DD147239A1 - METHOD FOR PRODUCING AZIDOGIBBERELLINES - Google Patents

Abstract

It is the object of the invention to prepare such gibberellin derivatives which have an altered biological activity and in particular contain a grouping suitable for the photoaffinity labeling technique. The object of the invention is to introduce an azido group into position 1 of the ent-gibberellan skeleton. According to the invention, a gibberellin derivative having an * & exp1! -3-oxo structural feature is reacted with hydrazoic acid in an inert solvent and optionally subsequently reduced with complex metal hydrides, in particular with sodium borohydride. 1-Azidogibberellins are already obtained in good yields when working at room temperature. By-products were not found.

Description

21.6763 - "-

Process for the preparation of azidogibberellins

The invention relates to a process for the preparation of Azidogibberellinen. These are gibberellin derivatives which have an azido group in position 1 of the ent-gibberellan skeleton. The ent-gibberellan skeleton is intact or slightly modified (for example, seco, homo or nor-ent gibberellan) and optionally substituted in various ways.

Field of application of the invention

The field of application of the invention is biochemical research, in particular the investigation of the regulation of growth and differentiation processes in plants with regard to the development of new growth regulators for agricultural practice.

The azidogibberellins contain a chromophore function suitable for the photoaffinity labeling technique and can thus be used for receptor studies as phytohormone analogs.

The compounds prepared according to the invention also serve to synthesize further structure-modified gibberellins or as starting materials for the expression of carrier-fixed gibberellins for affinity chromatography.

Characteristic of the known technical solutions

Naturally occurring diterpenoids with a Gibban skeleton possess as phytohormones of multiple action (gibberellins) large

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Biological significance. It is known that structural modification of such gibberellins leads to compounds with altered biological activity. For example, deoxygibberellin C, pseudogibberellin A ^ and certain fluorinated species exhibit gibberellin-antagonistic properties «

Numerous other structurally modified gibberellins have been prepared by partial or total synthetic methods and tested for their biological activity. So z. B. by acid-catalyzed hydration of 3-dehydro-gibberellin A, with 2 η hydrochloric acid in tetrahydrofuran and subsequent reduction with sodium borohydride 1ß-hydroxy-gibberellin A ^ and 1ß-hydroxy-pseudogibberellin A ^ synthesized (G. ADAM and Ph. HUNG, DDR patent 112 439, C 0? D, 5 / 06j G. ADAM and Ph. D. HUNG, Tetrahedron Letters 1974, 3419) and by photochemical or alkali-catalyzed reaction of the 3-dehydro-gibberellin A, methyl ester in alcohols 1-alkoxy-gibberellin-1-methyl ester (EP SEREBRYAKOV, NS KOBRINA, VF KUCIIEROV, G. ADAM and K. SCHREIBER, Tetrahedron 28 (1972), 3819; IA GURVICH, IM MILSTEIN and VP KQCHE ¹ ROV, Izvest. Akad. Nauk SSSR (Ser. Khim.) 1966, 184). Gibbere11in compounds which have an unsubstituted or substituted mercapto group at position 1 of the ent-gibberellan backbone were prepared by reacting gibberellin derivatives having a -3-oxo structural feature with a compound R-SH, wherein R is H, alkyl or Substituted alkyl is prepared in a solvent and optionally in the presence of an organic base (B. VOIGT and G. ADAM, DDR Patent 136 836, C 07 D 307/93).

Object of the invention

It is the object of the invention to obtain gibberellin derivatives having altered biological activity, in particular to those gibberellin derivatives containing a grouping suitable for the photoaffinity labeling technique. ,

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Explanation of the essence of the invention

The object of the invention is to produce gibberellin derivatives which have an azido group in position 1 of the ent-gibberellan skeleton.

According to the invention, a gibberellin derivative having an A 2-oxo structural feature is reacted with hydrazoic acid and optionally subsequently reduced with complex metal hydrides.

The hydrazoic acid is used in an inert organic solvent, for example in ethereal solution.

Suitable starting materials gibberellin derivatives include 3-dehydro-gibberellin A ^, 3-dehydro-gibberellin A ₁ - ,, 3-Dehydrogibberellin A, q, 3-dehydro-gibberellin A ^ p or the corresponding methyl esters. They are prepared by known methods (see, for example, BPJ KEAY, JS MOFFATT and TPC MULHOLLAKD, J. ehem. Soc.1805 (1965) 5 NS KOBRINA, EP SEREBRYAKOV, VP KÜCHEROV, G. ADAM and B. VOIGT, Tetrahedron 2 ^ J425 (1973)).

The reaction is carried out in an inert solvent, e.g. For example, tetrahydrofuran, diethyl ether or methylene chloride performed. 1-Azidogibberellins are already obtained in good yields when working at room temperature. By-products were not found. ,

In order to improve stability, water solubility and absorption and transport of the compounds in the plant, it is expedient to reduce the keto group in position 3 with complex metal hydrides, in particular sodium borohydride.

The isolation of Azidogibberelline prepared by known methods.

The azidogibberellins produced according to the invention have growth-active substances of biological interest and practical significance for photoaffinity labeling studies.

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Even after reduction of the keto group in position 3, a gibberell effect is retained; if necessary, it is even intensified.

The invention will be explained in more detail by the following exemplary embodiments:

embodiments Example 1:

1β-Azido-3-dehydro-gibberellin A ₁ (3) and 1 # -azido-3-dehydrogibberellin A ₁ (4):

280 mg of 3-dehydro-gibberellin A ^ (1) were dissolved in 30 ml of absolute. Tetrahydrofuran with 5 ml of an ethereal solution of hydrazoic acid (from 1 g NaN.,) And 2 days at room temp. left to stand »After distilling off the solvent i. Vak. were 317 mg of an amorphous mixture of 3 and 4 in the ratio 2.8 ι 1 obtained. IR (nujol): ^ max 908 OC = CH ₂ ), 1703 and 1719 (carbonyl), 1782 (f-lactone), 2103 and 2132 (azide) and 3408 cm ^-1 "(hydroxyl). UV (CH ₃ OH, c = 0.937): TSnax 286 nm, E _M = 259. MS: m / e 3 ^ 4 (M ⁺ -HNO ₀ NMR: 1.09 (s, 18-H _$ ), 1, 11 (s, 18-H ₃ ), 2.27 (m, 2-H ₂ ), 2.76 (d, J = 9 Hz, 6-H), 3.30 (d, J = 9 Hz, 5 -H) ₁ 4.45 (dd, J ₁ = .8.5 Hz, J ₂ = 2.5 Hz, 1β H), 4.71 (dd, J ₁ = 6.5 Hz, J ₂ = 1 , 5 Hz, KH), 4.95 and 5.25 (<T) ppm (17 -H ₂ ).

Example 2:

1β-azido-3α ⁽ i6b.-hydrogibberellin A ₁ -methylester (5) and 1α-azido-3-dehydro-gibberellin A ₁ -methylester (6):

1 * 074 g of 3-dehydro-gibberellin A, methyl ester ("2) were reacted in 250 ml of methanol-free methylene chloride with 25 ml of an ethereal solution of hydrazoic acid (from 3 g of NaN 2) at room temp. After distilling off the solvent. In a vacuum, 1.20 g of an amorphous mixture of 5 and 6 were obtained in a ratio of 1.8: 1, IR: max max 907), 1176 (ester carbonyl), 1723 (carbonyl), 1779

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(y-lactone), 2102 (azide), 3078 (rc = CH ₂ ) and 3400 cm ^-1

y ₂

(Hydroxyl). UV (c = 0.910): inax 290 ma, E ₁ 267. MS: m / e 358 (M ⁺ -HU ₅ ). MMR: 1.02 (s, 18-H,), 1.06 (s, 18-H,), 2.15 (m, 2-H ₂ X, 2.81 (d, J = .9 Hz, 6-H), 3.51 (d, J = 9 Hz, 5-H), 3.72 (s, COOCH ₃ ), 4.30 (ad, J ₁ = 9 Hz, J ₂ = 2.5 Hz , I-H), 4.72 (dd, J ₁ = 7 Hz, J ₂ = 2 Hz, '1A.-H), 4.96 and 5.28 (if) ppm (17-H ₂ ).

Example 3? IB-azido-gibberellin A ₁ (7):

1.133 g of 3 and 4 were in 200 ml of methanol with 550 mg of sodium borohydride within 30 minutes with stirring at room temp. reduced, after acidification with 10 ml of 20% acetic acid, the solvent was i. Vak. largely distilled off, the remaining solution is mixed with 50 ml of water and extracted 10 times with 25 ml of ethyl acetate. The combined ethyl acetate extracts were washed with water, dried over sodium sulfate and i. Vak. evaporated: 1.27 g of crude product, which was bound to 2.5 g of Celite and on 62 g of silica gel (Woelm) was chromatographed (30 ml fractions). Elution with benzene / ether 4: 6 ^v / v gave in fractions 36-49 509 mg (40%) 7 of mp ⁰ C (Zers from acetone / n-hexane) and ϋϊΐψ 625 °

239-41 ⁰ C (dec, from acetone / n-hexane.) And ϋϊ-ΐψ 62.5

(c = 0.160, C ₂ H ₅ OH). IR: ^ max 906 (^ C = CH ₂ ), 1704 (carbonyl),

1762 (^ -lactone), 2113 (azide) and 3 ^ 53 cm * " ¹ (hydroxyl). MS:

m / e 389 (M ⁺ ). MR (100 MHz): 1.07 (s, 18-H,), 2.56 (d, J = 10 Hz, 6-H), 3.52 (d, J = 10 Hz, 5-H), 3.76 (dd, J ₁ = 5 Hz, J ₂ = 2 Hz, 3 H), 4.08 (dd, J ₁ = 5 Hz, J _£ s 2 Hz, 1 H), 4.86% and 5.16 (<f) ppm (17H ₂ ) *

1β-azido-pseudogibberellin A ₁ (8):

Elution with benzene / ether 2: 8 ^v / v of the column chromatography described for 7 afforded 5O-83 296 mg in the fractions (23% amorphous 8 with Ctj ^ - 11.9 ° (c = 0.311).) IR: ^ max C C s CH ₂ ), 1707 (carbonyl), 1763

2102 (azide), 3078 (^ C = CH ₂ ) and 3 ^ 20 cm ^-1 "(hydroxyl). MS: m / e 389 (M ⁺ ). MR (100i.Sz): 1.11 (s, 18-H,), 2.55 (d, J =

-ΟΙΟ Hz, 6-H), 2.69 (d, J · = 10 Hz, 5-H), 3.79 (dd, J ₁ = 12 Hz, J ₂ = 6 Hz, 3-H), 4 , 17 (dd, J ₁ = 6 Hz, J ₂ = 2 Hz, 1-H), 4.85 and 5.18 (<f) ppm

1 <t-azido-gibberellin A ₁ (9) J

Further elution with ether / glacial acetic acid 98: 2 ^v / v gave in the fractions 95 - 115 306 mg (24%) amorphous 9 with Zt7p ² + 45.3 ^O (c = 0.256). IR: ^ max 907 OC = CH ₂ ), 1706 (carbonyl), 1762 Q'-lactone), 2101 (azide) and 3400 cm ^-1 "(broad, hydroxyl). MSi m / θ x 389 (M ⁺ ). IiMR (100 MHz): 1.08 (s, 18-H ₃ ), 2.54 (d, J s 10 Hz, 6-H), 3.53 (d, J = 10 Hz, 5-H), 3, 64 (t, J = 5 Hz,

5-H), 3.73 (dd, J ₁ = 7 Hz, J ₂ = 4 Hz, 1-H), 4.80 and 5.17 ppm

The following were prepared analogously: 1β-azido-gibberellin A ₁ -methyl ester (10): mp 193-95 ° C (needles from acetone / n-hexane) / 7 ^ / ψ -66.9 ° (c = 0.329). IR: ^ max 907 OC = CH ₂ ), 1177 (ester carbonyl), 1657 OC = CH ₂ ), 1733 (carbonyl), 1758 (^ -lactone), 2118 (azide), 3080 OC = CH ₂ ), 3420 and 3460 cm ^-1 (hydroxyl) MS: m / e 402 (M ⁺ -I) MIR (100 MHz): 1.06 (s, 18-H,), 2.60 (d, J s 10 Hz, 6-H), 3.53 (d, J = 10 Hz, 5-H), 3.66 (s, COOCH ₃ ), 3.78 (3-H), 4.07 (dd, J ₁ = 5 Hz, J ₂ = 2 Hz, 1-H), 4.72 (d, J = 4.5 Hz, H-bridge), 4.83 and 5.14 (<ί) ppm (17-H ₂ ). Yield: 39 %.

1β-azido-pseudogibberellin Αχ, -methyl ester (11): amorphous, ßij ^ - 10,2 ° (c = 0,304). IRpmax 906 (> C =

^ ₂

1174 (ester carbonyl), 1655 Oc = CH ₂ ), 1725 (ester carbonyl), 1772 ττ lactone), 2098 (azide), 3073 O C = CH ₂ ) and 3420 cm ^-1 (hydroxyl). MS: m / e 403. (M ⁺ ). MR: 1.05 (s, 18-H _$ ), 2.63 (d, J = 10 Hz, 6-H), 2.88 (d, J = 10 Hz, 5 H), 3.71 (s, COOCH ₃ ), 3.90 (3-H), 4.21 (dd, J ₁ = 5 Hz, J ₂ = 2 Hz, 1-H), 4.91 and 5 , 21 U) ppm (17H ₂ )

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Claims (2)

-? - 2Ί έ claim for invention, 1. A process for the preparation of Azidogibberellinen, characterized characterized in that a gibberellin derivative having A -J-oxo structural feature is reacted with hydrazoic acid in an inert solvent and optionally subsequently reduced with complex metal hydrides. 2. The method according to item 1, characterized in that after the implementation of the gibberellin derivative with Δ. 3-oxo structural feature is reduced with hydrazoic acid with sodium borohydride.