STRUCTURE
A three-dimensional stereochemistry formula (1) of 6R-(3,6-dideoxy-L-arabinohexopyranosyloxy)heptanoic acid as a pheromone compound isolated from C. elegance is determined according to spectroscopic analysis such as HR-MASS, IR, DEPT, 2D-NMR (HMBC, HMQC, NOE, ROESY, and TOCSY).
A pure molecular weight of the pheromone, 6R-(3,6-dideoxy-L-arabino-hexopyranosyloxy)heptanoic acid, is 276 dalton, and a monocular formula of the pheromone is C13H24O6. A calculated high-resolution mass number of the pheromone is 276.1651. It is noted that a high-resolution mass number measured by a high resolution-FAB is 276.1652, and this mass number is almost identical to the calculated mass number (see FIG. 1 ). Functional groups of relative carbonyl and hydroxy groups of the pheromone molecule are identified by an infrared (IR) analysis (see FIG. 2 ).
In order to determine the three-dimensional stereochemistry configuration of the novel pheromone compound of formula (I), 2D-proton.nuclear magnetic resonance spectrum (1H-NMR) is measured by using dutro-methanol (CD3OD) as a solvent. A C-13 nuclear magnetic resonance spectrum (13C-NMR) is also measured by using dutro-methanol (CD3OD) as a solvent. The chemical shift is represented by ppm.
After the location of each carbon is identified by 1H-NMR (see FIG. 3 ), 13C-NMR (see FIG. 4 ) and DEPT (see FIG. 5 ), the chemical shift of 1H— and 13C— is measured by using HMBC (see FIG. 6 ), HMQC (see FIG. 7 ), ROESY (see FIG. 8 ), and TOCSY (see FIG. 9 ) spectrums to identify the accurate relation of the 1H— and 13C. Table 4 shows a result of HMBC spectrum.
In order to measure the stereo interrelation in the three-dimensional space, the two-dimensional NMR technology of NOE is used. FIGS. 10 through 12 show the obtained NOE spectrum.
The 6R-(3,6-dideoxy-L-arabino-hexopyranosyloxy)heptanoic acid of the stereochemistry formula (I) is obtained by a coupling reaction of reactants represented in formulas (II) and (III).
2,4-di-O-benzoyl-3,6-dideoxy-L-arabino-hexopyranose of formula (II) is synthesized as shown in the following reaction formula 1 from L-rhamnose monohydrate of formula (IV).
The compound (XI) (472.9 mg, 0.976 mmol) is dissolved in MeOH (20 ml) NaOMe (52.7 mg, 0.976 mmol) is added at 0° C. The temperature is gradually increased to a room temperature and the mixture is stirred for 12 hours. After the reaction is finished, MeOH is vacuum concentrated. Then, in order to eliminate a sub-product methylbenzoate, it is dissolved in water (20 ml) and washed by CH2Cl2 (20 ml×5). The pH of the solution layer is adjusted using amberlite resin type acid such as amberlite and amberlite IR-120 (H+) (500 mg). After the filtration, the water is removed from the solution layer by freeze drying method, and then the compound (I) (234.6 mg, 87%) is isolated using flash column chromatography (EtOAc/MeOH, 11:1, v/v).
I; a colorless oil, Rf=0.43 (EtOAc/MeOH, 11:1, v/v);
[α]D20=−81.0 (c=0.1, MeOH);
IR(film) Vmax 3391, 2969, 2933, 1712, 1452, 1379, 1244, 1126, 1103, 1042, 1031 cm−1;
1H NMR (500 MHz CD3 OD) δ 4.64 (s, 1H, H-1′), 3.80-3.77 (m, 1H, H-6), 3.72-3.71 (m, 1H, H-2′), 3.63-3.59 (m, 1H, H-5′), 3.54-3.49 (m, 1H, H-4′), 2.30 (t, 2H, J=7.5 Hz; H-2), 1.96-1.92 (m, 1H, H-3′ eq), 1.79-1.74 (m, 1H, H-3′ax), 1.61 (m, 2H, H-3), 1.56-1.50 (m, 2H, H-5), 1.47 (m, 2H, H-4), 1.21 (d, 3H, J=6.5 Hz H-6′), 1.12 (d, 3H, J=6.5 Hz H-7);
13C NMR (125.7 MHz; CD3OD) δ 177.7 (C-1), 97.6 (C-1′, α), 72.4 (C-6), 71.3 (C-5′), 70.1 (C-2′), 68.5 (C-4′), 38.2 (C-5), 36.1 (C-3′), 35.0 (C-2), 26.5 (C-3), 26.1 (C-4), 19.4 (C-7), 18.2 (C-6′);
An HRMS (FAB) calculated value for C13H25O6) (M++H) m/z is 277.1651, and an actual measured value is 277.1652.
| TABLE 4 | |||
| (Spectrum analysis result of pheromone, 6R-(3.6-dideoxy-L- arabino-hexopyranosyloxy) | |||
| heptanoic acid | |||
| Position | δH (mult, J ) | δC | HMBC (H to C) |
| 1 | 177.3 | 2, 3 | |
| 2 | 2.30 (t, 7.5) | 34.6 | 1, 3 |
| 3 | 1.64 (m) | 25.5 | 2, 4, 5, 6 |
| 4 | 1.47 (m) | 25.1 | 2, 3, 5 |
| 5 | 1.50-1.48 (m) | 37.1 | 3, 4, 6, 7 |
| 6 | 3.80-3.77 (m) | 71.3 | 5, 7, 1′ |
| 7 | 1.14 (d, 6.5) | 18.3 | 5, 6 |
| 1′ | 4.66 (s) | 96.6 | 2′, 3′, 6 |
| 2′ | 3.73-3.72 (m) | 69.0 | 1′, 3′ |
| 3′ | 1.97-1.95 (m) | 34.9 | 1′, 4′, 5′ |
| 1.79-1.74 (m) | |||
| 4′ | 3.54-3.59 (m) | 67.4 | 3′, 5′, 6′ |
| 5′ | 3.64-3.62 (m) | 70.2 | 3′, 4′, 6′ |
| 6′ | 1.24 (d, 6.5) | 17.2 | 4′, 5′ |
As described above, the present invention firstly determined stereochemistry configuration of pheromone, (6R)-6-(3.6-dideoxy-L-arabino-hexopyranosyloxy) heptanoic acid and salts thereof. Based on this fact, the effective total synthesis was successfully performed, thereby overcoming the minute isolation of the pheromone obtained from C. elegance to make it possible to mass-produce the pheromone.
Accordingly, it becomes possible to develop medical substances using the pheromone relating to aging, stress, metabolism, signal transfer system in vivo, and anti-cancer, obesity and a suppressing agent for aging and stress. In addition, it becomes also possible to research the active target protein body of the pheromone.
US 7951839 B2
http://www.google.com/patents/US7951839
KDR Biotech Co. Ltd
506-4, Amsa 2-Dong,
Kangdong-Ku, Seoul,
134-052, Korea.
Kangdong-Ku, Seoul,
134-052, Korea.
