6R-(3,6-dideoxy-L-arabinohexopyranosyloxy)heptanoic acid as a pheromone compound isolated from C. elegance

 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).

Embodiment 10 Synthesis of 6R-(3,6-dideoxy-L-arabino-hexopyranosyloxy)heptanoic acid (I)

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′

INDUSTRIAL APPLICABILITY

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.

6R-(3,6-dideoxy-L-arabino-hexopyranosyloxy)heptanoic acid, preparation process for the same and dauer effect thereof
US 7951839 B2

http://www.google.com/patents/US7951839


DESCRIPTION OF DRAWINGS

FIG. 1 is a HR-MS-FMB spectrum of a pheromone of stereochemistry formula (1) according to the present invention;

FIG. 2 is an IR spectrum of the pheromone according to the present invention;

FIG. 3 is a ¹H-NMR spectrum of the pheromone according to the present invention;

FIG. 4 is a ¹³C-NMR spectrum of the pheromone according to the present invention;

FIG. 5 is a ¹³C-NMR DEPT spectrum of the pheromone according to the present invention;

FIG. 6 is a 2D-NMR HMBC spectrum of the pheromone according to the present invention;

FIG. 7 is a 2D-NMR HMQC spectrum of the pheromone according to the present invention;

FIG. 8 is a 2D-NMR ROESY spectrum of the pheromone according to the present invention;

FIG. 9 is a 2D-NMR TOCSY spectrum of the pheromone according to the present invention;

FIG. 10 is a 2D-NMR NOE(1) spectrum of the pheromone according to the present invention;

FIG. 11 is a 2D-NMR NOE(2) spectrum of the pheromone according to the present invention; and

FIG. 12 is a 2D-NMR NOE(3) spectrum of the pheromone according to the present invention.

KDR Biotech Co. Ltd

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Kangdong-Ku, Seoul,
134-052, Korea.

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