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(12) United States Patent (10) Patent No.: US 8,734,869 B2 Enan (45) Date of Patent: May 27, 2014

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(12) United States Patent (10) Patent No.: US 8,734,869 B2 Enan (45) Date of Patent: May 27, 2014 USOO8734869B2 (12) United States Patent (10) Patent No.: US 8,734,869 B2 Enan (45) Date of Patent: May 27, 2014 (54) SYNERGISTIC PEST CONTROL 5,942,214 A 8, 1999 Lucas et al. COMPOSITIONS 6,177.465 B1* 1/2001 Tanaka .......................... 514,535 6,238,682 B1 5, 2001 Klofta et al. 6,849,614 B1* 2/2005 Bessette et al. ................. 514/72 (75) Inventor: Essam Enan, Davis, CA (US) 7,541,155 B2 6/2009 Enan 7,622,269 B2 11/2009 Enan (73) Assignee: TyraTech, Inc., Melbourne, FL (US) 2003/0194454 A1* 10, 2003 Bessette et al. ............... 424,745 2006, 0083763 A1 4/2006 Neale et al. (*) Notice: Subject to any disclaimer, the term of this 2006/0263.403 A1 11/2006 Enan patent is extended or adjusted under 35 2008.0075796 A1 3/2008 Enan U.S.C. 154(b) by 445 days. OTHER PUBLICATIONS (21) Appl. No.: 12/532,604 Colby, S.R., "Calculating Synergistic and Antagonistic Responses of Herbicide Combinations.” Weeds 15(1), p. 20-22, 1967. (22) PCT Filed: Mar. 21, 2008 James, et al. “Field-testing of methyl salicylate for recruitment and retention of beneficial insects in grapes and hops.” Journal of Chemi (86). PCT No.: PCT/US2008/003722 cal Ecology 30(8), p. 1613-1628, 2004. Lynn, D.E., “Development and characterization of insect cell lines.” S371 (c)(1), Cytotechnology 20, p. 3-11, 1996. (2), (4) Date: May 3, 2010 Lynn, D.E., "Methods for maintaining insect cell cultures,” J. Insect Science 2.9, p. 1-6, 2002. (87) PCT Pub. No.: WO2009/038599 Shulaev, et al., “Airborne signalling by methyl salicylate in plant PCT Pub. Date: Mar. 26, 2009 pathogen resistance.” Nature 385, p. 718-721, 1997. U.S. Appl. No. 60/718,570, "Compositions Having Insect Control (65) Prior Publication Data Activity and Methods for Use Thereof.” Sep. 19, 2005. U.S. Appl. No. 60/747,592, “Water-Based Formulation of Insect US 2010/O3O3940 A1 Dec. 2, 2010 Control Compositions and Methods for Production and Use Thereof.” May 18, 2006. Related U.S. Application Data U.S. Appl. No. 60/799.434, “Formulation of Insect-Control Compo (60) Provisional application No. 60/896,427, filed on Mar. sitions Having Residual Activity and Methods for Production and Use Thereof.” May 10, 2006. 22, 2007, provisional application No. 60/896,436, U.S. Appl. No. 60/805.963, “Compositions for Treating Parasitic filed on Mar. 22, 2007, provisional application No. Injections and Methods of Screening for Same.” Jun. 27, 2006. 60/896,430, filed on Mar. 22, 2007, provisional U.S. Appl. No. 60/807,600, "Compositions and Methods for Con application No. 60/987,013, filed on Nov. 9, 2007, trolling Insects.” Jul. 17, 2006. provisional application No. 60/990,912, filed on Nov. PCT International Search Report in corresponding International 28, 2007, provisional application No. 61/023,425, application No. PCT/US08/03722, mailed Sep. 19, 2008. filed on Jan. 24, 2008. * cited by examiner (51) Int. Cl. AOIN 65/00 (2009.01) Primary Examiner — Chris R Tate A6 IK36/00 (2006.01) Assistant Examiner — Russell Fiebig AOIN 43/30 (2006.01) (74) Attorney, Agent, or Firm — Davis Wright Tremaine A6 IK3I/36 (2006.01) LLP (52) U.S. Cl. USPC ............................ 424/778; 424/725; 514/464 (57) ABSTRACT (58) Field of Classification Search Pest control compositions, blends, and formulations are dis None closed. The blends contain, in a synergistic combinations, at See application file for complete search history. least two ingredients such as Lilac Flower Oil, D-Limonene, Thyme Oil, Lime Oil, Black Seed Oil, Wintergreen Oil, Lina (56) References Cited lool, Tetrahydrolinalool, Vanillin, Isopropyl myristate, Piper onal (aldehyde), Geraniol, Geraniol 60, Triethyl Citrate, and U.S. PATENT DOCUMENTS Methyl Salicylate. 4.278,658 A 7/1981 Hooper et al. 4,551480 A 11, 1985 Stiefel et al. 9 Claims, 29 Drawing Sheets U.S. Patent May 27, 2014 Sheet 1 of 29 US 8,734,869 B2 Otal RNA Well anembranes expressing receptor are isolated and used sei in competitive binding assa5. Wwhicle cell used to assay changes in signaling down-stream to the receptor. FIG. 1 U.S. Patent May 27, 2014 Sheet 2 of 29 US 8,734,869 B2 Biogenic amine receptors coupled to intracellular cAMP signaling pathways Once ligand is tightly bound to its receptor, the receptors confirmation will change. This structural change is then transferred ormericooterp -, Physiological responses FIG. 2 U.S. Patent May 27, 2014 Sheet 3 of 29 US 8,734,869 B2 Biogenic amine receptors coupled to intracellular IP3/DAG signaling pathway (C FIG. 3 U.S. Patent May 27, 2014 Sheet 4 of 29 US 8,734,869 B2 Time(secs) FIG. 4 U.S. Patent May 27, 2014 Sheet 5 Of 29 US 8,734,869 B2 ---isd. propyl myris was A ra ex-M s 28 4.333 Tits FIG. 5 U.S. Patent May 27, 2014 Sheet 6 of 29 US 8,734,869 B2 Trre(secs) W. O. As Af a G3 o H6 Mix-Ah -87 85.82 88: 498 FIG. 6 U.S. Patent May 27, 2014 Sheet 7 Of 29 US 8,734,869 B2 em. ... ww a r - - - - - , . A-Auror--- - - - - - - - - - --- " - . " spaw: s chemical Awar. re-or-AAAAs.--arreshi were re. ------. .--na-------------- Y --ee-ear R -- irkasswissive resse--wassax-exseaws.st-stative - we womwv", www.a...seesare rearranme A : 3. sixx-xx-xx-x-xxx8.*.*x war--ories s' woxxyyxx-xxxs ::x:"wir "... - ...vxww.vx Dasas r/at ... - 4 x-axw-rarewwaxes it-ang--Ty Wax Rits-9 W A4 A5 a B3 o E3 E. 5 Wax R Second -774 -1182 44575 2.583 53.8 -15 R2 O080 O.192 01.06 0.01 0.211 O25 FIG. 7 U.S. Patent May 27, 2014 Sheet 8 of 29 US 8,734,869 B2 Natural synergy in endo-parasites, Hymenolepis nana animal study 12O 1OO 80 60 40 U.S. Patent May 27, 2014 Sheet 9 Of 29 US 8,734,869 B2 % Repellency by Formulation 0 1 2 4 6 Hrs. Post-treatment 5-62105 #7-6205 8-62.105 #6-621.05 23 DEE FIG. 9 U.S. Patent May 27, 2014 Sheet 10 of 29 US 8,734,869 B2 6 O 4. O 2 O Treatment with 30% XL 101 Water formulation FIG 10 U.S. Patent May 27, 2014 Sheet 11 of 29 US 8,734,869 B2 y i i ba, B-028t. DM- a Mraw- a B5028 f li- --- re servae t i E essertex-secretacarazresy & - rostrica.k.a. sassist raisi. xxxii. xxxots s --- -- &aw-staxww. W. s r--- - - - - - - - - is a -----DM-------- ifies Y S A * * 4 x-w:x WS 2 E3 a BS X-M 15.1 -158 343 U.S. Patent May 27, 2014 Sheet 12 of 29 US 8,734,869 B2 Time (secs) W. C. F. F3 is Mix- 896 57. 83SOO FIG. 12 U.S. Patent May 27, 2014 Sheet 13 Of 29 US 8,734,869 B2 - axxx w8www.'www.texas:-fixx wroxxsw-ox's Kx - www.xxxar r s way...cxis 3-goa ". Blend 33 i 3. g-, ------------ - - ------------f. : s : k 8 : : FIG. 13 U.S. Patent May 27, 2014 Sheet 14 of 29 US 8,734,869 B2 o l 8 : , ; ----------. ----- -- --- a- - - - - resecs) WaxRs.9 W o E G) G1 WaxStord 88 142S 57 R 2 O3 5 FIG. 14 U.S. Patent May 27, 2014 Sheet 15 Of 29 US 8,734,869 B2 All Treatments, Blend 33 (HL1) and Clothianidin (CL): % Knock Down at 30 seconds Post Application of 1 ml of Test Material: Aedes aegypti. 120 100 OO Treatment FIG. 15 U.S. Patent May 27, 2014 Sheet 16 of 29 US 8,734,869 B2 Middle Dose of Blend 33 (HL1) with Clothianidin (CL): % Knock Down at 30 seconds Post Application of 1 mt of Test Material: Aedes aegypti 120 100 60 HL1 2.5% CL:50 ppm CL:H-1 50 HL1 2.5% CL. 67 CL:HL1 167 H1 2.5% CL 250 CLH1 250 ppm2.5% ppin ppm2.5% ppm ppm2.5% 1:500 ratio 1:150 ratio 1:100 ratio FIG. 16 U.S. Patent May 27, 2014 Sheet 17 Of 29 US 8,734,869 B2 Synergy American Cockroach Clothianidin/Blend 33 (HL1) Synergy Test for American Cockroach 1:100, 1:150, 1:500 Clothianidinf.5% HL-1 FIG. 17 U.S. Patent May 27, 2014 Sheet 18 Of 29 US 8,734,869 B2 4-imid. aclopridi. |Blend-33: p. -------- & -23 g : 1:100: - : ; m (s.d.-- -- risit as will sassi. - sw' -- - - - - - - cre almidacloprid. g m R - - --- !---|-- v. - as 2.x -x a------------ . .swww- w --- - ric- - * www.k - w . i 4. X ""r'ssist Big s Y waxx-r is: T ift-ri- ur-. -Blend 33.-- --- treet is - : X : } jour-for--r i... ----------------e. w.e. scis-rearwalaw assacaw - - was wers his werSet 2S2 133 122 R2 55 08 O8. FIG. 18 U.S. Patent May 27, 2014 Sheet 19 Of 29 US 8,734,869 B2 %. Knock Down at 30 seconds Post Application of 1 ml of Test Material: Aedes aegypti. (Blend 33 = HL1) HL1 2.5% (50 ppm HL150 H1 2.5% - 250 ppm IHL 1 250 HL1 5% - 500 ppm (HL1500 ppm:2.5% ppm:2.5% ppm.5% 15OO ratic 1:1 OO ratio 1:50 ratio FIG. 19 U.S. Patent May 27, 2014 Sheet 20 of 29 US 8,734,869 B2 %. Knock Down at 30 seconds Post Application of 1 ml of Test Material: Drosophila sp.
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