JPH0138104B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel method for producing prostaglandin derivatives. For details, see the formula 5,8,9,11,12,14,15-heptadieutero-6-oxo-prostaglandin
This relates to the manufacturing method of F ₁ 〓.

The compound of formula () obtained by the present invention is
6-oxo- which is a decomposition composition of PGI ₂ ()
It is the 5,8,9,11,12,14,15-heptadieutero form of PGF ₁ 〓 (), and the amount of PGI ₂ () in the blood is determined by its degradation product 6-oxo PGF ₁ 〓 (). It is useful as an internal standard reagent for quantitative determination.

PGI ₂ () is synthesized in the arterial wall and has a wide range of physiological actions, including strongly inhibiting platelet aggregation, suppressing gastric acid secretion, and dilating peripheral blood vessels. The production amount changes depending on various diseases, so the amount of production in the body changes.
Measuring the amount of PGI ₂ () is expected to be useful in diagnosing diseases. However, PGI ₂ is extremely unstable and breaks down quickly even if it is produced in the body, making it difficult to directly measure the amount of PGI ₂ . Therefore, this measurement is currently carried out by quantifying 6-oxoPGF ₁ 〓(), which is a decomposition product of PGI ₂ .

However, PGI ₂ (6-oxo
Since PGF ₁ 〓) is in a very small amount, its measurement requires nano to picogram levels. Therefore, gas chromatograph-direct mass spectrometry, which is said to have the highest detection sensitivity, is used for such measurements. In this case, a deuterium labeled substance is selected as the internal standard, which has the same chemical properties, retention time in gas chromatography, and cleavage mechanism in mass spectrum as the substance to be measured, and differs only in fragment ions. In order to further reduce errors, the number of deuterium atoms is determined so that the peak of the internal standard and the peak of the substance to be measured do not overlap on the mass spectrum chart. In the mass spectrum of ordinary organic compounds, the main peaks +1, +
Since peaks based on heavy isotopes appear in 2,
In order to prevent peaks from overlapping, the number of deuterium atoms in the internal standard compound must be at least 3 or more. Further, since the measurement becomes easier when both peaks are clearly separated, a compound having 6 to 7 deuterium atoms is ideal as an internal standard.

As mentioned above, 6-, which has 6 to 7 deuterium atoms,
If gas chromatograph direct mass spectrometry is performed using oxoFPG ₁ 〓 as an internal standard, PGI ₂ (6-
It is expected that accurate quantification of oxo-PGF ₁ 〓) will become easier and will be useful in diagnosing diseases.

By the way, 6-oxoPGF ₁ 〓, the compound (), which has 7 deuterium atoms, has already been synthesized by an enzymatic method (C. Pace-Asia, J.
Amer.Chem.SOC., 98, 2348 (1976)) However, the enzymatic method is not suitable for economically synthesizing compound () in large quantities because enzymes derived from pig aortic microsomes are difficult to obtain. On the other hand, 5, 6, 8, 9,
11, 12, 14, 15-Octage Utero-PGF ₂ 〓()
5, 6, 8, 11, 12, 14, 15-heptage
Terrorism-PGE ₂ is 5, 6, 8, 9, 11, 12, 14,
It is known that it can be synthesized in large quantities from 15-octadiutelo-arachidonic acid using an enzyme derived from sheep seminal glands. Therefore, as a result of intensive study on a method for producing compound () by an organic chemical method using () obtained in large quantities by an enzymatic method as a starting material, a new industrial and inexpensive production method was discovered and the present invention was completed. This is what I did.

The present invention will be described in detail below.

Compound (ii) of the present invention has the general formula () [In the formula, R represents a tetraalkylammonium substituted with an alkali metal, an alkaline earth metal, or an alkyl having 1 to 4 carbon atoms.] It can be obtained by hydrating a compound represented by the following with an acid. That is, it can be obtained in an almost pure form by dissolving the compound of general formula () in a solvent, adjusting the pH using various acids, extracting with an organic solvent, and concentrating the solvent. Generally, water is preferably used as the solvent for this purpose, but an organic solvent may also be present in the coexistence. Examples of organic solvents that can coexist include ether, 1,2-dimethoxyethane, dioxane, DMF, DMSO, ethyl acetate, benzene, toluene, chloroform, etc., but are not limited thereto. Examples of acids include organic acids such as oxalic acid, acetic acid, trifluoroacetic acid, and P-toluenesulfonic acid, and inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, and phosphoric acid. Among these, oxalic acid and hydrochloric acid are preferred, but of course the acid is not limited to these.

PH is preferably 4 to 2, particularly preferably 3 to 3.5.
Extraction solvents include those commonly used for extraction of organic compounds, such as ethyl acetate, toluene, benzene, chloroform, and ether. Among them, ethyl acetate is preferably used, but of course it is not limited to this. The reaction temperature is preferably -10° to 25°C, particularly preferably 0° to 5°C.

R in the compound () used in the present invention is preferably sodium, potassium, barium, calcium or tetraalkylammonium represented by (R ₁ ) ₄ N ⁺ (R ₁ is alkyl having 1 to 4 carbon atoms), particularly sodium , potassium is preferred. The compound () can be produced according to the steps shown below.

The first step is a methylation step. That is, it is a step of converting a solution of the compound represented by formula () into compound () by adding an ether solution of diazomethane or passing diazomethane gas generated outside the system. Examples of the solvent include ether solvents such as ether, 1,2-dimethoxyethane, dioxane, halogen solvents such as THF or chloroform, alcohol solvents such as ethanol and methanol, and ethyl acetate. Among them, ether and ethyl acetate are preferred, but of course they are not limited to these.

The second step is an intramolecular cyclization step. That is, it is a step of reacting a halogenating reagent with a compound of formula () to obtain formula () (in the formula, X represents bromine or iodine). As the halogenating reagent, N-bromosuccinimide, N-iodosuccinimide, potassium-iodine iodide, etc. can be used, but the present invention is not limited to these. Ether solvents such as THF, ether, 1,2-
Dimethoxyethane, dioxane and the like are preferably used, but are not limited to these. The reaction temperature is preferably -10° to 80°C, but is not limited thereto.

The third step is a dehalogenation deuterium step. That is, the formula () (X is the same as the above definition)
This is a step of reacting a compound of formula () with a base to obtain a compound of formula (). The bases used include sodium or potassium acetate, sodium or potassium benzoate, dicyclohexyl-18-crown-6 and sodium or potassium superoxide, sodium or potassium trifluoroacetate, or tertiary amines, among which tertiary amines are used. Preferably, 1,5 etc.
-diazabicyclo[4,3,0]nonene-5,
1,4-diazabicyclo[2,2,2]octane and 1,5-diazabicyclo[5,4,0]undecene-5 are preferably used, but of course they are not limited thereto. Examples of solvents include aromatic hydrocarbons such as benzene, toluene, etc., ether solvents such as ether, THF, dioxane, 1,2-dimethoxyethane, etc.
DMF, DMSO, HMPA, etc. are used, but among them
THF, benzene, DMF, and DMSO are preferred, but are not limited to these. The reaction is generally carried out at a temperature between 10° and 150°, preferably between 25° and 80°C.

The purification is performed by silica gel column chromatography using an organic solvent (preferably ethyl acetate) containing a tertiary amine (preferably triethylamine).

The fourth step is a step of hydrolyzing the methyl ester with a base to obtain the compound () (R is the same as defined above). As the base, sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, barium hydroxide, and calcium hydroxide are used.
Among these, sodium hydroxide and potassium hydroxide are preferred, but of course they are not limited to these.

Compound () can be produced through the above four steps, but in addition to this, a process for directly producing compound () from compound () will be explained in the fifth step. That is,
The fifth step is the formula () (in the formula, X is the same as the above definition)
The compound is dehalogenated with a base, and at the same time, the ester is hydrolyzed to directly form the formula () (where R is an alkali metal, an alkaline earth metal, or (R ₁ ) ₄ N ⁺ (R ₁ is the number of carbon atoms). This is a process for obtaining a compound of tetraalkylammonium (representing 1 to 4 alkyl). Bases include sodium or potassium hydroxide, sodium or potassium carbonate, sodium or potassium bicarbonate,
Alternatively, a superoxide of tetraalkylammonium (R ₁ ) ₄ NO ₂ (R ₁ is the same as above) is preferable, and among these, sodium or potassium hydroxide is particularly preferable, but of course the superoxide is not limited thereto. As the solvent, alcohol solvents such as methanol and ethanol, mixtures of water and ether solvents such as THF, ether, 1,2-dimethoxyethane, and dioxane, DMSO, DMF, and HMPA are used. Particularly preferred are DMSO and DMF, but of course they are not limited to these.

The present invention will be explained in more detail with reference to Examples below, and the following abbreviations are used in the text.

mp: Melting point (uncorrected) IR: Infrared absorption spectrum (only major bands are listed) HMR: Nuclear magnetic resonance spectrum (proton,
100MHz) Mass: Mass spectrum reference example 1 5,6,8,9,11,12,14,15-octadeutero-PGF ₂ methyl ester () 5,6,8,9,11,12,14, 700 mg of 15-octadiutelo-PGF ₂ () was dissolved in 10 ml of ethyl acetate, and a large excess of an ether solution of diazomethane was added under ice cooling, followed by stirring for 10 minutes. The reaction mixture was concentrated, and the resulting oil was purified by column chromatography [silica gel; acetonitrile: ethyl acetate (2:
1)], 680 mg of oil was obtained (yield 97%).

IRνmax (liquid film method): 3350, 1735cm ^-1 Mass (m/e): 358 (M ⁺ ), 340 Reference example 2 5, 6, 8, 9, 11, 12, 14, 15-octadeutero-5 -Iodo-PGF ₁ methyl ester () D-8PGF ₂ 〓() 500 mg was dissolved in 15 ml of chloroform and 1 g of N-iodosuccinimide was added, followed by stirring at room temperature for 1 hour. The precipitate was filtered and thoroughly washed with chloroform, and the liquids were combined, washed with a saturated aqueous solution of sodium bicarbonate, then with a saturated aqueous solution of sodium thiosulfate, and finally with water, dried, and concentrated. When the obtained oil was separated and purified using column chromatography (silica gel; ethyl acetate), 594
mg of D-8-iodoester () was obtained (yield 88.5%).

IRνmax (liquid film method): 3350, 1735cm, ^-1 HMRδ (CDCl ₃ ): 0.90 (3H, t, J = 7.0Hz) 3.67 (3H, S) 5.45 (1H, S) Mass (m/e): 484 (M ⁺ -18) Reference example 3 5,8,9,11,12,14,15-heptadeutero-PGI ₂ methyl ester () 5,6,8,9,11,12,14,15-octa Dieutero-5-iodo PGI ₁ methyl ester ()
Dissolve 590mg in 20ml of anhydrous THF and add 0.99ml of DBU.
Stirred at 50°C under argon for 14 hours. After cooling, filter the precipitate containing 1% triethylamine.
It was washed with THF and the liquid was concentrated. The obtained oil was separated and purified by column chromatography [silica gel treated with triethylamine; ethyl acetate saturated with water containing 1% triethylamine] to give 219
mg of 5,8,9,11,12,14,15-heptadieutero PGI ₂ methyl ester was obtained (yield 49.8
%).

IRνmax (liquid film method): 3350, 1735, 1678 cm ^-1 NMRδ (CDCl ₃ ): 0.90 (3H, t, J = 6.0Hz) 3.68 (3H, S), 5.48 (1H, S) Mass (m/e) :373 (M ⁺ ) Reference example 4 5,8,9,11,12,14,15-heptadieutero-PGI _disodium salt () 5,8,9,11,12,14,15-heptadieutero -Dissolve 60 mg of PGI ₂ methyl ester () in 1.36 ml of methanol containing 1% triethylamine,
0.68 ml of 1N sodium hydroxide was added, and the mixture was stirred at room temperature for 14 hours under argon. This reaction mixture was freeze-dried to obtain 75 mg of the sodium salt of 5,8,9,11,12,14,15-heptadieuteroPGI ₂ (containing 3 times the mole of sodium hydroxide).

IRνmax (KBr): 3350, 1690, 1660, 1560, cm ^-1 NMR (D ₂ O): 0.88 (3H, t, J = 6.0Hz), 5.60 (1H, S) Example 1 5, 8, 9, 11,12,14,15-heptadieutero-6-oxo-PGF ₁ 〓() 5,8,9,11,12,14,15-heptadieuteroPGI _disodium salt () 22 mg (3 times molar (containing sodium hydroxide) was dissolved in 2 ml of water, the pH was adjusted to 3 by adding a saturated aqueous solution of oxalic acid under ice cooling, and the mixture was extracted three times with ethyl acetate. The ethyl acetate layer was washed with water, dried, and concentrated to obtain 16 mg of crystals (mp 60-89°C).

IRνmax (KBr): 3400, 1700, 950cm ^-1 HMRδ (CD ₃ OD): 0.90 (3H, t, J = 5.0Hz), 5.42 (1H, S) Mass (m/e): Hydroxyl group is converted into TMS and ketone M ⁺ 636 of a compound obtained by methoximating and methylating a carboxylic acid.The same compound can be obtained similarly by using hydrochloric acid instead of oxalic acid.

Example 2 5,8,9,11,12,14,15 _- heptadieutero-6-oxo-PGF ₁ 〓() 5,8,9,11,12,14,15-heptadieuteroPGI disodium salt Instead 5, 8, 9, 11,
12,14,15-heptadieuteroPGI _dipotassium salt
Using 20 mg, 13 mg of crystals were obtained in the same manner as in Example 1 (mp 60-89°C).

νmax (KBr): 3400, 1700, 950 cm ^-1 or less The same compound can be obtained by using hydrochloric acid instead of oxalic acid.

Claims (1)

[Claims] 1 General formula () [In the formula (), R represents a tetraalkylammonium substituted with an alkali metal, an alkaline earth metal, or an alkyl having 1 to 4 carbon atoms] Formula () obtained by hydrating a compound represented by 5,8,9,11,12,14,15-heptadieutero-6-oxo-prostaglandin
Production method of F ₁ 〓.