JP2564618B2 - Process for producing 3 ', 4'-anhydrovinblastine - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a novel method for efficiently producing 3 ', 4'-anhydrovinblastine.

The compound obtained by the present invention itself has useful physiological activities such as antitumor activity, and is also useful as a raw material for producing vinblastine and vincristine known as anticancer agents.

[Conventional technology]

3 ', 4'-anhydrovinblastine brass Chin (Δ ^15,20 -Anh
ydrovinblastine) is a known compound with antitumor activity (US Pat. No. 4,029,663).

3 ′, 4′-anhydrovinblastine is conventionally obtained by extracting it from Catharanthus roseus (also known as Vinca rosea.L), which is a plant of the genus Periwinkle, or chemically converting casalanthin and bindrin obtained from Catharathus roseus. Can be obtained by reacting to.

However, the extraction method requires a very small amount in the plant, and a large number of similar compounds exist in the plant, which makes separation and purification difficult.

On the other hand, as a chemical synthesis method, casalanthin is participated by peracid, the produced N-oxide is reacted with vindoline after acylation, and then reduced with NaBH ₄ or the like (US Pat. No. 4,144,237), and casalanthine and Feldbindolin
A method is known in which coupling is performed in the presence of ³⁺ and then reduction is performed with NaBH ₄ . However, in the former method,
Isolation yield of 3 ', 4'-anhydrovinblastine is 41%
Is low, and 10% of isomers are produced at the same time, so separation and purification are difficult. In addition, although the latter method shows a high value of 68.6% even though it is a liquid chromatography yield, this reaction also produces various compounds such as 12.3% vinblastine at the same time. Difficult.

[Problems to be Solved by the Invention]

It is an object of the present invention to further improve the yield of a method for producing 3 ', 4'-anhydrovinblastine by reacting casalanthin with vindoline in the presence of Fe3 ⁺ .

[Means for solving the problem]

The present invention has been made in order to achieve the above-mentioned object, and among the above-mentioned known methods, from the industrial viewpoint, catharanthine and vindoline.
Attention was paid to the method of coupling after the coupling in the presence of Fe ³⁺ and then reducing. As a result of examining from various directions, we paid attention to Fe ³⁺ , which was conventionally considered to be harmless even if left in the reaction system after the coupling reaction, and carried out the reduction reaction after removing Fe ^3+. However, contrary to what is expected from conventional common knowledge, not only the yield of the target substance increases significantly, but also complicated physical purification means such as chromatographic treatment is required for separation and recovery of the target substance. We have discovered an unexpected effect, which is particularly suitable for industrial methods, since it can be directly crystallized without being separated and easily separated and recovered.

As a result of further research based on this new knowledge, the present invention has been completed. That is, the first invention of the present invention is
After the Kasaranchin and vindoline were reacted in the presence of Fe ^3+, a method to inactivate the removal of Fe ³⁺ or Fe ^3+, followed by reduction 3 ', to produce a 4'-anhydrovinblastine Chin It is in. And the second invention is the above method, wherein Fe ³⁺ is inactivated by adding an iron ligand,
It is a method for producing 3 ', 4'-anhydrovinblastine.

In carrying out the present invention, the reaction between casalanthin and vindoline in the presence of Fe ³⁺ may be carried out as usual.

After the completion of this coupling reaction, Fe ³⁺ is removed or an inactivation treatment is carried ^out.For the removal of Fe ³⁺ , a basic compound is added after the completion of the coupling reaction to precipitate Fe ³⁺ . It is produced and removed from the system by a commonly used solid-liquid separation method such as filtration or centrifugation. The basic compound used herein refers to all basic compounds capable of forming Fe ³⁺ precipitates without interfering with the reaction, and examples thereof include the following: Metal hydroxides such as sodium, potassium hydroxide, calcium hydroxide and barium hydroxide, carbonates such as sodium carbonate, potassium carbonate and ammonium carbonate, bicarbonates such as sodium bicarbonate and potassium bicarbonate, urea and methyl Amines such as amine, ethylamine, dimethylamine and trimethylamine, quaternary ammonium hydroxide such as tetraethylammonium hydroxide, and ammonia water.

The Fe ³⁺ passivation treatment broadly means a treatment for removing or reducing the adverse effect or interfering action of Fe ³⁺ in the subsequent step.

Therefore, the inactivation treatment of Fe ³⁺ widely includes a treatment for suppressing the coordination of iron with a useful component in the subsequent step or an action for suppressing the oxidation action of iron, for example, Fe ³⁺ In order to eliminate the adverse effects or interference of Fe ³⁺ by adding an iron ligand after completion of coupling to reduce Fe ³⁺ and reduce Fe ³⁺ to metallic iron or Fe ^2+. Methods of activation, methods of reacting different kinds of ligands to form different kinds of ligands respectively, and then making these complexes into double salts to coprecipitate or non-toxic melts, etc. are widely included. It In addition, the method of leaving the precipitate of Fe ^{3+ with} a basic compound in the system without removing it at all, or removing a part of the precipitate and leaving the other residue, is also suitable for inactivation. This is an example of the method.

Among the Fe ³⁺ deactivation treatments, the method of deactivating by adding an iron ligand is a method of adding a compound that coordinates with an iron ligand, that is, Fe ³⁺ . The following compounds are listed as the iron ligand used in the present invention, but other complexing agents can also be widely used: Lower fatty acids: formic acid, acetic acid, propionic acid, butyric acid, valeric acid, trimethylacetic acid, Caproic acid, enanthic acid, caprylic acid, etc.

Dicarboxylic acid: Aliphatic dicarboxylic acid (oxalic acid, malonic acid, succinic acid, glutaric acid, adipic acid, maleic acid,
Fumaric acid, pimelic acid, etc.); aromatic dicarboxylic acids (phthalic acid, isophthalic acid, terephthalic acid, etc.), etc.

Ketocarboxylic acid: pyruvic acid Hydroxycarboxylic acid: Aliphatic hydroxycarboxylic acid (glycolic acid, lactic acid, hydroallylic acid, glyceric acid, malic acid, tartaric acid, succinic acid, citric acid, etc.); Aromatic oxycarboxylic acid (salicylic acid, oxybenzoic acid) , Gallic acid, etc.) etc.

Diol: ethylene glycol, catechol ascorbic acid, etc.

Polyaminocarboxylic acid: EDTA, etc.

Sugar derivative: glucose, fructose, sucrose, arbutin, etc.

These salts: Cyanide: Potassium cyanide, Sodium cyanide,
Tributylammonium cyanide, etc.

Thiocyanide: potassium thiocyanate, sodium thiocyanate, thiocyanated tributylammonium, etc.

Fluoride: Sodium fluoride, potassium fluoride, etc.

Pyridine derivative: phenanthrene derivative, dipyridyl derivative and the like.

Phosphoric acid and condensed phosphoric acid: polyphosphoric acid, metaphosphoric acid, pyrophosphoric acid, phosphoric acid and salts thereof.

These iron ligands are 0.5 to 200 times mol relative to Fe ³⁺ ,
It is preferable to add 1 to 10 times mol.

After inactivating Fe ³⁺ in this way, a reducing agent is added to the reaction system to form the target compound.

If Fe ³⁺ precipitates due to the inactivation treatment,
The reaction liquid such as the filtrate after removing the precipitate may be subjected to the above treatment. On the other hand, the removed precipitate is washed with an organic solvent, and then this washing liquid is combined with the reaction liquid such as the filtrate or the supernatant,
After concentrating under reduced pressure to remove the organic solvent and adding water again, NaBH ₄
PH <6 without reducing with a reducing agent such as
After that, the liquid may be separated and reduced by adding a reducing agent such as NaBH ₄ to the water tank. In addition, the precipitate generated by the inactivation treatment was obtained by extracting the aqueous layer with an organic solvent as it is without removing it, or when the inactivation treatment was performed in a liquid state, without performing anything. There is a method in which the obtained organic layer is concentrated under reduced pressure, water is added, and then reduction is performed.

Examples of the reducing agent include, but are not limited to, hydrogen compounds: sodium borohydride, potassium borohydride, lithium borohydride, sodium cyanoborohydride; metals: zinc, iron, tin, aluminum, magnesium;
And others.

When the reducing power is strong, these reducing agents may be used in a reduced amount or may be modified or masked.

The coupling reaction may be carried out in an air atmosphere, but better results can be obtained if it is carried out in a nitrogen atmosphere or under aeration of nitrogen.

The 3 ', 4'-anhydrovinblastine produced by these methods can be obtained by extraction with an organic solvent.

According to this method, since the target compound has a very high purity,
It can be crystallized freely, and it is possible to sufficiently recover it by recrystallization without the need for advance separation and purification such as column treatment, which was essential in the case of a low-yield conventional method. Thus, this method is very suitable as an industrial method because of its easy separation.

〔The invention's effect〕

According to the method of the present invention, the yield of 3 ', 4'-anhydrovinblastine was 68.6% by liquid chromatography.
% To 89%, and 3 ', 4'-anhydrovinblastine can be obtained as highly pure crystals without the need for separation and purification.

〔Example〕

Next, the present invention will be described in more detail by way of examples, which should not be construed as limiting the present invention.

Example 1 180 ml of water was added to a 500 ml reaction vessel and cooled with ice while blowing N ₂ gas. There, 1/2 sulfate of casalanthin 184mg
(480 μmol) dissolved in 20 ml of water and Bindrin 22
A solution prepared by dissolving 0 mg (480 μmol) in 20 ml of water and 0.4 ml of 2N-HCl was added. A solution prepared by dissolving 8.1 g of FeCl ₃ .6H ₂ O in 50 ml of water is added thereto to start the reaction. While blowing N ₂ ,
After stirring under ice cooling for 3 hours, a solution of 12.2 g of NaOAc dissolved in 50 ml of water is added. To it, 227 mg of NaBH ₄ was added as a solid,
Stir for 30 minutes. The reaction mixture is extracted 4 times with 100 ml of ethyl acetate, the organic layer is dried over anhydrous sodium sulfate and then concentrated under reduced pressure. After dissolving the obtained residue in 30 ml of chloroform,
Wash with 50 ml of saturated aqueous NaHCO ₃ . The organic layer is dried over anhydrous sodium sulfate and then concentrated. Crystals are formed when 2 ml of methanol is added to the residue. The produced crystals were filtered to obtain 304 mg of colorless crystals of 3 ', 4'-anhydrovinblastine having an mp of 200 to 204 (decomposition) ° C. The product yield was 80%. This compound was 3.89 (3H, S) at ¹ H-nmr (CDCl ₃ ).
It gives a signal to 3.54 (3H, S) and is a natural 3 ', 4'-anhydrovinblastine.

Example 2 Water (10 ml) was placed in a 50 ml reaction vessel and bubbled with nitrogen for 30 minutes while cooling with ice. After that, 11.9 mM casalanthin 1/2 sulfate aqueous solution 20 μ (2.39 μmol) and 12.0 mM hysidrin hydrochloric acid solution 20 μ (2.41) were added. μmol) and 1.0 M (1.20 mmol) of 1.2 M ferric chloride aqueous solution are added in this order, and the mixture is stirred for 3 hours while bubbling nitrogen under ice cooling. The mixture is made alkaline with 1 ml of 25% aqueous ammonia and extracted three times with 10 ml of ethyl acetate. The extracts were combined and dried under reduced pressure, then water (10m
l), dissolved in 2N-HCl (24μ), added with 1 ml (227μmol) of 227mM aqueous solution of sodium borohydride, and stirred for 30 minutes.
Then, add 2 ml of 25% ammonia water, and add 10m ethyl acetate.
Extract 3 times with l.

The extracts were combined, dried under reduced pressure at 40 ° C. or lower, and analyzed by high performance liquid chromatography under the following conditions. As a result, the yield of 3 ', 4'-anhydrovinblastine was 89%.
Got with.

Column: YMC-Packed column AM-312 (S-5 120AODS) Solvent: CH ₃ CN: 0.01M ammonium carbonate aqueous solution = 3: 2 Flow rate: 1 ml / min Column temperature: 45 ° C Detection wavelength: 254 nm Retention time: 3 ', 4'-Anhydrovinblastine (40.8 minutes) Example 3 In Example 2, 1 ml of 25% aqueous ammonia was used, and L
After the addition of 634 mg (3.6 mmol) of ascorbic acid, 3 ml of the same treatment as in Example 2 was performed except that 1 ml (227 μmol) of a 227 mM aqueous solution of sodium borohydride was added and the mixture was stirred for 30 minutes.
4'-Anhydrovinblastine was obtained with a yield of 88%.

Example 4 The same procedure as in Example 3 was carried out except that L-ascorbic acid was replaced by triammonium citrate, and the yield of 3 ', 4'-anhydrovinblastine was 87%.
Got with.

Example 5 3 ', 4'-anhydrovinblastine was obtained in a yield of 85% by performing the same treatment as in Example 3 except that sodium pyruvate was added instead of L-ascorbic acid in Example 3.

Example 6 The same treatment as in Example 3 was performed except that oxalic acid was added instead of L-ascorbic acid in Example 3, and 3 ′, 4 ′.
-Anhydrovinblastine was obtained with a yield of 87%.

Example 7 The same treatment as in Example 3 was performed except that malic acid was added in place of L-ascorbic acid in Example 3 to obtain 3 ', 4'.
-Anhydrovinblastine was obtained with a yield of 88%.

Example 8 The same treatment as in Example 3 was carried out except that maleic acid was added instead of L-ascorbic acid in Example 3, and 3 ′,
4'-Anhydrovinblastine was obtained with a yield of 84%.

Example 9 3 ', 4'-anhydrovinblastine was obtained in a yield of 85% by performing the same treatment as in Example 3 except that sodium fluoride was added instead of L-ascorbic acid.

Example 10 The same treatment as in Example 3 was performed except that arbutin was added in place of L-ascorbic acid in Example 3, and 3 ′,
4'-Anhydrovinblastine was obtained with a yield of 82%.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Takenori Takechiro 6-1-2 Waki, Waki-cho, Kuga-gun, Yamaguchi Prefecture Mitsui Petrochemical Industries, Ltd. (72) Noriaki Kihara Waki, Waki-cho, Kaku-gun, Yamaguchi Prefecture 6-1-2, Mitsui Petrochemical Industry Co., Ltd.

Claims (2)

(57) [Claims] After 1. A a Kasaranchin and vindoline were reacted in the presence of Fe ^3+, 3 to inactivate the removal of Fe ³⁺ or Fe ^3+, and then characterized by reduced ', 4' -A method for producing anhydrovinblastine. 2. The 3 ', 4'- according to claim 1, characterized in that Fe ³⁺ is inactivated by adding an iron ligand.
A method for producing anhydrovinblastine.