Patent Application
20150119251
The present invention relates to an agricultural plant-protecting agent including a dipeptide derivative or an agro-pharmaceutically acceptable salt thereof as an active ingredient, which has a plant disease-preventing effect, a plant growth-promoting effect, and a plant immunity-activating effect.
The plants make signaling molecules in order to defend themselves when they are invaded by pathogens or physically wounded. It is known that such signaling molecules has the effects as plant defense activators, plant strengthening agents, or plant immunity activators. As the representative signaling molecules of the plants, salicylic acid and jasmonic acid have been known. When the defense reaction of plant is operated due to outside invasions, salicylic acid, a signaling molecule, induces the genes, such as, PR-1, BGL-2, PR-5, SID-2, EDS-5, and PAD-4 to be expressed in the plant, and jasmonic acid induces the genes, such as, PDF1.2, VSP, HEL, THI-2, FAD3, ERS1, and ERF1 to be expressed in the plant [Dong, X., Current Opinion in Plant Biology, 1998, 1, 316-323; Glazebrook, J., Current Opinion in Plant Biology, 1999, 2, 280-286; Bostock, R. M., Physiology and Molecular Plant Pathology, 1999, 55, 99-109]. Particularly, the PR-1 gene is a gene indicating the resistance-inducing phenomenon of a plant, which is generated by salicylic acid, the expression of the PR-1 gene is a crucial sign of operation of the signaling process by a plant. When a PR protein is expressed in a plant, the plant shows a disease resistant effect such as an antibacterial activity. In addition, a PDF1.2 gene is a gene indicating the resistance-inducing phenomenon of a plant, which is induced by Jasmonic acid, the expression of the PDF1.2 gene is a crucial sign of the progress of the signaling process for operation of the defense reaction of plant.
Benzo-1,2,3-thiadiazole-7-carbothioic acid S-ethyl ester (BTH) has been commercially used as a signaling molecule of a plant. BTH is a synthetic compound having the similar chemical structure as salicylic acid. It is reported that BTH shows an activity of suppressing Blumeria graminis that causes a barley powdery mildew, the ozone resistance of a plant, the resistance to Botrytis cinerea in a grape plant, a promoting activity of the biosynthesis of resveratrol and anthocyanin, the resistance to a powdery mildew in a strawberry, and also an accumulating effect of phenolic compounds. BTH has a function of inducing the expression of the PR-1 gene like salicylic acid does, but does not induce the expression of PDF1.2 gene exhibited by jasmonic acid.
As described above, when the signaling molecules such as salicylic acid and jasmonic acid for the self-defense of a plant induce to express the genes such as PR-1 or PDF1.2, the plant can suppress the propagation of the pathogens and also can endure a physical wound. In other words, it is considered that self-defense ability is induced in a plant. Therefore, even if the conventional antimicrobial agent or germicide is not treated, when such genes are expressed in a plant, the plant can resist the pathogen. Therefore, it is considered that the molecule expressing such genes has a great value as an agricultural drug for a plant.
Meanwhile, a dipeptide is a peptide molecule having an amide bond generated by condensing two amino acids. The dipeptide is generated in various types of structures even at the time of performing the proteolysis, and also is generated even during the process of synthesizing 2,5-diketopiperazine, a cyclic dipeptide. Aspartame, that is, an artificial sweetener, belongs to a dipeptide being composed of two amino acids. As being reported so far, a dipeptide compound is mainly applied as an artificial sweetener, a platelet aggregation inhibitor, and the like [International Patent Publication No. WO9616981, U.S. Pat. Nos. 5,968,581 and 5,763,408, and Europe Patent Publication No. 513675].
The present inventors synthesized the dipeptide derivatives prepared by two amino acids, and found that when these compounds are treated to a plant, they express the plant disease-resisting genes such as PR-1, glucanase, chitinase, PR4, peroxidase, and PR10. In addition, we found that when the dipeptide derivatives are treated to a plant before forming the lesion, the formation of the lesion in a plant is significantly suppressed, and also in addition to the plant disease-preventing effect, a plant growth-promoting effect and a plant immunity-increasing effect (in detail, a plant disease-resisting ability or plant cooling-preventing ability) are also improved. As a result, the present inventors completed the present invention.
In other words, the present inventors found at first time that the dipeptide derivatives induce the disease resistance gene to be expressed, and shows a plant disease-preventing effect by suppressing the proliferation of pathogen, a plant growth-promoting effect, and a plant immunity-increasing, and up to now, no case has been reported in any literature.
An object of the present invention is to provide an use of a dipeptide derivative as an agricultural plant-protecting agent.
In order to achieve such an object, the present invention provides an agricultural plant-protecting agent including a compound selected from a dipeptide derivative represented by the following chemical formula 1 or an agro-pharmaceutically acceptable salt thereof as an active ingredient.
In the above chemical formula 1,
R1, R2, and R5 are the same or different from each other, and represent a hydrogen atom, a linear or branched C1-C18 alkyl carbonyl group, a linear or branched C1-C18 alkoxy carbonyl group,
R3, R4, R6, and R7 are the same or different from each other, and represent a hydrogen atom, or a linear or branched C1-C18 alkyl group substituted or unsubstituted with the group selected from hydroxy, mercapto, amino, guanidino, N,N-bis(benzyloxycarbonyl)guanidino, carbamoyl, carboxylic acid, linear or branched C1-C18 alkoxy carbonyl, linear or branched C1-C18 alkenyl oxycarbonyl,
linear or branched C1-C18 alkyl thio, trityl thio, acetyl amino, phenyl, hydroxyphenyl, imidazole, and indolyl;
or any one of the R3 and R4 may bind to R2 to form a nitrogen-containing 5-membered to 7-membered ring or any one of the R6 and R7 may bind to R5 to form a nitrogen-containing 5-membered to 7-membered ring.
R8 represents hydroxy; a linear or branched C1-C18 alkoxy group; a linear or branched C1-C18 alkyl amino group;
Ra represents a linear or branched C1-C18 alkyl group;
n represents an integer of 0 to 5, the number of a substituent, Ra; and
m represents an integer of 0 to 6.
In addition, the present invention provides a method for preventing or suppressing a plant disease, in which the method includes treating an agricultural plant-protecting agent including a compound selected from a dipeptide derivative represented by the above chemical formula 1 or an agro-pharmaceutically acceptable salt thereof as an active ingredient to a plant.
In addition, the present invention provides a method for activating plant immunity, in which the method includes treating an agricultural plant-protecting agent including a compound selected from a dipeptide derivative represented by the above chemical formula 1 or an agro-pharmaceutically acceptable salt thereof as an active ingredient to a plant.
In addition, the present invention provides a method for promoting plant growth, in which the method includes treating an agricultural plant-protecting agent including a compound selected from a dipeptide derivative represented by the above chemical formula 1 or an agro-pharmaceutically acceptable salt thereof as an active ingredient to a plant.
The agricultural plant-protecting agent of the present invention has an effect on significantly reducing the formation of lesion in a plant due to the suppressions of infection and proliferation of pathogens by inducing the production of the plant disease-resisting protein by expressing the plant disease-resisting genes such as PR-1, glucanase, chitinase, PR4, peroxidase, and PR10, when being treated to a plant. In addition, the agricultural plant-protecting agent of the present invention allows a plant to activate the disease resistant mechanism to pathogen or protect itself from chilling damages by operating self-defense mechanism. In addition, the agricultural plant-protecting agent of the present invention allows a plant to tolerate environmental stress caused by cold or hot weather condition, and also has an effect on promoting plant growth.
In addition, the agricultural plant-protecting agent of the present invention has a systemic effect on other regions which was not treated, by the self-defense mechanism of a plant. In other words, even though the agent is treated to a part of a plant, such as seeds, roots, stems, and leaves of a plant, it has an effect on other untreated regions, and even on harvested fruits.
Since the agricultural plant-protecting agent has an excellent effect on preventing plant diseases such as bacterial rot, damping-off, phytophthora blight, a wilt disease, a leaf spot disease or a mosaic disease, which are caused by a bacteria, a virus, or a mold, it is useful as an environment-friendly agricultural plant-protecting agent capable of replacing the conventional germicides and being used.
The agricultural plant-protecting agent of the present invention has an excellent effect in the case of applying to dicotyledoneae, for example, solanaceae such as potatoes, peppers, sweet peppers, and tomatoes, cucurbitaceae, such as cucumbers, cigarettes, watermelons, and oriental melons, a crucifer (cruciferae) such as Chinese cabbage, lettuces, radishes, cabbages, rapes, peanuts, and a salary, a medicinal plant, such as ginseng and dong quai, a plant, such as perilla seeds, strawberry, spring onion, garlic, ginger, and onion, graminae, such as, rice, barley, corn, and sorghum, and fruit trees, such as apple tree, pear tree, peach tree, and persimmon tree.
The present invention relates to an agricultural plant-protecting agent including a dipeptide derivative represented by the above-described chemical formula 1 or an agro-pharmaceutically acceptable salt thereof as an active ingredient.
For the dipeptide derivative represented by the above-described chemical formula 1 which is included, as an active ingredient, in the agricultural plant-protecting agent, in the case of having one or more chiral carbons, the agricultural plant-protecting agent of the present invention may include the compound represented by the above-described chemical formula 1 in a type of a racemic mixture or isomeric compound as an active ingredient.
In the present invention, examples of the agro-pharmaceutically acceptable salt may include a metallic salt, a salt with an organic base, a salt with an inorganic acid, a salt with an organic acid, a salt with a basic or acidic amino acid, and the like. Examples of the suitable metallic salt may include an alkali metallic salt, such as, a sodium salt and a potassium salt; an alkali earth metallic salt, such as, a calcium salt, a magnesium salt, and a barium salt; an aluminum salt; and the like. Examples of the salt with an organic base may include the salts with trimethyl amine, triethyl amine, pyridine, picoline, 2,6-lutidine, ethanol amine, diethanol amine, triethanol amine, cyclohexyl amine, dicyclohexyl amine, N,N-dibenzyl ethylene amine, and the like. Examples of the salt with an inorganic acid may include the salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like. Examples of the salt with an organic acid may include the salt with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like. Examples of the salt with a basic amino acid may include the salts with alginin, lysine, ornithine, and the like. Examples of the salt with an acidic amino acid may include the salts with aspartic acid, glutamic acid, and the like.
The substituent that is used for defining the dipeptide derivative represented by the above-described chemical formula 1 according to the present invention will be described in more detail as follows. The term “alkyl” in the present invention means the linear or branched alkyl group having 1 to 18 carbon atoms, preferably 1 to 6 carbon atoms, and more preferably 1 to 4 carbon atoms. In detail, it may include methyl, ethyl, n-propyl, isopropyl, n-butyl, 1-methyl propyl, 2-methyl propyl, t-butyl, n-pentyl, t-pentyl, n-hexyl, isohexyl, heptyl, octyl, hexadecanyl, octadecanyl, and the like. The term “alkoxy” in the present invention means O-alkyl, and at this time, the alkyl is the same as described above.
In addition, for the dipeptide derivative represented by the above-described Chemical Formula 1, preferably, the R1, R2, and R5 are the same or different from each other, and represent a hydrogen atom, an acetyl group, a hexanoyl group, a hexadecanoyl group, an octadecanoyl group, a benzoyl group, a 4-hexylbenzoyl group, a 2-phenylacetyl group, a 3-phenylpropanonyl group, a methoxycarbonyl group, an ethoxycarbonyl group, a t-butoxycarbonyl group, a hexadecanoxy carbonyl group, an octadecanoxycarbonyl group, a phenoxycarbonyl group, and a 4-hexylbenzyloxycarbonyl group; the R3, R4, R6, and R7 are the same or different from each other, and represent a hydrogen atom, a methyl group, an ethyl group, a n-propyl group, an isopropyl group, a 1-methylpropyl group, a 2-methylpropyl group, a hydroxymethyl group, a 1-hydroxyethyl group, a 2-hydroxyethyl group, an imidazole-4-yl-methyl group, a 2-methylthioethyl group, a benzyl group, a 4-hydroxybenzyl group, a phenethyl group, a mercaptomethyl group, a methylthiomethyl group, a methylthioethyl group, a tritylthiomethyl group, a tritylthioethyl group, a methoxycarbonylmethyl group, an ethoxycarbonylmethyl group, a propanoxycarbonylmethyl group, a tert-butoxycarbonylmethyl group, a pentaneoxycarbonylmethyl group, a hexaneoxycarbonylmethyl group, an allyloxycarbonylmethyl group, a 2-allyloxycarbonylethyl group, a benzyloxycarbonylmethyl group, a benzyloxycarbonylethyl group, a penethyloxycarbonylmethyl group, a 2-penethyloxycarbonylethyl group, a 3-phenylpropyloxycarbonylmethyl group, a 2-(3-phenylpropyloxycarbonyl)ethyl group, a 1-methoxycarbonylethyl group, a 2-methoxycarbonylethyl group, a 2-ethoxycarbonylethyl group, a 2-propaneoxycarbonylethyl group, a 2-butoxycarbonylethyl group, a 2-pentaneoxycarbonylethyl group, a 2-hexaneoxycarbonylethyl group, a 2-aminoethyl group, a carbamoylmethyl group, an acetylaminomethyl group, an acetylaminoethyl group, a carboxymethyl group, a carboxyethyl group, an imidazole-4-ylmethyl group, an imidazole-4-ylethyl group, a 3-guanidinopropyl, a N, N-bis(benzyloxycarbonyl)guanidinopropyl, an indole-3-ylmethyl group, or an indole-3-ylethyl group; or any one of the R3 and R4 may bind to R2 to form a nitrogen-containing 5-membered ring or any one of the R6 and R7 may bind to R5 to form a nitrogen-containing 5-membered ring; and the R8 represents the compound representing a hydrogen atom, a methoxy group, an ethyoxy group, a propaneoxy group, a t-butoxy group, a hexaneoxy group, a hexadecaneoxy group, an octadecaneoxy group, a benzyloxy group, a penethyloxy group, a 3-phenylpropaneoxy group, a methylamino group, an ethylamino group, a propylamino group, a butylamino group, a hexylamino group, a phenylamino group, a benzylamino group, a hexadecylamino group, and an octadecylamino group.
In addition, the dipeptide derivative represented by the above-described chemical formula 1 is the same in more detail as follows:
The dipeptide derivative represented by the above-described chemical formula 1 is an intermediate for synthesizing a 2,5-diketopiperazine compound, and the method for preparing the same is already known in many literatures [Akiyama et al, J. Chem. Soc., Perkin Trans. 1 1989, 235; Gordon et al, Bioorg. Med. Chem. Lett., 1995, 5, 47; Carlsson, A. C. Tetrahedron Lett. 2006, 47, 5199; Lopez-Cobenas, A. Synlett. 2005, 1158; Boehm et al, J. Org. Chem. 1986, 51, 2307; Thajudeen et al, Tetrahedron Lett., 2010, 51, 1303]. Therefore, the person who skilled in the organic synthesis field can easily synthesize the dipeptide derivative represented by the above-described chemical formula 1 with reference to the known literatures.
The dipeptide derivative represented by the above-described chemical formula 1 or agro-pharmaceutically acceptable salt thereof can exhibit a resisting effect on various plant diseases caused by bacteria, virus, and molds by inducing the production of plant disease-preventing proteins through expressing the resistance genes in a plant. Therefore, it is useful as an active ingredient of the agricultural plant-protecting agent. In detail, the plant diseases caused by bacteria, virus, and molds may include a bacterial rot, damping-off, phytophthora blight, a wilt disease, a leaf spot disease or a mosaic disease of a plant. The plants, to which the agricultural plant-protecting agent including the dipeptide derivative represented by the above-described chemical formula 1 as an active ingredient is applied, may be dicotyledoneae, for example, solanaceae such as potatoes, peppers, sweet peppers, and tomatoes, cucurbitaceae, such as cucumbers, cigarettes, watermelons, and oriental melons, a crucifer (cruciferae) such as Chinese cabbage, lettuces, radishes, cabbages, rapes, peanuts, and a salary, a medicinal plant, such as ginseng and dong quai, a plant, such as perilla seeds, strawberry, spring onion, garlic, ginger, and onion, graminae, such as, rice, barley, corn, and sorghum, and fruit trees, such as apple tree, pear tree, peach tree, and persimmon tree. It is preferable to apply to dicotyledoneae, such as, cigarettes, Chinese cabbage, peppers, strawberry, cucumbers, potatoes, and tomatoes.
The agricultural plant-protecting agent of the present invention may include, as an active ingredient, singly, the dipeptide derivative represented by the above-described chemical formula 1 or an agro-pharmaceutically acceptable salt. In addition, the agricultural plant-protecting agent of the present invention includes 0.001 to wt %, and preferably 0.005 to 30 wt % of the active ingredient, and may further include excipient, residually. The excipient may be a general microorganism formulation, an antibacterial effect-promoting agent, diluents, or carrier. In addition to the agricultural active ingredient of the present invention, other germicides, insecticides, herbicides, plant growth-controlling agents or fertilizers, which are conventionally commercialized and used, or developed for increasing a remedial effect or expanding the range of application, are further included in the range of certain content, and thus, may be formulated in a type of mixed formulation.
The excipient and dilutent included in the agricultural plant-protecting agent of the present invention are generally used in the agricultural field, and may be included by further adding solid carriers, for example, oxides, such as, diatomite and slaked lime, phosphate, such as, apatite, a sulfate, such as, gypsum, and mineral powders, such as, clay, kaolin, bentonite, acid clay, quartz, and silica, and filler, anti-aggregating agents, a surfactant, an emulsifying agent, and preservatives thereto. In addition, the agricultural drugs of the present invention may be formulated in order to release the active ingredient in the types of prompt release, sustained release, and delayed release by using the method that are known in the art. For the formulation, additives, such as, the surfactant, diluent, dispersant, and adjuvants that are generally used are mixed with the active ingredient, and then, the mixture thus obtained may be formulated in various types, such as, water dispersible powders, suspension, emulsion, emulsifiable concentrate, microemulsion, liquid formulation, dispersible formulation, particulate water dispersible powders, granule, powdered medicine, liquid water dispersible powder, particulate water dispersible powder, water floating granule, and tablets, and then used.
The agricultural plant-protecting agent of the present invention may be applied to a plant by using the general method. For applying it to a plant, it may be directly scattered or applied to leaves, stems, branches, roots, and seeds of plant body; may be mixed with the general cultivation soils, such as, a paddy field or dry field or nursery bed soil or media; or, in the case of the plant body cultured in water, may be treated to the surface of water in order to prevent the damage of disease. As a specific applying method, there may be an application treatment, a dipping treatment, a fumigation treatment, or a scattering treatment. For example, the agricultural plant-protecting agent may be scattered to soils, or the leaves, stems, seeds, flowers, or fruits of a plant. In order to apply the agricultural plant-protecting agent of the present invention to a plant, the agent may be diluted with water or proper medium, and then used.
In order to confirm the plant disease inducible resistance (ISR) effect, the plant growth-promoting effect, and cold weather-overcoming effect, multidirectional experiments were performed.
First, a dipeptide derivative of the present invention was treated to a pepper plant, and then the degrees of the expressions of the main defense genes, for example, antibiotic genes, such as, PR-1, beta-1,3-glucanase, chitinase, PR4, peroxidase, and PR10 primer were measured. It was confirmed that for the test groups treated with the dipeptide derivative represented by the above-described chemical formula 1, the expressions of the genes suppressing the plant disease outbreak were significantly increased as compared with the control group. In addition, it was also confirmed that the defense genes were not expressed by treatment only with the agents before being inoculated with the plant pathogen, but the resistance mechanism started to operate in the plant infected with the plant pathogen, such as, the pathogen of a bacterial rot (Pectobacterium carotovorum SCC1) and the pathogen of phytophthora (Phytophthora capcisi) (see
In addition, it could be confirmed that when the agricultural plant-protecting agent of the present invention was treated to the plant, the plant growth was promoted up to the maximum 24.4% as compared with the non-treated group (see
As described above, the agricultural plant-protecting agent of the present invention can induce the disease resistance of the plant; also, can exhibit the effect on promoting the growth; and can exhibit the effect to endure the physical stress such as the cold-weather damage. Therefore, the agent of the present invention has very high value for being applied as the next generation agricultural plant-protecting agent having the action mechanism that is completely different from the conventional plant-protecting agents. The plant disease-preventing effect, plant growth-promoting effect, and plant immunity-activating effect (plant disease-resisting effect and cold-weather damage-preventing effect) by the agricultural plant-protecting agent of the present invention will be described in more detail in the following Examples.
As described above, the present invention will be described in more detail with reference to the following Synthesis Examples and Examples, but the present invention is not limited thereto.
(1) Dipeptide Synthesis Method A (Compounds 1, 3, 5, 7, 9, 12 to 24, 26, 27, 29, 32 to 36, 38 to 47, 49, 51 to 60, 62 to 73, 75, 76, 79 to 87, and 106)
Various L- or D-type N-Boc-amino acids (3.0 mmol) were dissolved in 10 mL of dimethylformamide (DMF). Diisopropylmethylamide (DIEA; 0.78 g, 6.0 mmol), 0-(benzotriazole-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (HBTU′ 1.37 g, 3.6 mmol) was added thereto, and then stirred for about 30 minutes. L- or D-type amino acid alkyl (methyl or ethyl) ester (hydrochloride) (3.6 mmol) was added to the reaction mixture thus obtained, and then stirred at room temperature for 12 hours. The reaction mixture thus obtained was diluted with ethyl acetate, and then washed with sodium hydrogen carbonate or salt water. Water was removed from the organic layer with anhydride sodium sulfate, and then the organic layer without water was concentrated under reduced pressure. The concentrated solution thus obtained was purified through a silica column chromatograph (EtOAc/Hexane) to obtain linear N-Boc-L-amino acid-L-amino acid alkyl ester. The yield rate of the condensation reaction was about 50% to 95%.
(2) Dipeptide Synthesis Method B (Compounds 25, 28, 30, 31, 37, 48, 50, 61, 74, 77, 90, 92, and 94)
By using 10.0 mmol of the cysteine, in which a SH group of the cysteine is protected with a trityl group, the aspartic acid or glutamic acid derivative, in which its terminal COOH group is converted with alkyl ester, and lysine or arginine, in which its terminal amino group is protected with a Boc or benzyloxycarbonyl group, the linear N-Boc-L-amino acid-L-amino acid alkyl esters were synthesized by using the same method as Synthesis method A described above. The yield rate of the condensation reaction was about 65% to 80%.
(3) Dipeptide Synthesis Method C (Compounds 2, 4, 6, 8, 10, 11, 78, 88, 89, 91, 93, 95, and 107 to 109)
The N-Boc-amino acid-amino acid alkyl esters (5 mmol) obtained from Synthesis method A and B were dissolved in 50% trifluoroacetic acid-methylene chloride solution (20 mL), and then stirred for 2 hours. The solvent was evaporated under reduced pressure to obtain the amino acid-amino acid alkyl ester trifluoroacetic acid salt without a protecting group, such as, Boc or trityl. As another method, N-Boc-amino acid-amino acid methyl ester (5 mmol) was dissolved in a formic acid (10 mL), stirred for 6 hours, and then vacuum-concentrated to obtain the amino acid-amino acid alkyl ester formic acid without a protecting group. Compound 89 was purchased from Sigma-Aldrich and then used. Compounds 91, 93, and 95 could be obtained from Synthesis method described above, and also could be obtained from Compound 89 by reacting with methyl iodide, allyl bromide, and benzyl chloride, respectively, in MeCN/MeOH.
(4) Dipeptide Synthesis Method D (Compound 11)
The N-Boc-amino acid-amino acid alkyl esters obtained from Synthesis method A and B were again dissolved in water-soluble dioxane, the equivalent LiOH was added thereto, and then the mixture thus obtained was stirred overnight. Since then, the mixture thus obtained was neutralized with acetic acid and was extracted three times with the mixed solvent of acetone/ethanol to obtain the amino acid-amino acid without an alkyl group.
(5) Dipeptide Synthesis Method E (Compounds 96 to 105)
The amino acid-amino acid alkyl ester salt (0.57 mmol) obtained from Synthesis method C was dissolved in methylene chloride (5 mL), acyl chloride (1.2 eq) and triethyl amine (2.4 eq) were added thereto, and then the reaction mixture thus obtained was stirred at room temperature for 2 hours. The reaction mixture thus obtained was added to EtOAc (50 mL), washed with 1 M HCl solution, sodium carbonate, and saturated salt water, and then concentrated. The concentrated solution thus obtained was purified through a silica column chromatograph (EtOAc/Hexane) to obtain N-acyl amino acid-amino acid alkyl ester in the yield rate of 90% or more.
The compounds prepared by using the methods of the representative Synthesis Examples are listed in the following Table 1.
Compound 1
Colorless liquid; 1H NMR (400 MHz, CD3OD) δ 4.17 (1H, t, J=4 Hz), 4.50 (1H, dd, J=6.0, 9.0 Hz), 3.75 (2H, dd, J=11.2, 5.2 Hz), 3.70 (3H, s), 1.63 (1H, m), 1.70-1.62 (2H, m), 1.45 (9H, br s), 0.93 (3H, d, J=6.8 Hz), 0.92 (3H, d, J=6.8 Hz).
Compound 2
Colorless liquid; 1H NMR (400 MHz, CD3OD) δ 4.51 (1H, m), 4.50 (1H, m), 3.87 (2H, d, J=11.2, 5.2 Hz), 3.70 (3H, s), 1.66-1.63 (2H, m), 1.62 (1H, m), 0.95 (3H, d, J=6.4 Hz), 0.92 (3H, d, J=6.4 Hz).
Compound 3
White crystal; 1H NMR (400 MHz, CD3OD) δ 4.12 (1H, m), 3.97 (1H, d, J=17.6 Hz), 3.88 (1H, d, J=17.6 Hz), 3.70 (3H, s), 1.71 (2H, m), 1.53 (1H, m), 1.44 (9H, br s), 0.95 (3H, d, J=6.8 Hz), 0.93 (3H, d, J=6.8 Hz); 13C NMR (100 MHz, CD3OD) δ 174.9, 170.1, 156.4, 79.1, 52.9, 51.1, 40.8, 40.4, 27.2, 24.4, 22.0, 20.4.
Compound 4
1H NMR (400 MHz, CD3OD) δ 4.07 (2H, d, J=17.6 Hz), 3.90 (1H, m), 3.72 (3H, s), 1.63 (1H, m), 1.60-1.70 (2H, m), 0.93 (3H, d, J=6.4 Hz), 0.90 (3H, d, J=6.4 Hz).
Compound 5
White solid; 1H NMR (400 MHz, CD3OD) δ 0.93 (3H, d, J=6.8 Hz), 0.97 (3H, d, J=7.2 Hz), 1.44 (9H, br s), 2.06 (1H, m), 3.67 (3H, s), 3.91 (1H, m), 3.97 (2H, m); 13C NMR (100 MHz, CD3OD) δ 173.6, 170.0, 156.5, 79.1, 59.9, 51.1, 40.3, 30.6, 27.2, 18.2, 16.8.
Compound 6
White solid; 1H NMR (400 MHz, CD3OD) δ 1.06 (3H, d, J=2.8 Hz), 1.08 (3H, d, J=2.8 Hz), 2.20 (1H, m), 3.71 (3H, s), 3.74 (1H, d, J=5.6 Hz), 3.93 (1H, d, J=17.6 Hz), 4.09 (1H, d, J=17.6 Hz); 13C NMR (100 MHz, CD3OD) δ 171.5, 170.3, 59.8, 52.8, 41.9, 31.6, 18.7, 18.0.
Compound 7
Colorless liquid; 1H NMR (400 MHz, CD3OD) δ 4.50 (1H, dd, J=6.0, 5.6 Hz), 4.05 (1H, m), 4.00 (1H, m), 3.69 (3H, s), 1.70 (1H, m), 1.68 (1H, m), 1.60 (1H, m), 1.44 (9H, br s), 1.19 (3H, d, J=6.4 Hz), 0.94 (3H, d, J=6.4 Hz), 0.91 (3H, d, J=6.4 Hz); 3C NMR (100 MHz, CD3OD) δ 173.0, 172.0, 156.5, 79.3, 67.1, 59.9, 51.2, 50.6, 40.1, 27.2, 24.3, 21.8, 20.4, 18.6.
Compound 8
White solid; 1H NMR (400 MHz, CD3OD) δ 4.47 (1H, dd, J=6.0, 6.0 Hz), 4.35 (1H, d, J=6 Hz), 4.10 (1H, q. J=6 Hz), 3.70 (3H, s), 1.70 (1H, m), 1.68 (1H, m), 1.60 (1H, m), 1.21 (3H, d, J=6.4 Hz), 0.95 (3H, d, J=6.8 Hz), 0.90 (3H, d, J=6.8 Hz).
Compound 9
White solid; 1H NMR (400 MHz, CD3OD) δ 4.38 (1H, q, J=7.6 Hz), 4.16 (2H, q, J=7.2 Hz), 4.11 (1H, t, J=4.4 Hz), 1.73 (2H, m), 1.52 (1H, m), 1.43 (9H, br s), 1.38 (3H, d, J=7.2 Hz), 1.25 (3H, t, J=7.2 Hz), 0.95 (3H, d, J=6.8 Hz), 0.93 (3H, d, J=6.8 Hz).
Compound 10
White solid; 1H NMR (400 MHz, CD3OD) δ 4.38 (1H, q, J=7.0 Hz), 4.16 (2H, q, J=7.2 Hz), 3.36 (1H, t, J=6.4 Hz), 1.73 (2H, m), 1.52 (1H, m), 1.38 (3H, d, J=7.2 Hz), 1.25 (3H, t, J=7.2 Hz), 0.96 (3H, d, J=6.4 Hz), 0.93 (3H, d, J=6.4 Hz); 13C NMR (100 MHz, CD3OD) δ 176.3, 172.6, 160.8, 52.7, 48.2, 44.2, 24.2, 21.8, 21.2, 16.0, 13.0.
Compound 11
White solid; 1H NMR (400 MHz, CD3OD) δ 1.84 (1H, m), 1.61 (1H, dd, J=8.4, 5.6 Hz), 1.64 (1H, dd, J=8.4, 5.2 Hz), 3.93 (1H, ddd, J=8.4, 4.4, 0.8 Hz), 3.99 (1H, ddd, J=14.0, 6.8, 0.8 Hz), 1.44 (3H, d, J=6.8 Hz), 0.96 (3H, t, J=6.8 Hz), 0.96 (3H, t, J=6.8 Hz).
Compound 12
White solid; 1H NMR (CDCl3, 400 MHz) δ 8.10 (1H, s), 7.96 (1H, s), 4.60 (1H, q), 4.52 (1H, d), 3.67 (3H, s), 2.66 (1H, m), 1.48 (3H, s), 1.40 (9H, s), 1.01 (6H, d); 13C NMR (CDCl3, 100 MHz) δ 171.6, 171.1, 156.0, 79.5, 60.3, 51.9, 48.2, 31.0, 28.5, 17.1.
Compound 13
White solid; 1H NMR (CDCl3, 400 MHz) δ 6.68 (1H, s), 5.12 (1H, s), 4.59-4.53 (1H, m), 4.17 (1H, s), 3.73 (3H, s), 1.43 (9H, s), 1.37-1.31 (6H, m); 13C NMR (CDCl3, 100 MHz) δ 173.1, 172.3, 155.4, 80.0, 52.4, 49.9, 47.9, 28.2, 18.3, 18.2.
Compound 14
Viscous liquid; 1H NMR (400 MHz, CDCl3): δ 6.50 (1H, s), 6.22 (1H, s), 4.74-4.69 (1H, m), 3.59 (3H, s), 3.49-3.47 (2H, d), 1.54 (9H, s), 1.29-1.27 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 171.6, 170.7, 156.3, 79.5, 51.9, 47.9, 45.0, 28.5, 17.2.
Compound 15
Viscous liquid; 1H NMR (400 MHz, CDCl3) δ 7.02 (2H, d), 6.74 (2H, d), 6.55 (1H, d), 5.11 (1H, s), 4.52 (1H, t), 4.29 (1H, s), 3.70 (3H, s), 2.97 (2H, dd), 1.41 (9H, s), 1.33 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 171.6, 170.7, 156.3, 79.5, 51.9, 47.9, 45.0, 28.5, 17.2.
Compound 16
White solid; 1H NMR (CDCl3, 400 MHz) δ 8.34 (1H, s), 7.50 (1H, d), 7.34 (1H, d), 7.19 (1H, t), 7.12 (1H, t), 6.97 (1H, d), 6.63 (1H, d), 5.11 (1H, m), 4.93 (1H, m), 3.73 (2H, s), 3.66 (3H, s), 3.31 (2H, d), 1.41 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.1, 169.1, 156.2, 136.1, 127.5, 123.0, 122.2, 119.6, 118.3, 111.3, 109.6, 80.2, 52.8, 52.4, 44.2, 38.6, 28.2, 27.5.
Compound 17
White solid; 1H NMR (DMSO-d6, 400 MHz) δ 8.95 (1H, s), 7.50-7.48 (1H, d), 7.16-7.02 (4H, m), 6.72 (1H, s), 6.02 (1H, s), 4.88-4.84 (1H, m), 4.28-4.23 (1H, m), 3.54 (3H, s), 3.25-3.23 (2H, d), 1.55 (9H, s), 1.52-1.47 (1H, m), 1.01-0.99 (6H, d); 13C NMR (DMSO-d6, 400 MHz) δ 175.8, 171.2, 152.5, 135.9, 128.5, 122.8, 120.7, 119.6, 118.1, 112.2, 109.3, 79.9, 55.6, 51.9, 37.8, 28.7 (3C), 26.6, 24.2 (2C), 21.4.
Compound 18
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 7.2 (1H, s), 5.29 (1H, s), 4.45 (2H, d), 3.85 (2H, d), 3.67 (3H, s), 1.45 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 171.5, 171.0, 155.5, 80.3, 51.2, 48.5, 46.1, 29.0.
Compound 19 1H NMR (CDCl3, 400 MHz) δ 7.31-7.20 (5H, m), 5.24 (1H, d), 4.67 (1H, dd), 4.50 (1H, dd), 3.74 (3H, s), 3.61-3.57 (1H, m), 3.19-3.15 (1H, m), 3.11 (1H, dd), 2.93 (1H, dd), 2.17-2.13 (1H, m), 1.96-1.88 (3H, m), 1.37 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.5, 170.8, 155.3, 136.5, 129.9, 128.5, 126.9, 79.8, 60.6, 59.1, 53.4, 52.4, 47.0, 39.4, 29.2, 28.5, 25.0.
Compound 20
Viscous liquid; 1H NMR (CD3OD, 400 MHz) δ 4.46 (1H, dd, J=4.8, 8.8 Hz), 4.37 (1H, q, J=6.8 Hz), 3.77 (1H, m), 3.69 (3H, s), 3.65-3.62 (1H, m), 3.60-3.54 (1H, m), 2.30-2.23 (1H, m), 2.07-1.93 (3H, m), 1.42 (9H, s), 1.28 (3H, d, J=7.6 Hz); 13C NMR (CDCl3, 100 MHz) δ 172.7, 172.6, 156.2, 79.0, 58.9, 51.3, 47.7, 46.6, 28.5, 27.3, 24.5, 15.7.
Compound 21
1H NMR (CDCl3, 400 MHz) δ 5.19 (1H, d), 4.52 (1H, dd), 4.28 (1H, dd), 3.77-3.73 (1H, m), 3.70 (3H, s), 3.64-3.62 (1H, m), 3.59-3.56 (1H, m), 2.23-2.19 (1H, m), 2.03-1.94 (3H, m), 1.40 (9H, s), 1.02 (3H, d), 0.92 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.6, 171.4, 156.0, 79.6, 58.9, 57.0, 52.3, 47.3, 31.5, 29.2, 28.5, 25.2, 19.4, 17.5.
Compound 22
1H NMR (CDCl3, 400 MHz) δ 5.09 (1H, d), 4.45 (1H, dd), 4.39 (1H, dd), 3.71-3.68 (1H, m), 3.62 (3H, s), 3.53-3.51 (1H, m), 2.16-2.11 (1H, m), 2.01-1.86 (3H, m), 1.72-1.65 (1H. m), 1.42 (1H, t), 1.33 (9H, s), 0.92 (3H, d), 0.87 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.6, 171.9, 155.8, 79.5, 58.8, 52.3, 50.4, 46.8, 42.0, 29.1, 28.4, 25.0, 24.6, 23.5, 21.9.
Compound 23
White solid; 1H NMR (CDCl3, 400 MHz) δ 6.50 (1H, s), 5.95 (1H, s), 5.57 (1H, d), 4.47-4.42 (2H, m), 3.66-3.63 (2H, t), 3.64 (3H, s), 2.27-2.22 (2H, m), 2.16-2.04 (2H, m), 1.97-1.89 (3H, m), 1.81-1.77 (1H, m), 1.35 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 175.1, 172.5, 170.6, 155.8, 79.7, 58.7, 52.2, 50.9, 46.9, 31.2, 29.0, 28.9, 28.3, 28.2, 24.9.
Compound 24
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.48 (1H, d), 4.50 (1H, dd), 4.38 (1H, dd), 4.14 (1H, m), 3.76-3.74 (1H, m), 3.69 (3H, s), 3.47 (1H, s), 2.22-2.20 (1H, m), 2.01-1.93 (3H, m), 1.40 (9H, s), 1.19 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.5, 170.9, 156.1, 79.9, 67.4, 58.8, 55.7, 52.7, 52.4, 47.2, 38.5, 31.3, 28.9, 24.8, 18.5.
Compound 25
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.32 (1H, d), 4.81 (1H, dd), 4.46 (1H, dd), 3.73-3.69 (2H, m), 3.67 (3H, s), 2.65 (1H, dd), 2.45 (1H, dd), 2.17-2.12 (1H, m), 2.01-1.90 (3H, m), 1.39 (9H, s), 1.35 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.4, 172.2, 170.0, 155.0, 80.9, 79.7, 59.3, 58.8, 53.4, 52.5, 52.1, 49.1, 48.9, 46.8, 46.5, 31.2, 28.9, 27.9, 24.7.
Compound 26
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 6.85 (1H, s), 6.17 (1H, d), 6.02 (1H, s), 4.78 (1H, dd), 4.42 (1H, dd), 3.76-3.70 (1H, m), 3.61 (3H, s), 2.64-2.46 (1H, m), 2.17-2.12 (1H, m), 2.16-2.04 (2H, m), 1.99-1.89 (3H, m), 1.34 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.8, 172.4, 171.0, 155.5, 79.7, 59.1, 52.5, 52.2, 49.4, 47.1, 46.6, 37.8, 28.9, 28.3, 24.8.
Compound 27
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 4.50 (1H, dd), 4.43 (1H, dd), 3.69 (3H, s), 3.57-3.28 (4H, m), 2.12-1.73 (8H, m), 1.37 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.7, 171.6, 157.7, 79.8, 60.5, 58.7, 51.9, 47.1, 46.0, 29.7, 28.5, 22.6, 22.1.
Compound 28
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.30 (1H, d), 4.83 (1H, s), 4.54 (1H, dd), 4.46 (1H, dd), 3.72 (3H, s), 3.63-3.58 (1H, m), 3.13-3.08 (2H, m), 2.24-2.18 (2H, m), 2.07-1.93 (3H, m), 1.76-1.73 (1H, m), 1.60-1.37 (5H, m), 1.42 (18H, s); 13C NMR (CDCl3, 100 MHz) δ 172.5, 171.6, 156.0, 155.2, 79.6, 58.7, 52.3, 51.5, 46.9, 32.3, 28.9, 28.4, 28.3, 24.9, 21.9.
Compound 29
White solid; 1H NMR (CDCl3, 400 MHz) δ 7.67 (1H, s), 7.42 (1H, s), 6.90 (1H, s), 5.59 (1H, d), 4.59 (1H, dd), 4.48 (1H, dd), 3.71 (3H, s), 3.62-3.58 (1H, m), 3.27-3.22 (1H, m), 3.10-2.97 (2H, m), 2.19 (1H, dd), 1.98-1.82 (3H, m), 1.35 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 173.4, 170.6, 155.2, 135.2, 127.9, 122.5, 79.9, 59.0, 52.6, 51.9, 47.0, 29.5, 28.9, 28.8, 28.3, 28.2, 25.1.
Compound 30
White solid; 1H NMR (CDCl3, 400 MHz) δ 5.34 (1H, d), 4.54-4.49 (1H, m), 4.39 (1H, dd), 3.69 (3H, s), 3.66-3.54 (2H, m), 2.34-2.18 (2H, m), 2.17-1.87 (5H, m), 1.76-1.67 (1H, m), 1.41 (18H, s); 13C NMR (CDCl3, 100 MHz) δ 172.3, 172.1, 170.5, 155.5, 80.4, 79.5, 59.2, 52.6, 52.2, 51.3, 51.0, 49.1, 48.9, 46.9, 46.4, 31.4, 30.6, 28.0, 27.8, 24.7.
Compound 31
Light yellow solid; 1H NMR (CDCl3, 400 MHz) δ 9.38 (1H, s), 9.18 (1H, s), 7.29-7.16 (10H, m), 5.32 (1H, d), 5.14 (2H, s), 5.03 (2H, s), 4.38 (1H, dd), 4.34 (1H, dd), 3.92 (2H, m), 3.56-3.52 (1H, m), 3.50 (3H, s), 3.42-3.38 (1H, m), 2.67 (2H, d), 2.06-2.00 (1H, m), 1.82-1.74 (2H, m), 1.39-1.21 (3H, m), 1.31 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.1, 170.8, 165.5, 163.7, 160.5, 160.4, 155.7, 155.7, 155.4, 137.0, 134.8, 128.6, 128.6, 128.3, 127.9, 127.8, 127.8, 127.6, 79.3, 68.7, 66.8, 58.7, 51.9, 51.5, 46.7, 44.3, 38.5, 29.7, 28.8, 28.3, 24.8, 24.5
Compound 32
White solid; 1H NMR (CDCl3, 400 MHz) δ 7.57 (1H, s), 7.05 (2H, d), 6.70 (2H, d), 5.30 (1H, d), 4.61 (1H, dd), 4.49 (1H, dd), 3.68 (3H, s), 3.61-3.59 (1H, m), 3.28-3.26 (1H, m), 3.00 (1H, dd), 2.82 (1H, dd), 2.13-2.09 (1H, m), 1.95-1.88 (3H, m), 1.35 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.5, 171.2, 155.8, 130.9, 127.1, 115.6, 80.2, 60.6, 59.2, 53.5, 52.4, 47.1, 38.0, 29.1, 28.5, 25.0, 21.2, 14.3.
Compound 33
White solid; 1H NMR (CDCl3, 400 MHz) δ 4.44 (1H, dd), 4.39 (1H, dd), 3.69 (3H, s), 3.61-3.33 (4H, m), 2.21-1.78 (8H, m), 1.43 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 173.2, 172.6, 153.9, 79.9, 59.5, 58.0, 52.3, 52.0, 47.1, 46.0, 30.0, 29.4, 28.4, 24.5.
Compound 34
White solid; 1H NMR (CDCl3, 400 MHz) δ 5.16 (1H, d), 4.42 (1H, dd), 4.34 (1H, dd), 3.90-3.86 (1H, m), 3.70 (3H, s), 3.57-3.51 (1H, m), 2.20-2.17 (1H, m), 2.04-1.92 (3H, m), 1.71-1.66 (1H, m), 1.59-1.53 (1H, m), 1.41 (9H, s), 1.16-1.10 (1H, m), 0.91 (3H, d), 0.88 (3H, t); 13C NMR (CDCl3, 100 MHz) δ 172.3, 171.1, 155.7, 79.4, 59.1, 56.2, 52.3, 52.1, 47.1, 37.9, 31.1, 29.1, 28.3, 24.6, 24.1, 22.3, 15.6, 11.3, 10.9.
Compound 35
White solid; 1H NMR (CDCl3, 400 MHz) δ 7.44 (1H, s), 7.00 (2H, d), 6.72 (2H, d), 5.43 (1H, d), 4.59 (1H, dd), 4.29 (1H, dd), 3.68 (3H, s), 3.55-3.49 (1H, m), 2.93 (1H, dd), 2.84 (1H, dd), 2.73-2.69 (1H, m), 2.13-2.09 (1H, m), 1.94-1.78 (3H, m), 1.36 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.3, 170.7, 155.66, 130.4, 127.8, 115.4, 80.0, 60.5, 59.4, 58.8, 53.7, 52.6, 46.9, 39.2, 38.6, 31.1, 28.3, 24.4, 21.2.
Compound 36
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.20 (1H, d), 4.48 (1H, dd), 4.38 (1H, dd), 3.85-3.80 (1H, m), 3.67 (3H, s), 3.54-3.43 (2H, m), 2.17-2.02 (2H, m), 1.98-1.91 (2H, m), 1.68-1.65 (1H. m), 1.41 (1H, t), 1.38 (9H, s), 0.94 (3H, d), 0.88 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.6, 171.9, 155.8, 79.5, 59.1, 52.3, 50.4, 46.7, 42.0, 29.1, 28.2, 24.6, 24.2, 23.3, 21.9.
Compound 37
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.22 (1H, d), 4.78 (1H, dd), 4.41 (1H, dd), 3.80-3.76 (1H, m), 3.67 (3H, s), 3.66-3.54 (1H, m), 2.73-2.67 (1H, dd), 2.50 (1H, dd), 2.19-2.14 (1H, m), 2.05-1.90 (3H, m), 1.35 (18H, s); 13C NMR (CDCl3, 100 MHz) δ 172.3, 169.7, 169.5, 154.9, 80.9, 79.8, 59.1, 59.0, 52.6, 52.0, 49.3, 48.9, 46.9, 46.6, 31.0, 29.0, 28.2, 27.9, 24.8, 22.4.
Compound 38
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.39 (1H, s), 4.50 (1H, dd), 3.94 (1H, dd), 3.69 (3H, s), 3.59-3.55 (1H, m), 3.46-3.40 (1H, m), 2.21-2.14 (1H, m), 2.10-1.94 (4H, m), 1.40 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.8, 167.7, 156.2, 80.0, 58.8, 52.7, 46.2, 43.4, 29.4, 28.7, 25.0.
Compound 39
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.66 (1H, d), 4.55 (1H, dd), 4.46 (1H, dd), 4.38 (1H, m), 3.81-3.77 (1H, m), 3.71 (3H, s), 3.67-3.49 (2H, m), 3.37 (1H, s), 2.26-2.20 (1H, m), 2.11-1.99 (1H, m), 2.01-1.91 (2H, m), 1.40 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.6, 169.7, 155.8, 80.1, 63.8, 59.4, 53.5, 52.8, 52.3, 47.1, 30.8, 29.0, 24.7, 22.4.
Compound 40
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.33 (1H, d), 4.57 (1H, dd), 4.36 (1H, dd), 3.79-3.76 (1H, m), 3.64 (3H, s), 3.55-3.48 (1H, m), 2.50-2.42 (2H, m), 2.17-2.12 (1H, m), 2.01 (3H, s), 1.99-1.83 (4H, m), 1.80-1.70 (1H, m), 1.35 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.3, 170.4, 155.3, 79.7, 59.1, 53.4, 51.0, 46.9, 32.6, 31.1, 30.1, 30.8, 24.6, 22.3, 15.5.
Compound 41
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.52 (1H, d), 4.80 (1H, dd), 4.50 (1H, dd), 3.72 (3H, s), 3.67-3.62 (1H, m), 2.78 (2H, t), 2.28-2.20 (1H, m), 2.10-1.92 (4H, m), 1.81-1.77 (1H, m), 1.35 (9H, s), 0.86-0.81 (1H, m); 13C NMR (CDCl3, 100 MHz) δ 172.3, 172.1, 167.3, 154.7, 80.8, 59.3, 53.0, 52.4, 48.8, 47.2, 46.8, 38.6, 29.0, 28.2, 24.7.
Compound 42
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 7.23-7.15 (5H, m), 5.38 (1H, d), 4.61 (1H, dd), 4.25 (1H, dd), 3.66 (3H, s), 3.49-3.45 (1H, m), 3.02 (1H, dd), 2.90 (1H, dd), 2.62-2.57 (1H, m), 1.92-1.75 (3H, m), 1.49-1.43 (1H, m), 1.37 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.3, 170.2, 154.9, 136.4, 129.4, 128.3, 126.8, 79.6, 58.6, 53.5, 52.5, 46.7, 40.3, 28.9, 28.3, 24.3, 22.4.
Compound 43
White solid; 1H NMR (CDCl3, 400 MHz) δ 8.45 (1H, s), 7.58 (1H, d), 7.32 (1H, d), 7.16 (1H, t), 7.09 (1H, t), 7.05 (1H, d), 5.52 (1H, d), 4.70-4.64 (1H, m), 4.15 (1H, dd), 3.67 (3H, s), 3.40-3.34 (1H, m), 3.27 (1H, dd), 3.15 (1H, dd), 2.47-2.41 (1H, m), 1.67-1.44 (3H, m), 1.44 (9H, s), 1.10-1.06 (1H, m); 13C NMR (CDCl3, 100 MHz) δ 172.5, 170.9, 155.1, 136.0, 127.5, 123.1, 121.8, 119.4, 118.6, 111.1, 110.7, 79.6, 58.9, 53.4, 52.3, 46.6, 31.4, 30.0, 29.7, 28.7, 28.2, 24.1.
Compound 44
White solid; 1H NMR (CDCl3, 400 MHz) δ 5.44 (1H, d), 4.48 (1H, q), 4.45 (1H, dd), 3.79-3.74 (1H, m), 3.69 (3H, s), 3.51-3.44 (1H, m), 2.20-2.16 (1H, m), 2.14-1.92 (3H, m), 1.38 (9H, s), 1.30 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 175.5, 172.9, 155.4, 79.5, 49.5, 46.2, 36.9, 35.8, 29.4, 28.3, 18.5.
Compound 45
White solid; 1H NMR (CDCl3, 400 MHz) δ 6.62 (1H, s), 5.82 (1H, s), 5.69 (1H, d), 4.45 (1H, dd), 4.40 (1H, dd), 3.69 (3H, s), 3.59-3.55 (1H, m), 2.31-2.16 (5H, m), 2.05-1.93 (3H, m), 1.79-1.75 (1H, m), 1.40 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 175.4, 172.5, 170.4, 156.1, 80.0, 59.4, 52.3, 51.2, 46.8, 31.5, 29.5, 29.0, 28.2, 24.6, 22.2.
Compound 46
White solid; 1H NMR (CDCl3, 400 MHz) δ 7.49 (1H, s), 7.42 (1H, s), 6.78 (1H, s), 5.63 (1H, d), 4.68 (1H, dd), 4.31 (1H, dd), 3.71 (3H, s), 3.63-3.49 (2H, m), 3.20-3.14 (1H, m), 2.95-2.86 (1H, m), 2.05-2.01 (1H, m), 1.98-1.70 (3H, m), 1.34 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.3, 170.8, 155.2, 135.1, 131.1, 119.6, 79.8, 59.3, 53.4, 52.3, 46.9, 30.3, 28.9, 28.2, 24.6, 22.3.
Compound 47
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.50 (1H, d), 5.10 (1H, dd), 4.43 (1H, dd), 4.35 (1H, dd), 4.15 (1H, q), 3.83-3.78 (1H, m), 3.69 (3H, s), 3.66-3.64 (1H, m), 2.20-2.17 (1H, m), 2.04-1.92 (3H, m), 1.43 (9H, s), 1.15 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.5, 171.0, 156.1, 80.0, 67.5, 59.0, 55.1, 54.0, 52.7, 52.3, 47.2, 38.6, 30.7, 29.0, 24.8, 18.7.
Compound 48
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.33 (1H, d), 4.88 (1H, s), 4.53 (1H, dd), 4.43 (1H, dd), 3.70 (3H, s), 3.61-3.59 (1H, m), 3.11-3.08 (2H, m), 2.21-2.14 (2H, m), 2.07-1.91 (3H, m), 1.76-1.72 (1H, m), 1.61-1.38 (5H, m), 1.40 (18H, s); 13C NMR (CDCl3, 100 MHz) δ 172.4, 171.1, 156.0, 155.5, 79.5, 78.8, 58.6, 52.2, 51.5, 46.8, 40.1, 32.2, 29.3, 28.9, 28.4, 28.3, 24.9, 21.9.
Compound 49
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.10 (1H, d), 4.22 (1H, dd), 4.12 (1H, dd), 3.72-3.764 (1H, m), 3.51 (3H, s), 3.41-3.33 (1H, m), 2.03-1.72 (5H, m), 1.24 (9H, s), 0.77 (3H, d), 0.72 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.2, 170.7, 155.5, 79.0, 58.7, 56.7, 51.9, 46.9, 31.1, 28.9, 28.1, 24.5, 19.5, 19.4, 17.2.
Compound 50
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.23 (1H, d), 4.50-4.48 (2H, m), 3.72-3.65 (2H, m), 3.68 (3H, s), 2.38-2.14 (3H, m), 2.09-1.88 (4H, m), 1.76-1.67 (1H, m), 1.42 (18H, s); 13C NMR (CDCl3, 100 MHz) δ 172.2, 172.1, 170.8, 155.5, 80.4, 79.5, 59.7, 52.1, 50.9, 46.8, 30.7, 28.9, 28.3, 28.0, 27.7, 24.9.
Compound 51
White solid; 1H NMR (CDCl3, 400 MHz) δ 5.44 (1H, d), 4.48 (1H, q), 4.45 (1H, dd), 3.79-3.74 (1H, m), 3.69 (3H, s), 3.50-3.44 (1H, m), 2.21-2.16 (1H, m), 2.12-1.92 (3H, m), 1.38 (9H, s), 1.29 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.6, 171.8, 155.2, 79.6, 59.0, 52.5, 47.9, 46.8, 29.0, 28.3, 24.6, 22.4, 18.7.
Compound 52
White solid; 1H NMR (CDCl3, 400 MHz) δ 8.68 (1H, s), 7.57 (1H, d), 7.32 (1H, d), 7.15 (1H, t), 7.08 (1H, t), 7.02 (1H, d), 5.54 (1H, d), 4.68-4.65 (1H, m), 4.14 (1H, dd), 3.65 (3H, s), 3.38-3.33 (1H, m), 3.26-3.21 (1H, m), 3.15 (1H, dd), 2.46-2.41 (1H, m), 1.65-1.51 (3H, m), 1.44 (9H, s), 1.10-1.06 (1H, m); 13C NMR (CDCl3, 100 MHz) δ 172.5, 170.9, 155.1, 136.0, 127.5, 123.2, 121.8, 111.2, 79.6, 58.9, 53.4, 52.1, 46.6, 38.6, 30.0, 28.7, 24.1.
Compound 53 Light yellow solid; 1H NMR (CDCl3, 400 MHz) δ 7.60 (1H, s), 6.99 (2H, d), 6.72 (2H, d), 5.43 (1H, d), 4.57 (1H, dd), 4.28 (1H, dd), 3.66 (3H, s), 3.52-3.50 (1H, m), 2.93 (1H, dd), 2.83 (1H, dd), 2.72-2.66 (1H, m), 1.96-1.76 (3H, m), 1.56-1.48 (1H, m), 1.40 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.6, 170.7, 155.6, 130.4, 127.7, 115.4, 80.0, 58.9, 53.4, 52.6, 52.3, 46.9, 39.2, 28.9, 28.2, 24.4, 22.3.
Compound 54
White solid; 1H NMR (CDCl3, 400 MHz) δ 6.65 (1H, s), 5.63 (1H, dd), 5.55 (1H, s), 4.49 (1H, dd), 4.41 (1H, dd), 3.71 (3H, s), 3.68-3.55 (2H, m), 2.35-2.14 (3H, m), 2.10-1.95 (4H, m), 1.79-1.71 (1H, m), 1.43 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 175.0, 172.4, 170.2, 156.2, 79.9, 59.1, 52.2, 51.1, 46.7, 31.6, 30.0, 29.1, 28.3, 24.6, 22.2.
Compound 55
White solid; 1H NMR (CDCl3, 400 MHz) δ 4.41 (1H, dd), 4.35 (1H, dd), 3.64 (3H, s), 3.59-3.29 (4H, m), 2.17-1.74 (8H, m), 1.39 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 173.1, 172.4, 154.4, 79.8, 59.1, 58.0, 52.0, 46.5, 46.4, 31.5, 29.8, 28.9, 28.4, 24.9, 23.2, 22.5.
Compound 56
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.20 (1H, d), 4.38 (1H, dd), 4.29 (1H, dd), 3.87-3.81 (1H, m), 3.67 (3H, s), 3.54-3.48 (1H, m), 2.17-1.87 (5H, m), 1.39 (9H, s), 0.92 (3H, d), 0.87 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.4, 170.9, 155.7, 79.3, 58.8, 56.8, 52.3, 47.0, 31.3, 29.1, 28.3, 24.6, 19.6, 17.3.
Compound 57
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.20 (1H, d), 4.47 (1H, dd), 4.37 (1H, dd), 3.84-3.81 (1H, m), 3.65 (3H, s), 3.53-3.44 (2H, m), 2.23-2.02 (2H, m), 1.98-1.91 (2H, m), 1.68-1.62 (1H. m), 1.41 (1H, t), 1.37 (9H, s), 0.94 (3H, d), 0.87 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.3, 171.6, 155.4, 79.4, 58.9, 52.3, 52.1, 50.3, 46.7, 42.6, 29.0, 28.2, 24.6, 24.5, 23.4, 22.3, 21.9.
Compound 58
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.15 (1H, d), 4.40 (1H, dd), 4.31 (1H, dd), 3.87-3.86 (1H, m), 3.67 (3H, s), 3.55-3.49 (1H, m), 2.20-1.92 (4H, m), 1.69-1.64 (1H, m), 1.55-1.49 (1H, m), 1.39 (9H, s), 1.12-1.04 (1H, m), 0.89 (3H, d), 0.86 (3H, t); 13C NMR (CDCl3, 100 MHz) δ 172.3, 171.1, 155.7, 79.4, 58.8, 56.2, 52.2, 52.1, 47.1, 37.8, 31.1, 29.1, 28.3, 24.6, 24.0, 15.6, 11.2.
Compound 59
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 7.22-7.13 (5H, m), 5.38 (1H, d), 4.62 (1H, dd), 4.25 (1H, dd), 3.67 (3H, s), 3.52-3.46 (1H, m), 3.03 (1H, dd), 2.91 (1H, dd), 2.63-2.59 (1H, m), 1.92-1.75 (3H, m), 1.49-1.42 (1H, m), 1.39 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.3, 170.2, 154.9, 136.4, 129.4, 128.3, 126.5, 79.6, 59.2, 53.6, 52.5, 46.7, 40.3, 28.9, 28.3, 28.2, 24.4.
Compound 60
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.65 (1H, d), 4.56 (1H, dd), 4.46 (1H, dd), 3.79-3.52 (4H, m), 3.71 (3H, s), 3.35 (1H, s), 2.27-2.17 (1H, m), 2.09-1.93 (3H, m), 1.40 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.3, 169.7, 155.8, 80.2, 63.9, 59.4, 53.5, 52.8, 52.3, 47.1, 29.0, 28.2, 24.7, 22.4.
Compound 61
White solid; 1H NMR (CDCl3, 400 MHz) δ 7.40 (6H, d), 7.28 (6H, t), 7.21 (3H, t), 5.22 (1H, d), 4.44 (1H, dd), 4.38 (1H, dd), 3.67 (3H, s), 3.61-3.56 (1H, m), 3.13-3.07 (1H, m), 2.59 (1H, dd), 2.42-2.34 (1H, m), 2.13-1.81 (4H, m), 1.42 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.2, 169.1, 154.9, 144.5, 129.6, 127.9, 126.7, 79.7, 66.8, 59.0, 52.2, 51.3, 46.8, 34.5, 29.0, 28.3, 24.6.
Compound 62
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.47 (1H, d), 4.45 (1H, dd), 4.33 (1H, dd), 4.09 (1H, dd), 3.81-3.50 (3H, m), 3.69 (3H, s), 2.25-2.17 (1H, m), 2.07-1.90 (3H, m), 1.40 (9H, s), 1.13 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.8, 170.9, 156.2, 79.9, 67.4, 59.3, 55.2, 54.0, 52.7, 52.2, 47.2, 38.5, 30.6, 29.6, 24.8, 18.4.
Compound 63
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.29 (1H, d), 4.59 (1H, dd), 4.48 (1H, dd), 3.72-3.62 (2H, m), 3.65 (3H, s), 2.54 (2H, t), 2.19-2.14 (1H, m), 2.06 (3H, s), 2.02-1.88 (4H, m), 1.85-1.77 (1H, m), 1.36 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.2, 170.7, 155.4, 79.6, 58.7, 53.4, 50.8, 46.9, 38.5, 32.4, 31.1, 29.7, 28.9, 24.8, 22.1, 15.5.
Compound 64
White solid; 1H NMR (CDCl3, 400 MHz) δ 5.56 (1H, d), 4.59-4.54 (2H, m), 3.87-3.84 (1H, m), 3.82-3.76 (1H, m), 3.72 (3H, s), 3.70-3.63 (2H, m), 3.35 (1H, s), 2.25-2.17 (1H, m), 2.03-1.92 (3H, m), 1.40 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.9, 170.2, 155.6, 80.0, 63.9, 58.9, 53.3, 52.6, 47.2, 31.0, 28.8, 28.3, 24.8, 22.1.
Compound 65
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 7.28-7.17 (5H, m), 5.28 (1H, d), 4.64 (1H, dd), 4.47 (1H, dd), 3.70 (3H, s), 3.61-3.51 (2H, m), 3.18-3.12 (1H, m), 3.08 (1H, dd), 2.90 (1H, dd), 2.15-2.07 (1H, m), 1.94-1.83 (3H, m), 1.37 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.5, 170.8, 155.3, 136.5, 129.9, 128.5, 126.9, 79.8, 60.6, 59.1, 53.4, 52.4, 47.0, 39.4, 29.2, 28.5, 25.0.
Compound 66
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 4.56 (1H, dd), 4.47 (1H, dd), 3.67 (3H, s), 3.61-3.32 (4H, m), 2.20-1.72 (8H, m), 1.41 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.5, 170.8, 156.0, 79.8, 67.4, 58.8, 55.8, 52.4, 47.2, 38.5, 28.8, 28.2, 24.8, 18.6.
Compound 67
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.54 (1H, d), 4.48 (1H, dd), 4.35 (1H, dd), 4.11-4.04 (1H, m), 3.69 (3H, s), 3.74-3.59 (2H, m), 3.57-3.49 (1H, m), 2.21-2.13 (1H, m), 2.02-1.88 (3H, m), 1.37 (9H, s), 1.17 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.5, 170.8, 156.0, 79.8, 67.4, 58.8, 55.8, 52.4, 47.2, 38.5, 28.8, 28.2, 24.8, 18.6.
Compound 68
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.30 (1H, d), 4.54 (1H, dd), 4.45 (1H, dd), 3.72-3.60 (2H, m), 3.63 (3H, s), 2.51 (2H, t), 2.18-2.12 (1H, m), 2.03 (3H, s), 1.99-1.85 (4H, m), 1.82-1.73 (1H, m), 1.33 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.2, 170.7, 155.4, 79.6, 58.6, 52.1, 50.8, 46.9, 38.5, 32.4, 29.7, 28.9, 28.2, 24.8, 24.8, 15.5.
Compound 69
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.13 (1H, d), 4.51 (1H, dd), 4.28 (1H, dd), 3.82-3.76 (1H, m), 3.69 (3H, s), 3.66-3.60 (1H, m), 2.22-2.16 (1H, m), 2.03-1.91 (3H, m), 1.75-1.70 (1H, m), 1.61-1.52 (1H, m), 1.39 (9H, s), 1.14-1.06 (1H, m), 0.99 (3H, d), 0.89 (3H, t); 13C NMR (CDCl3, 100 MHz) δ 172.4, 171.4, 155.7, 79.4, 58.8, 56.2, 52.1, 47.1, 37.9, 29.0, 28.3, 24.9, 24.1, 15.2, 11.2.
Compound 70
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.33 (1H, d), 4.51 (1H, dd), 4.45 (1H, t), 3.69 (3H, s), 3.67-3.63 (1H, m), 3.60-3.54 (1H, m), 2.21-2.14 (1H, m), 2.07-1.88 (3H, m), 1.39 (9H, s), 1.33 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.4, 171.7, 155.2, 79.5, 58.6, 52.2, 47.7, 46.7, 28.9, 28.3, 24.9, 18.2.
Compound 71
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 7.60 (1H, s), 7.06 (2H, d), 6.71 (2H, d), 5.31 (1H, d), 4.61 (1H, dd), 4.49 (1H, dd), 3.69 (3H, s), 3.62-3.58 (1H, m), 3.29-3.24 (1H, m), 3.00 (1H, dd), 2.82 (1H, dd), 2.16-2.10 (1H, m), 1.94-1.87 (3H, m), 1.36 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.3, 171.0, 155.8, 130.7, 127.0, 115.4, 79.9, 59.1, 53.3, 52.2, 46.9, 38.6, 37.9, 28.9, 28.3, 24.8, 22.2.
Compound 72
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 6.57 (1H, s), 5.95 (1H, s), 5.62 (1H, d), 4.49-4.42 (2H, m), 3.69 (1H, m), 3.68 (3H, s), 3.03 (1H, s), 2.31-2.26 (2H, m), 2.22-2.14 (1H, m), 2.11-1.78 (5H, m), 1.37 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 175.7, 172.8, 170.8, 155.9, 79.9, 58.8, 54.1, 52.4, 51.0, 47.0, 31.1,2 8.9, 28.8, 28.2, 24.8.
Compound 73
White solid; 1H NMR (CDCl3, 400 MHz) δ 8.64 (1H, s), 7.67 (1H, d), 7.33 (1H, d), 7.15 (1H, d), 7.11 (2H, t), 5.35 (1H, d), 4.78 (1H, dd), 4.50 (1H, dd), 3.67 (3H, s), 3.55-3.49 (1H, m), 3.27-3.09 (3H, m), 2.14-2.06 (1H, m), 1.91-1.77 (3H, m), 1.38 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.6, 171.2, 155.4, 136.1, 127.8, 123.8, 121.7, 119.3, 118.5, 111.2, 109.7, 79.6, 58.8, 58.7, 52.4, 52.1, 46.8, 38.6, 29.0, 28.5, 28.3, 24.8.
Compound 74
White solid; 1H NMR (CDCl3, 400 MHz) δ 7.43 (6H, d), 7.28 (6H, t), 7.21 (3H, t), 5.07 (1H, d), 4.43 (1H, dd), 4.32 (1H, dd), 3.61 (3H, s), 3.48-3.42 (1H, m), 3.12-3.06 (1H, m), 2.51 (1H, d), 2.15-2.05 (1H, m), 1.95-1.75 (3H, m), 1.56-1.48 (1H, m), 1.40 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.0, 169.5, 155.2, 144.5, 129.7, 127.9, 126.7, 79.8, 66.9, 59.1, 52.1, 51.6, 46.6, 34.1, 28.9, 28.3, 24.7.
Compound 75
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.13 (1H, d), 4.49 (1H, dd), 4.44 (1H, t), 3.76-3.70 (1H, m), 3.66 (3H, s), 3.58-3.53 (1H, m), 2.21-2.08 (1H, m), 2.05-1.89 (3H, m), 1.75-1.69 (1H, m), 1.46 (2H, t), 1.37 (9H, s), 0.95 (3H, d), 0.91 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.4, 171.8, 155.7, 79.4, 58.6, 52.1, 50.3, 46.7, 41.8, 28.9, 28.3, 24.8, 24.5, 23.3, 21.7.
Compound 76
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.19 (1H, d), 4.48 (1H, dd), 4.24 (1H, dd), 3.76-3.70 (1H, m), 3.66 (3H, s), 3.63-3.57 (1H, m), 2.21-2.13 (1H, m), 2.02-1.88 (4H, m), 1.37 (9H, s), 0.98 (3H, d), 0.89 (3H, d); 13C NMR (CDCl3, 100 MHz) δ 172.4, 171.1, 155.8, 79.3, 58.7, 56.8, 52.0, 47.0, 31.2, 28.9, 28.3, 24.9, 19.2, 17.3.
Compound 77
White solid; 1H NMR (400 MHz, CD3OD) δ 7.20-7.42 (15H, m), 4.35 (1H, dd, J=8.4, 4.0 Hz), 4.10 (1H, dd, J=9.2, 5.2 Hz), 3.60 (3H, s), 3.31 (1H, m), 3.00 (1H, m), 2.48 (1H, m), 2.56 (1H, m), 2.15 (1H, m), 1.90 (1H, m), 1.88 (2H, m), 1.44 (9H, br s).
Compound 79
Viscous liquid; 1H-NMR (CD3OD, 400 MHz) δ 4.45 (1H, dd, J=5.6 Hz and 3.2 Hz), 4.30 (1H, m), 4.13 (m, 1H), 3.73 (3H, s), 1.70 (1H, m), 1.53-1.58 (2H, m), 1.48 (9H, brs), 1.17 (3H, d, J=6.4 Hz), 0.96 (3H, d, J=6.4 Hz), 0.94 (3H, d, J=6.4 Hz); 13C NMR (CD3OD, 100 MHz) δ 174.7, 170.8, 156.4, 79.2, 67.1, 57.6, 53.3, 51.4, 40.5, 27.3, 24.5, 22.0, 20.5, 18.8.
Compound 80
White solid; 1H NMR (CD3OD, 400 MHz) δ 4.38 (1H, q, J=6.8 Hz), 4.30 (1H, m), 4.14 (1H, q, J=7.6 Hz), 3.92 (1H, d, J=6.8 Hz), 1.75 (1H, m), 1.53 (1H, m), 1.43 (9H, brs), 1.38 (3H, d, J=6.8 Hz), 1.25 (3H, t, J=7.2 Hz), 1.15 (1H, m), 0.95 (3H, d, J=6.8 Hz), 0.90 (3H, t, J=7.2 Hz); 13C NMR (CD3OD, 100 MHz) δ 172.8, 172.5, 156.4, 79.1, 60.8, 58.8, 48.2, 37.1, 27.3, 24.3, 16.0, 14.4, 13.0, 10.0.
Compound 81
White crystalline solid; 1H NMR (CD3OD, 400 MHz) δ 4.17 (1H, t, J=4.4 Hz), 3.95 (2H, dd, J=17.2, 4.4 Hz), 3.75 (2H, d, J=5.2 Hz), 3.71 (3H, s), 1.45 (9H, brs); 13C NMR (CD3OD, 100 MHz) δ 172.3, 170.3, 156.3, 79.4, 61.9, 56.5, 51.3, 40.5, 27.3.
Compound 82
Viscous liquid; 1H NMR (CD3OD, 400 MHz) δ 4.41 (1H, q, J=6.8 Hz), 4.16 (2H, q, J=7.6 Hz), 3.77 (2H, dd, J=17.6, 4.8 Hz), 1.44 (9H, brs), 1.37 (3H, d, J=7.2 Hz), 1.25 (3H, t, J=6.8 Hz); 13C NMR (CD3OD, 100 MHz) δ 172.7, 170.8, 156.9, 79.2, 60.9, 48.2, 42.9, 27.3, 16.2, 13.0.
Compound 83
White crystal; 1H-NMR (CD3OD, 400 MHz) δ 7.18-7.28 (5H, m), 4.64 (1H, t, J=6.4 Hz), 4.12 (2H, q, J=6.8 Hz), 4.04 (1H, m), 3.15 (1H, dd, J=14.4, 8.0 Hz), 3.0 (1H, dd, J=13.6, 7.6 Hz), 1.62 (1H, m), 1.42 (9H, brs), 1.38-1.36 (2H, m), 1.88 (3H, t, J=7.6 Hz), 0.92 (3H, t, J=6.4 Hz), 0.90 (3H, t, J=6.4 Hz); 13C NMR (CD3OD, 100 MHz) δ 174.0, 171.3.8, 156.2, 136.5, 128.9, 128.0, 126.4, 79.1, 60.9, 53.6, 53.0, 40.8, 37.0, 27.3, 24.4, 21.9, 20.6, 13.0.
Compound 84
White solid; 1H NMR (CD3OD, 400 MHz) δ 7.61 (1H, d, J=7.6 Hz), 7.35 (1H, d, J=7.6 Hz), 7.33 (1H, d), 7.16 (1H, s), 7.11 (2H, t, J=7.2 Hz), 4.50 (1H, t, J=7.6 Hz), 4.41 (1H, dd, J=8.0, 3.2 Hz), 3.68 (3H, s), 3.15-3.20 (1H, m), 2.99-3.10 (3H, m), 2.19-2.16 (1H, m), 1.90-1.83 (3H, m), 1.37 (9H, s); 13C NMR (CD3OD, 100 MHz) δ 172.6, 172.1, 156.2, 136.5, 127.4, 123.7, 121.2, 120.9, 118.5, 110.9, 108.9, 79.1, 59.1, 58.8, 52.8, 51.7, 45.9, 29.6, 28.5, 27.2, 24.3.
Compound 85
Viscous liquid; 1H NMR (CD3OD, 400 MHz) δ 4.46 (1H, dd, J=8.4, 5.2 Hz), 4.20 (1H, d, J=8.4 Hz), 3.90 (1H, m), 3.67 (1H, m), 3.69 (3H, s), 3.66-3.60 (1H, m), 2.27-2.22 (1H, m), 2.06-1.92 (3H, m), 1.78-1.73 (1H, m), 1.63-1.59 (1H, m), 1.42 (9H, brs), 1.12-1.1.07 (1H, m), 0.99 (3H, d, J=7.2 Hz), 0.89 (3H, t, J=6.8 Hz); 13C NMR (CDCl3, 100 MHz) δ 172.5, 172.1, 156.5, 79.0, 59.0, 56.3, 51.1, 47.1, 36.7, 28.7, 27.3, 24.5, 24.2, 14.0, 9.8.
Compound 86
Viscous liquid; 1H NMR (CD3OD, 400 MHz) δ 4.46 (1H, dd, J=4.8, 8.8 Hz), 4.37 (1H, q, J=6.8 Hz), 3.77 (1H, m), 3.69 (3H, s), 3.65-3.62 (1H, m), 3.60-3.54 (1H, m), 2.30-2.23 (1H, m), 2.07-1.93 (3H, m), 1.42 (9H, s), 1.28 (3H, d, J=7.6 Hz); 13C NMR (CDCl3, 100 MHz) δ 172.7, 172.6, 156.2, 79.0, 58.9, 51.3, 47.7, 46.6, 28.5, 27.3, 24.5, 15.7.
Compound 87
Viscous liquid; 1H NMR (CD3OD, 400 MHz) δ 4.36 (1H, d, J=6.0 Hz), 3.73 (2H, brs), 3.71 (3H, s), 2.18-2.10 (1H, m), 1.44 (9H, s), 0.94 (3H, d, J=6.8 Hz), 0.93 (3H, d, J=6.4 Hz); 13C NMR (CD3OD, 100 MHz) δ 173.5, 172.6, 159.2, 80.8, 59.0, 52.6, 44.5, 32.1, 28.8, 19.5, 18.4.
Compound 88
Viscous liquid; 1H NMR (CD3OD, 400 MHz) δ 4.36 (1H, dd, J=5.6, 2.8 Hz), 3.75 (2H, brs), 3.72 (3H, s), 2.20-2.12 (1H, m), 0.96 (3H, d, J=4.4 Hz), 0.94 (3H, d, J=5.2 Hz); 13C NMR (CD3OD, 100 MHz) δ 171.8, 166.1, 57.8, 51.1, 39.9, 30.4, 17.9 16.8.
Compound 90
White solid, mp 86-88 □; [α]D −13.6 (c 1.0, CH3OH); 1H NMR (CD3OD) δ 7.27 (2H, m), 7.20 (1H, m), 7.18 (2H, m), 4.66 (1H, dd, J=8.0, 5.6 Hz), 4.45 (1H, dd, J=7.6, 6.0 Hz), 3.67 (3H, s), 3.66 (3H, s), 3.13 (1H, dd, J=14.0, 5.6 Hz), 3.02 (1H, dd, J=14.0, 8.0 Hz), 2.73 (1H, dd, J=16.4, 6.0 Hz), 2.59 (1H, dd, J=16.4, 7.6 Hz), 1.42 (9H, s); 13C NMR (CD3OD) δ 171.8, 171.6, 171.1, 156.1, 136.4, 128.9, 128.1, 126.5, 79.5, 53.7, 51.3, 51.0, 48.2, 36.9, 35.8, 27.2.
Compound 91
Viscous light yellow liquid; [α]D −3.73 (c 0.54, CH3OH); 1H NMR (CD3OD) δ 8.24 (1H, s), 7.29 (2H, m), 7.23 (1H, m), 7.20 (2H, m), 4.71 (1H, dd, J=8.4, 5.6 Hz), 4.13 (1H, dd, J=8.8, 4.0 Hz), 3.74 (3H, s), 3.70 (3H, s), 3.20 (1H, dd, J=14.0, 5.6 Hz), 3.02 (1H, dd, J=14.0, 8.8 Hz), 2.96 (1H, dd, J=14.0, 4.0 Hz), 2.82 (1H, dd, J=17.6, 8.8 Hz) ppm; 13C NMR (CD3OD) δ 173.1, 171.8, 169.9, 166.8, 138.0, 130.3, 129.8, 128.2, 55.7, 53.1, 53.0, 51.0, 38.2, 36.6.
Compound 92
White solid; mp 88-91 □; [α]D −11.8 (c 1.0, CH3OH); 1H NMR (CD3OD) δ 7.27 (2H, m), 7.20 (1H, m), 7.18 (2H, m), 5.92 (1H, ddt, J=17.2, 10.8, 6.0 Hz), 5.30 (1H, dq, J=17.2, 1.2 Hz), 5.20 (1H, dd, J=10.8, 1.2 Hz), 4.66 (1H, dd, J=7.6, 5.2 Hz), 4.57 (2H, brd, J=6.0 Hz), 4.47 (1H, dd, J=8.0, 5.2 Hz), 3.67 (3H, s), 3.13 (1H, dd, J=13.2, 5.2 HZ), 3.03 (1H, dd, J=13.2, 7.6 Hz), 2.77 (1H, dd, J=16.4, 5.2 Hz), 2.61 (1H, dd, J=16.4, 8.0 Hz), 1.42 (9H, s) ppm; 13C NMR (CD3OD) δ 171.8, 171.6, 170.3, 156.1, 136.4, 132.1, 128.9, 128.1, 126.5, 117.0, 79.5, 65.1, 53.7, 51.3, 51.0, 36.9, 35.8, 27.3.
Compound 93
Viscous liquid; [α]D −4.063 (c 0.58, CH3OH); 1H NMR (CD3OD) δ 8.24 (1H, s), 7.29 (2H, m), 7.22 (1H, m), 7.20 (2H, m), 5.94 (1H, ddt, J=17.2, 10.4, 6.0 Hz), 5.34 (1H, dq, J=17.2, 1.2 Hz), 5.25 (1H, dd, J=10.4, 1.2 Hz), 4.71 (1H, dd, J=8.4, 5.6 Hz), 4.65 (2H, brd, J=6.0 Hz), 4.14 (1H, dd, J=8.8, 4.0 Hz), 3.70 (3H, s), 3.20 (1H, dd, J=14.0, 5.6 Hz), 3.02 (1H, dd, J=14.0, 8.8 Hz), 2.99 (1H, dd, J=14.0, 4.0 Hz), 2.85 (1H, dd, J=18.0, 9.2 Hz); 13C NMR (CD3OD) δ 173.1, 171.1, 169.8, 166.8, 138.0, 133.3, 130.3, 129.8, 128.2, 119.2, 67.3, 55.7, 53.0, 50.9, 38.2, 36.7.
Compound 94
White solid; mp 55-60 □; [α]D −9.16 (c 1.0, CH3OH); 1H NMR (CD3OD) δ 7.34 (2H, m), 7.27 (2H, m), 7.25 (2H, m), 7.21 (1H, m), 7.20 (1H, m), 7.18 (2H, m), 5.11 (2H, s), 4.66 (1H, dd, J=8.0, 5.6 Hz), 4.45 (1H, dd, 7.6, 6.0 Hz), 3.67 (3H, s), 3.13 (1H, dd, 14.0, 5.6 Hz), 3.02 (1H, dd, 14.0, 8.0 Hz), 2.73 (1H, dd, 16.4, 6.0 Hz), 2.59 (1H, dd, 16.4, 7.6 Hz), 1.42 (9H, s); 13C NMR (CD3OD) δ 171.8, 171.6, 170.4, 156.1, 136.4, 135.9, 128.9, 128.1, 128.1, 127.8, 126.5, 126.5, 79.5, 66.2, 53.7, 51.3, 51.0, 36.9, 35.8, 27.2.
Compound 95
Viscous liquid; 1H NMR (CD3OD) δ 8.18 (s), 7.36 (2H, m), 7.27 (4H, m), 7.25 (3H, m), 7.20 (1H, m), 7.20 (2H, m), 5.19 (2H, d, J=2.8 Hz), 4.70 (1H, dd, J=8.8, 5.2 HZ), 4.19 (1H, dd, J=8.8, 4.0 Hz), 3.67 (s), 3.19 (1H, dd, J=14.4, 5.6 Hz), 2.98 (1H, dd, J=14.0, 8.8 Hz), 2.96 (1H, dd, J=18.0, 4.0 Hz), 2.90 (1H, dd, J=18.0, 8.8 Hz); 13C NMR (CD3OD) δ 173.1, 171.6, 169.8, 166.1, 138.0, 137.2, 130.3, 129.8, 129.8, 129.6, 129.6, 128.2, 68.5, 55.7, 53.0, 50.8, 38.1, 36.5
Compound 96
White solid; 1H NMR (CD3OD) δ 7.32 (2H, m), 7.32 (1H, m), 7.32 (2H, m), 4.73 (1H, dd, J=7.6, 5.2 Hz), 4.53 (1H, dd, J=8.0, 6.4 Hz), 3.67 (3H, s), 3.65 (3H, s), 3.11 (1H, dd, J=14.0, 5.2 Hz), 3.01 (1H, dd, J=14.0, 7.6 Hz), 2.74 (1H, dd, J=16.4, 6.4 Hz), 2.55 (1H, dd, J=16.4, 8.0 Hz), 1.89 (3H, s); 13C-NMR (CD3OD) δ 171.6, 171.2, 171.2, 168.6, 136.4, 128.8, 128.1, 127.2, 126.4, 53.8, 51.3, 51.0, 50.1, 36.8, 34.9, 22.5
Compound 97
White solid; 1H NMR(CD3OD) δ 7.79 (2H, dd, J=6.8, 1.6 Hz), 7.57 (1H, tt, J=7.6, 1.2 Hz), 7.47 (2H, t, J=6.8, 8.0), 7.12 (2H, m), 7.12 (1H, m), 7.12 (2H, m), 4.99 (1H, dd, J=7.6, 5.2 Hz), 4.45 (1H, dd, J=8.0, 6.4 Hz), 3.67 (3H, s), 3.66 (3H, s), 3.14 (1H, dd, J=14.0, 5.2 Hz), 3.00 (1H, dd, J=14.0, 7.6 Hz), 2.94 (1H, dd, J=16.4, 6.4 Hz), 2.78 (1H, dd, J=16.4, 8.0 Hz); 13C NMR (CD3OD) δ 171.6, 171.2, 171.2, 168.6, 136.4, 133.4, 131.6, 128.8, 128.1, 128.1, 127.2, 126.4, 53.8, 51.3, 51.0, 50.1, 36.8, 34.9.
Compound 98
White solid; 1H NMR (CD3OD) δ 7.32 (5H, m), 5.92 (1H, ddt, J=17.2, 10.8, 6.0 Hz), 5.30 (1H, dq, J=17.2, 1.2 Hz), 5.20 (1H, dd, J=10.8, 1.2 Hz), 4.80 (1H, dd, J=7.6, 5.2 Hz), 4.66 (1H, dd, J=8.0, 6.4 Hz), 4.57 (2H, brd, J=6.0 Hz), 3.67 (3H, s), 3.14 (1H, dd, J=14.0, 5.2 Hz), 3.00 (1H, dd, J=14.0, 7.6 Hz), 2.83 (1H, dd, J=14.0, 5.2 Hz), 2.64 (1H, dd, J=16.4, 6.4 Hz), 1.89 (3H, s); 13C NMR (CD3OD) δ 171.6, 171.2, 170.3, 168.6, 136.5, 132.1, 129.0, 128.2, 126.6, 117.1, 65.1, 53.8, 51.5, 49.5, 36.9, 35.4, 21.2
Compound 99
Viscous liquid; 1H NMR (CD3OD) δ 7.79 (2H, dd, J=6.8, 1.6 Hz), 7.57 (1H, tt, J=7.6, 1.2), 7.47 (2H, dd, J=6.8, 8.0), 7.11 (5H, m), 5.92 (1H, ddt, J=17.2, 10.8, 6.0 Hz), 5.30 (1H, dq, J=17.2, 1.2 Hz), 5.20 (1H, dd, J=10.8, 1.2 Hz), 4.99 (1H, dd, J=7.6, 5.2 Hz), 4.68 (1H, dd, J=8.0, 6.4 Hz), 4.58 (2H, brd, J=6.0 Hz), 3.70 (3H, s), 3.14 (1H, dd, J=14.0, 5.2 Hz), 3.01 (1H, dd, J=14.0, 7.6 Hz), 2.98 (1H, dd, J=14.0, 5.2 Hz), 2.81 (1H, dd, J=16.4, 6.4 Hz); 13C-NMR (CD3OD) δ 171.6, 171.2, 170.4, 168.6, 136.4, 134.0, 133.4, 131.6, 128.8, 128.1, 128.1, 127.2, 126.4, 119.0, 65.1, 53.8, 51.3, 51.0, 36.8, 35.0
Compound 100
White solid; 1H NMR (CD3OD) δ 7.32 (5H, m), 7.21 (5H, m), 5.11 (2H, s), 4.78 (1H, dd, J=7.6, 5.2 Hz), 4.63 (1H, dd, J=8.0, 6.4 Hz), 3.67 (3H, s), 3.13 (1H, dd, J=14.0, 5.2 Hz), 3.00 (1H, dd, J=14.0, 7.6 Hz), 2.84 (1H, dd, J=14.0, 5.2 Hz), 2.65 (1H, dd, J=16.4, 6.4 Hz), 1.89 (3H, s); 13C NMR (CD3OD) δ 171.8, 171.6, 171.2, 170.2, 168.6, 136.5, 135.9, 128.9, 128.1, 128.1, 127.9, 127.8, 126.5, 66.1, 53.8, 51.3, 49.6, 36.8, 35.4, 21.0
Compound 101
White solid; 1H NMR(CD3OD) δ 7.79 (2H, dd, J=6.8, 1.6 Hz), 7.57 (1H, tt, J=7.6, 1.2 Hz), 7.47 (2H, dd, J=6.8, 8.0 Hz), 7.34 (2H, m), 7.25 (4H, m), 7.20 (1H, m), 7.12 (5H, m), 5.13 (2H, s), 4.99 (1H, dd, J=7.6, 5.2 Hz), 4.45 (dd, J=8.0, 6.4 Hz), 3.69 (s), 3.14 (dd, J=14.0, 5.2 Hz), 3.00 (dd, J=14.0, 5.2 Hz), 2.94 (dd, J=16.4, 6.4 Hz), 2.78 (dd, J=6.4, 8.0 Hz); 13C NMR (CD3OD) δ 171.6, 171.2, 170.5, 168.6, 136.3, 135.9, 133.4, 131.6, 128.8, 128.1, 128.1, 128.1, 127.9, 127.8, 127.2, 126.4, 66.3, 53.8, 51.3, 50.1, 36.8, 35.2
Compound 102
White solid; 1H NMR (CD3OD) δ 7.15 (3H, m), 7.04 (2H, m), 4.65 (1H, dd, J=7.6, 5.2 Hz), 3.79 (1H, dd, J=8.0, 6.4 Hz), 3.61 (3H, s), 3.11 (1H, dd, J=14.0, 5.2 Hz), 3.01 (1H, dd, J=14.0, 7.6 Hz), 2.74 (1H, dd, J=16.4, 6.4 Hz), 2.55 (1H, dd, J=16.4, 8.0 Hz), 1.89 (3H, s); 13C-NMR (CD3OD) δ 173.1, 171.6, 171.1, 170.7, 135.8, 129.1, 128.4, 126.9, 53.5, 52.2, 49.1, 37.4, 35.3, 22.4.
Compound 103
White solid; Melting point 180-190 □ (dec); 1H NMR (CDCl3, 400 MHz) δ 7.23 (1H, brt), 6.65 (1H, d, J=2.2 Hz), 4.39 (1H, t, J=8.8 Hz), 4.08 (1H, dd, J=18, 6.0 Hz) 3.91 (1H, dd, J=18, 6.0 Hz), 3.72 (3H, s), 2.06 (1H, m), 2.0 (3H, s), 0.96 (3H, d, J=7.6 Hz), 0.94 (3H, d, J=7.6 Hz) ppm; 13C NMR (100 MHz, CDCl3) δ 172.2, 170.6, 170.2, 58.5, 52.5, 41.3, 31.3, 23.2, 19.3, 18.5 ppm.
Compound 104
White solid; 1H NMR (CDCl3, 400 MHz) δ 7.17 (1H, brt), 6.55 (1H, d, J=8.4 Hz), 4.56 (1H, m), 4.03 (1H, dd, J=18.4, 6.0 Hz) 3.93 (1H, dd, J=18.0, 5.2 Hz), 3.72 (3H, s), 1.97 (3H, s), 1.66 (2H, m), 1.54 (1H, m), 0.92 (3H, d, J=7.6 Hz), 0.90 (3H, d, J=7.6 Hz) ppm; 13C NMR (100 MHz, CDCl3) δ 172.8, 170.5, 170.1, 52.3, 51.5, 41.1, 41.0, 24.7, 23.0, 22.8, 22.2 ppm.
Compound 105
White solid; 1H NMR (CDCl3, 400 MHz) δ 7.57 (1H, brt), 4.60 (1H, dd, J=7.2 Hz), 3.97 (2H, d, J=6.0 Hz), 3.70 (3H, s), 3.55 (1H, m), 3.41 (1H, m), 2.43 □ 2.38 (1H. m), 2.10 (3H, s), 2.01 □ 1.93 (2H, m), 1.90 □ 1.80 (1H, m) ppm; 13C NMR (100 MHz, CDCl3) δ 171.5, 171.1, 170.1, 59.3, 52.2, 48.2, 41.2, 27.3, 24.9, 22.5 ppm.
Compound 106
Viscous liquid; 1H NMR (CDCl3, 400 MHz) δ 5.39 (1H, s), 4.50 (1H, dd), 3.94 (1H, dd), 3.69 (3H, s), 3.59-3.55 (1H, m), 3.46-3.40 (1H, m), 2.21-2.14 (1H, m), 2.10-1.94 (4H, m), 1.40 (9H, s); 13C NMR (CDCl3, 100 MHz) δ 172.8, 167.7, 156.2, 80.0, 58.8, 52.7, 46.2, 43.4, 29.4, 28.7, 25.0.
Compound 107
Viscous liquid; 1H NMR (CD3OD, 400 MHz) δ 7.52 (1H, d, J=7.6 Hz), 7.33 (1H, d, J=7.6 Hz), 7.10 (1H, d), 7.08 (1H, s), 7.01 (2H, t, J=7.2 Hz), 4.80 (1H, t, J=7.6 Hz), 4.20 (1H, dd, J=8.0, 3.2 Hz), 3.68 (3H, s), 3.39-3.26 (1H, m), 3.22-3.17 (3H, m), 2.38 (H, m), 2.03 (3H, 1H); 13C NMR (CD3OD, 100 MHz) δ 173.6, 169.9, 138.2, 128.7, 124.7, 122.7, 120.0, 119.7, 112.6, 110.6, 61.0, 60.9, 55.4, 53.0, 47.6, 31.2, 28.5, 25.0.
Compound 108
Viscous liquid; 1H NMR (CD3OD, 400 MHz) δ 4.47 (1H, q), 3.70 (3H, s), 3.68 (1H, d), 2.21 (1H, m), 1.42 (3H, s), 1.09 (6H, d); 13C NMR (CD3OD, 100 MHz) δ 174.2, 169,5, 59.7, 52.9, 49.7, 31.7, 18.8, 18.0, 17.3.
Compound 109
White solid; 1H NMR (CD3OD, 400 MHz) δ 4.90 (1H, q, J=6.8 Hz), 4.84 (1H, m), 4.47 (1H, q, J=7.6 Hz), 3.70 (1H, d, J=6.8 Hz), 1.94 (1H, m), 1.61 (1H, m), 1.42 (3H, d, J=6.8 Hz), 1.22 (1H, m), 1.07 (3H, d, J=6.8 Hz), 0.99 (3H, t, J=7.2 Hz); 13C NMR (CD3OD, 100 MHz) δ 174.7, 169.5, 59.1, 52.9, 49.7, 38.3, 25.4, 17.3, 15.1, 11.8.
In order to testify the disease resistance expression in the case of treating the dipeptide derivative of the present invention to the seeding of pepper, PR-1, beta-1,3-glucanase, chitinase, PR4, peroxidase, and PR10 primer, and the seedling of pepper were used. The seedling of pepper was raised in a greenhouse for 4 weeks, Compounds were sprayed thereto in various concentrations, and then, after three or one week, phytophthora (Phytophthora capcisi) and the pathogen of a bacterial rot (Pectobacterium carotovorum) were respectively inoculated.
Since then, after 12, 24, and 48 hours, the respective pepper leaves were collected in an amount of 1 g, and then, stored at an ultralow temperature-room at 75° C. The stored plant leaves were pulverized along with liquid nitrogen, and then, rRNA was extracted by using a RNA extraction kit (Easy-Spin™IIP Total RNA Extraction Kit, Intron Biotechnology, Korea) and Ex Taq polymerase by Kishimoto et al. (2005) (Takara Biomedicals, Otsu, Japan). For amplification, 0.1 μg of cDNA, 10 pMol of each of forward and reverse primers, 250 nM of dNTPs, and 0.5 U of Ex Taq polymerase were added to 20 μl, of a buffer solution. PCR was performed at 94C and 58C in a thermal cycler (PTC-100, USA), and the products by PCR were analyzed under the condition of 1% agarose gel (in 0.5×TAE buffer) (80 V, 60 min). All the RT-PCR tests were repeated twice.
The results thus obtained are illustrated in
After the tobacco (Xanth-nc) bound with GUS gene induced by PR-1α disease-resistance promoter was grown for 3 weeks, 100 μL of the diluted solution of Compound 74 was injected to second leaves with a syringe. After three days, the surrounding leaves were collected in an internal diameter size of 5 mm with a cork borer, the samples thus obtained were added to a 1.5 mL of an eppendorf tube, and 20 μL of GUS extraction buffer solution was further added, ground, and then centrifuged at 8000 G for 3 minutes to obtain supernatant. The same amount of 2 mM MUG (4-methyl umbellifryl-B-glucuronide) solution was added, reacted at 37° C. for 1 hour, and then 960 μL of stop buffer solution (0.2 M, Na2CO3 solution) was added to be 1 mL. Since then, the fluorescence was measured with a TKO 100 fluorometer (Hoefoer Scientific Instruments, USA). At this time, the fluorescence amount was calibrated using a MU (4-methyl umbellyferon) as a standard reagent, and the GUS activity was calculated using MU-mM/sample−10 mg/time. The results thus obtained are listed in the following Table 2 and illustrated in
From the above Table 2, it can be confirmed that as a result of coloring GUS expressed in the case of treating Compound 74 to a tobacco, in the case of treating it in the concentration of 100 ppm, the expression amount is the biggest.
After treating the compounds of the present invention to various plants, the effects of the compounds on suppressing the outbreak of plant disease were measured in the following method.
A tobacco or cucumber was sown in a plastic pot with a diameter of 10 cm×13 cm, and then, 12 pots were added in a yellow box. After seeding, 100 μL was inoculated to the cotyledons of cucumber or tobacco, in which first leaf was started to come out. The test compounds were dissolved in 20% methanol to make a stock solution, and then, the stock solution was diluted to the concentrations of 1, 10, and 100 ppm.
After 7 days, the pathogen of a plant bacterial rot (Pectobacterium carotovorum SCC1) was cultured in a TSA medium for 27 hours; and then, the cultured pathogen was sprayed in a concentration of 108 cfu/mL to the plant leaves, and cultured at 30° C. for 3 days. The plant anthracnose germ (Colletotrichum orbiculare) was cultured in a GBA (Green bean agar) medium to induce spores for about 2 to 3 weeks. The germ was sprayed in a concentration of 105 cell/mL and then cultured at 26° C. for 1 day. When fourth or fifth leaf came out, 200 μL of each of the test compounds was inoculated to third leaf. After 7 days, the pathogen of the plant bacterial rot was sprayed to the plant leaves in the concentration of 108 cfu/mL, and then cultured at 30° C. for 3 days.
The investigation was performed with naked eyes, and the lesion relative areas were investigated until 0 to 100% according to the shortness degrees of the bacterial rot. The anthracnose was investigated by counting the number of lesions occurred on the leaves.
The test results of the effect on suppressing the outbreak of a bacterial rot to a tobacco plant are illustrated in the photographs of
Compound 61 or Compound 74 were deposited on the seeds of cucumbers in the concentration of 100 ppm for 2 hours, and after 3 weeks, the spores of anthracnose germ (Colletotrichum orbiculare) were sprayed in the concentration of 105 spore/mL thereto, and then, after 7 days, the disease outbreak was investigated. The results thus obtained are illustrated in the photographs of
According to
The peppers were raised in the same method as Example 3, and then, the test compounds were dissolved in 20% methanol to make a stock solution, and a stock was diluted to the concentrations of 0.1, 1.0, and 10.0 ppm. The test solutions were sprayed on the leaves or drench-treated on the leaves. After 7 days, the pathogen of plant bacterial rot (Pectobacterium carotovorum SCC1) was cultured in a TSA medium for 27 hours, and then, wetted on an 8 mm diameter paper disc in the concentration of 108 cfu/mL. The paper disc was located in the center of the leaves that were cut roundly, and then, after about 7 days, the lesion area was measured. In the case of phytophthora, the potato agar medium cultured with the phytophthora (Phytophthora capsici) was cut in the internal diameter size of 5 mm with a cork borer, and then inoculated to the leaves. After 7 days, the lesion area was observed. In the case of a bacterial rot, when Compounds 6, 89, 93, and 95 were sprayed on the pepper leaves, the bacterial rot was not found in the concentration of 1 or 10 ppm as compared with the non-treated group. In addition, for the pepper phytophthora test, when Compounds 89, 94, 95, and 107 were drench-treated in the concentration of 1.0 or 10.0 ppm and Compound 107 (1 or 0.1 ppm) was sprayed on the leaves, there were no the lesion formations. Especially, it could be confirmed that Compound 89 exhibited the excellent effect on suppression the outbreak of the pepper phytophthora blight as compared with the control drug, BTH (see
The peppers were raised in the same method as Example 3, Compound 85 was sprayed on the leaves, and then after 1 week, the Phytophthora capsici swarm suspension was drench-inoculated. While observing for 7 days, the lesion formation on the leaves and roots were investigated. As compared with the control group, for the peppers treated with Compound 85, the phytophthora blight on the leaves and roots was significantly suppressed. The test results of Compound 85 on suppressing the outbreak of phytophthora blight are illustrated in
The active ingredients (Compounds 5 to 8, 14, and 15) were drenched to the leaves of a plant (pepper, potato, tomato, tobacco, cucumber, and Chinese cabbage), and then, after 7 days, the plant lengths were measured. Since then, the plant disease pathogen was inoculated, and then, after 3 to 5 days, the sizes of the plant leaves were measured and compared with the control group to obtain the relative growth degrees. The effects on promoting the growth of the cucumber leaves are listed in the following Table 8.
According to the results listed in the above-described Table 8, it can be confirmed that when the dipeptide derivative of the present invention is treated, the growth of the cucumber is promoted. Therefore, it can be confirmed that the compounds of the present invention induce the plant disease-resistance and also the plant growth-promoting effect. It can be also considered that the growth-promoting ability can attenuate plant-protecting ability from the disease.
In order to confirm the immunity-activating effect of the compounds of the present invention, the plants were exposed at the temperature so that they could be artificially damaged from cold weather, and the growth and development were observed. After growing the plants for 6 weeks, the compounds were drenched or sprayed on the leaves and the plants were raised for 1 week. After the plants were exposed under a 2° C. growth room for 1 day, they were raised at room temperature for 3 days. The degrees of the growth and development of the plants were observed and the degrees of the damage from cold weather in the plant were determined. When Compound 95 was drench-treated to the plants, there were no cold weather-damages. When sprayed on the leaves, as with Compounds 90, 91, 93, and 95, there were no damages observed from cold weather in the plant. From the test results, it can be confirmed that when the compounds are applied, the plant exhibits the abiotic resistance and also induces the immunity-activating effect capable of maintaining the growth and development even under the physical harmful environment, like cold weather. The test results are illustrated in the photographs of
Formulation Examples are the representative examples for formulating the agricultural plant-protecting agent including the compound represented by the above-described chemical formula 1 as an active gradient to be suitable for being applied. The compositions of the respective used components used for the formulation are as follows.
10 g of the compound of chemical formula 1, 10 g of NK250L (surfactant), 10 g of white carbon, and 70 g of pyrophylite (bulking agent) were ground and mixed to prepare water dispersible powder.
10 g of the compound of chemical formula 1, 10 g of DDY2000 (surfactant), and 80 g of xylene were mixed to prepare an emulsion.
10 g of the compound of chemical formula 1, 10 g of HY1910 (surfactant), 5 g of propyleneglycol, 0.2 g of xanthan gum, 0.15 g of KM-73 (antifoamer), 0.2 g of Biocide-LS (preservative), 0.1 g of KNP (viscosity agent), and 74.35 g of water (bulking agent) were ground in a ball mill and mixed to prepare the liquid water dispersible powder.
5 g of the compound of chemical formula 1, 7.5 g of paraffin oil, 2 g of sodiumalkylsulfosuccinate (surfactant), 3 g of white carbon, 1.2 g of xanthan gum, 0.8 g of sodium polyacrylic acid, and 80.5 g of potassium chloride were mixed, granulated into a horizontal extruder, and then, dried to prepare the water floating granule.
5 g of the compound of chemical formula 1, 2.5 g of HY1910 (surfactant), 0.2 g of NK250L (surfactant), 0.5 g of soda ash, 2.0 g of dextrin, 25 g of bentonite, and 64.8 g of talc were mixed with water to make the dough, granulated into a horizontal extruder, and then, dried to prepare the granule.
Within the limit range of 20 wt % of the compound of chemical formula 1 used in Formulation Examples 1 to 5 as an active ingredient, the single or mixture selected from the general germicides, insecticides, and herbicides was used to prepare the mixed formulation.
As described above, in the case of treating the agricultural plant-protecting agent of the present invention to various dicotyledoneaes, such as, peppers, cucumbers, potatoes, and tomatoes, the growth of the plant is promoted, and also, the effect on preventing plant diseases such as bacterial rot, damping-off, phytophthora blight, a wilt disease, a leaf spot disease or a mosaic disease, which are caused by a bacteria, a virus, or a mold, is exhibited. Even though it is not directly applied to the lesion area of the plant, the same effect as described above is exhibited on other part of the plant. In addition, since the agricultural plant-protecting agent of the present invention exhibits the effect on preventing the cold weather-damage so as for the plant to be not damaged from a low temperature, it exhibits the plant immunity-activating effect so as to make the plant stays to grow healthy.
Therefore, the agricultural plant-protecting agent of the present invention can be applied for a plant (in detail, dicotyledoneae) for preventing or suppressing the plant disease, promoting the plant growth, or activating the plant immunity.
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2012-0040279 | Apr 2012 | KR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/KR2013/003303 | 4/18/2013 | WO | 00 |
