WO2017114052A1 - 高抗逆性的植物生长调节剂及其制法和用途 - Google Patents

高抗逆性的植物生长调节剂及其制法和用途 Download PDF

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WO2017114052A1
WO2017114052A1 PCT/CN2016/107516 CN2016107516W WO2017114052A1 WO 2017114052 A1 WO2017114052 A1 WO 2017114052A1 CN 2016107516 W CN2016107516 W CN 2016107516W WO 2017114052 A1 WO2017114052 A1 WO 2017114052A1
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compound
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朱健康
张玉路
曹民杰
刘雪
王秋华
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Shanghai Institutes for Biological Sciences SIBS of CAS
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Shanghai Institutes for Biological Sciences SIBS of CAS
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Priority to BR112018013314-0A priority patent/BR112018013314B1/pt
Priority to CA3010014A priority patent/CA3010014C/en
Priority to AU2016380736A priority patent/AU2016380736B2/en
Priority to EP16880842.6A priority patent/EP3398940B1/en
Priority to ES16880842T priority patent/ES2923784T3/es
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D265/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom and one oxygen atom as the only ring hetero atoms
    • C07D265/04—1,3-Oxazines; Hydrogenated 1,3-oxazines
    • C07D265/12—1,3-Oxazines; Hydrogenated 1,3-oxazines condensed with carbocyclic rings or ring systems
    • C07D265/14—1,3-Oxazines; Hydrogenated 1,3-oxazines condensed with carbocyclic rings or ring systems condensed with one six-membered ring
    • C07D265/18—1,3-Oxazines; Hydrogenated 1,3-oxazines condensed with carbocyclic rings or ring systems condensed with one six-membered ring with hetero atoms directly attached in position 2
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
    • A01N43/40—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom six-membered rings
    • A01N43/42—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom six-membered rings condensed with carbocyclic rings
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/48—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with two nitrogen atoms as the only ring hetero atoms
    • A01N43/54—1,3-Diazines; Hydrogenated 1,3-diazines
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/72—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms
    • A01N43/86—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms six-membered rings with one nitrogen atom and either one oxygen atom or one sulfur atom in positions 1,3
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D215/20—Oxygen atoms
    • C07D215/22—Oxygen atoms attached in position 2 or 4
    • C07D215/227—Oxygen atoms attached in position 2 or 4 only one oxygen atom which is attached in position 2
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
    • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D215/38—Nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/70—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings condensed with carbocyclic rings or ring systems
    • C07D239/72—Quinazolines; Hydrogenated quinazolines
    • C07D239/78—Quinazolines; Hydrogenated quinazolines with hetero atoms directly attached in position 2
    • C07D239/80—Oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D263/00—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings
    • C07D263/52—Heterocyclic compounds containing 1,3-oxazole or hydrogenated 1,3-oxazole rings condensed with carbocyclic rings or ring systems
    • C07D263/54—Benzoxazoles; Hydrogenated benzoxazoles
    • C07D263/58—Benzoxazoles; Hydrogenated benzoxazoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached in position 2
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/07—Optical isomers

Definitions

  • the present invention relates to the field of botany, and in particular to a plant growth regulator having high stress resistance, a process for its preparation and use.
  • Abscisic Acid is a key factor in balancing the metabolism of endogenous hormones and related active substances in plants. It has the ability to promote the balanced absorption of water, fertilizer and coordination of metabolism in plants, and can effectively regulate root/crown and vegetative growth of plants. And reproductive growth plays an important role in improving the quality and yield of crops.
  • ABA Abscisic Acid
  • exogenous ABA can cause rapid closure of leaf stomata, inhibit transpiration, and can be used for flower preservation, or to prevent wilting during transportation of crop seedlings.
  • ABA can also control flower bud differentiation, regulate flowering, and has great application value in flower gardening.
  • ABA can improve the growth of crops in low temperature, drought, spring cold, salt and other adverse growth environments. Therefore, ABA has a wide range of applications and can be used in lawns, farmland, gardens, and especially in water-deficient areas such as the western region, which is of great significance for the development of China's agricultural industry.
  • a first aspect of the invention provides a compound of formula I, or a salt thereof, or an optical isomer thereof, or a racemate thereof, or a solvate thereof, or a precursor thereof,
  • R 1 is H, halogen, C 1 -C 3 alkyl, or C 1 -C 3 haloalkyl
  • R 2 is H, halogen, C 1 -C 3 alkyl, or C 1 -C 3 haloalkyl
  • R 3 is H, halogen, C 1 -C 3 alkyl, or C 1 -C 3 haloalkyl
  • R 4 is H, halogen, C 1 -C 3 alkyl, or C 1 -C 3 haloalkyl
  • R 5 is halogen, C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, SF 5 or C 3 -C 8 cycloalkyl;
  • R 6 is a substituted or unsubstituted C 1 -C 7 alkyl group, a substituted or unsubstituted C 2 -C 7 alkenyl group, a substituted or unsubstituted C 2 -C 7 alkynyl group, a substituted or unsubstituted C a 3- C 7 cycloalkyl group, or a substituted or unsubstituted -R a -OR b , wherein R a is a C 1 -C 2 alkylene group and R b is a H, C 1 -C 3 alkyl group; Substituted as having one or more substituents selected from the group consisting of halogen, -OR b , -CN, -N(R b ) 2 , and nitro;
  • R 7 is selected from the group consisting of H, substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 2 -C 6 alkenyl, substituted or unsubstituted C 2 -C 6 alkynyl, Substituted or unsubstituted C 3 -C 8 cycloalkyl, substituted or unsubstituted C 5 -C 10 heterocyclic group, R c -C(O)-, -OR b , -CN, and -N(R b 2 ; R c is selected from the group consisting of a hydroxyl group, a mercapto group, a substituted or unsubstituted C 1 -C 6 alkyl group, a substituted or unsubstituted C 1 -C 6 alkoxy group; wherein the heterocyclic group contains 1 To 2 heteroatoms selected from N, O, S, and the substitutions are one or more substituents selected
  • R 8 , R 9 and R 10 are each independently selected from the group consisting of:
  • R 11 is H, C 1 -C 3 alkyl, or C 1 -C 3 haloalkyl
  • X is CR 12 , NR 13 , O, or S, wherein R 12 is selected from the group consisting of H, halogen, C 1 -C 3 alkyl, C 2 -C 3 alkenyl, C 2 -C 3 alkynyl , C 1 -C 3 haloalkyl, or a combination thereof; R 13 is absent or selected from the group consisting of H, halogen, C 1 -C 3 alkyl, C 2 -C 3 alkenyl, C 2 -C 3 An alkynyl group, a C 1 -C 3 haloalkyl group, or a combination thereof;
  • R 6 is C 1 -C 7 alkyl, C 2 -C 7 alkenyl, C 2 -C 7 alkynyl, C 3 -C 7 cycloalkyl, or -R a - OR b , wherein R a is a C 1 -C 2 alkylene group and R b is H, C 1 -C 3 alkyl.
  • R 7 is selected from the group consisting of substituted or unsubstituted C 1 -C 6 alkyl, substituted or unsubstituted C 2 -C 6 alkenyl, substituted or unsubstituted C 2 -C 6 alkynyl, substituted or unsubstituted C 3 -C 8 cycloalkyl, substituted or unsubstituted C 5 -C 10 heterocyclyl, R c -C(O)-, -OR b , -CN, and -N(R b ) 2 ;
  • R c is selected from the group consisting of a hydroxyl group, a mercapto group, a substituted or unsubstituted C 1 -C 6 alkyl group, a substituted or unsubstituted C 1 -C 6 alkoxy group;
  • the heterocyclic group contains 1 to 2 hetero atoms selected from N, O, S, and the substitution is one or more substituents selected from
  • R 7 is H.
  • R 8 , R 9 , and R 10 are simultaneously H.
  • R 13 is selected from the group consisting of H, halogen, C 1 -C 3 alkyl, C 2 -C 3 alkenyl, C 2 -C 3 alkynyl, C 1 -C 3
  • the halogen includes F, Cl, Br or I.
  • the halogen is F.
  • the compound has the structure of Formula Ia:
  • R 1 - R 10 and m are as defined above.
  • the compound has the structure of formula Ib:
  • the compound has the structure of formula Ic:
  • R 1 - R 10 and m are as defined above.
  • the compound has the structure of formula Id:
  • R 1 - R 10 , R 12 , m are as defined above.
  • R 1 , R 2 , R 3 and R 4 are all H.
  • 1 , 2 , 3 or 4 of R 1 , R 2 , R 3 and R 4 are halogen.
  • the halogen includes F, Cl, Br or I.
  • the halogen is F.
  • R 1 , R 2 , R 3 and R 4 are F.
  • R 5 is C 1 -C 3 alkyl, C 1 -C 3 haloalkyl, SF 5 or C 3 -C 6 cycloalkyl.
  • R 5 is methyl or cyclopropyl.
  • R 6 is a substituted or unsubstituted C 1 -C 5 alkyl group, a substituted or unsubstituted C 2 -C 5 alkenyl group, a substituted or unsubstituted C 2 -C 5 alkynyl group.
  • R a is a C 1 -C 2 alkylene group and R b is H, C 1 - C 3 alkyl; and the substitution is one or more substituents selected from the group consisting of halogen, -OR b , -CN, -N(R b ) 2 , or a nitro group.
  • R 6 is n-propyl, ethyl, isopropyl, isobutyl, or fluoro-n-propyl.
  • R 6 is a C 3 alkyl group, a C 3 alkenyl group, or a C 3 alkynyl group.
  • R 6 is n-propyl
  • R 7 is selected from the group consisting of H, substituted or unsubstituted C 1 -C 4 alkyl, substituted or unsubstituted C 2 -C 4 alkenyl, substituted or unsubstituted C 2 -C 4 alkynyl, substituted or unsubstituted C 3 -C 6 cycloalkyl, substituted or unsubstituted C 5 -C 8 heterocyclic group, R c -C(O)-, -OH, -CN, and -NH 2 ;
  • R c is selected from the group consisting of a hydroxyl group, a mercapto group, a substituted or unsubstituted C 1 -C 4 alkyl group, a substituted or unsubstituted C 1 -C 4 alkoxy group; wherein the heterocyclic group contains 1 to 2 hetero atoms selected from N, O, S, and the substitution is one or more substituents selected from the group consisting of
  • R 8 , R 9 and R 10 are each independently selected from the group consisting of:
  • R c is selected from the group consisting of: a hydroxy, decyl, substituted or unsubstituted C 1 -C 4 alkyl, substituted or unsubstituted C 1 -C 4 alkoxy; wherein the substituent is one or more substituents selected from the group consisting of: Halogen, -OH, -CN, -NH 2 , and nitro.
  • R 7 is selected from the group consisting of H, substituted or unsubstituted C 1 -C 4 alkyl, substituted or unsubstituted C 2 -C 4 alkenyl, substituted or unsubstituted C 2 -C 4 alkynyl, substituted or unsubstituted C 3 -C 6 cycloalkyl, substituted or unsubstituted C 5 -C 8 heterocyclic group, R c -C(O)-, -OH, -CN, and -NH 2 ;
  • R c is selected from the group consisting of a hydroxyl group, a mercapto group, a substituted or unsubstituted C 1 -C 4 alkyl group, a substituted or unsubstituted C 1 -C 4 alkoxy group; wherein the heterocyclic group contains a hetero atom selected from N, O, S, and the substitution is one or more substituents selected from the group consisting of hal
  • R 8 , R 9 and R 10 are each independently selected from the group consisting of:
  • R c is selected from the group consisting of: a hydroxy, decyl, substituted or unsubstituted C 1 -C 4 alkyl, substituted or unsubstituted C 1 -C 4 alkoxy; wherein the substituent is one or more substituents selected from the group consisting of: Halogen, -OH, -CN, -NH 2 , and nitro.
  • R 7 is selected from the group consisting of substituted or unsubstituted C 1 -C 4 alkyl, substituted or unsubstituted C 2 -C 4 alkenyl, substituted or unsubstituted C 2 -C 4 alkynyl, substituted or unsubstituted C 3 -C 6 cycloalkyl, substituted or unsubstituted C 5 -C 8 heterocyclic group, R c -C(O)-, -OH, -CN, and -NH 2 ; R c is selected from the group consisting of a hydroxyl group, a mercapto group, a substituted or unsubstituted C 1 -C 4 alkyl group, a substituted or unsubstituted C 1 -C 4 alkoxy group; wherein the heterocyclic group contains 1 to 2 heteroatoms selected from N, O, S, and said substituents having one or more substituents selected from the group consisting of
  • R 8 , R 9 and R 10 are each independently selected from the group consisting of:
  • R c is selected from the group consisting of: a hydroxy, decyl, substituted or unsubstituted C 1 -C 4 alkyl, substituted or unsubstituted C 1 -C 4 alkoxy; wherein the substituent is one or more substituents selected from the group consisting of: Halogen, -OH, -CN, -NH 2 , and nitro.
  • R 7 is selected from the group consisting of substituted or unsubstituted C 1 -C 4 alkyl, substituted or unsubstituted C 2 -C 4 alkenyl, substituted or unsubstituted C 2 -C 4 alkynyl, substituted or unsubstituted C 3 -C 6 cycloalkyl, substituted or unsubstituted C 5 -C 8 heterocyclic group, R c -C(O)-, -OH, -CN, and -NH 2 ; R c is selected from the group consisting of a hydroxyl group, a mercapto group, a substituted or unsubstituted C 1 -C 4 alkyl group, a substituted or unsubstituted C 1 -C 4 alkoxy group; wherein the heterocyclic group contains 1 a hetero atom selected from N, O, S, and the substitution is one or more substituents selected from the group consisting of halogen
  • R 8 , R 9 and R 10 are each independently selected from the group consisting of:
  • R c is selected from the group consisting of: a hydroxy, decyl, substituted or unsubstituted C 1 -C 4 alkyl, substituted or unsubstituted C 1 -C 4 alkoxy; wherein the substituent is one or more substituents selected from the group consisting of: Halogen, -OH, -CN, -NH 2 , and nitro.
  • R 7 , R 8 , R 9 and R 10 are the specific groups corresponding to the specific compounds in the examples of the present application.
  • the compound is selected from the group consisting of:
  • the compound is selected from the group consisting of:
  • a second aspect of the invention provides a use of a compound of formula I according to the first aspect of the invention, or a salt thereof, or an optical isomer thereof, or a racemate thereof, or a solvate thereof, or a precursor thereof,
  • a compound of formula I for the preparation of an agricultural preparation or composition for (i) enhancing plant stress resistance; (ii) preparing an agonist of an ABA receptor; and/or (iii) preparing a seed germination inhibitor.
  • the agonist promotes the interaction of the ABA receptor PYL protein with the PP2C protein phosphatase.
  • the agricultural formulation or composition is for one or more of the following uses:
  • the stress resistance is ABA-related abiotic stress resistance.
  • the stress resistance is selected from the group consisting of drought resistance, cold tolerance, salt and alkali resistance, osmotic pressure resistance, heat resistance, or a combination thereof.
  • the plant is a plant comprising a PYR/PYL family ABA receptor.
  • the plant comprises moss, fern, gymnosperm, monocot, and dicot.
  • the plant comprises an agricultural plant, a horticultural plant, a forestry plant.
  • the plant comprises a woody plant, a herbaceous plant.
  • the plant comprises intact plants, organs (such as roots, stems, leaves, branches, flowers, fruits, seeds), tissues (such as callus), or cells.
  • organs such as roots, stems, leaves, branches, flowers, fruits, seeds
  • tissues such as callus
  • the plant is selected from the group consisting of Gramineae, Compositae, Liliaceae, Cruciferae, Rosaceae, Leguminosae, Tea, Sycamore, Pine, Walnut, Piper, Magnoliaceae, Rhododendron, Actinidia, Vitis, Begonia, Bromeliaceae, Ginkgo, Star Anise, Zingiberaceae, Pomegranate, Apocynaceae, Polygonaceae, Rutaceae, Solanaceae, Cypress, Holly, palm plant, or a combination thereof.
  • the plant is selected from the group consisting of Arabidopsis thaliana, tobacco, cotton, lettuce, rice, wheat, corn, peanuts, sorghum, oats, rye, sugar cane, soybeans, potatoes, buckwheat, pepper, Grape, pear, apple, banana, ginseng, tomato, pepper, eggplant, broccoli, Chinese cabbage, rape, cucumber, watermelon, onion, sunflower, lily, rose, chrysanthemum, peony, carnation, eucalyptus, sycamore, pine, or a combination thereof .
  • a third aspect of the invention provides an agricultural preparation comprising:
  • the component (i) is contained in the agricultural preparation in an amount of from 0.1 to 1000 ⁇ M, preferably from 1 to 200 ⁇ M, more preferably from 5 to 100 ⁇ M.
  • the agricultural preparation contains 0.0001 to 99% by weight, preferably 0.1 to 90% by weight, of the component (i), based on the total weight of the agricultural preparation.
  • the agricultural preparation further comprises an additional drought-tolerant (such as a drought-resistant seed coating, a drought-resistant water-retaining agent, or a drought-resistant spray) or other agricultural active ingredient.
  • an additional drought-tolerant such as a drought-resistant seed coating, a drought-resistant water-retaining agent, or a drought-resistant spray
  • the agricultural active ingredient is selected from the group consisting of fungicides, herbicides, insecticides, nematicides, insecticides, plant activators, synergists, plant growth regulators, Acaricide.
  • the agricultural formulation further comprises a surfactant (such as a cationic, anionic, amphoteric, or nonionic surfactant).
  • a surfactant such as a cationic, anionic, amphoteric, or nonionic surfactant.
  • the dosage form of the agricultural preparation is selected from the group consisting of a solution, an emulsion, a suspension, a powder, a foaming agent, a paste, a granule, an aerosol, or a combination thereof.
  • a fourth aspect of the invention provides a method of enhancing plant stress resistance, which comprises administering to a plant a compound of formula I according to the first aspect of the invention, or a salt thereof, or an optical isomer thereof, or a racemic thereof The body, or a solvate thereof, or a precursor thereof, or the agricultural preparation of the third aspect of the invention.
  • the administration is selected from the group consisting of spraying or irrigation.
  • the dosage is from 2 to 100 g/ha, preferably from 4 to 80 g/ha, more preferably Ground, 6-60g / ha.
  • the dose to be administered is 1-5000 ⁇ g/plant, preferably 10-2500 ⁇ g/plant, more preferably 20-1000 ⁇ g/plant.
  • a process for the preparation of a compound of the formula I or a salt thereof comprising the steps of:
  • the inert solvent is selected from the group consisting of N,N-dimethylformamide (DMF), dichloromethane (DCM), acetonitrile (ACN), or a combination thereof.
  • DMF N,N-dimethylformamide
  • DCM dichloromethane
  • ACN acetonitrile
  • reaction is carried out in the presence of an acid binding agent.
  • the acid binding agent is selected from the group consisting of potassium carbonate (K 2 CO 3 ), triethylamine (Et 3 N), pyridine (Py), or a combination thereof.
  • the reaction temperature is 0 to 150 ° C (or reflux temperature), preferably 10 to 60 ° C, more preferably 20 to 40 ° C.
  • the reaction time is from 0.1 to 72 hours, more preferably from 1 to 24 hours, more preferably from 8 to 20 hours, more preferably from 4 to 12 hours.
  • X is O, and Is a single button.
  • the compound I-S2 is prepared by the following method:
  • R 1 , R 2 , R 3 , R 4 and R 5 are as defined in the first aspect of the invention, and X 2 is a leaving group such as Cl, Br or I.
  • the inert solvent is selected from the group consisting of ethanol, acetonitrile, tetrahydrofuran, or a combination thereof.
  • reaction is carried out under acidic conditions.
  • the acid is selected from the group consisting of hydrochloric acid, hydrobromic acid, or a combination thereof.
  • the reaction temperature is 0 to 150 ° C (or reflux temperature), preferably 10 to 50 ° C, more preferably 15 to 25 ° C.
  • the reaction time is from 0.1 to 72 hours, more preferably from 1 to 24 hours, more preferably from 2 to 12 hours.
  • Figure 1 shows a plurality of compounds of the invention, including 0224 (Figure 1a), 0304 ( Figure 1b), 0706, 0708, 0713, 0715 (Figure 1c), 1028c ( Figure 1d), 0428 (1e), 1022B (1f) and NC0F4 (1g) binds to the Arabidopsis PYL2 receptor-HAB1 complex, thereby inhibiting the activity of the protein phosphatase HAB1. And at lower concentrations, the above compounds have inhibitory effects, and most compounds have better or significantly better inhibitory effects than ABA.
  • Figure 2 shows the presence of a number of compounds of the invention as a PYL receptor agonist with a dose dependent effect.
  • Figure 3 shows the effect of 0224 compound and ABA on seed germination of Col-0 and pyr1; pyl1; pyl4 triple mutant at a concentration of 1 ⁇ M.
  • Col-0 was sown in the left half of each dish, and pyr1; pyl1; pyl4 triple mutant was seeded in the right half.
  • the picture is obtained from pyr1; pyl1; pyl4 triple mutant seeds after 4 days of germination (after 6 days of sowing).
  • DMSO treatment was used as a control group.
  • Figure 4 shows that treatment of the compound 0224, 0304, 0706, 0715 or 0428 of the present invention significantly reduced the transpiration rate of the Arabidopsis leaf surface, resulting in an increase in leaf surface temperature.
  • Figure 4a shows that after treatment with 5 ⁇ M ABA or Compound 0224, the leaf surface temperature increased significantly compared with the control (DMSO) treatment, and the compound 0224 lasted longer;
  • Figure 4b showed that the 5 ⁇ M compound 0304 treatment, the leaf surface temperature was compared with the control Group (DMSO) treatment increased significantly, while the compound 0224 concentration decreased to 2 ⁇ M or 1 ⁇ M, the leaf surface temperature was significantly increased compared with the control (DMSO) treatment, and the effect decreased stepwise, indicating the inhibition of compound 0224 on foliar transpiration There was a concentration-dependent effect in the effect;
  • Figure 4c shows that after treatment with 5 ⁇ M of compound 0706 or 0715, the leaf surface temperature was significantly increased compared to the control (DMSO) treatment, with a duration comparable to 0224;
  • Figure 5 shows that the treatment of compounds 0224 and 0304 of the present invention significantly reduced the transpiration rate of soybean leaf surface, resulting in an increase in leaf surface temperature. Soybean plants after 14 days of sowing were stopped from watering while spraying the compound 0224, 0304 or ABA of the present invention, and 20 ⁇ M of the compound compared with the control group (DMSO). 0224 or 0304 can significantly reduce the transpiration rate of soybean leaf surface, and the inhibition effect is better than the same concentration of ABA.
  • Figure 6 shows that treatment of compound 0224 of the present invention significantly reduced the transpiration rate of cotton leaf surface, resulting in elevated leaf surface temperatures.
  • 20 ⁇ M of compound 0224 can significantly reduce the transpiration rate of cotton leaf surface, and the inhibition effect is better.
  • the same concentration of ABA is better.
  • Figure 7 shows the results of soil drought experiments in Arabidopsis.
  • Arabidopsis thaliana was photographed before drought and four weeks after drought.
  • Arabidopsis thaliana in the control group (DMSO treatment) had dried up after four weeks of drought, and Arabidopsis thaliana treated with compound 0224, 0706 or 0715 still grew normally.
  • FIG 8 shows the results of soil drought experiments in soybeans.
  • the soybean in Figure 5 was rehydrated after 6 days of drought, and the photograph shows the growth of soybeans after one day of rehydration.
  • the growth of soybeans treated with compound 0224 or 0304 was significantly better than that of the control (DMSO) or the same concentration of ABA treated soybeans.
  • Figure 9 shows the results of soil drought experiments in cotton.
  • the cotton in Fig. 6 was rehydrated after 6 days of drought, during which the compound 0224 or ABA of the present invention was sprayed every 3 days, and the photograph showed the growth of cotton before rehydration and after one day of rehydration.
  • the cotton treated with compound 0224 was significantly better than the control (DMSO) or the same concentration of ABA treated cotton.
  • Figure 10 shows the transcriptional level changes of ABA-induced stress-related genes in wild-type Arabidopsis thaliana after treatment with 10 ⁇ M of the present compound 1022B for 6 hours.
  • DMSO and the same concentration of ABA were negative and positive control, respectively.
  • the results showed that the transcription levels of the four stress-related genes induced by the compound 1022B of the present invention were higher than those of the ABA.
  • Figures 11a and 11b show the two-dimensional structure of ABA(a) or a compound of the invention (0428) (b) interacting with multiple amino acid residues within the PYL2-HAB1 complex pocket structure, respectively.
  • the figure shows the water molecule, the nitrogen atom, the oxygen atom and the halogen atom, the dashed line represents the hydrogen bond, and the number on it represents the distance between two atoms/molecule (in angstroms, ).
  • the compound 0428 of the present invention forms a plurality of hydrogen bonds with the amino acid residues in the PYL2 pocket structure, except that the formation of these hydrogen bonds does not require water molecule mediated, which contributes to the compound 0428 and The PYL2-HAB1 complex is more tightly bound.
  • Figure 12 shows the results of soil drought experiments in Arabidopsis. Wild-type Arabidopsis plants (Col-0) grown for 3 weeks in a short-day environment were stopped from watering and sprayed with 5 ⁇ M ABA or the compound 0428 of the present invention. The pictures show the growth of the plants on the day of the first spraying of the compound and after 14 days, and the plants sprayed with the DMSO solution served as a negative control. The results showed that the plant growth of the compound 4428 was superior to that of the control group and the ABA-sprayed plants.
  • Figure 13 shows dose response curves of compounds of the invention (e.g., compound 0428) and ABA of soybean GmPYL6 and rice OsPYL2 receptor agonists.
  • Compound 0428 can promote the interaction of Arabidopsis protein phosphatase HAB1 with soybean GmPYL6 or rice OsPYL2, and this interaction has a dose-dependent effect.
  • Figures 14a and 14b show the results of soil drought experiments for soybean and corn, respectively.
  • the corn of the small bell stage and the soybean plants of the three groups of three-leaf stage were selected, and the compound of the present invention (such as compound 0428) was sprayed on the first day and the second day after the start of the drought, and the photograph showed that the corn was treated for four days in drought.
  • the concentration of compound 0428 in the experiment was 50 ⁇ M.
  • the growth of corn and soybean treated with compound 0428 was significantly better than that of the control group.
  • Figure 15 shows the effect of compound 0428 and ABA on seed germination of Col-0 and pyr1; pyl1; pyl4 triple mutants at a concentration of 2 ⁇ M. Sowing the left half of each dish with Col-0, sowing the right half Pyr1; pyl1; pyl4 triple mutant. The photographs were taken for pyr1; pyl1; pyl4 triple mutant seeds after 7 days of germination (after 9 days of sowing). DMSO treatment was used as a control group.
  • Figure 16 shows that treatment of compound 0224 of the present invention significantly reduced the transpiration rate of wheat leaf surface, resulting in an increase in leaf surface temperature.
  • the wheat plants after 18 days of sowing were stopped from watering while spraying the compound 0224 of the present invention, and 100 ⁇ M of the compound 0224 significantly reduced the transpiration rate of the wheat leaf surface as compared with the control group (DMSO).
  • DMSO control group
  • Figure 17 shows the results of soil drought experiments in wheat.
  • the wheat in Fig. 16 was photographed after 6 days of drought, and the photograph showed that the growth of wheat treated with 100 ⁇ M of compound 0224 or ABA was significantly superior to that of the control group (DMSO).
  • the inventors have for the first time developed a class of ABA substitutes (compounds of the present invention) having high abscisic acid (ABA) activity through extensive and intensive research.
  • the compounds of the present invention are capable of significantly enhancing a variety of stress resistance of plants (e.g., drought resistance, cold tolerance, etc.).
  • the compound of the present invention has a simple preparation method, and has the advantages of excellent environmental friendliness and rapid action, and therefore has broad application prospects.
  • the present invention has been completed on this basis.
  • substituted or unsubstituted means that the group may be unsubstituted or that H in the group is one or more (eg 1-10, preferably 1-5) More preferably 1-3, optimally, 1-2) substituents are substituted.
  • substituted or “substituted” means that the group has one or more (preferably 1-6, more preferably 1-3) substituents selected from the group consisting of: Halogen, hydroxy, -NH 2 , nitro, -CN, C 1 -C 4 alkyl, C 1 -C 4 haloalkyl, C 1 -C 4 alkoxy, C 3 -C 6 cycloalkyl, C 1 -C 3 carboxyl group, C 2 -C 4 alkenyl group, C 2 -C 4 alkynyl group.
  • substituents selected from the group consisting of: Halogen, hydroxy, -NH 2 , nitro, -CN, C 1 -C 4 alkyl, C 1 -C 4 haloalkyl, C 1 -C 4 alkoxy, C 3 -C 6 cycloalkyl, C 1 -C 3 carboxyl group, C 2 -C 4 alkenyl group, C 2 -C 4 alkynyl group
  • C 1 -C 7 alkyl refers to a straight or branched alkyl group having from 1 to 7 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, iso Butyl, sec-butyl, tert-butyl, or the like.
  • the term includes substituted or unsubstituted C 1 -C 7 alkyl, unless otherwise specified.
  • C 1 -C 6 alkyl refers to a straight or branched alkyl group having from 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, iso Butyl, sec-butyl, tert-butyl, or the like.
  • the term includes a substituted or unsubstituted C 1 -C 6 alkyl group, unless otherwise specified.
  • C 1 -C 4 alkyl refers to a straight or branched alkyl group having from 1 to 4 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, iso Butyl, sec-butyl, tert-butyl, or the like.
  • the term includes substituted or unsubstituted C 1 -C 4 alkyl, unless otherwise specified.
  • C 1 -C 3 alkyl refers to a straight or branched alkyl group having from 1 to 3 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, or the like. group.
  • the term includes substituted or unsubstituted C 1 -C 3 alkyl, unless otherwise specified.
  • C 1 -C 2 alkylene refers to a divalent hydrocarbon group having 1-2 carbon atoms, such as a methylene group, an ethylene group, or the like.
  • the term includes a substituted or unsubstituted C 1 -C 2 alkylene group, unless otherwise specified.
  • C 2 -C 7 alkenyl refers to a straight or branched alkenyl group having 2 to 7 carbon atoms, such as ethenyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or the like.
  • the term includes a substituted or unsubstituted C 2 -C 7 alkenyl group, unless otherwise specified.
  • C 2 -C 6 alkenyl refers to a straight or branched alkenyl group having 2 to 6 carbon atoms, such as ethenyl, allyl, 1-propenyl, isopropenyl, 1-butenyl, 2-butenyl, or the like.
  • the term includes a substituted or unsubstituted C 2 -C 6 alkenyl group, unless otherwise specified.
  • C 2 -C 3 alkenyl refers to a straight or branched alkenyl group having 2 to 3 carbon atoms, such as vinyl, allyl, 1-propenyl, isopropenyl or Similar group.
  • the term includes a substituted or unsubstituted C 2 -C 3 alkenyl group, unless otherwise specified.
  • C 2 -C 7 alkynyl group means a straight or branched alkynyl chain having 2-7 carbon atoms, e.g. ethynyl, propynyl, or the like.
  • the term includes a substituted or unsubstituted C 2 -C 7 alkynyl group, unless otherwise specified.
  • C 2 -C 6 alkynyl group means a straight or branched alkynyl chain having 2-6 carbon atoms, e.g. ethynyl, propynyl, or the like.
  • the term includes a substituted or unsubstituted C 2 -C 6 alkynyl group, unless otherwise specified.
  • C 2 -C 3 alkynyl group means a straight or branched alkynyl chain having 2-3 carbon atoms, e.g. ethynyl, propynyl, or the like.
  • the term includes a substituted or unsubstituted C 2 -C 3 alkynyl group, unless otherwise specified.
  • C 3 -C 8 cycloalkyl refers to a cyclic alkyl group having from 3 to 8 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or Similar group.
  • the term includes a substituted or unsubstituted C 3 -C 8 cycloalkyl group, unless otherwise specified.
  • C 3 -C 7 cycloalkyl refers to a cyclic alkyl group having 3 to 7 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or Similar group.
  • the term includes a substituted or unsubstituted C 3 -C 7 cycloalkyl group, unless otherwise specified.
  • C 3 -C 6 cycloalkyl refers to a cyclic alkyl group having 3-6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or the like. The term includes a substituted or unsubstituted C 3 -C 6 cycloalkyl group, unless otherwise specified.
  • C 5 -C 10 heterocyclyl refers to a saturated, partially saturated or unsaturated group (but not aromatic) having a single ring or a fused ring (including a bridged ring system and a spiro ring).
  • one or more of the rings may be a cycloalkyl, aryl or heteroaryl group, As long as the junction passes through a non-aromatic ring.
  • the term includes substituted or unsubstituted heterocyclic groups.
  • C 5 -C 8 heterocyclyl refers to a saturated, partially saturated or unsaturated group (but not aromatic) having a single ring or a fused ring (including a bridged ring system and a spiro ring). a system having 5 to 8 carbon atoms and 1 to 2 heteroatoms selected from nitrogen, sulfur or oxygen), wherein in the fused ring system, one or more of the rings may be a cycloalkyl, aryl or heteroaryl group, As long as the junction passes through a non-aromatic ring.
  • the term includes substituted or unsubstituted heterocyclic groups.
  • C 1 -C 3 haloalkyl refers to a straight or branched alkyl group having from 1 to 3 carbon atoms in which hydrogen is substituted by one or more halogens, for example, a halomethyl group. , haloethyl, halopropyl, haloisopropyl, or the like.
  • the term includes a substituted or unsubstituted C 1 -C 3 haloalkyl group, unless otherwise specified.
  • C 1 -C 6 alkoxy refers to a group having the structure "(C 1 -C 6 alkyl)-O-, for example, CH 3 -O-, C 2 H 5 -O -, C 3 H 7 -O-, (CH 3 ) 2 CH-O-, nC 4 H 9 -O-, tC 4 H 9 -O-, or the like. The term is used unless otherwise specified. A substituted or unsubstituted C 1 -C 6 alkoxy group is included.
  • C 1 -C 4 alkoxy refers to a group having the structure "(C 1 -C 4 alkyl)-O-, for example, CH 3 -O-, C 2 H 5 -O -, C 3 H 7 -O-, (CH 3 ) 2 CH-O-, nC 4 H 9 -O-, tC 4 H 9 -O-, or the like. The term is used unless otherwise specified. A substituted or unsubstituted C 1 -C 4 alkoxy group is included.
  • halogen means fluoro, chloro, bromo, or iodo, preferably fluoro and chloro, most preferably fluoro.
  • halo refers to a group substituted by the same or different one or more of the above halogen atoms, and may be partially or fully halogenated, such as trifluoromethyl, pentafluoroethyl, heptafluoro. Isopropyl, or a similar group.
  • the compounds of the invention may contain one or more asymmetric centers and thus occur as racemates, racemic mixtures, single enantiomers, diastereomeric compounds and single diastereomers.
  • the asymmetric center that can exist depends on the nature of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers, and all possible optical isomers and diastereomeric mixtures and pure or partially pure compounds are included within the scope of the invention.
  • the invention includes all isomeric forms of the compounds.
  • the terms “compound of the invention”, “analog of ABA of the invention”, “compound of formula I” are used interchangeably and refer to a compound having the structure of formula I. Furthermore, the term also includes salts, optical isomers, racemates, solvates (eg, hydrates), and/or precursors of the compounds of Formula I,
  • R 1 - R 10 , R 11 , m, X The definition is as described above.
  • the compound has the structure of Formula Ia:
  • R 1 - R 10 and m are as defined above.
  • the compound has the structure of formula Ib:
  • the compound has the structure of formula Ic:
  • R 1 - R 10 and m are as defined above.
  • the compound has the structure of formula Id:
  • R 1 - R 10 , R 12 , m are as defined above.
  • each reaction is mostly carried out in an inert solvent at 0 ° C to 150 ° C (or reflux temperature) (preferably, 10-60 ° C, or 20-40 ° C) for a period of time. (eg 0.1-72 hours, preferably 2-20 hours).
  • room temperature means 4-35 ° C, preferably 20-30 ° C.
  • the compounds of formula I of the present invention can be prepared by the following schemes and exemplary methods described in the Examples and related publications used by those skilled in the art.
  • the methods of preparing the compounds of Formula Ia of the present invention can include, but are not limited to, the following schemes.
  • step 1 in a inert solvent (such as tetrahydrofuran), the compound of formula I-1 is first reacted with N,N'-carbonyldiimidazole (CDI) at a certain temperature (such as 20-40 ° C) for a period of time to form Formula I-2 Compound.
  • a inert solvent such as tetrahydrofuran
  • Step 2 In the presence of a base such as sodium hydride, the compound of formula 1-2 is reacted with iodine propane at a temperature (eg 20-40 ° C) in an inert solvent such as N,N-dimethylformamide. The reaction is continued for a period of time to form a compound of formula 1-3.
  • a base such as sodium hydride
  • Step 3 The compound of formula 1-3 is reacted with potassium nitrate at a temperature (e.g., 0-10 ° C) for a period of time in the presence of an acid such as sulfuric acid to form a compound of formula 1-4.
  • a temperature e.g., 0-10 ° C
  • Step 4 The compound of the formula I-4 is subjected to a reduction reaction in an inert solvent (e.g., methanol) using palladium on carbon as a catalyst at a temperature (e.g., 20 to 40 ° C) to form a compound of the formula I-5.
  • an inert solvent e.g., methanol
  • palladium on carbon e.g., palladium on carbon
  • Step 5 Compounds of formula I-SS1 (eg 2,3,5,6-tetrafluoro-4-methylbenzyl bromide or 2,3,5,6-tetrafluoro-) in an inert solvent such as ethanol or acetonitrile 4-methylbenzyl chloride) is reacted with thiourea to form a reaction product; then, the reaction product and sodium chlorite are at a certain temperature in an inert solvent such as acetonitrile in the presence of an acid such as concentrated hydrochloric acid. The reaction is carried out for a period of time (e.g., 15-25 ° C) to form a compound of formula I-S2.
  • an inert solvent such as ethanol or acetonitrile 4-methylbenzyl chloride
  • Step 6 The compound of the formula I-5 is reacted with the compound of the formula I-S2 at a certain temperature (for example, 20-50 ° C) for a while in an inert solvent such as DMF in the presence of an acid binding agent such as potassium carbonate. Thereby obtaining a compound of the formula Ia.
  • X 2 is a leaving group and is chlorine, bromine or iodine.
  • Other various substituents and groups are as defined in the specification.
  • the active substance of the present invention (the compound of the formula I, or a salt thereof, or an optical isomer thereof, or a racemate thereof, or a solvate thereof, or a precursor thereof) can be prepared into an agricultural preparation by a conventional method, for example, Solutions, emulsions, suspensions, powders, foams, pastes, granules, aerosols, natural and synthetic materials impregnated with active substances, microcapsules in polymers, bags for seeds Coating agent.
  • compositions can be produced by known methods, for example, by mixing the active compound with an extender.
  • the extender is a liquid or liquefied gas or a solid diluent or carrier, and optionally a surfactant, that is, an emulsifier and/or a dispersant and/or a foam former.
  • a surfactant that is, an emulsifier and/or a dispersant and/or a foam former.
  • an organic solvent can also be used as an auxiliary.
  • a liquid solvent When a liquid solvent is used as the diluent or carrier, it is basically suitable, such as: aromatic hydrocarbons such as xylene, toluene or alkylnaphthalene; chlorinated aromatic or chlorinated aliphatic hydrocarbons such as chlorobenzene, vinyl chloride Or methylene chloride; aliphatic hydrocarbons such as cyclohexane or paraffin, such as mineral oil fractions; alcohols such as ethanol or ethylene glycol and their ethers and lipids; ketones such as acetone, methyl ethyl ketone, methyl isobutyl Ketone or cyclohexanone; or less common polar solvents such as dimethylformamide and dimethyl sulfoxide, and water.
  • aromatic hydrocarbons such as xylene, toluene or alkylnaphthalene
  • chlorinated aromatic or chlorinated aliphatic hydrocarbons such as chlorobenzene, vinyl chloride Or
  • a diluent or carrier for a liquefied gas it is meant a liquid which will become a gas at normal temperature and pressure, such as an aerosol propellant such as a halogenated hydrocarbon and butane, propane, nitrogen and carbon dioxide.
  • the solid support can be ground natural minerals such as kaolin, clay, talc, quartz, activated clay, montmorillonite, or diatomaceous earth, and ground synthetic minerals such as highly dispersed silicic acid, alumina and silicates.
  • Solid supports for granules are ground and graded natural zircons such as calcite, marble, pumice, sepiolite and dolomite, as well as inorganic and organic coarse powder synthetic particles, and organic materials such as sawdust, coconut shell, Corn cobs and granules of tobacco stems.
  • Emulsified columns of nonionic and anionic agents can be used as emulsifiers and/or foam formers.
  • polyoxyethylene-fatty acid esters polyoxyethylene-fatty alcohol ethers, such as alkaryl polyglycol ethers, alkyl sulfonates, alkyl sulfates, aryl sulfonates, and white Protein hydrolysate.
  • Dispersing agents include, for example, lignin sulfite waste liquid and methyl cellulose.
  • Binders such as carboxymethylcellulose and natural and synthetic polymers in the form of powders, granules or emulsions such as acacia, polyvinyl alcohol and polyvinyl acetate may be employed in the formulation.
  • Coloring agents such as inorganic dyes such as iron oxide, oxidized diamonds and Prussian blue; organic dyes such as organic dyes such as azo dyes or metal phthalocyanine dyes; and trace nutrients such as iron, lanthanum, boron, copper may be used. , cobalt, aluminum and zinc salts, etc.
  • the "agronomic preparation” is generally an agricultural plant growth regulator comprising a compound of the formula I or a salt thereof, an optical isomer, a racemate, a solvate or a precursor thereof for enhancing plant stress resistance.
  • An active ingredient such as drought resistance
  • an agriculturally acceptable carrier for enhancing plant stress resistance.
  • the "agriculturally acceptable carrier” is an agrochemical for the delivery of a compound of the formula I according to the invention, or a salt, optical isomer, racemate, solvate or precursor thereof, to a plant.
  • the carrier can be a liquid or a solid.
  • the agriculturally acceptable carrier suitable for use in the present invention is selected from the group consisting of water, buffer, DMSO, a surfactant such as Tween-20, or a combination thereof. Any agriculturally acceptable carrier known to those skilled in the art can be used in the present invention.
  • the agricultural preparation of the present invention may be combined with other drought-tolerant agents in their commercial preparations or in the preparation forms prepared from these preparations.
  • These other drought-resistant agents include, but are not limited to, drought-resistant seed coating agents, drought-resistant agents.
  • the agricultural preparations of the invention may also be formulated as a mixture with the synergist in their commercial preparations or in the dosage forms prepared from these preparations, which are compounds which enhance the action of the active compound, due to the active compound It is active in itself, and it is not necessary to add a synergist.
  • the dosage form of the agricultural preparation of the present invention may be various, as long as the active ingredient can be effectively brought into the dosage form of the plant, and from the standpoint of ease of preparation and application, the preferred agricultural preparation is a spray. Agent or solution preparation.
  • the agricultural preparation of the present invention usually contains 0.0001 to 99% by weight based on the total weight of the agricultural preparation. Preferably 0.1 to 90% by weight of a compound of the invention.
  • concentration of the compound of the present invention in a commercial preparation or a used dosage form can be varied within a wide range.
  • the concentration of the compound of the invention in a commercial formulation or in a dosage form may range from 0.0000001 to 100% (g/v), preferably between 0.0001 and 1% (g/v).
  • the present invention provides a method for enhancing plant stress resistance (such as drought resistance, cold tolerance), comprising the steps of: administering to a plant a compound of formula I or a salt thereof, an optical isomer thereof, a racemate, a solvate or a former Body, or its corresponding agricultural preparation.
  • plant stress resistance such as drought resistance, cold tolerance
  • the application may be carried out by various methods known, for example, by spraying, spraying, dusting or spreading the compound or an agricultural preparation containing the compound on the plant leaf, the propagation material, or otherwise contacting the plant with the compound or containing the compound.
  • An agricultural preparation of a compound, if seeded, is coated, wrapped or otherwise treated.
  • Another method of directly treating plants or seeds prior to planting can also introduce the agricultural preparation of the present invention into soil or other medium to be seeded.
  • a carrier can also be used, which can be a solid, liquid state as described above.
  • the compound or an agricultural formulation containing the compound can also be delivered to the plant by spraying (e.g., by aircraft spraying) or irrigation.
  • a class of ABA substitutes (compounds of the invention) having high abscisic acid (ABA) activity was first developed.
  • the compounds of the present invention are capable of significantly enhancing a variety of stress resistance of plants (e.g., drought resistance, cold tolerance, etc.).
  • the compound of the present invention is simple in preparation and has excellent environmental friendliness, and therefore has broad application prospects. The present invention has been completed on this basis.
  • the compounds of the present invention can bind to a plurality of different PYL receptors, and the activity in vitro is significantly better than Abscisic Acid (ABA). It can significantly enhance the stress resistance of various plants.
  • Abscisic Acid ABA
  • the present invention is the first to synthesize a series of highly active alternative compounds of natural abscisic acid (ABA).
  • ABA abscisic acid
  • the compounds of the present invention are capable of significantly enhancing a variety of plant resistance (e.g., drought and cold tolerance). Further, the compounds of the present invention which are optical isomers or racemates have high activities.
  • the compound of the present invention promotes the interaction of a plurality of PYR/PYL receptor proteins with the PP2C protein phosphatase HAB1.
  • the model plants used in the experiments were all conventional or commercially available varieties, of which Arabidopsis thaliana includes: Columbia (Col-0) ecotype, based on the Col-0 ecotype-based ABA synthetic mutant aba2-1 and A PYL receptor triple deletion mutant based on the Col-0 ecotype (pyr1; pyl1; pyl4).
  • Soybean varieties are commercially available as Shaanxi 125 (0224, 0304, 0706, 0708, 0713, 0715 and 1028c) and Xiaxia 10 (0428, 1022B and NC0F4).
  • the cotton varieties are commercially available conventional upland cotton R15, and the wheat variety is the city. Selling Xi Nong 979, the corn variety is commercially available No. 13 on the market.
  • the growth temperature of Arabidopsis thaliana is 22 °C.
  • the photoperiod of plants grown on plant growth medium is long daylight (24 hours light), and the photoperiod of plants growing in soil (such as foliar transpiration experiment and soil drought experiment)
  • the light intensity is 75 ⁇ mol ⁇ m –2 ⁇ s –1 .
  • the plant growth medium used in the experiments was 1/2 MS (Murashige and Skoog) solid medium containing 1% (w/v) sucrose and 0.6% (w/v) agar (purchased from Phyto Technology) unless otherwise specified. Labotories company).
  • Arabidopsis gene PYL1 (amino acid sequence 36-211), PYL2 (amino acid sequence 14-188) and Arabidopsis gene HAB1 (amino acid sequence 172-511) carrying the Biotin tag sequence with the 6 ⁇ His and SUMO double-tag sequences
  • Arabidopsis gene HAB1 (amino acid sequence 172-511) carrying the Biotin tag sequence with the 6 ⁇ His and SUMO double-tag sequences
  • a gate-latch-lock mechanism for hormone signalling by abscisic acid receptors (Nature, Vol 462, 2009), PYR1 and PYL7 with 6 ⁇ His and SUMO double-tag sequences (full gene coding sequence).
  • the construction method of soybean GmPYL6 and rice OsPYL2 recombinant plasmid is the same as that of Arabidopsis thaliana PYL2.
  • the recombinant plasmid of PYR1/PYL1/PYL2/PYL7/GmPYL6/OsPYL2 carrying the 6 ⁇ His and SUMO double-tag sequences was induced with 100 ⁇ M IPTG overnight, while the HAB1 recombinant plasmid carrying the Biotin tag sequence was simultaneously induced with 100 ⁇ M IPTG and 40 ⁇ M biotin.
  • the bacterial cells after 16 hours of induction were centrifuged in a low-speed large-capacity centrifuge to collect the cells, and centrifuged at 4000 rpm for 20 min at 4 °C.
  • Each 2 L of the bacterial solution was resuspended in 50 ml of extraction buffer (containing 20 mM Tris, pH 8.0, 200 mM NaCl, and 10% (v/v) glycerin), and then crushed 3-5 times at a pressure of 1000 Pa at 4 °C.
  • the disrupted cells were subjected to ultracentrifugation, centrifuged at 16000 rpm for 30 min, and repeated twice, and the supernatant was collected through an affinity chromatography column.
  • the AlphaScreen kit (purchased from Perkin Elmer) was used as follows: 150 ⁇ l of the experimental system containing 1:10 diluted 10 ⁇ buffer (50 mM MOPS, pH 7.4, 50 mM NaF, 50 mM CHAPS, 0.1 mg/ml bovine serum albumin), each 100nM HAB1 with Biotin tag sequence and PYR1/PYL1/PYL2/PYL7/GmPYL6/OsPYL2 protein with 6 ⁇ His and SUMO double-tag sequences, corresponding concentrations of (+)-ABA/0224/0304/0706/0708/0713/ 0715/0428/NC0F4, 5 ⁇ g/ml donor beads and acceptor beads (purchased from Perkin Elmer), incubated at room temperature for 1.5 hours in the dark, placed on Envision Plate Reader (purchased from Perkin Elmer) In the ), the reading is performed according to the set AlphaScreen program.
  • 10 ⁇ buffer 50 mM MOPS, pH 7.4, 50
  • the reaction system contains 50 mM imidazole, pH 7.2, 5 mM MgCl 2 , 0.1% ⁇ -mercaptoethanol, 0.5 ⁇ g ⁇ ml -1 BSA, 100 nM HAB1 protein with Biotin tag sequence, 500 nM PYL2 with 6 ⁇ His-SUMO double tag sequence.
  • the receptor protein and the corresponding concentration of (+)-ABA/0224/0304/0706/0708/0713/0715/1028c/0428/1022B/NC0F4 were incubated for 30 minutes at room temperature, followed by the addition of a phosphorylated polypeptide containing 11 amino acids. The reaction was continued for 30 minutes.
  • the phosphorylated polypeptide was amino acids 170-180 of the SnRK2.6 protein kinase.
  • the phosphorylated serine at position 175 (sequence is HSQPKpSTVGTP, purchased from Kingsley) is a known target for dephosphorylation of HAB1.
  • Site. After 30 minutes, a chromogenic reagent (purchased from BioVision) was added, and the absorbance at a wavelength of 650 nm was read with a microplate reader (purchased from Molecular Device).
  • RNA extraction was carried out by a conventional method, followed by reverse transcription and fluorescent quantitative PCR. Three biological replicates were taken for each treatment and two experimental replicates were performed, and the ACT7 gene was used as an internal reference.
  • the Arabidopsis PYL2 and HAB1 proteins after excision and label removal were mixed with (+)-ABA or Compound 0428 in a molar ratio of 1:1:5, and concentrated to 6 mg/ml for dot crystals.
  • the spot crystal was carried out by the hanging drop method; the well buffer for crystallization contained 0.2 M sodium di-todium tartrate and 20% PEG 3350.
  • the crystal can be seen after one day and can grow to 100-120 ⁇ m in about 3-4 days.
  • the crystal is X-ray diffracted and the diffraction data is collected, and the complex is analyzed according to the relevant PYR/PYL receptor structure model. structure.
  • the compounds 0428 and 0224 of the present invention are exemplified. Seeds of Arabidopsis Col-0 ecotype and PYL receptor triple deletion mutant (pyr1; pyl1; pyl4) were sterilized with NaClO and placed at 4 °C for 3 days, then seeded at 1 ⁇ M (+)-ABA/0224. Compound or 0.05% DMSO (control) on 1/2 MS solid medium, or 1/2 MS solid medium containing 2 ⁇ M (+)-ABA/inventive compound 0428 or 0.05% DMSO (control). Two lines were sown simultaneously on each 6 cm diameter medium, and each line was sown with 15-20 seeds, with 4 replicates for each compound.
  • the medium was placed in a long-day culture at 22 ° C, and the seeds germinated on a solid medium containing 1 ⁇ M of (+)-ABA/0224 compound were photographed after 6 days of sowing, on a solid medium containing 2 ⁇ M of (+)-ABA/0428 compound. The germinated seeds were photographed 9 days after sowing.
  • the Arabidopsis thaliana leaf transpiration experiment used the ABA synthetic mutant aba2-1. Under environmental stress conditions, the endogenous ABA content in the mutant did not increase, only 1/40 of the wild-type Arabidopsis Col-0 under the same conditions, so the use of this mutant can exclude endogenous ABA for transpiration.
  • the impact of the experiment Plants sprayed for three weeks after continuous watering were sprayed with 0.05% Tween-20 and corresponding concentrations of (+)-ABA/0224/0304/0706/0715/0428 or 0.05% Tween-20 and 0.05% DMSO (control), using The amount is 1.2 ml / pot.
  • the foliar transpiration experiment of soybean was carried out at 26 ° C long day, and after spraying for 14 days, 0.1% tween-20 and 20 ⁇ M (+)-ABA/0224/0304 compound or 0.1% Tween-20 and 0.05% DMSO (control) were sprayed.
  • the solution was used in an amount of 4 ml/pot; the leaf transpiration experiment of cotton was sprayed with 0.1% tween-20 and 20 ⁇ M (+)-ABA/0224 compounds or 0.1% Tween-20 and 0.05% DMSO, respectively, after 25 days of sowing.
  • control solution was used in an amount of 4 ml/pot; the wheat leaf transpiration experiment was sprayed with 0.1% tween-20 and 100 ⁇ M (+)-ABA/0224 compound or 0.1% Tween-20 and 0.05, respectively, after 18 days of sowing.
  • a solution of % DMSO (control) was used in an amount of 6 ml per pot. All plants were imaged using a FLIR A655sc thermal imaging camera at the same time period before and after spraying.
  • the Arabidopsis Col-0 ecotype seeds were sterilized with NaClO and placed at 4 °C for 3 days and then sown on 1/2MS solid medium. After 6 days of growth, seedlings with good growth and uniform size were selected and transplanted into 8 ⁇ 7 filled with soil. ⁇ 6cm 3 in a flower pot. Each pot was filled with the same weight of soil and transferred to the same number of plants (six strains) to reduce the experimental error. All the pots were placed in a short daylight culture at 22 ° C. After two weeks, the watering was stopped for drought treatment.
  • a solution containing 0.05% Tween-20 and 5 ⁇ M of 0224/0706/0715/0428 or 0.05% Tween-20 and 0.05% DMSO (control) was sprayed onto the circumferential leaf surface at a spray rate of 2 ml solution/pot.
  • the position of the flowerpot is used to reduce the error caused by environmental factors.
  • the total solution is sprayed twice during the whole drought period, and photographed after four weeks.
  • Soybean, cotton and wheat for leaf transpiration experiments were also used for soil drought experiments, and each pot was filled with the same weight of soil to reduce experimental error. All soybean plants were cultured at 26 °C for a long period of time. After 14 days of sowing, the watering was stopped and the plants with the same growth were selected for drought treatment. At the beginning of the drought, a solution containing 0.1% Tween-20 and 20 ⁇ M (+)-ABA/0224/0304 or 0.1% Tween-20 and 0.05% DMSO (control) was sprayed onto the foliage at a spray rate of 4 ml/pot. At the same time, the position of the flowerpot was changed, and the water was rehydrated after 6 days of drought and photographed after one day of rehydration.
  • the cotton drought experiment was similar to that of soybeans. After 25 days of sowing, the watering was stopped, and the plants with the same growth were selected for drought treatment. Spraying 0.1% Tween-20 and 20 ⁇ M(+)-ABA/0224 to the foliage at the beginning of the drought Or 0.1% Tween-20 and 0.05% DMSO (control) solution, sprayed once every 3 days, sprayed 4ml / pot, while changing the position of the pot, rehydration after 6 days of drought, and before rehydration Take a photo after one day of rehydration. In the drought experiment of wheat, 16 days of consistent wheat was selected for drought treatment.
  • the soil drought test for soybean and corn for compound 0428 was similar to the soybean drought test for compound 0224, containing only one plant per pot. All soybean plants were cultured at 26 °C for a long period of time. After 3 groups of 3 leaves, the watering was stopped. The plants with the same growth were selected for drought treatment; the corn was stopped at the small bell mouth for water treatment. A solution containing 0.05% Tween-20 and 50 ⁇ M (+)-0428 or 0.05% Tween-20 and 0.05% DMSO (control) was sprayed onto the leaves on the first and second days of the drought, and the spray amount was 4 ml. / Basin, while changing the position of the pot. After 9 days of drought, the corn was rehydrated after 4 days of drought and photographed the next day.
  • the intermediate preparation method of 0708 was the same as that of 0706 except that 2-iodopropane was used instead of 1-iodoethane.
  • the intermediate preparation method of 0713 was the same as that of 0706 except that 1-iodoisobutyl was used instead of 1-iodoethane.
  • the intermediate preparation method of 0715 was the same as that of 0706 except that 1-fluoro-3-iodopropane was used instead of 1-iodoethane.
  • the reaction was allowed to stir at room temperature for 12-16 hours. At the end of the reaction, ice water was added, and ethyl acetate was extracted. Dry over anhydrous sodium sulfate and concentrate the organic phase.
  • the crude product was purified by silica gel column chromatography eluting
  • Methyl-4-nitroaniline was added to 250 ml of dry tetrahydrofuran and 1.5 equivalents of CDI were added with stirring. After the addition, the temperature was raised to reflux and stirred overnight; tetrahydrofuran was evaporated under reduced pressure, and water and dichloromethane were combined to extract mixture. The organic phases were combined, washed with a saturated aqueous solution of sodium chloride and dried over anhydrous sodium sulfate. 6-Nitro-3,4-dihydroquinazoline-2(1H)-one was obtained in a yield of 74%.
  • HAB1 protein phosphatase activity using SnRK2.6 phosphorylated polypeptide as substrate showed that compounds 0224, 0304, 0706, 0708, 0713, 0715, 1028c, 0428, 1022B and NC0F4 can promote PYL2 receptor and PP2C protein phosphoric acid.
  • the enzyme (HAB1) binds, thereby inhibiting the dephosphorylation of HAB1 to the SnRK2.6 phosphorylated polypeptide, and at a low concentration most compounds are superior or significantly superior to the same concentration of ABA.
  • PYL2 EC 50 values of the volume -HAB1 protein phosphatase complex were 1/29 and 1/15 of ABA (Fig. 2c)
  • 50 -HAB1 value PYL7 receptor protein phosphatases are complexes of the ABA-1 EC / 10 (Fig. 2d).
  • 0706,0708,0713,0715,0428,1022B NC0F4 compound and showed the same affinity for the receptor than PYL2 ABA, wherein 0706,0708,0713 and EC 0715 ABA 50 value of about 1 / 5-1 / 8 (Fig. 2e).
  • Compound 0428 has an EC 50 value of about 1/8 of ABA (Fig.
  • 1022B has an EC 50 value that is one order of magnitude lower than ABA (Fig. 2g), and the PYL2 receptor binds to HAB1 in a dose-dependent manner. effect.
  • the compound NC0F4 showed a PYR1 receptor affinity superior to ABA with an EC 50 value of about 1/11 of ABA (Fig. 2h).
  • the above plurality of compounds of the present invention are a series of PYL receptor agonists which are more efficient than existing compounds such as ABA.
  • the compounds of the present invention all exhibited significant PYR/PYL receptor affinity when the concentrations of other compounds of the present invention tested in vitro ranged from 0.01 to 100 ⁇ M.
  • the infrared camera was used to observe and record the temperature change of the leaf surface, thus reflecting the transpiration of the plant. The stronger the transpiration, the lower the leaf surface temperature.
  • the results of foliar transpiration experiments of soybean, cotton and wheat are shown in Fig. 5 and Fig. 6.
  • the foliar transpiration inhibition experiment on soybean showed that the plant surface temperature of 20 ⁇ M of compound 0224 or 0304 was still significant after two days.
  • the control group above the spraying of DMSO showed that the transpiration of soybean leaf surface was still inhibited, and the leaf surface temperature of the same concentration of ABA was the same as that of the control group (Fig. 5).
  • the foliar transpiration inhibition experiment on cotton showed that the leaf temperature of the plants with 20 ⁇ M of compound 0224 was still significantly higher than that of the control group sprayed with DMSO two days later, and the leaf temperature of the plants sprayed with the same concentration of ABA was already It was no different from the control group (Fig. 6).
  • the Arabidopsis Col-0 ecotype which was grown in soil for two weeks, stopped the feed water, and sprayed a solution containing 5 ⁇ M of compound 0224/0706/0715 or 0.05% DMSO (control) once a week during the drought, spraying 2 ml of solution.
  • the pots were sprayed twice and a 0.05% (v/v) surfactant Tween-20 was added to the solution to enhance the penetration of the spray on the leaf epidermis.
  • the control group sprayed with DMSO had dried out, while the plants sprayed with 5 ⁇ M of compound 0224/0706/0715 survived (Fig. 7).
  • Soybeans of 14 days and 25 days of cotton were separately sown, and plants of the same size were selected for soil drought experiments.
  • An aqueous solution of DMSO (control) was added with 0.1% (v/v) surfactant Tween-20 to enhance the penetration of the spray on the leaf epidermis.
  • Rehydration after 6 days of drought, spraying 20 ⁇ M of compound 0224 or 0304 compound of soybean (Fig. 8) and spraying 20 ⁇ M of compound 0224 (Fig. 9) were significantly better after rehydration than the control group sprayed with DMSO and spray Plants of the same concentration of ABA were applied.
  • Example 13 Compound 1022B can induce the expression of stress-related genes in response to ABA
  • the inventors analyzed the effect of exogenously added compound 1022B on plant gene expression.
  • the crystal structure of the PYL2-0428-HAB1 complex formed by the compound 0428 of the present invention was examined by the protein crystal analysis method described in the general method.
  • the composite crystal has a resolution of 2.4 angstroms and the control is ABA.
  • the two-dimensional structure of the two composite crystals is partially shown in Figures 11a and 11b.
  • 0428 is present in the pocket structure of PYL2.
  • the four oxygen atoms in the ABA structure can form hydrogen bonds with a plurality of water molecules with a PYL2 pocket structure and a plurality of amino acid residues of HAB1.
  • the oxygen atom and the nitrogen atom on the compound 0428 sulfonylamino group and the oxygen atom on the quinoline ring can also form a hydrogen bond.
  • the halogen substituent on the p-xylene (the fluorine atom) can also be combined with the amino acid in the PYL2 pocket structure.
  • the residue forms a hydrogen bond that further enhances the affinity of compound 0428 for the PYL2 receptor.

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Abstract

本发明公开了一种高抗逆性的植物生长调节剂及其制法和用途。具体地，本发明提供的化合物为ABA的替代物，能够显著提高植物的抗逆性，因此具有极其广泛的应用前景。

Description

高抗逆性的植物生长调节剂及其制法和用途 技术领域
本发明涉及植物学领域，具体地，涉及高抗逆性的植物生长调节剂及其制法和用途。
背景技术
脱落酸(Abscisic Acid，ABA)是平衡植物内源激素和有关生长活性物质代谢的关键因子，具有促进植物平衡吸收水、肥和协调体内代谢的能力，可有效调控植物的根/冠和营养生长与生殖生长，对提高农作物的品质、产量具有重要作用。通过施用ABA，在提高农产品品质等许多方面有着重要的生理活性作用和应用价值。此外，外源ABA能引起叶片气孔的迅速关闭，抑制蒸腾作用，可用于花的保鲜，或在作物幼苗移植栽培的运输过程中防止萎蔫。ABA还能控制花芽分化，调节花期，在花卉园艺上有很大的应用价值。
ABA可以改善作物在低温、干旱、春寒、盐渍等不良生长环境中的生长。因此，ABA具有广泛应用，可用于草坪、农田、园林，尤其是可用于西部地区等缺水地区，对于发展中国农业产业具有重大意义。
然而，天然活性的(+)-ABA不稳定且人工合成难度较大，生产成本很高。因此，ABA一直未被广泛应用于农业生产，各国科学家都在开发天然ABA的替代物。
目前虽然已经开发了一些ABA的替代物，然而这些替代物的活性尚不能令人满意，农业生产上的应用价值低。此外，一些替代物的环境友好性较差。
因此，本领域迫切需要开发环境友好的且可有效提高植物抗逆性的化合物。
发明内容
本发明的目的是提供一种环境友好的且可有效提高植物抗逆性的化合物及其制法和用途。
本发明第一方面提供了一种式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体，
Figure PCTCN2016107516-appb-000001
式中，
R1为H、卤素、C1-C3烷基、或C1-C3卤代烷基；
R2为H、卤素、C1-C3烷基、或C1-C3卤代烷基；
R3为H、卤素、C1-C3烷基、或C1-C3卤代烷基；
R4为H、卤素、C1-C3烷基、或C1-C3卤代烷基；
R5为卤素、C1-C3烷基、C1-C3卤代烷基、SF5或C3-C8环烷基；
R6为取代或未取代的C1-C7烷基、取代或未取代的C2-C7链烯基、取代或未取代的C2-C7链炔基、取代或未取代的C3-C7环烷基、或取代或未取代的-Ra-O-Rb,其中，Ra为C1-C2亚烷基而Rb为H、C1-C3烷基；并且所述的取代为具有一个或多个选自下组的取代基：卤素、-ORb、-CN、-N(Rb)2、和硝基；
R7选自下组：H、取代或未取代的C1-C6烷基、取代或未取代的C2-C6链烯基、取代或未取代的C2-C6链炔基、取代或未取代的C3-C8环烷基、取代或未取代的C5-C10杂环基、Rc-C(O)-、-ORb、-CN、和-N(Rb)2；Rc选自下组：羟基、巯基、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基；其中，所述杂环基含有1至2个选自N、O、S的杂原子，并且所述的取代为具有一个或多个选自下组的取代基：卤素、-ORb、-CN、-N(Rb)2、和硝基；
R8、R9、R10各自独立地选自下组：
(i)H；
(ii)取代或未取代的C1-C6烷基、C3-C8烷氧基、取代或未取代的C3-C8环烷基、卤素、Rc-C(O)-、-OH、-NH2；R11选自下组：羟基、巯基、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基；其中所述的取代为具有一个或多个选自下组的取代基：卤素、-ORb、-CN、-N(Rb)2、和硝基；
R11为H、C1-C3烷基、或C1-C3卤代烷基；
X为CR12、NR13、O、或S，其中R12选自下组：H、卤素、C1-C3烷基、C2-C3链烯基、C2-C3链炔基、C1-C3卤代烷基、或其组合；R13为无，或选自下组：H、卤素、C1-C3烷基、C2-C3链烯基、C2-C3链炔基、C1-C3卤代烷基、或其组合；
m＝1或2；
Figure PCTCN2016107516-appb-000002
表示单键或双键；
附件条件是，X为CR12时，
Figure PCTCN2016107516-appb-000003
为双键。
在另一优选例中，R6为C1-C7烷基、C2-C7链烯基、C2-C7链炔基、C3-C7环烷基、或-Ra-O-Rb，其中，Ra为C1-C2亚烷基而Rb为H、C1-C3烷基。
在另一优选例中，R7选自下组：取代或未取代的C1-C6烷基、取代或未取代的C2-C6链烯基、取代或未取代的C2-C6链炔基、取代或未取代的C3-C8环烷基、取代或未取代的C5-C10杂环基、Rc-C(O)-、-ORb、-CN、和-N(Rb)2；Rc选自下组：羟基、巯基、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基；其中，所述杂环基含有1至2个选自N、O、S的杂原子，并且所述的取代为具有一个或多个选自下组的取代基：卤素、-ORb、-CN、-N(Rb)2、和硝基。
在另一优选例中，当X为CR12且R12为H时，
Figure PCTCN2016107516-appb-000004
为双键。
在另一优选例中，R7为H。
在另一优选例中，当X为CR12时，R8、R9、R10不同时为H。
在另一优选例中，当X为CR12时，R8、R9、R10同时为H。
在另一优选例中，当R13为无时，
Figure PCTCN2016107516-appb-000005
为双键且m＝1。
在另一优选例中，当R13选自下组：H、卤素、C1－C3烷基、C2－C3链烯 基、C2－C3链炔基、C1-C3卤代烷基、或其组合时，
Figure PCTCN2016107516-appb-000006
为单键且m＝1或2。
在另一优选例中，当X为O或S时，
Figure PCTCN2016107516-appb-000007
为单键。
在另一优选例中，所述的卤素包括F、Cl、Br或I。
在另一优选例中，所述的卤素为F。
在另一优选例中，所述化合物具有式Ia结构：
Figure PCTCN2016107516-appb-000008
式Ia中，R1-R10、m的定义如上所述。
在另一优选例中，所述化合物具有式Ib结构：
Figure PCTCN2016107516-appb-000009
式Ib中，R1-R10、R13、m、
Figure PCTCN2016107516-appb-000010
的定义如上所述。
在另一优选例中，所述化合物具有式Ic结构：
Figure PCTCN2016107516-appb-000011
式Ic中，R1-R10、m的定义如上所述。
在另一优选例中，所述化合物具有式Id结构：
Figure PCTCN2016107516-appb-000012
式Id中，R1-R10、R12、m的定义如上所述。
在另一优选例中，R1、R2、R3、R4全为H。
在另一优选例中，R1、R2、R3、R4中有1、2、3或4个为卤素。
在另一优选例中，所述卤素包括F、Cl、Br或I。
在另一优选例中，所述卤素为F。
在另一优选例中，R1、R2、R3、R4中有4个为F。
在另一优选例中，R5为C1-C3烷基、C1-C3卤代烷基、SF5或C3-C6环烷基。
在另一优选例中，R5为甲基或环丙基。
在另一优选例中，R6为取代或未取代的C1-C5烷基、取代或未取代的C2-C5链烯基、取代或未取代的C2-C5链炔基、取代或未取代的C3-C5环烷基、或取代或未取代的-Ra-O-Rb,其中，Ra为C1-C2亚烷基而Rb为H、C1-C3烷基；并且所述的取代为具有一个或多个选自下组的取代基：卤素、-ORb、-CN、-N(Rb)2、或硝基。
在另一优选例中，R6为正丙基、乙基、异丙基、异丁基、或氟代正丙基。
在另一优选例中，R6为C3烷基、C3链烯基、或C3链炔基。
在另一优选例中，R6为正丙基。
在另一优选例中，R7选自下组：H、取代或未取代的C1-C4烷基、取代或未取代的C2-C4链烯基、取代或未取代的C2-C4炔基、取代或未取代的C3-C6环烷基、取代或未取代的C5-C8杂环基、Rc-C(O)-、-OH、-CN、和-NH2；Rc选自下组：羟基、巯基、取代或未取代的C1-C4烷基、取代或未取代的C1-C4烷氧基；其中，所述杂环基含有1至2个选自N、O、S的杂原子，并且所述的取代为具有一个或多个选自下组的取代基：卤素、-OH、-CN、-NH2、和硝基；以及
R8、R9、R10各自独立地选自下组：
(i)H；
(ii)取代或未取代的C1-C4烷基、C3-C6烷氧基、卤素、Rc-C(O)-、-OH、-NH2；Rc选自下组：羟基、巯基、取代或未取代的C1-C4烷基、取代或未取代的C1-C4烷氧基；其中所述的取代为具有一个或多个选自下组的取代基：卤素、-OH、-CN、-NH2、和硝基。
在另一优选例中，R7选自下组：H、取代或未取代的C1-C4烷基、取代或未取代的C2-C4链烯基、取代或未取代的C2-C4炔基、取代或未取代的C3-C6环烷基、取代或未取代的C5-C8杂环基、Rc-C(O)-、-OH、-CN、和-NH2；Rc选 自下组：羟基、巯基、取代或未取代的C1-C4烷基、取代或未取代的C1-C4烷氧基；其中，所述杂环基含有1个选自N、O、S的杂原子，并且所述的取代为具有一个或多个选自下组的取代基：卤素、-OH、-CN、-NH2、和硝基；以及
R8、R9、R10各自独立地选自下组：
(i)H；
(ii)取代或未取代的C1-C4烷基、C3-C6烷氧基、卤素、Rc-C(O)-、-OH、-NH2；Rc选自下组：羟基、巯基、取代或未取代的C1-C4烷基、取代或未取代的C1-C4烷氧基；其中所述的取代为具有一个或多个选自下组的取代基：卤素、-OH、-CN、-NH2、和硝基。
在另一优选例中，R7选自下组：取代或未取代的C1-C4烷基、取代或未取代的C2-C4链烯基、取代或未取代的C2-C4炔基、取代或未取代的C3-C6环烷基、取代或未取代的C5-C8杂环基、Rc-C(O)-、-OH、-CN、和-NH2；Rc选自下组：羟基、巯基、取代或未取代的C1-C4烷基、取代或未取代的C1-C4烷氧基；其中，所述杂环基含有1至2个选自N、O、S的杂原子，并且所述的取代为具有一个或多个选自下组的取代基：卤素、-OH、-CN、-NH2、和硝基；以及
R8、R9、R10各自独立地选自下组：
(i)H；
(ii)取代或未取代的C1-C4烷基、C3-C6烷氧基、卤素、Rc-C(O)-、-OH、-NH2；Rc选自下组：羟基、巯基、取代或未取代的C1-C4烷基、取代或未取代的C1-C4烷氧基；其中所述的取代为具有一个或多个选自下组的取代基：卤素、-OH、-CN、-NH2、和硝基。
在另一优选例中，R7选自下组：取代或未取代的C1-C4烷基、取代或未取代的C2-C4链烯基、取代或未取代的C2-C4炔基、取代或未取代的C3-C6环烷基、取代或未取代的C5-C8杂环基、Rc-C(O)-、-OH、-CN、和-NH2；Rc选自下组：羟基、巯基、取代或未取代的C1-C4烷基、取代或未取代的C1-C4烷氧基；其中，所述杂环基含有1个选自N、O、S的杂原子，并且所述的取代为具有一个或多个选自下组的取代基：卤素、-OH、-CN、-NH2、和硝基；以及
R8、R9、R10各自独立地选自下组：
(i)H；
(ii)取代或未取代的C1-C4烷基、C3-C6烷氧基、卤素、Rc-C(O)-、-OH、-NH2；Rc选自下组：羟基、巯基、取代或未取代的C1-C4烷基、取代或未取代的C1-C4烷氧基；其中所述的取代为具有一个或多个选自下组的取代基：卤素、-OH、-CN、-NH2、和硝基。
在另一优选例中，R7、R8、R9、R10为本申请实施例中各具体化合物中相对应的具体基团。
在另一优选例中，所述化合物选自下组：
Figure PCTCN2016107516-appb-000013
Figure PCTCN2016107516-appb-000014
Figure PCTCN2016107516-appb-000015
Figure PCTCN2016107516-appb-000016
在另一优选例中，所述化合物选自下组：
Figure PCTCN2016107516-appb-000017
本发明第二方面提供了一种本发明第一方面所述式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体的用途，用于制备农用制剂或组合物，所述农用制剂或组合物用于(i)增强植物抗逆性；(ii)制备ABA受体的激动剂；和/或(iii)制备种子萌发抑制剂。
在另一优选例中，所述的激动剂促进ABA受体PYL蛋白与PP2C蛋白磷酸酶的相互作用。
在另一优选例中，所述农用制剂或组合物用于一种或多种以下用途：
(i)促进ABA受体PYL蛋白与PP2C蛋白磷酸酶的相互作用；
(ii)减弱叶片的蒸腾作用；
(iii)抑制种子萌发。
在另一优选例中，所述抗逆性为ABA相关的非生物胁迫抗逆性。
在另一优选例中，所述抗逆性选自下组：抗旱性、耐冷性、耐盐碱、耐渗透压、耐热性、或其组合。
在另一优选例中，所述植物为含有PYR/PYL家族ABA受体的植物。
在另一优选例中，所述植物包括苔藓、蕨类、裸子植物、单子叶植物和双子叶植物。
在另一优选例中，所述的植物包括农业植物、园艺植物、林业植物。
在另一优选例中，所述的植物包括木本植物、草本植物。
在另一优选例中，所述的植物包括完整的植株、器官(如根、茎、叶、枝、花、果实、种子)、组织(如愈伤组织)、或细胞。
在另一优选例中，所述的植物选自下组：禾本科、菊科、百合科、十字花科、蔷薇科、豆科、茶科、梧桐科、松科、胡桃科、胡椒科、木兰科、杜鹃花科、猕猴桃科、葡萄科、秋海棠科、凤梨科、银杏科、八角茴香科、姜科、石榴科、夹竹桃科、小檗科、芸香科、茄科、柏科、冬青科、棕榈科植物、或其组合。
在另一优选例中，所述植物选自下组：拟南芥、烟草、棉花、生菜、水稻、小麦、玉米、花生、高粱、燕麦、黑麦、甘蔗、大豆、马铃薯、荞麦、胡椒、葡萄、梨、苹果、香蕉、人参、番茄、辣椒、茄子、花椰菜、大白菜、油菜、黄瓜、西瓜、洋葱、向日葵、百合、玫瑰、菊花、牡丹、康乃馨、樟树、梧桐、松树、或其组合。
本发明第三方面提供了一种农用制剂，所述农用制剂包括：
(i)本发明第一方面所述的式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体；和
(ii)农业上可接受的载体。
在另一优选例中，组分(i)在农用制剂中的含量为0.1-1000μM，较佳地，1-200μM，更佳地，5-100μM。
在另一优选例中，所述农用制剂中，含有0.0001-99wt％，较佳地0.1-90wt％的组分(i)，以农用制剂的总重量计。
在另一优选例中，所述农用制剂还包括额外的抗旱剂(如抗旱种衣剂、抗旱保水剂、或抗旱喷洒剂)或其他农用活性成分。
在另一优选例中，所述的农用活性成分选自下组：杀真菌剂、除草剂、杀虫剂、杀线虫剂、杀昆虫剂、植物激活剂、增效剂、植物生长调节剂、杀螨剂。
在另一优选例中，所述农用制剂还包括表面活性剂(如阳离子型、阴离子型、两性、或非离子型表面活性剂)。
在另一优选例中，所述农用制剂的剂型选自下组：溶液剂、乳剂、混悬剂、粉剂、泡沫剂、糊剂、颗粒剂、气雾剂、或其组合。
本发明第四方面提供了一种增强植物抗逆性的方法，给所述植物施用本发明第一方面所述的式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体或施用本发明第三方面所述的农用制剂。
在另一优选例中，所述施用选自下组：喷洒或灌溉。
在另一优选例中，所述施用的剂量为2-100g/公顷，较佳地，4-80g/公顷，更佳 地，6-60g/公顷。
在另一优选例中，所述施用的剂量为1-5000微克/株，较佳地，10-2500微克/株，更佳地，20-1000微克/株。
本发明第五方面提供了一种式I化合物或其盐的制法，包括步骤：
(a)在惰性溶剂中，将化合物I-A与化合物I-S2进行反应，从而形成式I化合物；
Figure PCTCN2016107516-appb-000018
上述各式中，R1、R2、R3、R4、R5、R6、R7、R8、R9、R10、R11、m、X、
Figure PCTCN2016107516-appb-000019
如本发明第一方面所定义。
在另一优选例中，所述惰性溶剂选自下组：N，N-二甲基甲酰胺(DMF)、二氯甲烷(DCM)、乙腈(ACN)、或其组合。
在另一优选例中，所述反应在缚酸剂存在下进行。
在另一优选例中，所述缚酸剂选自下组：碳酸钾(K2CO3)、三乙胺(Et3N)、吡啶(Py)、或其组合。
在另一优选例中，步骤(a)中，所述反应温度为0－150℃(或回流温度)，较佳地，10－60℃，更佳地，20－40℃。
在另一优选例中，步骤(a)中，所述反应时间为0.1-72小时，更佳地，1-24小时，更佳地，8-20小时，更佳地，4-12小时。
在另一优选例中，在式I-A中，X为O，且
Figure PCTCN2016107516-appb-000020
为单键。
在另一优选例中，所述化合物I-S2由如下方法制备：
(i)在惰性溶剂中，将化合物I-SS1与硫脲反应，从而形成化合物I-S2；
Figure PCTCN2016107516-appb-000021
上述各式中，R1、R2、R3、R4、R5如本发明第一方面所定义，X2为离去基团(如Cl、Br或I)。
在另一优选例中，所述惰性溶剂选自下组；乙醇、乙腈、四氢呋喃、或其组合。
在另一优选例中，所述反应在酸条件下进行。
在另一优选例中，所述酸选自下组：盐酸、氢溴酸、或其组合。
在另一优选例中，在步骤(i)中，所述反应温度为0－150℃(或回流温度)，较佳地，10－50℃，更佳地，15－25℃。
在另一优选例中，在步骤(i)中，所述反应时间为0.1-72小时，更佳地，1-24小时，更佳地，2-12小时。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
附图说明
图1显示多个本发明化合物，包括0224(图1a)，0304(图1b)，0706，0708，0713，0715(图1c)，1028c(图1d)，0428(1e)，1022B(1f)和NC0F4(1g)可以结合拟南芥PYL2受体-HAB1复合物，进而抑制蛋白磷酸酶HAB1的活性。且在更低浓度下，上述化合物均有抑制效果，并且大多数化合物的抑制效果优于或显著优于ABA。
图2显示了多个本发明化合物作为PYL受体激动剂存在计量依赖效应。其中0224和0304与拟南芥PYR1(图2a),PYL1(图2b),PYL2(图2c)和PYL7(图2d)受体、以及0706，0708，0713和0715四种化合物(图2e)、化合物0428(图2f)、1022B(图2g)和NC0F4(图2h)与拟南芥PYL2受体的受体激动剂的剂量响应曲线表明上述化合物可促进拟南芥蛋白磷酸酶HAB1与PYL受体的相互作用，且该相互作用存在剂量依赖效应。EC50数值显示上述化合物与相应受体的亲和性均显著高于ABA。
图3显示1μM浓度下，0224化合物和ABA对于Col-0和pyr1；pyl1；pyl4三突变体种子萌发的影响。每个培养皿的左半边播种Col-0，右半边播种pyr1；pyl1；pyl4三突变体。图片为pyr1；pyl1；pyl4三突变体种子萌发4天后(播种6天后)所获得的。DMSO处理为对照组。结果显示0224化合物可以抑制Col-0种子的萌发，但对pyr1；pyl1；pyl4三突变体种子的萌发抑制作用显著减弱，这表明0224化合物对于拟南芥种子萌发的抑制是通过ABA受体介导的而非毒性作用。
图4显示了本发明化合物0224，0304，0706，0715或0428处理显著降低了拟南芥叶面的蒸腾速率，导致叶面温度升高。其中，图4a显示5μM ABA或化合物0224处理后，叶面温度较对照组(DMSO)处理显著增加，且化合物0224的持续时间更久；图4b则显示5μM化合物0304处理后，叶面温度较对照组(DMSO)处理显著增加，而化合物0224浓度降低至2μM或1μM后，叶面温度较对照组(DMSO)处理仍显著增加，且效果逐级递减，这表明化合物0224对于叶面蒸腾作用的抑制效应存在浓度依赖效应；图4c则显示5μM化合物0706或0715处理后，叶面温度较对照组(DMSO)处理显著增加，其持续时间与0224相当；图4d显示了5μM、2μM和1μM化合物0428处理后，叶面温度较DMSO处理显著增加，且叶面温度随浓度增高而增高，表明化合物0428对于叶面蒸腾作用的抑制效应存在浓度依赖效应。
图5显示了本发明化合物0224和0304处理后可显著降低大豆叶面的蒸腾速率，导致叶面温度升高。播种14天后的大豆植株停止浇水，同时喷施本发明的化合物0224、0304或ABA，与对照组(DMSO)相比，20μM的化合物 0224或0304均可显著降低大豆叶面的蒸腾速率，且抑制效果优于同浓度ABA。
图6显示了本发明化合物0224处理显著降低了棉花叶面的蒸腾速率，导致叶面温度升高。播种25天后的棉花植株停止浇水，同时喷施本发明的化合物0224或ABA，与对照组(DMSO)相比，20μM的化合物0224均可显著降低棉花叶面的蒸腾速率，且抑制效果优于同浓度ABA。
图7显示了拟南芥的土壤干旱实验结果。拟南芥在干旱前和干旱四周后拍照，对照组(DMSO处理)的拟南芥在干旱四周后已干枯，经化合物0224，0706或0715处理的拟南芥仍正常生长。
图8显示了大豆的土壤干旱实验结果。图5中的大豆在干旱6天后复水，照片显示了复水一天后的大豆生长状况。经化合物0224或0304处理的大豆长势要显著优于对照组(DMSO)或同浓度ABA处理的大豆。
图9显示了棉花的土壤干旱实验结果。图6中的棉花在干旱6天后复水，期间每隔3天喷施一次本发明的化合物0224或ABA，照片显示了复水前和复水一天后的棉花生长状况。经化合物0224处理的棉花长势要显著优于对照组(DMSO)或同浓度ABA处理的棉花。
图10显示了用10μM本发明化合物1022B处理6小时后，野生型拟南芥体内受ABA诱导的胁迫相关基因的转录水平变化。DMSO和同浓度ABA处理分别为阴性和阳性对照组。结果显示本发明化合物1022B诱导的4个胁迫相关基因转录水平均高于ABA。
图11a和11b分别显示了ABA(a)或本发明化合物(0428)(b)与PYL2-HAB1复合体口袋结构内多个氨基酸残基相互作用的二维结构。图中显示了水分子、氮原子、氧原子和卤素原子，虚线代表氢键，其上的数字则表示两个原子/分子之间的距离(单位是埃，
Figure PCTCN2016107516-appb-000022
)。结果显示，与ABA类似，本发明的化合物0428与PYL2口袋结构内的氨基酸残基形成多个氢键，不同的是这些氢键的形成并不需要水分子介导，这有助于化合物0428与PYL2-HAB1复合体的结合更加紧密。
图12显示了拟南芥土壤干旱实验结果。短日照环境中生长3周的野生型拟南芥植物(Col-0)停止浇水并喷施5μM ABA或本发明化合物0428。图片显示的分别是首次喷施化合物当天和14天后的植物生长状况，喷施DMSO溶液的植物作为负对照。结果显示喷施化合物0428的植物长势优于对照组和喷施ABA的植物。
图13显示本发明的化合物(如化合物0428)及ABA的大豆GmPYL6和水稻OsPYL2受体激动剂剂量响应曲线。化合物0428可以促进拟南芥蛋白磷酸酶HAB1与大豆GmPYL6或水稻OsPYL2的相互作用，且该相互作用存在剂量依赖效应。
图14a和图14b分别显示了大豆和玉米的土壤干旱实验结果。选取小喇叭口期的玉米和3组3叶期的大豆植株，在开始干旱后的第一天和第二天喷施本发明的化合物(如化合物0428)，照片显示了玉米干旱处理四天和大豆干旱处理九天后的整体生长状况。实验中化合物0428的浓度为50μM。经化合物0428处理的玉米和大豆的长势要显著优于对照组。
图15显示2μM浓度下，化合物0428和ABA对于Col-0和pyr1；pyl1；pyl4三突变体种子萌发的影响。每个培养皿的左半边播种Col-0，右半边播种 pyr1；pyl1；pyl4三突变体。照片为pyr1；pyl1；pyl4三突变体种子萌发7天后(播种9天后)所拍摄的。DMSO处理为对照组。结果显示0428化合物可以抑制Col-0种子的萌发，但对pyr1；pyl1；pyl4三突变体种子的萌发抑制作用显著减弱，这表明0428化合物对于拟南芥种子萌发的抑制是通过ABA受体介导的而非毒性作用。
图16显示了本发明化合物0224处理显著降低了小麦叶面的蒸腾速率，导致叶面温度升高。播种18天后的小麦植株停止浇水，同时喷施本发明的化合物0224，与对照组(DMSO)相比，100μM的化合物0224可显著降低小麦叶面的蒸腾速率。
图17显示小麦的土壤干旱实验结果。图16中的小麦在干旱6天后拍照，照片显示经100μM化合物0224或ABA处理的小麦长势均显著优于对照组(DMSO)。
具体实施方式
本发明人经过广泛而深入的研究，首次开发了一类具有高脱落酸(ABA)活性的ABA替代物(本发明化合物)。本发明化合物能够显著增强植物的多种抗逆性(如抗旱、耐冷等)。此外，本发明化合物的制法简便，且具有优异的环境友好性和作用迅速等优点，因此，具有广泛的应用前景。在此基础上完成了本发明。
实验表明，本发明化合物可结合多个PYR/PYL受体，体外活性优于脱落酸(Abscisic Acid，ABA)，而且可显著增强各种不同植物的抗逆性。
基团定义
如本文所用，术语“取代或未取代的”指所述基团可以是未取代的，或者所述基团中的H被一个或多个(如1-10个，较佳地1-5个，更佳地1-3个，最佳地，1-2个)取代基所取代。
如本文所用，所述的“取代”或“取代的”指所述基团具有一个或多个(较佳地1-6个，更佳地1-3个)选自下组的取代基：卤素、羟基、-NH2、硝基、-CN、C1-C4烷基、C1-C4卤代烷基、C1-C4烷氧基、C3-C6环烷基、C1-C3羧基、C2-C4链烯基、C2-C4炔基。
如本文所用，术语“C1-C7烷基”是指具有1-7个碳原子的直链或支链烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C1-C7烷基。
如本文所用，术语“C1-C6烷基”是指具有1-6个碳原子的直链或支链烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C1-C6烷基。
如本文所用，术语“C1-C4烷基”是指具有1-4个碳原子的直链或支链烷基，例如甲基、乙基、丙基、异丙基、丁基、异丁基、仲丁基、叔丁基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C1-C4烷基。
如本文所用，术语“C1-C3烷基”是指具有1-3个碳原子的直链或支链烷基，例如甲基、乙基、正丙基、异丙基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C1-C3烷基。
如本文所用，术语“C1-C2亚烷基”是指具有1-2个碳原子的二价烃基，如，亚甲基、亚乙基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C1-C2亚烷基。
如本文所用，术语“C2-C7链烯基”指具有2-7个碳原子的直链或支链的烯基，例如乙烯基、烯丙基、1-丙烯基、异丙烯基、1-丁烯基、2-丁烯基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C2-C7链烯基。
如本文所用，术语“C2-C6链烯基”指具有2-6个碳原子的直链或支链的烯基，例如乙烯基、烯丙基、1-丙烯基、异丙烯基、1-丁烯基、2-丁烯基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C2-C6链烯基。
如本文所用，术语“C2-C3链烯基”指具有2-3个碳原子的直链或支链的烯基，例如乙烯基、烯丙基、1-丙烯基、异丙烯基或类似基团。在未特别指出的时候，该术语包括取代或未取代的C2-C3链烯基。
如本文所用，术语“C2-C7链炔基”是指具有2-7个碳原子的直链或支链的炔基，例如乙炔基、丙炔基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C2-C7链炔基。
如本文所用，术语“C2-C6链炔基”是指具有2-6个碳原子的直链或支链的炔基，例如乙炔基、丙炔基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C2-C6链炔基。
如本文所用，术语“C2-C3链炔基”是指具有2-3个碳原子的直链或支链的炔基，例如乙炔基、丙炔基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C2-C3链炔基。
如本文所用，术语“C3-C8环烷基”指具有3-8个碳原子的环状烷基，例如环丙基、环丁基、环戊基、环己基、环庚基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C3-C8环烷基。
如本文所用，术语“C3-C7环烷基”指具有3-7个碳原子的环状烷基，例如环丙基、环丁基、环戊基、环己基、环庚基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C3-C7环烷基。
如本文所用，术语“C3-C6环烷基”指具有3-6个碳原子的环状烷基，例如环丙基、环丁基、环戊基、环己基、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C3-C6环烷基。如本文所用，术语“C5-C10杂环基”指饱和的、部分饱和的或不饱和的基团(但不是芳香性的)，具有单环或稠环(包括桥环体系和螺环体系，具有5至10个碳原子和1至2个选自氮、硫或氧的杂原子)，在稠环体系中，一个或多个环可以是环烷基、芳基或杂芳基，只要连接点通过非芳香性环。该术语包括取代或未取代的杂环基。
如本文所用，术语“C5-C8杂环基”指饱和的、部分饱和的或不饱和的基团(但不是芳香性的)，具有单环或稠环(包括桥环体系和螺环体系，具有5至8个碳原子和1至2个选自氮、硫或氧的杂原子)，在稠环体系中，一个或多个环可以是环烷基、芳基或杂芳基，只要连接点通过非芳香性环。该术语包括取代或未取代的杂环基。
如本文所用，术语“C1-C3卤代烷基”是指氢被1个或1个以上的卤素取代的具有1-3个碳原子的直链或支链烷基，例如，卤代甲基、卤代乙基、卤代丙基、卤代异丙基、或类似基团。在未特别指出的时候，该术语包括取代或未 取代的C1-C3卤代烷基。
如本文所用，术语“C1-C6烷氧基”是指具有“(C1-C6烷基)-O-结构的基团，例如，CH3-O-、C2H5-O-、C3H7-O-、(CH3)2CH-O-、nC4H9-O-、tC4H9-O-、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C1-C6烷氧基。
如本文所用，术语“C1-C4烷氧基”是指具有“(C1-C4烷基)-O-结构的基团，例如，CH3-O-、C2H5-O-、C3H7-O-、(CH3)2CH-O-、nC4H9-O-、tC4H9-O-、或类似基团。在未特别指出的时候，该术语包括取代或未取代的C1-C4烷氧基。
如本文所用，术语“卤素”是指氟、氯、溴、或碘，优选氟和氯，最优选氟。
如本文所用，术语“卤代的”指被相同或不同的一个或多个上述卤原子取代的基团，可以部分卤代或全部卤代，例如三氟甲基、五氟乙基、七氟异丙基、或类似基团。
本发明的化合物可以含有一个或多个不对称中心，并因此以消旋体、外消旋混合物、单一对映体、非对映异构体化合物和单一非对映体的形式出现。可以存在的不对称中心，取决于分子上各种取代基的性质。每个这种不对称中心将独立地产生两个旋光异构体，并且所有可能的旋光异构体和非对映体混合物和纯或部分纯的化合物包括在本发明的范围之内。本发明包括化合物的所有异构形式。
本发明化合物及其制备方法
如本文所用，术语“本发明化合物”、“本发明ABA替代物”、“式I化合物”可互换使用，均指具有式I所示结构的化合物。此外，所述术语还包括式I化合物的盐、光学异构体、外消旋体、溶剂化物(如水合物)、和/或前体，
Figure PCTCN2016107516-appb-000023
其中，R1-R10、R11、m、X、
Figure PCTCN2016107516-appb-000024
的定义如前所述。
在另一优选例中，所述化合物具有式Ia结构：
Figure PCTCN2016107516-appb-000025
式Ia中，R1-R10、m的定义如上所述。
在另一优选例中，所述化合物具有式Ib结构：
Figure PCTCN2016107516-appb-000026
式Ib中，R1-R10、R13、m、
Figure PCTCN2016107516-appb-000027
的定义如上所述。
在另一优选例中，所述化合物具有式Ic结构：
Figure PCTCN2016107516-appb-000028
式Ic中，R1-R10、m的定义如上所述。
在另一优选例中，所述化合物具有式Id结构：
Figure PCTCN2016107516-appb-000029
式Id中，R1-R10、R12、m的定义如上所述。
下面更具体地描述本发明式I化合物的制备方法，但这些具体方法不对本发明构成任何限制。本发明化合物还可以任选将在本说明书中描述的或本领域已知的各种合成方法组合起来而方便的制得，这样的组合可由本发明所属领域的技术人员容易的进行。通常，在本发明的制备方法中，各反应大多在惰性溶剂中，在0℃至150℃(或回流温度)(较佳地，10-60℃，或20－40℃)下，反应一段时间(如0.1-72小时，较佳地2－20小时)。
如本文所用，室温是指4-35℃，优选20-30℃。
较佳地，本发明式I化合物可以通过以下方案及实施例中所述的示例性方法以及本领域技术人员所用的相关公开文献操作完成。
典型地，本发明的式Ia化合物的制备方法可包括(但不限于)如下流程。
方案I(以X＝O，R6＝丙基，R7＝甲基，R8＝H，R9＝H，R10＝H，m＝1 为例)
(1)制备1-丙基-4-甲基-6-氨基-1,4-二氢苯并噁唑-2-酮：
Figure PCTCN2016107516-appb-000030
在步骤1中，在惰性溶剂(如四氢呋喃)中，首先将式I-1化合物与N,N'-羰基二咪唑(CDI)在一定温度(如20－40℃)下进行反应一段时间，形成式I-2化 合物。
步骤2：在碱(如氢化钠)存在下，在惰性溶剂(如N，N-二甲基甲酰胺)中，式I-2化合物与碘丙烷在一定温度(如20－40℃)下进行反应一段时间，形成式I-3化合物。
步骤3：在酸(如硫酸)存在下，式I-3化合物与硝酸钾在一定温度(如0－10℃)下进行反应一段时间，形成式I-4化合物。
步骤4：在惰性溶剂(如甲醇)中，以钯碳作催化剂，在一定温度(如20－40℃)下，将式I-4化合物进行还原反应，形成式I-5化合物。
(2)制备4-甲基卤代苄磺酰氯：
Figure PCTCN2016107516-appb-000031
步骤5：在惰性溶剂(如乙醇、乙腈)中，式I-SS1化合物(如2，3，5，6-四氟-4-甲基苄溴或2，3，5，6-四氟-4-甲基苄氯)与硫脲反应，形成反应产物；然后，在酸(如浓盐酸)存在下，在惰性溶剂(如乙腈)中，将所述反应产物和亚氯酸钠在一定温度(如15-25℃)下进行反应一段时间，形成式I-S2化合物。
(3)制备式Ia化合物
Figure PCTCN2016107516-appb-000032
步骤6：在惰性溶剂(如DMF)中，在缚酸剂(如碳酸钾)存在下，式I-5化合物与式I-S2化合物在一定温度(如20－50℃)下进行反应一段时间，从而得到式Ia化合物。
方案I中，X2为离去基团，为氯、溴、碘。其它各取代基和基团如说明书中所定义。
农用制剂
可将本发明的活性物质(式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体)以常规的方法制备成农用制剂，例如溶液剂、乳剂、混悬剂、粉剂、泡沫剂、糊剂、颗粒剂、气雾剂、用活性物质浸渍的天然的和合成的材料、在多聚物中的微胶囊、用于种子的包衣剂。
这些制剂可用已知的方法生产，例如，将活性化合物与扩充剂混合，这些 扩充剂就是液体的或液化气的或固体的稀释剂或载体，并可任意选用表面活性剂即乳化剂和/或分散剂和/或泡沫形成剂。例如在用水作扩充剂时，有机溶剂也可用作助剂。
用液体溶剂作稀释剂或载体时，基本上是合适的，如：芳香烃类，例如二甲苯，甲苯或烷基萘；氯化的芳香或氯化的脂肪烃类，例如氯苯，氯乙烯或二氯甲烷；脂肪烃类，例如环己烷或石蜡，例如矿物油馏分；醇类，例如乙醇或乙二醇以及它们的醚和脂类；酮类，例如丙酮，甲乙酮，甲基异丁基酮或环已酮；或不常用的极性溶剂，例如二甲基甲酰胺和二甲基亚砜，以及水。
就液化气的稀释剂或载体说，指的是在常温常压下将成为气体的液体，例如气溶胶推进剂，如卤化的烃类以及丁烷，丙烷，氮气和二氧化碳。
固体载体可用研磨的天然矿物质，例如高岭土，粘土，滑石，石英，活性白土，蒙脱土，或硅藻土，和研磨合成的矿物质，例如高度分散的硅酸，氧化铝和硅酸盐。供颗粒用的固体载体是碾碎的和分级的天然锆石，例如方解石，大理石，浮石，海泡石和白云石，以及无机和有机粗粉合成的颗粒，和有机材料例如锯木屑，椰子壳，玉米棒子和烟草梗的颗粒等。
非离子的和阴离子的乳化列可用作乳化剂和/或泡沫形成剂。例如聚氧乙烯-脂肪酸酯类，聚氧乙烯-脂肪醇醚类，例如烷芳基聚乙二醇醚类，烷基磺酸酯类，烷基硫酸酯类，芳基磺酸酯类以及白蛋白水解产物。分散剂包括，例如木质素亚硫酸盐废液和甲基纤维素。
在制剂中可以用粘合剂，例如羧甲基纤维素和以粉末，颗粒或乳液形式的天然和合成的多聚物，例如阿拉伯胶，聚乙烯基醇和聚乙烯醋酸酯。
可以用着色剂例如无机染料，如氧化铁，氧化钻和普鲁士蓝；有机染料，如有机染料，如偶氮染料或金属钛菁染料；和用痕量营养剂，如铁，猛，硼，铜，钴，铝和锌的盐等。
在本发明中，所述“农用制剂”通常是农用植物生长调节剂，其含有式I化合物或其盐、其光学异构体、外消旋体、溶剂化物或前体作为增强植物抗逆性(如抗旱性)的活性成分；以及农业上可接受的载体。
如本文所用，所述“农业上可接受的载体”是用于将本发明的式I化合物或其盐、光学异构体、外消旋体、溶剂化物或前体传送给植物的农药学上可接受的溶剂、悬浮剂或赋形剂。载体可以是液体或固体。适用于本发明的农业上可接受的载体选自下组：水、缓冲液、DMSO、表面活性剂如Tween-20、或其组合。任何本领域技术人员已知的农业上可接受的载体均可用于本发明中。
本发明的农用制剂可与其他抗旱剂制成一种混合物存在于它们的商品制剂中或从这些制剂制备的使用剂型中，这些其他的抗旱剂包括(并不限于)：抗旱种衣剂、抗旱保水剂、或抗旱喷洒剂等。
此外，本发明的农用制剂也可与增效剂制成一种混合物存在于它们的商品制剂中或从这些制剂制备的使用剂型中，这些增效剂是提高活性化合物作用的化合物，由于活性化合物本身有活性，也可不必加增效剂。
本发明所述的农用制剂的剂型可以是多种多样的，只要能够使活性成分有效地到达植物体内的剂型都是可以的，从易于制备和施用的立场看，优选的农用制剂是一种喷雾剂或溶液制剂。
本发明所述的农用制剂通常含有占所述农用制剂总重量的0.0001-99wt％， 较佳地0.1-90wt％的本发明化合物。商品制剂或使用剂型中的本发明化合物的浓度可在广阔的范围内变动。商品制剂或使用剂型中的本发明化合物的浓度可从0.0000001-100％(g/v)，最好在0.0001与1％(g/v)之间。
增强植物抗逆性的方法
本发明提供了一种增强植物抗逆性(如抗旱性、耐冷性)的方法，包括步骤：给植物施用式I化合物或其盐、其光学异构体、外消旋体、溶剂化物或前体、或其相应的的农用制剂。
施用可采用已知的各种方法，例如，通过在植物叶片、繁殖材料上喷洒、喷雾、喷粉或播撒该化合物或含有该化合物的农用制剂，或以其他方式使植物接触该化合物或含有该化合物的农用制剂，如果是种子，则通过涂布、包裹或以其他方式处理种子。另一种在种植前直接处理植物或种子的方法，还可将本发明的农用制剂引入土壤或其他待播种种子的培养基。在一些实施方案中，还可以使用载体，所述载体可以是如上所述的固态、液态。
在一优选实施方式中，还可以通过喷洒(如飞机喷洒)或灌溉将该化合物或含有该化合物的农用制剂递送给所述植物。
本发明的主要优点包括：
首次开发了一类具有高脱落酸(ABA)活性的ABA替代物(本发明化合物)。本发明化合物能够显著增强植物的多种抗逆性(如抗旱、耐冷等)。此外，本发明化合物制法简便，且具有优异的环境友好性，因此，具有广泛的应用前景。在此基础上完成了本发明。
实验表明，本发明化合物(如化合物0224、0304，0706，0708，0713，0715，0428，1022B等)可结合于多个不同PYL受体，体外活性显著优于脱落酸(Abscisic Acid，ABA)，可显著增强各种不同植物的抗逆性。
(1)本发明首次合成了一系列的天然脱落酸(Abscisic Acid，ABA)的高活性的替代化合物。本发明的化合物能够显著增强植物多种抗逆性(如抗旱、耐冷)。并且，光学异构体或外消旋体的本发明化合物均具有高活性。
(2)本发明化合物的活性显著优于脱落酸(Abscisic Acid，ABA)以及现有的ABA类似物。
(3)本发明的化合物对于多个PYR/PYL受体蛋白与PP2C蛋白磷酸酶HAB1的相互作用均有促进作用。
(4)本发明化合物的合成方法简单、成本低廉。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件，例如Sambrook等人，分子克隆：实验室手册(New York:Cold Spring Harbor Laboratory Press,1989)中所述的条件，或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数是重量百分比和重量份数。
除非有特别说明，否则本发明实施例中所用的材料和试剂均为市售产品。
材料和通用方法
材料
实验中使用的模式植物均为常规的或市售的品种，其中拟南芥(Arabidopsis thaliana)包括：哥伦比亚(Col-0)生态型，基于Col-0生态型的ABA合成突变体aba2-1和基于Col-0生态型的PYL受体三缺失突变体(pyr1；pyl1；pyl4)。大豆品种为市售陕豆125(0224、0304，0706，0708，0713，0715和1028c)和邯夏10号(0428、1022B和NC0F4)，棉花品种为市售常规陆地棉R15，小麦品种为市售西农979，玉米品种为市售掖单13号。
本发明所述的各化合物(如0224、0304，0706，0708，0713，0715，1028c，0428，1022B和NC0F4等)见各实施例。
植物生长
拟南芥生长温度为22℃，生长在植物生长培养基上的植物的光周期为长日照(24小时光照)，生长在土壤中的植物(如叶面蒸腾实验和土壤干旱实验)的光周期为短日照(8小时光照/16小时黑暗)，光强为75μmol·m–2·s–1。
大豆和棉花生长温度为26℃，光周期为14小时光照/10小时黑暗，玉米的生长温度为27℃，光周期为11小时光照/13小时黑暗，小麦的生长温度为27℃，光周期为11小时光照/13小时黑暗，光强均为400μmol·m–2·s–1。
如无特殊指明，实验中使用的植物生长培养基均为含1％(w/v)蔗糖和0.6％(w/v)琼脂的1/2MS(Murashige and Skoog)固体培养基(购自Phyto Technology Labotories公司)。
蛋白表达纯化
带6×His和SUMO双标签序列的拟南芥基因PYL1(氨基酸序列36-211)，PYL2(氨基酸序列14-188)和带Biotin标签序列的拟南芥基因HAB1(氨基酸序列172-511)的重组质粒的构建方法详见“A gate-latch-lock mechanism for hormone signalling by abscisic acid receptors”(Nature,Vol 462,2009)，带6×His和SUMO双标签序列的PYR1和PYL7(全基因编码序列)以及大豆GmPYL6和水稻OsPYL2重组质粒的构建方法与拟南芥PYL2相同。
将上述重组质粒转入感受态细胞大肠杆菌BL21(DE3)(购自NEB公司)，接种到含有Amp抗性的200ml LB液体培养基(购自OXOID公司)，37℃200rpm培养过夜；按1:50-1:100接种至2L含有Amp抗性的LB液体培养基中进行扩大培养，37℃200rpm培养3-4小时，16℃低温培养至OD600＝0.8-1.0左右。带6×His和SUMO双标签序列的PYR1/PYL1/PYL2/PYL7/GmPYL6/OsPYL2的重组质粒用100μM IPTG诱导过夜，而带Biotin标签序列的HAB1重组质粒要用100μM IPTG和40μM biotin同时进行诱导。
将诱导16小时后的菌液在低速大容量离心机中离心收集菌体，4℃以4000rpm转速离心20min。每2L菌液用50ml提取缓冲液(含20mM Tris，pH 8.0，200mM NaCl和10％(v/v)甘油)重悬菌体，然后在4℃进行1000Pa压力破碎3-5次。破碎后的细胞进行超速离心，16000rpm离心30min，重复2次，收集上清液过亲和层析柱。
对于带6×His和SUMO双标签序列的PYR/PYL蛋白，选择50ml亲和层析Ni柱(50ml Ni-NTA column，购自GE公司)；先用10％缓冲液B(含20mM Tris，pH 8.0，200mM NaCl，500mM imidazole和10％甘油)平衡600ml，再用200 ml 50％缓冲液B洗脱，最后用100ml 100％缓冲液B洗脱；用于晶体解析的蛋白与ulp1酶以1000：1的摩尔比混合进行酶切透析过夜；酶切后的蛋白再过一次亲和层析Ni柱；收集液用洗脱溶液(含25mM Tris，pH 8.0，200mM ammonium acetate，1mM dithiotreitol和1mM EDTA)过HiLoad 26/60Superdex200凝胶过滤柱(购自GE公司)进一步分离纯化蛋白。
对于带Biotin标签序列的HAB1蛋白，过50ml MBP亲和层析柱(购自GE公司)；先用10％缓冲液C(含20mM Tris，pH 8.0，200mM NaCl，10mM Maltose和10％甘油)平衡600ml，再用200ml 50％缓冲液C洗脱，最后用100ml 100％缓冲液C洗脱；收集液用洗脱溶液(含20mM Tris，pH 8.0，200mM NaCl和10％甘油)过HiLoad 26/60Superdex200凝胶过滤柱进一步分离纯化蛋白。
AlphaScreen实验
采用AlphaScreen试剂盒(购自Perkin Elmer)，方法如下：150μl实验体系中含1:10稀释的10×buffer(50mM MOPS，pH 7.4，50mM NaF，50mM CHAPS，0.1mg/ml bovine serum albumin)，各100nM带Biotin标签序列的HAB1和带6×His和SUMO双标签序列的PYR1/PYL1/PYL2/PYL7/GmPYL6/OsPYL2蛋白，相应浓度的(+)-ABA/0224/0304/0706/0708/0713/0715/0428/NC0F4，5μg/ml供体珠(donor beads)和受体珠(acceptor beads)(购自Perkin Elmer)，室温下避光孵育1.5小时后，置于Envision Plate Reader(购自Perkin Elmer)中按设定的AlphaScreen程序进行读数。
HAB1磷酸酶活性检测
反应体系中含50mM咪唑，pH 7.2，5mM MgCl2，0.1％β-巯基乙醇，0.5μg·ml-1BSA，100nM带Biotin标签序列的HAB1蛋白，500nM带6×His-SUMO双标签序列的PYL2受体蛋白及相应浓度的(+)-ABA/0224/0304/0706/0708/0713/0715/1028c/0428/1022B/NC0F4，室温孵育30分钟，随后加入含11个氨基酸的磷酸化多肽作为底物继续反应30分钟，该磷酸化多肽为SnRK2.6蛋白激酶的170-180位氨基酸，其中175位的磷酸化丝氨酸(序列为HSQPKpSTVGTP，购自金斯瑞)为已知的HAB1脱磷酸化靶位点。30分钟后加入显色试剂(购自BioVision)，用酶标仪(购自Molecular Device)读取650nm波长的吸收值。
基因表达分析
取全植株或叶子，用常规方法进行RNA抽提、经逆转录后进行荧光定量PCR。每种处理取3个生物学重复并进行两次实验重复，ACT7基因被用作内参。
蛋白晶体解析
结晶前，将酶切去除标签后的拟南芥PYL2和HAB1蛋白与(+)-ABA或化合物0428按1:1:5的摩尔比混合，浓缩到6mg/ml用于点晶体。采用悬滴法进行点晶体；用于结晶的孔缓冲液(well buffer)包含0.2M酒石酸钠(Di-sodium tartrate)和20％PEG 3350。一天后可看到晶体出现，约3-4天可长到100-120μm。晶体用X射线衍射并收集衍射数据，再根据相关PYR/PYL受体结构模型来解析复合物 结构。
种子萌发和土壤干旱实验
(1)种子萌发
以本发明的化合物0428和0224为例。拟南芥Col-0生态型和PYL受体三缺失突变体(pyr1；pyl1；pyl4)的种子用NaClO消毒后置于4℃春化3天，然后播种在含1μM(+)-ABA/0224化合物或0.05％DMSO(对照)的1/2MS固体培养基上，或含2μM(+)-ABA/本发明化合物0428或0.05％DMSO(对照)的1/2MS固体培养基上。每个6cm直径的培养基上同时播种两个株系，每个株系播种15-20粒种子，每种化合物设4个重复。培养基置于22℃长日照培养，在含1μM(+)-ABA/0224化合物的固体培养基上萌发的种子在播种6天后拍照，含2μM(+)-ABA/0428化合物的固体培养基上萌发的种子在播种9天后拍照。
(2)植物叶面蒸腾实验
拟南芥叶面蒸腾实验使用ABA合成突变体aba2-1。在环境胁迫条件下，该突变体中的内源ABA含量并不增加，仅为同样条件下野生型拟南芥Col-0中的1/40，因此使用该突变体可以排除内源ABA对于蒸腾实验的影响。持续浇水三周后的植物喷施含0.05％Tween-20和相应浓度的(+)-ABA/0224/0304/0706/0715/0428或0.05％Tween-20和0.05％DMSO(对照)，使用量为1.2ml/盆。大豆的叶面蒸腾实验在26℃长日照进行，在播种14天后喷施含0.1％tween-20和20μM(+)-ABA/0224/0304化合物或0.1％Tween-20和0.05％DMSO(对照)的溶液，使用量为4ml/盆；棉花的叶面蒸腾实验在播种25天后分别喷施含0.1％tween-20和20μM(+)-ABA/0224化合物或0.1％Tween-20和0.05％DMSO(对照)的溶液，使用量为4ml/盆；小麦的叶面蒸腾实验则在播种18天后分别喷施含0.1％tween-20和100μM(+)-ABA/0224化合物或0.1％Tween-20和0.05％DMSO(对照)的溶液，使用量为6ml/盆。所有植株在喷施前和喷施后每天在相同时间段使用FLIR A655sc红外热像仪成像。
(3)土壤干旱实验
拟南芥Col-0生态型种子用NaClO消毒后置于4℃春化3天后播种在1/2MS固体培养基上，生长6天后选取长势良好且大小一致的幼苗移入装满土的8×7×6cm3的花盆中。每个花盆装有相同重量的土并移入相同数目的植物(六株)以减少实验误差，所有的花盆均置于22℃短日照培养，两周后停止浇水进行干旱处理，期间每周向叶面喷洒一次含0.05％Tween-20和5μM的0224/0706/0715/0428或0.05％Tween-20和0.05％DMSO(对照)的溶液，喷施量为2ml溶液/盆，干旱过程中每天变换花盆位置以减少环境因素引起的误差，整个干旱期间总计喷施两次溶液，四周后拍照记录。
进行叶面蒸腾实验的大豆、棉花和小麦同时用于土壤干旱实验，每个花盆装有相同重量的土以减少实验误差。所有的大豆植株均在26℃长日照培养，播种14天后停止浇水，选择长势一致的植株进行干旱处理。干旱开始时向叶面喷施一次含0.1％Tween-20和20μM(+)-ABA/0224/0304或0.1％Tween-20和0.05％DMSO(对照)的溶液，喷施量为4ml/盆，同时变换花盆位置，干旱6天后复水并在复水一天后拍照。棉花干旱实验与大豆类似，播种25天后停止浇水，选择长势一致的植株进行干旱处理。干旱开始时向叶面喷施一次含0.1％Tween-20和20μM(+)-ABA/0224 或0.1％Tween-20和0.05％DMSO(对照)的溶液，此后每3天喷施一次，喷施量为4ml/盆，同时变换花盆位置，干旱6天后复水，并在复水前及复水一天后拍照。小麦的干旱实验则选取播种16天的长势一致的小麦进行干旱处理，开始干旱时向叶面喷施一次含0.1％Tween-20和100μM(+)-ABA/0224或0.1％Tween-20和0.05％DMSO(对照)的溶液，此后每3天喷施一次，喷施量为4ml/盆，同时变换花盆位置，干旱6天后拍照。
用于化合物0428的大豆和玉米的土壤干旱实验与用于化合物0224的大豆干旱实验类似，每盆只包含一株植物。所有的大豆植株均在26℃长日照培养，3组3叶后停止浇水，选择长势一致的植株进行干旱处理；玉米则在小喇叭口期停止浇水进行干旱处理。开始干旱的第一天和第二天各向叶面喷洒一次含0.05％Tween-20和50μM(+)-0428或0.05％Tween-20和0.05％DMSO(对照)的溶液，喷施量为4ml/盆，同时变换花盆位置。大豆在干旱9天后，玉米在干旱4天后复水，第二天拍照。
实施例1 化合物0224的制备
1.1制备4-甲基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000033
将3.4克1-(2-氨基苯基)乙醇和1.6克CDI加入到50毫升无水四氢呋喃中，使反应维持在室温，反应2小时；加入1M HCl溶液淬灭反应，加入乙酸乙酯萃取反应。合并有机相，用饱和氯化钠水溶液洗涤，在加入无水硫酸钠干燥有机相。减压浓缩。硅胶柱层析纯化，得淡黄色固体3.8克，收率为93％。
1HNMR(400MHz,DMSO-d6)：δ10.16(s,1H),7.25(t,1H),7.20(d,1H),7.02(t,1H),6.08(d,1H),5.49(m,1H),1.57(d,3H)ppm。
1.2制备4-甲基-1-丙基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000034
将3.0克4-甲基-1,4-二氢苯并噁唑-2-酮加入到80毫升N，N-二甲基甲酰胺中，在冰水浴下搅拌，分批加入1.05当量氢化钠，加完搅拌0.5小时。滴加1.05当量的碘丙烷，撤去冰水浴，反应12小时；加入饱和氯化铵溶液淬灭反应，加入乙酸乙酯萃取反应。合并有机相，用饱和氯化钠水溶液洗涤，在加入无水硫酸钠干燥有机相。减压蒸去溶剂和过量的碘丙烷，得到3.1克油状4-甲基-1-丙基-1,4-二氢苯并噁唑-2-酮。未进一步纯化直接进行下一步，粗收率为85％。
1.3制备4-甲基-6-硝基-1-丙基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000035
在冰水浴下将20毫升硫酸加入到盛有3.0克4-甲基-1-丙基-1,4-二氢苯并噁唑-2-酮烧瓶中，并剧烈搅拌0.5小时；用滴液漏斗缓慢滴加1.1当量的硝酸钾的硫酸溶液，维持冰水浴温度并反应1-2小时；倾倒反应液倒冰水中搅拌0.5h。过滤并用大量的水洗涤滤饼。用乙醇对粗品进行重结晶。得2.5克4-甲基-6-硝基-1-丙基-1,4-二氢苯并噁唑-2-酮，收率为74％。
1HNMR(400MHz,DMSO-d6)：δ8.27(dd,1H),8.05(d,1H),7.07(d,1H),5.47(m,1H),3.82(t,2H),1.78(m,2H),1.77(d,3H),1.04(t,3H)ppm。
1.4制备6氨基-4-甲基-1-丙基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000036
将1.7克4-甲基-6-硝基-1-丙基-1,4-二氢苯并噁唑-2-酮加入到甲醇中，加入钯碳作为催化剂。反应体系用氢气置换三次。室温搅拌8小时。反应液通过加入硅藻土的玻璃砂漏斗，滤去固体。硅胶柱层析纯化，得黄色固体1.5克，收率为90％。
1HNMR(400MHz,DMSO-d6)：δ6.77(d,1H),6.60(dd,1H),6.48(d,1H),5.27(m,1H),3.82(t,2H),3.49(s,2H),1.74(m,2H),1.63(d,3H),0.98(t,3H)ppm。
1.5制备2，3，5，6-四氟-4-甲基苄磺酰氯
Figure PCTCN2016107516-appb-000037
取2，3，5，6-四氟-4-甲基苄氯和1当量的硫脲溶于乙醇中，然后慢慢加热至回流。反应4-6小时后浓缩反应液。加入乙腈和浓盐酸。控制温度5-10℃下，在剧烈搅拌下分批加入3.5当量的亚氯酸钠。在15-20℃下反应8-16小时。加入水来终止反应。乙酸乙酯萃取三次，浓缩萃取液得淡黄色固体。未进一步纯化直接进行下一步。
1.6制备化合物0224
Figure PCTCN2016107516-appb-000038
将1.0克6-氨基-4-甲基-1-丙基-1,4-二氢苯并噁唑-2-酮和1当量的2，3，5，6-四氟-4-甲基苄磺酰氯加入到DMF中，再加入3当量碳酸钾为缚酸剂。使反应保持在室温搅拌12-16小时。反应结束加入冰水，乙酸乙酯萃取。无水硫酸钠干燥，浓缩有机相。粗品用硅胶柱层析纯化，得1.7克淡黄色固体，收率为75％。
1HNMR(400MHz,DMSO-d6)：δ10.18(s,1H),7.17-7.06(m,3H),5.36(m,1H),4.61(s,2H),3.77(m,2H),2.22(s,3H),1.61(m,2H),1.49(d,3H),0.92(t,3H)ppm。
实施例2 化合物0304的制备
2.1制备2-(2-氨基苯基)丙-2-醇
Figure PCTCN2016107516-appb-000039
将5.0克邻氨基苯乙酮加入到150毫升无水四氢呋喃中，-40摄氏度下滴加2当量的甲基碘化镁四氢呋喃溶液。使反应缓慢升到室温搅拌12小时。加入饱和氯化铵水溶液终止反应。乙酸乙酯萃取。无水硫酸钠干燥。浓缩有机相。得黄色粗品4.2克，未进一步纯化粗，收率为75％。
2.2制备4，4-二甲基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000040
将3.4克2-(2-氨基苯基)丙-2-醇和5.5克CDI加入到50毫升无水四氢呋喃中，室温反应12小时；加入1M HCl溶液淬灭反应，加入乙酸乙酯萃取反应。合并有机相，用饱和氯化钠水溶液洗涤，在加入无水硫酸钠干燥有机相。减压蒸去溶剂，得到3.7克淡黄色固体，收率为92％。
1HNMR(400MHz,CDCl3)：δ9.61(s,1H),7.25(m,1H),7.15(d,1H),7.07(m,1H),6.92(dd,1H),1.74(s,6H)ppm。
2.3制备4，4-二甲基-1-丙基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000041
将4.0克4，4-二甲基-1,4-二氢苯并噁唑-2-酮加入到80毫升N，N-二甲基甲酰胺中，在冰水浴下搅拌分批加入1.05当量氢化钠，加完搅拌0.5小时；滴加1.05当量的碘丙烷，撤去冰水浴，反应12小时；加入饱和氯化铵溶液淬灭反应，加入乙酸乙酯萃取反应,合并有机相，用饱和氯化钠水溶液洗涤，加入无水硫酸钠干燥有机相。减压蒸去溶剂和过量的碘丙烷。硅胶柱层析纯化，得到2.9克油状4，4-二甲基-1-丙基-1,4-二氢苯并噁唑-2-酮收率为60％。
1HNMR(400MHz,CDCl3)：δ7.33(m,1H),7.20-7.18(d,1H),7.10(m,1H),6.96(d,1H),3.91(t,2H),1.76(m,2H),1.71(s,6H),1.02(t,3H)ppm。
2.4制备4，4-二甲基-6硝基-1-丙基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000042
在冰水浴下将20毫升硫酸加入到盛有2.0克4，4–二甲基-1-丙基-1,4-二氢苯并噁唑-2-酮烧瓶中，剧烈搅拌0.5小时；用滴液漏斗缓慢滴加1.1当量的硝酸钾硫酸溶液，维持冰水浴温度并反应1-2小时；倾倒反应液倒冰水中搅拌0.5小时。过滤并用大量的水洗涤滤饼。用乙醇对粗品进行重结晶。得1.8克4，4-二甲基-6-硝基-1-丙基-1,4-二氢苯并噁唑-2-酮，收率为78％。
1HNMR(400MHz,CDCl3)：δ8.25(dd，1H)8.08(d,1H),7.06(d,1H),3.97(t,2H),1.80(m,2H),1.76(s,6H),1.05(t,3H)
2.5制备6-氨基-4，4-二甲基-1-丙基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000043
将1.8克4，4-二甲基-6-硝基-1-丙基-1,4-二氢苯并噁唑-2-酮加入到100毫升甲醇中，氮气氛围下加入100毫克钯碳作为催化剂。反应体系用氢气置换三次。室温搅拌8小时。反应液通过加入硅藻土的玻璃砂漏斗，滤去固体，浓缩滤液。硅胶柱层析纯化，得黄褐色固体1.4克，收率为90％。
1HNMR(400MHz,CDCl3)：δ6.76(d,1H),6.65(dd,1H),6.53(d,1H),3.84(t,2H),3.34(s,2H),1.74(m,2H),1.64(s,6H),0.99(t,3H)ppm。
2.6制备化合物0304
Figure PCTCN2016107516-appb-000044
将0.8克6-氨基-4，4-二甲基-1-丙基-1,4-二氢苯并噁唑-2-酮和1.1当量的2，3，5，6-四氟-4-甲基苄磺酰氯分批加入到DMF中，再加入3当量碳酸钾为缚酸剂。使反应保持在室温搅拌12-16小时。反应结束加入冰水，乙酸乙酯萃取，无水硫酸钠干燥。浓缩有机相，粗品用硅胶柱层析得1.1克淡黄色固体，收率为70％。
1HNMR(400MHz,CDCl3)：δ7.23(dd,1H),7.10(d,1H),6.91(d,1H),4.49(s,2H),3.89(t,2H),2.25(s,3H),1.76(m,2H),1.67(s,6H),1.03(t,3H)ppm。
实施例3 化合物0706的制备
3.1制备4-甲基-6-硝基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000045
在冰水浴下将20毫升硫酸加入到盛有3.0克4-甲基-1,4-二氢苯并噁唑-2-酮烧瓶中，并剧烈搅拌0.5小时；用滴液漏斗缓慢滴加1.1当量的硝酸钾的硫酸溶液，维持冰水浴温度并反应1-2小时；倾倒反应液倒冰水中搅拌0.5h。过滤并用大量的水洗涤滤饼。用乙醇对粗品进行重结晶。得2.5克4-甲基-6-硝基-1,4-二氢苯并噁唑-2-酮，收率为77％。
1HNMR(400MHz,DMSO-d6)：δ10.87(s,1H),8.20-8.18(d,1H),8.13(s,1H),7.07-7.05(d,1H),5.68-5.64(m,1H),1.64-1.62(d,3H)ppm。
3.2制备4-甲基-6-硝基-1-乙基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000046
将3.0克4-甲基-6-硝基-1,4-二氢苯并噁唑-2-酮加入到80毫升N，N-二甲基甲酰胺中，在冰水浴下搅拌，分批加入1.05当量氢化钠，加完搅拌0.5小时。滴加1.05当量的碘乙烷，撤去冰水浴，反应12小时；加入饱和氯化铵溶液淬灭反应，加入乙酸乙酯萃取反应。合并有机相，用饱和氯化钠水溶液洗涤，在加入无水硫酸钠干燥有机相。减压蒸去溶剂和过量的碘乙烷，得到3.1克4-甲基-6-硝基-1-乙基-1,4-二氢苯并噁唑-2-酮。未进一步纯化直接进行下一步，粗收率为85％。
3.3制备6-氨基-4-甲基-1-乙基-1,4-二氢苯并噁唑-2-酮
Figure PCTCN2016107516-appb-000047
将2.0克4-甲基-6-硝基-1-乙基-1,4-二氢苯并噁唑-2-酮加入到甲醇中，加入钯碳作为催化剂。反应体系用氢气置换三次。室温搅拌8小时。反应液通过加入硅藻土的玻璃砂漏斗，滤去固体，得1.7克固体。粗品未经进一步纯化，粗收率为89％。
3.4制备化合物0706
Figure PCTCN2016107516-appb-000048
将1.0克6-氨基-4-甲基-1-乙基-1,4-二氢苯并噁唑-2-酮和1当量的2，3，5，6-四氟-4-甲基苄磺酰氯加入到DMF中，再加入3当量碳酸钾为缚酸剂。使反应保持在室温搅拌12-16小时。反应结束加入冰水，乙酸乙酯萃取。无水硫酸钠干燥，浓缩有机相。粗品用硅胶柱层析纯化，得1.8克淡黄色固体，收率为82％。
1HNMR(400MHz,DMSO-d6)：δ10.18(s,1H),7.18-7.06(m,3H),5.42(m,1H),4.61(s,2H),3.86(q,2H),2.22(s,3H),1.48(d,3H),1.19(t,3H)ppm。
实施例4 化合物0708的制备
0708的中间体制备方法和0706相同，只是用2-碘丙烷代替1-碘乙烷。
Figure PCTCN2016107516-appb-000049
1.0克6-氨基-4-甲基-1-异丙基-1,4-二氢苯并噁唑-2-酮和1当量的2，3，5，6-四氟-4-甲基苄磺酰氯加入到DMF中，再加入3当量碳酸钾为缚酸剂。使反应保持在室温搅拌12-16小时。反应结束加入冰水，乙酸乙酯萃取。无水硫酸钠干燥，浓缩有机相。粗品用硅胶柱层析纯化，得1.6克淡黄色固体，收率为72％。
1HNMR(400MHz,DMSO-d6)：δ10.19(s,1H),7.20-7.07(m,3H),5.29(m,1H),4.61(s,2H),4.30(m,1H),2.21(s,3H),1.50-1.44(m,9H)ppm。
实施例5 化合物0713的制备
0713的中间体制备方法和0706相同，只是用1-碘异丁基代替1-碘乙烷。
Figure PCTCN2016107516-appb-000050
将1.0克6-氨基-4-甲基-1-异丁基-1,4-二氢苯并噁唑-2-酮和1当量的2，3，5，6-四氟-4-甲基苄磺酰氯加入到DMF中，再加入3当量碳酸钾为缚酸剂。使反应保持在室温搅拌12-16小时。反应结束加入冰水，乙酸乙酯萃取。无水硫酸钠干燥，浓缩有机相。粗品用硅胶柱层析纯化，得1.3克淡黄色固体，收率为63％。
1HNMR(400MHz,DMSO-d6)：δ10.14(s,1H),7.10-7.02(m,3H),5.34(q,1H),4.57(s,2H),3.69(d,2H),2.23-2.17(m,4H),1.46(d,3H),0.87-0.84(dd,6H)ppm。
实施例6 化合物0715的制备
0715的中间体制备方法和0706相同，只是用1-氟-3-碘丙烷代替1-碘乙烷。
Figure PCTCN2016107516-appb-000051
将1.0克6-氨基-4-甲基-1-(3-氟丙基)-1,4-二氢苯并噁唑-2-酮和1当量的2，3，5，6-四氟-4-甲基苄磺酰氯加入到DMF中，再加入3当量碳酸钾为缚酸剂。使反应保持在室温搅拌12-16小时。反应结束加入冰水，乙酸乙酯萃取。无水硫酸钠干燥，浓缩有机相。粗品用硅胶柱层析纯化，得1.4克淡黄色固体，收率为71％。
1HNMR(400MHz,DMSO-d6)：δ10.19(s,1H),7.18-7.06(m,3H),5.42(q,1H),4.61(s,2H),4.61(t,1H),4.50(t,1H)，3.95(t,2H),2.22(s,3H),2.06-1.94(m,2H),1.51(d,3H)ppm
实施例7 化合物0428的制备
7.1制备1,4-二氢-2H-3,1-苯并噁嗪-2-酮
Figure PCTCN2016107516-appb-000052
将3.0克2-氨基苯甲醇和1.6克尿素加入到80毫升DMF中，升温到150℃，反应12小时；加入饱和氯化钠溶液淬灭反应，加入乙酸乙酯萃取反应三次。合并有机相，用饱和氯化钠水溶液和2N盐酸洗涤除去未反应的2-氨基苯甲醇和尿素，在加入无水硫酸钠干燥有机相。减压蒸干得到3.0克1,4-二氢-2H-3,1-苯并噁嗪-2-酮。HPLC检测纯度为92％，无需进一步纯化直接进行下一步，收率为83％。1HNMR(400MHz，DMSO-d6)：δ5.27(s，2H),6.85-7.27(m,4H),10.15(s,1H)ppm
7.2制备1-丙基-1,4-二氢-2H-3,1-苯并噁嗪-2-酮
Figure PCTCN2016107516-appb-000053
将2.0克1,4-二氢-2H-3,1-苯并噁嗪-2-酮加入到80毫升N，N-二甲基甲酰胺中，在冰水浴下搅拌分批加入1.05当量氢化钠，加完搅拌0.5小时；滴加1.05当量的碘丙烷，撤去冰水浴，反应12小时；加入饱和氯化铵溶液淬灭反应，加入乙酸乙酯萃取反应。合并有机相，用饱和氯化钠水溶液洗涤，在加入无水硫酸钠干燥有机相。减压蒸去溶剂和过量的碘丙烷，得到2.2克油状1-丙基-1,4-二氢-2H-3,1-苯并噁嗪-2-酮。HPLC检测纯度为92％，无需进一步纯化直接进行下一步，收率为88％。
7.3制备6-硝基-1-丙基-1,4-二氢-2H-3,1-苯并噁嗪-2-酮
Figure PCTCN2016107516-appb-000054
在冰水浴下将20毫升硫酸加入到盛有2克1-丙基-1,4-二氢-2H-3,1-苯并噁嗪-2-酮烧瓶中，并剧烈搅拌0.5h；用滴液漏斗缓慢滴加1.1当量的硝酸钾硫酸溶液，维持冰水浴温度并反应1-2小时；倾倒反应液到冰水中搅拌0.5h。过滤并用大量的水洗涤滤饼。红外灯下干燥。用乙醇对粗品进行重结晶。得1.8克6-硝基-1-丙基1,4-二氢-2H-3,1-苯并噁嗪-2-酮，收率为77％。1HNMR(400MHz，DMSO-d6)：δ0.93(t，3H),1.63(m,2H),3.85(t,2H),5.39(s,2H),7.37(m,1H),8.22(m,2H)ppm。
7.4制备6-氨基-1-丙基-1,4-二氢-2H-3,1-苯并噁嗪-2-酮
Figure PCTCN2016107516-appb-000055
将1.8克6-硝基-1-丙基1,4-二氢-2H-3,1-苯并噁嗪-2-酮加入到甲醇中，再加入钯碳作为催化剂。反应体系用氢气置换三次。室温搅拌8小时。反应液通过加入硅藻土的玻璃砂漏斗，滤去固体。浓缩滤液得1.4克6-氨基-1-丙基-1,4-二氢-2H-3,1-苯并噁嗪-2-酮.未进一步纯化直接进行下一步，粗收率为90％。
7.5制备2,3,5,6-四氟-4-甲基苄磺酰氯
Figure PCTCN2016107516-appb-000056
取1.0克2,3,5,6-四氟-4-甲基氯苄和1当量的硫脲溶于40毫升乙醇中，然后慢慢加热至回流。反应4-6h后浓缩反应液，得白色固体。加入10毫升乙腈和4毫升浓盐酸。控制温度5-10℃下，在剧烈搅拌下分批加入2.25克亚氯酸钠。在15-20 ℃下反应8-16小时。加入水来终止反应。乙酸乙酯萃取三次。浓缩萃取液得2,3,5,6-四氟-4-甲基苄磺酰氯。未进一步纯化直接进行下一步。
7.6制备化合物0428
Figure PCTCN2016107516-appb-000057
将1.0克6-氨基-1-丙基-1,4-二氢-2H-3,1-苯并噁嗪-2-酮和1.2当量的2,3,5,6-四氟-4-甲基苄磺酰氯加入到20毫升DMF中，再加入3当量碳酸钾为缚酸剂。使反应保持在室温搅拌12-16小时。反应结束加入冰水，乙酸乙酯萃取。无水硫酸钠干燥。浓缩有机相。粗品用硅胶柱层析得1.5克化合物0428，收率为70％。
1HNMR(400MHz，DMSO-d6)：δ0.92(t，3H),1.60(m,2H),2.27(s,3H),3.77(t,2H),4.60(s,2H),5.17(s,2H),7.10-7.18(m,3H),10.19(s,1H)ppm。
实施例8 化合物1022B的制备
8.1制备1-丙基-2(1H)-喹啉酮
Figure PCTCN2016107516-appb-000058
将4.0克2-羟基喹啉加入到100毫升DMF中，在冰水浴下搅拌,分批加入1.1当量氢化钠，加完保温搅拌0.5小时；滴加1.1当量的碘丙烷，撤去冰水浴，反应12小时；加入饱和氯化铵溶液淬灭反应，乙酸乙酯萃取反应。合并有机相，用饱和氯化钠水溶液洗涤，加入无水硫酸钠干燥有机相。减压蒸去溶剂和过量的碘丙烷，得到油状粗品。通过硅胶层析柱分离，得到无色油状液体1-丙基-2(1H)-喹啉酮3.3克，收率为62％。1HNMR(400MHz，DMSO-d6)：δ0.95(t，3H),1.62(m,2H),4.18(t,2H),6.63(d,1H),7.25(t,1H),7.60(m,2H),7.72(d,1H),7.90(d,1H)ppm。
8.2制备6-硝基-1-丙基-2(1H)-喹啉酮
Figure PCTCN2016107516-appb-000059
在冰水浴下将40毫升硫酸加入到装有2.0克1-丙基-2(1H)-喹啉酮的烧瓶中，并剧烈搅拌0.5小时；用滴液漏斗缓慢滴加15毫升1.1当量的硝酸钾硫酸溶液，维持冰水浴温度并反应1-2小时；倾倒反应液到冰水中搅拌0.5小时。过滤并用大量的水洗涤滤饼。红外灯下干燥。用乙醇对粗品进行重结晶。得1.7克6-硝基-1-丙基-2(1H)-喹啉酮，收率为72％。1HNMR(400MHz，DMSO-d6)：δ0.95(t，3H),1.63(m,2H),4.24(t,2H),6.76(d,1H),7.76(d,1H),8.12(d,1H),8.35(d,1H),8.71(s,1H)ppm。
8.3制备6-氨基-1-丙基-2(1H)-喹啉酮
Figure PCTCN2016107516-appb-000060
将1.7克6-硝基-1-丙基-2(1H)-喹啉酮加入到80毫升甲醇中，再加入钯碳作为催化剂。反应体系用氢气置换三次。室温搅拌2小时。反应液通过加入硅藻土的玻璃砂漏斗，滤去固体。浓缩滤液得1.4克6-氨基-1-丙基-2(1H)-喹啉酮。粗品无需进一步纯化，收率为90％。1HNMR(400MHz，DMSO-d6)：δ0.92(t，3H),1.60(m,2H),4.08(t,2H),5.08(s,2H),6.46(d,1H),6.79(s,1H),6.94(d,1H),7.29(d,1H),7.66(d,1H)ppm。
8.4制备对甲基卤代苄磺酰氯
Figure PCTCN2016107516-appb-000061
取1.0克对甲基苄溴与1当量的硫脲加入到40毫升乙醇中，然后慢慢加热至回流，溶液转为澄清。反应4-6小时后浓缩反应液,得白色固体。加入10毫升乙腈和4毫升浓盐酸。控制温度在5-10℃，剧烈搅拌下分批加入2.25克的亚氯酸钠。在15-20℃下反应8-16小时。加水来终止反应。乙酸乙酯萃取三次。浓缩萃取液得对甲基苄磺酰氯。粗品未经纯化直接用于下一步。
8.5制备化合物1022B
Figure PCTCN2016107516-appb-000062
将1.0克6-氨基-1-丙基-2(1H)-喹啉酮和1.2当量的对甲基苄磺酰氯加入到20毫升DMF中，再加入3当量碳酸钾为缚酸剂。使反应在室温搅拌12-16小时。结束后加入冰水，乙酸乙酯萃取三次，合并有机相并用无水硫酸钠干燥。浓缩有机相，粗品用硅胶柱层析得1.3克化合物1022B，收率为70％。
1HNMR(400MHz，DMSO-d6)：δ0.95(t，3H),1.62(m,2H),2.27(s,3H),4.18(t,2H),4.42(s,2H),6.60(d,1H),7.12-7.17(m,4H),7.40(d,1H),7.45(s,1H),7.55(d,1H),7.83(d,1H)ppm；13CNMR(100MHz,DMSO-d6)δ11.53,20.99,21.54,43.35,57.34,116.08,119.20,121.17,122.14,124.01,126.89,129.46,131.34,133.02,135.97,138.12,139.45,161.12ppm。
实施例9 化合物NC0F4的制备
9.1制备2-氨甲基-4-硝基苯胺
Figure PCTCN2016107516-appb-000063
将5.0克2-氰基-4-硝基苯胺加入到200毫升干燥四氢呋喃中，在冰水浴下搅拌滴加3.5当量硼烷四氢呋喃溶液，加毕，室温搅拌过夜；缓慢滴加入饱和氯化铵溶液淬灭反应，加入二氯甲烷萃取混合液。合并有机相，用饱和氯化钠水溶液洗涤，在加入无水硫酸钠干燥有机相。减压蒸去溶剂和过量的碘丙烷，得到4.2克2-氨甲基-4-硝基苯胺，收率83％。1HNMR(400MHz，DMSO-d6)：δ8.05(d，1H),7.88(dd,1H),6.70-6.59(m,3H),3.63(s,2H)ppm。
9.2制备6-硝基-3,4-二氢喹唑啉-2(1H)-酮
Figure PCTCN2016107516-appb-000064
甲基-4-硝基苯胺加入到250毫升干燥四氢呋喃中，搅拌下加入1.5当量CDI。加毕，升温至回流并搅拌过夜；减压蒸去四氢呋喃，加入水和二氯甲烷萃取混合液。合并有机相，用饱和氯化钠水溶液洗涤，在加入无水硫酸钠干燥有机相。得到6-硝基-3,4-二氢喹唑啉-2(1H)-酮,收率74％。1HNMR(400MHz，DMSO-d6)：δ9.78(s，1H),8.07(s,1H),8.04(m,1H),7.17(d,1H),6.93(d,1H),4.43(s,2H)ppm。
9.3制备1-丙基-6-硝基-3,4-二氢喹唑啉-2(1H)-酮
Figure PCTCN2016107516-appb-000065
合成方法同实施例7.2，只是用6-硝基-3,4-二氢喹唑啉-2(1H)-酮代替1,4-二氢-2H-3,1-苯并噁嗪-2-酮。粗品未经进一步纯化，直接进行下一步，粗收率为56％。
9.4制备1-丙基-6-氨基-3,4-二氢喹唑啉-2(1H)-酮。
Figure PCTCN2016107516-appb-000066
合成方法同实施例7.4，只是用1-丙基-6-硝基-3,4-二氢喹唑啉-2(1H)-酮替代6-硝基-1-丙基1,4-二氢-2H-3,1-苯并噁嗪-2-酮。粗品未经进一步纯化，粗收率为87％
9.5制备NC0F4
Figure PCTCN2016107516-appb-000067
合成方法同实施例7.6，只是用1-丙基-6-氨基-3,4-二氢喹唑啉-2(1H)-酮代替6-氨基-1-丙基-1,4-二氢-2H-3,1-苯并噁嗪-2-酮。1HNMR(400MHz，DMSO-d6)：δ10.0(s，1H),7.06-6.90(m,4H,),4.56(s,2H),4.20(s,2H),3.69(s,2H),2.24(s,3H),1.52(m,2H),0.89(t,3H)ppm。
实施例10 化合物1028c的制备
10.1制备N-(1-(2-氨基苯基)乙叉)-叔丁基亚磺酸胺
Figure PCTCN2016107516-appb-000068
将2.0克2-氨基苯乙酮和1当量的叔丁基亚磺酰胺，加到2当量的钛酸四乙酯中，充入氮气，封管加热到75℃并搅拌16小时；冷却至室温，倒入冰盐水中，淬灭反应，加入乙酸乙酯萃取反应。合并有机相，用饱和氯化钠水溶液洗涤，加入无水硫酸钠干燥有机相。减压浓缩，硅胶柱层析纯化，得淡黄色固体2.7克，收率为74％。1HNMR(400MHz,DMSO-d6)：δ7.66(d,1H),7.43(s,2H),7.20(t,1H),6.75(d,1H),6.58(t,1H),2.68(s,3H),1.18(s,9H)ppm。
10.2制备N-(1-(2-氨基苯基)乙基)-叔丁基亚磺酸胺
Figure PCTCN2016107516-appb-000069
将2.5克制备N-(1-(2-氨基苯基)乙叉)-叔丁基亚磺酸胺溶解在绝对无水四氢呋喃中，溶液控制在-78℃，在氮气氛围下缓慢滴加硼烷二甲基硫醚络合物四氢呋喃溶液，加毕搅拌3小时；滴加饱和食盐水淬灭反应，加入乙酸乙酯萃取反应。合并有机相，用饱和氯化钠水溶液洗涤，在加入无水硫酸钠干燥有机相。减压浓缩，硅胶柱层析纯化，的淡黄色固体2.3克，收率为91％。1HNMR(400MHz,DMSO-d6)：δ7.16(d,1H),6.96(t,1H),6.64(d,1H),6.56(t,1H),5.37(d,1H),4.95(s,2H),4.40(m,1H),4.14(d,3H)1.11(s,9H)ppm。
10.3制备2-(1-氨基乙基)苯胺
Figure PCTCN2016107516-appb-000070
将2.2克N-(1-(2-氨基苯基)乙基)-叔丁基亚磺酸胺溶解在甲醇和4M盐酸(1：1)中，溶液搅拌过夜；减压浓缩，调pH值直至碱性。加入饱和食盐水/乙酸乙酯萃取反应。合并有机相，用饱和氯化钠水溶液洗涤，在加入无水硫酸钠干燥有机相。减压浓缩，得1.2克，收率为91％。1HNMR(400MHz,DMSO-d6)：δ7.05(d,1H),6.89(t,1H),6.56(d,1H),6.50(t,1H),5.27(s,2H),4.03(q,1H),3.37(in water peak)1.27(d,3H)ppm。
10.4制备4-甲基-3,4-二氢喹唑啉-2(1H)-酮
Figure PCTCN2016107516-appb-000071
将1.2克2-(1-氨基乙基)苯胺溶解于70毫升无水四氢呋喃中，分批加入1.2当量三光气(BTC)，室温反应12小时；加入1M HCl溶液淬灭反应，加入乙酸乙酯萃取反应。合并有机相，用饱和氯化钠水溶液洗涤，加入无水硫酸钠干燥有机相。减压蒸去溶剂，得到1.2克淡黄色固体，收率为82％。1HNMR(400MHz,DMSO-d6)： δ9.06(s,1H),7.12-6.84(m,4H),6.77(d,1H),4.45(m,1H),1.31(d,3H)ppm。
10.5制备4-甲基-1-丙基-1,4-二氢喹唑啉-2(1H)-酮
Figure PCTCN2016107516-appb-000072
将1.2克4-甲基-3,4-二氢喹唑啉-2(1H)-酮加入到80毫升N，N-二甲基甲酰胺中，在冰水浴下搅拌分批加入1.05当量氢化钠，加毕搅拌0.5小时；滴加1.05当量的碘丙烷，撤去冰水浴，反应12小时；加入饱和氯化铵溶液淬灭反应，加入乙酸乙酯萃取反应,合并有机相，用饱和氯化钠水溶液洗涤，加入无水硫酸钠干燥有机相。减压蒸去溶剂和过量的碘丙烷。得到1.4克油状物，未经进一步纯化，收率为92％。
10.6制备4-甲基-6-硝基-1-丙基-1,4-二氢喹唑啉-2(1H)-酮
Figure PCTCN2016107516-appb-000073
在冰浴下将20毫升硫酸加入到盛有1.2克4-甲基-1-丙基-3,4-二氢喹唑啉-2(1H)-酮烧瓶中，剧烈搅拌0.5小时；用滴液漏斗缓慢滴加1.1当量的硝酸钾硫酸溶液，维持冰水浴温度并反应1-2小时；倾倒反应液倒冰水中搅拌0.5小时。过滤并用大量的水洗涤滤饼。用乙醇对粗品进行重结晶。得1.1克4-甲基-6-硝基-1-丙基-1,4-二氢喹唑啉-2(1H)-酮，收率为72％。1HNMR(400MHz,DMSO-d6)：δ8.11(dd，1H)8.10(s,1H),7.54(d,J＝1.6Hz,1H),7.18(d,J＝8.8Hz,1H),4.62(m,1H),3.81(m,2H),1.55(m,2H),1.33(d,3H),0.90(t,3H)ppm。
10.7制备6-氨基-4-甲基-1-丙基-1,4-二氢喹唑啉-2(1H)-酮
Figure PCTCN2016107516-appb-000074
将1.1克4-甲基-6-硝基-1-丙基-1,4-二氢喹唑啉-2(1H)-酮加入到100毫升甲醇中，氮气氛围下加入40毫克钯碳作为催化剂。反应体系用氢气置换三次。室温搅拌8小时。反应液通过加入硅藻土的玻璃砂漏斗，滤去固体，浓缩滤液。硅胶柱层析纯化，得黄褐色固体1.2克，收率为93％。
1HNMR(400MHz,DMSO-d6)：δ6.75(s,1H),6.65(d,J＝8.8Hz,1H),6.46(dd,1H),6.37(d,J＝2.0Hz,1H),4.75(s,2H),4.26(m,1H),3.67(m,2H),1.51(m,2H),1.25(d,J＝6.4Hz,3H)0.86(t,J＝7.2Hz,3H)ppm。
10.8制备1028c
Figure PCTCN2016107516-appb-000075
将1.0克6-氨基-4-甲基-1-丙基-1,4-二氢喹唑啉-2(1H)-酮和1.1当量的2，3，5，6-四氟-4-甲基苄磺酰氯分批加入到DMF中，再加入3当量三乙胺(TEA)为缚酸剂。使反应保持在室温搅拌12-16小时。反应结束加入冰水，乙酸乙酯萃取，无水硫酸钠干燥。浓缩有机相，粗品用硅胶柱层析得1.1克淡黄色固体，收率为70％。
1HNMR(400MHz,DMSO-d6)：δ10.0(s,1H),7.07-7.04(m,2H),6.98(d,J＝2.4Hz,1H),6.92(d,J＝8.8Hz,1H),4.56(s,2H),4.35(m,1H),3.75(m,2H),3.03(s,3H),1.52(m,2H),1.25(d,J＝6.4Hz,3H),0.87(t,J＝7.2Hz,3H)ppm。
实施例11 化合物0224，0304，0706,0708,0713,0715,1028c，0428，1022B和NC0F4的体外活性测试实验
体外生化实验表明，多个本发明化合物作为高效的PYL受体激动剂，与多个PYL受体具有高亲和性的结合能力，同时促进PYL受体结合并抑制PP2C蛋白磷酸酶活性。
11.1体外生化实验和PP2C蛋白磷酸酶活性测试
结果如图1a-1g所示。以SnRK2.6磷酸化多肽为底物的HAB1蛋白磷酸酶活性实验表明，化合物0224，0304，0706，0708，0713，0715，1028c，0428，1022B和NC0F4均可促进PYL2受体可与PP2C蛋白磷酸酶(HAB1)结合，从而抑制HAB1对于SnRK2.6磷酸化多肽的去磷酸化作用，且在低浓度下大多数化合物的效果优于或显著优于同浓度的ABA。
11.2 AlphaScreen实验
用AlphaScreen技术检测多个本发明化合物对于PYL受体和PP2C蛋白磷酸酶(HAB1)结合的促进能力。
实验结果表明，化合物0224和0304与PYL受体-HAB1蛋白磷酸酶复合物的结合能力均存在剂量依赖效应。对于拟南芥PYR1,PYL1,PYL2和PYL7受体，化合物0224和0304均具有显著优于ABA的受体亲和性，其中0224和0304与PYR1受体-HAB1蛋白磷酸酶复合物的EC50值分别为ABA的1/40和1/24(图2a)，与PYL1受体-HAB1蛋白磷酸酶复合物的EC50值分别为ABA的1/8和1/7(图2b),与PYL2受体-HAB1蛋白磷酸酶复合物的EC50值分别为ABA的1/29和1/15(图2c),与PYL7受体-HAB1蛋白磷酸酶复合物的EC50值则均为ABA的1/10(图2d)。化合物0706，0708，0713，0715，0428，1022B和NC0F4同样表现出优于ABA的PYL2受体亲和性，其中0706，0708，0713和0715的EC50值约为ABA的1/5-1/8(图2e)。化合物0428的EC50值约为ABA的1/8(图2f)，1022B的EC50值则比ABA低1个数量级(图2g)，且PYL2受体与HAB1的结合能力存在上述化合物的剂量依赖效应。化合物NC0F4则表现出优于ABA的PYR1受体亲和性，其EC50值约为ABA的1/11(图2h)。
上述结果表明，化合物0224，0304，0706，0708，0713，0715，1028c，0428，1022B和NC0F4是比ABA更为高效的PYL受体激动剂。
此外，使用大豆GmPYL6(拟南芥PYL2的同源基因)和水稻OsPYL2(拟南芥PYL2的同源基因)与拟南芥AtHAB1的实验表明，化合物0428与大豆GmPYL6蛋白和水稻OsPYL2蛋白同样具有显著高于ABA的亲和性，其EC50值分别只有ABA的约1/14和1/3(图13)。
上述结果表明，上述多个本发明化合物是一系列比ABA等现有化合物更为高效的PYL受体激动剂。
并且，当体外测试的本发明其他化合物的浓度区间在0.01-100μM时，本发明的化合物均表现出显著的PYR/PYL受体亲和性。
实施例12 多个本发明化合物(如0428，0224，0304，0706，0715等)的萌发抑制和抗旱活性测试
12.1对拟南芥种子萌发的抑制效果
结果如图3和图15所示。以化合物0224和0428为例。1μM化合物0224或2μM化合物0428均能够抑制Col-0生态型种子的萌发，而无法抑制PYR/PYL三突变体pyr1；pyl1；pyl4种子的萌发。上述结果表明，本发明的化合物(如化合物0224和0428)的萌发抑制效应是因为其激活了植物内在的ABA信号通路，而非对植物种子产生了毒性。
12.2对拟南芥、大豆、棉花和小麦叶面蒸腾作用的抑制
本实验中，利用红外摄像机观察记录叶面的温度变化，从而反映出植物的蒸腾作用强弱。蒸腾作用越强的植物，叶面温度越低。
拟南芥叶面蒸腾实验结果如图4所示。5μM ABA,0224化合物(图4a)或0304化合物(图4b)喷施拟南芥aba2-1后，叶面温度均高于DMSO对照组，这意味着化合物处理的植物蒸腾作用减弱，且喷施0224或0304化合物四天后的植株叶片温度仍显著高于DMSO对照组，甚至在浓度降至2μM或1μM后，喷施相应浓度的0224化合物四天后的植物叶片温度仍显著高于DMSO对照组(图4b)，而喷施5μM ABA四天后的植株叶片温度已降至喷施前的水平。上述结果表明化合物0224和0304对拟南芥叶面蒸腾作用的抑制效果优于ABA，且对蒸腾作用的抑制存在剂量效应，其中0224的效果更佳。使用5μM化合物0706或0715喷施拟南芥aba2-1后，叶面温度同样较对照组(DMSO)处理显著增加，其持续时间与0224相当(图4c)。而经过5μM/2μM/1μM 0428化合物处理一天的叶面温度也均高于DMSO对照组，意味着化合物处理的植物蒸腾作用减弱，同时表明0428化合物对于蒸腾作用的抑制也存在剂量效应(图4d)。
大豆、棉花和小麦的叶面蒸腾实验结果如图5和图6所示，对大豆进行的叶面蒸腾抑制实验表明，喷施20μM的化合物0224或0304的植株，两天后其叶面温度仍显著高于喷施DMSO的对照组，表明此时大豆叶面的蒸腾效应仍受到抑制，而喷施相同浓度ABA的植株叶面温度已与对照组无异(图5)。对棉花进行的叶面蒸腾抑制实验则表明，喷施20μM的化合物0224的植株，两天后其叶面温度仍显著高于喷施DMSO的对照组，而喷施相同浓度ABA的植株叶面温度已与对照组无异(图6)。对小麦进行的叶面蒸腾抑制实验表明，喷施100μM 的化合物0224或ABA的植株，一天后其叶面温度仍显著高于喷施DMSO的对照组，表明小麦叶面的蒸腾效应受到抑制(图16)。这一结果表明化合物0224不仅在双子叶作物大豆和棉花中可以抑制叶面蒸腾作用，在单子叶作物小麦中同样有抑制叶面蒸腾作用的效果。上述结果表明化合物0224和0304在大豆中以及化合物0224在棉花和小麦中也存在与拟南芥同样的抑制叶面蒸腾作用的效果。
12.3对拟南芥、大豆、棉花、玉米和小麦抗旱性的增强
在土壤中生长两周的拟南芥Col-0生态型停止给水，干旱期间每周向叶面喷洒一次含5μM的化合物0224/0706/0715或0.05％DMSO(对照)的溶液，喷施量为2ml溶液/盆，共喷施两次，溶液中同时添加了0.05％(v/v)的表面活性剂Tween-20以增强喷剂对于叶片表皮的穿透作用。经过四周的干旱处理后，喷施DMSO的对照组均已旱死，而喷施5μM化合物0224/0706/0715的植物仍能存活(图7)。同样方法进行干旱处理以及(+)-ABA或0428处理的拟南芥在干旱两周后拍照记录。如图12所示，由于浓度过低，喷施5μM(+)-ABA的植物长势与喷施DMSO的对照组无异，但喷施5μM 0428的拟南芥长势明显优于喷施DMSO的对照组和喷施5μM(+)-ABA的植物。
分别播种14天的大豆和25天的棉花，选取相同大小的植株进行土壤干旱实验。大豆在开始干旱后一次性喷施含20μM的ABA/0224/0304化合物或0.05％DMSO(对照)的水溶液，棉花则在开始干旱后每3天喷施一次含20μM的ABA/0224化合物或0.05％DMSO(对照)的水溶液，上述溶液中均添加了0.1％(v/v)的表面活性剂Tween-20以增强喷剂对于叶片表皮的穿透作用。干旱6天后复水，喷施20μM化合物0224或0304化合物的大豆(图8)以及喷施20μM化合物0224的棉花(图9)在复水后的长势均明显优于喷施DMSO的对照组以及喷施相同浓度ABA的植株。
在另一组实验中，选取相同大小的3组3叶期大豆或小喇叭口期玉米植株进行土壤干旱实验，开始干旱后连续两天每天喷施一次含50μM的0428化合物或0.05％DMSO(对照)的水溶液，溶液中同样添加了0.1％(v/v)的表面活性剂Tween-20。玉米和大豆分别经过四天和九天的干旱处理后复水，喷施50μM的化合物0428的大豆(图14a)和玉米(图14b)在复水后的长势均明显优于喷施DMSO的对照组。
在另一组实验中，播种16天的小麦，选取相同大小的植株进行土壤干旱实验。小麦在开始干旱后每3天喷施一次含100μM的ABA/0224化合物或0.1％DMSO(对照)的水溶液，上述溶液中均添加了0.1％(v/v)的表面活性剂Tween-20以增强喷剂对于叶片表皮的穿透作用。干旱6天后对照组小麦已萎蔫，而经100μM化合物0224或ABA处理的小麦均仍保持直立，长势显著优于对照组(DMSO)(图16)。
上述结果表明本发明化合物在双子叶和单子叶植物中均有显著增强抗旱性的效果。
实施例13 化合物1022B可诱导ABA响应的胁迫相关基因的表达
本发明人分析了外源添加化合物1022B对于植物基因表达的影响。
基因表达分析的结果表明，化合物1022B可诱导ABA响应的胁迫相关基因的表达，且表达水平大多可以达到或高于相同浓度的外源ABA所诱导的表达水平(图10)。10μM的化合物1022B处理后，10天大的野生型拟南芥(Col-0)苗期植株中，4个已知的受ABA诱导的与环境胁迫相关基因(COR15a,COR47,RAB18和RD29b)的表达量显著增加，显著超过10μM ABA处理相同时间后的水平。
结果表明，化合物1022B对于多数环境胁迫相关基因的诱导效果要显著优于ABA。
实施例14 PYL2-0428-HAB1复合物的结构
采用通用方法中所述的蛋白晶体解析方法，检测了本发明化合物0428所形成的PYL2-0428-HAB1复合物晶体结构。复合物晶体分辨率为2.4埃，对照为ABA，两种复合物晶体的二维结构局部示意图如图11a和图11b所示。
从图11a和图11b看出，0428存在于PYL2的口袋结构中。ABA结构上的四个氧原子可通过多个水分子与PYL2口袋结构以及HAB1的多个氨基酸残基形成氢键。化合物0428磺酰氨基团上的氧原子和氮原子以及喹啉环上的氧原子同样可以形成氢键，此外，对二甲苯上的卤素取代基(氟原子)还可与PYL2口袋结构内的氨基酸残基形成氢键，进一步增强化合物0428与PYL2受体的亲和性。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (10)

  1. 一种式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体，
    Figure PCTCN2016107516-appb-100001
    式中，
    R1为H、卤素、C1-C3烷基、或C1-C3卤代烷基；
    R2为H、卤素、C1-C3烷基、或C1-C3卤代烷基；
    R3为H、卤素、C1-C3烷基、或C1-C3卤代烷基；
    R4为H、卤素、C1-C3烷基、或C1-C3卤代烷基；
    R5为卤素、C1-C3烷基、C1-C3卤代烷基、SF5或C3-C8环烷基；
    R6为取代或未取代的C1-C7烷基、取代或未取代的C2-C7链烯基、取代或未取代的C2-C7链炔基、取代或未取代的C3-C7环烷基、或取代或未取代的-Ra-O-Rb，其中，Ra为C1-C2亚烷基而Rb为H、C1-C3烷基；并且所述的取代为具有一个或多个选自下组的取代基：卤素、-ORb、-CN、-N(Rb)2、和硝基；
    R7选自下组：H、取代或未取代的C1-C6烷基、取代或未取代的C2-C6链烯基、取代或未取代的C2-C6链炔基、取代或未取代的C3-C8环烷基、取代或未取代的C5-C10杂环基、Rc-C(O)-、-ORb、-CN、和-N(Rb)2；Rc选自下组：羟基、巯基、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基；其中，所述杂环基含有1至2个选自N、O、S的杂原子，并且所述的取代为具有一个或多个选自下组的取代基：卤素、-ORb、-CN、-N(Rb)2、和硝基；
    R8、R9、R10各自独立地选自下组：
    (i)H；
    (ii)取代或未取代的C1-C6烷基、C3-C8烷氧基、取代或未取代的C3-C8环烷基、卤素、Rc-C(O)-、-OH、-NH2；R11选自下组：羟基、巯基、取代或未取代的C1-C6烷基、取代或未取代的C1-C6烷氧基；其中所述的取代为具有一个或多个选自下组的取代基：卤素、-ORb、-CN、-N(Rb)2、和硝基；
    R11为H、C1-C3烷基、或C1-C3卤代烷基；
    X为CR12、NR13、O、或S，其中R12选自下组：H、卤素、C1-C3烷基、C2-C3链烯基、C2-C3链炔基、C1-C3卤代烷基、或其组合；R13为无，或选自下组：H、卤素、C1-C3烷基、C2-C3链烯基、C2-C3链炔基、C1-C3卤代烷基、或其组合；
    m＝1或2；
    Figure PCTCN2016107516-appb-100002
    表示单键或双键；
    附件条件是，X为CR12时，
    Figure PCTCN2016107516-appb-100003
    为双键。
  2. 如权利要求1所述的式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体，其特征在于，所述化合物具有式Ia结构：
    Figure PCTCN2016107516-appb-100004
    式Ia中，R1-R10、m的定义如上所述。
  3. 如权利要求1所述的式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体，其特征在于，所述化合物具有式Ib结构：
    Figure PCTCN2016107516-appb-100005
    式Ib中，R1-R10、R13、m、
    Figure PCTCN2016107516-appb-100006
    的定义如上所述。
  4. 如权利要求1所述的式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体，其特征在于，所述化合物具有式Ic结构：
    Figure PCTCN2016107516-appb-100007
    式Ic中，R1-R10、m的定义如上所述。
  5. 如权利要求1所述的式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体，其特征在于，所述化合物具有式Id结构：
    Figure PCTCN2016107516-appb-100008
    式Id中，R1-R10、R12、m的定义如上所述。
  6. 如权利要求1所述的式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体，其特征在于，所述化合物选自下组：
    Figure PCTCN2016107516-appb-100009
    Figure PCTCN2016107516-appb-100010
    Figure PCTCN2016107516-appb-100011
    Figure PCTCN2016107516-appb-100012
    在另一优选例中，所述化合物选自下组：
    Figure PCTCN2016107516-appb-100013
  7. 一种权利要求1所述式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体的用途，其特征在于，用于制备农用制剂或组合物，所述农用制剂或组合物用于(i)增强植物抗逆性；(ii)制备ABA受体的激动 剂；和/或(iii)制备种子萌发抑制剂。
  8. 一种农用制剂，其特征在于，所述农用制剂包括：
    (i)权利要求1所述的式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体；和
    (ii)农业上可接受的载体。
  9. 一种增强植物抗逆性的方法，其特征在于，给所述植物施用本发明权利要求1所述的式I化合物、或其盐、或其光学异构体、或其外消旋体、或其溶剂化物、或其前体或施用权利要求8所述的农用制剂。
  10. 一种式I化合物或其盐的制法，其特征在于，包括步骤：
    (a)在惰性溶剂中，将化合物I-A与化合物I-S2进行反应，从而形成式I化合物；
    Figure PCTCN2016107516-appb-100014
    上述各式中，R1、R2、R3、R4、R5、R6、R7、R8、R9、R10、R11、m、X、
    Figure PCTCN2016107516-appb-100015
    如权利要求1所定义。
PCT/CN2016/107516 2015-12-28 2016-11-28 高抗逆性的植物生长调节剂及其制法和用途 Ceased WO2017114052A1 (zh)

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