DEUTERIUM-LABELED CANNALACTONE, SYNTHESIS AND USE FOR THE DETERMINATION OF CANNALACTONE IN ALL TISSUES OR EXUDATES OF PLANTS OR LIVING ORGANISMS

Abstract

The present invention relates to the chemical synthesis of cannalactone, an adaptation of this synthesis leading to deuterium labeling of cannalactone, and the use of deuterium-labeled cannalactone to perform the assay of natural cannalactone from exudates or tissues of the hemp plant.

Description

Title of the invention: DEUTERIUM-LABELED CANNALACTONE, SYNTHESIS AND USE FOR THE DETERMINATION OF CANNALACTONE IN ALL TISSUES OR EXUDATIONS FROM PLANTS OR LIVING ORGANISMS Technical field of the invention

[0001] The present invention relates to the chemical synthesis of cannalactone, an adaptation of this synthesis leading to deuterium labeling of cannalactone, as well as the use of deuterium-labeled cannalactone to perform the assay of cannalactone in all tissues or exudates of plants or living organisms, and in particular in the exudates or tissues of the hemp plant by liquid chromatography coupled with mass spectrometry. Technical background

[0002] Hemp (Cannabis saliva) is an annual plant native to Asia that has been cultivated for over 8,000 years. It is grown worldwide and is capable of meeting the four basic needs of humanity: food, shelter, clothing, and medicine.

[0003] Hemp cultivation is widespread because it is a profitable and sustainable crop. Indeed, the plant adapts easily to diverse soils and climates and therefore requires no phytosanitary treatment. The area under hemp production in France has increased thirtyfold between 1960 and today. This interest in hemp continues to grow, partly with the aim of gradually replacing cotton, which is very water-intensive. France is currently the leading hemp producer in Europe, with production on approximately 20,000 hectares.

[0004] Known for its resistance to parasites and pests, the hemp plant is all the more attractive. However, a parasitic plant of the broomrape family, branched broomrape, Phelipanche ramosa, induces large yield losses in hemp crops, sometimes exceeding 80%. This parasite also attacks crops such as rapeseed or tobacco, but a specialization of a branched broomrape population to hemp has been demonstrated [1]-[3].

[0005] After germination, the parasitic plant connects to the root of the host plant and thus obtains nutrients for its own development [4]. This parasitism causes significant damage to hemp crops.

[0006] Strigolactones are small molecules known to be exuded into the soil at picomolar concentrations (1012 M) and have been identified as stimulants germination of the seeds of Phelipanche ramosa or branched broomrape [5], [6]. Strigolactones (SL) were first identified for their role in parasitic and symbiotic interactions in the rhizosphere and are the most recent class of plant hormones to have been discovered [7], [8]. They are best known for their role in controlling plant architecture; more recently, roles for SL in other aspects of plant development have been highlighted [9].

[0007] The Applicant has identified that cannalactone, which is a strigolactone exuded by hemp, was responsible for initiating the parasitic life cycle of branched broomrape [1],

[10] ,

[11] .

[0008] To this end, he developed a chemical synthesis allowing access to cannalactone (low bioavailability), as well as an adaptation of this synthesis for obtaining a cannalactone labeled with deuterium and its use for carrying out the assay of natural cannalactone from exudates or tissues of the hemp plant. Summary of the invention

[0009] In particular, the present invention relates to deuterium-labeled cannalactone, characterized in that it conforms to the general formula 1:

[0010] [Chem.l]

[0011] in which:

[0012] - R1 designates a hydrogen atom H or a deuterium atom D,

[0013] - R2 designates the methyl radical CH3 or the tri-substituted deuterated methyl radical CD3,

[0014] - at least one of R1 and R2 being deuterated.

[0015] Advantageously, R1 can designate a hydrogen atom H and R2 can designate the tri-substituted deuterated methyl radical CD3, so that the deuterium-labeled cannalactone according to the invention will correspond to the following formula 2:

[0016] [Chem.2]

[0017] Advantageously, R1 can designate a deuterium atom D and R2 can designate the methyl radical CH3, so that the deuterium-labeled cannalactone according to the invention will correspond to the following formula 3:

[0018] [Chem.3]

[0019] Advantageously, R1 can designate a deuterium atom D and R2 can designate the tri-substituted deuterated methyl radical CD3, so that the deuterium-labeled cannalactone according to the invention will correspond to the following formula 4:

[0020] [Chem.4]

[0021] The present invention also relates to a method for synthesizing cannalactone, of the following general formula 5:

[0022] [Chem.5]

[0023] characterized in that it comprises the following steps:

[0024] - a reaction A) of coupling commercial B-cyclocitral with a C4 bromofuran of the following formula 6:

[0025] [Chem.6] ..c ,.>~onps IM' a

[0026] to obtain a B20 alcohol of the following formula 7:

[0027] [Chem.7] B2G

[0028] - a step B) of reducing alcohol B20 of formula 7, in particular by a hydride, to obtain a mixture of diastereomers of allylic alcohol, followed by a separation step (in particular by silica chromatography) of said diastereomers to retain the (4R* 6R*)-B21 diastereomer of the following formula 8:

[0029] [Chem. 8] ​​r°

[0030] - a step C) of selective epoxidation of the compound (47?*, 67?*)-B21 of formula 8, in particular directed by allylic alcohol, to obtain the (47?*, 67?*)-cz'5-B22 epoxide of the following formula 9:

[0031] [Chem.9] SH'J-cis B22

[0032] - a radical reaction D) of reduction and isomerization of the epoxide (47?*, 6 7?*)-cz'5-B22 of formula 9, to obtain the tertiary alcohol (47?*, 87?*)-Bl of the following formula 10:

[0033] [Chem. 10] —•O

[0034] - a step E) of protection of the tertiary alcohol (47?*, 87?*)- B1 of formula 10, for obtain the protected alcohol (47?*, 87?*)-B23 of the following formula 11:

[0035] [Chem. 11] (4 / r. ««*^823

[0036] - a step F) comprising the formalization Fl) of the compound (47?*, 87?*)-B23 of formula 11 using alkyl formate (in particular ethyl or methyl formate) of the following formula 12:

[0037] [Chem. 12]

[0038] then the O-alkylation F2) of the aldehyde thus formed using compound D4 of the following formula 13:

[0039] [Chem. 13] \ ... / EM

[0040] leading to the obtaining of the mixture of diastereomers (4R*, 67?*)-B24 of the following formula 14:

[0041] [Chem. 14] (W, 8R*}-BZ4

[0042] - a step G) of deprotection of the tertiary alcohol and separation (in particular by silica chromatography) of diastereomers of formula 14, leading to the isolation of cannalactone of formula 5.

[0043] The process for synthesizing cannalactone according to the invention is illustrated below by [Fig.1] which shows the ten-step access route developed by the inventors for chemically synthesizing cannalactone.

[0044] The present invention also relates to a method for synthesizing a deuterium-labeled cannalactone according to the invention described above, characterized in that it comprises the following steps:

[0045] - the steps A to E of synthesis of the protected compound (47?*, 87?*)-B23 of formula 11 such as described previously for the synthesis of non-deuterated cannalactone (steps A to E are strictly identical);

[0046] - a step F) comprising the formalization Fl) of the compound (47?*, 87?*)-B23 of formula 11 using alkyl formate (in particular ethyl or methyl formate) corresponding to formula 12

[0047] or

[0048] using deuterated alkyl formate (in particular deuterated ethyl or methyl formate) corresponding to the following formula 15:

[0049] [Chem. 15]

[0050] - then the O-alkylation of the aldehyde thus formed using compound D4 (in particular that described in publication

[12] ) of the following formula 13:

[0051] [Chem. 13] y EM

[0052] or using the deuterated compound D4 (in particular that described in the publication

[0053]

[0054]

[0055]

[0056]

[0057]

[0058]

[0059]

[0060]

[0061]

[0062]

[0063]

[0064]

[0065]

[0066]

[13] ) of the following formula 16: [Chem. 16] œr; 04 -dfntîfïrè at least one of the organic compound functionalized with a formyl group or of compound D4 being deuterated, to obtain a mixture of deuterated diastereomers (47?*, 6R*)-B24 of formula 17 next: [Chem. 17] with - R1 designating a hydrogen atom H or a deuterium atom D, and - R2 designates a methyl group (CH3) or a methyl group where the hydrogens have been replaced by deuterium atoms (CD3), - at least one of R1 and R2 being deuterated; - a step G) of deprotection of the tertiary alcohol and separation of the diastereomers (in particular by chromatography on silica), leading to the isolation of the deuterium-labeled cannalactone of formula 1. The process for synthesizing deuterium-labeled cannalactone according to the invention is also illustrated below by [Fig. 1] Advantageously, according to a first advantageous embodiment of the process for synthesizing a deuterium-labeled cannalactone, the formylation Fl) can be carried out using the alkyl formate (in particular ethyl formate or methyl formate) corresponding to formula 12, and the O-alkylation F2) of the aldehyde following the formylation Fl) can be carried out using the compound D4 of formula 16. In this case, this embodiment will lead to the formation of a deuterium-labeled cannalactone corresponding to formula 2. [Chem. 2] ( jüH Advantageously, according to a second advantageous embodiment of the process for synthesizing a deuterium-labeled cannalactone, the formylation Fl) can be carried out using alkyl formate (in particular ethyl formate or methyl formate) corresponding to formula 15, and the O-alkylation F2) of the aldehyde following the formylation Fl) is carried out using compound D4 of formula 13. In this case, this embodiment will lead to the formation of a deuterium-labeled cannalactone corresponding to formula 3.

[0067] [Chem.3]

[0068] Advantageously, according to a third advantageous embodiment of the process for synthesizing a deuterium-labeled cannalactone, the formylation Fl) can be carried out using deuterated alkyl formate (in particular deuterated methyl formate or deuterated ethyl formate) corresponding to formula 15, and the 1'0 -alkylation F2) of the aldehyde following the formylation Fl) is carried out using compound D4 of formula 16. In this case, this embodiment will lead to the formation of a deuterium-labeled cannalactone corresponding to formula 4.

[0069] [Chem.4]

[0070] Finally, the present invention further relates to the use of deuterium-labeled cannalactone according to the invention or as obtained according to the process according to the invention relating to obtaining deuterium-labeled cannalactone, for carrying out the determination of cannalactone in all tissues or exudates of plants or living organisms, and in particular in the exudates or tissues of the hemp plant, and in particular for carrying out the determination of natural cannalactone from the exudates or tissues of the hemp plant by liquid chromatography coupled with mass spectrometry. Brief description of the figures

[0071] Other features and advantages of the invention may become apparent to a person skilled in the art upon reading the examples below, given by way of illustration and not limitation and illustrated by the attached [Fig. 1]:

[0072] [Fig.l] - the [Fig.l] represents the scheme of the total racemic synthesis of cannalactone labeled or not with deuterium. EXAMPLES Solvents and reagents

[0073] The chemical reagents are commercial products marketed in particular by the companies Sigma Aldrich, Alfa Aesar, Acros Organics and TCI. They were used without further purification.

[0074] The analytical-grade dry solvents are commercial products marketed by companies including Sigma Aldrich and Acros Organics. Tetrahydrofuran (THF) was distilled under argon over sodium in the presence of benzophenone. The deuterated solvents are marketed by Eurisotop. Materials and Methods

[0075] The non-aqueous reactions were carried out under an inert atmosphere (argon or nitrogen), using standard techniques for handling air- and moisture-sensitive compounds.

[0076] All reactions were monitored by thin-layer chromatography (TLC) on pre-coated silica gel aluminium plates (marketed by Merck under the trade name 60 F254 with short-wavelength UV detection (i.e. X = 254 nm), and / or by staining with a solution of KMnO4 [1% (w / w)] in water or a solution of vanillin [1% (w / w)] in a 1% (v / v) ethanoic acid solution.

[0077] Most of the separations were carried out under Flash-silica gel chromatography conditions using a packed cartridge (40-63 µm silica gel) at medium pressure (20 psi) with an Armen fraction collector or a pump or a Buchi Pure C-805 Flash.

[0078] Some separations were carried out on preparative thin layer chromatography (PTLC) (Merck 60 F254 silica gel on glass).

[0079] The *H NMR spectra were recorded on Bruker spectrometers at 300, 500, or 700 MHz. The 13C NMR spectra were recorded on the same instruments at 75, 125, or 175 MHz. The chemical shifts δ are expressed in parts per million (ppm) with the residual solvent signals as an internal reference (δ = 7.24 for *H NMR and 77.23 for 13C NMR in CDC13). For *H NMR, the spectra are described as follows: chemical shift, integration, multiplicity (s = singlet, d = doublet, t = triplet, q = quadruplet, quint = quintuplet, sext = sextuplet, dd = doublet of doublet, dt = doublet of triplet, m = multiplet), coupling constant in Hertz (J), and assignment. All NMR assignments are based on COSY, HSQC, and HMBC experiments. NOESY experiments were recorded to confirm the double-bond configurations.

[0080] IR spectra were recorded on a PerkinElmer Spectrum 100 FT-IR spectrometer, absorptions being given in centimetres1 (cm1).

[0081] Low-resolution mass spectra were determined by electron fogging ionization on a Waters Acquity UPLC system, combined with a photodiode detector (PDA), an evaporative light scattering detector (ELSD), and a mass spectrometer with a tandem quadrupole detector (TQD). The buffers and aqueous mobile phases for UPLC were prepared using purified water with a Milli-Q system.

[0082] High-resolution mass spectra were obtained with the Waters Acquity UPLC device (by direct injection or with a BEH C[8 2.1 Â ~50 mm, 1.7 pm column) combined with a PDA and a Waters LCT Premier XE mass instrument (ESI with a time-of-flight (ToF) analyzer).

[0083] The measurements of the optical rotation were recorded on an Anton Paar MCP 300 polarimeter using a 1 dm long cell.

[0084] Circular dichroism measurements were recorded on a JASCO J-810 spectropolarimeter at a scan speed of 50 nm / min in acetonitrile from 200 to 400 nm.

[0085] The chiral separation of synthetic cannalactone enantiomers was carried out on a THAR supercritical fluid apparatus. This instrument, marketed under the name "Investigator II", consists of a pump module for CO2 as well as for the co-solvent, a sample changer, a pressure regulator and a collector that can hold up to 6 fractions.

[0086] Detection was performed using an iodine-band UV spectrometer (commercially available under the trade name Waters PDA 2998n, 200 to 800 nm). The instrument parameters were optimized for the separation of the racemic mixture. The column used was a Daicel IC (immobilized silica based on cellulose protected by tris(3,5-dichlorophenylcarbamate)) column, 4.6 x 250 mm in size and with a particle size of 5 µm. The furnace temperature was set at 25 °C and the system pressure was 100 bar. The total flow rate was set at 4 mL / min, with 10% (v / v) methanol. The enantiomers were separated into two flasks using a booster pump at a fixed flow rate of 3 mL / min of methanol. The collection conditions are summarized in Table 1 below:

[0087] [Tables 1] Wavelength (nm) Start Threshold (mAu) Stop Threshold (mAu) Start Time Window (min) Stop Time Window (min) Vial 1 226 20 30 22.8 27.5 Vial 2 226 20 20 22.8 33

[0088] EXAMPLE 1: Synthesis of natural cannalactone according to the first process of the invention (access route illustrated by [Fig. 1]) 4-Bromofuran-2(5H)-one (C8)

[0089] Oxalyl dibromide (2.6 g, 10.00 mmol) in CH2Cl2 (22 mL) and DMF (1 mL) was added to a solution of furan-2,4(3 / 1,5H)-dione (1.0 g, 10.00 mmol) in CH2Cl2 (22 mL) and DMF (1 mL) at 0 °C. The mixture was stirred for 1 h at 0 °C and gradually warmed to room temperature for 2 h. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3 x 20 mL). The combined organic phases were washed with water (2 x 30 mL), a saturated aqueous solution of NaHCO3 (2 x 30 mL), and brine (2 x 30 mL) and dried over Na2SO4. The solvents were removed to obtain the crude product C8 (1.61 g, quantitative) as a brown solid. The chemical analyses are in agreement with the literature

[14] . (4-B romofuran-2-yl)oxy triisopropylsilane (C4)

[0090] To a solution of 4-bromofuran-2(5 / / )-one (C8) (720.4 mg, 4.40 mmol) in CH2C12 (6.2 mL) under argon at 0 °C, Et3N (626.4 mg, 6.20 mmol, 1.4 equiv.) was added. The mixture was stirred for 1 minute, then triisopropylsilyl trifluoromethanesulfonate (TIPSOTf) (1.42 g, 4.60 mmol, 1.05 equiv.) was added dropwise at 0 °C. The resulting mixture was stirred for 10 minutes at 0 °C, then warmed to room temperature and stirred for another 1.5 hours. The mixture was diluted with heptane (10 mL), washed with saturated aqueous NaHCO3 solution (2 x 10 mL), water (2 x 10 mL), and brine (2 x 10 mL). The organic phase was dried over Na2SO4. The solvents were removed to obtain C4 bromofuran (1.4 g, quantitative) as a brown oil. The chemical analyses are consistent with the literature

[14] .

[0091] 4-[Hydroxy(8,12,12-trimethylcyclohex-7-en-6-yl)methyl]furan-2(5H)-one (B20)

[0092] To a solution of C4 (89.9 mg, 0.28 mmol) in dry THF (1.8 mL) under argon at -78 °C, a solution of n-BuLi (0.3 mL, 0.30 mmol, 0.98 M, 1.1 equiv.) is added dropwise. The resulting mixture was stirred at -78 °C for 30 minutes. A mixture of [3-cyclocitral (51.6 mg, 0.34 mmol, 1.2 equiv.) in dry THF (2 mL) is then added. The reaction mixture was stirred for 2 h at -78 °C and 12 h at room temperature. The mixture was hydrolyzed with a saturated aqueous solution of NH₄Cl (5 mL) and an aqueous solution of HCl (5 mL, 2 M). The organic phase was separated, and the aqueous phase was extracted with EtOAc (3 x 5 mL). The combined organic phases were washed with water (2 x 5 mL), a saturated aqueous solution of NaHCO₃ (2 x 5 mL), water (2 x 5 mL), and brine (2 x 5 mL), then dried over Na₂SO₄. The solvents were removed, and the mixture was purified by silica gel chromatography (heptane / EtOAc, 95:5 to 60:40 for 20 min) to obtain the pure product B20 (24.5 mg, 37%) in the form of a brown oil, of formula 7:

[0093] B20

[0094] [Chem.7] s -O 2 14 Chemical Formula; ChHjqOj ,--15 exact Mass: 238.1412 “—Tu Moiacular Weight: 236.31 W 15 & 50

[0095] 'H NMR (500 MHz, CDC13) ô 5.91 (1H, d, J = 1.5 Hz, H-3), 5.10 (1H, s, H-6), 4.88 (1H, d, J = 18.0 Hz, H-5a), 4.71 (1H, d, J = 18.0 Hz, H-5b), 1.96 (2H, t, J = 6.0 Hz, H-9), 1.61 (3H, s, H-15), 1.59-1.55 (2H, m, H-10), 1.50-1.46 (2H, m, H-ll), 1.13 (3H, s, H-13 or H-14), 0.98 (3H, s, H-13 or H-14).

[0096] 13C NMR (75 MHz, CDC13) ô 174.1 (C-2), 174.0 (C-4), 138.7 (C-7), 136.6 (C-8), 115.0 (C-3), 71.9 (C-5), 67.8 (C-6), 39.5 (C-ll), 35.0 (C-12), 33.7 (C-9), 28.9 (C-13 or C-14), 28.5 (C-13 or C-14), 21.4 (C-15), 19.3 (C-10).

[0097] IR (film) vmax 3471, 2932, 1777, 1741, 1637, 1447, 1268, 1111, 1028 cm"1.

[0098] HRESIMS m / z 237.1491 [M + H]+ (wedge, for C14H21O3, 237.1491).

[0099] 4-[Hydroxy(8,12,12-trimethylcyclohex-7-en-6-yl)methyl]dihydrofuran-2(3 H )- one (B21)

[0100] To a solution of B2O (696.4 mg, 2.95 mmol) in methanol (45 mL) at 15 °C, NiCl2 (350.9 mg, 1.48 mmol, 0.5 equiv.) and then sodium borohydride (358.3 mg, 9.47 mmol, 3.2 equiv.) were added in portions. The mixture was stirred at 15 °C until TLC analysis indicated complete conversion. The reaction mixture was hydrolyzed with an aqueous solution of HCl (50 mL, 2 M). The aqueous phase was extracted with CH2Cl2 (3 x 20 mL). The combined organic phases were dried over Na2SO4 and the solvents were removed. The resulting mixture was then purified by silica gel chromatography (CH2Cl2 / EtOAc, 100:0 to 90:10 for 30 min) to obtain the pure product (47?*, 67?*)-B21 of formula 8 (327.4 mg, 47%) as a yellow oil and (47?*, 6S*)-B21 of formula 18 (131.9 mg, 19%) as a white solid:

[0101] (47?*, 67?*)-B21

[0102] [Chem.8] [ ¥=O HO. Chemical Formula: 14 As4 », Exact Mass: 238,1859 43 \AxZ ’ Moiecuîsr Wejght 2% 50

[0103] RMN ‘H (500 MHz, CDC13) ô 4.50 (1H, dd, J = 9.5, 7.0 Hz, H-5a), 4.28 (1H, dd, J = 9.5, 7.0 Hz, H-5b), 4.21 (1H, d, J = 9.5 Hz, H-6), 3.21 (1H, sext, J = 9.5 Hz, H-4), 2.41 (1H, dd, J = 17.5, 8.5 Hz, H-3a), 2.18 (1H, dd, J = 17.5, 8.5 Hz, H-3b), 1.96 (2H, q, J = 5.5 Hz, H-9), 1.81 (3H, s, H-15), 1.59-1.52 (2H, m, H-10), 1.48-1.45 (1H, m, H-lla), 1.40-1.35 (1H, m, H-llb), 1.08 (3H, s, H-13 ou H-14), 0.98 (3H, s, H-13 ou H-14).

[0104] RMN 13C (125 MHz, CDC13) ô 177.1 (C-2), 138.4 (C-7), 135.1 (C-8), 72.9 (C-5), 72.6 (C-6), 41.3 (C-4), 40.4(C-ll), 35.0 (C-12), 34.6 (C-9), 32.4 (C-3), 29.2 (C-13 ou C-14), 29.1 (C-13 ou C-14), 21.3 (C-15), 19.4 (C-10).

[0105] IR (film) vmax 3464, 2928, 1768, 1551, 1365, 1263, 1178, 1048, 1001, 892 cm1.

[0106] HRESIMS m / z 239.1640 [M + H]+ (cale, pour C14H23O3, 239.1647).

[0107] (4R*, 65*)-B21

[0108] [Chem. 18] I x=o HO. Chemisai Formula' « 7]$ 4 3, Mass; 238,1569 ,, '5 Motecular Weitint 238,3270 J 112 '8 “ 1t>

[0109] RMN‘H (500 MHz, CDC13) ô 4.17 (1H, dd, J = 9.0, 7.0 Hz, H-5a), 4.16 (1H, d, J = 9.0 Hz, H-6), 3.94 (1H, dd, J = 9.0, 7.0 Hz, H-5b), 3.19 (1H, sext, J = 9.0 Hz, H-4), 2.72 (1H, dd, J = 17.5, 7.5 Hz, H-3a), 2.57 (1H, dd, J = 17.5, 7.5 Hz, H-3b), 1.96 (2H, q, J = 5.0 Hz, H-9), 1.80 (3H, s, H-15), 1.59-1.51 (2H, m, H-10), 1.47-1.44 (1H, m, H-lla), 1.40-1.34 (1H, m, H-llb), 1.08 (3H, s, H-13 ou H-14), 0.97 (3H, s, H-13 ou H-14).

[0110] RMN 13C (125 MHz, CDC13) ô 177.5 (C-2), 138.2 (C-7), 135.1 (C-8), 72.3 (C-6), 70.5 (C-5), 41.5 (C-4), 40.4 (C-ll), 35.1 (C-12), 34.6 (C-9), 33.8 (C-3), 29.2 (C-13 ou C-14), 28.8 (C-13 ou C-14), 21.4 (C-15), 19.4 (C-10). [YES] IR (film) vmax 3481, 2925, 2870, 1774, 1547, 1465, 1373, 1258, 1176, 1092, 1033, 1011,890, 795 cm'1.

[0112] HRESIMS m / z 239.1638 [M + H] + (calc. for Ci4H23O3 239.1647).

[0113] (4 R *)-[(6 R *)-hydroxy(8,12,12-trimethyl-7-oxabicyclo[4.1.0]heptan-6- yl)methyl]dihydrofuran-2(3H)-one ((4 / ?*,6R*)-cis-B22)

[0114] A solution of (47?*, 6R*)-B21 (100.9 mg, 0.420 mmol) in dry toluene (5.1 mL) was added to a solution of VO(acac)2 (3.9 mg, 0.015 mmol, 0.04 equiv.) in dry toluene (0.2 mL). Tert-Butyl hydroperoxide (TBHP) (0.11 mL, 5.5 M, 0.590 mmol, 1.4 equiv.) was then added. The resulting mixture was stirred at room temperature for 1 h. The reaction mixture was hydrolyzed with an aqueous NaOH solution (5 mL, 5%). The aqueous phase was extracted with heptane and of EtOAc (2:1) (3 x 10 mL). The combined organic phases were washed with brine (2 x 10 mL), dried over Na2SO4, and the solvents were removed to obtain the pure product (47?*, 67?*)-cA-B22 of formula 9 (116.4 mg, quantitative) as a colorless oil used in the next step without purification:

[0115] (47?*, 67?*)-cA-B22

[0116] [Chem.9] HO,, $ Chemical Formula; 54 j®,OS Exact Mass-. 254.1518 n Molecular Weight 254.3260

[0117] RMN‘H (300 MHz, CDC13) ô 4.40 (1H, dd, J = 9.5, 8.0 Hz, H-5a), 4.31 (1H, dd, J = 9.5, 8.0 Hz, H-5b), 3.96 (1H, d, J = 8.0 Hz, H-6), 2.94 (1H, sext, J = 8.0 Hz, H-4), 2.59 (1H, dd, J = 17.0, 9.0 Hz, H-3a), 2.48 (1H, dd, J = 17.0, 9.0 Hz, H-3b), 1.90-1.80 (1H, m, H-9a), 1.78-1.69 (1H, m, H-9b), 1.39 (3H, s, H-15), 1.36-1.32 (2H, m, H-10), 1.25-1.22 (1H, m, H-lla), 1.06 (3H, s, H-13 ou H-14), 1.05-1.03 (1H, m, H-llb), 1.02 (3H,

[0118] s, H-13 ou H-14).

[0119] RMN 13C (75 MHz, CDC13) ô 176.4 (C-2), 71.1 (C-5), 70.5 (C-6), 70.4 (C-7), 66.3 (C-8), 40.0 (C-4), 37.6 (C-ll), 33.9 (C-13), 33.3 (C-3), 31.8 (C-9), 25.6 (C-13 et C-14), 22.2 (C-15), 17.0 (C-10).

[0120] IR (film) vmax 3464, 2928, 1768, 1551, 1365, 1263, 1178, 1048, 1001, 892 cm’1.

[0121] HRESIMS m / z 255.1607 [M + H]+(calc. pour C14H23O4, 255.1596)

[0122] (4 R *)-( Z )-[(8 R *)-hydroxy-(8,12,12-triméthylcyclohexylidene)méthyl ]dihydrofuran-2(3H)-one ((4 R *, 8 R *)-Bl)

[0123] A solution of titanocene dichloride (298.7 mg, 1.20 mmol, 5.0 equiv.) and manganese (197.8 mg, 3.60 mmol, 15.0 equiv.) in strictly deoxygenated dry THF (1.2 mL) under argon was stirred for 1 h in a round-bottom flask with a Rodavis stopper. The solution changed from red to green. To this mixture, a solution of (47?*, 67?*)-cA-B22 (61.1 mg, 0.24 mmol) in strictly deoxygenated dry THF (1.2 mL) under argon was added. The resulting mixture was stirred for 22 h at room temperature. The reaction mixture was hydrolyzed with a saturated aqueous solution of NaH2PO4 (3 mL). The aqueous phase was extracted with EtOAc (3 × 5 mL). The organic phase was washed with a saturated aqueous solution of NaHCO3 (2 x 5 mL) and brine (2 x 5 mL), dried with Na2SO4, and the solvents were removed. The crude product was purified by silica gel chromatography. (heptane / EtOAc, 85:15 to 70:30 for 20 min) to obtain a mixture of (47?*, 6R *)-cA-B22, (47?*, 6R*)-B21 and (47?*, 87?*)-Bl (58.2 mg).

[0124] The mixture is then diluted in pyridine (0.5 ml) and Ac2O is added (10 drops). This reaction was stirred for 48 h at room temperature until TLC analysis indicated complete conversion. The crude product was purified by silica gel chromatography (heptane / EtOAc, 90:10 to 70:30 for 20 min) to obtain the pure product (47*, 87*)-Bl of formula 10 (11.4 mg, 20%) as a colorless oil:

[0125] (47?*, 87?*)-Bl

[0126] [Chem. 10] Chemical Formula: C^HjgOj Jj Éxaet Mass: 238.1569 ta -Aw «4^ Motecuiar Wsight: 238.3270

[0127] RMN ‘H (500 MHz, CDC13) ô 5.25 (1H, d, J = 9.5 Hz, H-6), 4.46 (1H, t, J = 8.0 Hz, H-5a), 4.30 (1H, sext, J = 8.0, H-4), 3.89 (1H, t, J = 8.0 Hz, H-5b), 2.61 (1H, dd, J = 17.5, 9.0 Hz, H-3a), 2.18 (1H, dd, J = 17.5, 9.0 Hz, H-3b), 1.84-1.79 (1H, m, H-9a), 1.59-1.55 (2H, m, H-10), 1.54-1.51 (1H, m, H-9b), 1.42 (3H, s, H-15), 1.39-1.34 (2H, m, H-ll), 1.12 (3H, s, H-13 ou H-14), 1.05 (3H, s, H-13 ou H-14).

[0128] RMN 13C (125 MHz, CDC13) ô 177.7 (C-2), 154.9 (C-7), 124.7 (C-6), 74.5 (C-5), 74.1 (C-8), 44.2 (C-9), 39.7 (C-ll), 37.3 (C-12), 36.4 (C-3), 36.0 (C-4), 32.3 (C-13 ou C-14), 32.2 (C-13 ou C-14), 30.7 (C-15), 19.4 (C-10).

[0129] IR (film) vmax3490, 2931, 2854, 1775, 1463, 1368, 1256, 1172, 1091, 1007, 872, 788 cm

[0130] HRESIMS m / z 239.1639 [M + H]+ (cale, pour C14H23O3, 239.1647).

[0131] (4 R *)-[( Z )-{(8 R *)-(8,12,12-triméthyl-8[(triméthylsilyl)oxy]cyclohe xylidenejmethyl}] dihydrofuran-2(3 H )-one ((4 7? *, 8 R *)-B23)

[0132] A mixture of crude alcohol (47?*, 87?*)-Bl (94.3 mg, 0.24 mmol) and trimethylsilylimidazole (TMS-imidazole) (1.06 mL, 7.20 mmol, 30.0 equiv.) was stirred at 50 °C under argon overnight. The mixture was cooled to room temperature, stirred for 1 h, and diluted with heptane (5 mL). The organic phase was washed with brine (2 x 5 mL), dried with Na₂SO₄, and the solvents were removed. The crude product was purified by silica gel chromatography (heptane / EtOAc, 90:10 to 60:40 for 20 min) to obtain the pure product (47?*, 87?*)-B23 of formula 11 (14.6 mg, 20% in 2 steps) in the form of a yellow oil:

[0134] [Chem. 11] 1 ï g Chemical Formula: CvHsoO$Si 1<^ I] Exact Mass: 31 0.1964 w —Moiecuiar Wetghî: 310.6090:1 s> US

[0135] RMN‘H (500 MHz, CDC13) ô 5.18 (1H, d, J = 9.5 Hz, H-6), 4.41 (1H, t, J = 8.0 Hz, H-5a), 4.31 (1H, sext, J = 9.5 Hz, H-4), 3.88 (1H, t, J = 8.0 Hz, H-5b), 2.58 (1H, dd, J = 17.0, 8.5 Hz, H-3a), 2.15 (1H, dd, J = 17.0, 8.5 Hz, H-3b), 1.86-1.81 (1H, m, H-9a), 1.74 (1H, td, J = 13.0, 4.5 Hz, H-9b), 1.59-1.53 (2H, m, H-10), 1.42 (3H, s, H-15), 1.40-1.32 (2H, m, H-ll), 1.12 (3H, s, H-13 ou H-14), 1.04 (3H, s, H-13 ou H-14), 0.12 (9H, s, H-TMS).

[0136] RMN 13C (125 MHz, CDC13) ô 177.7 (C-2), 155.1 (C-7), 124.1 (C-6), 77.5 (C-8), 74.3 (C-5), 42.6 (C-9), 39.4 (C-ll), 37.3 (C-12), 36.3 (C-3), 35.7 (C-4), 32.8 (C-13 ou C-14), 32.7 (C-13 ou C-14), 32.1 (C-15), 19.3 (C-10), 3.2 (C-TMS).

[0137] IR (film) vmax 2963, 2928, 1781, 1469, 1366, 1250, 1250, 1162, 1066, 1035, 1012, 838 cm1.

[0138] HRESIMS m / z 311.1952 [M + H]+(calc. pour Ci7H31O3Si, 311.2042). (4R*, 8R*)-B24

[0139] To a solution of (47?*, 87?*)-B23 (11.2 mg, 0.04 mmol) in dry THF (0.4 mL) at -40 °C under argon, ethyl formate (32 wt.L, 0.4 mmol, 10.0 equiv.) and tert-BuOK (32.5 mg, 0.28 mmol, 7.0 equiv.) were added. The mixture was stirred for 1 h at 0 °C. The mixture was diluted in EtOAc (2 mL), washed with water (2 x 2 mL) and brine (2 x 2 mL), dried over Na2SO4, and concentrated under reduced pressure to obtain the crude enol (7.5 mg, 55%). This compound was used without further purification in the next step.

[0140] To a crude enol solution (7.5 mg, 0.02 mmol) in dry THF (0.2 mL) at -78 °C, tert-BuOK (3.4 mg, 0.03 mmol, 1.5 equiv.) and a solution of 5-bromo-3-methylfuran-2(5H)-one D4

[12] (5.8 mg, 0.03 mmol, 1.5 equiv.) in dry THF (0.2 mL) were added. The reaction mixture was warmed to room temperature and stirred overnight. The reaction mixture was diluted in EtOAc (3 mL), washed with water (2 x 2 mL) and brine (2 x 2 mL), then dried with Na2SO4, and the solvents were removed. The mixture was purified by PTLC (petroleum ether / EtOAc, 80:20) to obtain the pure product (47?*, 87?*)-B24 of formula 17 (2.1 mg, 12% in 2 steps) in the form of a yellow oil:

[0142] [Chem. 17] Chemicà! Fomwfe: Exact Mass; 434.2125 Motecufar Weigh-: 434.604-0

[0143] Isomer 1:

[0144] RMN ‘H (700 MHz, CDC13) ô 7.43 (1H, d, J = 3.0 Hz, H-6’), 6.79 (1H, t, J = 1.5 Hz, H-3’), 6.06 (1H, s, H-2’),5.22 (1H, dd, J = 13.5, 10.0 Hz, H-6), 4.88-4.84 (1H, m, H-4), 4.47 (1H, dd, J = 9.0, 3.0 Hz, H-5a), 3.91 (1H, sext, J = 5.5 Hz, H-5b), 1.97 (3H, t, J = 1.5 Hz, H-7’), 1.85-1.81 (1H, m, H-lla), 1.73-1.68 (1H, m, H-llb), 1.61-1.54 (2H, m, H-10), 1.46 (3H, s, H-15), 1.39-1.34 (2H, m, H-9), 1.11 (3H, s, H-13 ou H-14), 1.08 (3H, s, H-13 ou H-14), 0.13 (9H, s, H-TMS).

[0145] RMN 13C (175 MHz, CDC13) ô 172.3 (C-2), 170.4 (C-5’), 152.7 (C-7), 150.6 (C-6’), 140.9 (C-3’), 136.0 (C-4’), 124.0 (C-6), 113.3 (C-3), 100.6 (C-2’), 77.7 (C-8), 72.8 (C-5), 43.0 (C-ll), 39.7 (C-9), 37.3 (C-12), 37.2 (C-4), 32.8 (C-13 ou C-14), 32.7 (C-13 ou C-14), 30.9 (C-15), 19.5 (C-10), 10.9 (C-7’), 3.4 (C-TMS).

[0146] Isomère 2 :

[0147] RMN ‘H (700 MHz, CDC13) ô 7.45 (1H, d, J = 2.5 Hz, H-6’), 6.83 (1H, t, J = 1.5 Hz, H-3’), 6.06 (1H, s, H-2’), 5.22 (1H, dd, J = 13.5, 10.0 Hz, H-6), 4.88-4.84 (1H, m, H-4), 4.47 (1H, dd, J = 9.0, 3.0 Hz, H-5a), 3.91 (1H, sext, J = 5.5 Hz, H-5b), 1.95 (3H, t, J = 1.5 Hz, H-7’), 1.85-1.81 (1H, m, H-lla), 1.73-1.68 (1H, m, H-llb), 1.61-1.54 (2H, m, H-10), 1.45 (3H, s, H-15), 1.39-1.34 (2H, m, H-9), 1.02 (3H, s, H-13 ou H-14),0.9 (3H, s, H-13 ou H-14), 0.12 (9H, s, H-TMS).

[0148] RMN 13C (175 MHz, CDC13) ô 172.3 (C-2), 170.4 (C-5’), 152.7 (C-7), 150.9 (C-6’), 141.0 (C-3’), 135.8 (C-4’),123.9 (C-6), 113.2 (C-3), 100.7 (C-2’), 77.6 (C-8), 72.6 (C-5), 43.1 (C-ll), 39.7 (C-9), 37.3 (C-12), 37.1(C-4), 32.5 (C-13 ou C-14), 32.1 (C-13 ou C-14), 30.8 (C-15), 19.5 (C-10), 10.8 (C-7’), 3.4 (C-TMS).

[0149] IR (film) vmax 2928, 2851, 17887, 1734, 1681, 1463, 1376, 1337, 1250, 1184, 1081, 1031, 1006, 956, 838 cm1.

[0150] HRESIMS m / z 249.1482 [M + H - H2O]+ (hold, for C15H21O3, 249.1491). (±)-2'-epi-cannalactone and (±)-cannalactone

[0151] To a solution of (47?*, 8R*)-B24 (30.1 mg, 0.070 mmol) in CH3CN (0.7 mL) and water (5 drops), a solution of Sc(OTf)3 (0.3 mg, 7 pmol, 1 mol%) in CH3CN (0.7 mL) was added. The resulting mixture was stirred for 1.5 h at room temperature and hydrolyzed with phosphate buffer (1.5 mL, pH 7). The organic phase was extracted with CH2C12 (3 x 2 mL), and the combined organic phases were The samples were washed with brine (2 x 2 mL), dried over Na2SO4, and the solvents were removed to obtain the crude product. The crude product was purified by PTLC (petroleum ether / EtOAc, 60:40) to obtain the product (±)-2'-epz-cannalactone (8.3 mg, 33% in 3 steps) as a yellow oil and (±)-cannalactone of formula 5 (6.9 mg, 27% in 3 steps) as a yellow oil. (±)-2'-epicannalactone

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158] i Chemteat Focmufa: Exact Mass: 362,1729 Motecutar Weight. 362,4220 RMN‘H (700 MHz, CDC13) ô 7.43 (1H, d, J = 3.0 Hz, H-6’), 6.81 (1H, t, J = 1.5 Hz, H-3’), 6.08 (1H, t, J = 1.5 Hz, H-2’), 5.32 (1H, d, J = 9.5 Hz, H-6), 4.83 (1H, tt, J = 9.0, 2.5 Hz, H-4), 4.52 (1H, t, J = 9.0 Hz, H-5a), 3.94 (1H, dd, J = 9.0, 5.5 Hz, H-5b), 1.98 (3H, t, J = 1.5 Hz, H-7’), 1.83-1.79 (1H, m, H-9a), 1.59-1.55 (2H, m, H-10), 1.53-1.48 (1H, m, H-9b), 1.43 (3H, s, H-15), 1.39-1.34 (2H, m, H-ll), 1.11 (3H, s, H-13 ou H-14), 1.02 (3H, s, H-13 ou H-14). RMN 13C (175 MHz, CDC13) ô 172.3 (C-2), 170.4 (C-5’), 152.5 (C-7), 150.5 (C-6’), 140.9 (C-3’), 136.1 (C-4’), 124.9 (C-6), 113.4 (C-3), 100.5 (C-2’), 73.9 (C-8), 73.2 (C-5), 44.6 (C-9), 40.0 (C-ll), 37.3 (C-4), 37.2(C-12), 32.2 (C-13 ou C-14), 31.8 (C-13 ou C-14), 29.6 (C-15), 19.6 (C-10), 10.9 (C-7’). IR (film) vmax 3493, 2928, 2848, 1785, 1751, 1684, 1465, 1382, 1347, 1179, 1088, 1031,954 cm’1. HRESIMS m / z 363.1813 [M + H]+ (cale, pour C20H27O6, 363.1808). (±)-cannalactone [Chem. 5] 1 Chemical Formula: Exact Mass: 362,1729 Motecutar Welght 362,4220 RMN ‘H (700 MHz, CDC13) ô 7.47 (1H, d, J = 2.5 Hz, H-6’), 6.85 (1H, t, J = 1.5 Hz, H-3’), 6.07 (1H, t, J = 1.5 Hz, H-2’), 5.31 (1H, d, J = 9.5 Hz, H-6), 4.83 (1H, tt, J = 9.0, 3.0 Hz, H-4), 4.52 (1H, t, J = 9.0 Hz, H-5a), 3.92 (1H, dd, J = 9.0, 6.0 Hz, H-5b), 1.95 (3H, t, J = 1.0 Hz, H-7’), 1.82-1.78 (1H, m, H-9a), 1.57-1.53 (2H, m, H-10), 1.51-1.47 (1H, m, H-9b), 1.43 (3H, s, H-15), 1.38-1.32 (2H, m, H-ll), 1.07 (3H, s, H-13 ou H-14), 0.9 (3H, s, H-13 ou H-14).

[0159] RMN 13C (175 MHz, CDC13) ô 172.2 (C-2), 170.4 (C-5’), 152.6 (C-7), 151.1 (C-6’), 141.0 (C-3’), 135.7 (C-4’), 124.5 (C-6), 113.2 (C-3), 100.7 (C-2’), 73.8 (C-8), 72.9 (C-5), 44.6 (C-9), 39.9 (C-ll), 37.3 (C-4), 37.1(C-12), 32.1 (C-13 ou C-14), 31.3 (C-13 ou C-14), 29.4 (C-15), 19.5 (C-10), 10.7 (C-7’).

[0160] IR (film) vmax 3479, 2919, 2854, 1784, 1747, 1678, 1466, 1384, 1340, 1182, 1082, 1031, 1007,951 cm1.

[0161] HRESIMS m / z 363.1794 [M + H]+ (cale, pour C20H27O6, 363.1808).

[0162] (+)-cannalactone [a]D 24 + 15.00 ± 1.59 (c 0.12, CHC13)

[0163] (-)-cannalactone [a]D24 - 11.73 ± 2.53 (c 0.13, CHC13)

[0164] EXAMPLE 2: synthesis of deuterium-labeled cannalactone according to the second process of the invention (access route illustrated by Figure 2)

[0165] The following products are the same as those prepared in example 1

[0166] - 4-bromofuran-2(577)-one (C8)

[0167] - (4-bromofuran-2-yl)oxytriisopropylsilane (C4)

[0168] - 4-[hydroxy(8,12,12-trimethylcyclohex-7-en-6-yl)methyl]furan-2(577)-one (B20)

[0169] - 4-[hydroxy(8,12,12-trimethylcyclohex-7-en-6-yl)methyl]dihydrofuran-2(377)-one (B21)

[0170] - (47?*)-[(67?*)-hydroxy(8,12,12-trimethyl-7-oxabicyclo[4.1.0]heptan-6-yl) methyl] dihydrofuran-2(377)-one ((47?*, 67?*)-cA-B22)

[0171] - (47?*)-(Z)-[(87?*)-hydroxy-(8,12,12-trimethylcyclohexylidene)methyl ]dihydrofuran-2(377)-one ((4 / ?*, 87?*)-Bl)

[0172] - (47?*)-((Z)-[(87?*)-(8,12,12-trimethyl-8{(trimethylsilyl)oxy]cyclohex ylidene)methyl]) dihydrofuran-2(377)-one ((47?*, 87?*)-B23).

[0173] To a solution of (47?*, 87?*)-B23 of formula 11 (11.0 mg, 0.035 mmol) in dry THF (0.35 mL) at -40 °C under argon, ethyl formate (28 pL, 0.35 mmol, 10.0 equiv.) and tert-BuOK (29.8 mg, 0.25 mmol, 7.0 equiv.) were added. The mixture was stirred for 1 h at 0 °C. The mixture was diluted in EtOAc (2 mL), washed with water (2 x 2 mL) and brine (2 x 2 mL), dried with Na2SO4, and concentrated under reduced pressure to obtain the crude enol (11.4 mg, 97%).

[0174] To a solution of crude enol (11.4 mg, 0.034 mmol) in dry THF (0.34 mL) at -78 °C, tert-BuOK (6.1 mg, 0.05 mmol, 1.5 equiv.) and a solution of deuterated compound D4

[13] (10.2 mg, 0.05 mmol, 1.5 equiv.) in dry THF (0.34 mL) are added. The reaction mixture is allowed to warm to room temperature and stirred overnight. The mixture was diluted in EtOAc (3 mL), washed with water (2 x 2 mL) and brine (2 x 2 mL), dried with Na2SO4, and the solvents were removed. obtain the crude compound (47?*, 8R*)-B24-D3 (21.2 mg) of formula 17 (alternative in which R1 is a hydrogen atom and R2 is the CD3 group):

[0175] (4R* 8R*)-B24-D3

[0176] [Chem. 17]

[0177] HRESIMS m / z 438.2393 [M + H] + (calc. for C23H32D3O6, 438.2391).

[0178] A solution of (4 / ?*, 8R*)-B24-D3 (14.9 mg, 0.034 mmol) in CH3CN (0.34 mL) and water (5 drops) was added to a solution of Sc(OTf)3 (1.9 mg, 4 pmol, 10 mol%) in CH3CN (0.34 mL). The resulting mixture was stirred for 1.5 h at room temperature and hydrolyzed with phosphate buffer (1.5 mL, pH 7). The organic phase was extracted with CH2C12 (3 x 2 mL), and the combined organic phases were washed with brine (2 x 2 mL), dried with Na2SO4, and the solvents were removed to obtain the crude product. The crude product was purified by silica gel chromatography (petroleum ether / EtOAc, 70:30 to 50:50 for 15 min) to obtain the product (±)-2'-epz-cannalactone-D3 (3.9 mg, 31% in 3 steps) as a yellow oil and (±)-cannalactone-D3 of formula 1 (alternative in which R1 is a hydrogen atom and R2 is the CD3 group) (4.1 mg, 32% in 3 steps) as a yellow oil. EXAMPLE 3#: Biological results

[0179] We have carried out the determination of cannalactone in different varieties of hemp using cannalactone according to the invention according to formulas 2, 3 or 4 according to the protocol described in pea

[13] . We were able to quantify it. Bibliographical references

[0180] 1. Hamzaoui, O. et al., Proceedings of the 15th World Congress on Parasitic Seedlings; Amsterdam, The Netherlands 32, (2019). 2. Stojanova, B., Delourme, R., Duffé, P., Delavault, P. & Simier, P. Genetic differentiation and host preference reveal non-exclusive host races in the generalist parasitic weed Phelipanche ramosa. Weed Res. 59, 107-118, doi: 10.1111 / wre. 12353 (2019). 3. Huet, S., Pouvreau, J.-B., Delage, E., Delgrange, S., Marais, C., Bahut, M., Delavault, P., Simier, P. & Poulin, L. Populations of the Parasitic Plant Phelipanche ramosa Influence Their Seed Microbiota. Forehead. Plant Sci. 11, 1075, doi: 10.3389 / fpls.2020.01075 (2020). 4. Delavault, P., Montiel, G., Brun, G., Pouvreau, J. B., Thoiron, S. & Simier, P. Communication Between Host Plants and Parasitic Plants. Adv. Bot. Res. 82, 55-82, doi: 10.1016 / bs.abr.2016.10.006 (2017). 5. Cook, C. E.; Whichard, L. P.; Turner, B.; Wall, M. E.; Egley, G. H. Germination of Witchweed (Striga Lutea Lour.): Isolation and Properties of a Potent Stimulant. Science. 154 (3753), 1189-1190. doi:10.1126 / science. 154.3753.1189 (1966). 6. Daignan Former, S.; Keita, A.; Boyer, F.-D., Chemistry of Strigolactones, Key Players in Plant Communication. ChemBioChem doi:10.1002 / cbic.202400133 (2024). 7. Gomez-Roldan, V., Fermas, S., Brewer, P. B., Puech-Pages, V., Dun, EA, Pillot, J.-P., Letisse, F., Matusova, R., Danoun, S., Portais, J.-C., Bouwmeester, H., Bécard, G., Beveridge, CA, Rameau, C. & Rochange, SF Strigolactone inhibition of shoot branching. Nature 455, 189-194, doi:10.1038 / nature07271 (2008). 8. Umehara, M., Hanada, A., Yoshida, S., Akiyama, K., Arite, T., Takeda-Kamiya, N., Magome, H., Kamiya, Y., Shirasu, K., Yoneyama, K., Kyozuka, J. & Yamaguchi, S. Inhibition of shoot branching by new terpenoid plant hormones. Nature 455, 195-200, doi:10.1038 / nature07272 (2008). 9. Lopez-Obando, M., Ligerot, Y., Bonhomme, S., Boyer, F.-D. & Rameau, C. Strigolactone biosynthesis and signaling in plant development. Development 142, 3615-3619, doi: 10.1242 / dev. 120006 (2015). 10. Daignan Fornier, S. Total synthesis of cannalactone, non-canonical strigolactone from hemp and development of synthetic analogues for their biological evaluation. Université Paris-Saclay, (2023). 11. Daignan Fornier, S., de Saint Germain, A., Retailleau, P., Pillot, J.-P., Taulera, Q., Andna, L., Miesch, L., Rochange, S., Pouvreau, J.-B. & Boyer, F.-D. Noncanonical Strigolactone Analogues Highlight Selectivity for Stimulating Germination in Two Phelipanche ramosa Populations. J. Nat. Prod. 85, 1976-1992, doi:10.1021 / acs.jnatprod.2c00282 (2022). 12. Macalpine, G.A., Raphaël, R.A., Shaw, A., Taylor, A.W. & Wild, H.J. Synthesis of Germination Stimulant (±)-Strigol. J. Chem. Soc., Perkin Trans. 1, 410-416, doi:10.1039 / C39740000834 (1976). 13. Boutet-Mercey, S., Perreau, F., Roux, A., Clavé, G., Pillot, J.-P., Schmitz-Afonso, I., Touboul, D., Mouille, G., Rameau, C. & Boyer, F.-D. Validated Method for Strigolactone Quantification by Ultra High-Performance Liquid Chromatography - Electrospray Ionization Tandem Mass Spectrometry Using Novel Deuterium Labeled Standards. Phytochem. Anal. 29, 59-68, doi:10.1002 / pca.2714 (2018). 14. Jas, G. A Simple Resolution of 4-Bromo-2-(Tert-Butyldimethylsiloxy) Furan from Tetrahydro-2,4-Dioxofuran. Synthesis 11, 965-966. doi:10.1055 / s-1991-26618 (1991).

Claims

Claims

1. Deuterium-labeled cannalactone, characterized in that it conforms to the general formula (1): [Chem.l] / S ■W "R8 in which: - R1 denotes a hydrogen atom H or a deuterium atom D, - R2 denotes the methyl radical CH3 or the tri-substituted deuterated methyl radical CD3, - at least one of R1 and R2 being deuterated.

2. Deuterium-labeled cannalactone according to claim 1, in which: - R1 denotes H, and - R2 denotes CD3.

3. Deuterium-labeled cannalactone according to claim 1, in which: - R1 denotes D, and - R2 denotes CH3.

4. Deuterium-labeled cannalactone according to claim 1, in which: - R1 denotes D, and - R2 denotes CD3. [Claim] 5] Process for the synthesis of cannalactone, of general formula (5) [Chem. 5] -Vfeç.fe9 Ta characterized in that it comprises the following steps: - a reaction A) coupling of commercial B-cyclocitral with a C4 bromofuran of formula (6) [Chem.6] 1 >» <WS. to obtain a B20 alcohol of formula (7) [Chem.7] - a step B) of reduction of alcohol B20 of formula (7), to obtain a mixture of diastereomers of allylic alcohol, followed by a step of separation, of said diastereomers to retain the diastereomer (4R*, 6R*)-B21 of formula (8) [Chem. 8] (4RA,} Bit - a step C) of selective epoxidation of the compound (4R*, 6R*)-B21 of formula (8) to obtain the 4R*, 6R*)-cis-B22 epoxide of formula (9) [Chem.9] (4R\ - a radical reaction D) of reduction and isomerization of the (4R*, 6R*)-cis-B22 epoxide of formula (9), to obtain the tertiary alcohol (4R*, 8R*)-B1 of formula (10) [Chem. 10] pjr, 8«>EH - a step E) of protection of the tertiary alcohol (4R*, 8R*)- B1 of formula (10), to obtain the protected alcohol (4R*, 8R*)-B23 of formula (11) [Chem. 11] - a step F) comprising the formylation Fl) of the compound (4R*, 5R*)-B23 of formula (11) using alkyl formate corresponding to formula (12) [Chem. 12] then the O-alkylation F2) of the aldehyde thus formed using the compound D4 of formula (13) [Chem. 13] -... / 'SA 04 leading to the obtaining of the mixture of diastereomers (47?*,67?*)-B24 of formula (14) [Chem. 14] J'otiwVj (4R'r 8^)624 - a step G) of deprotection and separation of the diastereomers of formula (14), leading to the isolation of cannalactone of formula (5).

6. A process for synthesizing a deuterium-labeled cannalactone as defined in any one of claims 1 to 4, characterized in that it comprises the following steps: - steps A to E of the synthesis of the protected compound (4R*, &7?*)-B23 of formula (11) as defined in claim 5; - a step F) comprising the formylation F1) of the compound (47?*, 87?*)-B23 of formula (11) using ethyl formate corresponding to formula (12) or using deuterated alkyl formate corresponding to formula (15) [Chem. 15] o - then the O-alkylation of the aldehyde thus formed using compound D4 of formula (13) [Chem. 13] 04 or using the deuterated compound D4 of formula [Chem. 16] w at least one of the organic compound functionalized by a formyl group or of the D4 compound being deuterated, to obtain a mixture of deuterated diastereomers (4R* 67?*)-B24 of formula (17) end \ 0 ï & -0 1 with - R1 denoting a hydrogen atom H or a deuterium atom D, and - R2 denotes a methyl group (CH3) or a methyl group where the hydrogens have been replaced by deuterium atoms (CD3); - a step G) of deprotection and separation of the diastereomers, leading to the isolation of a deuterium-labeled cannalactone of formula (1).

7. A process according to claim 6, wherein: - the formylation Fl) is carried out on the alkyl formate corresponding to formula (12); and - the O-alkylation F2) of the aldehyde following the formylation Fl) is carried out using compound D4 of formula 16.

8. A process according to claim 6, wherein: - the formylation Fl) is carried out using the deuterated alkyl formate corresponding to formula (15); and - the O-alkylation F2) of the aldehyde following the formylation Fl) is carried out using compound D4 of formula (13).

9. The method according to claim 6, wherein: - the formylation Fl) is carried out using the organic compound functionalized with a formyl group corresponding to formula (15); and

10.

11. - the O-alkylation F2) of the aldehyde following the formylation Fl) is carried out using compound D4 of formula (16). Use of deuterium-labeled cannalactone as defined in any one of claims 1 to 4 or as obtained in any one of claims 6 to 9, for the determination of cannalactone in any tissue or exudate of plants or living organisms, and in particular in the exudates or tissues of the hemp plant. Use according to claim 10, for determining the natural cannalactone content of hemp plant exudates or tissues by liquid chromatography coupled with mass spectrometry.

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