USOO9578881 B2

(12) United States Patent (10) Patent No.: US 9,578,881 B2 Zwiebel et al. (45) Date of Patent: Feb. 28, 2017

(54) COMPOSITIONS FOR INHIBITION OF A01N 29/02: A01N 33/18: A01N 33/22: INSECT SENSING A01N 47/08: A01N 47/14: A01N 53/00; A01N 57/12: A01N 57/14: A01N 57/16 (75) Inventors: Laurence Zwiebel, Nashville, TN (US); See application file for complete search history. Gregory M. Pask, Nashville, TN (US); (56) References Cited David C. Rinker, Nashville, TN (US); Ian M. Romaine, Nashville, TN (US); U.S. PATENT DOCUMENTS Gary A. Sulikowski, Brentwood, TN 2,798,053 A 7, 1957 Brown ...... 521.38 (US); Paul R. Reid, Durham, NC (US); 3,755,560 A 8, 1973 Dickert et al. 514,772.6 Alex G. Waterson, Murfreesboro, TN 4.418,534 A 12/1983 Dufft ...... 60,585 (US); Kwangho Kim, Nashville, TN 4.421,769 A 12/1983 Dixon et al. .... 514,772 (US); Patrick L. Jones, Danvers, MA 4,509,949 A 4, 1985 Huang et al...... 8,558 (US); Robert W. Taylor, Chicago, IL 4,599,379 A 7/1986 Flesher et al...... 524/801 4,628,078 A 12/1986 Glover et al. .. 526,303.1 (US) 4,835,206 A 5/1989 Farrar et al. .... 524/457 4,849,484 A 7, 1989 Heard ...... 525/221 (73) Assignee: Vanderbilt University, Nashville, TN 5,011,681 A 4, 1991 Ciotti et al...... 510, 136 (US) 5,087.445 A 2/1992 Haffey et al...... 424,59 5,100,660 A 3, 1992 Hawe et al...... 424/78.35 (*) Notice: Subject to any disclaimer, the term of this 5,567,430 A 10/1996 Levy ...... 424/409 5,698.210 A 12/1997 Levy ...... 424/406 patent is extended or adjusted under 35 5,824,328 A 10/1998 Levy ...... 424/409 U.S.C. 154(b) by 119 days. 5,846,553 A 12/1998 Levy ...... 424/409 5,858,384 A 1/1999 Levy ...... 424/406 (21) Appl. No.: 14/115,553 5,858,386 A 1/1999 Levy ...... 424/409 5,885,605 A 3/1999 Levy ...... 424/405 (22) PCT Fed: Apr. 25, 2012 5,902,596 A 5/1999 Levy ...... 424/405 (Continued) (86) PCT No.: PCT/US2O12/034847 S 371 (c)(1), FOREIGN PATENT DOCUMENTS (2), (4) Date: May 2, 2014 AU 2012254032 11 2013 CA 2835328 11 2012 (87) PCT Pub. No.: WO2O12A154403 (Continued) PCT Pub. Date: Nov. 15, 2012 OTHER PUBLICATIONS Prior Publication Data (65) Ex Parte Quayle Action issued on Jul. 29, 2015 for U.S. Appl. No. US 2014/0242135 A1 Aug. 28, 2014 13/881,638, filed Oct. 16, 2013 and published as U.S. 2014/ 0039013 on Feb. 6, 2014 (Inventor Zwiebel, et al.; Applicant— Vanderbilt University) (6 pages). Related U.S. Application Data Response to Ex Parte Quayle Action filed on Sep. 25, 2015 for U.S. Appl. No. 13/881,638, filed Oct. 16, 2013 and published as U.S. (60) Provisional application No. 61/483,440, filed on May 2014/0039013 on Feb. 6, 2014 (Inventor Zwiebel, et al.; Appli 6, 2011, provisional application No. 61/483,857, filed cant—Vanderbilt University) (5 pages). U.S. Appl. No. 61/406,368, filed Oct. 25, 2010, Zwiebel et al. on May 9, 2011, provisional application No. (Vanderbilt University). 61/540,929, filed on Sep. 29, 2011, provisional U.S. Appl. No. 61/406.786, filed Oct. 26, 2010, Zwiebel et al. application No. 61/586,492, filed on Jan. 13, 2012, (Vanderbilt University). provisional application No. 61/625,602, filed on Apr. U.S. Appl. No. 61/483,440, filed May 6, 2011, Zwiebel et al. 17, 2012. (Vanderbilt University). U.S. Appl. No. 61/483,857, filed May 9, 2011, Zwiebel et al. Int. C. (Vanderbilt University). (51) U.S. Appl. No. 61/540,929, filed Sep. 29, 2011, Zwiebel et al. AOIN 43/653 (2006.01) (Vanderbilt University). AOIN 43/78 (2006.01) AOIN 43/90 (2006.01) (Continued) AOIN 43/82 (2006.01) Primary Examiner — Blessing M Fubara AOIN 27/00 (2006.01) (74) Attorney, Agent, or Firm — Ballard Spahr LLP AOIN 29/02 (2006.01) (57) ABSTRACT AOIN 33/22 (2006.01) In one aspect, the invention relates to chemical modulators AOIN 53/00 (2006.01) of insect olfactory receptors. In particular, compounds and (52) U.S. C. compositions are provided that can inhibit sensory (e.g., host CPC ...... A0IN 43/653 (2013.01); A0IN 43/78 targeting) functions in airborne insects Such as mosquitos. (2013.01); A0IN 43/82 (2013.01); A0IN Method of employing Such agents, and articles incorporating 43/90 (2013.01) the same, are also provided. This abstract is intended as a (58) Field of Classification Search scanning tool for purposes of searching in the particular art CPC ...... A01N 43/653: A01N 43/78: A01N 43/90; and is not intended to be limiting of the present invention. A01N 43/82: A01N 25/34: A01N 27/00; 21 Claims, 40 Drawing Sheets

US 9,578,881 B2 Page 3

(56) References Cited Pitts et al. A highly conserved candidate chemoreceptor expressed in both olfactory and gustatory tissues in the malaria vector Anoph OTHER PUBLICATIONS eles gambiae, Proc. Natl. Acad. Sci. USA, 101:5058-5063, 2004. Qiu et al., Olfactory Coding in Antennal Neurons of the Malaria Jones et al., Functional conservation of an insect odorant receptor Mosquito, Anopheles gambiae, Chem. Senses, 31:845-863, 2006b. gene across 250 million years of evolution, Curr: Biol. 15:R119 Qiu et al., Attractiveness of MM-X Traps Baited with Human or R121, 2005. Synthetic, Odor to Mosquitoes (Diptera: Culicidae) in The Gambia, Jones et al., Two chemosensory receptors together mediate carbon Med. Vet. Entomol. 20:280-287, 2006a. dioxide detection in Drosophila, Nature, 445:86-90, 2007. Robertson and Wanner, Expansion of the odorant, but not gustatory, Kellogg, Water vapour and carbon dioxide receptors in Aedes receptor family. The chemoreceptor Superfamily in the honey bee, aegypt, J. 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Takken and Knols, Odor-mediated behavior of Afrotropical malaria Lindsay et al., Variation in attractiveness of human Subjects to mosquitoes, Annu. Rev. Entomol. 44:131-157, 1999. malaria mosquitoes (Diptera: Culicidae) in The Gambia, J. Med. Takken et al., Odor-mediated flight behavior of Anopheles gambiae Entomol. 30:308-373, 1993. gilessensu Stricto andAn. Stephensi liston in response to CO2, Liu et al., Distinct Olfactory Signaling Mechanisms in the Malaria acetone, and 1-octen-3-ol (Diptera: Culicidae), J. Insect Behavior, Vector Mosquito Anopheles gambiae, PLoS Biology 8(8): 10:395-407, 1997. e1000467, 2010. Takken, The role of olfaction in host-seeking of mosquitoes: a Lu et al., Odor Coding in the Maxillary Palp of the Malaria Vector review, Insect Sci. Appins., 12:287-295, 1991. Mosquito Anopheles gambiae, Curr: Biol. 17:1533-1544, 2007. Thomas, TCE: Biting activity of Anopheles gambiae, Brit. Med. J. Lundin et al., Membrane topology of the Drosophila OR83b odor 2: 1402, 1951. ant receptor, FEBS Lett., 581 (29):5601-5604, 2007. Vosshall et al. An olfactory sensory map in the fly brain, Cell, Mboera and Takken, Carbon dioxide chemotropism in mosquitoes 102: 147-159, 2000. (Diptera: Culicidae) and its potential in vector Surveillance and Vosshall et al., Aspatial map of the olfactory receptor expression in management programmes, Rev. Med. Vet. Entomol. 85:355-368, the Drosophila antenna, Cell, 96:725-736, 1999. 1997. Vosshall. The molecular logic of olfaction in Drosophila, Chem. Meijerink and van Loon, Sensitivities of antennal olfactory neurons Senses, 26:207-213, 2001. of the malaria mosquito, Anopheles gambiae, to carboxylic acids, J. Wetzel et al., Functional expression and characterization of a Insect Physiol. 45:365-373, 1999. Drosophila odorant receptor in a heterologous cell system, Proc. Meijerink et al., Olfactory receptors on the antennae of the malaria Natl. Acad. Sci. USA, 98:9377-9380, 2001. mosquito Anopheles gambiae are sensitive to ammonia and other Wicher et al., Drosophila odorant receptors are both ligand-gated Sweat-borne components, J. Insect Physiol. 47:455-464, 2001. and cyclicnucleotide-activated cation channels, Nature, Merrill et al., Molecular characterization of arrestin family members 452(7190): 1007-1011, 2008. in the malaria vector mosquito, Anopheles gambiae. Insect Molecul. Wistrand et al. A general model of G protein-coupled receptor Biol. 12:641-650, 2003. sequences and its application to detect remote homologs, Protein Merrill et al., Odorant-specific requirements for arrestin function in Sci., 15:509-521, 2006. Drosophila olfaction, J. Neurobiol. 63:15-28, 2005. Xia et al., The molecular and cellular basis of olfactory-driven Merrill et al., Visual arrestins in olfactory pathways of Drosophila behavior in Anopheles gambiae larvae, Proc. Natl. Acad. Sci. USA, and the malaria vector mosquito Anopheles gambiae, Proc. Natl. 105:6433-6438, 2008. Acad. Sci. USA, 99:1633-1638, 2002. Zwiebel and Takken, Olfactory regulation of mosquito-host inter Mombaerts, Molecular biology of odorant receptors in vertebrates, actions, Insect Biochem. Molec. Biol. 34:645-652, 2004. Annu. Rev. Neurosci., 22:487-509, 1999. Supplemental European Search Report issued May 28, 2014 for Muirhead-Thomson, Low Gametocyte Thresholds of Infection of European Patent Application No. 11838461.9, which was filed Oct. Anopheles with Plasmodium Falciparum, Brit. Med. J., I: 1114 21, 2011 and published as EP 2632257 on Sep. 4, 2013 (Inventor 1117, 1951. Zwiebel, et al.; Applicant Vanderbilt University) (pp. 1-9). Pelosi and Maida, Odorant-binding proteins in insects, Comp. International Preliminary Report on Patentability issued Apr. 30. Biochem. Physiol. B Biochem. Mol. Biol. 1 11:503-5 14, 1995. 2013 for International Patent Application No. PCT/US2011/057246, US 9,578,881 B2 Page 4

(56) References Cited International Preliminary Report on Patentability issued Mar. 25. 2014 for International Patent Application No. PCT/US2012/ OTHER PUBLICATIONS 034847, which was filed on Apr. 25, 2012 and published as WO 2012/154403 on Nov. 15, 2012 (Inventor Zwiebel, et al.; Appli which was filed on Oct. 21, 2011 and published as WO 2012/061039 cant Vanderbilt University) (pp. 1-8). on May 10, 2012 (Inventor—Zwiebel, et al.; Applicant Vanderbilt University) (pp. 1-7). International Search Report and Written Opinion issued Sep. 17. International Search Report and Written Opinion issued Jun. 21. 2012 for International Patent Application No. PCT/US2012/ 2012 for International Patent Application No. PCT/US2011/057246, 034847, which was filed on Apr. 25, 2012 and published as WO which was filed on Oct. 21, 2011 and published as WO 2012/061039 2012/154403 on Nov. 15, 2012 (Inventor Zwiebel, et al.; Appli on May 10, 2012 (Inventor—Zwiebel, et al.; Applicant Vanderbilt cant Vanderbilt University) (pp. 1-11). University) (pp. 1-11). Lee et al. "Preparation of triazole derivatives as T-type calcium Non-Final Office Action issued Dec. 15, 2014 for U.S. Appl. No. channel blockers.” Korea Institute of Science and Technology, 13/881,638, filed Oct. 16, 2013 and published as U.S. 2014/ (2008). 0039013 on Feb. 6, 2014 (Inventor—Zwiebel, et al.; Applicant— Vanderbilt University) (pp. 1-21). European Search Report issued May 13, 2015 for Application No. Response to Restriction Requirement filed Aug. 8, 2014 for U.S. 12781894.6 (Applicant Vanderbilt University) (8 pages). Appl. No. 13/881,638, filed Oct. 16, 2013 and published as U.S. U.S. Appl. No. 62/19 1960, filed Jul. 13, 2015, Zwiebel (Vanderbilt 2014/0039013 on Feb. 6, 2014 (Inventor Zwiebel, et al.; Appli Univ.) cant Vanderbilt University) (pp. 1-7). U.S. Appl. No. 62/138,348, filed Mar. 25, 2015., Zwiebel (Vanderbilt Restriction Requirement issued Jun. 9, 2014 for U.S. Appl. No. Univ.) 13/881,638, filed Oct. 16, 2013 and published as U.S. 2014/ International Search Report and Written Opinion issued on Jun. 20. 0039013 on Feb. 6, 2014 (Inventor—Zwiebel, et al.; Applicant— Vanderbilt University) (pp. 1-12). 2016 by the International Bureau for PCT/US2016/24076, filed on Preliminary Amendment filed Apr. 25, 2013 for U.S. Appl. No. Mar. 24, 2016 (Applicant Vanderbilt University; Inventors— 13/881,638, filed Oct. 16, 2013 and published as U.S. 2014/ Zwiewel et al.) (13 pages). 0039013 on Feb. 6, 2014 (Inventor—Zwiebel, et al.; Applicant— Vanderbilt University) (pp. 1-5). * cited by examiner U.S. Patent Feb. 28, 2017 Sheet 1 of 40 US 9,578,881 B2

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s R 9&oa So s ANSL) US 9,578,881 B2 1. 2 COMPOSITIONS FOR INHIBITION OF Disclosed are methods for disrupting insect odorant sens INSECT SENSING ing, the method comprising providing to an insect environ ment a compound that binds to and/or modulates insect Orco CROSS-REFERENCE TO RELATED ion channels. APPLICATIONS 5 Also disclosed are methods for mediating Orco response, the method comprising providing an effective amount of a This application claims the benefit of U.S. Provisional disclosed compound, or salt or tautomer thereof, to a Orco Application No. 61/483,440, filed on May 6, 2011; U.S. receptor, an Orco/ORX complex, or an Orco/Orco complex, Provisional Application No. 61/483,857, filed on May 9, wherein the compound binds and/or modulates the receptor 2011; U.S. Provisional Application No. 61/540,929, filed on 10 or complex. Sep. 29, 2011; U.S. Provisional Application No. 61/586,492, filed on Jan. 13, 2012; and U.S. Provisional Application No. Also disclosed are compositions comprising a compound 61/625,602, filed on Apr. 17, 2012, all of which are incor that binds to and/or modulates insect Orco ion channels, porated herein by reference in entirety. combined with a suitable carrier. 15 Also disclosed are articles comprising a compound that BACKGROUND binds to and/or modulates insect Orco ion channels. Also disclosed are compounds having a structure repre Olfaction plays a critical role in insect behaviors among sented by a formula: agricultural pests and disease vectors. Hildebrand, et al., 1997, Annu. Rev. Neurosci, 20:595-631. Insect behavior is largely directed by the sensation of environmental olfactory cues (Gilliot C (2005) Entomology. 3rd Edition). The ability of an insect to respond to chemical stimuli is necessary for the insect to reproduce, mate, and feed. For example, insects 25 respond to certain chemical stimuli by moving up a chemical gradient to identify and target a host. R1 This behavior contributes to the spread of diseases in humans, such as malaria, encephalitis, and dengue fever, as well as, animal and livestock diseases and can result in 30 severe crop damage. More important to human health, the destructive behaviors of disease vector mosquitoes and related dipterans are driven by the sensory modality of wherein: R is hydrogen or is taken together with R to be olfaction, making it an important area of study (Carey AF. alkanediylcar alkenediylcar or a substituted version of Carlson J R (2011) Proc Natl Acad Sci U.S.A. 108: 12987 35 either of these groups; R is hydrogen or is taken together 12995). Mosquitoes, in particular, are believed to use olfac with R" as defined above; and R is hydrogen, hydroxy, tion to identify and target sources of bloodmeal for repro nitro, halo, alkylos. Substituted alkylos, alkenylcs, or ductive purposes. substituted alkenyls: or a salt or tautomer of the formula. The primary tool against insect borne diseases and crop damage due to insects is the use of insecticides that kill or 40 Also disclosed are compounds of the formula: repel the insect. However, each of the various forms of insecticide treatment—residual house spraying, crop dust ing, insecticide treated clothes, bedding and netting, and chemical larviciding have drawbacks, including environ mental and host toxicity, limited duration and need for insect 45 contact. Biological larviciding can avoid toxicity issues, but takes time and is quite expensive. Chemoprophylaxis is also expensive and may have unacceptable side effects. Finally, segregating populations is expensive and in many cases (third world countries) impractical. 50 Thus, while there are many different ways to attack insect pests, and each have contributed Substantially to limiting the spread of disease and/or crop damage, they also each have limitations that leave room for Substantial improvement. wherein: R is hydrogen or is taken together with R to be Despite advances in the field, there is still a scarcity of 55 alkanediylcar alkenediyl or a substituted version of compounds that inhibit insect sensing. This need and other either of these groups; R2 is hydrogen, alkyls, substituted needs are satisfied by the present invention. alkyls, or is taken together with R as defined above. Rs is hydrogen, hydroxy, nitro, halo, alkyls substituted SUMMARY alkylos), alkenylcs), or Substituted alkenyls). R4 is 60 In accordance with the purpose(s) of the invention, as alkylcs): alkenylcs arylc-10): aralkylcio. embodied and broadly described herein, the invention, in heteroarylcs, heteroaralkylcs, or substituted versions of one aspect, relates to entomology and infectious disease. any of these groups; Rs is heteroaryl- or substituted More particular, the invention relates to methods and com heteroarylco; and R is hydrogen, alkylcs, substituted positions for disrupting olfactory processes that underly 65 alkylcs, alkenyls, or substituted alkenylcs, or a salt many critical behaviors (e.g., host-targeting) in insects (e.g., or tautomer of the formula; provided that if R and R are H mosquitoes). and Rs is 3-pyridinyl, then R cannot be ethyl. US 9,578,881 B2 3 4 Also disclosed are compounds of the formula: or R' is taken together with a substituent of R to form a five-, six-, or seven-membered heterocylcoalkyl ring; wherein R is optionally substituted and selected from (C1 N-N C5) alkyl, (C1-C5) alkenyl, (C6-C10) aryl, (sC10) aralkyl, R11 X (sC8) heteroaryl, and (sC8) heteroaralkyl; wherein R is X^n N O optionally substituted aryl or optionally substituted (sC6) ? V heteroaryl; and wherein R* and R' are independently N R4 selected from hydrogen, optionally substituted (C1-C5) 2 N alkyl, or optionally substituted (C1-C5) alkenyl, or R'' and R1 10 R', along with the intermediate carbon, together comprise a C3-C6 cycloalkyl ring or a C2-C5 heterocylcoalkyl ring: or a salt or tautomer thereof, wherein the compound is not: wherein: R is hydrogen or is taken together with R to be alkanediyl alkenediyl or a substituted version of 15 either of these groups; R2 is hydrogen, alkyls substituted alkyls, or is taken together with R as defined above: Rs is hydrogen, hydroxy, nitro, halo, alkyls, substituted alkylcs, alkenyles, or Substituted alkenyles: R is O alkylc-s) alkenylcs arylc-10): aralkylcio. heteroarylcs, heteroaralkylcs, or substituted versions of any of these groups; and R'' is H. -OH, - F. —Cl, Br, I, NH, NO. —COH, -COCH, —CN, SH, —OCH, OCHCH. —C(O)CH, N(CH), —C(O) NH, —OC(O)CH, or—S(O)NH2 or a salt or tautomer of 25 the formula, wherein the compound is not: Also disclosed are methods for preparing a compound, the method comprising the steps of providing a compound 30 having a structure represented by a formula: C-terro O 35 Also disclosed are compounds having a structure repre --- sented by a formula: H

wherein R is optionally substituted aryl or optionally sub 40 stituted (

45 R4 wherein m, n, p, and q are independently 0 or 1; wherein L' R N and L are independently divalent organic groups having y - Q from 1 to 8 non-hydrogen members; wherein Q' is —O , N-N S-, -S(O)—, or -S(O) ; wherein Q is -O-, 50 —S , or NR'; wherein R is optionally substituted and selected from monocyclic aryl, bicyclic aryl, monocyclic wherein Q' is —O— or - S ; wherein R is optionally heteroaryl, bicyclic heteroaryl, and tricyclic heteroaryl; substituted and selected from (C1-C5) alkyl, (C1-C5) alk wherein R' is hydrogen, optionally substituted C1-C4 alkyl, enyl, (C6-C10) aryl, (sC10) aralkyl, (sC8) heteroaryl, and optionally substituted phenyl, optionally substituted benzyl, 55 (sC8) heteroaralkyl. or a structure represented by a formula selected from: Also disclosed are methods for preparing a compound, the method comprising the steps of providing a compound having a structure represented by a formula: O O A-s- a-- R4 O HN

A-. 65 {rs,N US 9,578,881 B2 5 6 wherein Q' is - O - or-S , and wherein R is optionally wherein Q' is —O— or - S ; wherein R is optionally substituted and selected from (C1-C5) alkyl, (C1-C5) alk substituted and selected from (C1-C5) alkyl, (C1-C5) alk enyl, (C6-C10) aryl, (sC10) aralkyl, (sC8) heteroaryl, and enyl, (C6-C10) aryl, (sC10) aralkyl, (sC8) heteroaryl, and (sC8) heteroaralkyl, reacting with a compound having a (

O XI O R7 10 N s 7 R6a R6b

R1 pi

pi wherein X" is a leaving group; wherein n is 0 or 1; wherein 15 R’ is optionally substituted (C6-C10) aryl or optionally While aspects of the present invention can be described substituted (animals. While males typi being square, but later in development they become hex cally feed on nectar and plant juices, the female needs to agonal. The hexagonal pattern will only become visible obtain nutrients from a “blood meal' before she can produce when the carapace of the stage with square eyes is molted. eggS. The head also has an elongated, forward-projecting 'stinger Mosquito larvae have a well-developed head with mouth 65 like' proboscis used for feeding, and two sensory palps. The brushes used for feeding, a large thorax with no legs and a maxillary palps of the males are longer than their proboscis segmented abdomen. Larvae breathe through spiracles whereas the females' maxillary palps are much shorter. As US 9,578,881 B2 33 34 with many members of the mosquito family, the female is ornamental plants, including flowers, and trees which may equipped with an elongated proboscis that she uses to collect be targeted using the methods and compositions of the blood to feed her eggs. present invention. The thorax is specialized for locomotion. Three pairs of Termites. Odontotermes obesus Rambur and Microtermes legs and a pair of wings are attached to the thorax. The insect 5 obesi Holmgren. Social insects that live underground in wing is an outgrowth of the exoskeleton. The Anopheles colonies; attack young seedlings as well as grownup plants; mosquito can fly for up to four hours continuously at 1 to 2 the attacked plants rather wither and ultimately die. kilometres per hour (0.62 to 1.2 mph) travelling up to 12 km Stem-borer. Sesamia inferens Walker. Moths are straw (7.5 ml) in a night. coloured, lay eggs in clusters inside the leaf-sheaths; pink The abdomen is specialized for food digestion and egg 10 ish-brown caterpillars bore into stems and kill central development. This segmented body part expands consider shoots; causing dead-hearts ably when a female takes a blood meal. The blood is Gujhia weevil. Tanymecus indius Faust. Adults are earth digested over time serving as a Source of protein for the ern-grey weevils, grubs feed on roots, whereas the adults cut production of eggs, which gradually fill the abdomen. 15 growing-points or nibble at margins of leaves; severer at the In order for the mosquito to obtain a blood meal it must Seeding stage. circumvent the vertebrate physiological responses. The mos Cutworms. Agrotis ipsilon Hufner and A. flammantra quito, as with all blood-feeding arthropods, has mechanisms Schiffer-Mueller. Caterpillars are general feeders. to effectively block the hemostasis system with their saliva, Thrip. Anaphothrips flavinctus Karny. Nymphs and adults which contains a mixture of Secreted proteins. Mosquito lacerate tender leaves, causing characteristics whitish saliva negatively affects vascular constriction, blood clot streaks; low temprature favourable to rapid multiplication. ting, platelet aggregation, angiogenesis and immunity and Wheat aphids. Schizaphis (Toxoptera) graminum Ron creates inflammation. Universally, hematophagous arthro dani, Rhopalosiphum maidis Fitch and Sitobion avenae pod saliva contains at least one anticlotting, one anti Fabricius. Nymphs and adults Suck sap from leaves, tender platelet, and one vasodilatory Substance. Mosquito Saliva 25 shoots and immature grain; multiply extremely fast, forming also contains enzymes that aid in Sugar feeding and antimi large colonies. crobial agents to control bacterial growth in the Sugar meal. Surface grasshopper. Chrotogonus trachypterus The composition of mosquito saliva is relatively simple as it Blanchard. Adults stout, mud-like in colour, polyphagous, usually contains fewer than 20 dominant proteins. Despite feeding on foilage and tender shoots. the great Strides in knowledge of these molecules and their 30 Shoot fly. Atherigona naq vii Steyskal. The fly has role in bloodfeeding achieved recently, scientists still cannot assumed the status of a pest recently; maggots attack seed ascribe functions to more than half of the molecules found lings and kill the central shoots, causing dead-hearts. in arthropod saliva. One promising application is the devel Galerucid beetle. Madurasia obscurella Jacoby. Adult opment of anti-clotting drugs based on saliva molecules, beetles feed on foilage and make small circular holes in the which might be useful for approaching heart-related disease, 35 leaves; active during July–October. because they are more user-friendly blood clotting inhibitors Jassid. Empoasca herri Pruthi. Nymphs and adults remain and capillary dilators. on the underside of the leaves and Suck the sap, leaves turn Two important events in the life of female mosquitoes are brown and crumple. egg development and blood digestion. After taking a blood 40 Plume moth borer. Exelastis atomosa Walsingham. A meal the midgut of the female synthesizes proteolytic specific pest of red-gram; slender buff-colored moths, hav enzymes that hydrolyze the blood proteins into free amino ing plumose wings; greenish-brown hairy caterpillars feed acids. These are used as building blocks for the synthesis of on flowers and later on bore into pods to feed on the egg yolk proteins. developing seeds inside. b. Other Insect Disease Vectors 45 Gram pod fly. Agromyza obtusa Mallas. A serious pest of In addition to mosquitoes, the inventors contemplate red-grain; the Small met allic-black fly lays eggs on pods; application of the compounds and methods of the present maggots bore into the pods and feed on the seeds; occasion invention against other insect disease vectors, including ally early in the season, grubs mine leaves. those that promote non-human disease. For example, aphids Hairy caterpillars. Ansacta moorei Butlei, Albistriga are the vectors of many viral diseases in plants. Fleas (Such 50 Walker, Diacrisia obliqua Walker, Euproctis fraterna as the human flea, Pulex irritans, and the oriental rat flea, Moore, E. scintillans Walker Polyphagous. Caterpillars feed Xenopsylla cheopis) transmit bubonic plague, murine typhus gregariously and Voraciously on foliage. and tapeworms. The glassy-winged sharpshooter transmits Cowpea stem fly. Melangromyza phaseoli Coquillett. A the Xylella fastidiosa bacterium among plants, resulting in small blue-black fly, thrusts eggs into the epedermis of soft diseases of grapes, almonds, and many other cultivated 55 stems; pale-yellow maggots after mining leaves travel plants. Phlebotomine sand flies transmit leishmaniasis, bar towards stem through the petiole and kill the young plants; tonellosis, sandfly fever and pappataci fever. Ticks of the the vigour of old plants is adversely affected. genus Ixodes are vectors of Lyme disease and babesiosis, Aphids. Aphis craccivora Kochi and A. cardui L. Colo and along with lice, transmit various members of the bac nies of nymphs and adults infest the tender growing shoots, terial genus Rickettsia. Triatomine bugs such as Rhodnius 60 flowers and young pods and Suck the Sap; infested parts dry prolixus are vectors of Chagas disease. Several genera of and no pod or seed formation takes place. Tsetse flies are vectors of human African trypanosomiasis Whitefly. Bemisia tabaci Gennadius. The flies suck the (also known as “African sleeping sickness'). sap from leaves and tender growing parts, which dry and c. Agricultural Pests wither. They act as the vector of yellow mosaic of legumes. The following is a list of agricultural pests for crops such 65 Sphinx moth. Agrius convolvuli Linnaeus. Stout dark as wheat, barley, oats, jowar, nuts, maize, soybean, Sorghum, brown moth; horned caterpillars defoliate plants by feeding pea, potato, cucumber, tomato, grams, rabi, rice fruits, Voraciously. US 9,578,881 B2 35 36 Leaf caterpillars. Azazia rubicans Biosduval. Sporadic; cosmopolitan; the tiny pinkish fly lay eggs inside the glumes the adult moth resembles a dry leaf green caterpillars feed and the larvae feed on the ovaries, thus preventing seed on leaves and tender plant parts. formation. Gram pod borer. Helicoverpa (Heliothis) obsoleta Fab Aphids. Phopalosiphum maidis Fitch and Aphis sacchari ricius Polyphagous. Moth yellowish brown; caterpillar 5 Zehntner. Nymphs and adults suck the sap from the leaves green, with dark broken grey lines, feed on foilage, later on and shoots, exclude honeydew, on which a Sooty mould bore into pods and feed on the seeds within. grows, giving the leaves a black appearance and interfering Gram caterpillars. Helicoverpa (Heliothis) armigera with photosynthesis. Hubner and H. zea, Boddie (obsoleta Fabricius). Polypha Deccan wingless grasshopper/Boliver Phadka grasshop gous; moths stout, light brown; caterpillars yellowish, make 10 per. Colemania sphenaroides/Hieroglyphus Bolivar. Eggs holes in pods and feed on the seeds within. are laid in the soil 75-200 mm deep; hoppers and adults feed Other pod borers. Etiella zinckenella Treitdche. Adult, on foilage, at times causing severe defoliation of the crops; greyish brown, with a distinct pale white band along the adults of C. sphenaroides are wingless, whereas those of H. front margin of the forewings; tiny greenish caterpillars, 15 nigrorepletus are short winged and can fly short distances with 5 black spots on the prothoracic shield, enter the pods only. and eat the seeds; more serious on green pea, specially in Earhead bug. Calocoris angustatus Lethierry. Nymphs nothern India. Adisura athinsoni Moore. A serious pest in and adult bugs suck the sap from tender grains at the milky Karnataka; moths pale-yellowish brown; the brownish stage, making them chafly. green caterpillars feed on the seeds by boring into the Sorghum shoot bug. Peregrinus maidis Ashmead. ripening pods. Maruca testutalis Geyer. A minor pest; adults Nymphs and adult bugs. Suck the sap from the leaves and with fuscous forewings, having transverse white markings; whorls, which turn pale green. pale-brownish caterpillars bore into the pods of various Hairy caterpillars. Ansacta moorei Butler, Estigmene pulses (kharif pulses as well) to eat seeds inside lactinae Cramer. General feeders, frequently causing severe Cut worms. Agrotis psilon Hubner, A. flammatra Schiffer 25 defoliation; caterpillars of A. moorei are red whereas those Mueller, A. Segetum Schiffer-Mueiller, A. spinifere Hubner. of E. lactinae are black. Aphids. Aphis crassivora Koch, A. medicagenis Koch and Earhead caterpillars. Eublemma (Heliothis) armigera Macrosiphum pisi Hubner Polyphagous. Nocturnal, stout Hubner and other species. Occur throughout the country; larvae, feed on leaves of young plants and cut the older ones caterpillers feed on maturing grains. at the ground level. Colonies of nymphs and adults attack 30 Mites. Oligonychus indicus Hirst and Schizotetranychus tender shoots, flowers and young pods and Suck the Sap: andropogoni Hirst. Colonies of nymphs and adults Suck the infested parts dry up. A. medicagenis is black, whereas M. sap from the undersurface of the leaves, causing reddish pisi is green, and A. crassivora is brownish. brown spots and patches. Pea leaf-miner. Phytomza atzicornis Meigen. A major pest Blister beetles. Lytta tenuicollis Pallasi and Zonabris of pea; polyphagous; maggots make ZigZag mines in the 35 pustulata Thunberg. Adult beetles feed on pollen and flow leaves; eat green matter and pupate inside; infected leaves CS. become whitish and dry up. Leaf roller. Marasmia trapezalis Guenee. Slender, yel Pea stem fly. Melanagromyza phaseoli Coquillett. A lowish-green caterpillars fold and roll the leaves near the major pest of pea, it also attacks kharif pulses; maggots tips and feed inside on the chlorophyll. attack young seeds inside the pods. The same as for the gram 40 Shoot fly. Atherigone approximata Malloch. The flies cut podd borer. the growing-points, causing dead-hearts during the seedling Pea semi-loopers. Plasia Orichalcea Fabricius and P stage, whereas in the advanced stage; they feed on earheads nigrisigna Walker. Polyphagous; moths with a golden patch and cut down peduncles. on the forewings (P. Orichalces); green caterpillars feed on Bajra midge. Geromyia pennisetti Harris. The larvae leaves during December to March. 45 destroy the ovaries seriously, affecting the development of Blue butterfly. Cosmolyee baeticus. Short pale-green cat seeds. erpillers feed on the leaves, flowers and pods of pea. Ragi white borer. Saluria inficita Walker. A specific pest Lucerne caterpillar. Laphygma exigua Hubner. Occasion of ragi; creamy white caterpillars bore into the stems close ally a serious pest of pea; dark-brown moths lay eggs on the to the soil surface; adults are dark brown, with a pale-white lower portion of the young plants; caterpillars feed on the 50 band along the margin of each forewing. leaves. Black hairy caterpillar. Estigmene exigua Hubner. Also Stem-borer beetles. Oberea brevis Gahan Nupserha known as woolly bear caterpiller; feed on leaves and ear bicolor Thomson. Pale brown longicorn beetles; grubs bore heads; the adults are creamy white moths with characteristic into the stems of growing plants. crimson marks on the head and the body. Gray weevils. Myllocerus spp. Adults feed on leaves, 55 Lucerne caterpillar. Spcdoptera exigua Hubner. Smooth, nibbling the leaf margins in the initial stage. brownish-green caterpillers feed on foilage, moving in large Shoot fly. Atherigona soccata Rodani. Damage caused numbers from field to field; common in nurseries. during the early seeding stage, larvae cut the growing points, Ragi-root aphid. Tetraneura hirsuta Baker. Minute, pale causing dead-hearts; tillers do develop after the central shoot white insect, found damaging roots, resulting in a gradual is killed, but the yield from these tillers is rather poor; 60 drying up of plants; infestation by the presence of black ants. commoner is early-sown rabi or late-sown kharif crops. Ragi jassid. Cicadulina bipunctella bipunctella. Nymphs Stem borers. Chilo zonellus (partellus) Swinhoe Ragi and and adults suck the sap from the leaves and stems; an Sesamia inferens Walker. Moth, dirty brownish, nocturnal, important vector of ragi mosaic virus. caterpillars feed on foilage and bore into the stems, causing Almond weevil. Myllocerus laetivirens Marshall; Mylo dead-hearts; also tunnel the stem and bore into earheads. 65 cerus undecimpustulatus Faust and M. discolor Boheman Sorghum midge. Contarinia sorghicola Coquillett. The Amblyrrhinus poricollis Boheman. Polyphagous pest, young insect has assumed the status of a serious pest recently: weevils feed on roots, whereas the adult weevils feed on the

US 9,578,881 B2 39 40 Paddy jassids. Nephotettix apicalis Motschulsky and N. vector control aimed at mosquito eradication, disease pre impicticeps Fabricius. Adults Small, green, with black spots vention, using prophylactic drugs and developing vaccines on forewings; nymphs and adults Suck plant sap; affected and prevention of mosquito bites, with insecticides, nets and plants turn yellow and growth is adversely affected. repellents. Since most such diseases are carried by “elderly White leafhoppers. Tettigella spectra Distant. Adults 5 females, scientists have Suggested focusing on these to avoid larger than those of Nephotettix spp. and white; both nymphs the evolution of resistance. and adults suck sap from young leaves; infested leaves turn a. Protozoa yellow. The mosquito genus Anopheles carries the malaria para Fulgorid bug. Nilaparvartha lugens Stal. Minor pest: site (see Plasmodium). Worldwide, malaria is a leading recorded feeding or ripening ear-heads. 10 cause of premature mortality, particularly in children under Paddy thrip. Cloethrips oryzae Williams. Nymphs and the age of five. It is widespread in tropical and Subtropical adult lacerate tissues; affected leaves present yellowish regions, including parts of the Americas (22 countries), Asia, streaks; tips curl and wither. and Africa. Each year, there are approximately 350-500 Whorl maggot. Hydrellia sp. Minor pest, common during million cases of malaria, killing between one and three kharif, maggots feed in the worls of developing leaves. 15 million people, the majority of whom are young children in Paddy mealy bug. Ripersia Oryzae Green. Colonies of sub-Saharan Africa. Ninety percent of malaria-related deaths reddish-white Soft insects infest Succelent paddy stems, occur in Sub-Saharan Africa. Malaria is commonly associ hidden by outer leaf-sheaths, Suck cell sap; growth gets ated with poverty, and can indeed be a cause of poverty and stunted; affects ear-head formation. a major hindrance to economic development. Rice root aphid. Tetraneura hirsuta Baker. Colonies of Five species of the Plasmodium parasite can infect nymphs and adults suck sap from roots just below soil humans; the most serious forms of the disease are caused by Surface, affected plants become pale and wither. Plasmodium falciparum. Malaria caused by Plasmodium Paddy leaf-roller. Cnaphalocrocis medinalis Guenee. vivax, Plasmodium ovale and Plasmodium malariae causes Sporadic pest: caterpillars roll the leaf tips and feed inside. milder disease in humans that is not generally fatal. A fifth Paddy skippers. Pelopides mathias Fabricius. Adult, a 25 species, Plasmodium knowlesi, is a Zoonosis that causes dark-brown butterfly: caterpillar, Smooth and green, feeds on malaria in macaques but can also infect humans. leaves. Malaria is naturally transmitted by the bite of a female Paddy root weevil. Echinocnemus Oryzae Marshal. Small Anopheles mosquito. When a mosquito bites an infected grey weevil, grubs attack paddy roots and affect the growth person, a small amount of blood is taken, which contains of plants. 30 malaria parasites. These develop within the mosquito, and Other pests include the Asiatic Garden Beetle, Asparagus about one week later, when the mosquito takes its next blood Beetles, Bean Leaf Beetle, Beet Webworm, Bluegrass Bill meal, the parasites are injected with the mosquito's saliva bug, Brown Marmorated Stink Bug, Cabbage and Seedcorn into the person being bitten. After a period of between two Maggot, Cabbage Looper, Cabbage Webworm, Carpenter weeks and several months (occasionally years) spent in the Ant, Carpenter Bee, Carpet Beetles, Catalpa Sphinx Cater 35 liver, the malaria parasites start to multiply within red blood pillar, Celery Leaftier, Cereal Leaf Beetle, European Corn cells, causing symptoms that include fever, and headache. In Borer, Click Beetle, Colorado Potato Beetle, Confused Flour severe cases the disease worsens, leading to hallucinations, Beetle, Corn Earworm, Cucumber Beetle, Cutworms, Dia coma, and death. mondback Moth, Eggplant Lace Bug, Flea Beetles, Fungus A wide variety of antimalarial drugs are available to treat Gnat, Green Peach Aphid, Hornworms, Hunting Billbug, 40 malaria. In the last 5 years, treatment of P falciparum Imported Cabbageworm, Indian Meal Moth, Japanese infections in endemic countries has been transformed by the Beetle, Lace Bugs, Leaf-Footed Bugs, Mexican Bean use of combinations of drugs containing an artemisinin Beetle, Onion Thrips, Parsleyworm, Pepper Maggot, Pepper derivative. Severe malaria is treated with intravenous or Weevil, Pickleworm, Potato Aphid, Potato Tuberworm, intramuscular quinine or, increasingly, the artemisinin Raspberry Crown Borer, Rednecked Cane Borer, Rhubarb 45 derivative artesunate. Several drugs are also available to Curculio, Root-knot Nematode, Rose Chafer, Rose Scale, prevent malaria in travellers to malaria-endemic countries Sap Beetles, Sawtoothed Grain Beetle, Wireworms, Squash (prophylaxis). Resistance has developed to several antima Bug, Squash Vine Borer, Tarnished Plant Bug, Twig Girdler/ larial drugs, most notably chloroquine. Twig Pruner, Vegetable Weevil, Virginia Pine, Sawfly, Malaria transmission can be reduced by preventing mos Wheel Bug, White Grubs, Whitefringed Beetles, Winter 50 quito bites by distribution of inexpensive mosquito nets and Grain Mite, and Yellow Ant. insect repellents, or by mosquito-control measures such as 2. Mosquito-Borne Disease spraying insecticides inside houses and draining standing Mosquitoes are a vector agent that carry disease-causing water where mosquitoes lay their eggs. viruses and parasites from person to person without catching Although many are under development, the challenge of the disease themselves. The principal mosquito borne dis 55 producing a widely available vaccine that provides a high eases are the viral diseases yellow fever, dengue fever and level of protection for a sustained period is still to be met. Chikungunya, transmitted mostly by the Aedes aegypti, and b. Helminthiasis malaria carried by the genus Anopheles. Though originally Some species of mosquito can carry the filariasis worm, a public health concern, HIV is now thought to be almost a parasite that causes a disfiguring condition (often referred impossible for mosquitoes to transmit. 60 to as elephantiasis) characterized by a great Swelling of Mosquitoes are estimated to transmit disease to more than several parts of the body; worldwide, around 40 million 700 million people annually in Africa, South America, people are living with a filariasis disability. The thread-like Central America, Mexico and much of Asia, with millions of filarial nematodes (roundworms) are members of the Super resulting deaths. At least 2 million people annually die of family Filarioidea, also known as “filariae. There are 9 these diseases. 65 known filarial nematodes which use humans as the definitive Methods used to prevent the spread of disease, or to host. These are divided into 3 groups according to the niche protect individuals in areas where disease is endemic include within the body that they occupy: lymphatic filariasis, sub US 9,578,881 B2 41 42 cutaneous filariasis, and serous cavity filariasis. Lymphatic epidemics caused by multiple serotypes (hyperendemicity) filariasis is caused by the worms Wuchereria bancrofti, can occur. Dengue is transmitted to humans by the Aedes Brugia malayi, and Brugia timori. These worms occupy the (Stegomyia) aegypti or more rarely the Aedes albopictus lymphatic system, including the lymph nodes, and in chronic mosquito. The mosquitoes that spread dengue usually bite at cases these worms lead to the disease elephantiasis. Subcu dusk and dawn but may bite at any time during the day, taneous filariasis is caused by loa loa (the African eye especially indoors, in shady areas, or when the weather is worm), Mansonella Streptocerca, Onchocerca volvulus, and cloudy. The WHO says some 2.5 billion people, two fifths of Dracunculus medimensis (the guinea worm). These worms the world's population, are now at risk from dengue and occupy the Subcutaneous layer of the skin, in the fat layer. estimates that there may be 50 million cases of dengue Serous cavity filariasis is caused by the worms Mansonella 10 perstans and Mansonella Ozzardi, which occupy the serous infection worldwide every year. The disease is now endemic cavity of the abdomen. In all cases, the transmitting vectors in more than 100 countries. are either blood Sucking insects (flies or mosquitoes), or Other viral diseases like epidemic polyarthritis, Rift Val copepod crustaceans in the case of Dracunculus medimensis. ley fever, Ross River Fever, St. Louis encephalitis, West Nile Individuals infected by filarial worms may be described as 15 virus (WNV), Japanese encephalitis, La Crosse encephalitis either “microfilaraemic' or “amicrofilaraemic.” depending and several other encephalitis type diseases are carried by on whether or not microfilaria can be found in their periph several different mosquitoes. Eastern equine encephalitis eral blood. Filariasis is diagnosed in microfilaraemic cases (EEE) and Western equine encephalitis (WEE) occurs in the primarily through direct observation of microfilaria in the United States where it causes disease in humans, horses, and peripheral blood. Occult filariasis is diagnosed in amicrofi some bird species. Because of the high mortality rate, EEE laraemic cases based on clinical observations and, in some and WEE are regarded as two of the most serious mosquito cases, by finding a circulating antigen in the blood. borne diseases in the United States. Symptoms range from c. Viruses mild flu-like illness to encephalitis, coma and death. Culex The viral disease yellow fever, an acute hemorrhagic and Culiseta are also involved in the transmission of disease. disease, is transmitted mostly by Aedes aegypti mosquitoes. 25 WNV has recently been a concern in the United States, The virus is a 40 to 50 nm enveloped RNA virus with prompting aggressive mosquito control programs. positive sense of the Flaviviridae family. The yellow fever d. Transmission virus is transmitted by the bite of female mosquitoes (the A mosquito's period of feeding is often undetected; the yellow fever mosquito, Aedes aegypti, and other species) bite only becomes apparent because of the immune reaction and is found in tropical and Subtropical areas in South 30 it provokes. When a mosquito bites a human, she injects America and Africa, but not in Asia. The only known hosts of the virus are primates and several species of mosquito. saliva and anti-coagulants. For any given individual, with The origin of the disease is most likely to be Africa, from the initial bite there is no reaction but with subsequent bites where it was introduced to South America through the slave the body's immune system develops antibodies and a bite trade in the 16th century. Since the 17th century, several 35 becomes inflamed and itchy within 24 hours. This is the major epidemics of the disease have been recorded in the usual reaction in young children. With more bites, the Americas, Africa and Europe. In the 19th century, yellow sensitivity of the human immune system increases, and an fever was deemed one of the most dangerous infectious itchy red hive appears in minutes where the immune diseases. response has broken capillary blood vessels and fluid has Clinically, yellow fever presents in most cases with fever, 40 collected under the skin. This type of reaction is common in nausea, and pain and it generally subsides after several days. older children and adults. Some adults can become desen In some patients, a toxic phase follows, in which liver sitized to mosquitoes and have little or no reaction to their damage with jaundice (giving the name of the disease) can bites, while others can become hyper-sensitive with bites occur and lead to death. Because of the increased bleeding causing blistering, bruising, and large inflammatory reac tendency (bleeding diathesis), yellow fever belongs to the 45 tions, a response known as Skeeter Syndrome. group of hemorrhagic fevers. The WHO estimates that 3. Insect Olfactory Receptors yellow fever causes 200,000 illnesses and 30,000 deaths The ability to detect and respond to the chemical envi every year in unvaccinated populations; around 90% of the ronment is a critical sensory input into many essential infections occur in Africa. behaviors of hematophagous (blood-feeding) insects A safe and effective vaccine against yellow fever has 50 (Zwiebel and Takken, 2004) (FIG. 1). The search for ver existed since the middle of the 20th century and some tebrate blood meals typically involves a flight of some countries require vaccinations for travelers. Since no therapy distance to reach the host. This behavior consists of a series is known, Vaccination programs are, along with measures to of behavioral stages, beginning with the activation of a reduce the population of the transmitting mosquito, of great receptive insect by the host chemical odor (kairomone) and importance in affected areas. Since the 1980s, the number of 55 ending when the insect alights on the host (Takken, 1991). cases of yellow fever has been increasing, making it a At close range, attraction is mediated by several odorants, reemerging disease. one of which is CO. In combination with other host-derived Dengue fever and dengue hemorrhagic fever (DHF) are organic chemicals, CO acts as a synergist as it greatly acute febrile diseases also transmitted by Aedes aegypti enhances the attraction triggered by other volatiles (Gilles, mosquitoes. These occur in the tropics, can be life-threat 60 1980). Moreover, it appears that mosquitoes respond to ening, and are caused by four closely related virus serotypes changes in the concentration of CO., rather than its presence of the genus Flavivirus, family Flaviviridae. It is also known or absence. In Ae. aegypti, changes in the firing rate of CO as breakbone fever, since it can be extremely painful. It receptors have been observed with increases in concentra occurs widely in the tropics, and increasingly in Southern tion of as little as 0.01% (Kellogg, 1970), while alterations China. Unlike malaria, dengue is just as prevalent in the 65 in behavior have been observed after increases of 0.03 urban districts of its range as in rural areas. Each serotype is 0.05% (Eiras and Jepson, 1991). Furthermore, a close sufficiently different that there is no cross-protection and examination of the role of CO revealed that the turbulence US 9,578,881 B2 43 44 of the odor plume in the laboratory greatly affected the IP3 increase is dependent on either a God or a Goto responsiveness of Ae. aegypti and An... gambiae S.S. (Dekker G-protein subunit. More recently, the inventors carried out a et al., 2001a). molecular survey of G-protein expression in the olfactory An... gambiae has also been shown to be attracted to appendages of An... gambiae, in which Gaq localization acetone, lactic acid (Acree et al., 1968), carboxylic acids consistent with involvement in olfactory signal transduction (Meijerink and van Loon, 1999), ammonia, 4-methyl-phe was observed along the dendrites of most olfactory sensory nol, 1-octen-3-ol, and other components of Sweat (Cork and neurons (Rutzler et al., 2006). Furthermore, pheromone Park, 1996; Meijerink et al., 2001), as well as to the odor of receptor neuron activity of Bombyx mori could be stimulated human feet, expired air and several unidentified components with fluoride ions (Laue et al., 1997), which are known to of Limburger cheese (De Jong and Knols, 1995). Further 10 activate heterotrimeric G proteins via binding to the a more, the often-cited differences in human attractiveness for Subunit in combination with magnesium ions (Antonny et mosquitoes (Curtis, 1986) is almost certainly olfactory al., 1993). However, despite this growing wealth of infor based (Qiu et al., 2006a; Schreck et al., 1990). This within mation, the precise mode of insect olfactory signal trans host differential behavior is most particularly expressed in duction remains largely obscure and is therefore the Subject anthropophilic culicids such as Ae. aegypti and An... gambiae 15 of ongoing investigation that has raised serious issues with s.s. (de Jong and Knols, 1995; Lindsay et al., 1993; Schreck regard to the validity of GPCR-based paradigms. et al., 1990). Host age, but not gender, may affect these Because olfaction was mediated by GPCRs in both ver inter-individual differences (Carnevale et al., 1978); race tebrates and at least one invertebrate, it was assumed that also appears to have no effect (Schreck et al., 1990). Young insects would also utilize these proteins in olfactory signal children have been shown to be less attractive to Anoph transduction. Indeed, using a variety of approaches, a large elines than adults (Muirhead-Thomson, 1951; Thomas, family of candidate ORS has been identified in D. melano 1951). Studies on the chemical composition of human gaster (Clyne et al., 1999) (Gao and Chess, 1999; Vosshall volatiles (Bernier et al., 1999; Krotoszynski et al., 1977: et al., 1999). In the first of these studies, putative D. Labows, 1979) revealed the existence of a large number melanogaster ORs (Dors) were identified using a novel (>350) of chemicals, and work is in progress to study the 25 computer algorithm that searched for conserved physico most important components of these volatiles regulating chemical features common to known transmembrane pro mosquito behavior. Lastly, it is also clear that responses to teins (Kim et al., 2000) rather than relying on a sequence CO affect inter-individual differences in attractiveness homology-based screen (which might miss a divergent (Brady et al., 1997) and, thus, CO serves as a universal member of a particular family). The structures that were attractant to many mosquito species (Gillies, 1980; Takken 30 ultimately identified using these strategies led to the iden et al., 1997: Takken and Knols, 1999). It has been reported tification of a highly divergent family of receptors, display that CO stimulation synergizes with host body odor and has ing between 10% and 75% identity and bearing no signifi an activating effect on host-seeking anopheline mosquitoes, cant homology to any other GPCR family (Smith, 1999). inducing take-off and sustained flight behaviors (Dekker et Another chemosensory receptor family was also described al., 2001b: Gillies, 1980; Mboera and Takken, 1997). 35 in D. melanogaster and An... gambiae and is presumed to In a process that is analogous to the sense of Smell in comprise gustatory (taste) receptors (Clyne et al., 2000: Hill humans as well as other insects, mosquito olfactionis initi et al., 2002; Scott et al., 2001). The other circumstantial ated by the process of chemosensory signal transduction by criterion to infer olfactory function has been provided by which chemical signals (typically environmental cues) are various in situ expression pattern studies that have demon translated into neuronal activity and, ultimately, behavioral 40 strated that the majority of these genes were selectively and outputs. In An... gambiae, this takes place within specialized Stereotypically expressed in the fly olfactory sensory neu hair-like structures called sensilla that are dispersed through rons (Clyne et al., 1999) (Elmore and Smith, 2001; Gao and out the antennae and other head appendages on adult and Chess, 1999; Vosshall, 2001; Vosshall et al., 1999). Two larval-stage anopheline mosquitoes (Zwiebel and Takken, color (double-labeling) in situ hybridization Suggests that, 2004) (FIG. 2). 45 with two notable caveats (Goldman et al., 2005), most D. Until recently, much of the inventors’ view of insect melanogaster ORNs are likely to express a single DOR gene olfactory signal transduction at the molecular level has been (Vosshall et al., 2000), which is analogous to mammalian strongly influenced by observations made in vertebrates, systems (Mombaerts, 1999), but in stark contrast to the C. crustaceans and nematodes (Hildebrand and Shepherd, elegans system. One apparent exception to the one ORN 1997: Krieger and Breer, 1999). The canonical model 50 one receptor principle is the non-conventional DORco. involves a family of heptahelical G-protein-coupled recep Unlike most other DORs, DORco is expressed throughout tors (GPCRs) that activate downstream effectors via het the majority of antennal and maxillary palp ORNs of D. erotrimeric GTP-binding (G) proteins and traditional second melanogaster. Putative DORco orthologs have been identi messengers. It has long been assumed, although not fully fied in a wide range of insect species and share many accepted (see below), that the canonical model of olfactory 55 characteristics, including high sequence identity (Pitts et al., signal transduction would also hold true in insects, in which 2004), characteristic broad expression pattern (Krieger et al., several of the “usual molecular suspects have been identi 2003) and conserved functions (Jones et al., 2005). DORco fied and, in part, functionally characterized. These include family members are considered non-conventional ORS as arrestins (Merrill et al., 2002; 2003; 2005), odorant-binding they act as general dimerization partners for other members proteins (OBPs) (Pelosi and Maida, 1995), a heterotrimeric 60 of the DOR family (Larsson et al., 2004). More recently, G-protein (Laue et al., 1997) as well as a CNG (Baumann et Benton, Vosshall and co-workers have identified a novel set al., 1994: Krieger et al., 1999) and an IP3-gated ion channel of ionotropic glutamate receptors as a new class of insect (Stengl. 1994). In one study using the cockroach, it was chemosensory receptors (IRs) that are expressed in DOr83 demonstrated that pheromone exposure of insect antennal ORNs associated with coeloconic sensilla where they act in preparations caused a rapid increase in IP3 levels (Breer et 65 parallel with “classical insect ORs to respond to ammonia al., 1990), which in a follow-up study could be inhibited by and other environmental cues (Benton et al., 2009; Liu et al., pertussis toxin (Boekhoff et al., 1990), indicating that the 2010). US 9,578,881 B2 45 46 Elegant studies by the Vosshall lab have also suggested a set of four odorants. The Carlson laboratory has used a that insect ORS manifest a novel topology relative to ver novel experimental approach that takes advantage of a tebrate ORs (Benton et al., 2006). In the absence of actual genetic strain of D. melanogaster in which a chromosomal structural information insect ORS have been structurally deletion has resulted in the loss of the endogenous receptors characterized largely based on bioinformatic models derived (DOR2a/b) from the ab3AORN. The resultant formation of from vertebrates (Clyne et al., 2000; Vosshall et al., 1999). a “empty neuron' system facilitates the specific targeting of Indeed, while sequence-based phylogenies recognize that exogenous OR genes into the empty neuron, thereby allow insect ORS in general comprise a distinct family of hepta ing electrophysiological assessment of the ability of the helical receptors that are an expanded lineage of ancestral novel receptor to carry out chemosensory signal transduc tion within the ab3A neuron upon stimulation with a diverse chemosensory receptors (Mombaerts, 1999; Robertson et 10 set of odorants (Dobritsa et al., 2003). This system has been al., 2003) there is a growing awareness that insect ORs are used effectively to functionally characterize nearly all the likely to represent a structurally unique set of sensory DORs (Hallem et al., 2004a) (Hallem and Carlson, 2006), proteins. These studies provide compelling evidence in leading to a highly developed map of the multidimensional support of the view that Drosophila ORs are heteromeric “odor space of the DORs. As part of a long-standing complexes between the non-conventional DOR83b and con 15 collaboration between the Carlson lab and that of the inven ventional, odorant binding DORs that adopt a novel mem tors, multiple AgORs have also been functionally charac brane topology in which the N-terminus is intracellular terized in the Drosophila empty neuron (Hallem et al., rather than the extra-cellular localization that is typical of 2004b; Lu et al., 2007). These studies, along with the vertebrate ORs and GPCRs (Benton et al., 2006). Indepen Success in functionally expressing over 40 AgORS in Xeno dent validation (Lundin et al.) together with recent compu 20 pus and cell culture systems, have lead to significant tational analyses employing hidden Markov modeling that advances in understanding the molecular basis for olfactory “strongly rejects' classifying arthropod ORs as GPCRs sensitivity in larval (Xia et al., 2008) and adult (Lu et al., (Wistrand et al., 2006) raise significant concerns regarding 2007) An... gambiae. For example, CO, which acts as uni the nature of the signaling pathways that are downstream of versal attractant for many species of mosquitoes (Takken odorant activation in insects. Indeed, two recent studies 25 and Knols, 1999), elicits avoidance in Drosophila where it provide provocative evidence to Suggest that Drosophila has been identified as an active component of the “stress ORs manifest properties of both ligand-gated (Sato et al.) odorant’ that targets a discrete population of sensory neu rons (Suh et al., 2007) and where a pair of highly conserved and cyclic-nucleotide-gated ion channels (Wicher et al., putative gustatory receptors {Gr21a and Gró3a) have been 2008). While these hypotheses still differ in their particulars, shown to both be both necessary and sufficient to mediate there is growing awareness that insect olfactory transduction 30 olfactory sensitivity to CO in Drosophila (Jones et al., may diverge from vertebrate paradigms and act as non 2007; Kwon et al., 2007). As part of a comprehensive study GPCR-mediated ion-channels (FIG. 2). In any case, while of the olfactory processes on the maxillary palp in An current hypotheses may differ, the growing possibility that gambiae, the inventors have identified three Gr21a/63a insect olfactory transduction may diverge from vertebrate homologs (AgCrs22-24) as the molecular partners required paradigms and act via non-GPCR-mediated mechanisms 35 that together comprise the anopheline CO2 receptor (Lu et Such as ion channels (FIG. 2) is compelling. al., 2007). In the first report of insect ORS outside of the model insect C. Compounds system D. melanogaster, members of the inventors’ labo In one aspect, the invention relates to a compound having ratory, as part of a collaborative effort with Drs. John a structure represented by a formula: Carlson and Hugh Robertson, were responsible for the 40 identification of a set of candidate Or genes selectively expressed in olfactory tissues of An... gambiae (AgORS) (Fox et al., 2001). Moreover, that report also demonstrated that at N1\ least one of the initial set of AgORs displays female-specific expression, a feature that may be especially relevant for 45 disease transmission. In a Subsequent study, as part of the effort to annotate the recently completed genomic sequence l! ) of An... gambiae (Holt et al., 2002), the inventors (in col laboration with other groups) utilized bioinformatics and molecular approaches to describe the entire An... gambiae 50 GPCR gene family (AgCPCRs); of the 275 putative AgG wherein: R is hydrogen or is taken together with R to be PCRs, 79 candidate AgORs were described (Hill et al., alkanediylcar alkenediylcar or a substituted version of 2002). Furthermore, a similar bioinformatic approach (using either of these groups; R is hydrogen or is taken together a non-public database) has been used to identify nine with R" as defined above; and R is hydrogen, hydroxy, candidate Or genes in the heliothine moth Heliothis vire 55 nitro, halo, alkylos Substituted alkylcs, alkenyls, or scens (Krieger et al., 2002), Some of which share sequence substituted alkenyls, or a salt or tautomer of the formula. homology with AgORs. More recently, a large family of In various aspects, the invention relates to a compound candidate Or genes have been identified in the genome having a structure represented by a formula: sequence of the honey bee, Apis mellifera (Robertson and Wanner, 2006), Ae. aegypti (Bohbot et al., 2007) and the red 60 N-N flour beetle, Tribolium casteneum (EngSontia et al., 2008). Thus far, insect ORS have been extensively deorphanized X-s O in a number of heterologous systems. The first Successful functional studies of insect ORs were carried out for N RI DOR, using a Xenopus oocyte expression system (Wetzel 65 C. : et al., 2001), and over-expression in D. melanogaster (Storkuhl and Kettler, 2001) showed increased sensitivity to US 9,578,881 B2 47 48 wherein: R' is hydrogen or is taken together with R to be wherein R' is -H, -OH, -F, - C1, —Br. —I, NH, alkanediyl ca, alkenediyl or a substituted version of —NO, COH, -COCH, —CN, -SH, —OCH, either of these groups; R is hydrogen or is taken together –OCHCH, C(O)CH, N(CH), C(O)NH - OC with R" as defined above; and R is hydrogen, hydroxy, (O)CH, or - S(O)NH2. nitro, halo, alkylos. Substituted alkylos), alkenyls, or 5 substituted alkenyls, or a salt or tautomer of the formula, In a further aspect, the compound is further defined by the provided that the compound is not: formula: Y-C- 10 C’s O Orcu 15 "t - N / In a further aspect, the invention relates to a compound of RI the formula: R2

In a further aspect, the compound is further defined by the O formula: 4 R R6 s / \ -R, 25 Z r -- wherein: R is hydrogen or is taken together with R to be 30 N 2 2 \- alkanediylca, alkenediyl or a substituted version of N either of these groups; R2 is hydrogen, alkyls substituted / alkyls, or is taken together with R as defined above: R. R R3. is hydrogen, hydroxy, nitro, halo, alkyls substituted R2 alkylcs, alkenyles, or Substituted alkenyles: R is alkylc-s) alkenylcs arylc-10): aralkylcio 35 heteroaryles, heteroaralkyls, or substituted versions of In a further aspect, the compound is further defined by the any of these groups; Rs is heteroaryl or substituted formula: heteroaryl; and R is hydrogen, alkyls. Substituted alkylos, alkenyls, or Substituted alkenyls, or a salt or tautomer of the formula; provided that if R and Rare H 40 and Rs is 3-pyridinyl, then Rs cannot be ethyl. In a further aspect, the compound is further defined by the formula: r (-)- N 2 45

O R4

N 50 R R3. wherein: R is hydrogen or is taken together with R to be In a further aspect, the compound is further defined by the alkanediylcar alkenediylcar or a substituted version of formula: 55 either of these groups; R2 is hydrogen, alkyls, substituted alkyls, or is taken together with R as defined above. Rs is hydrogen, hydroxy, nitro, halo, alkyls substituted alkylos), alkenylcs), or Substituted alkenyls). R4 is R'll Y -- 60 alkylc-s): alkenylcs). arylc-10): aralkylcio. heteroarylcs, heteroaralkylcs, or substituted versions of (C I - any of these groups; and R' is -H, -OH, - F. —Cl, Br, N I, NH, NO. —COH, -COCH, —CN, -SH, R1 65 —OCH, OCHCH. —C(O)CH, N(CH), —C(O) NH, —OC(O)CH, or—S(O)NH2 or a salt or tautomer of the formula, wherein the compound is not: US 9,578,881 B2 49

In a further aspect, the compound is further defined by the formula:

In various aspects, the invention relates to a compound Xa ly, O having a structure represented by a formula: o,

In various aspects, the invention relates to a compound 25 wherein m, n, p, and q are independently 0 or 1; wherein L' having a structure represented by a formula: and L are independently divalent organic groups having from 1 to 8 non-hydrogen members; wherein Q' is —O , S-, -S(O)-, or -S(O) ; wherein Q is —O , —S , or NR'; wherein R is optionally substituted and 30 selected from monocyclic aryl, bicyclic aryl, monocyclic heteroaryl, bicyclic heteroaryl, and tricyclic heteroaryl; wherein R' is hydrogen, optionally substituted C1-C4 alkyl, optionally substituted phenyl, optionally substituted benzyl, or a structure represented by a formula selected from: 35 wherein m, n, p, and q are independently 0 or 1; wherein L' O O O and L are independently divalent organic groups having from 1 to 8 non-hydrogen members; wherein Q' is —O , A-s-s A-si A-. S-, -S(O)—, or -S(O) ; wherein Q is —O , 40 —S , or NR'; wherein R is optionally substituted and selected from monocyclic aryl, bicyclic aryl, monocyclic or R' is taken together with a substituent of R to form a five-, six-, or seven-membered heterocylcoalkyl ring; heteroaryl, bicyclic heteroaryl, and tricyclic heteroaryl; wherein R is optionally substituted and selected from (C1 wherein R' is hydrogen, optionally substituted C1-C4 alkyl, C5) alkyl, (C1-C5) alkenyl, (C6-C10) aryl, (sC10) aralkyl, optionally substituted phenyl, optionally substituted benzyl, 45 ( a- A-. or a salt or tautomer thereof, wherein the compound is not: or R' is taken together with a substituent of R to form a five-, six-, or seven-membered heterocylcoalkyl ring; 55 wherein R is optionally substituted and selected from (C1 C5) alkyl, (C1-C5) alkenyl, (C6-C10) aryl, (sC10) aralkyl, \- ) -- (sC8) heteroaryl, and (sC8) heteroaralkyl; wherein R is le s O H s optionally substituted aryl or optionally substituted (sC6) 60 Urrrol heteroaryl; and wherein R* and R' are independently selected from hydrogen, optionally substituted (C1-C5) alkyl, or optionally substituted (C1-C5) alkenyl, or R'' and R', along with the intermediate carbon, together comprise 65 a C3-C6 cycloalkyl ring or a C2-C5 heterocylcoalkyl ring: or a salt or tautomer thereof, wherein the compound is not: US 9,578,881 B2 51 52 or wherein the compound is not: In a further aspect, the compound comprises:

N N N R4 X S O RetyCa N -s O N N N \ { R85 \ y- x- | -R3 % > N R3, N-N 6, R65 le s R1 RI 10 R2 R2 wherein R is optionally substituted and selected from (C1 wherein: R is hydrogen or is taken together with R to be C5) alkyl, (C1-C5) alkenyl, (C6-C10) aryl, (sC10) aralkyl, alkanediylca, alkenediylca, or a substituted version of 15 (sC8) heteroaryl, and (sC8) heteroaralkyl. either of these groups; R is hydrogen or is taken together In a further aspect, wherein the compound is not: with R" as defined above; and R is hydrogen, hydroxy, nitro, halo, alkylos. Substituted alkylos), alkenylcs, or substituted alkenyls, or a salt or tautomer of the formula. In a further aspect, the compound comprises: ---- R4 Na2

25 "Scy-\ | O ( N) N-N R6a R6b le s RI R2 30 wherein: R is hydrogen or is taken together with R to be alkanediylcar alkenediylcar or a substituted version of wherein R is hydrogen, hydroxy, nitro, halo, optionally either of these groups; R2 is hydrogen, alkyls. Substituted substituted (C1-C5) alkyl, optionally substituted (C2-C5) alkyls, or is taken together with R as defined above. Rs alkenyl, or optionally substituted (C2-C5) alkynyl; or R is is hydrogen, hydroxy, nitro, halo, alkyls substituted taken together with R' to be optionally substituted (C1-C4) 35 alkylos), alkenylcs), or Substituted alkenyls). R4 is alkanediyl or optionally substituted (C1-C4) alkenediyl; and alkylc-s): alkenylcs). arylc-10): aralkylcio. wherein R is hydrogen, hydroxy, nitro, halo, optionally heteroarylcs, heteroaralkylcs, or substituted versions of substituted (C1-C5) alkyl, optionally substituted (C2-C5) any of these groups; and R'' is -H, -OH, - F. —Cl, Br, alkenyl, or optionally substituted (C2-C5) alkynyl. I, NH, NO. —COH, -COCH —CN. —SH, 40 —OCH, OCHCH. —C(O)CH, N(CH), —C(O) In a further aspect, the compound comprises: NH, —OC(O)CH, or - S(O)NH, or a salt or tautomer of the formula. In a further aspect, the compound comprises: N 45 Q O N y-4 Q N N \ y- x- | -R3 Ra-H- N-N R6a R6b le s y 21 S-Q) O N R1 50 R2 R85 \ y- x- | -R3 N-N 6, R6b led wherein R is hydrogen, hydroxy, nitro, halo, optionally RI R2 substituted (C1-C5) alkyl, optionally substituted (C2-C5) alkenyl, or optionally substituted (C2-C5) alkynyl; or R is 55 wherein R is hydrogen, hydroxy, nitro, halo, optionally taken together with R' to be optionally substituted (C1-C4) substituted (C1-C5) alkyl, optionally substituted (C2-C5) alkanediyl or optionally substituted (C1-C4) alkenediyl: alkenyl, or optionally substituted (C2-C5) alkynyl; or R is wherein R is hydrogen, hydroxy, nitro, halo, optionally taken together with R' to be optionally substituted (C1-C4) substituted (C1-05) alkyl, optionally substituted (C2-C5) alkanediyl or optionally substituted (C1-C4) alkenediyl: alkenyl, or optionally substituted (C2-C5) alkynyl; and 60 wherein R is hydrogen, hydroxy, nitro, halo, optionally wherein R* and Rare independently selected from hydro substituted (C1-C5) alkyl, optionally substituted (C2-C5) gen, hydroxy, nitro, halo, optionally substituted (C1-C5) alkenyl, or optionally substituted (C2-C5) alkynyl; and alkyl, or optionally substituted (C1-C5) alkenyl; or R and wherein R* and Rare independently selected from hydro R” are positioned on adjacent carbons and are taken 65 gen, hydroxy, nitro, halo, optionally substituted (C1-C5) together to be optionally substituted (C1-C4) alkanediyl or alkyl, or optionally substituted (C1-C5) alkenyl: or R and optionally substituted (C1-C4) alkenediyl. R" are positioned on adjacent carbons and are taken US 9,578,881 B2 53 54 together to be optionally substituted (C1-C4) alkanediyl or wherein R is hydrogen, hydroxy, nitro, halo, optionally optionally substituted (C1-C4) alkenediyl. substituted (C1-C5) alkyl, optionally substituted (C2-C5) In a further aspect, the compound comprises: alkenyl, or optionally substituted (C2-C5) alkynyl: or R is taken together with R' to be optionally substituted (C1-C4) alkanediyl or optionally substituted (C1-C4) alkenediyl: N1N wherein R is hydrogen, hydroxy, nitro, halo, optionally Ra-H- substituted (C1-C5) alkyl, optionally substituted (C2-C5) '4N-N-s, if alkenyl, or optionally substituted (C2-C5) alkynyl; and N wherein R* and Rare independently selected from hydro 10 *1)(x-OY,N-N R6a R6b e s gen, hydroxy, nitro, halo, optionally substituted (C1-C5) R1 alkyl, or optionally substituted (C1-C5) alkenyl: or R and R2 R" are positioned on adjacent carbons and are taken together to be optionally substituted (C1-C4) alkanediyl or wherein R is optionally substituted and selected from (C1 15 optionally substituted (C1-C4) alkenediyl. C5) alkyl, (C1-C5) alkenyl, (C6-C10) aryl, (sC10) aralkyl, In a further aspect, the compound comprises: (sC8) heteroaryl, and (sC8) heteroaralkyl. In a further aspect, the compound comprises:

N R4 R8 - 1s O y 21 Q Q O N y- N R85 \ | ( ). 25 N-N R6a R6b le s \N-N - R6ax- R6b | le -R3 s R1 RI R2 R2 wherein R is hydrogen, hydroxy, nitro, halo, optionally substituted (C1-C5) alkyl, optionally substituted (C2-C5) 30 alkenyl, or optionally substituted (C2-C5) alkynyl; or R is wherein R is optionally substituted and selected from (C1 taken together with R' to be optionally substituted (C1-C4) C5) alkyl, (C1-C5) alkenyl, (C6-C10) aryl, (sC10) aralkyl, alkanediyl or optionally substituted (C1-C4) alkenediyl: (sC8) heteroaryl, and (sC8) heteroaralkyl. wherein R is hydrogen, hydroxy, nitro, halo, optionally In a further aspect, the compound comprises: substituted (C1-C5) alkyl, optionally substituted (C2-C5) 35 alkenyl, or optionally substituted (C2-C5) alkynyl; and wherein R* and Rare independently selected from hydro gen, hydroxy, nitro, halo, optionally substituted (C1-C5) allyl, or optionally substituted (C1-C5) alkenyl; or R and R” are positioned on adjacent carbons and are taken 40 ( \ . O together to be optionally substituted (C1-C4) alkanediyl or 3 Q N optionally substituted (C1-C4) alkenediyl. In a further aspect, the compound comprises: N-N R6a R6b le s N R1 45 R2 N O O21 S N *1)(x-OY,N-N R6a R6b le s wherein Q is —O , —S , or NR; wherein R is R1 R2 hydrogen, hydroxy, nitro, halo, optionally Substituted (C1 C5) alkyl, optionally substituted (C2-C5) alkenyl, or option wherein R is optionally substituted and selected from (C1 ally substituted (C2-C5) alkynyl: or R is taken together with C5) alkyl, (C1-C5) alkenyl, (C6-C10) aryl, (sC10) aralkyl, R" to be optionally substituted (C1-C4) alkanediyl or option (sC8) heteroaryl, and (sC8) heteroaralkyl. 55 In a further aspect, the compound comprises: ally substituted (C1-C4) alkenediyl; wherein R is hydrogen, hydroxy, nitro, halo, optionally substituted (C1-C5) alkyl, optionally substituted (C2-C5) alkenyl, or optionally sub

60 stituted (C2-C5) alkynyl; wherein R is optionally substi 1s tuted and selected from (C1-C5) alkyl, (C1-C5) alkenyl, y 2 Q Q O N (C6-C10) aryl, (sC10) aralkyl, (sC8) heteroaryl, and (sC8) R85 \ - x- | -R3 heteroaralkyl; and wherein each of R', R', and R' is N-N 6, R6b le s independently selected from hydrogen, hydroxy, nitro, halo, R1 R2 optionally substituted (C1-C5) alkyl, or optionally substi tuted (C1-C5) alkenyl: or any two of R', R', and R'' are US 9,578,881 B2 55 56 positioned on adjacent carbons and are taken together to be In a further aspect, the compound binds to and/or modu optionally substituted (C1-C4) alkanediyl or optionally sub lates insect Orco ion channels. stituted (C1-C4) alkenediyl. It is contemplated that each disclosed derivative can be In a further aspect, the compound comprises: optionally further substituted. It is also contemplated that any one or more derivative can be optionally omitted from the invention. It is understood that a disclosed compound can be provided by the disclosed methods. It is also under stood that the disclosed compounds can be employed in the disclosed methods of using. 10 1. Structure Suitable substituents are described below. R6a R65 le s a. L' Groups In one aspect, L' is a divalent organic groups having from 15 1 to 8 non-hydrogen members. For example, L' can have 1, 2, 3, 4, 5, 6, 7, or 8 non-hydrogen members. In a further wherein R is optionally substituted and selected from (C1 aspect, L' is selected from C5) alkyl, (C1-C5) alkenyl, (C6-C10) aryl, (sC10) aralkyl, (sC8) heteroaryl, and (sC8) heteroaralkyl. In a further aspect, the compound comprises: Ary /-X N 25 Q le Q O N \ y- Q x- | -R3 ^ry Ax /-y N-N R6a R6b le s R1 R2 In a further aspect, L' is present when p is 1. In a further aspect, L' is absent when p is 0. wherein Q is - O -, -S , or NR; wherein R is b. L. Groups hydrogen, hydroxy, nitro, halo, optionally Substituted (C1 In one aspect, L is a divalent organic groups having from C5) alkyl, optionally substituted (C2-C5) alkenyl, or option 35 1 to 8 non-hydrogen members. For example, Li can have 1, ally substituted (C2-C5) alkynyl; or R is taken together with 2, 3, 4, 5, 6, 7, or 8 non-hydrogen members. In a further R" to be optionally substituted (C1-C4) alkanediyl or option aspect, L is selected from ally substituted (C1-C4) alkenediyl; wherein R is hydrogen, hydroxy, nitro, halo, optionally substituted (C1-C5) alkyl, optionally substituted (C2-C5) alkenyl, or optionally sub 40 stituted (C2-C5) alkynyl; wherein R is optionally substi tuted and selected from (C1-C5) alkyl, (C1-C5) alkenyl, A-A a y A-X (C6-C10) aryl, (sC10) aralkyl, (sC8) heteroaryl, and (sC8) heteroaralkyl; and wherein each of R', R', and R' is independently selected from hydrogen, hydroxy, nitro, halo, 45 optionally substituted (C1-C5) alkyl, or optionally substi tuted (C1-C5)alkenyl: or any two of R', R', and R'' are positioned on adjacent carbons and are taken together to be optionally substituted (C1-C4) alkanediyl or optionally sub 50 /o, CA stituted (C1-C4) alkenediyl. In a further aspect, the compound comprises: In a further aspect, L is present when q is 1. In a further aspect, L is absent when q is 0. c. Q' Groups R10a 55 In one aspect, Q' is -O-, -S , —S(O)—, or S R10b f —S(O), . In a further aspect, Q" is —O— or -S-. In a Q further aspect, Q' is —O-. In a further aspect, Q' is —S-. le N S O In a further aspect, Q" is S(O) . In a further aspect, Q' y- | N is —S(O) . R10c \ f N -R3 60 d. Q' Groups N-N R6a R6b le s In one aspect, Q is —O —S , or NR. In a further R1 aspect, Q is —O-. In a further aspect, Q is —S-. In a R2 further aspect, Q is NR". e. Q. Groups wherein R is optionally substituted and selected from (C1 65 In one aspect, Q is —O , —S , or NR. In a further C5) alkyl, (C1-C5) alkenyl, (C6-C10) aryl, (sC10) aralkyl, aspect, Q is —O—. In a further aspect, Q is —S-. In a (sC8) heteroaryl, and (sC8) heteroaralkyl. further aspect, Q is —NR. US 9,578,881 B2 57 58 f. R' Groups further aspect, R4 is alkylos, alkenylcs), arylaco): In one aspect, R' is hydrogen, optionally substituted aralkylco, heteroaryles, heteroaralkylcs, or substi C1-C4 alkyl, optionally substituted phenyl, optionally sub tuted versions of any of these groups. In a further aspect, R. stituted benzyl, or a structure represented by a formula is alkyls for example, ethyl, propyl, or cyclopropyl. In 5 a further aspect, R is alkenyls for example, allyl. In a selected from: further aspect, R comprises a structure represented by a formula selected from: O O A-s-s a-s, and 10 A-.O 15 In a further aspect, R' is hydrogen. In a further aspect, R' is methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s-butyl, t-butyl, optionally substituted phe nyl, optionally substituted benzyl, or a structure represented by a formula selected from:

O O A-s-s a-s, and 25 O

A-. 30 In a further aspect, R' is hydrogen, and wherein R is hydrogen, hydroxy, nitro, halo, optionally Substituted (C1 C5) alkyl, optionally substituted (C2-C5) alkenyl, or option 35 ally substituted (C2-C5) alkynyl. In a further aspect, R' is taken together with a substituent of R to form a five-, six-, or seven-membered heterocyl coalkyl ring. In a further aspect, R' is taken together with a substituent of R7 to be optionally substituted (C1-C4) 40 alkanediyl or optionally substituted (C1-C4) alkenediyl. In a further aspect, R' is hydrogen or is taken together with R to be alkanediylcar alkenediylcula, or a substituted version of either of these groups. In a further aspect, R and R2 are taken together to be ethanediyl. 45 g. R Groups In one aspect, R is hydrogen, hydroxy, nitro, halo, optionally substituted (C1-C5) alkyl, or optionally substi tuted (C1-C5) alkenyl. In a further aspect, R is hydrogen. In a further aspect, R is hydrogen or is taken together with R' 50 as defined above. h. R. Groups In one aspect, R is hydrogen, hydroxy, nitro, halo, optionally substituted (C1-C5) alkyl, or optionally substi j. R Groups tuted (C1-C5) alkenyl. In a further aspect, R is hydrogen. In 55 In one aspect, R is optionally substituted aryl or option a further aspect, R is halo, for example, fluoro, chloro, or ally substituted (sC6) heteroaryl. In a further aspect, R is bromo. In a further aspect, R is alkyls. In a further pyridinyl, for example, 3-pyridinyl or 4-pyridinyl. In a aspect, the Rs alkylcs) has no quaternary carbon atoms. In further aspect, Rs is pyrazolyl or methylpyrazolyl. a further aspect, R is methyl, ethyl, n-propyl, or isopropyl. In a further aspect, R is selected from: In a further aspect, R is alkenyls. In a further aspect, Rs 60 is vinyl. In a further aspect, R is hydrogen, hydroxy, nitro, halo, alkylos). Substituted alkylos), alkenylcs, or Sub N stituted alkenyls. i. R Groups In one aspect, R is optionally substituted and selected 65 from (C1-C5) alkyl, (C1-C5) alkenyl (C6-C10) aryl, (sC10) aralkyl, (sC8) heteroaryl, and (sC8) heteroaralkyl. In a US 9,578,881 B2 59 60 -continued -continued N rt NN ) s A4 s 5 N R85 In a further aspect, R is selected from:

N R10c 10 N , and R10a Q3 s F 2 MeO

15 N N1 N Oy 2O 21 2 1 NS C/ 2 21 O O \/ MeO N 1n 1 C?y Na21. n 30 H V F N N1 N 35 N Oy and Na2

40 V

y 45 HN 5 /N 50 N N N In a further aspect, R is substituted with 0-3 groups independently selected from hydroxy, nitro, halo, carboxyl, O O ss carboxy(C1-C4)alkyl, phenyl, benzyl, benzyloxy, amino, e alkyl (C1-C4)amino, dialkyl (C1-C4, C1-C4)amino, C1-C4 N alkyoxyl, C1-C5 alkyl, and C1-C5 alkenyl. (S^ NNo. k. R Groups In one aspect, R'' and R' are independently selected Na l 60 from hydrogen, optionally substituted (C1-C5) alkyl, or / HN / optionally substituted (C1-C5) alkenyl. In a further aspect, R" is hydrogen. In a further aspect, R' is hydrogen. In a N further aspect, R is non-hydrogen. In a further aspect, R' N N and is non-hydrogen. In a further aspect, R" is optionally NNN N 65 substituted (C1-C5) alkyl or optionally substituted (C1-C5) H alkenyl. In a further aspect, R is optionally substituted (C1-C5) alkyl or optionally substituted (C1-C5) alkenyl. US 9,578,881 B2 61 62 In a further aspect, R' is methyl, ethyl, n-propyl, i-pro -continued pyl. n-butyl, i-butyl, s-butyl, or t-butyl. In a further aspect, R is methyl, ethyl, n-propyl, i-propyl. n-butyl, i-butyl, ; and O s-butyl, or t-butyl. In a further aspect, R'' and R', along with the intermediate carbon, together comprise a C3-C6 5 cycloalkyl ring or a C2-C5 heterocylcoalkyl ring. A ses2. /C)O 1. R. Groups / In one aspect, R7 is optionally substituted and selected from monocyclic aryl, bicyclic aryl, monocyclic heteroaryl, 10 bicyclic heteroaryl, and tricyclic heteroaryl. In a further In a further aspect, aspect, R7 is substituted. In a further aspect, R7 is unsubsti tuted. In a further aspect, R is monocyclic aryl or mono cyclic heteroaryl. In a further aspect, R is bicyclic aryl, bicyclic heteroaryl, or tricyclic heteroaryl. In a further 15 aspect, R is monocyclic aryl or bicyclic aryl. In a further aspect, R is monocyclic heteroaryl, bicyclic heteroaryl, or tricyclic heteroaryl. In a further aspect, R is substituted with 0-3 groups comprises a structure represented by a formula selected independently selected from hydroxy, nitro, halo, carboxyl, carboxy(C1-C4)alkyl, phenyl, benzyl, benzyloxy, amino, alkyl (C1-C4)amino, dialkyl (C1-C4, C1-C4)amino, C1-C4 alkyoxyl, C1C5 alkyl, C1-C5 alkenyl, and C1-C6 sulfona mido. 25 In a further aspect, R' comprises a structure represented by a formula selected from: N A C 30 ACH /

N / C / C / 35 40 / H C / C.

45 A.H 1CCOCH3 /O.H

Or N COCH 50 A. A. O. H H

55 A.ON AC H 49/5 60 O A.ON A. h N'- 65 /c/c, O US 9,578,881 B2 63 64 -continued -continued CF O

S H r A N A N OO h Br F A O A A. A. O N h 9N-1 H C 15 O OMe

H O H OMe 25

COCH3

30 N N O

H H C 35 A. H 40 O O 21

YusN

45 A N H h F 50 A. O N H H and Br

55

H H 9 m. R Groups OMe In one aspect, wherein each of R and R is indepen dently selected from hydrogen, hydroxy, nitro, halo, option ally substituted (C1-C5) alkyl, or optionally substituted N N NO (C1-C5) alkenyl. In a further aspect, R and R are posi 65 tioned on adjacent carbons and are taken together to be H H optionally substituted (C1-C4) alkanediyl or optionally sub stituted (C1-C4) alkenediyl. US 9,578,881 B2 65 66 n. R Groups -continued In one aspect, R is optionally substituted and selected from (C1-C5) alkyl, (C1-C5) alkenyl (C6-C10) aryl, (sC10) aralkyl, (sC8) heteroaryl, and (sC8) heteroaralkyl. o. R' Groups In one aspect, each of R', R', and R' is indepen dently selected from hydrogen, hydroxy, nitro, halo, option ally substituted (C1-C5) alkyl, or optionally substituted (C1-C5)alkenyl. In a further aspect, any two of R', R', and R'' are positioned on adjacent carbons and are taken together to be optionally substituted (C1-C4) alkanediyl or 10 optionally substituted (C1-C4) alkenediyl. p. R' Groups In one aspect, R' is —H, -OH, - F - C1, —Br. —I. NH, NO, COH, COCH, CN, -SH, —OCH, OCHCH. —C(O)CH, N(CH), —C(O) 15 NH, —OC(O)CH, or—S(O)NH2. In a further aspect, R, is hydrogen. In a further aspect, R, is fluoro. 2. Example Compounds In various aspects, the disclosed compounds can be pres ent having one or more structures represented by formulae listed below: In one aspect, a disclosed compound can have the formula (I):

25 (I)

Q- O 30 R6a R6b le worx C.2 Examplary compound within Formula (I) include, but are 35 not limited to: C - O 40 C (-)-O 45 / \

55

60

65 US 9,578,881 B2 67 68 ontinued / \ S O CY \ cro-ON -s O 5 -(r)-O 2 \ S O 10 S. Cr-C CY. cro-ON -s if 15 / \ N S O S. Cr-C Ctrl-O 25

Ctrl-O 30 s/{\- -O 35 es C cro- O -k - 40 ey C - O 45 -(r)- esO O try to 50 -k / \ S O C-cr-O 55 -(r)-O N N O h 60 / \ S S/h ( ) Ctrl-Of 65 US 9,578,881 B2 69 70 Y S/ ( ) - C C cro-O uo. CrxN r '-OO US 9,578,881 B2 71 72 s ( r^-J - (-\- O

O -O 15 O-(r^- S O O O --Or- O orO Ctrl-O\ N O Y \ N-N ON-R C2. s - O O AO -O 50 C -- O City-Os r O- O O 60- C/ y - O O US 9,578,881 B2 73 74 -continued -continued s No O --Or-O -(r^-O 10 o1 H

Ns N o1 C - O - -C -O

25 S. \ { N S. r N-O 30 C / O Ctrl-C2 O O C- (--O

S. (r^-O 50 2 \ C S O O\1 H S/ \N-N x- N -O Y T O 55 S Cr-OOr six-O - (r^- O - (r^- O US 9,578,881 B2 75 76 -continued -continued O O cru, O I C - O - -SCr Cut 15 H3CO N O'CN syCr-O C -u -O C rulo (r^-O

CutN-N N 35 -N4 / \ O'CN 7CN- \ )N ru,O -O " S.- (--O\ { NH \ H N O Y N O S. Cry--O O---, -C - N Z \ N O /CO Z \ -- -O FC S. {r^-OH f N N 55 O is/ \ , O ( ) -Nils i f S. {r^-O FC S. S. N-O US 9,578,881 B2 77 78

N N O

\ N O H

C.,S. (r^- O O 15 /s/Y \ h r^-,O -O HS Ctrl-C/ \ r O - d.N y , -Or-,-O - K N r 30 N-N S S. S. N-O W r O

/A-fu's O ) S O --Or-ON-N H \N N- -O a r O

YN \ Nr O sh,N-N N-K N -O S. (r^-O 45 O r

US 9,578,881 B2 81 82 -continued -continued CS/h (Cre, ) O O- C--'N l(-N-O-4 s if N- ) F-NaO---, \ N O ON 15 CN -u O-4

N-N N S/h (SN-l \-s N-N N N 7 \ O

S/Sky-Null 65 N-N N

US 9,578,881 B2 85 86 -continued -continued N N C s if e S HCO S/(N-lN-N N 5 ON S/h (N-N ) N-R N

10 C-S. S. s S-O-if C-N- ) Nu C-s/{ s N- if ON S/( re,NJ

US 9,578,881 B2 89 90 -continued -continued N S- ( -) N-R --N O N-N N

N

N 2 \ O 15 <-- (N- F \ubs’ (N-O-H / H 2O

N / \ O & J (j-N-l 25 )-(N-

30 N-4 \ N O K S. N Z \ N O / H S-cry-O- \-5 H

Z O ----, 50 \-0 H In one aspect, a disclosed compound can have the formula (II):

C rury (II) cror--or ei )-(N- 65 rt (x-R6a R6b -C.le US 9,578,881 B2 91 92 Examplary compound within Formula (II) include, but are -continued not limited to:

5 -(r^- O 10 -(r^- O - Cry- O N- O 15 Ctrl-O C-S. (r)- O O

-(r^- O 25 N -(x-O 30 - (-O

35

six-O -(-)-F F O C--(-)-O O 40 45

-(-)-OF F

50 -(r^- O C- O -(-)-F F O C-s’Orx-O O 55 60 S. Six- O C- O C- O S. re-C 65 s^{rx-O US 9,578,881 B2 93 94 -continued -continued

() - N

/ O CZ -- O C-ty-/ -k-O C crylo 2O h -(r)-O / -(r)- O -(r)-O C-5S/(N- O -O -o-,-O Z-(N--O \ O

C---S/h ( ) -O 50 C-cy--N-N N O F N- ... --Or-O O O ZN H CN -u O C-y-\ N S N-OO O- a 2-(N-O US 9,578,881 B2 95 96 -continued -continued

10

15

N Oreo 25 30 -(r)-O

35 - Cry-O Ciro 40 ) O

45 h CR to 50 -O 55 CS/ \ x -O Certo 60 Or 65 CS/ \ N-O US 9,578,881 B2 97 98 -continued -continued C--- -O -(r^- O

Z N F 15 S. y--O \ N S O F H --Or-O

C-S. (r)--O O 30 C.S. (r^- O O H No H NS O / N N ZN C- s if C-\ N j-NullS O -O 40 S. \N-N Null N -O N-N N Ns Y 45 C-y-7- ) O -O C rulo 50 (r^- O -(N-O C--(r^- S O O 55 Or H --Or-O -(r^- O US 9,578,881 B2 99 -continued CCF -(r^- O 10

O \1 15 Y ai \ N- -O

O aiuC 25 Y (r^- O 30

-iV O Y (r^-O 35 o1 H o1 Z N O C -- -O 45

ZN -O O

C - O 55

60

S. {r^-O 65

US 9,578,881 B2

15 N-O

X-O

35 O

40

55

H-O 65 NO US 9,578,881 B2 105 106 -continued -continued Cl-N-M N O F-N/ C -(r)-O {r^- O C-Ox-C O 19 N/C-cy--O- K N 15 H NNé R N S O F-N/ N O -O-- X-O S. - CC - O O F-/-- try--O-s if C-C, syri S’(x,-O \{x-F H O

( 35 N N S HNN-4 N O S. \ r\-ON

O N 55 N-N H

O H 7 N\ N S O O N N S. / \ O sk (r^- O - C (r-ONH US 9,578,881 B2 107 108 -continued -continued Fy O-N/ C N --,O -O 5 C-cy--ZN O Or - (r-O- - )

ZN l F Cir-O-,y -N -s if -) O l -' (-N- s/{ N- O-4

CS. --O-4- A - 2'-O-H Cur-S. (r-O- CS/ \ rx- N O -' (-O-,-, - re-O- /s ) O C- S O We' so ZN N S. N-O- N- S. \ \ N re-O-S O C -u Ae" / ) O

Cr-C, 60 C-cal CS. \ f -uS Aek 4- )N S O N-N -C C-(-)-O- US 9,578,881 B2 109 110 -continued -continued

15

N

25 O- 30

35

45

N

55

N

O- 65 US 9,578,881 B2 111 112 -continued -continued

C. S O NO N-N N O S. (r^-O- - Cr-Oy CursS- Orn- -O Y- O - (-N- N

ON S- {r^-O 1. 25 Y N O

S. or NJa 30 H 7

S. \ SN-l O s/ N-N N O 40 - ) , 1. Y N yS {r^-O

C- N-NSN-l N -R Nu-o7 \ N S O 50 S. N-O

S. or--O 55 Z (N/ S. 60 stro-,-C). CU-cy--O