclear multiple-bond correlation spectroscopy (HMBC) NMR spectrum taken at 500 MHz with MeOD of aldynic acid A (8). FIG.13G shows a NOESY NMR spectrum taken at 500 MHz with MeOD of aldynic acid A (8). FIG.14A shows a1H NMR spectrum taken at 500 MHz with MeOD of dainic acid (9). FIG.14B shows a1H NMR spectrum taken at 500 MHz with MeOD of dainic acid (9). FIG.14C shows a HSQC NMR spectrum taken at 500 MHz with MeOD of dainic acid (9). FIG. 15A shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of isodomoic acid (10). FIG. 15B shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of isodomoic acid (10). FIG. 15C shows a COSY NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of isodomoic acid (10). FIG. 15D shows a HSQC NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of isodomoic acid (10). FIG.16 shows a1H NMR spectrum taken at 500 MHz in deuterated chloroform (CDCl3) of (E)-5-(3,3-dimethyloxiran-2-yl)-3-methylpent-2-en-1-yl acetate (S1). FIG.17 shows a1H NMR spectrum taken at 500 MHz in CDCl3of (E)-3-methyl-6-oxohex- 2-en-1-yl acetate (S2). FIG. 18 shows a1H NMR spectrum taken at 500 MHz in CDCl3of (E)-3-methylhept-2- en-6-yn-1-ol (S3). FIG.19A shows a1H NMR spectrum taken at 500 MHz in CDCl3of (E)-3-methylhept-2- en-6-ynal (S4). FIG.19B shows a two-dimensional (2D) NMR spectrum of (E)-3-methylhept-2-en-6-ynal (S4). FIG.19C shows a two-dimensional (2D) NMR spectrum of (E)-3-methylhept-2-en-6-ynal (S4). FIG. 20A shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealkynic acid (3-methylhept-2-en-6-yn-1-yl)-L-glutamic acid) (2). FIG. 20B shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealkynic acid (3-methylhept-2-en-6-yn-1-yl)-L-glutamic acid) (2). FIG. 20C shows a13C NMR spectrum taken at 125 MHz in D2O + 0.1% MeOH of prealkynic acid (3-methylhept-2-en-6-yn-1-yl)-L-glutamic acid) (2). FIG. 20D shows a COSY NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealkynic acid (3-methylhept-2-en-6-yn-1-yl)-L-glutamic acid) (2). FIG. 20E shows a HSQC NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealkynic acid (3-methylhept-2-en-6-yn-1-yl)-L-glutamic acid) (2). FIG. 20F shows a HMBC NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealkynic acid (3-methylhept-2-en-6-yn-1-yl)-L-glutamic acid) (2). FIG. 20G shows a NOESY NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealkynic acid (3-methylhept-2-en-6-yn-1-yl)-L-glutamic acid) (2). FIG.21 shows a1H NMR spectrum taken at 500 MHz in CDCl3of a farnesyl epoxide mix (S5 / S5b). FIG.22 shows a1H NMR spectrum taken at 500 MHz in CDCl3of (2E,6E)-3,7-dimethyl- 10-oxodeca-2,6-dien-1-yl acetate (S6). FIG. 23 shows a1H NMR spectrum taken at 500 MHz in CDCl3of (2E,6E)-3,7- dimethylundeca-2,6-dien-10-yn-1-ol (S7). FIG. 24A shows a1H NMR spectrum taken at 500 MHz in CDCl3of (2E,6E)-3,7- dimethylundeca-2,6-dien-10-ynal (S8). FIG. 24B shows a COSY NMR spectrum taken at 500 MHz in CDCl3of (2E,6E)-3,7- dimethylundeca-2,6-dien-10-ynal (S8). FIG. 24C shows a HSQC NMR spectrum taken at 500 MHz in CDCl3of (2E,6E)-3,7- dimethylundeca-2,6-dien-10-ynal (S8). FIG. 24D shows a HMBC NMR spectrum taken at 500 MHz in CDCl3of (2E,6E)-3,7- dimethylundeca-2,6-dien-10-ynal (S8). FIG. 24E shows a NOESY NMR spectrum taken at 500 MHz in CDCl3of (2E,6E)-3,7- dimethylundeca-2,6-dien-10-ynal (S8). FIG. 25A shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealdynic acid ((6E)-3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamic acid) (3). FIG. 25B shows a13C NMR spectrum taken at 125 MHz in D2O + 0.1% MeOH of prealdynic acid ((6E)-3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamic acid) (3). FIG. 25C shows a COSY NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealdynic acid ((6E)-3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamic acid) (3). FIG. 25D shows a HSQC NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealdynic acid ((6E)-3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamic acid) (3). FIG. 25E shows a HMBC NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealdynic acid ((6E)-3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamic acid) (3). FIG. 25F shows a NOESY NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of prealdynic acid ((6E)-3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamic acid) (3). FIG. 26A shows a1H NMR spectrum taken at 500 MHz in CDCl3of (E)-7-bromo-5- methylhept-5-en-1-yne (S9). FIG. 26B shows a1H NMR spectrum taken at 500 MHz in CDCl3of (E)-7-bromo-5- methylhept-5-en-1-yne (S9). FIG.26C shows a HSQC NMR spectrum taken at 500 MHz in CDCl3of (E)-7-bromo-5- methylhept-5-en-1-yne (S9). FIG.26D shows a HSQC NMR spectrum taken at 500 MHz in CDCl3of (E)-7-bromo-5- methylhept-5-en-1-yne (S9). FIG.26E shows a HMBC NMR spectrum taken at 500 MHz in CDCl3of (E)-7-bromo-5- methylhept-5-en-1-yne (S9). FIG.27A shows a1H NMR spectrum taken at 500 MHz in CDCl3of (di-tert-butyl (E)-(3- methylhept-2-en-6-yn-1-yl)-L-glutamate (S10). FIG.27B shows a13C NMR spectrum taken at 125 MHz in CDCl3of (di-tert-butyl (E)-(3- methylhept-2-en-6-yn-1-yl)-L-glutamate (S10). FIG.27C shows a COSY NMR spectrum taken at 500 MHz in CDCl3of (di-tert-butyl (E)- (3-methylhept-2-en-6-yn-1-yl)-L-glutamate (S10). FIG.27D shows a HSQC NMR spectrum taken at 500 MHz in CDCl3of (di-tert-butyl (E)- (3-methylhept-2-en-6-yn-1-yl)-L-glutamate (S10). FIG.27E shows a HMBC NMR spectrum taken at 500 MHz in CDCl3of (di-tert-butyl (E)- (3-methylhept-2-en-6-yn-1-yl)-L-glutamate (S10). FIG. 27F shows a NOESY NMR spectrum taken at 500 MHz in CDCl3of (di-tert-butyl (E)-(3-methylhept-2-en-6-yn-1-yl)-L-glutamate (S10). FIG.28A shows a1H NMR spectrum taken at 500 MHz in CDCl3of (E)-(3-methylhept-2- en-6-yn-1-yl)-L-glutamic acid (S11). FIG. 28B shows a COSY NMR spectrum taken at 500 MHz in CDCl3of (E)-(3- methylhept-2-en-6-yn-1-yl)-L-glutamic acid (S11). FIG. 28C shows a HSQC NMR spectrum taken at 500 MHz in CDCl3of (E)-(3- methylhept-2-en-6-yn-1-yl)-L-glutamic acid (S11). FIG. 29 shows a1H NMR spectrum taken at 500 MHz in CDCl3of (E)-(3-methylhept-2- en-6-yn-1-yl)-L-glutamic acid (S12). FIG.30A shows a1H NMR spectrum taken at 500 MHz in CDCl3of di-tert-butyl (2E,6E)- 3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamate (S13). FIG.30B shows a13C NMR spectrum taken at 125 MHz in CDCl3of di-tert-butyl (2E,6E)- 3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamate (S13). FIG. 30C shows a COSY NMR spectrum taken at 500 MHz in CDCl3of di-tert-butyl (2E,6E)-3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamate (S13). FIG. 30D shows a HSQC NMR spectrum taken at 500 MHz in CDCl3of di-tert-butyl (2E,6E)-3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamate (S13). FIG. 30E shows a HMBC NMR spectrum taken at 500 MHz in CDCl3of di-tert-butyl (2E,6E)-3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamate (S13). FIG. 30F shows a NOESY NMR spectrum taken at 500 MHz in CDCl3of di-tert-butyl (2E,6E)-3,7-dimethylundeca-2,6-dien-10-yn-1-yl)-L-glutamate (S13). FIG.31A shows a 1D NMR spectrum of alkynic acid benzyl triazole (11). FIG.31B shows a1H NMR spectrum taken at 500 MHz in D2O + 0.1% MeOH of alkynic acid benzyl triazole (11). DETAILED DESCRIPTION Kainate receptors (KARs) are a class of ionotropic glutamate receptors (iGluRs) that mediate excitatory neuronal signaling, modulate synaptic plasticity, and are implicated in multiple human psychiatric and neurological disorders ranging from epilepsy, autism, and Parkinson’s disease to schizophrenia, pain, and alcohol use disorder.1–3KARs are tetrameric ion channels composed of variable combinations of five subunits (GluK1-5), with the pharmacological and biophysical properties of full channels varying by subunit constitution.4While the armamentarium developed to study NMDA and AMPA receptors is relatively rich, fewer small molecules designed specifically for KARs and their subunits are available within a neuropharmacological toolbox.5Nonetheless, advances in KAR neurobiology and biophysics continue to make strides today and the development of efficacious KAR-selective and subtype specific pharmacologics is of intensifying focus to further accelerate our understanding of these receptors in mammalian physiology. KARs are named as such due to their high affinity for kainoid marine neurotoxins: a class of alkaloid meroterpenoids bearing a pyrrolidine ring that rigidifies a glutamic acid moiety in an entropically favorable orientation to outcompete endogenous L-glutamate within the KAR ligand binding domains (LBDs). Due to their excitatory potency, kainoids like kainic acid (1) have been used in neuropharmacology assays and epilepsy disease models.6That is, kainic acid and other members of the kainoid family of compounds, are neuropharmacology agents used in studying the role of ionotropic glutamate receptors, including the kainate receptor, in the nervous system, including the central nervous system. Kainic acid functions as an ionotropic glutamate receptor (iGluR) agonist. iGluRs mediate neuronal cell-cell commutation by binding to glutamate and facilitating influx of Ca2+into the cell. Structurally similar to glutamic acid, kainic acid can bind more efficiently to iGluRs and thereby stimulate excessive influx of Ca2+, which can lead to excitotoxicity and cell death. Kainic acid might not produce excitotoxic effects for humans in vivo; however, kainic acid's bioactivity has been exploited to create mouse model systems to study neurological diseases, for example, temporal lobe epilepsy. Kainic acid can be isolated from the seaweeds Digenea simplex (Ds) and Chondria armata (Ca). However, shortages of kainic acid have resulted in it having a large cost. Synthetic routes for obtaining kainic acid and other members of the kainoid family of compounds are useful. To supply the use of kainoids, synthetic routes have been developed ranging from Baldwin’s early synthesis of 1 from D-serine via cobalt-mediated radical cyclization, to Tian’s more diversifiable approaches from 4-hydroxy-L-proline (Figure 1).7,8The chemical approaches have enabled synthetic methodologies; however, kainoid biosynthesis by marine macro- and microalgae may be comparatively simple. Whereas chemical construction of the three contiguous stereocenters along the pyrrolidine core required multiple synthetic steps, biosynthetic carbon-carbon bond formation mediated by kainoid synthase enzymes completes this transformation in a single step (Figures 2A and 2B). For the synthesis of kainic acid from L-glutamic acid, a synthetic approach using the enzyme KabC (A) is contrasted with a synthetic approach using E. coli including the recombinant gene p28DsKabC that expresses the DsKabC enzyme [SEQ ID NO.1] (B) in Figure 3A. Figure 3B illustrates how E. coli is transformed with the recombinant gene p28DsKabC and is treated to obtain the DsKabC [SEQ ID NO. 1] (Digenea simplex (Ds) prekainate cyclase (KabC)) enzyme in lysate. Figure 3C presents a synthesis using the DsKabC [SEQ ID NO. 1] enzyme in lysate. Figure 3D presents a graph showing the conversion of prekainic acid to kainic acid over time in the synthesis of Figure 3C. Without being bound by theory, promiscuous kainoid synthases could be utilized late stage to cyclize novel kainoids from more synthetically feasible linear precursors. A chemoenzymatic synthesis of two novel kainoids wherein functional alkyne handles are installed onto isoprenoid precursors, condensed with L-glutamate, then cyclized via kainoid synthase enzymes is presented herein (Figure 4). This includes a fully synthetic method presented herein that forms an acyclic alkynyl-functionalized prekainic acid. This acyclic alkynyl-functionalized prekainic acid can be cyclized, for example, with the RadCl protein, such as the RadCl protein from Chondria armata, to form an alkynylfunctionalized prekainic acid (having a kainoid ring, a pyrrolidine ring). Overall, chemoenzymatic syntheses of neuroexcitatory and CuAAC-compatible kainoid analogs (clickable kainoid syntheses) are presented. These novel kainoids retain partial agonism of recombinant GluK2 channels, and can be further diversified and functionalized via copper catalyzed azide- alkyne cycloaddition (CuAAC) chemistry, enabling both rapid exploration of kainoid chemical space and for bio-orthogonal control of kainoid pharmacology in-vitro and in-vivo.9,10That is, functionalizing kainic acid with an alkyne, such as a terminal alkyne, allows for the alkyne-functionalized kainic acid to be reacted with an azide-functionalized group through the copper (Cu) catalyzed azide-alkyne cycloaddition (CuAAC) ("click") reaction. For example, an azide-functionalized fluorescent chemical group can be reacted with an alkyne-functionalized kainic acid to attach (covalently) the fluorescent chemical group through the triazole formed by the CuAAC reaction to the kainic acid. Labeling kainic acid can enhance the utility of kainic acid in studying neural phenomena. For example, labeling kainic acid with a fluorescent chemical group can allow identification of where in or on a cell (such as a kainate receptor), in or on an organ, and in an organism kainic acid binds. For example, functionalizing kainic acid with biotin can allow for binding to avidin or streptavidin of the functionalized kainic acid and the receptor or other protein to which the kainic acid is bound. That is, a synthesis of a derivative of the neuroexcitatory kainoid compound class functionalized with a terminal alkyne handle that enables bioorthogonal click chemistry via CuAAC with the potent kainoid pharmacophore is presented. A final step in this synthesis utilizes recombinant RadCl kainoid synthase enzyme from Chondria armata marine red macroalgae to catalyze (enzymatically transform) the bioactive pyrrolidine ring formation. The resulting alkyne- containing kainoid can then react with azides and copper catalysts to form kainoid analogs and install new functional handles. This establishes a rapidly tunable and scalable way to install fluorophores, photoaffinity labels, and other bioorthogonal handles to a potent kainate receptor agonist scaffold, which can serve as a pharmacological tools to study kainate receptors. The products made have bioorthogonal or “clickable” handles to further diversify the products following enzymatic construction. The methods presented avoid the use of protecting groups and comparatively harsh reagents. The enzymatic synthesis can be rapidly modified using processes like directed evolution to accept additional substrates and generate additional products. Utilizing the RadC 1 enzyme for preparative generation of kainoids avoids long chemical syntheses and is tolerant of non-native alkyne-bearing derivatives to generate novel kainoids. This transformation avoids the use of harsh and environmentally damaging reagents and is scalable. The methods do not require protein purification and can be performed in clarified lysates and whole cells containing the recombinant RadCl enzyme. Whereas photochromic and photo-affinity labeling compounds have been developed for other iGluRs, synthetic and biosynthetic methodologies are presented herein to endow neurobiologists with diversifiable tools designed specifically for this specific subtype to enable additional discoveries in KAR-mediated health and disease.11,12Definitions Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the provided compositions, suitable methods and materials are described below. Each publication, patent application, patent, and other reference mentioned herein is herein incorporated by reference in its entirety as if each such publication, patent application, patent, and other reference had been individually incorporated. In case of an inconsistency, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be limiting. The terms "identical" or percent "identity," in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (for example, about 80%, 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region, when compared and aligned for maximum correspondence over a comparison window or designated region). This definition also includes sequences that have deletions and / or additions, as well as those that have substitutions. For example, identity can be assesseds over a region that is at least about 25, 50, or 100 amino acids or nucleotides in length. Other features and advantages of the methods and compositions discussed herein will be apparent from the following written description, drawings, and claims. It is understood that wherever embodiments are described herein with the language "comprising", then otherwise analogous embodiments, described in terms of "containing", "consisting of”, and / or "consisting essentially of' are also provided. However, when used in the claims as transitional phrases, each should be interpreted separately and in the appropriate legal and factual context (e.g., in claims, the transitional phrase "comprising" is considered more of an open-ended phrase while "consisting of' is more exclusive and "consisting essentially of' achieves a middle ground). As used herein, the terms "approximately" and "about," as applied to one or more values of interest, refer to a value that is similar to a stated reference value. In certain embodiments, the term "approximately" or "about" refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11 %, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1 %, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value). For example, when used in the context of an amount of a given compound in a composition, "about" may mean + / -10% of the recited value. For instance, a composition including a compound having about 40% of a given compound may include 30-50% of the compound. The term "and / or" as used in a phrase such as "A and / or B" herein is intended to include each of the following: both A and B; A or B; A (alone); and B (alone). Likewise, the term "and / or" as used in a phrase such as "A, B, and / or C" is intended to encompass each of the following: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone). Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. Where embodiments of the disclosure are described in terms of a Markush group or another grouping of alternatives, the disclosed composition or method encompasses not only the entire group listed as a whole but also encompasses each member of the group individually and all possible subgroups of the main group and also encompasses the main group absent one or more of the group members. The disclosed methods and compositions also envisage the explicit exclusion of one or more of any of the group members in the disclosed compositions or methods. Unless otherwise indicated or indicated by the context, a singular form is to be understood as also encompassing the plural form, and a plural form is to be understood as also encompassing the singular form. An embodiment may include one or more described elements and may optionally include one or more additional elements that are not specifically described. An embodiment may be essentially free of or completely free of non-described elements; that is, non-described elements may optionally be essentially omitted or completely omitted from an embodiment. Embodiments Existing kainoid structure activity relationships denote the C4 side chain as the most tolerant to modification if the D system at C1’ is maintained (Figure 1).13Although substrate preferences of the kainoid synthases have yet to be thoroughly elucidated, successful biocatalytic and chemoenzymatic transformations with DsKabC [SEQ ID NO.1] and DabC (^-ketoglutarate- dependent dioxygenase) kainoid synthases are likewise tolerant to modest modifications of the isoprenoid chain that then becomes the kainoid C4 side chain post-cyclization.14–16We hypothesized that installation of a small hydrophobic acetylene handle on the isoprenoid chain distal from the glutamic acid moiety had the best chance of retaining both substrate activity with the kainoid synthases and agonistic activity with kainite receptors once cyclized. To this end, the terminal alkene of geranyl acetate (n = 1) was epoxidized using m-chloroperoxybenzoic acid (mCPBA) and oxidatively cleaved with sodium periodate (NaIO4). The resulting aldehyde underwent homologation with dimethyl 1-diazo-2-oxopropylphosphonate with concomitant acetate ester methanolysis to construct a terminal alkyne-functionalized prenol (n = 1, S3).17Dess Martin oxidation (for example, with Dess-Martin periodinane (DMP) in a chlorinated solvent such as dichloromethane (DCM)) resulted in an aldehyde (n = 1, S4). Subsequent reductive amination withL-glutamatic acid generated the desired propyne-extended derivative as a mixture of cis / trans diastereomers, referred to here on as “pre-alkynic acid” (n = 1, 2) a portmanteau of “alkyne” and “pre-kainic acid” (Figure 5A). The same synthetic workflow was applied to farnesyl acetate18to generate (with the following intermediate compounds: n = 2, S5; n = 2, S6; n =2, S7; n = 2, S8) an analogous prenyl-extended alkyne variant, named “pre-aldynic acid” (3) given its similar chain length to the dainic acid family of kainoid natural products (Figure 5A). Additionally, a trans- selective N-alkylation route for making other substrates in mechanistic studies of kainoid synthases was adapted19(Figure 5B). Although selective production of the trans pre-kainoid substrates was successful, poor conversion, competing dialkylation, and lactamization during ester deprotection decreased the utility of this alternate synthetic route. To assess whether wild type kainoid synthases were capable of cyclizing non-native alkyne-containing substrates, six kainoid synthase (enzyme) orthologs (DsKabC [SEQ ID NO.1], GfKabC [SEQ ID NO. 2], ReKabC [SEQ ID NO. 3], PpKabC [SEQ ID NO. 4], PmDabC [SEQ ID NO.5], CaRadC1 [SEQ ID NO.6]) were expressed in BL21 (DE3) E. coli (that is, from these E. coli into the medium) and prepared as clarified lysates.20Organisms other than or in addition to E.coli could be used to express a kanoid synthase, such as another bacterium, yeast, algae, or seaweed cells. Following the individual incubation of 2, 3, and native substrates with 75% v / v lysates supplemented with all necessary cofactors and co-substrates, assays were evaluated by ultra-performance liquid chromatography mass spectrometry (UPLC-MS) for putative cyclic / dehydrogenated and hydroxylated products (Figures 6A-6C). Canonical activity of all six kainoid synthase lysates with their respective native substrates was recapitulated: all four KabC enzymes (i.e., DsKabC [SEQ ID NO. 1], GfKabC [SEQ ID NO. 2], ReKabC [SEQ ID NO. 3], PpKabC [SEQ ID NO.4]) performed various degrees of cyclization with pre-kainic acid (R = H) (4), while PmDabC [SEQ ID NO.5] and CaRadC1 [SEQ ID NO.6] exhibited preferential activity for the geranyl-based substrates L-NGG (5) and 7’-carboxy-L-NGG (6). Lysate expressing the RadC1 gene from red macroalgae Chondria armata (CaRadC1 [SEQ ID NO.6]) exhibited modest reactivity with both pre-alkynic acid 2 and pre-aldynic acid 3; exhibiting relaxed substrate specificity compared to its homologues. In vitro reaction conditions were optimized with purified RadC1 to fully consume the cis / trans isomeric substrate mix of 2 or 3 (Figures 6D-6O), then scaled up to the multi-milligram level (20 mg) to isolate products by reversed-phase high- performance liquid chromatography (RP-HPLC). Major and minor cyclized products in addition to hydroxylated products were collected and characterized by 1D and 2D nuclear magnetic resonance (NMR) and high-resolution mass spectrometry (HRMS), validating that the major isolated products following the incubation of 2 and 3 with RadC1 were cyclic kainoids that retained the native pyrrolidine ring stereochemistries (Figures 7A-7G). Surprisingly, the major isolated cyclized products diverged at the newly formed C1’ alkene stereochemistry as determined by nuclear Overhauser effect spectroscopy (NOEs): the product isolated from RadC1 incubation of 2 contained a trans double bond reminiscent of the dainic and isodomoic acid B series; while the product isolated from incubation with 3 exhibited cis geometry matching that of the isodomoic and dainic acid A series (Figure 8). Based on established nomenclature rules, we proposed to name these major products alkynic acid B (7) and aldynic acid A (8) respectively.21Consistent with previous literature reports, β-hydroxylated products were also isolated as a major by-product (Figures 7E and7G).22Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) was directly performed with alkynic acid B (7) and aldynic acid A (8) following analytical enzymatic reaction by incubation with copper sulfate, additional ascorbate, and various azides directly into the quenched enzymatic assay, highlighting the rapid diversification potential of combining kainoid synthase enzymology with biorthogonal chemistry (Figures 8A-8D). To assess whether 7 and 8 maintained KAR activity, whole-cell patch clamp electrophysiology was employed on HEKT-293 cells transiently expressing rat GluK2. To compare the relative potency of these new analogs, kainic acid (1), dainic acid A (9), and isodomoic acid A (10), were likewise enzymatically synthesized as previously described.20Specifically, outside-out patches expressing GluK2 were excised from transfected HEK cells and GluK2 receptors were rapidly activated by piezo-driven fast jumps from a standard saline solution into 10 mM L-glutamate for 500 ms as a control. (PMID: 21307250). In the same patch, one or more kainoid ligands were applied at 1 mM (Figure 9A) and the relative peak response, normalized to full agonist glutamate (Figure 4B), current rise-time, desensitization decay and steady-state or equilibrium current were measured for each ligand (Figure 4C-E). Consistent with past work (PMID: 21307250), 1 was a partial agonist with about 40% peak response to glutamate (42 ± 3% glutamate peak, n = 10 patches). 9 had yet to be pharmacologically characterized and showed slightly lower efficacy (33 ± 4%, n = 10) compared to 1. Isodomic acid A, similar to domoic acid (PMID: 21307250), was a weak partial agonist showing 23 ± 4% (n = 5). To our excitement, both 7 and 8 showed partial agonism was retained but with reduced efficacy compared to native analogues (11 ± 2%, 7± 1%, n = 6). We are further encouraged to observe that C1’ double bond stereochemistry appears to have a weak affect, if any, on the activity of these novel derivatives in contrast to established structure activity relationships observed with native kainoids.5Glutamate, 1 and 9 all showed comparable rise-times and desensitization decays in the 3-6 ms range, and near-complete desensitization (Figures 9C-9E). Both non-native kainoids alkynic acid B 7 and aldynic acid A 8 showed slower rise times, desensitization decays, and less complete desensitization compared to their natural analogues (Figures 9C-9E). 10 showed the slowest rising phase and desensitization decays, as well as reduced desensitization (Figures 9C- 9E). Docking of alkynic acid B 7 and aldynic acid A 8 suggest that retention of the kainoid ring pharmacophore allows these analogues to retain key interactions with the GluK LBD (Figures 10D and 10E). Retention of the kainoid ring stereochemistry putatively maintains key electrostatic interactions with R523, E738, and N721, while unsaturation at the C1’ position preserves D-D stacking with Y488. Importantly, docking suggests that the terminal alkyne handles likely stick out of the LBD with aldynic acid’s extended chain being particularly solvent-accessible and theoretically available for affinity labeling (Figures 10D and 10E). Docking of kanoids is also shown in Figures 10A – 10C, 10F, and 10G. A probe can be attached to a kanoid with the click chemistry (CuAAC) presented herein. For example, such a probe can be a dye, a binding agent, or another label. For example, such a probe can include fluorescein (FI), Cy5, Alexa 488 (Ax488), Alexa 647 (Ax647), biotin (Bt), or SeTau-647 (ST647). A chemoenzymatic synthesis of novel KAR agonists with the ability for rapid diversification and construction of new biorthogonal tools is described. CaRadC1’s tolerance for extended substrates enables its use for engineering and creation of additional novel kainoids with improved pharmacological properties. Competing cyclization and hydroxylation reaction pathways dependent on substrate double bond stereochemistry contributes to understanding kainoid synthase enzymology. Enzymatic construction of the kainoid ring pharmacophore bypasses costly steps and harsh reagents, enabling late-stage diversification to efficiently expand kainoid chemical space. CuAAC compatibility enables expansion of kainoid diversity and contributes to the development of KAR subtype-specific agents.25Materials, Biochemical Methods, and Analytical Methods All chemicals, solvents, and media components were used as received from commercial suppliers (Millipore Sigma, Thermo Fisher Scientific, Enamine). Reagents were used without further purification unless noted otherwise. Synthetic and analytical methods Anhydrous reactions were done in oven-dried glassware and carried out under inert argon atmosphere. Reactions were monitored used thin-layer chromatography (TLC, Merck, 60 F254) and visualized under a 254 nm UV lamp before staining with either potassium permanganate (1.5 g KMnO4, 10 g K2CO3, 1.25 mL 10% NaOH in 100 mL DI water), 2,4-DNP (12 g 2,4- dinitrophenylhydrazine, 60 mL H2SO4, 80 mL DI water, 200 mL 95% EtOH), PMA (10 g PMA in 100 mL 95% EtOH), or ninhydrin staining solutions (1.5 g ninhydrin, 3 mL AcOH, 100 mL n- butanol). TLC Rf values were rounded to the nearest 0.05. Silica (Alfa Aesar, 60 (215-400 mesh)) was used as the stationary phase when flash column chromatography was employed to purify compounds. Rotary evaporation was used to concentrate samples under reduced pressure and subsequent lyophilization was used to dry polar products from aqueous solutions and to remove trace impurities. Nuclear magnetic resonance (NMR) spectra were obtained using an Avance III HD spectrometer (Bruker) equipped with a BBFO SmartProbe at 500 MHz (1H NMR) or 125 MHz (13C NMR) using CDCl3, CD3OD, or D2O as solvents. Chemical shifts (E) are reported in ppm and referenced to the residual CDCl3solvent signal (δ = 7.26 ppm for 1H, δ = 77.2 ppm for13C NMR) for samples in CDCl3or referenced to the residual CD3OD signal (δ = 3.31 ppm for 1H, δ = 49.0 ppm for 13C NMR) for samples in CD3OD. Samples in D2O are referenced either to MeOH (δ = 3.31 ppm for 1H, δ = 49.0 ppm for 13C NMR) or formic acid (δ = 8.26 ppm for 1H, δ = 166.3 ppm for 13C NMR) as an internal standard. NMR data are reported as follows: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, J = coupling constant in Hz. LCMS, HPLC, and FPLC instrumentation General LCMS measurements were measured on a Bruker Elute UHPLC system coupled with a Bruker amazon SL ESI-Ion Trap mass spectrometer in negative mode. Compounds were separated via reversed-phase chromatography on a Bruker Intensity Solo C18(2), 2 μm- 2 x 100 mm column with the eluents water + 0.1% formic acid (Solvent A) and acetonitrile + 0.1% formic acid (Solvent B). LC methods used a flow rate of 0.3 mL / min and a general gradient: 10% to 20% B over 3 minutes, 20% to 45% B over 3 min, 45% - 100% B over 2 min, hold at 100% B for 2 min, 100% - 10% B over 1 min, hold at 10% B for 2 min. Preparative HPLC purification was performed using a Shimadzu Prominence preparative HPLC system with a SPD-20A model UV / Vis detector monitored at both 210 nm and 254 nm. Semipreparative and analytical HPLC purification was performed using an Agilent 1260 Infinity system with a G1314B model VWD. The monitored wavelength was 210 nm for all runs. High- performance flash chromatography was performed using a Biotage Isolera Prime system with a dual variable UV detector (200 - 400 nm) monitored at 210 nm and 254 nm. Purification of recombinant 6xHis tagged proteins was performed on an AKTAGo instrument (GE Healthcare) operated in a 4 ºC fridge equipped with buffers filtered through a 0.22 μm nitrocellulose membrane and degassed under vacuum. FPLC data was analyzed with UNICORN v 7.5 (GE Healthcare). Expression of kainoid synthases in BL21 (DE3) E. coli for clarified lysate screening Glycerol stocks of kainoid synthase transformed BL21(DE3) cells were used to inoculate 25 mL of LB media supplemented with 50 μM of kanamycin. The cultures were grown at 37 ºC and 200 RPM overnight, then subsequently diluted 1:75 into 1 L cultures of Terrific Broth supplemented with 50 μM Kanamycin for large scale expression. Each 1 L culture was incubated at 37 ºC and 200 RPM until a 1 mL representative sample OD600reached ~0.6 – 0.8. Once the desired density was reached, the cultures were cooled to 18 oC for 30 – 60 minutes before adding IPTG to induce expression. CaRadC1 [SEQ ID NO. 6] cultures were induced to a final IPTG concentration of 1 mM, all other cultures were induced with a final IPTG concentration of 0.5 mM. The cells were grown overnight at 18 ºC and 200 RPM before harvesting the cell mass via centrifugation at ~3500 x g for 20 minutes at 4 ºC. The supernatant was bleached and discarded. The pelleted cells were directly resuspended in 30 mL of ice-cold modified Gel Filtration buffer (“mGF” buffer, 100 mM HEPES pH 8.0, 100 mM KCl) for same day testing with substrates. One round of freeze thaw was performed by rapidly freezing in a methanol-dry ice bath for approximately 5 minutes, then thawed by incubation in room temperature deionized water for approximately 10 minutes before placing on ice. If cells were not being used same day, the cells were instead resuspended in storage buffer (50 mM HEPES pH 8.0, 100 mM NaCl, 30 mM imidazole, 10% glycerol) and stored at -70 ºC until needed. When using previously stored cells, they were likewise thawed in room temperature deionized water for approximately 30 minutes, then centrifugated at ~3500 x g for 20 minutes at 4 ºC to pellet the cell mass. The storage buffer was bleached and discarded before resuspending the cells in mGF buffer; these cells were then sonicated immediately without further freeze-thaw cycles. Prior to sonication, each cell suspension was diluted with mGF buffer to a corrected OD600of ~175 before lysing via sonication on an ice bath with magnetic stirring. Each cell line was lysed at 50% amplitude, alternating between 15 sec on and 45 seconds off (Q500 Sonicator (QSonica), 6.4 mm probe) until the corrected OD600had fallen below 100. CaRadC1 [SEQ ID NO. 6] was lysed for 30 – 40 minutes total (5 – 10 minutes duty time), while all other cell lines reached the desired OD or lower after only 10 minutes of total sonication time (2.5 minutes duty time). Each lysate was then clarified via centrifugation at ~16,000 x g and 4 ºC for 60 minutes. The clarified supernatant was then decanted into a fresh tube and kept on ice until needed for substrate screening assays. CaRadC1 FPLC affinity purification Expression of CaRadC1 [SEQ ID NO.6] transformed BL21 (DE3) E. coli cells was done as described above with minor modifications. Growth and expression were carried out identically at a 2 L scale, however after harvesting cells they were instead resuspended in 40 mL of lysis buffer (50 mM HEPES pH 8.0, 1 M NaCl, 30 mM imidazole, 3 mM EDTA, 10% glycerol). One freeze-thaw cycle was performed before diluting the cells to a corrected OD600of 150 before adding egg white lysozyme to a final concentration of 0.2 mg / mL. The suspension was then sonicated with stirring and on ice at 50% amplitude alternating between 15 sec on and 45 sec off for 20 minutes total (5 minutes duty time). Addition of lysozyme aided with inconsistent sonication results, giving a corrected OD600of 50 in half the usual time and increasing final soluble yields to 106 mg / L post affinity purification. The crude lysate was then clarified via centrifugation at ~16,000 x g and 4 ºC for 60 minutes. The clarified supernatant was then loaded onto a 5 mL HisTrap FF column (GE Healthcare Life Sciences) that was pre-equilibrated with at least 5 column column (CV) of buffer A (20 mM Tris-HCl pH 8.0, 1M NaCl, 30 mM imidazole). After loading, the column was washed with buffer A until the UV absorbance plateaued at or below 50 mAU. The column was then washed with 10% buffer B (20 mM Tris-HCl pH 8.0, 1 M NaCl, 250 mM imidazole) for 5 -10 CV to remove any non-specifically bound proteins. This 10% buffer B wash was collected and processed analogously as a significant amount of active and sufficiently pure CaRadC1 [SEQ ID NO.6] would be collected in this wash step and utilized to supplement purified enzyme stocks. CaRadC1 [SEQ ID NO.6] was then eluted from the HisTrap column using a linear gradient of 10% to 100% buffer B over 60 mL (12 CV’s).5 mL fractions were taken during elution with a flowrate of 2 mL / min throughout the purification process. Fractions were assessed for purity using SDS-PAGE (10% acrylamide) and fractions containing CaRadC1 [SEQ ID NO. 6] had a final 2 mM EDTA added to strip any iron or other metals that copurified with the enzyme. Fractions determined to be G 90% pure were combined and concentrated using an Amicon ultra centrifugal filter (30 kDa molecular weight cut off) before buffer exchanging through a PD-10 desalting column (GE Healthcare Life Sciences) pre-equilibrated with ice-cold mGF buffer via gravity filtration on ice. Enzyme concentration and yield was determined via Bradford assay and pure enzyme was either stored on ice and used immediately or aliquoted and stored at -70 ºC. Analytical assays of kainoid synthase-expressing clarified lysates Analytical lysate assays were conducted in a final concentration of 75% v / v of clarified lysate, 100 mM HEPES pH 8.0, 100 mM KCl, 10 mM B-ketoglutarate, 2 mM L-ascorbate, 0.5 mM FeSO4heptahydrate, and 1 mM of substrate cis / trans mix. The total reaction volume was 100 μL and lysate followed by FeSO4were the last additions each assay. Reactions were allowed to run for 16 h at room temperature on a tilting rack before quenching via the addition of 100 μL of 0.1 mM chloramphenicol in methanol (1 eq.) and centrifugation at ~ 16,000 x g for 10 minutes. The supernatant was then directly subjected to analysis by UPLC-MS. The LC conditions employed were optimized for each specific substrate tested (Table 1). In vitro assays of 2 and 3 with purified DsKabC, PmDabC, and CaRadC1 Analytical enzyme assays of pre-alkynic acid (2) and pre-aldynic acid (3) were conducted at a total volume of 100 μL with the optimized conditions determined in Figures 6F-6H and S6M- 6O. Final concentrations of each component were as follows: 100 mM HEPES pH 8.0, 100 mM KCl, 10 mM B-ketoglutarate, 2 mM L-ascorbate, 0.25 mM FeSO4heptahydrate, 1 mM of substrate cis / trans mix, and 0.1 mM of purified DsKabC [SEQ ID NO. 1], PmDabC [SEQ ID NO. 5], or CaRadC1 [SEQ ID NO. 6]. Each assay was run in triplicate with a single “no enzyme” negative control. Enzyme followed by FeSO4were the final additions to each assay before incubating at room temperature for 16 h on a tilting rack. Each reaction was then quenched with 0.1 mM chloramphenicol internal standard in methanol (1 eq.) and centrifuged at ~ 16,000 x g for 10 minutes. The supernatant was then directly subjected to UPLC-MS analysis. Enzymatic synthesis of alkynic acid B (7) and aldynic acid A (8) Preparative assays were conducted in the same manner as optimized analytical scale assays with some modification.13 separate reactions each in a 2 mL microcentrifuge tube was scaled to 1.2 mL (15.6 mL total reaction volume). The final concentration of each component was as follows and was added to each reaction in the following order: 100 mM HEPES pH 8.0, 100 mM KCl, 5 mM substrate cis / trans mix, 50 mM B-ketoglutarate (10 eq.), 10 mM L-ascorbate (2 eq.), 0.25 mM CaRadC1 [SEQ ID NO. 6] (0.05 equiv), and 1.25 mM FeSO4 heptahydrate. Each tube was capped lightly to allow for oxygen flow and infusion. After incubation on a tilting rack at rt for 16 h, the 13 reactions were combined and quenched by addition of an equal volume of methanol. The solution was vortexed and centrifugated at ~10,000 x g for 20 minutes to precipitate protein. The supernatant was then filtered through a 0.22 μm cellulose acetate filter, concentrated in vacuo and subjected to RP-HPLC purification via HPLC procedure A for alkynic acid B (7) or HPLC procedure B for aldynic acid A (8). Alkynic acid B (7) was obtained as a fluffy white powder.1H NMR (500 MHz, Deuterium Oxide) E 5.21 (t, J = 6.7 Hz, 1H), 4.00 – 3.95 (m, 1H), 3.53 (dd, J = 12.0, 7.5 Hz, 1H), 3.37 (t, J = 11.5 Hz, 1H), 3.06 – 2.99 (m, 1H), 2.99 – 2.93 (m, 1H), 2.90 (d, J = 4.7 Hz, 2H), 2.34 (d, J = 7.2 Hz, 2H), 2.31 – 2.27 (m, 1H), 1.61 (s, 3H). LRMS (ESI) calculated for C13H17NO4251.12, found 250.02 [M-H]-. Aldynic acid A (8) was obtained as a sticky white powder.1H NMR (500 MHz, Methanol-d4) E 5.39 (t, J = 7.1 Hz, 1H), 5.23 – 5.11 (m, 1H), 4.01 (d, J = 4.8 Hz, 1H), 3.58 – 3.48 (m, 2H), 3.44 – 3.36 (m, 1H), 3.02 (dq, J = 11.8, 6.7 Hz, 2H), 2.80 (dt, J = 15.5, 7.6 Hz, 1H), 2.69 (t, J = 7.0 Hz, 1H), 2.66 (s, 0H), 2.40 (dq, 2H), 2.29 – 2.21 (m, 3H), 2.21 – 2.14 (m, 4H), 1.72 (s, 3H), 1.64 (s, 3H). LRMS (ESI) calculated for C18H25NO4319.18, found 318.06 [M-H]-. Enzymatic synthesis of kainic acid (1) Kainic acid (1) was synthesized from pre-kainic acid as reported with modifications.1Clarified DsKabC [SEQ ID NO.1] lysate was prepared as described above. A 1 L round bottom flask was charged with 1.331 g of B-ketoglutaric acid disodium salt (7.00 mmol, 100 mM), 0.123 g of L-ascorbic acid (0.70 mmol, 10 mM), and 0.150 g of prekainic acid (0.70 mmol, 10 mM).45 mL of aqueous buffer was added (100 mM HEPES pH 8.0, 300 mM KCl) and the solution was stirred until all solids were dissolved. 25 mL of clarified DsKabC [SEQ ID NO. 1] lysate was poured into the reaction (36 % v / v) before adding 0.039 g of FeSO4heptahydrate (0.14 mmol, 2 mM). The reaction was lightly stoppered to allow for oxygenation and was stirred at 23 ºC until no substrate was detected by UHPLC-MS. After 20 hrs, an equal volume of MeCN (70 mL) was added to quench the reaction. The solution was then centrifuged at ~ 4,000 x g for 15 minutes at 23 ºC to precipitate the spent protein and other insoluble material. The supernatant was collected, and the pellet resuspended in 15 mL of deionized H2O and subjugated to a second round of centrifugation. The supernatants were pooled and concentrated in vacuo to obtain 12.263 g of sticky brown wax.3 gs of this crude extract was adsorbed into 5 g of celite, and dry loaded onto a 25 g SFAR Silica D 60 Biotage column that was pre-equilibrated with 10 CVs of 5 % H2O in MeCN + 0.1 % FA. Initial purification was done according to Flash procedure C. Fractions containing kainic acid were identified via UHPL-MS, pooled, and concentrated in vacuo yielding 0.555 g of light-yellow wax. This material was then further purified by HPLC procedure A to yield 0.005 g of kainic acid (1) as a white fluffy powder.1H NMR characterization matched previous reports: (500 MHz, Deuterium Oxide) E 5.01 (s, 1H), 4.06 (d, J = 2.8 Hz, 1H), 3.60 (dd, J = 11.7, 7.4 Hz, 1H), 3.40 (t, J = 11.4 Hz, 1H), 3.08 – 3.02 (m, 1H), 3.02 – 2.94 (m, 1H), 2.45 – 2.28 (m, 2H), 1.73 (s, 3H); LRMS (ESI) Calculated for C10H15NO4213.10, found _ (M+_)_.26Enzymatic synthesis of dainic acid A (9) Dainic acid A (9) was synthesized from N-geranyl-L-glutamic acid as reported with modifications.15Clarified CaRadC1 [SEQ ID NO.6] lysate was prepared as described above then dialyzed for 2 hrs at 23 ºC against 0.1 mM HEPES pH 8.0 and 0.3 mM KCl in 10 kDa MWCO tubing to remove low molecular weight contaminants. A 1 L round bottom flask was charged with 2.851 g of B-ketoglutaric acid disodium salt (15.00 mmol, 100 mM), 0.264 g of L-ascorbic acid (1.50 mmol, 10 mM), and 0.204 g of N-geranyl-L-glutamic acid (0.70 mmol, 5 mM).113 mL of aqueous buffer was added (100 mM HEPES pH 8.0, 300 mM KCl) and the solution was stirred until all solids were dissolved.37 mL of dialyzed CaRadC1 [SEQ ID NO.6] lysate was poured into the reaction (25 % v / v) before adding 0.042 g of FeSO4heptahydrate (0.15 mmol, 1 mM). The reaction was lightly stoppered to allow for oxygenation and was stirred at 23 ºC until the reaction was deemed complete by UHPLC-MS analysis. After 20 hrs, an equal volume of MeCN (150 mL) was added to quench the reaction. The solution was then centrifuged at ~ 4,000 x g for 15 minutes at 23 ºC to precipitate the spent protein and other insoluble material. The supernatant was collected, and the pellet resuspended in 15 mL of deionized H2O and subjugated to a second round of centrifugation. The supernatants were pooled and concentrated in vacuo to obtain 7.295 g of sticky brown wax. 4 gs of this crude extract was purified according to Flash procedure D. Fractions containing 9 were identified via UHPL-MS, pooled, and concentrated in vacuo yielding 0.041 g of a dainic acid A as a white fluffy powder.1H NMR characterization matched previous reports2(500 MHz, Methanol-d4) E 5.38 (t, J = 7.1 Hz, 1H), 5.05 (t, J = 6.9 Hz, 1H), 3.94 (d, J = 6.3 Hz, 1H), 3.57 (dt, J = 15.4, 8.0 Hz, 2H), 3.37 (td, J = 12.4, 5.3 Hz, 1H), 3.01 (p, J = 6.9 Hz, 1H), 2.75 (dt, J = 15.4, 7.5 Hz, 1H), 2.68 – 2.55 (m, 2H), 2.41 (dd, J = 16.3, 7.8 Hz, 1H), 1.72 (s, 3H), 1.67 (s, 3H), 1.62 (s, 3H); HRMS (ESI) Calculated for C15H23NO4281.1627, found 280.1554 [M-H]-. Enzymatic synthesis of isodomoic acid A (10) Isodomoic acid A (10) was synthesized from 7’-COOH-N-gernayl-L-glutamic acid as described previously with little modification.23Purified PmDabC [SEQ ID NO. 5] was prepared as described above. The reaction was split equally into 3 parallel reactions each with a final volume of 5 mL prepared in 15 mL falcon tubes. The final concentration of each component was as follows and was added to each reaction in the following order: 100 mM HEPES pH 8.0, 100 mM KCl, 5 mM 7’-COOH-N-gernayl-L-glutamic acid cis / trans mix, 10 mM B-ketoglutarate (2 eq.), 2 mM L- ascorbate (0.4 eq.), 0.07 mM PmDabC [SEQ ID NO. 5] (0.015 equiv), and 0.05 mM FeSO4heptahydrate (0.01 equiv). Each tube was capped lightly to allow for oxygen flow and infusion. After incubation on a tilting rack at 23 ºC for 16 h, the 3 reactions were combined and quenched by addition of an equal volume of methanol. The solution was vortexed and centrifugated at ~10,000 x g for 20 minutes to precipitate protein. The supernatant was then filtered through a 0.22 μm cellulose acetate filter, concentrated in vacuo, and subjected to RP-HPLC purification via HPLC procedure B to afford 0.006 g of isodomoic acid A as a white fluffy powder.1H NMR characterization matched previous reports2(500 MHz, Deuterium Oxide) E 6.58 (t, J = 7.1 Hz, 1H), 5.47 (t, J = 7.3 Hz, 1H), 3.91 (d, J = 7.2 Hz, 1H), 3.67 – 3.61 (m, 1H), 3.57 (q, J = 7.5 Hz, 1H), 3.40 (dd, J = 11.4, 7.7 Hz, 1H), 2.99 – 2.88 (m, 2H), 2.84 – 2.76 (m, 1H), 2.57 (dd, J = 16.3, 6.2 Hz, 1H), 2.41 (dd, J = 16.3, 8.7 Hz, 1H), 1.77 (s, 3H), 1.70 (s, 3H). LRMS (ESI) Calculated for C15H21NO6311.14, found _ (M+_)_. Analytical CuAAC of 7 and 8 CuAAC was performed directly in quenched analytical assay media described above. 75 μL of the quenched enzymatic assay supernatants containing 2.5 mM of alkyne-functionalized substrates and cyclized / hydroxylated products were diluted to a final volume of 375 uL and final concentration of 1 mM. Benzyl azide, CuSO4pentahydrate, and sodium ascorbate were added to a final concentration of 1 mM, 0.2 mM, and 0.6 mM respectively. Additional controls lacking CuSO4or sodium ascorbate were prepared to assess if residual ascorbate from the enzymatic transformation could sufficiently drive CuAAC activity. Each reaction was incubated at rt for 4 h before analysis via UPLC-MS (Figures 8B-8D). Preparative CuAAC of alkynic acid B-benzyl triazole (11) and alkynic acid B-coumarin triazole (12) To a dram vial charged with a stir bar was added alkynic acid B (7) (0.003 g, 25 mM, 1.0 equiv), either benzyl azide or 7-azido-4-methylcoumarin (25 mM, 1.0 equiv), copper sulfate pentahydrate (5 mM, 0.20 equiv), and sodium ascorbate (15 mM, 0.60 equiv) in 0.320 mL of 33% aqueous t-butanol. The reaction was covered in aluminum foil and stirred at 23 oC for 4 hr before concentrating in vacuo. The resultant crude material was purified via UHPLC-MS procedure E. Less than 0.001 g of click material was obtained and 1H NMR of benzyl triazole product was obtained.1H NMR (800 MHz, Deuterium Oxide) E 7.70 (s, 1H), 7.35 (dt, J = 13.4, 7.0 Hz, 3H), 7.25 (d, J = 7.3 Hz, 2H), 5.50 (s, 2H), 5.27 (t, J = 7.5 Hz, 1H), 3.93 (d, J = 2.8 Hz, 1H), 3.70 (t, J = 6.0 Hz, 1H), 3.56 (dd, J = 11.6, 4.2 Hz, 2H), 3.47 (dd, J = 11.6, 6.7 Hz, 3H), 3.40 (d, J = 6.9 Hz, 2H), 3.33 (t, J = 11.3 Hz, 1H), 2.97 – 2.88 (m, 2H), 2.06 – 1.96 (m, 2H), 1.64 (s, 3H); LRMS (ESI) Calculated for C20H24N4O4384.18, found 383.12 (M-H)-. LRMS calculated for alkynic acid B, coumarin triazole click product C23H24N4O6452.17, found 451.19 (M-H)-. GluK2 Pharmacological and Docking Methods Outside-out patch-clamp recordings of kainoids with GluK2 homomeric channels All kainoids tested in this study were 80 to >95% pure or greater by 1H NMR and > 70% pure by analytical HPLC prior to testing. HEK293T cells were maintained in Dulbecco’s modified Eagle’s medium with 4.5 g / liter glucose, L-glutamine, and sodium pyruvate (Corning / Mediatech, Inc.) or Eagle’s minimum essential medium with Glutamax & Earle’s Salts (Gibco), supplemented with 10% FBS (Atlas Biologicals) and penicillin / streptomycin (Invitrogen). Cells were passaged every 2–3 d when ~90% confluence was achieved. Cells were plated on tissue culture–treated 35 mm dishes, transfected 24 hours later with 1 μg GluK2 and 0.2 μg eGFP plasmids per dish using PEI (3:1 ratio). Excised patch recordings were obtained 24-48 hours post-transfection. Outside-out patches were excised using heat-polished, thick-walled borosilicate glass pipettes of 3 – 15 MH resistance. The pipette internal solution contained (in mM) 135 CsF, 33 CsOH, 11 EGTA, 10 HEPES, 2 MgCl2, and 1 CaCl2(pH 7.3 with CsOH). External solutions was 150 NaCl, 10 HEPES, 1 CaCl2, and 1 MgCl2, (pH 7.4 with NaOH). All recordings were performed at room temperature with a holding potential of −60 mV using an Axopatch 200B amplifier (Molecular Devices). Data were acquired using Clampex10.7, at 25 kHz with 10 kHz filtering. Series resistance was routinely compensated by 90 – 95% where the peak amplitude exceeded 100 pA. Rapid perfusion was performed using home-built, double- or triple-barrel application pipettes (Vitrocom). Application pipettes were translated using piezo actuators driven by voltage power supplies. The command voltages were generally low-pass–filtered (50–100 Hz, eight-pole Bessel). Molecular docking of kainoids into GluK2 LBD SMILES models of alkynate B and aldynate A were docked into the crystal structure of kainate-bound GluK2 LBD (PDB ID 2XXT)23using DynamicBind24on the neurosnap online portal. Models were compared to structures of glutamate (2XXR), kainate (2XXT), and domoate (8GC2) bound to GluK2 LBDs and visualized in PyMOL. For all RP-HPLC procedures, the crude material was resuspended in a mixture H2O / MeCN buffered with 0.1% formic acid matching the initial column equilibration conditions. If necessary, additional formic acid was added to the adjust the pH between 4 – 5 by Whatman paper. Each sample was centrifugated at 16,000 x g for 10 minutes to remove insoluble material prior to injection.

[0048] Table 1. LC conditions for preparative and semi-preparative purifications

[0049] Chemical Syntheses (E)-5-(3,3-dimethyloxiran-2-yI)-3-niethylpent-2-en-l-yl acetate (SI): Compound SI was prepared according to literature17: To a solution of geranyl acetate (2.925 g, 14.90 mmol) in DCM (20 mL) at 0 °C was added a solution of m-CPBA (3.600 g [<77% purity], 15.65 mmol) in DCM (40 mL) dropwise via addition funnel. The mixture was stirred at 0 °C for an additional 3 h. The initially clear colorless solution rapidly turned an opaque milky white upon m-CPBA addition, which remained throughout the duration of the mixing time. After 3 h, TLC confirmed the consumption of starting material, and a saturated aqueous solution of NaHCO3(75 mL) w as added and the layers were separated. The aqueous phase was further extracted with DCM (2 x 60 mL), and pooled organic layers ivere washed with brine (75 mL), dried over MgSO4, filtered, and concentrated in vacuo. The crude reaction mixture was purified over a short plug of silica using a 4: 1 hexanes :EtO Ac eluant system. Pooled fractions were concentrated in vacuo yielding a clear colorless oil in 88% yield (2.799 g, 13.18 mmol). Rf= 0.70 (3: 1 hexanes :EtOAc):1H NMR (500 MHz, CDCl3) δ 5.41 - 5.35 (m, 1H). 4 58 (d, J = 7. 1 Hz, 2H), 2.68 (t, J = 6.2 Hz. 1H), 2.18 (ddq, J = 30.1, 14.6, 7.7 Hz, 2H), 2.03 (s. 3H), 1.71 (s, 3H), 1.68 - 1.61 (m, 2H). 1.29 (s, 3H), 1.25 (s, 3H):13C NMR (500 MHz, CDCl3) δ 171 1, 141.3, 119 0, 64 0. 61 3. 58 4, 36.2, 27.1, 24.9, 21.1,

[0050] 18.8, 16.5.

[0051] (E)-3-methyI-6-oxohex-2-en-l-yl acetate (S2): Compound S2 was prepared according to literature1': To a solution of epoxide S1 (2,511 g, 11 .828 mmol) in THF:H2O (10: 1 , 20 mL) at 0 °C was added NalOr (1.518 g, 7.097 mmol) followed by HIO4·2H2O (2.966 g, 13.011 mmol). The mixture was stirred in an ice-water bath for 1 hr before allowing to cool to room temperature over

[0052] 1 .5 hrs. The reaction mixture was quenched -with saturated aqueous NaHCO3(25 mL), poured into H2O (25 mL) and extracted with EtOAc (3 x 50 ml.,). The combined organic layers were washed with bnne (2 x 50 mL), dried over Na2SO4, filtered and concentrated in vacuo. The crude material was purified by silica flash chromatography (4: 1 hexanes: EtOAc) to give a light yellow oil in 79% yield (1.590 g, 9.341 mmol). Rf= 0.33 (4: 1 hexanes:EtOAc);1H NMR (500 MHz, CDCl3) δ 9.71 (d. J = 1.8 Hz, 1H), 5.30 (tt, J = 7.0, 1.4 Hz, 1H), 4.51 (d, J = 7.1 Hz, 2H), 2.52 (td, J = 7.6,

[0053] 1.6 Hz, 2H), 2.32 (t, J = 7.6 Hz, 2H), 1.98 (d, 7 = 1.0 Hz, 3H), 1.66 (s, 3H).

[0054] (E)-3-methyIhept-2-en-6-yn-l-ol (S3): Compound S3 was prepared as follows17: To a round bottom flask charged with a stir bar and K2CO3(0 600 g, 4.338 mmol) was added a solution of S2 (0.369 g, 2. 169 mmol) in HPLC grade MeOH (15 ml.,) and allowed to stir for 15 min. DAMP (0.500 g, 2.603 mmol) was added via syringe and stirred at room temperature for 18 hrs. The reaction mixture was diluted with Et2O (40 mL), washed with 5% aqueous NaHCO3(20 mL), brine (20 mL), dried over anhydrous NasSO4, filtered and concentrated to afford a light-yellow oil (0.217 g, 1 .747 mmol) in 81 % yield. Rf= 0.65 (3:2 hexanes: EtOAc): NMR (500 MHz, CDCl3) δ 5.44 (t, 1H), 4.14 (d, .7 = 6.8 Hz, 2H), 2.31 (dt, J = 7.1, 2.1 Hz, 2H), 2.29 (dd, J = 2.4, 1.3 Hz, 1H), 2.23 (t, J = 7 A Hz, 2H), 1.95 (t, J = 2.6 Hz, 1H), 1.67 (s, 3H).

[0055] (E)-3-methyIhept-2-en-6-ynal (S4): Compound S4 was prepared as follows22: Dess Martin penodinane (1.291 g, 3.044 mmol) was added to a solution of S3 (0.189 g.1.522 mmol) in dry CH2Cl2(7 mL) at once and stirred at room temperature for 1 h. The reaction mixture was quenched with an aqueous 10% sodium thiosulfate solution (7 mL) and washed with water (15 mL). Organic compounds were extracted from pooled aqueous layers with CH2Cl2(3 x 15 mL). Pooled organic layers were washed with brine (15 mL). dried over MgSO4, filtered and concentrated in vacuo The crude reaction mixture was purified by silica flash chromatography (3: 1 hexanes :EtO Ac), yielding S4 as a clear yellow oil (0.075 g, 0.614 mmol, 40%). Rf= 0.70 (2: 1 hexanes :EtO Ac);1H NMR (500 MHz, CDCl3) δ 10.01 (d, J = 7.9 Hz, 1H). 5.92 (d, J = 7.9, 1.3 Hz, 1H), 2.45 - 2 41 (m, 4H). 2. 19 (s, 3H). 2.01 - 1.98 (ra. 1H);13C NMR (500 MHz. CDCl3) 153.21 , 128.43. 39 46, 26.56, 18.21 , 17.09.

[0056] (3-methylhept-2-en-6-yn-l-yl)-L-glutaniic acid (2): A procedure was adapted from the literature22as follows: A solution of S4 (0.062 g, 0 508 mmol) in methanol (2.5 mL) was added dropwise to an aqueous solution (2.5 mL) of L-glutamic acid (0.090 g, 0.609 mmol) and sodium hydroxide (0.049 g, 1.22 mmol) and stirred for 3.5 hrs at room temperature. The reaction mixture was cooled to 0 °C, sodium borohydride (0.029 g, 0.762 mmol) was added at once, and the reaction mixture was stirred for an additional Ih. Formic acid was added until pH 4, and the reaction mixture was washed with toluene before concentration in vacuo. The crude material was then purified following Flash Procedure A. Fractions containing desired products were identified by UHPLC-MS, pooled, concentrated in vacuo and lyophilized, affording a white solid (0.060 g, 0.237 mmol, 47%, both regioisomers).1H NMR (500 MHz, D2O) δ 5.32 (dt, J = 25.6, 7.7 Hz, 1H), 3.67 (dq, J = 12.8. 6.4. 5.0 Hz. 2H), 3.59 (dd, J = 11.9. 6.0 Hz, 1H). 2.41 (q, J = 7.2 Hz, 2H). 2.37 - 2.27 (m, 4H), 2.24 (t, J = 6.9 Hz, 1H), 2.04 (ddp, J = 21.8, 14.4, 6 8 Hz, 2H), 1.75 (s, 1H), 1.66 (s, 2H);13C NMR (500 MHz, D2O) 178.35, 173.18, 145.51, 145.16, 115.38. 114.22, 85.20, 84.89, 69.92, 69.82, 60.63, 60.10, 43 97, 43.78, 37.25. 31.3.3, 29.87, 25.37, 25.34, 22.14, 16 21, 16.15, 15.46; HRMS (ESI) Calculated for C13H19NO4253.1314, found 252.1243 [M-H]-.

[0057] (E)-7-bromo-5-methylhept-5-en-l-yne (S9): Compound S9 was prepared according to literature27: To a solution of S3 (0.300 g, 2.42 mmol) in ether at 0 °C was added PBr3(0.327 g, 1.21 mmol) dropwise under argon atmosphere. After 1 h the reaction mixture was treated with aqueous NaHCO3(20 mL) and extracted with ether (3 x 20 mL). The combined organic extract was washed with brine, dried over Na2SO4, and concentrated in vacuo to give S9 (0.249 g, 1.33 mmol) in 55% yield.1H NMR (500 MHz, CDCl3) δ 5.62 (t, J - 8.6 Hz, 1H), 4.03 (d, J = 8.4 Hz, 2H), 2.42 - 2.27 (m, 4H), 1.98 (t, J = 2.6 Hz, 1H). 1.77 (s, 3H). di-tert-butyl (E)-(3-methylhept-2-en-6-yn-l-yl)-L-glutamate (S10): A procedure was adapted from the literature27as follows: To a solution of K2CO3(0.326 g, 2.36 mmol). KI (0.020 g, 0.118 mmol), and DMAP (0.014 g, 0.118 mmol) in acetonitrile (0.20 M) was added di-tert-butyl glutamate (0.349 g, 1 179 mmol), followed by S9 (0.221 g. 1. 179 mmol). After 48 hrs, the reaction mixture w as diluted with hexanes (60 mL), washed with H2O (3 x 30 mL), dried over Na2SO4and concentrated in vacuo. The residue was purified by silica gel chromatography (5: 1 hexanes:EtOAc) to give S10 (0. 123 g, 0.336 mmol. 28%). 41 NMR (500 MHz, D2O) δ 5.34 - 5.26 (m, 1 H), 3.26 (dd, J = 13.1, 6.7 Hz, 1H), 3.18 - .3.08 (m, 2H), 2.39 - 2.28 (m, 4H). 2.26 (d, J = 7.5 Hz, 2H), 1.98 (t, J = 2.5 Hz, 1H), 1.91 (dp, J = 13.3, 5.9 Hz, 1H), 1.82 (dq, J = 14.0, 7.3 Hz, 1H), 1.66 (s, 3H), 1.50 (d, J = 2.2 Hz, 9H), 1.46 (d, 2.3 Hz, 9H);13C NMR (500 MHz, D2O) δ

[0058] 174.45. 172.55. 123.54. 84.09, 81.26, 80.25, 68.56, 60.52, 45.30. 38.28, 32.08. 28.61 , 28.13. 28. 11, 17.34. 16.04.

[0059] (E)-(3-methy!hept-2-en-6-yn-l-yl)-L-glutamic acid (S11): A procedure was adapted from the literature27as follows: TFA (1.480 mL. 19.184 mmol) was added dropwise to a solution of S 10 (0. 123 g, 0.336 mmol) in CHCl3(1.65 mL) and stirred at room temperature for 18 hrs. The reaction mixture was washed with MeOH (2 x 5 mL) and solvent removed under reduced pressure to obtain the product Sil (0. 163 g, 0.341 mmol, quantitative yield) in TFA salt form.1H NMR (500 MHz, D2O) δ 5.29 (ddt, 8 9, 7.7, 1.5 Hz, 1H), 3.79 (dt, J = 8.5, 4.2 Hz, 1H), 3.76 - 3.67 (m, 2H), 3.64 (s, 1H), 2.59 - 2.45 (m, 2H), 2.37 - 2.31 (m, 3H), 2.24 (t, J = 6.9 Hz, 2H). 2.18 (dtd, .7 = 14.6, 7.3, 5.1 Hz. 1H), 2.06 (dtd. J = 14.5, 7.8, 6.6 Hz, 1H). 2.01 (s, 1H), 1.67 (d, J = 1.4 Hz, 3H);13C NMR (500 MHz. D2O) 113.83, 58.62. 52.37, 49.01, 43.84. 37.36, 29.91, 24.79, 20.75, 16.26. 15.58: HRMS (ESI) Calculated for C13H19NO4253.1314, found 252.1244 |M-H]-. (2E,6E)-9-(3,3-dimethyIoxiran-2-yl)-3,7-dimethylnona-2,6-dien-l-yl acetate (S5) Farnesyl epoxide mix: A procedure from the literature18was modified as follows: To a solution of farnesyl acetate (3.473 g, 13.136 mmol) in dry DCM (30 mL) at 0 °C under argon atmosphere was added 75% m-CPBA (3.117g, 14.449 mmol) in dry DCM (30 mL) dropwise via addition funnel over 7 mm. After stirring in an ice-water bath for 2 hrs, the reaction was quenched with 5% aqueous Na2S2O3, warmed to room temperature and diluted with saturated aqueous NaHCCL (50 mL). The organic layer was separated, aqueous layer extracted with DCM (2 x 50 mL), and pooled organic layers washed with brine, dried over MgSO4. and concentrated in vacuo. The crude residue (3 2 g) was adsorbed into 5 g of silica and dry loaded onto a 50 g SFAR Silica D 60 Biotage column that was pre-equilibrated with 5 CVs of 2% EtOAc in Hexanes. With a flow rate of 100 mL / min, hold 2 - 5 % B for 6 CV, hold 6 % B for 3,8 CV, hold 6 - 40 % B for 2.8 CV, hold 40 % B for 3.9 CV hold 40 - 80 % B for 1 CV. hold 40 % B for 1.5 CV. Fractions containing desired products were identified by UHPLC-MS, pooled, and concentrated in vacuo to afford recovered famesyl acetate (0.489 g, 14.1% yield), a mixture of desired monoepoxide S5 and monoepoxide S5b (1.636 g, 44.4% yield) and diepoxide S5c (0.805 g, 20.7% yield), each as a colorless oil.1H NMR (500 MHz. CDCl3) δ 5.41 - 5.35 (m, 1H), 5.33 (t, J = 7.3 Hz, 1H), 5.18 - 5.11 (m, 1H), 5.08 (q, J = 7.8 Hz, 1H), 4.58 (dd, J = 6 9, 3.3 Hz, 3H), 2 71 (dt, J = 12.8, 6.4 Hz, 2H), 2.25 - 2.07 (m, 5H), 2.06 (s, 1H), 2.04 (s, 7H), 1.73 - 1 .65 (m. 9H), 1.63 - 1.57 (m, 7H), 1.32 - 1 .22 (m, 9H).

[0060] (2E,6E)-3,7-dimethyl-10-oxodeca-2,6-dien-1-yl acetate (S6): A procedure from the literature18was modified as follows: To a solution of mixture of epoxides S5 and S5b in 10 mL THF and 5 mL of water was added NalO4(0.858 g, 3.210 mmol) and HIO42 H2O (1.341 g, 5.884 mmol) at 0 °C, and the mixture was stirred at the same temperature for 1 h. After cooling to room temperature, the reaction mixture was diluted with saturated aqueous NaHCO3(20 mL), extracted with EtOAc (3 x 30 mL) and pooled organic layers washed with brine, dried over MgSO4, filtered and concentrated in vacuo. Tire crude residue (1.4 g) was adsorbed into 5 g of silica and dry loaded onto a 50 g SFAR Silica D 60 Biotage column that was pre-equilibrated with 2 CVs of 5% EtOAc in Hexanes With a flow rate of 100 mL / min, hold 5 % B for 3 CV, hold 5 - 6 % B for 1.1 CV, hold 6% B for 1 CV, hold 6 - 10 % B for 3 8 CV, hold 10 - 20 % B for 5 CV, hold 20 % B for 5 CV, hold 20 50 % B for 1 CV, hold 50 % B for 1.7 CV to afford a clear colorless oil S6 (0.457 g, 1 .936 mmol) in 36% yield. R,- = 0.67 (3: 1 hexanes: EtOAc);1H NMR (500 MHz, CDCl3) δ 9.73 (t, J = 2.0 Hz, 1H), 5.33 - 5.27 (m, 1H). 5. 14 - 5.08 (m, 1H). 4,56 (d, J = 7. 1 Hz, 3H), 2.49 (td, J = 7.5, 1.9 Hz, 2H), 2.30 (t, J = 7.5 Hz, 2H), 2.10 (q. J = 7.3 Hz, 2H), 2.05 - 2.00 (m, 6H). 1 .68 (d, J = 1.4 Hz, 3H), 1.60 (d, J = 1 .4 Hz, 3H);13C NMR (500 MHz, CDCl3) δ 202.64, 171.19, 141.93, 133.57, 124.86, 119 45, 118.65, 61,44, 42.20, 39.37, 31.91 , 26.14, 21.13, 16.51 , 16.21; HRMS (ESI) calculated for C14H22O3238. 1569, found 297. 1619 [M + Hac - H]\

[0061] (2E,6E)-3,7-dimethylundeca-2,6-dien-10-yn-l-ol (S7): Compound S7 was prepared as follows17: To a round bottom flask charged with a stir bar and K2CO3(0.483 g. 3.495 mmol) was added a solution of S6 (0.417 g, 1.748 mmol) m HPLC grade MeOH (10 mL) and allowed to stir for 15 min. DAMP (0.403 g, 2.097 mmol) was added via syringe and stirred at room temperature for 18 hrs. The reaction mixture was diluted with Et2O (40 mL), washed with 5% aqueous NaHCO3(20 mL), brine (20 mL), dried over anhydrous Na2SOr, filtered and concentrated to afford a lightyellow oil (0.318 g, 1.655 mmol) in 95% yield. Rf= 0.60 (30% EtOAc in hexanes):1H NMR (500 MHz, CDCl3) δ 5.40 (t, J = 6.8 Hz, 1H), 5.17 ( t. .7 6.6 Hz, 1H), 4. 14 (d, .7 6.8 Hz, 2H), 2.26 (t. J = 7.3 Hz, 2H), 2.19 (t, J = 7.1 Hz, 2H), 2 12 (q, J = 6.9 Hz, 2H), 2 04 (t, J = 7.4 Hz, 2H), 1.94 (s, 1H), 1.67 (s, 3H), 1.60 (s, 3H).

[0062] (2E,6E)-3,7-dimethylundeca-2,6-dien-10-ynal (S8): A procedure was adapted from the literature72as follows: Dess Martin penodinane (0.803 g, 1.893 mmol) was added to a solution of S7 (0 280 g, 1 456 mmol) in dry CH2Cl2(10 mL) at once and stirred at room temperature for 1 h The reaction mixture was quenched with an aqueous 10% sodium thiosulfate solution (10 mL) and washed with water (10 mL). Organic compounds were extracted from pooled aqueous layers with CH2Cl2(3 x 20 mL). Pooled organic layers were washed with brine, dried over MgSO4, filtered and concentrated zfi vacuo. The crude reaction mixture was purified by silica flash chromatography (10: 1 hexanes: EtOAc), yielding S8 as a clear yellow oil (0.209 g, 1.100 mmol, 76%). Rf= 0.83 (30% EtOAc in hexanes); NMR (500 MHz, CDCl3) δ 9.98 (d, J -- 8.0 Hz, 1H), 5.88 (d, J = 7.9 Hz, 1H), 5. 15 (s, TH), 2.25 (d, J = 12.8 Hz. 6H), 2.20 (d, J = 7. 1 Hz, 2H), 2.16 (s, 3H), 1 .93 (s, 1H), 1.62 (s. 4H);13C NMR (500 MHz, CDCl3) δ 127.68, 123.93, 68.89, 40.46. 38.41. 26.28, 25.75. 17 41, 16.12.

[0063] ((6E)-3,7-diniethyliindeca-2,6-dien-10-yn-l-yI)-L-gIutamic acid (3): A procedure was adapted from the literature22as follows: A solution of S8 (0.180 g. 0.946 mmol) in methanol (2.5 mL) was added dropwise to an aqueous solution (2.5 mL) of L-glutamic acid (0.167 g, 1 .135 mmol) and sodium hydroxide (0.091 g, 2.27 mmol) and stirred for 4 hrs at room temperature. The reaction mixture was cooled to 0 °C, sodium borohydride (0.054 g, 1.419 mmol) was added at once, and the reaction mixture was stirred for an additional Ih. Formic acid was added until pH 4. and the reaction mixture was washed with toluene before concentration in vacuo. The crude material was then purified following Flash Procedure B. Fractions containing desired products were identified by UHPLC-MS, pooled, concentrated in vacuo and lyophilized, affording a white solid (0.067 g, 0.208 mmol, 22%, both regioisomers).1H NMR (500 MHz, D2O) δ 5.33 (tdd, J = 6.9. 3.4, 1.6 Hz, 1H). 5.26 - 5. 17 (m, 1H), 4.88 (s, 1H), 4.31 - 4.16 (m, 1H), 3.75 - 3.60 (m, 2H), 3.57 (td, J = 6.1, 2.5 Hz, 1H), 2.60 - 2.54 (m, 1H), 2.57 - 2.44 (m. 1H), 2.44 - 2.31 (m, 1H), 2.28 (tdd, J = 6.1, 2.8, 1.3 Hz, 2H), 2.28 - 2.21 (m, 1H), 2.21 (d, J = 1.5 Hz, 1H), 2.20 (dd, J = 2.8, 1.3 Hz, 2H), 2.21 - 2.16 (m, 2H), 2.15 (s, 2H), 2.18 - 2.09 (m, 2H), 2.12 - 2.02 (m, 1H), 1.85 (t, J = 1.3 Hz. 1H), 1.81 - 1.75 (m, 2H). 1 .70 (d, J = 1.2 Hz, 1H), 1.68 - 1.63 (m. 3H);13C NMR (500 MHz. D2O) δ 175.56. 171.41, 146.01, 145.73, 134.30, 133.90. 124.46, 124.20. 114.68, 113.86, 83.30, 68.37, 68.30, 60.63, 43 94, 43.98, 39.19, 38.18, 38.16, 30 17, 25.71, 25 28, 16.90, 15.36, 14.45.; HRMS (ESI) Calculated for C18H27NO4321.1940, found 302.1756 [M-H2O-H]-.

[0064] (5E,9E)-ll-bromo-5,9-diniethylundeca-5,9-dien-l-yne (S12): Compound S12 was prepared according to literature2': To a solution of S7 (0.300 g, 1 .561 mmol) in ether at 0 °C was added PBr3(0.211 g, 0.781 mmol) dropwise under argon atmosphere. After 2.5 hrs the reaction mixture was treated with aqueous NaHCO3(15 mL), and extracted with ether (3 x 15 mL). The combined organic extract was washed with brine, dried over NasSCH, and concentrated in vacuo to give S12 (0.351 g, 1.382 mmol) in 88% yield. Rf = 0.88 (5% EtOAc in hexanes).1H NMR (500 MHz, CDCl3) δ 5.42 (t, J = 6.6 Hz, 1H), 5.18 (t, J = 6.5 Hz, 1H), 4.15 (d, J = 6.9 Hz, 2H), 2.27 (td, J = 6.7, 2.2 Hz, 2H), 2.20 (t, J =- 73 Hz, 2H), 2.13 (q, J = 7.0 Hz, 2H), 2.08 - 2.02 (m, 2H), 1.95 (d, J= 2.5 Hz, 1H), 1.68 (s, 3H), 1.61 (s, 3H), 1.28 (s, 1H). di-tert-butyl ((2E,6E)-3,7-diniethylundeca-2,6-dien-10-yn-l-yl)-L-ghitaniate (S13): A procedure was adapted from the literature27as follows: To a solution of K2CO3(0.368 g, 2.66 mmol), KI (0.022 g, 1.332 mmol), and DMAP (0.016 g, 0.133 mmol) in acetonitrile (0.20 M) was added di-tert-butyl glutamate (0.394 g, 1 .332 mmol), followed by S12 (0.340 g, 1 .332 mmol). After 48 hrs, the reaction mixture was diluted with hexanes (60 mL), washed with H2O (3 x 30 mL), dried over Na2SO4and concentrated in vacuo. The residue was purified by silica gel chromatography (5: 1 hexanes:EtOAc) to give S14 (0.115 g, 0.031 mmol. 9%).1HNMR (500 MHz, D2O) 3 5.17 (dt, J = 17.7, 6.7 Hz, 4H), 3.29 (dd, J = 9.3, 6.2 Hz, 1H), 3 20 - 3.10 (m. 4H), 2.37 (ddd, J = 15.5, 8.6. 6.3 Hz, 1H), 2.32 - 2.26 (m, 4H). 2.22 (q, J = 8.9, 7.8 Hz, 5H), 2.13 (q, J = 7.4 Hz, 4H), 2.04 (t, J = 1.6 Hz, 4H), 1.95 (t, J = 2.6 Hz, 2H), 1.93 - 1.85 (m, 2H), 1.64 (d, J = 9.3 Hz, 12H), 1.50 (s, 9H), 1.46 (s, 9H);13C NMR (500 MHz. D2O) 173.01, 172.37. 137.84, 133.29. 125.45, 123.23, 84.37, 80.67. 79.94. 68.36, 61.36, 47.71 , 39.71 , 38.40. 32.41. 28.39, 28.15, 26.49, 24.90, 17.60, 16.32, 15.80.

[0065] ((2E,6E)-3,7-dimethylundeca-2,6-dien-10-yn-l-yl)-L-glutaniic acid (S14): A procedure was adapted from the literature27as follows: TFA (0. 114 mL, 1.478 mmol) was added dropwise to a solution of S14 (0.030 g, 0.065 mmol) m CHCl3(0 3 mL) and stirred at room temperature for 48 hrs. Solvent was removed under reduced pressure to obtain the product S14 in TFA salt form.

[0066] The embodiments illustrated and discussed in this specification are intended only to teach those skilled in the art the best way known to the inventors to make and use the invention. Nothing in this specification should be considered as limiting the scope of the present invention. All examples presented are representative and non-limiting. The above-described embodiments of the invention may be modified or varied, without, departing from the invention, as appreciated by those skilled in the art in light of the above teachings. It is therefore to be understood that, within the scope of the claims and their equivalents, the invention may be practiced otherwise than as specifically described. Amino Acid (Protein) Sequences: DsKabC (Digenea simplex) [SEQ ID NO.1] MTVSKYNGVASTFNGVARRFDFAPLEADKLNWYPRSALPPEIPAIDLNKVNTEEELAQFL VDIRKSGLFYIVDHGIPEEISIGCYNAFREFCNLPEEAREKYNTDESFKSGGYVPFKGTSIG GGNLFERQKDFVVKFFWRGPSVVNRSPNDRFAEFHDEHHRKTAELAEKIITTILKALKTR FPEFHPDELKDNINVRNMFFSNRIYPEAPPDDGEKADYRLVPHRDLSFITLANQVPANNG FKGLFILTGDGEKIPVPPIRNSYLVFIGQGLSYLTNKYLPAALHGVDFPDNTNFEGSERASL ISFYEPNDYMMPSKNINPLPEEIFEKSCTFYDDVGVGRAGTTYNYVRYKFHEGYYL GfKabC (Grateloupia filicina) [SEQ ID NO 2] MTSFDFVAKTFDFAPVKADELKWYPRSALPPEIPVIDLNNVNTEEELAQFLVDIRKSGLF YVVNHDIPEEISIQTYNEFREFCKLPEEARQKYNTDRCFFNGGYVPFKATSINGGNKNKQ QRDFVVKFFWRGPHVVNRSPNERFAKWHHQYHTRTAELGEKVMTTIAKALKTRFPDFD PAELEDNVNPHNMIFSNRIYPEFPPSEGEDAEYRLTPHRDISYITLVNQTPANNGFKALFIV TGDGERVYVPPIRNSYLVFIGQSLSYLTNKYLPAALHGVAFPDGNLEGCERASLVSFYEPY DRMVPSKNIKPTAEEIAPKSCSFYDSIGCDKTGTTFTDVRAKFHSGYYV ReKabC (Rhodophysema elegans) [SEQ ID NO 3] MAVYNGVSQAFDFAPLDADSLNWCPQSSLPPEIPAIDLGKVNTRAELAQFLIDIRRSGLFY VVNHGVPEELSIGCYNLFRRFCNIPEEERAKYSTDHHFVNGGYMPYKSSSIGKANMGKD QKDFVVKYFWRGPRVENRTPTAEFKKYHDEYHRRTSGVADTVIEKIMQALATRFPDFDP DEYKENTNSHHMFFSNRLYPENEPEKGENVEYRLVPHRDLSLVTLAHQIPADNGYQGLF VLTGDGKKVPVPPIRNSYLVFLGQALSYLTNKYLPAGLHGVDFPEKNSFEGSERSSLISFY EPHDRMMPSKALTPKEDEVFDRSCSFFDDIGVDTSGTTYMYVKNKFHEGYYL PpKabC (Palmaria palmata) [SEQ ID NO 4] MPVYNGTSQAFNFAPLHPDSLNWCPKSSLPPEIPVVDLSKLNSEAELAQFLVDIRKSGLF YVVNHGVPEELSIGCYNRFRQFCNIPEDQRAEYSTDHHFVNGGYMPYKSSSIGKANKGK SQKDFVVKYFWRGPRVENRSPNADFKSYHDEYHRRTADIANSVITKILQALTTRFPDFDP AEYKDNINSHHMFFSNRLYPDNEPEKGENVEYRLVPHRDLSFVTLAHQIPADNGYQGLF VFTGDGKKVCVPPIRNSYLVFMGQAMSYLTNKYLPAGLHGVDFPEKNCFEGSERSSLISF YEPHDHMMPSKALTPKDDEIFDRGCSFFDDIGVDKTGTTYMYVKNKFHEGYYL PmDabC (Pseudo-nitzschia multiseries) [SEQ ID NO 5] MTVAINNETVVLTPNEDDVQVNKGKTLETSFPPLKGDDLKWFPRSSLPAEIPAIDIGKVST KEELEQFLVDIRKSGLFYIVNHGVPEEVSINVYNAFREFISTTTEEERMKYYTDTHFQNG GYVPFQGSSIRGGNLGKPQKDHVVKYFWRGPEVINRTPNEKFTEAHNMHHTETFKVAE KVIRTIFKALKLRFPDFDPMEFENTINSKKMFFTNRIYPQAEPSDEEEITHRLVPHLDTSFIT LANQVPADNGFQGLFVETGDGKKVKVPGIRNSYLVFIGQSLSYLTKNYLPSALHGVDKP PSDLFEGSERSSLITFYEPAEIIIPSKNINPNPEETSESCPFFYDIGLTVNDPEGTTWDFVKNK FITGYYAD CaRadC1 [SEQ ID NO 6] MFTIKGTELNLDFNPLEVEKLNWYPRSSLPPQIPAIDLNKINTKEELGQFLDDIRKSGLFYI INHGIPEDVSMGCYNSFRKFCNLPEETRMKYNTDESFQNGGYMPFKGTSVGGGNKFTK NKDFVVKYFWRGPAVLNRSPDPDFQRNHDEHHKRTAELAEKITNTILKALRTRFPNFDPE ELNHNINPRNMFFSNRIYPEIPPEDGENAKFRLVPHRDLSFITLANQLPAQNGFKGLFIVTG DNEKVYVPPIRNSYLIFIGQGLSYLTNKFLPAALHGVEFPGKENFEGSERSSLISFYEPDDK MTPSMNINPSQDEIFEESCKFYDDVGADKNGTTFTYVKYKFLHGYY Nucleotide Sequences: DsKabC (Digenea simplex) [SEQ ID NO 7] ATGACAGTAAGTAAGTACAACGGTGTTGCGAGTACGTTCAACGGTGTTGCAAGGAGA TTCGACTTTGCCCCATTGGAGGCAGATAAATTGAATTGGTATCCCAGGTCAGCTCTTC CTCCAGAAATCCCTGCTATCGACCTGAATAAGGTCAATACTGAAGAAGAACTGGCTC AATTCCTGGTCGATATTCGCAAATCTGGGCTCTTTTATATTGTTGATCATGGCATTCCAG AGGAGATCTCTATAGGATGTTACAATGCATTCCGTGAGTTTTGCAACCTTCCCGAGGA AGCAAGAGAGAAGTACAACACAGATGAGTCCTTTAAAAGCGGTGGCTATGTTCCTTT CAAAGGCACGTCGATTGGTGGAGGGAACTTGTTTGAACGACAGAAGGATTTCGTTGT CAAATTCTTTTGGAGAGGACCAAGTGTTGTCAATAGGTCTCCTAATGATCGCTTTGCC GAATTCCATGATGAACATCATCGAAAGACAGCTGAGTTGGCCGAAAAAATCATCACT ACCATTTTGAAAGCCCTGAAGACACGCTTTCCAGAATTCCATCCTGACGAGCTAAAG GATAATATCAATGTTCGAAACATGTTTTTCAGTAATCGAATCTATCCAGAGGCTCCGCC AGATGATGGAGAAAAAGCTGACTACCGACTTGTTCCTCATCGAGATCTCAGTTTTATC ACTCTTGCAAATCAAGTTCCGGCTAACAATGGATTCAAGGGTCTTTTTATACTGACAG GTGATGGAGAAAAAATTCCTGTTCCTCCAATCCGGAACAGCTACTTGGTCTTCATCGG TCAAGGCCTCTCATATCTCACGAATAAGTATCTTCCTGCAGCGCTTCATGGTGTGGACT TTCCAGACAATACAAATTTTGAAGGAAGTGAAAGAGCCTCTCTGATCAGCTTCTACG AGCCAAACGATTACATGATGCCATCCAAGAACATTAACCCTTTACCTGAGGAGATATT TGAGAAAAGCTGTACGTTTTACGATGATGTCGGAGTAGGGAGGGCTGGTACAACATA TAATTACGTGAGGTACAAATTTCATGAAGGATACTACCTTTAA GfKabC (Grateloupia filicina) (codon optimized) [SEQ ID NO 8] ATGACTTCTTTCGATTTCGTCGCTAAGACTTTCGATTTTGCTCCCGTCAAAGCTGACG AGTTGAAGTGGTATCCACGTTCCGCGTTACCACCAGAAATTCCTGTAATTGATTTGAA CAACGTCAATACGGAGGAGGAGCTTGCACAGTTCCTGGTGGACATCCGCAAGTCTGG ACTTTTCTACGTTGTCAACCACGATATCCCGGAGGAAATCTCAATCCAGACGTACAAC GAATTTCGTGAGTTTTGTAAATTGCCAGAAGAGGCCCGCCAAAAATACAATACTGATC GCTGCTTCTTCAATGGGGGATATGTTCCATTCAAAGCCACGAGCATTAATGGGGGTAA TAAGAACAAACAGCAACGCGACTTTGTCGTGAAGTTTTTCTGGCGCGGCCCACACGT GGTCAATCGTTCCCCCAATGAACGCTTTGCCAAATGGCACCATCAGTATCATACACGC ACTGCTGAATTAGGTGAAAAGGTCATGACCACAATCGCAAAAGCTTTGAAGACGCGC TTTCCTGACTTCGACCCGGCCGAACTGGAGGACAATGTGAACCCTCACAACATGATT TTTTCTAACCGCATTTATCCGGAATTCCCACCCAGTGAAGGGGAGGATGCTGAGTATC GCCTGACCCCCCACCGTGATATTTCCTACATTACCCTTGTGAATCAGACTCCTGCCAAT AACGGATTCAAAGCATTATTTATCGTAACCGGGGACGGCGAACGCGTTTACGTCCCTC CTATCCGCAACTCTTACTTGGTATTCATCGGGCAGAGCTTGAGCTACCTTACCAATAAA TACCTTCCAGCGGCACTTCACGGGGTCGCTTTTCCGGACGGCAATCTTGAGGGTTGC GAGCGTGCCTCACTGGTGTCTTTCTATGAACCGTATGATCGCATGGTCCCATCGAAAA ATATCAAACCAACGGCAGAAGAGATCGCTCCAAAATCCTGCTCCTTCTATGATTCGAT TGGGTGTGACAAGACAGGAACTACCTTTACTGATGTTCGTGCAAAGTTTCACTCGGG GTACTACGTCTAA ReKabC (Rhodophysema elegans) [SEQ ID NO 9] ATGGCGGTGTACAATGGCGTCTCGCAGGCCTTCGACTTCGCGCCGCTCGACGCCGAC AGCCTCAACTGGTGCCCCCAGTCGTCCCTGCCGCCCGAGATCCCCGCCATCGACCTC GGCAAAGTGAACACCAGGGCCGAGCTCGCCCAGTTCCTGATCGACATCCGCAGGTCC GGCCTCTTCTACGTCGTCAACCACGGCGTGCCGGAAGAGCTCTCGATCGGCTGCTAC AACCTCTTTCGCCGCTTCTGCAACATCCCCGAGGAGGAGCGCGCCAAGTACAGCACC GACCATCATTTCGTCAACGGCGGCTACATGCCATACAAGAGCTCCTCCATCGGCAAGG CGAACATGGGCAAGGACCAGAAGGACTTTGTCGTCAAGTACTTCTGGAGGGGGCCC CGCGTGGAGAACAGGACGCCCACCGCCGAGTTTAAGAAGTATCACGATGAGTACCAT CGCAGGACTTCGGGCGTCGCAGACACGGTCATAGAGAAGATTATGCAGGCGCTGGCG ACGCGCTTTCCGGACTTCGACCCCGACGAGTACAAGGAGAACACGAACAGCCATCA CATGTTTTTCAGCAACAGGCTCTACCCCGAAAACGAGCCGGAGAAGGGGGAGAACG TCGAGTACCGCCTTGTTCCGCATCGCGACCTCAGTTTGGTGACGCTCGCGCACCAGAT CCCCGCAGACAATGGCTACCAGGGCCTGTTCGTTCTTACCGGCGACGGGAAGAAGGT GCCTGTGCCGCCTATCCGCAACAGCTATCTCGTCTTCCTGGGGCAGGCGTTGTCTTAT CTCACCAACAAGTATCTGCCTGCTGGATTGCATGGCGTGGACTTTCCGGAGAAGAAC TCCTTTGAGGGGAGCGAGCGGTCGTCACTTATTTCATTCTACGAGCCGCACGACCGC ATGATGCCGTCGAAGGCGCTCACGCCAAAAGAGGATGAAGTTTTTGATAGAAGCTGC TCCTTTTTCGATGATATTGGTGTTGATACGAGTGGCACGACGTATATGTACGTGAAAAA TAAGTTCCACGAGGGCTATTATCTATAG PpKabC (Palmaria palmata) (codon optimized) [SEQ ID NO 10] ATGCCTGTGTATAATGGTACGTCACAAGCATTCAATTTTGCGCCGCTGCACCCAGATA GTTTGAATTGGTGCCCAAAGTCGAGTCTTCCTCCAGAAATTCCTGTAGTGGATCTGTC AAAACTTAATTCTGAAGCCGAGCTGGCACAATTTCTTGTGGACATTCGTAAGAGCGG GTTGTTTTATGTAGTCAATCATGGTGTCCCGGAAGAACTTTCTATTGGGTGTTACAATC GCTTCCGTCAATTCTGTAACATTCCTGAGGACCAACGTGCGGAATACTCGACCGACCA TCATTTTGTAAACGGGGGCTACATGCCGTATAAGAGTTCCAGCATCGGCAAGGCAAAT AAAGGTAAATCTCAAAAGGACTTTGTGGTTAAATACTTCTGGCGTGGACCGCGCGTT GAGAACCGTAGTCCGGATGCTGATTTCAAGAGCTATCACGACGAATACCACCGTCGC ACAGCTGATATCGCTAACAGCGTGATTACTAAAATTCTTCAGGCCCTGACGACGCGTT TTCCTGACTTTGACCCTGCTGAATATAAAGATAACATTAATTCCCATCATATGTTCTTCT CGAACCGCCTTTACCCAGACAACGAACCTGAGAAAGGCGAGAATGTAGAATACCGTT TAGTACCGCACCGTGACCTGAGCTTCGTGACCCTTGCTCACCAAATCCCGGCGGACA ACGGTTACCAAGGCCTTTTCGTTTTTACCGGCGATGGGAAGAAGGTTTGCGTGCCTCC GATTCGTAACAGCTATCTTGTCTTCATGGGACAGGCAATGTCGTATCTGACGAATAAG TACTTACCTGCAGGGTTGCATGGCGTGGATTTTCCTGAAAAGAACTGTTTCGAGGGGT CTGAACGCTCCTCTCTTATCTCATTTTACGAACCTCACGATCATATGATGCCCAGCAAA GCACTTACACCTAAGGATGATGAGATCTTCGATCGTGGTTGTTCGTTCTTTGATGACAT CGGCGTGGACAAAACAGGGACGACCTATATGTATGTCAAGAACAAATTCCATGAAGG TTACTACCTGTAA PmDabC (Pseudo-nitzschia multiseries) [SEQ ID NO 11] ATGACTGTGGCAATAAATAACGAAACCGTTGTTTTGACCCCCAACGAGGATGATGTTC AAGTCAACAAGGGTAAAACTCTCGAAACATCGTTCCCACCTCTCAAGGGCGACGATT TAAAATGGTTCCCCAGATCATCTCTACCTGCTGAAATCCCTGCCATTGACATCGGCAA AGTCAGTACAAAGGAGGAGTTGGAACAGTTTTTGGTCGACATTCGCAAATCAGGACT TTTCTACATTGTCAACCATGGTGTCCCCGAAGAGGTCTCAATCAATGTTTATAATGCCT TCAGAGAGTTTATCTCCACCACCACCGAGGAAGAGAGAATGAAGTATTATACGGACA CTCATTTCCAGAATGGTGGATACGTTCCTTTCCAAGGCTCTTCTATCCGCGGAGGAAA CCTGGGCAAGCCGCAGAAAGATCACGTCGTGAAGTACTTCTGGAGAGGACCTGAGG TTATCAACAGGACTCCAAACGAGAAATTTACCGAAGCTCATAATATGCACCACACAGA GACCTTCAAAGTAGCGGAGAAGGTGATTAGAACCATTTTCAAGGCCCTCAAGCTTCG TTTCCCAGACTTCGACCCGATGGAGTTTGAAAACACCATCAACTCTAAGAAGATGTTC TTCACCAACCGCATCTATCCGCAGGCCGAACCGAGTGACGAGGAAGAAATCACCCAT CGTCTTGTTCCGCACTTGGATACCAGTTTCATCACTTTGGCGAATCAAGTTCCTGCGG ACAATGGCTTCCAGGGTCTTTTCGTTGAGACCGGAGATGGAAAAAAGGTCAAGGTAC CGGGGATCCGTAATAGCTATTTGGTCTTCATCGGCCAAAGTTTGTCTTACCTCACAAA GAACTACCTTCCATCCGCCCTTCATGGCGTGGACAAGCCACCTAGTGATTTGTTCGAA GGAAGCGAACGGTCCTCGTTGATCACCTTTTATGAACCCGCCGAGATCATTATTCCAT CAAAGAATATTAACCCCAACCCAGAAGAGACCTCTGAGTCGTGCCCTTTTTTCTATGA CATCGGGTTGACTGTGAACGATCCAGAGGGTACTACATGGGATTTTGTGAAGAACAA GTTCATCACCGGATACTACGCTGAT CaRadC1 (Chondria armata) (codon optimized) [SEQ ID NO 12] ATGTTCACCATTAAGGGTACCGAGTTAAATCTGGACTTCAACCCGTTAGAGGTCGAGA AACTGAACTGGTACCCGCGCTCTTCATTACCACCGCAGATACCCGCGATTGATTTGAA CAAAATCAACACCAAGGAAGAACTCGGCCAGTTCTTGGACGACATCCGGAAATCAG GGTTATTTTATATAATCAACCACGGTATTCCAGAGGACGTTTCAATGGGTTGTTACAAC AGCTTTAGAAAGTTTTGCAACCTGCCGGAAGAGACGCGAATGAAATATAATACTGAC GAGTCATTTCAGAATGGCGGTTATATGCCATTTAAGGGTACCTCTGTAGGCGGCGGCA ATAAATTCACTAAGAACAAGGACTTCGTAGTAAAGTACTTTTGGCGTGGGCCCGCAG TGTTGAACCGTAGCCCTGACCCTGACTTCCAGCGTAATCACGATGAGCACCACAAAA GAACCGCTGAGTTGGCGGAAAAGATTACTAACACCATCCTTAAAGCCCTTCGTACAC GCTTCCCTAACTTCGACCCAGAGGAGCTTAACCACAACATTAACCCACGCAACATGT TCTTTTCCAACCGGATATACCCTGAGATCCCACCCGAGGACGGTGAGAACGCGAAAT TCCGGCTGGTGCCCCATCGGGACCTGTCATTTATAACTCTGGCTAATCAACTGCCCGC CCAGAACGGGTTCAAGGGCCTCTTCATTGTTACCGGTGACAACGAGAAGGTCTACGT

[0067] CCCGCCCATTCGCAATAGCTATCTGATTTTCATAGGCCAGGGGCTGTCATACCTTACGA

[0068] ATAAGTTCCTGCCAGCGGCGTTACACGGCGTGGAGTTCCCTGGCAAAGAGAATTTCG

[0069] AGGGGTCTGAGAGAAGCAGTCTGATTrCCTTCTArGAACCCGArGATAAGAlGACACC

[0070] GAGCATGAATATAAATCCCAGCCAGGACGAAATCTTCGAGGAATCTTGCAAATTTTAT

[0071] GACGACGTCGGTGCTGATAAGAACGGAACAACCTTCACATATGTTAAGTATAAATTCC

[0072] TGCACGGATATTAITAA

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Claims

CLAIMSClaim 1. A compound of formulawherein n is at least 1 .Claim 2 . The compound of claim 1, wherein n is 1 or 2Claim 3, The compound of claim I, of formulaClaim 4. The compound of claim 1, of formulaClaim 5. A compound of formulawherein m is at least 0.Claim 6. The compound of claim 5, wherein m is 0 or 1.Claim 7. The compound of claim 5 of formulaClaim 8. The compound of claim 5 of formulaClaim 9. A compound of formulawherein m is at least 0 and wherein G is is selected from the group consisting of a fluorescent dye, a coumarin dye, coumarin, a triarylmethane dye, fluorescein, a cyanine dye, Cy5, a squaraine dye, SeTau-647, biotin, and benzyl.Claim 10. The compound of claim 9, wherein m is 0 or 1 . Claim 11. The compound of claim 9 of formulaClaim 12. The compound of claim 9 of formulaClaim 13. The compound of claim 9 of formulaClaim 14. The compound of claim 9 of formulaClaim 15. A method of preparing a compound of formulacomprising reacting a 1stcompound(1stcompound) with an epoxidizing agent to obtain a 2ndcompound(2ndcompound) reacting the 2ndcompound with an oxidizing agent to obtain a 3rdcompound(3rdcompound) reacting the 3rdcompound with a homologation agent, optionally a Seyferth -Gilbert reagent, dimethyl (diazomethyl)phosphonate. and / or dimethyl l-diazo-2-oxopropylphosphonate, to obtain a 4thcompound(4thcompound) reacting the 4thcompound with an oxidizing agent to obtain a 5thcompoundreacting the 5thcompound with a glutamic acid (Glu), wherein n is at least 1.Claim 16. The method of claim 15, wherein n is 1 or 2.Claim 17. The method of claim 15, comprising preparing a compound of formula(an alkynyl-functionalized prekainic acid) comprising reacting a 1stcompoundwith meta-chloroperoxybenzoic acid (mCPBA) in dichloromethane (DCM) to obtain a 2ndcompoundreacting the 2ndcompound with an alkali metal periodate in tetrahydrofuran (THF) and water (H2O) to obtain a 3rdcompound(3rdcompound) reacting the 3rdcompound with(compound A) and an alkali metal carbonate in methanol (MeOH) to obtain a 4thcompound(4thcompound) reacting the 4thcompound with Dess-Martin periodinane (DMP) in dichloromethane (DCM) to obtain a 5thcompound(5thcompound) reacting the 5thcompound with L-glutamic acid (L-Glu) in a solution of an alkali metal borohydride and an alkali metal hydroxide in methanol (MeOH) and water (H2O) to obtain the alkynyl-functionalized prekainic acid. Claim 18. The method of claim 17, wherein the alkali metal periodate is sodium periodate (NaIO4), wherein the alkali metal carbonate is potassium carbonate (K2CO3), wherein the alkali metal borohydride is sodium borohydride (NaBH4), and wherein the alkali metal hydroxide is sodium hydroxide (NaOH).Claim 19. A method of preparing a compound of formulacomprising transformingwith a kanoid synthase, wherein m is at least 0.Claim 20. The method of claim 19, wherein m is 0 or 1.Claim 21. The method of claim 19, wherein the kanoid synthase has at least 80%, 90%, 95%, 98%, or 99%, amino acid sequence identity to a KabC synthase, a DabC synthase, a RadCl synthase. DsKabC [SEQ ID NO. 1], GfKabC [SEQ ID NO. 2], ReKabC [SEQ ID NO. 3], PpKabC [SEQ ID NO. 4], PmDabC [SEQ ID NO. 5], or CnRadCl [SEQ ID NO. 6].Claim 22. The method of claim 19, wherein the kanoid synthase is selected from the group consisting of a KabC synthase, a DabC synthase, and a RadCl synthase.Claim 23. The method of claim 19, wherein the kanoid synthase is selected from the group consisting of DsKabC [SEQ ID NO. 1], GfKabC [SEQ ID NO. 2], ReKabC [SEQ ID NO. 3], PpKabC [SEQ ID NO. 4], PmDabC [SEQ ID NO. 5], and CaRadCl [SEQ ID NO. 6],Claim 24. The method of claim 19, comprising preparing a compound of formula(an alkynyl-functionalized kainic acid) comprising transforming(an alkynyl-functionalized prekainic acid) with CaRadC1 [SEQ ID NO. 6] protein in a solution of a-ketoglutaric acid (aKG), iron(II) sulfate (FeSO4), and L -ascorbic acid (L-asc) in water (H2O) to obtain the alkynyl-functionalized kainic acid.Claim 25. A method of preparing a compound of formulacomprising reactingwith an azide-functionalized group G, optionally in the presence of a copper salt, wherein m is at least 0. Claim 26. The method of claim 25, wherein m is 0 or 1Claim 27. The method of claim 25, comprising preparing a compound of formula(group G-functionalized triazole-functionalized kainic acid) comprising(an alkynyl-functionalized kainic acid) with an azide-functionalized group G, optionally in the presence of a copper salt, to obtain the group G-functionalized tri azol e-functionali zed kainic acid.Claim 28. The method of claim 27, wherein the group G is selected from the group consisting of a fluorescent dye, a coumarin dye, coumarin, a triarylmethane dye, fluorescein, a cyanine dye, Cv5, a squaraine dye, SeTau-647, biotin, and benzyl.Claim 29. The method of claim 27, wherein the copper salt is copper(II) sulfate (CuSO4).Claim 30. The method of claim 27, wherein the reacting is further in the presence of an alkali ascorbate and / or sodium ascorbate.Claim 31 . The method of claim 27, comprising preparing a compound of formula(coumarin-triazole functionalized kainic acid) comprising reacting(an alkynyl-functionalized kainic acid) with azide-functionalized coumarin (coumarin-Ns) in the presence of a copper salt to obtain the coumarin-triazole functionalized kainic acid.Claim 32. The method of claim 25, comprising preparing a compound of formula(benzyl-triazole functionalized kainic acid) comprising reacting(an alkynyl-functionalized kainic acid) with benzyl azide (Bn-N3) in the presence of a copper salt to obtain the benzyl-triazole functionalized kainic acid.

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