Nitrification inhibitors

Abstract

The present invention generally relates to nitrification inhibitors and compositions comprising nitrification inhibitors. The present invention also relates to the use of the nitrification inhibitors and compositions for application to fertilizers, plants, agricultural areas (e.g., soil or pasture) to reduce or inhibit the oxidation of ammonium nitrogen to nitrite and nitrate nitrogen, such as oxidation of fertilizers based on ammonia or urea.

Description

Technical Field

[0001] The present invention generally relates to nitrification inhibitors and compositions comprising nitrification inhibitors. The present invention also relates to the use of the nitrification inhibitors and compositions for application to fertilizers, plants, agricultural areas (e.g., soil or pasture) to reduce or inhibit the oxidation of ammonium nitrogen to nitrite (salt) and nitrate (salt) nitrogen, such as the oxidation of fertilizers based on ammonia or urea. Background Art

[0002] High application of nitrogen fertilizer is common in agricultural systems to achieve optimal yields. However, due to the well-known low nitrogen use efficiency (NUE), this practice leads to the release of reactive nitrogen species into the surrounding environment. Plants rarely absorb more than 50% of the applied fertilizer nitrogen. In Australia, NUE falls anywhere between 6 and 59%, depending on the crop type; globally, NUE has remained at about 50% since the 1980s (Chen, D. et al., Australian Journal of Soil Research, 2008, 46, 289–301; Rowlings, AW et al., Agriculture, Ecosystems and Environment, 2016, 216, 216–225). The remaining nitrogen is easily lost from the plant / soil system via volatilization of ammonia (NH3), nitrate (NO3 - ) leaching, and gaseous emissions from denitrification.

[0003] A related issue is the loss that results in the release of nitrous oxide (N2O), a global warming agent 300 times more potent than carbon dioxide (CO2), which also catalyzes the destruction of stratospheric ozone. Over the past three decades, atmospheric N2O concentrations have increased at a rate of 0.73 ppb per year, with the use of nitrogen fertilizers being a major contributor (Ciais, P. et al., Carbon and Other Biogeochemical Cycles. In: Climate Change 2013: The Physical Science Basis. Contribution of Working Group I to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change, Cambridge University Press, Cambridge, United Kingdom and New York, NY, USA).

[0004] Ammonium in soil (NH4+ ) (directly applied, or indirectly produced by microbial conversion of nitrogen fertilizers) is rapidly oxidized to nitrite (NO2 - ) and subsequently oxidized to NO3 - . Then, NO3 - Undergoes denitrification, where NO3 - Sequentially reduced to NO2 - , nitric oxide (NO), N2O, and finally N2. With high NO3 - Soils with nitrogen content are at risk of losing nitrogen through: NO3 - Self-leaching or gaseous loss of NO and N2O from incomplete denitrification. - Concentration scenarios are desirable to mitigate these losses.

[0005] Use fertilizers modified with nitrification inhibitors to mitigate NH4 + To NO3 - The conversion of nitrification is a strategy to increase NUE. Nitrification inhibitors inhibit nitrifying microorganisms in the soil and increase NH4 + residence time and reduction of leaching (NO3 - ) and denitrification (N2O, NO x , N2). The Intergovernmental Panel on Climate Change (IPCC) has also recommended the use of nitrification inhibitors to mitigate N2O emissions. Of the many compounds identified as nitrification inhibitors, the most widely studied commercial products are based on one of three chemicals: dicyandiamide (DCD, AlzChem AG), 2-chloro-6-(trichloromethyl)-pyridine (Nitrapyrin or N-Serve, Dow Chemical Co.), and 3,4-dimethylpyrazole phosphate (DMPP or ENTEC, BASF AG; the active compound is 3,4-dimethylpyrazole (DMP)). The effectiveness (potency) of these inhibitors varies widely and appears to be affected by environmental conditions and soil properties such as pH, water content / rainfall, temperature, and soil type.

[0006] DMPP is often considered one of the more promising nitrification inhibitor candidates because it has undergone extensive toxicological testing, is effective at low concentrations, and has low mobility in soil due to its positive charge (Zerulla, W. et al., Biology and Fertility of Soils, 2001, 34, 79-84). Although DMPP is by far the most promising inhibitor, it has been found to have widely varying inhibitory activity for reducing leaching and N2O emissions in field studies - from no effect to up to 70% inhibition, for reasons that are not well understood. DMPP has been shown to have little to no effect on improving crop / biomass yields, and is therefore not an attractive option from an economic perspective for farmers, who would ideally compensate for the cost of higher fertilizers with increased yields.

[0007] It is also known that DMPP inhibitory activity is negatively correlated with temperature, with a significant reduction in activity observed in a relatively small temperature window. Studies have shown that at a temperature of 35°C, DMPP remains effective for only one week (Mahmood, T. et al., Soil Research, 2017, 55, 715-722).

[0008] It has also been reported that low pH soil conditions severely reduce DMPP activity, possibly due to a switch from autotrophic bacteria (targeted by DMPP) to heterotrophic bacteria (dominant under these acidic conditions) (Barth, G. et al., Biology and Fertility of Soils, 2001, 34, 98-102; Xi, R. et al., AMB Express, 2017, 7, 129). Attempts to circumvent some of these problems include using succinic acid to reformulate the active 3,4-dimethylpyrazole (DMP) core to produce an isomeric mixture of 2-(N-3,4-dimethylpyrazole)succinic acid and 2-(N-4,5-dimethylpyrazole)succinic acid, called DMPSA. It is believed that DMPSA is metabolized to the active DMP core once applied to the soil, resulting in a longer life in the soil.

[0009] Therefore, it is necessary to develop new nitrification inhibitors to solve the above-mentioned deficiencies. Summary of the invention

[0010] The present invention is based on the discovery that substituted 1,2,3-triazoles are effective nitrification inhibitors with low volatility.

[0011] Thus, in one aspect, the present invention provides a method for reducing nitrification in soil, comprising treating the soil with a compound of formula (I) or an agriculturally acceptable salt thereof:

[0012]

[0013] in

[0014] R 1 and R 2 Independently selected from: optionally substituted -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 ;

[0015] R 3 is H, or is selected from: optionally substituted -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 ;

[0016] R 4 is selected from: -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl; and

[0017] R 5 and R 6 Independently selected from: H, -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl.

[0018] In another aspect, the present invention provides a composition for reducing nitrification comprising a compound of formula (I) as defined above and at least one agriculturally acceptable adjuvant or diluent.

[0019] In a further aspect, the present invention provides a fertilizer comprising a urea or ammonium based fertilizer and a compound of formula (I) as defined herein.

[0020] In yet another aspect, the present invention provides a compound of formula (II) or an agriculturally acceptable salt thereof:

[0021]

[0022] in

[0023] R 1 and R 2 Independently selected from: optionally substituted -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 ;

[0024] R 3is H, or is selected from: optionally substituted -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 ;

[0025] R 4 is selected from: -C2-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl; and

[0026] R 5 and R 6 Independently selected from: H, -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl;

[0027] Provided that the compound is not:

[0028] 1-Butyl-4-pentyl-1H-1,2,3-triazole;

[0029] 1,4-Butyl-1H-1,2,3-triazole;

[0030] 4-Butyl-1H-1,2,3-triazole-1-acetic acid ethyl ester;

[0031] 1-Butyl-4-(α,α-dimethylmethanol)-1H-1,2,3-triazole;

[0032] 4-Butyl-1H-1,2,3-triazole-1-propylamine;

[0033] 4,5-bis(hydroxymethyl)-1H-1,2,3-triazole-1-acetic acid ethyl ester; or

[0034] 1,4-Dipropyl-1H-1,2,3-triazole.

[0035] In a further aspect of the present invention, the present invention provides a compound of formula (IIa) or an agriculturally acceptable salt thereof:

[0036]

[0037] in

[0038] R 1 -C1-C substituted by 10 Alkyl: one or more hydroxyl groups, -C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl group containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl group is optionally substituted by: one or more C1-C 10 Alkyl, oxo (oxy), hydroxy, C1-C4 alkoxy-, or amino; or

[0039] R 1 Selected from: -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C2-C 10 Alkyl C(O)OC1-C4 alkyl, -C1-C 10 Alkyl C(O)OC2-C4 alkenyl, -C1-C 10 Alkyl C(O)OC2-C4 alkynyl, -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino;

[0040] R 2 Selected from: -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino;

[0041] R 3 is H, or is selected from: -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino;

[0042] R 4 is selected from: -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl; and

[0043] R 5 and R 6 independently selected from: H, -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl; or

[0044] R 1 is -CH2C(O)OC1-C4 alkyl, and R 2 and R 3 Each is -CH2OC(O)C1-C4 alkyl.

[0045] These and other aspects of the present invention will become more apparent to the skilled artisan upon reading the following detailed description in conjunction with the appended examples and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] The invention will herein be described, by way of example only, with reference to the following non-limiting drawings, in which:

[0047] Figure 1NH4 measured in Horsham soils incubated (cultivated) at 25°C (A, B) and 35°C (C, D) after treatment with + -N(A,C) and NO x - -N(B, D) concentration: (NH4)2SO4[●], (NH4)2SO4+H-DMPP[■], (NH4)2SO4+13[◆], (NH4)2SO4+14[○], (NH4)2SO4+16[□]. Nitrification inhibition by NH4 + -Slow decrease of N and NO x — The slow increase of N indicates that

[0048] Figure 2 Figure 2 shows the calculated NO after 28 days incubation in Horsham soil at 25°C and 35°C. x - -N production rate (mg NO x - -N / kg soil / day). All samples were treated with 100 mg N kg -1 The ratio of nitrification to (NH4)2SO4 was treated with the fertilizer (NH4)2SO4. The values ​​presented are means (n=3); the errors are the standard error of the mean. x - -N production rate indicated.

[0049] Figure 3 Measured NH4 in Dahlen soils incubated at 25°C (A, B) and 35°C (C, D) after treatment with + -N(A,C) and NO x - -N(B, D) concentration: (NH4)2SO4[●], (NH4)2SO4+H-DMPP[■], (NH4)2SO4+13[◆], (NH4)2SO4+16[□]. Nitrification inhibition by NH4 + -Slow decrease of N and NO x - The slow increase of -N indicates.

[0050] Figure 4 Figure 2 shows the calculated NO after 28 days of incubation in Dahlen soil (pH 7.3) at 25°C and 35°C. x - -N production rate (mg NO x --N / kg soil / day). All samples were treated with 100 mg N kg -1 The ratio of nitrification was treated with the fertilizer (NH4)2SO4. The values ​​presented are means (n=3); the errors are the standard error of the mean. x - -N production rate indicated.

[0051] Figure 5 Measured NH4 in Dahlen soils incubated at 25°C (A, B) and 35°C (C, D) after treatment with + -N(A,C) and NO x - -N(B, D) concentration: (NH4)2SO4[●], (NH4)2SO4+H-DMPP[■], (NH4)2SO4+18[○], (NH4)2SO4+23[◇]. Nitrification is inhibited by the slow reduction of NH4+-N and NO x --The slow increase of N indicates.

[0052] Figure 6 Measured NH4 in South Johnstone soils incubated at 25°C (A, B) and 35°C (C, D) after treatment with + -N(A,C) and NO x - -N (B, D) concentration: (NH4)2SO4[●], (NH4)2SO4+H-DMPP[■], (NH4)2SO4+3[◆], (NH4)2SO4+16[□], (NH4)2SO4+18[○]. Nitrification is inhibited by the slow reduction of NH4+-N and NO x — The slow increase of N indicates that

[0053] Figure 7 Illustrate the results of soil TLC leaching of the inhibitor compound DMP and compound 16 in Dahlen soil (A) or South Johnstone soil (B). Higher R f Values ​​indicate higher degrees of leachability through the soil profile. DETAILED DESCRIPTION

[0054] Mono-, di-, and tri-substituted 1,2,3-triazoles are studied as potential nitrification inhibitors. Substituted 1,2,3-triazoles are considered good candidates because they are readily available synthetically using copper-catalyzed click chemistry pathways and have been used as pharmacophores in medicine and pharmaceuticals due to their wide range of biological activities. Variation of the substitution pattern at the 1,4 and / or 5 positions allows optimization of any inhibitory activity. It is believed that changing the substituents and substitution pattern can enable the nitrification inhibitors to be tailored to specific soils (such as acidic, neutral and alkaline soils) and different climatic conditions.

[0055] In one embodiment, the present invention provides a method for reducing nitrification in soil, comprising treating the soil with a compound of formula (I) or an agriculturally acceptable salt thereof:

[0056]

[0057] in

[0058] R 1 and R 2 Independently selected from: optionally substituted -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 ;

[0059] R 3 is H, or is selected from: optionally substituted -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 ;

[0060] R 4 is selected from: -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl; and

[0061] R 5 and R 6 Independently selected from: H, -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl.

[0062] In one embodiment, with respect to Formula (I), R 1 and R 2 Independently selected from: -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6), which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino;

[0063] R 3 is H, or is selected from: -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino;

[0064] R 4 is selected from: -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl; and

[0065] R 5 and R 6 Independently selected from: H, -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl.

[0066] In this specification, unless otherwise defined, the term "optionally substituted by" is understood to mean that the group may or may not be further substituted by one or more groups selected from the group consisting of hydroxy, alkyl, alkoxy, alkoxycarbonyl, alkoxycarbonyloxy, alkenyl, alkenyloxy, alkynyl, alkynyloxy, amino, aminoacyl, amido, thio, arylalkyl, arylalkoxy, aryl, aryloxy, amido, carboxy, cyano, halogen, nitro, sulfo, phosphonyl, phosphoramido, phosphinyl, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclyloxy, trihalomethyl, pentafluoroethyl, trifluoromethoxy, difluoromethoxy, trifluoro Methylthio, trifluorovinyl, mono- and di-alkylamino, mono- and di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-heteroarylamino, mono- and di-heterocyclylamino, asymmetric di-substituted amines having different substituents selected from alkyl, aryl, heteroaryl, and heterocyclyl, mono- and di-alkylamide, mono- and di-(substituted alkyl)amide, mono- and di-arylamide, mono- and di-heteroarylamide, mono- and di-heterocyclylamide, asymmetric di-substituted amides having different substituents selected from alkyl, aryl, heteroaryl, and heterocyclyl.

[0067] As used herein, the term "alkyl" (used alone or in compound words) means a straight chain or branched alkyl group. Prefixes such as "C2-C 10 " is used to indicate the number of carbon atoms in the alkyl group (in this case 2 to 10). Examples of straight chain and branched alkyl groups include: methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, hexyl, heptyl, 5-methylheptyl, 5-methylhexyl, octyl, nonyl, decyl, undecyl, dodecyl, and docosyl (C 22 ).

[0068] As used herein, the term "alkenyl" (alone or in compound words) means a straight-chain or branched hydrocarbon residue containing at least one carbon-carbon double bond, including olefinic mono-, di-, or polyunsaturated alkyl groups as defined above. 2- C 20 " is used to indicate the number of carbon atoms in the alkenyl group (in this case 2 to 20). Examples of alkenyl groups include vinyl, allyl, 1-methylvinyl, butenyl, isobutenyl, 3-methyl-2-butenyl, 1-pentenyl, 1-hexenyl, 3-hexenyl, 1-heptenyl, 3-heptenyl, 1-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 1-decenyl, 3-decenyl, 1,3-butadienyl, 1,4-pentadienyl, 1,3-hexadienyl, 1,4-hexadienyl, and 5-docenyl (C 22 ).

[0069] As used herein, the term "alkynyl" (used alone or in compound words) refers to a straight-chain or branched hydrocarbon residue containing at least one carbon-carbon triple bond. Prefixes such as "C2-C 20 " is used to indicate the number of carbon atoms in the alkenyl group (in this case 2 to 20).

[0070] As used herein, the term "amino" refers to a nitrogen atom substituted with, for example, hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, or a combination thereof.

[0071] As used herein, the term "amido" refers to an amide group, i.e., a group of formula -C(O)NH2. The group is bonded to the remainder of the molecule via the carbonyl carbon atom. The nitrogen atom may also be substituted, for example, by an alkyl, alkenyl, alkynyl, aryl, heteroaryl, or a combination thereof.

[0072] The term "aryl" refers to an aromatic monocyclic group (e.g., phenyl) or a polycyclic group (e.g., tricyclic, bicyclic, such as naphthalene, anthracenyl, phenanthrenyl). The aryl group may also be fused or bridged with an alicyclic or heterocyclic ring to form a polycyclic ring (e.g., tetralin, methylenedioxyphenyl).

[0073] As used herein, the term "heteroaryl" represents a monocyclic or bicyclic ring, typically having up to 7 atoms in each ring, wherein at least one ring is aromatic and contains 1 to 4 heteroatoms selected from O, N, and S. Heteroaryl groups within the scope of this definition include, but are not limited to: benzimidazole (also known as benzoimadazole), acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl, benzofuranyl, quinolyl, isoquinolinyl, oxazolyl, isoxazolyl, indoiyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrahydroquinoline. As with the definition of heterocycle below, "heteroaryl" is also understood to include the N-oxide derivative of any nitrogen-containing heteroaryl. In cases where the heteroaryl substituent is bicyclic and one ring is non-aromatic or contains no heteroatoms, it is understood that attachment is via the aromatic ring or the heteroatom-containing ring, respectively.

[0074] The term "heteroatom" includes atoms of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, sulfur, and phosphorus.

[0075] The term "alkoxy" includes substituted and unsubstituted alkyl, alkenyl, and alkynyl groups covalently attached to an oxygen atom. Examples of alkoxy groups include methoxy, ethoxy, isopropyloxy (isopropoxy), propoxy, butoxy, and pentyloxy groups, and may include cyclic groups such as cyclopentyloxy.

[0076] In one embodiment, the method as defined above comprises co-treating the soil with a fertilizer.

[0077] In another embodiment, the method as defined above is effective for reducing nitrification in soil at elevated ambient temperatures, for example at ambient temperatures between about 25°C and about 50°C, such as between about 30°C and about 45°C.

[0078] It will be appreciated that fertilizers can be formulated to contain a mixture of minerals and nutrients, wherein the source of nitrogen simply provides one of the many minerals and nutrients present in the fertilizer. The fertilizer can be a nitrogen-based fertilizer. The nitrogen-based fertilizer can be a fertilizer based on ammonium, ammonium nitrate or urea, or include ammonia, ammonium, nitrate or urea (or can contain all three forms, as in the case of urea ammonium nitrate). The nitrogen-based fertilizer can be an organic fertilizer or an inorganic fertilizer. The organic fertilizer can include animal waste (animal feces, animal waste). In one embodiment, the fertilizer includes an ammonium-based fertilizer, or is composed of an ammonium-based fertilizer. In another embodiment, the fertilizer includes a urea-based fertilizer, or is composed of a urea-based fertilizer.

[0079] In one embodiment, the fertilizer is an inorganic fertilizer. These may be fertilizers containing ammonium or urea. Examples of this type of ammonium-containing fertilizers are: NPK fertilizers, calcium ammonium nitrate, ammonium sulphate nitrate, ammonium sulphate, or ammonium phosphate. In a specific embodiment, the ammonium-containing fertilizer is selected from anhydrous ammonia, ammonium sulphate, urea, ammonium nitrate, ammonium phosphate, and mixtures thereof.

[0080] The fertilizer may be coated or impregnated with the nitrification inhibitor or its formulation. The fertilizer may be in the form of granules, crystals or powders incorporating the nitrification inhibitor or its formulation. The fertilizer may be a liquid fertilizer including the nitrification inhibitor or its formulation. It will be appreciated that other forms of fertilizer may be used.

[0081] Thus, in one embodiment, the present invention provides a fertilizer as defined above, wherein the urea or ammonium based fertilizer is in the form of granules and said compound of formula (I) is coated on said granules.

[0082] In a further embodiment, the method as defined above comprises co-treating the soil with a urease inhibitor.

[0083] Currently, there is only one commercially available urease inhibitor, N-(n-butyl)thiophosphoric acid triamide (NBPT, marketed as Agrotain). Unfortunately, this inhibitor has a limited lifetime in soil. The major degradation pathway in acidic and slightly alkaline soils is chemical hydrolysis, while in more alkaline soils microbial degradation becomes dominant.

[0084] Most soil conditions will benefit from a fertilizer containing both urease and a nitrification inhibitor. In addition, recent studies have shown that while nitrification inhibitors are effective in reducing N2O emissions from a variety of agricultural systems, they can increase NH3 emissions under some conditions. These issues highlight the importance of using both urease and nitrification inhibitors in mitigating nitrogen losses. Currently, commercial products containing both urease and nitrification inhibitors are limited because production is hampered by the challenges of combining acid-sensitive NBPT with acidic DMPP.

[0085] In one embodiment, there is provided a fertilizer as defined above, wherein the urea or ammonium based fertilizer is in the form of granules and the compound of formula (I) and the urease inhibitor are coated on the granules.

[0086] In one embodiment, the present invention provides a compound of formula (II) or an agriculturally acceptable salt thereof:

[0087]

[0088] in

[0089] R 1and R 2 Independently selected from: optionally substituted -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 ;

[0090] R 3 is H, or is selected from: optionally substituted -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 ;

[0091] R 4 is selected from: -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl; and

[0092] R 5 and R 6 Independently selected from: H, -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl;

[0093] Provided that the compound is not:

[0094] 1-Butyl-4-pentyl-1H-1,2,3-triazole;

[0095] 1,4-Butyl-1H-1,2,3-triazole;

[0096] 4-Butyl-1H-1,2,3-triazole-1-acetic acid ethyl ester;

[0097] 1-Butyl-4-(α,α-dimethylmethanol)-1H-1,2,3-triazole;

[0098] 4-Butyl-1H-1,2,3-triazole-1-propylamine;

[0099] 4,5-bis(hydroxymethyl)-1H-1,2,3-triazole-1-acetic acid ethyl ester; or

[0100] 1,4-Dipropyl-1H-1,2,3-triazole.

[0101] In some preferred embodiments of the present invention, and with respect to general formula (II), one or more of the following preferred embodiments apply:

[0102] (a)R 1 is C1-C substituted by 10 Alkyl: one or more hydroxyl groups, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl group containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl group is optionally substituted by: one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0103] (b)R 1 Selected from: -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C2-C 10 Alkyl C(O)OC1-C4 alkyl, -C1-C 10 Alkyl C(O)OC2-C4 alkenyl, -C1-C 10 Alkyl C(O)OC2-C4 alkynyl, -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0104] (c)R 1 is C1-C substituted by 10 Alkyl: a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by: one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0105] (d) R 1 is a C1-C 10 alkyl.

[0106] (e)R 1 is a C1-C 10 alkyl.

[0107] (f)R 1 is a -C1-C4 alkoxy-substituted 10 alkyl.

[0108] (g)R 1 C2-C 10 alkenyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0109] (h)R 1 C2-C 10 Alkynyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0110] (i)R 1 C2-C 10 Alkyl C (O) OC1-C4 alkyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0111] (j)R 1 C1-C 10 Alkyl C (O) OC2-C4 alkenyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0112] (k)R 1 C1-C 10 Alkyl C (O) OC2-C4 alkynyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0113] (l)R 1 C2-C 10 Alkenyl C(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0114] (m)R 1 C2-C 10 Alkynyl C(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0115] (n)R 1 C1-C 10 AlkylOC(O)R 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0116] (o)R 1 C2-C 10 Alkenyl OC(O)R 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0117] (p)R 1 C2-C 10 AlkynylOC(O)R 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0118] (q)R 1 C1-C 10 Alkyl OC(O)OR 4, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0119] (r)R 1 C2-C 10 Alkenyl OC(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0120] (s)R 1 C2-C 10 Alkynyl OC(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0121] (t)R 1 C1-C 10 Alkyl C(O)N(R 5 R 6 ), which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0122] (u)R 1 C2-C 10 Alkenyl C(O)N(R 5 R 6 ), which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0123] (v)R 1 C2-C 10 Alkynyl C(O)N(R 5 R 6 ), which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0124] (w)R 1 C1-C 10 Alkyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0125] (x)R 1 C2-C 10 Alkenyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0126] (y)R 1 C2-C 10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0127] (z)R 1 C3-C 10Alkyl (O) OC1-C4 alkyl.

[0128] (aa)R 2 Selected from -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 and-C2-C 10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0129] (ab)R 2 C1-C10 alkyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0130] (ac)R 2 For unsubstituted C1-C 10 alkyl.

[0131] (ad)R 2 For unsubstituted -C1-C 10 AlkylOC(O)R 4 .

[0132] (ae)R 2 is a C1-C 10 alkyl.

[0133] (af)R 2 C2-C 10 alkenyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0134] (ag)R 2 C2-C 10 Alkynyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0135] (ah)R 2 C1-C 10 Alkyl C(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0136] (ai)R 2 C2-C 10 Alkenyl C(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0137] (aj)R 2 C2-C 10 Alkynyl C(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0138] (ak)R 2 C1-C 10 AlkylOC(O)R 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0139] (al)R 2 C2-C 10 Alkenyl OC(O)R 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0140] (am)R 2 C2-C 10 AlkynylOC(O)R 4, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0141] (an)R 2 C1-C 10 Alkyl OC(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0142] (ao)R 2 C2-C 10 Alkenyl OC(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0143] (ap)R 2 C2-C 10 Alkynyl OC(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0144] (aq)R 2 C1-C 10 Alkyl C(O)N(R 5 R 6 ), which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0145] (ar)R 2 C2-C 10 Alkenyl C(O)N(R 5 R 6 ), which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0146] (as)R 2 C2-C 10 Alkynyl C(O)N(R 5 R 6 ), which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0147] (at)R 2 C1-C 10 Alkyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0148] (au)R 2 C2-C 10 Alkenyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0149] (av)R 2 C2-C10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0150] (aw)R 3 is H, or is selected from: -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0151] (ax)R 3 C2-C 10 alkyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0152] (ay)R 3 -C1-C substituted by hydroxyl 10 alkyl.

[0153] (az)R 3 C2-C 10 alkenyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0154] (ba)R 3 C2-C 10 Alkynyl, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0155] (bb)R 3 C1-C 10 Alkyl C(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0156] (bc)R 3 C2-C 10 Alkenyl C(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0157] (bd)R 3 C2-C 10 Alkynyl C(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0158] (be)R 3 C1-C 10 AlkylOC(O)R 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0159] (bf)R 3 C2-C 10 Alkenyl OC(O)R 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0160] (bg)R 3 C2-C 10 AlkynylOC(O)R 4, which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0161] (bh)R 3 C1-C 10 Alkyl OC(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0162] (bi)R 3 C2-C 10 Alkenyl OC(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0163] (bj)R 3 C2-C 10 Alkynyl OC(O)OR 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0164] (bk)R 3 C1-C 10 Alkyl C(O)N(R 5 R 6 ), which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0165] (bl)R 3 C2-C 10 Alkenyl C(O)N(R 5 R 6 ), which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0166] (bm)R 3 C1-C 10 Alkyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0167] (bn)R 3 C2-C 10 Alkenyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0168] (bo)R 3 C2-C 10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4 alkoxy-, or amino.

[0169] (bp)R 3 For unsubstituted -C1-C10 AlkylOC(O)R 4 .

[0170] (bq)R 4 Selected from C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl.

[0171] (br)R 4 It is a C1-C4 alkyl group.

[0172] (bs)R 4 It is ethyl.

[0173] (bt)R 5 and R 6 Independently selected from: H, C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl.

[0174] (bu)R 5 and R 6 One of them is H, and the other is C1-C4 alkyl, C2-C4 alkenyl, or C2-C4 alkynyl.

[0175] (bv)R 1 is -CH2C(O)OC1-C4 alkyl, and R 2 and R 3 Each is -CH2OC(O)C1-C4 alkyl.

[0176] Therefore, in one aspect, the present invention provides a compound of formula (II) represented by formula (IIa) or an agriculturally acceptable salt thereof:

[0177]

[0178] in

[0179] R 1 C1-C 10 Alkyl, which is substituted by one or more hydroxyl groups, -C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino; or

[0180] R 1 Selected from: -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C2-C 10 Alkyl C(O)OC1-C4 alkyl, -C1-C 10 Alkyl C(O)OC2-C4 alkenyl, -C1-C 10Alkyl C(O)OC2-C4 alkynyl, -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino;

[0181] R 2 Selected from: -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino;

[0182] R 3 is H, or is selected from: -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 Alkyl C(O)OR 4 , -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 AlkylOC(O)R4 , -C2-C 10 Alkenyl OC(O)R 4 , -C2-C 10 AlkynylOC(O)R 4 , -C1-C 10 Alkyl OC(O)OR 4 , -C2-C 10 Alkenyl OC(O)OR 4 , -C2-C 10 Alkynyl OC(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), -C1-C 10 Alkyl NR 5 C(O)R 6 , -C2-C 10 Alkenyl NR 5 C(O)R 6 , and -C2-C 10 Alkynyl NR 5 C(O)R 6 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino;

[0183] R 4 is selected from: -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl; and

[0184] R 5 and R 6 is independently selected from: H, -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl; or

[0185] R 1 is -CH2C(O)OC1-C4 alkyl, and R 2 and R 3 Each is -CH2OC(O)C1-C4 alkyl.

[0186] In a further embodiment, with respect to Formula (IIa), R1 Selected from: C2-C 10 Alkenyl, C2-C 10 Alkynyl, -C2-C 10 Alkyl C(O)OC1-C4 alkyl, -C1-C 10 Alkyl C(O)OC2-C4 alkenyl, -C1-C 10 Alkyl C(O)OC2-C4 alkynyl, -C2-C 10 Alkenyl C(O)OR 4 , -C2-C 10 Alkynyl C(O)OR 4 , -C1-C 10 Alkyl C(O)N(R 5 R 6 ), -C2-C 10 Alkenyl C(O)N(R 5 R 6 ), and -C2-C 10 Alkynyl C(O)N(R 5 R 6 ), which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino;

[0187] R 2 Selected from: -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10 Alkynyl, -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , and -C2-C 10 AlkynylOC(O)R 4 , which is optionally substituted by one or more amino, hydroxy, C1-C4 alkoxy-, or a 3-10 membered monocyclic or fused bicyclic heteroaryl containing one or more heteroatoms selected from N, O, and S, wherein the heteroaryl is optionally substituted by one or more C1-C 10 Alkyl, oxo, hydroxy, C1-C4alkoxy-, or amino;

[0188] R 3 is H, or is selected from: -C1-C 10 Alkyl, -C2-C 10 Alkenyl, -C2-C 10Alkynyl, -C1-C 10 AlkylOC(O)R 4 , -C2-C 10 Alkenyl OC(O)R 4 , and -C2-C 10 AlkynylOC(O)R 4 , which is optionally substituted with: one or more amino, hydroxy, or C1-C4 alkoxy groups;

[0189] R 4 is selected from C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl; and

[0190] R 5 and R 6 Independently selected from: H, C1-C4 alkyl, C2-C4 alkenyl, and C2-C4 alkynyl.

[0191] In another embodiment, the compound of formula (IIa) or an agriculturally acceptable salt thereof is selected from:

[0192] 4-Butyl-1H-1,2,3-triazole-1-butyric acid ethyl ester (5);

[0193] 2-[3-[4,5-Bis(hydroxymethyl)-1H-1,2,3-triazolyl]propyl]-isoindoline-1,3-dione (7);

[0194] 2-[3-[4,5-(acetic acid methyl ester)-1H-1,2,3-triazole]propyl]-isoindoline-1,3-dione (8);

[0195] 4,5-Bis(hydroxymethyl)-1H-1,2,3-triazole-1-butyric acid ethyl ester (9);

[0196] 4,5-Bis(methyl acetate)-1H-1,2,3-triazole-1-butyric acid ethyl ester (10);

[0197] 4,5-Bis(methyl acetate)-1H-1,2,3-triazole-1-acetic acid ethyl ester (11);

[0198] 1-Butyl-4-propyl-1H-1,2,3-triazole (13);

[0199] 1-(2-methoxyethyl)-4-butyl-1H-1,2,3-triazole (14);

[0200] 4-propyl-1H-1,2,3-triazole-1-ethanol (15);

[0201] 1-(3-Butyn-1-yl)-4-propyl-1H-1,2,3-triazole (17);

[0202] 1-(2-Propene-1-yl)-4-propyl-1H-1,2,3-triazole (18);

[0203] 2-(4-propyl-1H-1,2,3-triazol-1-yl)-acetic acid ethyl ester (19);

[0204] Prop-2-en-1-yl 2-(4-propyl-1H-1,2,3-triazol-1-yl)-acetate (20);

[0205] Prop-2-en-1-yl 2-(4-propyl-1H-1,2,3-triazol-1-yl)-acetamide (21);

[0206] Prop-2-yn-1-yl 2-(4-propyl-1H-1,2,3-triazol-1-yl)-acetate (22); and

[0207] Prop-2-yn-1-yl 2-(4-propyl-1H-1,2,3-triazol-1-yl)-acetamide (23).

[0208] It will be appreciated that the compounds of the present invention may exist in multiple equivalent tautomeric forms. For clarity, the compounds have been described as single tautomers, although all such tautomeric forms are contemplated to be within the scope of the present invention.

[0209] Some structures of the compounds of the present invention may include asymmetric carbon atoms. Therefore, it will be understood that the isomers (e.g., all enantiomers, stereoisomers, rotational isomers, tautomers, diastereomers, or racemates) produced by such asymmetry are included within the scope of the present invention. The present invention includes all these stereoisomeric forms within its scope, separated (e.g., in enantiomeric separation), or combined (including racemic mixtures and diastereomeric mixtures).

[0210] The skilled person will appreciate that there are a range of techniques available for making racemic, enantiomerically enriched, or enantiomerically pure forms of the achiral compounds of the invention. For example, enantiomerically enriched or enantiomerically pure forms of the compounds may be made by stereoselective synthesis and / or by using chromatography or selective recrystallization techniques.

[0211] The compounds of the present invention may be in crystalline (crystal) form, may be oil, or may be solvates (e.g., hydrates), and it is intended that all forms are within the scope of the present invention. The term "solvate" is a complex of variable stoichiometry formed by a solute (in this invention, a compound of the present invention) and a solvent. Preferably, such a solvent should not interfere with the biological activity of the solute. By way of example, the solvent may be water, acetone, ethanol, or acetic acid. Methods of solvation are generally known in the art.

[0212] The compounds of the present invention having at least one basic center can form acid addition salts. Acid addition salts can be prepared from inorganic and organic acids. Examples of inorganic acids include: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Examples of organic acids include: acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

[0213] The compounds of the present invention having at least one acidic group can form base addition salts. Base addition salts can be prepared from inorganic and organic bases. Corresponding counterions derived from inorganic bases include sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Organic bases include primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, and cyclic amines, including isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, triethanolamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucosamine, theobromine, purines, piperazine, piperidine, and N-ethylpiperidine.

[0214] In a further aspect, there is provided a composition for reducing nitrification in soil, the composition comprising a compound of formula (I) as defined herein and at least one agriculturally acceptable adjuvant or diluent.

[0215] The compounds according to the invention can be used as nitrification inhibitors in unmodified form, but are usually formulated into compositions in various ways using formulation aids such as carriers, solvents and surface-active substances. The formulations can be in various physical forms, for example in the form of dusting powders, gels, wettable powders, water-dispersible granules, water-dispersible tablets, effervescent pellets, emulsifiable concentrates, microemulsifiable concentrates, oil-in-water emulsions, oil-flowables, aqueous dispersions, oil dispersions, suspoemulsions, capsule suspensions, emulsifiable granules, soluble liquids, water-soluble concentrates (with water or water-miscible organic solvents as carriers), impregnated polymer films, or in other known forms. Such formulations can be used directly or diluted before use. Dilution can be carried out, for example, using a diluent selected from, but not limited to, water, liquid fertilizers, micronutrients, biological organisms, oils, or solvents.

[0216] The formulation can be prepared by mixing the nitrification inhibitor of the present invention with a formulation adjuvant to obtain a composition in the form of a finely divided solid, granules, solution, dispersion or emulsion. The nitrification inhibitor can also be formulated with other adjuvants, such as finely divided solids, mineral oils, oils of plant or animal origin, modified oils of plant or animal origin, organic solvents, water, surface-active substances, or combinations thereof.

[0217] The nitrification inhibitor may also be contained in very fine microcapsules. The microcapsules contain the active ingredient in a porous carrier so that the nitrification inhibitor can be released into the environment in a controlled amount (e.g., slowly released (sustained release)). Microcapsules typically have a diameter of 0.1 to 500 microns. They contain the active ingredient in an amount of about 25 to 95% by weight of the capsule weight. The active component may be in the form of a monolithic solid, in the form of fine particles in a solid or liquid dispersion, or in the form of a suitable solution. The encapsulating membrane may include, for example, natural or synthetic rubber, cellulose, styrene / butadiene copolymers, polyacrylonitrile, polyacrylates, polyesters, polyamides, polyureas, polyurethanes or chemically modified polymers and starch xanthates, or other polymers known to those skilled in the art. Alternatively, very fine microcapsules may be formed in which the active ingredient is contained in a solid matrix of a base material in the form of finely dispersed particles, while the microcapsules themselves are not encapsulated.

[0218] Formulation aids suitable for preparing the compositions according to the invention are known in the art. As liquid carriers, use may be made of: water, toluene, xylene, petroleum ether, vegetable oils, acetone, methyl ethyl ketone, cyclopentane (tetramethylene sulfone), cyclohexanone, anhydrides, acetonitrile, acetophenone, amyl acetate, 2-butanone, butylene carbonate, chlorobenzene, cyclohexane, cyclohexanol, alkyl esters of acetic acid, diacetone alcohol, 1,2-dichloropropane, diethanolamine, p-diethylbenzene, diethylene glycol, diethylene glycol rosin acid ester, diethylene glycol butyl ether, diethylene glycol ethyl ether ... Ethylene glycol methyl ether, N,N-dimethylformamide, dimethyl sulfoxide, 1,4-dioxane, dipropylene glycol, dipropylene glycol methyl ether, dipropylene glycol dibenzoate, dipropylene glycol (diproxitol), alkyl pyrrolidone, ethyl acetate, 2-ethylhexanol, ethylene carbonate, 1,1,1-trichloroethane, 2-heptanone, α-pinene, d-limonene, ethyl lactate, ethylene glycol, ethylene glycol butyl ether, ethylene glycol methyl ether, γ-butyrolactone, glycerol, Glyceryl acetate, glyceryl diacetate, glyceryl triacetate, hexadecane, hexylene glycol, isoamyl acetate, isobornyl acetate, isooctane, isophorone, cumene, isopropyl myristate, lactic acid, laurylamine, mesityl oxide, methoxypropanol, methyl isoamyl ketone, methyl isobutyl ketone, methyl laurate, methyl caprylate, methyl oleate, methylene chloride, m-xylene, n-hexane, n-octylamine, octadecanoic acid, octylamine acetate, oleic acid, oleylamine, o-xylene, phenol, polyethylene glycol alcohol, propionic acid, propyl lactate, propylene carbonate, propylene glycol, propylene glycol methyl ether, p-xylene, toluene, triethyl phosphate, triethylene glycol, xylene sulfonic acid, paraffin, mineral oil, trichloroethylene, perchloroethylene, ethyl acetate, amyl acetate, butyl acetate, propylene glycol methyl ether, diethylene glycol methyl ether, methanol, ethanol, isopropanol, and higher molecular weight alcohols such as amyl alcohol, tetrahydrofurfuryl alcohol, hexanol, octanol, ethylene glycol, propylene glycol, glycerol, N-methyl-2-pyrrolidone and the like.

[0219] Suitable solid carriers are, for example, talc, titanium dioxide, pyrophyllite clay, silica, attapulgite clay, diatomaceous earth, limestone, calcium carbonate, bentonite, calcium montmorillonite, cottonseed hulls, wheat flour, soybean flour, pumice, wood flour, ground walnut shells, lignin, and the like.

[0220] Advantageously, many surface-active substances can be used in both solid and liquid formulations, especially in those formulations that can be diluted with a carrier before use. Surface-active substances can be anionic, cationic, nonionic or polymeric, and they can be used as emulsifiers, wetting agents, or suspending agents, or for other purposes. Typical surface-active substances include, for example, salts of alkyl sulfates, such as diethanolammonium lauryl sulfate; salts of alkylarylsulfonic acids, such as calcium dodecylbenzenesulfonate; alkylphenol / alkylene oxide addition products, such as nonylphenol ethoxylate; alcohol / alkylene oxide addition products, such as tridecyl alcohol ethoxylate; soaps, such as sodium stearate; salts of alkylnaphthalenesulfonic acids, such as sodium dibutylnaphthalenesulfonate; dialkyl esters of sulfosuccinates, such as sodium di(2-ethylhexyl)sulfosuccinate; sorbitan esters, such as sorbitan oleate; quaternary amines, such as lauryltrimethylammonium chloride, polyethylene glycol esters of fatty acids, such as polyethylene glycol stearate; block copolymers of ethylene oxide and propylene oxide; and salts of mono- and di-alkyl phosphates; and further substances described, for example, in McCutcheon's Detergents and Emulsifiers Annual, MC Publishing Corp., Ridgewood NJ (1981).

[0221] Further adjuvants that can be used in the nitrification inhibitor formulations include: crystallization inhibitors, viscosity modifiers, suspending agents, dyes, antioxidants, foaming agents, light absorbers, mixing aids, defoamers, complexing agents, neutralizing or pH modifying substances and buffers, corrosion inhibitors, fragrances, wetting agents, take-up enhancers, trace nutrients, plasticizers, flow aids, lubricants, dispersants, thickeners, antifreeze agents, microbicides, and liquid and solid fertilizers.

[0222] The composition according to the invention may include an additive comprising an oil of plant or animal origin, a mineral oil, an alkyl ester of such an oil, or a mixture of such an oil and an oil derivative. The amount of the oil additive in the composition according to the invention is generally 0.01 to 10%, based on the mixture to be applied. As an example, the oil additive may be added to the spray tank in the desired concentration after the spray (spray) mixture is prepared. Preferred oil additives include mineral oils or oils of plant origin, such as rapeseed oil, olive oil or sunflower oil, emulsified vegetable oils, alkyl esters of oils of plant origin, such as methyl derivatives, or oils of animal origin such as fish oil or tallow. Preferred oil additives include C8-C 22 Alkyl esters of fatty acids, especially C 12 -C 18 Methyl derivatives of fatty acids, such as the methyl esters of lauric acid, palmitic acid, and oleic acid (methyl laurate, methyl palmitate, and methyl oleate, respectively).

[0223] The composition according to the invention generally comprises 0.1 to 99% by weight, in particular 0.1 to 95% by weight, of a compound of the invention, and 1 to 99.9% by weight of a formulation aid, which may include 0 to 25% by weight of a surface-active substance. While commercial products may preferably be formulated as concentrates, end users will generally employ dilute formulations.

[0224] The rate of application varies within wide limits and depends on: the nature of the soil, the method of application, the crop plant, the type of fertilizer used, the prevailing climatic conditions, and other factors controlled by the method of application, the time of application and the target crop. As a general guideline, the compound can be applied at a rate of 1 to 2000 L / ha, especially 10 to 1000 L / ha.

[0225] The composition may further include a urease inhibitor.

[0226] Throughout the specification and appended claims, unless the context indicates otherwise, the word "comprising" will be understood to imply the inclusion of stated integers or steps, or groups of integers or steps, but not the exclusion of any other integers or steps, or groups of integers or steps.

[0227] The reference to any prior publication (or information obtained therefrom), or the reference to any known matter in this specification is not and should not be considered as an acknowledgement or recognition or any form of indication that the prior publication (or information obtained therefrom) or the known matter constitutes part of the common general knowledge in the field to which this specification relates.

[0228] The present invention will now be described with reference to the following non-limiting examples:

[0229] 1. Synthetic nitrification inhibitor

[0230] 1.1 Overall

[0231] The progress of the reaction was monitored by thin layer chromatography (TLC) using silica gel 60 aluminum bottom plates coated with the fluorescent indicator F254 (Merck). The plates were visualized using UV radiation (254 nm) alone or in combination with ninhydrin, potassium permanganate or iodine based stains. Purification by silica gel chromatography was performed using Davisil Chromatographic Silica Media LC60A 40-63 micron with the specified solvent system. All 1 H and 13 C NMR spectra were recorded on a 400 MHz Varian INOVA spectrometer (at 400 or 101 MHz, respectively) downfield from the residual solvent peak using the solvent resonance (solvent peak) as an internal standard (1 H NMR: CDCl3 at 7.26 ppm, DMSO-d6 at 2.50 ppm; 13 C NMR: CDCl3 at 77.0 ppm, DMSO-d6 at 39.5 ppm). Chemical shifts are reported in parts per million (ppm, δ), with the fragmentation pattern indicated as follows: s, singlet; d, doublet; t, triplet; q, quartet; p, quintet; m, multiplet; dd, doublet of doublet. Coupling constants (J) are reported in Hertz (Hz). Electrospray ionization high-resolution mass spectrometry (HRMS) was performed on a Thermo Scientific Exactive Plus Orbitrap spectrometer (Thermo, Bremen, Germany) operated in positive mode.

[0232] 1.2 Overall Procedure A: Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) for the synthesis of 1,4-disubstituted triazoles

[0233]

[0234] Scheme 1: Reaction scheme for general procedure A.

[0235] In the flask under argon atmosphere, sodium azide (1.2 or 1.5 equivalents) is suspended in DMF (0.85M), and suitable alkyl bromide (1 equivalent) is added thereto. The solution is stirred at room temperature for 6-17 hours. The reaction is quenched by adding H o (DMF / H o, 1:1 v / v), and CuSO is added successively afterwards .5H o (0.06 equivalent), sodium ascorbate (0.3 equivalent) and suitable alkynes (1.2 or 1.5 equivalents). Under strong stirring, at 70 ℃, the reaction is heated overnight. The reaction is cooled to room temperature, diluted with water (at least 3x DMF volume) afterwards, and extracted with ethyl acetate. The extract is combined, washed with 5% LiCl aqueous solution and concentrated, and purified by silica chromatography afterwards.

[0236] 1.3 Overall Procedure B: Thermal Huisgen 1,3-Dipolar Cycloaddition for the Synthesis of 1,4,5-Trisubstituted Triazoles

[0237]

[0238] Scheme 2: Reaction scheme for overall procedure B.

[0239] Sodium azide (1.5 eq.) and appropriate alkyl bromide (1 eq.) were filled into a flask purged with argon. They were suspended in DMSO (1.28 M) and warmed to 45 ° C with vigorous stirring. After 20 hours, the reaction was cooled to room temperature and quenched with H2O (DMF / H2O, 4:5 v / v) and then extracted with ether. Under N2 flow, the ether extract was concentrated into oil, which was used directly in subsequent steps. * Note: Organic azides can be explosive, do not evaporate to dryness. Smaller (less) azides are treated with solvent substitution, where toluene is added before evaporating the ether under N2 flow.

[0240] The crude azide is suspended in toluene (0.21 M) and then the appropriate internal alkyne (1.1 equivalents) is added. The reaction is then heated at 115 ° C with vigorous stirring. Once completed (by TLC) (24 to 48 hours), the reaction is cooled. Toluene is removed in vacuo to leave the crude triazole as a waxy brown solid. Purification of the crude product is achieved by recrystallization or column chromatography.

[0241] 1.4 Synthesis of 1-butyl-4-pentyl-1H-1,2,3-triazole (1)

[0242]

[0243] Synthesized by general procedure A; starting from sodium azide (25.6 mmol), 1-bromobutane (17.1 mmol), CuSO4.5H2O (1.0 mmol), sodium ascorbate (5.1 mmol), and 1-heptyne (25.5 mmol), 1 (2.14 g, 11.0 mmol, 64%) was obtained. The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 4:1; R f =0.27).

[0244] Yield: 64% (colorless liquid).

[0245] 1 H NMR (400MHz, CDCl3): δ7.22(s,1H),4.26(t,J=7.3Hz,2H),2.69–2.60(m,2H),1.82(p,J=7.4 Hz, 2H), 1.61 (p, J = 7.4Hz, 2H), 1.37–1.21 (m, 6H), 0.89 (t, J = 7.4Hz, 3H), 0.88–0.80 (m, 3H).

[0246] 13C NMR (101MHz, CDCl3): δ148.30,120.32,49.78,32.27,31.40,29.14,25.61,22.35,19.66,13.93,13.40.

[0247] HRMS(ESI+)m / z:[C 11 H 21 N3+H] + Calculated: 196.18082, measured: 196.18098.

[0248] 1.5 Synthesis of 1,4-butyl-1H-1,2,3-triazole (2)

[0249]

[0250] Synthesized by general procedure A; starting from sodium azide (25.6 mmol), 1-bromobutane (17.1 mmol), CuSO4.5H2O (1.0 mmol), sodium ascorbate (5.1 mmol) and 1-hexyne (25.5 mmol), 2 (2.33 g, 12.9 mmol, 75%) was obtained. The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 4:1; R f =0.19).

[0251] Yield: 75% (colorless liquid).

[0252] 1 H NMR (400MHz, CDCl3): δ7.22(s,1H),4.25(t,J=7.2Hz,2H),2.65(t,J=7.8Hz,2H),1 .81(p,J=7.4Hz,2H), 1.59(p,J=7.6Hz,2H), 1.38–1.23(m,4H), 0.93–0.83(m,6H).

[0253] 13 C NMR (101MHz, CDCl3): δ148.25, 120.33, 49.78, 32.26, 31.55, 25.31, 22.25, 19.65, 13.74, 13.39.

[0254] HRMS(ESI+)m / z:[C 10 H 19 N3+H] + Calculated: 182.16517, measured: 182.16539.

[0255] 1.6 Synthesis of 4-butyl-1H-1,2,3-triazole-1-acetic acid ethyl ester (3)

[0256]

[0257] Synthesized by general procedure A; starting from sodium azide (25.5 mmol), ethyl bromoacetate (17.0 mmol), CuSO4.5H2O (1.0 mmol), sodium ascorbate (6.0 mmol), and 1-hexyne (25.5 mmol), 3 (1.99 g, 9.44 mmol, 56%) was obtained. The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 4:1; R f =0.15).

[0258] Yield: 56% (white solid).

[0259] 1 H NMR (400MHz, CDCl3): δ7.38(s,1H),5.07(s,2H),4.19(q,J=7.1Hz,2H),2.68(t,J=7.7Hz,2H ), 1.61 (p, J = 7.7Hz, 2H), 1.33 (h, J = 7.3Hz, 2H), 1.23 (t, J = 7.6Hz, 3H), 0.87 (t, J = 7.4Hz, 3H).

[0260] 13 C NMR (101MHz, CDCl3): δ166.48,148.71,121.97,62.17,50.69,31.36,25.24,22.17,13.97,13.72.

[0261] HRMS(ESI+)m / z:[C 10 H 17 N3O2+H] + Calculated: 212.13935, measured: 212.13977.

[0262] 1.7 Synthesis of 1-butyl-4-(α,α-dimethylmethanol)-1H-1,2,3-triazole (4)

[0263]

[0264] Synthesized by general procedure A; starting from sodium azide (25.4 mmol), 1-bromobutane (17.0 mmol), CuSO4.5H2O (1.0 mmol), sodium ascorbate (5.8 mmol), and 2-methyl-3-butyn-2-ol (25.5 mmol), 4 (3.12 g, 17.0 mmol, 100%) was obtained. The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 1:1; Rf =0.22).

[0265] Yield: quantitative (yellow liquid).

[0266] 1 H NMR (400MHz, CDCl3): δ7.43 (s, 1H), 4.31 (t, J = 7.3Hz, 2H), 2.77 (s, 1H), 1.87 (p, J = 7.4Hz, 2H), 1.62 (s, 6H), 1.35 (h, J = 7.4Hz, 2H), 0.94 (t, J = 7.4Hz, 3H).

[0267] 13 C NMR (101MHz, CDCl3): δ155.49, 118.90, 68.46, 50.04, 32.24, 30.46, 19.72, 13.43.

[0268] HRMS(ESI+)m / z:[C9H 17 N3O+H] + Calculated: 184.14444, measured: 184.14458.

[0269] 1.8 Synthesis of 4-butyl-1H-1,2,3-triazole-1-butyric acid ethyl ester (5)

[0270]

[0271] Synthesized by general procedure A; starting from sodium azide (8.0 mmol), ethyl 4-bromobutyrate (8.4 mmol), CuSO4.5H2O (0.7 mmol), sodium ascorbate (4 mmol), and 1-hexyne (8.0 mmol), 5 (1.08 g, 4.5 mmol, 56%) was obtained. The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 3:1; R f =0.24).

[0272] Yield: 56% (pale yellow oil).

[0273] 1H NMR (400MHz, CDCl3): δ7.24(s,1H),4.34(t,J=6.9Hz,2H),4.08(q,J=7.1Hz,2H),2.65(t,J=7.7Hz,2H),2.28(t,J=7.1H z, 2H), 2.15 (p, J = 6.9Hz, 2H), 1.59 (p, J = 7.6Hz, 2H), 1.33 (h, J = 7.4Hz, 2H), 1.20 (t, J = 7.1Hz, 3H), 0.87 (t, J = 7.4Hz, 3H).

[0274] 13 C NMR (101MHz, CDCl3): δ172.32,148.43,120.62,60.60,48.95,31.50,30.70,25.46,25.28,22.24,14.12,13.75.

[0275] HRMS(ESI+)m / z:[C 12 H 21 O2N3+H] + Calculated: 240.17065, measured: 240.17061.

[0276] 1.9 Synthesis of substituted triazoles 6-8 via phthalimide-protected intermediates

[0277]

[0278] Scheme 3: Reaction scheme for the synthesis of triazole 6.

[0279] 1.10 Synthesis of 4-butyl-1H-1,2,3-triazole-1-propylamine (6)

[0280]

[0281] Synthesized by a modified reported procedure (Pyta, K. et al., European Journal of Medicinal Chemistry 2014, 84, 651; Wang, Y.-F. et al., Organic Letters 2013, 15 (11), 2842). N-(3-Bromopropyl)phthalimide (10.1 mmol) and sodium azide (15.2 mmol) were dissolved in DMF (26 mL) under argon and stirred at room temperature for 7 hours. The reaction was diluted with H2O (26 mL), followed by the addition of CuSO4.5H2O (1.0 mmol), sodium ascorbate (5.3 mmol), and 1-hexyne (25.5 mmol). The reaction was heated at 70°C with vigorous stirring.

[0282] After 16 hours, the reaction was cooled to room temperature, then diluted with H2O (80 mL) and extracted with ethyl acetate (3 x 80 mL). The extracts were combined, concentrated, and purified by silica chromatography (Pet. ether / EtOAc, 2:3; R f =0.33). If the crude product failed to solidify due to residual DMF, the sample was treated with 5% aqueous LiCl solution to cause precipitation of 2-[3-(4-butyl-1H-1,2,3-triazol-1-yl)propyl]-1H-isoindole-1,3(2H)-dione as a cream powder (84%).

[0283] 1 H NMR (400MHz, CDCl3): δ7.89–7.80(m,2H),7.78–7.68(m,2H),7.44(s,1H),4.36(t,J=6.9Hz,2H),3.74(t,J=6.5Hz,2 H), 2.68 (t, J = 7.7Hz, 2H), 2.31 (p, J = 6.8Hz, 2H), 1.63 (p, J = 7.4Hz, 2H), 1.37 (h, J = 7.4Hz, 2H), 0.92 (t, J = 7.4Hz, 3H).

[0284] 13 C NMR (101MHz, CDCl3): δ168.29,148.41,134.17,131.87,123.36,121.00,47.58,35.11,31.53,29.44,25.33,22.30,13.81.

[0285] HRMS(ESI+)m / z:[C 17 H 20 N4O2+H] + Calculated: 313.16590, measured: 313.16592.

[0286] 2-[3-(4-butyl-1H-1,2,3-triazole-1-1)propyl]-1H-isoindole-1,3(2H)-dione (8.5 mmol) was dissolved in ethanol (0.06 M) and then treated with hydrazine monohydrate (12.67 mmol). The solution was stirred vigorously and heated to 90°C. After heating overnight, a white precipitate formed. The reaction was cooled and the precipitate was removed by filtration and washed thoroughly. The filtrate was concentrated and the resulting solid was resuspended in CH2Cl2 and filtered again. The filtrate was concentrated to a yellow oil, which was purified by silica chromatography (CH2Cl2 / MeOH / 30% NH3 in water, 10:1:0.1; R f=0.16) to give 6 (1.01 g, 5.5 mmol, 65%) as a cream solid.

[0287] Yield: 65% (cream solids).

[0288] 1 H NMR (400MHz, DMSO-d6): δ7.80 (s, 1H), 4.33 (t, J = 7.0Hz, 2H), 2.57 (t, J = 7.6Hz, 2H), 2.47 (t, J = 6. 6Hz, 2H), 1.82 (p, J = 6.8Hz, 2H), 1.64–1.44 (m, 4H), 1.29 (h, J = 7.3Hz, 2H), 0.87 (t, J = 7.4Hz, 3H).

[0289] 13 C NMR (101MHz, DMSO-d6): δ147.17, 122.06, 47.40, 38.88, 34.08, 31.60, 25.14, 22.12, 14.10.

[0290] HRMS(ESI+)m / z:[C9H 18 N4+H] + Calculated: 183.16042, measured: 183.16057.

[0291]

[0292] Scheme 4: Reaction scheme for the synthesis of triazoles 7 and 8.

[0293] 1.11 Synthesis of 2-[3-[4,5-bis(hydroxymethyl)-1H-1,2,3-triazole]propyl]-isoindoline-1,3-dione (7)

[0294]

[0295] Sodium azide (9.4 mmol) was suspended in DMF (26 mL) under argon, and to this solution was added N-(3-bromopropyl)phthalimide (8.7 mmol). The mixture was stirred overnight at room temperature. The reaction was then slowly diluted with H2O (100 mL) and then extracted with ether. Concentration of the ether extract afforded N-(3-azidopropyl)phthalimide (1.83 g, 7.94 mmol, 92%) as a waxy cream solid.

[0296] 1H NMR (400MHz, CDCl3): δ7.90–7.79(m,2H), 7.77–7.67(m,2H), 3.78(t,J=6.8Hz,2H), 3.38(t,J=6.7Hz,2H), 1.96(p,J=6.8Hz,2H).

[0297] 13 C NMR (101MHz, CDCl3): δ168.25, 134.04, 132.00, 123.31, 49.04, 35.38, 28.03.

[0298] HRMS(ESI+)m / z:[C 11 H 10 O2N4+H] + Calculated: 231.08765, measured: 231.08771.

[0299] N-(3-azidopropyl)phthalimide (7.9 mmol) was suspended in toluene (0.2 M) before adding 2-butyne-1,4-diol (8.7 mmol). The reaction was stirred vigorously and heated to 115° C. for 41 hours. Toluene was evaporated and the crude solid was recrystallized from H 2 O to provide 7 (1.34 g, 4.3 mmol, 54%) as a white powder.

[0300] Yield: 54% (white powder).

[0301] 1 H NMR (400MHz, DMSO-d6): δ7.89–7.77(m,4H),5.29(t,J=5.4Hz,1H),5.01(t,J=5.6Hz,1H),4.57(d,J=5 .3Hz, 2H), 4.46 (d, J = 5.5Hz, 2H), 4.37 (t, J = 7.3Hz, 2H), 3.65 (t, J = 7.0Hz, 2H), 2.18 (p, J = 7.2Hz, 2H).

[0302] 13 C NMR (101MHz, DMSO-d6): δ168.36,145.05,134.76,134.52,132.16,123.44,54.63,51.10,45.99,35.69,28.88.

[0303] HRMS(ESI+)m / z:[C 15 H 16 O4N4+H] + Calculated: 317.12443, measured: 317.12448.

[0304] 1.12 Synthesis of 2-[3-[4,5-(methyl acetate)-1H-1,2,3-triazole]propyl]-isoindoline-1,3-dione (8)

[0305]

[0306] For 7, 2-butyne-1,4-diol diacetate was used as the alkyne, and the crude product was recrystallized from ethanol to afford 8 (3.69 g, 9.23 mmol, 71%) as white crystals.

[0307] Yield: 71% (white crystals).

[0308] 1 H NMR (400MHz, CDCl3): δ7.89–7.80(m,2H),7.78–7.69(m,2H),5.23(s,2H),5.22(s,2H),4 .46–4.37(m,2H),3.82(t,J=6.7Hz,2H),2.36(p,J=6.9Hz,2H),2.06(s,3H),2.04(s,3H).

[0309] 13 C NMR (101MHz, CDCl3): δ170.67,170.02,168.20,142.09,134.17,131.89,130.65,123.36,56.67,52.64,46.51,35.23,29.09,20.82,20.50.

[0310] HRMS(ESI+)m / z:[C 19 H 20 N4O6+H] + Calculated: 401.14556, measured: 401.14563.

[0311] 1.13 Synthesis of 4,5-bis(hydroxymethyl)-1H-1,2,3-triazole-1-butyric acid ethyl ester (9)

[0312]

[0313] Synthesized by a modification of the overall procedure B; starting from sodium azide (22 mmol) and ethyl 4-bromobutyrate (19 mmol), heated in DMF (20 mL), 9 (1.21 g, 4.6 mmol, 33%) was obtained. The resulting crude azide was treated with 2-butyne-1,4-diol (14 mmol) in toluene at 115 °C for 24 h. The crude mixture was purified by silica chromatography (CH2Cl2 / CH3OH, 10:0.6; R f =0.28).

[0314] Yield: 33% (pale yellow oil).

[0315] 1 H NMR (400MHz, CDCl3): δ4.94 (s, 2H), 4.67 (s, 2H), 4.58 (s, 2H), 4.37 (t, J = 7.1Hz, 2H), 4. 06(q,J=7.1Hz,2H), 2.32(t,J=7.1Hz,2H), 2.16(p,J=7.1Hz,2H), 1.20(t,J=7.1Hz,3H).

[0316] 13 C NMR (101MHz, CDCl3): δ172.65,144.67,134.23,60.73,55.06,51.82,47.53,30.77,24.99,14.10.

[0317] HRMS(ESI+)m / z:[C 10 H 17 N3O4+H] + Calculated: 244.12918, measured: 244.12921.

[0318] 1.14 Synthesis of 4,5-bis(methyl acetate)-1H-1,2,3-triazole-1-butyric acid ethyl ester (10)

[0319]

[0320] Synthesized by general procedure B; 10 (2.5 g, 7.7 mmol, 74%) was obtained from sodium azide (15.8 mmol), ethyl 4-bromobutyrate (10.5 mmol), and 2-butyne-1,4-diol (11.1 mmol). The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 3:2; R f =0.1).

[0321] Yield: 74% (colorless oil).

[0322] 1H NMR (400MHz, CDCl3): δ5.24(s,2H),5.23(s,2H),4.42(t,J=7.1Hz,2H), 4.11(q,J=7.1Hz,2H), 2.39 (t, J = 7.0 Hz, 2H), 2.22 (p, J = 7.1 Hz, 2H), 2.06 (s, 3H), 2.05 (s, 3H), 1.24 (t, J = 7.1 Hz, 3H).

[0323] 13 C NMR (101MHz, CDCl3): δ172.24,170.65,170.01,142.06,130.64,60.71,56.71,52.69,47.65,30.73,25.15,20.80,20.56,14.15.

[0324] HRMS(ESI+)m / z:[C 14 H 21 N3O6+H] + Calculated: 328.15031, measured: 328.15021.

[0325] 1.15 Synthesis of ethyl 4,5-bis(methyl acetate)-1H-1,2,3-triazole-1-acetate (11)

[0326]

[0327] Synthesized by general procedure B; 11 (1.9 g, 6.3 mmol, 63%) was obtained from sodium azide (15.1 mmol), ethyl 2-bromoacetate (10.0 mmol), and 2-butyne-1,4-diol diacetate (10.7 mmol). The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 1:1:R f =0.43).

[0328] Yield: 63% (pale yellow oil).

[0329] 1 H NMR (400MHz, CDCl3): δ5.25 (s, 2H), 5.24 (s, 2H), 5.23 (s, 2H), 4.24 (q, J = 7.1Hz, 2H), 2.06 (s, 3H), 2.03 (s, 3H), 1.29 (t, J = 7.1Hz, 3H).

[0330] 13C NMR (101MHz, CDCl3): δ170.68,170.16,166.23,142.21,131.79,62.50,56.60,52.97,49.78,20.81,20.48,14.04.

[0331] HRMS(ESI+)m / z:[C 12 H 17 N3O6+H] + Calculated: 300.11901, measured: 300.11890.

[0332] 1.16 Synthesis of ethyl 4,5-bis(hydroxymethyl)-1H-1,2,3-triazole-1-acetate (12)

[0333]

[0334] Synthesized by the reported procedure (Wen, Y.-n. et al., Nucleosides, Nucleotides and Nucleic Acids 2016, 35 (3), 147). 12 (0.85 g, 3.9 mmol, 33%) was obtained from sodium azide (14.3 mmol), ethyl 2-bromoacetate (13.5 mmol) and 2-butyne-1,4-diol (12.1 mmol). The crude mixture was purified by silica chromatography (CH2Cl2 / CH3OH, 10:1; R f =0.13).

[0335] Yield: 33% (white solid).

[0336] 1 H NMR (400MHz, DMSO-d6): δ5.34(t,J=5.5Hz,1H),5.31(s,2H),5.08(t,J=5.7Hz,1H),4.5 7(d,J=5.4Hz,2H), 4.50(d,J=5.5Hz,2H), 4.15(q,J=7.1Hz,2H), 1.20(t,J=7.1Hz,3H).

[0337] 13 C NMR (101MHz, DMSO-d6): δ167.58,144.81,135.15,61.82,54.61,51.67,49.70,14.40.

[0338] HRMS(ESI+)m / z:[C8H 13 N3O4+H] + Calculated: 216.09788, measured: 216.09734.

[0339] 1.17 Synthesis of 1-butyl-4-propyl-1H-1,2,3-triazole (13)

[0340]

[0341] Synthesized by general procedure A; starting from sodium azide (20.3 mmol), 1-bromobutane (17.0 mmol), CuSO4.5H2O (1.0 mmol), sodium ascorbate (5.2 mmol), and 1-pentyne (20.0 mmol), 13 (2.55 g, 15.2 mmol, 89%) was obtained. The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 3:2; R f =0.37).

[0342] Yield: 89% (colorless oil).

[0343] 1 H NMR (400MHz, CDCl3): δ7.23(s,1H),4.28(t,J=7.2Hz,2H),2.66(t,J=7.6Hz,2H),1. 84(p,J=7.3Hz,2H), 1.66(h,J=7.4Hz,2H), 1.32(h,J=7.4Hz,2H), 0.96–0.89(m,6H).

[0344] 13 C NMR (101MHz, CDCl3): δ148.11, 120.38, 49.82, 32.29, 27.66, 22.70, 19.68, 13.74, 13.43.

[0345] HRMS(ESI+)m / z:[C9H 17 N3+H] + Calculated: 168.14952, measured: 168.14951.

[0346] 1.18 Synthesis of 1-(2-methoxyethyl)-4-butyl-1H-1,2,3-triazole (14)

[0347]

[0348] 14 (0.75 g, 4.1 mmol, 65%) was obtained from purified 15 (6.3 mmol) which was dissolved in dry THF (42 mL) under argon and cooled to 0°C. NaH (6.6 mmol) was added in a single portion. Once gas evolution ceased, MeI (9.5 mmol) was added in three portions and the mixture was stirred at room temperature for 24 hours. The reaction was diluted with H2O and THF was removed in vacuo. The product was extracted into ethyl acetate and concentrated. The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 1:1; R f =0.21).

[0349] Yield: 65% (colorless oil).

[0350] 1 H NMR (400MHz, CDCl3): δ7.36(s,1H),4.45(t,J=5.0Hz,2H),3.71(t,J=5.0Hz,2H),3.32(s,3H ), 2.68 (t, J = 7.8Hz, 2H), 1.63 (p, J = 7.8Hz, 2H), 1.36 (h, J = 7.3Hz, 2H), 0.90 (t, J = 7.4Hz, 3H).

[0351] 13 C NMR (101MHz, CDCl3): δ148.26, 121.61, 70.91, 58.93, 50.06, 31.52, 25.31, 22.29, 13.78.

[0352] HRMS(ESI+)m / z:[C9H 17 N3O+H] + Calculated: 184.14444, measured: 184.14445.

[0353] 1.19 Synthesis of 4-propyl-1H-1,2,3-triazole-1-ethanol (15)

[0354]

[0355] Synthesized by general procedure A; starting from sodium azide (26.2 mmol), 2-bromoethanol (17.5 mmol), CuSO4.5H2O (1.0 mmol), sodium ascorbate (5.6 mmol), and 1-hexyne (25.5 mmol), 15 (1.06 g, 6.3 mmol, 36%) was obtained. The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 2:3; R f =0.1).

[0356] Yield: 36% (light yellow liquid).

[0357] 1 H NMR (400MHz, CDCl3): δ7.41(s,1H),4.70(s,1H),4.37(t,J=5.2Hz,2H),3.95(t,J=5.1Hz,2 H), 2.62–2.53 (m, 2H), 1.54 (p, J = 7.4Hz, 2H), 1.29 (h, J = 7.3Hz, 2H), 0.85 (t, J = 7.4Hz, 3H).

[0358] 13 C NMR (101MHz, CDCl3): δ147.87,121.99,60.71,52.62,31.36,25.11,22.20,13.73.

[0359] HRMS(ESI+)m / z:[C8H 15 N3O+H] + Calculated: 170.12879, measured: 170.12887.

[0360] 1.20 Synthesis of 1,4-dipropyl-1H-1,2,3-triazole (16)

[0361]

[0362] Synthesized by general procedure A, starting from sodium azide (20.0 mmol), 1-bromopropane (17.0 mmol), CuSO4.5H2O (1.0 mmol), sodium ascorbate (5.1 mmol), and 1-pentyne (20.1 mmol), 16 (0.68 g, 4.36 mmol, 26%) was obtained. The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 2:1; R f =0.23).

[0363] Yield: 26% (colorless oil).

[0364] 1 H NMR (400MHz, CDCl3): δ7.24 (s, 1H), 4.25 (t, J = 7.2Hz, 2H), 2.66 (t, J = 7.5Hz, 2H), 1.89 (h, J = 7.3Hz, 2H), 1.66 (h, J = 7.4Hz, 2H), 0.96–0.89 (m, 6H).

[0365] 13C NMR (101MHz, CDCl3): δ148.10, 120.46, 51.71, 27.64, 23.72, 22.70, 13.74, 11.04.

[0366] HRMS(ESI+)m / z:[C8H 15 N3+H] + Calculated: 154.13387, measured: 154.13385.

[0367] 1.21 Synthesis of 1-(3-butyn-1-yl)-4-propyl-1H-1,2,3-triazole (17)

[0368]

[0369] Synthesized by a modification of the overall procedure A. 17 (0.30 g, 1.8 mmol, 18%) was obtained from sodium azide (15.0 mmol) and 1-bromobutyne (10.0 mmol) heated at 60 °C in DMF (15 mL) for 3 h. The reaction was cooled and diluted with H2O (15 mL) before adding CuSO4.5H2O (1.2 mmol), sodium ascorbate (2.0 mmol), and 1-pentyne (17.9 mmol). The reaction was stirred overnight at room temperature. The crude mixture was purified by silica chromatography (Pet. ether / diethyl ether, 2:1; R f =0.17).

[0370] Yield: 18% (pale yellow oil).

[0371] 1 H NMR (400MHz, CDCl3): δ7.39(s,1H),4.46(t,J=6.7Hz,2H),2.76(td,J=6.7,2.6Hz,2H),2 .68(t,J=7.6Hz,2H), 2.05(t,J=2.6Hz,1H), 1.68(h,J=7.4Hz,2H), 0.95(t,J=7.4Hz,3H).

[0372] 13 C NMR (101MHz, CDCl3): δ148.14, 121.10, 79.69, 71.32, 48.53, 27.60, 22.66, 20.65, 13.72.

[0373] HRMS(ESI+)m / z:[C9H 13 N3+H] + Calculated: 164.11822, measured: 164.11824.

[0374] 1.22 Synthesis of 1-(2-propen-1-yl)-4-propyl-1H-1,2,3-triazole (18)

[0375]

[0376] Synthesized by a modification of the general procedure A. Sodium azide (21.0 mmol), allyl bromide (15.0 mmol), CuSO4.5H2O (0.9 mmol), sodium ascorbate (4.5 mmol), and 1-pentyne (21.0 mmol) were heated at 45°C overnight to give 18 (1.77 g, 11.7 mmol, 78%). The crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 4:1; R f =0.13).

[0377] Yield: 78% (colorless oil).

[0378] 1 H NMR (400MHz, CDCl3): δ7.26(s,1H),6.08–5.93(m,1H),5.37–5.27(m,1H),5.33–5.20(m,1H),4 .94(dt,J=6.1,1.4Hz,2H), 2.69(t,J=7.6Hz,2H), 1.69(h,J=7.4Hz,2H), 0.96(t,J=7.3Hz,3H).

[0379] 13 C NMR (101MHz, CDCl3): δ148.50, 131.58, 120.39, 119.73, 52.54, 27.69, 22.70, 13.75.

[0380] HRMS(ESI+)m / z:[C8H 13 N3+H] + Calculated: 152.1182, measured: 152.1183.

[0381] 1.23 Synthesis of ethyl 2-(4-propyl-1H-1,2,3-triazol-1-yl)-acetate (19)

[0382]

[0383] Synthesized by general procedure A; sodium azide (20 mmol), ethyl bromoacetate (17.1 mmol), CuSO4.5H2O (1.0 mmol), sodium ascorbate (5.1 mmol), and 1-pentyne (20 mmol), heated at 50°C for 9 hours to obtain 19 (2.83 g, 14.4 mmol, 85%). After workup, the crude mixture was purified by silica chromatography (Pet. ether / EtOAc, 1:1; R f =0.27).

[0384] Yield: 85% (cream waxy solid)

[0385] 1 H NMR (400MHz, CDCl3): δ7.41(s,1H),5.11(s,2H),4.24(q,J=7.2Hz,2H),2.71(t,J =7.6Hz, 2H), 1.70 (h, J = 7.4Hz, 2H), 1.28 (t, J = 7.1Hz, 3H), 0.96 (t, J = 7.4Hz, 3H).

[0386] 13 C NMR (101MHz, CDCl3): δ166.44, 148.62, 122.02, 62.28, 50.77, 27.58, 22.55, 14.03, 13.69.

[0387] HRMS(ESI+)m / z:[C9H 15 N3O2+H] + Calculated: 198.12370, measured: 198.12385.

[0388] 1.24 Synthesis of prop-2-en-1-yl 2-(4-propyl-1H-1,2,3-triazol-1-yl)-acetate (20)

[0389]

[0390] Following the procedure reported by Sabbah et al. (Sabbah, M. et al., Bioorganic & Medicinal Chemistry, 2012, 20 (15), 4727-4736), 19 was saponified into the corresponding acid using ethanol KOH. Under argon in dichloromethane (15 mL), the resulting acid (1.9 mmol), 4-dimethylaminopyridine (0.2 mmol), and allyl alcohol (4.4 mmol) were combined. After stirring for 30 minutes, the solution was cooled to 0 ° C, and then N, N'-dicyclohexylcarbodiimide (2 mmol) was added. The solution was stirred at room temperature for 24 hours, then filtered and concentrated in vacuo. The residue was redispersed in ethyl acetate and filtered again. The filtrate was concentrated into an oil, which was purified by silica chromatography (Pet. ether / EtOAc, 1: 1, R f =0.33).

[0391] Yield: 70% (white waxy solid)

[0392] 1 H NMR (400MHz, CDCl3): δ7.42(s,1H),5.97–5.82(m,1H),5.38–5.24(m,2H),5.15(s,2H), 4.72–4.64(m,2H),2.72(t,J=7.6Hz,2H),1.71(h,J=7.3Hz,2H),0.97(t,J=7.3Hz,3H).

[0393] 13 C NMR (101MHz, CDCl3): δ166.15,148.67,130.83,122.02,119.62,66.68,50.72,27.58,22.56,13.71.

[0394] HRMS(ESI+)m / z:[C 10 H 15 N3O2+H] + Calculated: 210.12370, measured: 210.12392.

[0395] 1.25 Synthesis of prop-2-en-1-yl 2-(4-propyl-1H-1,2,3-triazol-1-yl)-acetamide (21)

[0396]

[0397] Following the procedure reported by Sabbah et al. (Sabbah, M. et al., Bioorganic & Medicinal Chemistry, 2012, 20 (15), 4727-4736), 19 was saponified into the corresponding acid using ethanol KOH. The resulting acid (1.8 mmol) was dissolved in dichloromethane / dimethylformamide (20 mL / 2.5 mL) under argon and treated with HOBt (4.0 mmol), N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (4.0 mmol), and allylamine (4.0 mmol). After stirring at room temperature for 10 hours, the mixture was treated with 5 drops of acetic acid and washed with H2O and brine. The crude product was purified by silica chromatography (CH2Cl2 / MeOH, 100: 1 to 100: 5 gradient, R f =0.1).

[0398] Yield 56% (cream solids).

[0399] 1 H NMR (400MHz, CDCl3): δ7.46(s,1H),6.43(s,1H),5.83–5.69(m,1H),5.14–5.07(m,2H),5.05( s,2H),3.91–3.82(m,2H),2.71(t,J=7.6Hz,2H),1.71(h,J=7.4Hz,2H),0.97(t,J=7.4Hz,3H).

[0400] 13 C NMR (101MHz, CDCl3): δ165.15,148.87,133.01,122.53,116.89,53.07,42.00,27.47,22.49,13.72.

[0401] HRMS(ESI+)m / z:[C 10 H 16 N4O+H] + Calculated: 209.13969, measured: 209.13990.

[0402] 1.26 Synthesis of prop-2-yn-1-yl 2-(4-propyl-1H-1,2,3-triazol-1-yl)-acetate (22)

[0403]

[0404] Following the procedure reported by Sabbah et al. (Sabbah, M. et al., Bioorganic & Medicinal Chemistry, 2012, 20 (15), 4727-4736), 19 was saponified into the corresponding acid using ethanol KOH. The resulting acid (1.8mmol), 4-dimethylaminopyridine (0.4mmol) and propargyl alcohol (3.4mmol) were combined in dichloromethane (15mL) under argon. After stirring for 30 minutes, the solution was cooled to 0 ° C, and N, N'-dicyclohexylcarbodiimide (1.9mmol) was added. The mixture was stirred at room temperature for 24 hours, filtered and concentrated in vacuo. The residue was resuspended in ethyl acetate and filtered again. The filtrate was concentrated into oil, which was purified by silica chromatography (Pet. ether / EtOAc, 1: 1, R f =0.33).

[0405] Yield: 71% (colorless liquid)

[0406] 1 H NMR (400MHz, CDCl3): δ7.43(s,1H),5.19(s,2H),4.79(d,J=2.5Hz,2H),2.72(t,J =7.6Hz, 2H), 2.53 (t, J = 2.4Hz, 1H), 1.71 (h, J = 7.4Hz, 2H), 0.97 (t, J = 7.4Hz, 3H).

[0407] 13 C NMR (101MHz, CDCl3): δ165.77,148.74,122.06,76.30,76.07,53.45,50.52,27.55,22.53,13.71.

[0408] HRMS(ESI+)m / z:[C 10 H 13 N3O2+H] + Calculated: 208.10805, measured: 208.10828.

[0409] 1.27 Synthesis of prop-2-yn-1-yl 2-(4-propyl-1H-1,2,3-triazol-1-yl)-acetamide (23)

[0410]

[0411] Following the procedure reported by Sabbah et al. (Sabbah, M. et al., Bioorganic & Medicinal Chemistry, 2012, 20 (15), 4727-4736), 19 was saponified to the corresponding acid using ethanol KOH. The resulting acid (1.8 mmol) was dissolved in dichloromethane / dimethylformamide (20 mL / 2.5 mL) under argon and treated with HOBt (3.7 mmol), EDCl (4.0 mmol), and allylamine (3.5 mmol). After stirring at room temperature for 10 hours, the mixture was treated with 5 drops of acetic acid and washed with H2O and brine. The crude product was purified by silica chromatography (CH2Cl2 / MeOH, 100:1 to 100:5 gradient, R f =0.1).

[0412] Yield 61% (cream solid)

[0413] 1 H NMR (400MHz, CDCl3): δ7.47(s,1H),6.79–6.71(m,1H),5.06(s,2H),4.04(dd,J=5.4,2.5Hz,2 H), 2.71 (t, J = 7.6Hz, 2H), 2.21 (t, J = 2.6Hz, 1H), 1.71 (h, J = 7.4Hz, 2H), 0.97 (t, J = 7.4Hz, 3H).

[0414] 13 C NMR (101MHz, CDCl3): δ165.04, 148.87, 122.60, 78.45, 72.03, 52.84, 29.38, 27.47, 22.48, 13.73.

[0415] HRMS(ESI+)m / z:[C 10 H 14 N4O+H] + Calculated: 207.12404, measured: 207.12411.

[0416] 2. Soil Experiment

[0417] The soil used in this study was collected from four different locations in Victoria, Australia: (i) wheat-growing soil (36 ° 45 'S, 142 ° 07 'E) from Horsham, (ii) crop rotation (rotational planting) soil (36 ° 37 'S, 142 ° 09 'E) from Dahlen, (iii) vegetable-growing soil (38 ° 08 'S, 145 ° 20 'E) from Clyde, and (iv) pasture soil (38 ° 15 'S, 142 ° 52 'E) from Terang. In addition, the sugarcane-growing soil (17 ° 34 'S, 145 ° 57 'E) from South Johnstone, North Queensland, was also studied. The water content of the soil was measured by oven-drying the sample to a constant weight before starting each experiment. The water-filled pore space (WFPS) of the soil was in the range of 52%-61%, which is within the recommended range of 50-70% for microbial activity due to oxygen and nutrient availability (Fichtner, T. et al., Applied Sciences, 2019, 9, 496).

[0418] The pH values ​​and residual (initial) concentrations of ammonium-N and nitrate-N in the tested soils are compiled in Table 1 below.

[0419] 3,4-Dimethylpyrazole phosphate (DMPP) (prepared as a solution of 3,4-dimethylpyrazole in phosphoric acid) was obtained from Incitec Pivot Fertilisers.

[0420] Table 1. Residual concentrations of ammonium-N and nitrate-N and pH of the soils tested in this study

[0421]

[0422] 2.1 Soil incubation experiment

[0423] Soil microcosm incubations were performed in 250 ml polypropylene sample containers (Sarstedt, Germany) containing 18.24 g oven-dry weight equivalent of soil. Microcosms were rewetted and preincubated for 7 days at the test temperature to allow recovery of soil microbial activity. After preincubation, the remaining volume to achieve the desired water-filled pore space (WFPS%) was applied as one of the following treatment solutions; (NH4)2SO4 (control), (NH4)2SO4+DMPP, or (NH4)2SO4+one of compounds 1-23. For each soil type, each treatment was applied in triplicate, resulting in n=3 at each time point.

[0424] Treatment solutions were prepared so that each small experimental ecosystem received (NH4)2SO4 at a rate of 100 mg N / kg soil, compounds 1-23 at 10 mol % of the applied N, or DMPP (referred to as L-DMPP, M-DMPP, or H-DMPP, respectively) at 1.5, 3.6, or 10 mol % of the applied N.

[0425] The microcosms were incubated for 0, 3, 7, 14, 21, or 28 days, with day 0 samples extracted after 1 hour of incubation following treatment. The soil microcosms were aerated throughout the incubation period, and the moisture content was replenished every few days based on weight loss.

[0426] At the end of each incubation period, the soil microbial ecosystem was destructively sampled by treatment with 2M KCl (100 mL). After shaking for 1 hour, the soil-KCl solution was filtered (Whatman 42), after which the filtrate was stored at -20°C until the end of the experiment. Then, after appropriate dilution of all KCl extracts, nitrogen from ammonium (NH4 + -N) and from NO3 - and NO2 - Nitrogen (NO x - -N) concentrations were analyzed. The results are reported as the mean of three replicates, and the reported errors are the standard error of the mean.

[0427] 2.1.2 Calculation of nitrification percentage and nitrification inhibition percentage

[0428] In the untreated soil NH4 + -N or NO3 - Nitrification calculations were performed as previously reported (Aulakh, M. et al., Biology and Fertility of Soils 2001, 33, 258–263; Mahmood, T. et al., Soil Research 2017, 55, 715-722) with correction for the initial baseline concentration of -N. For each treatment, nitrified NH4 was calculated according to Equation 1 + -N(%):

[0429]

[0430] Among them, [NH4 + -N]0 is the NH4 in the soil at day 0 +-N concentration (in mg N kg -1 Soil), and [NH4 + -N] t is the NH4 in the soil at a given time point t + -N concentration (in mg N kg -1 Soil meter).

[0431] For each treatment, as shown in the following (Equation 2), even during the 28-day incubation experiment, the NO x - -N accumulation rate (mgNO x - -N / kg soil / day):

[0432]

[0433] [NO x - -N] t＝0 and [NO x - -N] t＝28 The nitrite (NO2 - ) and nitrate (NO3 - ) (in mg N kg -1 Soil meter).

[0434] Nitrification inhibition (%) was calculated as follows: Based on NH4 + -N data (i.e., nitrified NH4 calculated by Eq. 1 + -N percentage), or based on NO x - -N data. For NH4 + For the inhibition of nitrification of -N, the percentage value is calculated from the nitrified NH4 of the fertilized control ((NH4)2SO4 only) at a given time point t according to Equation 3 + -N percentage and nitrified NH4 in treated samples ((NH4)2SO4 and NI) at the same time point + -N percentage calculation:

[0435]

[0436] For NO-based x - For nitrification inhibition based on -N concentration, the percentage value is calculated from the NO in the fertilized control ((NH4)2SO4 only) at a given time point t according to Equation 4: x - -N concentration and NO in treated samples ((NH4)2SO4 and NI) at the same time pointx - -N concentration calculation:

[0437]

[0438] 2.1.3 Statistical analysis

[0439] All data presented are the average of three replicates. The raw NH4 + -N and NO x - -N data were statistically analyzed. Data were evaluated for statistical significance (P < .05) by two-way analysis of variance (ANOVA; Chambers & Hastie, 1992), evaluating the effects of the two factors "day" and "treatment", and using Tukey HSD post-hoc adjustment to evaluate pairwise comparisons between treatments at each time point. Statistical results of comparisons of inhibitor treatments with the fertilized control (NH4)2SO4 treatment and DMPP treatment are shown in the original NH4 + -N and NO x - -N data is described in the table.

[0440] 2.1.4NH4 + -N and NO x -N concentration

[0441] Incubation tests comparing compounds to low DMPP (1.5 mol%, L-DMPP) or medium DMPP (3.6 mol%, M-DMPP) treatments were performed in all soils to obtain initial structure-activity relationship information to guide future synthesis. Selected compounds were retested against high DMPP (10 mol%, H-DMPP) treatments in alkaline soils (Horsham, Darren) along with compounds 13 to 17. In most studies, fertiliser NH4 applied in the control (NH4)2SO4 treatments within 28 days + -N is completely consumed. In summary, in most alkaline soils (Horsham, Darren), the nitrification inhibitors are effective in slowing down NO x -N is the most efficient in terms of generation.

[0442] Trom soil

[0443] In acidic Trom soil, compound 3 and L-DMPP maintained more NH4 in the soil than the fertilized control. +-N was the most effective treatment (see Table 2 below). Reduced NO was also observed for these treatments when compared to the fertilized control. x - -N was produced, predominantly until day 14, after which the concentration converged to that of the control.

[0444] Table 2. Nitrified ammonia (%) during 28 days of incubation at 25°C in Trom soil (pH 6.5). Bold values ​​indicate lower nitrification rates than those observed in the control treatment ((NH4)2SO4), positively correlated with inhibitor activity. Errors stated are standard errors of the mean, n=3.

[0445]

[0446] Horsham Soil

[0447] Depend on Figure 1 As is clear from Table 3, among the various inhibitors tested at 25°C, compounds 13, 16, 17 (and to a lesser extent compound 2) performed statistically better in terms of maintaining NH4 + (where P < .001 for these compounds, except for compound 2), and inhibition of NO x - Formation (P = .013 (13), .001 (16), < .001 (17)). At 35 ° C, all compounds 2, 13, 16, and 17 showed nitrification rates lower than the uninhibited control treatment. Among these, compound 13 and DMPP performed statistically better in maintaining NH4 + (P = .004 (13) and .008 (DMPP)), and preventing NO x - The results were (P = .03 for both treatments), with compound 13 being slightly more effective than DMPP in delaying the nitrification of ammonia (61% vs. 65% NH4 + -N consumption).

[0448] Table 3. Ammonia nitrified (%) during 28 days incubation at 25°C or 35°C in Horsham soil (pH 8.8). Bold values ​​indicate nitrification rates lower than those observed in the control treatment ((NH4)2SO4), positively correlated with inhibitor activity. Errors stated are standard errors of the mean, n=3.

[0449]

[0450]

[0451] Figure 2The calculated NO shown in x - -N production rates showed that in all treatments, incubation at 25 °C resulted in lower NO compared to those at 35 °C. x - -N accumulation, except for compounds 2 and 14, in which NO x - -N accumulation was lower at elevated temperatures. NO in soil treated with compound 13 x - The rate of -N accumulation was the same at both temperatures (2.8 mg NO x - -N / kg soil / day), while the treatment with H-DMPP showed the greatest increase in production rate at the higher tested temperatures.

[0452] Darren Soil

[0453] In Dahlen soil, all compounds 2, 13, 16, and 17 showed significant increases in the NH4 + -N was statistically superior (P < .001). The results from these tests are shown in Figure 3 and Table 4. At an elevated temperature of 35°C, all four triazoles outperformed H-DMPP in slowing the rate of ammonia nitrification. Figure 3 NO shown in B and 3D x - The considerable error in the -N measurements may be due to the fact that before starting the test, the soil was mixed with other soils (Horsham NO3 - -N: 7.2mg kg -1 ; Trump NO3 - -N: 27mg kg -1 ) compared to the soil, this soil is particularly rich in NO3 - (NO3 - -N:270mg kg -1 ).

[0454] Incubation studies in this soil at 35°C using DMPP and compounds 2, 13, 16, and 17 (data included in Table 4) revealed that DMPP performed significantly worse at higher temperatures, while all compounds 2, 13, 16, and 17 maintained NH4 after 28 days compared to both the DMPP treatment and the control treatment. + Statistically better performance was observed in all 14 samples (P < .001), with ammonia consumption ranging from 17% (16) to 38% (17). + -N and NO x- -N concentration is shown in Figure 3 C and 3D.

[0455] NO in soil during the 28-day incubation period x - The rate of -N accumulation is shown in Figure 4 Thus, for all treatments, incubations at 25°C resulted in higher NO x - -N accumulation, except for DMPP. Treatment with 16 at 35 °C resulted in the lowest accumulation rate (1.8 mg NO x - -N / kg soil / day), while for the treatment with compound 17 at 25°C, the highest accumulation rate (4.7 mg NO x - -N / kg soil / day). Interestingly, for compound 17 at 35 °C, the accumulation rate was reduced to 2.4 mg NO x - -N / kg soil / day, which is the largest reduction in accumulation rate for all inhibitors tested in this series. On the other hand, NO in soil treated with compound 13 x - The rate of -N accumulation was minimally affected by temperature changes (2.5 vs. 2.4 mg NO at 25°C and 35°C, respectively). x - -N / kg soil / day), very similar to the seemingly temperature-independent behavior observed for this compound in the Horsham soil.

[0456] Table 4. Ammonia nitrified (%) during 28 days of incubation at 25°C or 35°C in Dahlen soil (pH 7.3). Bold values ​​indicate nitrification rates lower than those observed in the control treatment ((NH4)2SO4), positively correlated with inhibitor activity. Errors stated are standard errors of the mean, n=3.

[0457]

[0458] Further comparative tests were carried out in Dahlen soil for compounds 18, 20 and 23 against H-DMPP at both 25°C and 35°C. The results of these tests are shown in Figure 5 and Table 5 below. Again, it should be noted that Figure 5 B and 5D show that for NO x - The considerable error in the N measurements may be due to the fact that this soil is particularly rich in NO3 -(See above).

[0459] Table 5. Ammonia nitrified (%) during 28 days incubation at 25°C or 35°C in Dahlen soil (pH 7.3). Bold values ​​indicate nitrification rates lower than those observed in the control treatment ((NH4)2SO4), positively correlated with inhibitor activity. Errors stated are standard errors of the mean, n=3.

[0460]

[0461]

[0462] At 25°C, by day 28, compounds 18 and 23 were significantly more effective than both the fertilized control and DMPP in maintaining NH4 + -N levels were statistically better (P < .001). Compound 20 performed statistically better than the fertilized control (P < .001). However, this effectiveness was not reflected in the NO x - -N concentration, none of the treatments showed significant effectiveness compared to the control treatment or DMPP, respectively, on day 28.

[0463] At 35 °C, compared with those observed at 25 °C, the + -N and NO x - The trends in N concentrations were less linear, especially for NO x - By day 28, compounds 18 and 20 and DMPP were the only ones that maintained NH4 in the soil compared to the (NH4)2SO4 treated control. + -N remained highly effective (18: P < .01, 20: P < .001). However, treatments with compounds 18 and 20 did not reflect the NH4 observed for DMPP treatment at day 21. + -N concentration, which corresponds to a considerable loss of inhibitory activity (52% nitrified ammonia compared to only 41% for the control, see Table 5). x - In terms of NO data, all NIs (except DMPP) showed lower NO compared to the control at day 28. x - -N concentration, however, did not reach statistical significance.

[0464] South Johnston Soil

[0465] Compared to the other soils studied, the detected NH4 + -N and NO x - -N concentration behaved differently. The increase in NH4 observed during the first 14 days + -N concentrations indicate that mineralization of nitrogen is occurring in the soil. Tests at 35°C included a water-only control treatment in addition to the (NH4)2SO4 treated control, which shows that mineralization is occurring in the soil regardless of treatment and is stimulated by the addition of nitrogen-based fertilizers. This "complexity" makes it more difficult to assess the impact of the tested compounds on the nitrification process, as NH4 + -N starts to show a net loss rather than a net gain (from mineralization). Table 6 shows the amount of ammonia lost compared to the amount detected for the selected treatments at day 0 for both the tests at 25°C and 35°C, while Figure 6 Explanation of the measured NH4 + -N and NO x - -N amount. For almost all entries, the percentage is negative, indicating that the [NH4 + -N] remained above the percentage detected on day 0 (due to the mineralization process). From day 14 onwards, the more negative percentages indicated that the treatment was effective in preventing [NH4 + -N] loss is more effective.

[0466] At 25°C, all treatments maintained NH4 at 21 and 28 days compared with the (NH4)2SO4 control treatment. + -N and mitigation of NO x - -N growth is significantly better (see Figure 6 Of these treatments, compound 19 and DMPP were the least effective (although still significantly better than the control treatment). Compounds 3 and 18 both showed significantly higher effectiveness compared to the treatment with DMPP, based on higher NH4 + -N and lower NO x - -N concentration (3: P < .05 and < .01, respectively; 18: P < .01 and < .001, respectively).

[0467] Table 6. Ammonia nitrified (%) during a 28-day incubation in South Johnston soil (pH 5.0). Nitrification values ​​were calculated from the ammonia levels detected in samples at each time point. All samples were expressed as 100 mg N kg -1The ratio of the fertilizer (NH4)2SO4 treatment was used. The bold values ​​indicate nitrification rates that are lower than those observed in the control treatment ((NH4)2SO4), positively correlated with inhibitor activity. The values ​​are reported as means (n=3); the errors reported are the standard errors of the means.

[0468]

[0469] At 35°C, only a subset of compounds 3, 16, and 18 maintained NH4 + -N and mitigation of NO x - -N growth was statistically superior (P < .001). At this temperature, compounds 3, 16, and 18 also maintained NH4 + -N had statistically better performance (P < .001 for 3 and 16, P < .05 for 18).

[0470] 2.2. Leaching study of DMP and 16

[0471] The leachability (solubility) of soil nitrification inhibitors is an important consideration because of the potential cascading health consequences if chemical inhibitors migrate through the soil profile and enter groundwater supplies at high concentrations. It is also an important consideration for the effectiveness of the inhibitors because high mobility in soil can lead to the accumulation of inhibitors, NH4 + Spatial separation between ions and microorganisms involved in the nitrification process results in reduced field effectiveness.

[0472] Traditional soil leaching (leaching) columns are material-intensive and time-intensive and may not be used due to limited access to the soil of interest. Therefore, soil thin layer chromatography (TLC) techniques were developed by modifying methods previously described for studying the leaching behavior of pesticides (insecticides) (Helling, CS, Turner, BC, Science 1968, 162, 562-563). The advantage of the TLC technique is that it provides data very quickly and requires only small amounts of soil and matrix.

[0473] 2.2.1. Overall soil TLC plate preparation

[0474] TLC plates are prepared based on the method described in the literature (Helling, CS, Turner, BC, Science 1968, 162, 562-563; Mohammad, A., Jabeen, N., JPC-Journal of Planar Chromatography-Modern TLC 2003, 16, 137-143). 3 columns (4 cm W x 12 cm H) are outlined on a glass TLC plate (20x20 cm) using masking tape (3 layers, ~450 μm total thickness). A slurry of freshly ground soil in distilled H2O (~2:3 m / v) is then poured onto the prepared plate and evenly spread using a glass rod. Once uniform, the plate is dried overnight in an oven at 35°C. Careful removal of the masking tape provides a TLC plate ready for sample application.

[0475] 2.2.2. Leaching studies of DMP and compound 16

[0476] Samples of DMP (the active core of DMPP) or compound 16 (~1 mg) dissolved in acetone (100 μL) were pipetted across the soil in a straight line, 2 cm above the base of the plate. The applied tape thickness was kept below 0.5 cm. After a drying time of 30 minutes, the TLC plate was developed in a glass developing chamber with distilled HO (depth 0.5 cm) until the solvent front reached the top of the soil (~1 hour). If the solvent front failed to move through three adjacent soil channels at the same rate, the plate was removed as soon as the solvent reached the top of one channel, and any dry soil in the remaining channel was carefully scraped off to mark the height of the solvent front. The plate was then allowed to air dry overnight.

[0477] Once dry, the board is divided into 6 horizontal bands corresponding to the following R f Values: (1) <0.05 (baseline), (2) 0.05 to 0.25, (3) 0.25 to 0.45, (4) 0.45 to 0.65, (5) 0.65 to 0.85, and (6) 0.85 to 1. In sequence, the soil in each band was carefully hung off the glass backing and collected in a vial. Special care was taken to avoid cross contamination between soils from different bands and separate channels.

[0478] 2.2.3. Extraction and analysis of DMP and compound 16

[0479] Individual soil bands collected from the TLC plates were extracted as follows:

[0480] 1. Add an aqueous solution of CaCl2 / MgSO4 (0.01 M and 0.45 M, respectively, 2 mL).

[0481] 2. Sonicate for 5 minutes, then shake manually for 30 seconds.

[0482] 3. Add methyl tert-butyl ether (MTBE, 2 mL) and shake manually for 30 seconds.

[0483] 4. Sonicate for 10 minutes, then shake manually for 30 seconds.

[0484] 5. Leave it alone until the soil begins to settle, then freeze it at -20°C overnight.

[0485] 6. After partial thawing, the ether extract was filtered through a nylon syringe filter ( 13 mm diameter, 0.45 μm pore size) filtration.

[0486] The filtered ether extract (450 μL) was spiked with 50 μL of a standard solution of cyclododecanone in MTBE (1.97 mg / mL for samples treated with DMP) or a standard solution of cyclooctanone in MTBE (6.37 mg / mL for samples treated with 16). The samples were then directly analyzed by GC-MS (method: 70°C, 5 min, then 10°C / min ramp to 250°C, hold at 250°C for 17 min [total run time = 40 min]) to analyze the presence or absence of inhibitors in each soil zone.

[0487] 2.2.4 Calculation of leached inhibitors

[0488] For standard cyclooctanone (R t = 9.1 min) and cyclododecanone (R t =15.8 min) were compared with the GC-MS peak areas calculated for compound 16 (R t =13.9 minutes) and DMP (R t =7.6 minutes) for each soil sample extract in which the inhibitor was detected as follows:

[0489]

[0490] Then for samples from a single TLC channel;

[0491]

[0492] Because each treatment was run in triplicate, f Mean values ​​are reported as percentages of inhibitor detected, with errors presented as standard deviations.

[0493] Table 7. Results from soil thin layer chromatography (TLC) for evaluating the leaching potential of inhibitor DMP and compound 16 in two soils.

[0494]

[0495]

[0496] a The mean values ​​were obtained from three replicates, and the errors presented are standard deviations. b Values ​​were calculated compared to the internal standard cyclooctanone. c Values ​​are calculated compared to the internal standard cyclododecanone. Not detected = nd

[0497] 2.2.5. DCD leaching study

[0498] A sample of DCD (~1 mg) dissolved in methanol (300 μL) was pipetted across the soil in a straight line, 2 cm above the base of the plate. The applied tape thickness was maintained at 0.5 cm. After 30 minutes of drying time, the TLC plate was developed in distilled H2O (depth 0.5 cm) in a glass development chamber until the solvent front reached the top of the soil (~1 hour). If the solvent front failed to move the three adjacent soil channels of the lattice at the same rate, the plate was removed once the solvent reached the top of the soil in one channel, and any dry soil in the remaining channels was carefully scraped off to mark the height of the solvent front. Subsequently, the plate was allowed to air dry overnight.

[0499] Once dry, the board is divided into 6 horizontal bands corresponding to the following R f Values: (1) <0.05 (baseline), (2) 0.05 to 0.25, (3) 0.25 to 0.45, (4) 0.45 to 0.65, (5) 0.65 to 0.85, and (6) 0.85 to 1. In sequence, the soil in each band was carefully scraped off the glass backing and collected in a vial. Special care was taken to avoid cross contamination between soils from different bands and separate channels.

[0500] 2.2.6. DCD extraction and analysis

[0501] Individual soil bands collected from the TLC plates were extracted as follows:

[0502] 1. Add methanol (2 mL).

[0503] 2. Manually shake for 30 seconds after 15 minutes of sonication.

[0504] 3. Once the soil begins to settle, filter the methanol extract through a nylon syringe ( 13 mm diameter, 0.45 μm pore size) filtration.

[0505] 4. 300 μL of the methanol extract was evaporated under a stream of nitrogen and the residue was taken up in ultrapure acetonitrile (1 mL).

[0506] 5. Filter the acetonitrile solution through a PVDF syringe ( 33 mm diameter, 0.22 μm pore size) filtration.

[0507] The filtered acetonitrile extracts (10 μL injection) were then directly analyzed by HPLC (1260 Infinity II Preparative LC system with a C18 column, Agilent) to detect the presence or absence of DCD in each soil zone at 214 nm. The HPLC method used was as follows:

[0508] -Solvent A: 0.1% formic acid in H2O

[0509] -Solvent B: 0.1% formic acid in acetonitrile

[0510] - Ramp from 100% A to 100% B over 10 minutes. Hold at 100% B for 2 minutes, then return to 100% A within 10 seconds, with a 2 minute wash at 100% A. Total sample run time = 15 minutes.

[0511] 2.2.7. Results

[0512] Retention factor (R f ) is used to measure the movement of compounds through soil using TLC methods, where a high R close to 1 f The values ​​indicate high mobility through soil. In neutral Dahlen soil, compound 16 showed reduced mobility compared to DMP, with most of the triazole in R f The range of 0.25-0.45 was detected, compared with 0.65-0.85 for DMP (see Figure 7 ). For the acidic South Johnston soil, DMP was found to be leached in a narrower band and to a lesser extent than compound 16. This may be due to the protonation of DMP in a lower pH environment. The resulting charged molecule may adsorb on soil particles, thus reducing leaching. However, because DMP was not the target of this study, the underlying processes were not explored.

[0513] Dicyandiamide (DCD) is a widely used nitrification inhibitor with known leaching concerns due to its high water solubility. Preliminary results from TLC leaching studies of DCD in both Darren and South Johnston soils showed that the greatest DCD accumulation was in the R f This result contradicts the correlation between ease of protonation and reduced mobility, since DCD has multiple protonation sites and is therefore expected to leach less.

[0514] The results from these leaching studies indicate that in neutral soils, compound 16, and by extension other similar small lipophilic triazoles, have lower leachability than DMP and DCD. Again, acidic soils appear to be potentially more problematic, however compound 16 still appears to show lower to similar leaching trends than DCD.

Claims

1. A method for reducing nitrification in soil, comprising treating the soil with a compound of formula (I) or an agriculturally acceptable salt thereof: in R 1 Selected from -C1-C 10 Alkyl; -C1-C4 substituted by one or more groups selected from C1-C4 alkoxy 10 Alkyl; -C2-C 10 Alkenyl; -C2-C 10 Alkynyl; -C1-C 10 Alkyl C(O)OR 4 ;-C1-C 10 AlkylOC(O)R 4 ; and -C1-C 10 Alkyl C(O)NHR 4 ; R 2 Selected from -C1-C 10 Alkyl and -C1-C 10 alkyl; R 3 is H; and R 4 Selected from: -C1-C4 alkyl, -C2-C4 alkenyl, and -C2-C4 alkynyl.

2. The method according to claim 1, wherein the soil is co-treated with a urease inhibitor.

3. A method according to claim 1 or claim 2, wherein the soil is co-treated with a fertilizer.

4. A composition for reducing nitrification in soil comprising a compound of formula (I) as defined in claim 1, and at least one agriculturally acceptable adjuvant or diluent.

5. The composition according to claim 4, further comprising a urease inhibitor.

6. Fertilizer comprising a urea or ammonium based fertilizer and a compound of formula (I) as defined in claim 1.

7. The fertilizer according to claim 6, wherein the urea or ammonium based fertilizer is in the form of granules and the compound of formula (I) is coated on the granules.

8. The fertilizer according to claim 6, further comprising a urease inhibitor.

9. The fertilizer according to claim 8, wherein the urea or ammonium based fertilizer is in the form of granules, and the compound of formula (I) and the urease inhibitor are coated on the granules.

10. Use of a compound of formula (I) as defined in claim 1 for the manufacture of a composition for reducing nitrification in soil, or for the manufacture of a fertilizer.

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