JPS59186986A - Preparation of 1',2'-diacyl-l-biopterin - Google Patents

Abstract

PURPOSE:To prepare the titled compound with decreased amount of solvent and simplified procedure, in high yield, by using 1',1'-dialkylsulfonyl-L-rhamnose as a starting raw material, and carrying out the reaction of the material continuously without separating the unstable intermediate. CONSTITUTION:The objective compound of formula III (R<1> and R<2> are acyl) is prepared by (1) reacting 1',1'-dialkylsulfonyl-L-rhamnose of formula I (R is alkyl) with e.g. NH4OH, (2) reacting the resultant 5-deoxy-L-arabinose with phenylhydrazine to obtain the compound of formula II, (3) acylating the product, and (4) reacting the obtained 2,3,4-triacyl-5-deoxy-L-arabinose-phenyl-hydrazone with 2,4,5-triamino-6-hydroxy-pyrimidine dihydro-chloride. The whole process is carried out continuously without separating the intermediates. USE:Precursor of a drug for the remedy of phenylketonuria, ademonia, Parkinson's disease, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION This invention relates to a method for producing phosphorus. More specifically, L-rhamnose is used as the starting material and the entire reaction process is carried out continuously without isolation of intermediate products 1'.

The present invention relates to a method for producing 2'-diacyl-biopterin.

1',2'-diacyl=L-biopterin is a compound represented by 1 (in the formula, R1 and R2 are acyl groups), and is used for the treatment of L-biopterin and phenylketonuria, depression, Parkinson's disease, etc. It is effective for 1
It is extremely useful as a precursor for -,2'-diacyl-5,6,7,8-tetrahydro-L-biopterin and 5,6,7.8-tetrahydro-L-biopterin.

Examples of the acyl group (R1 and R2), which is a protecting group for the hydroxyl group in the compound represented by formula (Il), include acetyl, propionyl, butyryl, isobutyl, or valeryl group, and those having 1 to 10 carbon atoms are particularly preferred. Generally, R3C0 (wherein R1 is H or a saturated alkyl group containing a linear, branched or cyclic hydrocarbon having 1 or more carbon atoms, preferably 1 to 9 carbon atoms, or R51R
@1R7 and R8 are a hydrogen atom or a straight or branched alkyl group (preferably, the total number of carbon atoms does not exceed 3), a methyl group or an ethyl group, but preferably The total number of carbon atoms does not exceed 2) or a substituted or unsubstituted aralkyl group (which is a methyl group).

R1 and R2 of the compound represented by general formula (1)
is the acetyl group 1-. 2'-Diacetyl-L-biopterin is known (3 ernard schools)
, J ost HB 1eri and
Max Viscontini ; He1
vetica Chimica Acta.

211 (1977)), and other novel diacyl derivatives as well as L-rhamnose, 2,
It can be produced using 4,5-triamino-6-hydroxy-pyrimidine and an acylating agent as main raw materials.

In this case, J, All1er, Chem.

5OC1, Japan-12301 (1975), the hydrate of mono-rhamnose was reacted with ethanethiol to convert 1-rhamnose-diethylmercaptal to L-rhamnose-diethylmercaptal.
．． 1=-Diethylsulfonyl-L-rhamnose is converted to 5-deoxy-arabinose. This was reacted with phenylhydrazine to form 5-deoxy-L-arabinose-phenylhydrazone, and reacted with an acylating agent to form 2.3.4-t-lyacyl-5-deoxy-
Obtain arabinose-phenylhydrazone. This compound is treated with 2.4.5-triamino-6-hydroxy-pyrimidine dihydrochloride and then oxidized with an oxidizing agent such as iodine to obtain 1',2'-diacyl-L-biopterin. L-biopterin can be obtained by deacylating it.

However, when manufacturing according to the above-mentioned known method, a large pot of solvent is required, and the operation is complicated, resulting in a large amount of loss, resulting in a low overall yield, making it unsuitable for industrial processes. It's inappropriate.

The inventors of the present invention
-After rhamnose, the target 1", 2"-diacyl-
A method in which the entire process up to the collection of L-biopterin and, if necessary, L-biopterin is continuously reacted without isolating intermediates, that is, a one pot production method (a method that can be performed in one reactor if necessary) ) was discovered. As a result, the drawbacks of the conventional method were improved, and the overall yield was improved, the amount of solvent used was reduced, and the operation was simplified, resulting in an industrially advantageous production method.

That is, the reaction of the present invention is shown as the reaction formula below, and the present invention converts the starting material 1',1'-dialkylsulfonyl-L-rhamnose (formula (■)) to the intermediate product 5-deoxy -L-arabinose (formula ([1),
5-deoxy-L-arabinose-phenylhydrazone (formula (IV)), 2,3.4-triazyl-5-deoxy-L-arabinose-phenylhydrazone (formula M)
and a compound of formula M and 2,3,4,5-triamino-6
- The reaction proceeds continuously without isolating and purifying the coupling product (Formula 2) with hydroxypyrimidine.

In the reaction of the present invention, the phenylhydrazone derivatives of deoxyarabinose represented by formulas (1) and (M) and the coupling reaction product represented by formula (2) are relatively unstable, so they can be continuously produced by the one pot production method of the present invention. By allowing the reaction to proceed, the yield can be improved.

Further, in the production method of the present invention, it is preferable to oxidize the coupling reaction product represented by formula (2) using a mild oxidizing agent such as iodine, thereby further improving the yield of diacylbiopterin. Furthermore, the formula M
By passing through the triazyl compound represented by , the yield of the coupling reaction product is also improved.

Further, when the production method of the present invention is used, the production of pterin, whose by-product has conventionally been considered a problem, is suppressed, and biopterin can be obtained with a yield that is practically satisfactory.

In this case, the mercaptal used is not limited to ethanethiol, but also methyl mercaptal and propanethiol, and correspondingly 1'.

1'-dimethylsulfonyl-L-rhamnose, 1',
1''-dipropylsulfonyl-L-rhamnose is obtained, which serves as the starting material for the one pot production process.

1",2"-diacyl-L obtained by the method of the present invention
-biopterin, or one obtained by deacylating it; 1', 2--diacyl-5,6,
7,8-tetrahydro-biopterin or 5
,6,7.8-tetrahydro-1-biopterin can be obtained by acylating 1',2-diacyl-L-biopterin or 1-biopterin in a solvent in the presence of a catalyst. Just add hydrogen.

As a catalyst, for example, Yen, NL % Cr SPd
N Rh, etc. may be used.

Hydrogenation is carried out in a solvent, and the solvent used is one in which the compound represented by the ω general formula (11) can be dissolved, such as trifluoroacetic acid, methanol, ethanol, propatool (1), propatool (2), concentrated hydrochloric acid, etc. Even in acidic or basic water, including (b) the compound represented by the general formula m, it does not dissolve, but
, 7,8-tetrahydro-biopterin may be dissolved in, for example, acetic acid.

When using the above-mentioned (ω) solvent, especially trifluorodiacetic acid, see, for example, Bernard, Schulx, Jost
H. Bieri and Max Bisco'/-1-i-ni (
Bernard 3chrs.

Jost HBieri and MaxVis
conNni ; He1vetica Chim
ica Acta.

61(7), 2731 (1978)”)
- It can be carried out according to the method of hydrogenating biopterin under a circular catalyst.

The production method of the present invention is applicable not only to the compound represented by the general formula (I) but also to monabuterin and neopterin having different side chains at the 6-position, and has a wide range of applications.

Next, the present invention will be described in detail with reference to Examples, but the present invention is not limited to these Examples.

Example 1 [1',2''-diacetyl-L-biopterin on
e pot production] (The whole process was carried out under yellow light.) Diethylsulfonyl-rhamnose 14 g (42
, 1 mmol) in 120 ml of water, D I-1
4N-N840H was added with stirring until the value was 9-10. After standing at 22°C for 14 hours with occasional stirring, the diethylsulfonylmethane precipitate was filtered off and the filtrate was dried at 40°C under reduced pressure. Pour the residue into pure methanol 80
5 g of purified phenylhydrazine (46
mmol) was added, left at room temperature for 11 hours, and then heated to 40°C.
and dried under reduced pressure. The residue was washed two to three times with 501 ether per wash, dried, and then dissolved in pyridine 351 and cooled. Then, acetic anhydride 35 was cooled on ice to 0 to 5°C.
1 was added slowly, and after complete addition, the mixture was left in a water bath for about 10 minutes and then left at room temperature for 5 hours. Methanol 2001 was added to this solution -11- and then left at room temperature for 10 to 15 hours (-overnight).

Then sodium dithionite (Na2s, o4) 1,
h JN sodium acetate 3H2o 12.5g water 3
001 and 6-hydroxy-2,4,5-triaminopyridine sulfate H20 in water 5001.
was continuously added to the methanol/pyridine solution. The entire reaction mixture obtained was purged with nitrogen and sealed, and then stirred at 35-40°C for 20 hours to obtain a uniform reddish-brown solution. To this solution, a solution of iodine 25(+) dissolved in 300 ml of methanol was added little by little while stirring to oxidize the formed tetrahydrobiopterin derivative. Excess iodine was removed with sodium thiosulfate (Na2S2O3). After the excess iodine was no longer present, a further small amount of iodine was added to complete the oxidation. During the oxidation process, a brown fine crystal precipitate was obtained. This suspension was concentrated to about 100% After cooling in a refrigerator for several hours, it was filtered. The crude diacetylbiopterin collected by filtration was washed with 50 ml of cold water, 100 ml of cold ethanol, and 100 ml of -11-ether, and dried. The dried residue was poured into about 1200 ml of boiling water. Dissolves and activates secondary 5
It was decolorized with g. This solution was filtered while heated, the activated carbon portion was washed with 501 ml of boiling water, and the filtrate was allowed to cool to room temperature and then kept at 0 to 5°C for 10 hours.

The precipitated diacetyl-L-biopterin crystals were collected by filtration,
Washing twice with 501 g of ethanol and then with ether and drying yielded 8.10 g of crystals of 1',2-diacetyl-biopterin. The yield was 60%.

Example 2 [1-,2-dibutyryl-L-biopterin on
e pot production] (The whole process was carried out under yellow light.) Diethylsulfonyl-L-rhamnose 14 (1 (42
, 1+n+nol) was suspended in water 1201, and Ell-
4N-NH40H was added with stirring until 1 was 9-10. After standing at 22°C for 14 hours with occasional stirring, the diethylsulfonylmethane precipitate was filtered off.
The filtrate was dried at 40°C under reduced pressure. The residue was dissolved in pure methanol 801 and purified phenylhydrazine 5 a
(46 mmol) was added thereto, allowed to stand at room temperature for 1 hour, and then dried under reduced pressure at 40°C.

The residue was washed 2-3 times with so ml of ether each time, dried, then dissolved in pyridine 351 and cooled. Then, 35 ml of ice-cooled butyric anhydride was slowly added to 0 to 5°C, and after complete addition, the mixture was left in a water bath for about 10 minutes, and then left at room temperature for 5 hours. 200+nl of methanol was added to this solution, and the mixture was further left at room temperature for 10 to 15 hours (-overnight).

Next, dithionite, silium, Og and sodium acetate, 3 H2012,5g, water 300ml, 2WI
Add 6-hydroxy-2,4 to 500 ml of water and the dissolved solution.
, 5-triaminopyridine sulfate)+20 was continuously added to the methanol/pyridine solution. The entire reaction mixture obtained was purged with nitrogen and sealed.
Stirring at 35-40°C for 20 hours yielded a homogeneous reddish-brown solution. A solution of iodine 25Q dissolved in methanol 3001 was added little by little to this solution while stirring, to oxidize the produced tetrahydrobiopterin derivative.

Excess iodine was removed with sodium thiosulfate, and once no excess iodine was present, a further small amount of iodine was added to complete the oxidation. A brown fine crystal precipitate was obtained during the oxidation process. This suspension was heated to 501 °C under reduced pressure at 40 °C.
After concentrating and filtering, the filtered insoluble fraction was washed with water, cold ethanol 401 and ether, and recrystallized from hot ethanol. Activated carbon was used for this WA sharpening. That is, a mixture of activated carbon and crystals is heated and filtered, the filtrate is cooled, a yellow precipitate is collected by filtration, washed with ether and dried to obtain 1'.

10.3 g of 2'-dibutyryl-L-biopterin was obtained. The yield was 65%.

Next, the characteristic values of the obtained compound are shown. Also I H-N
Figure 1 shows the details of the MR spectrum analysis.

Ry value (3%N84C1/VbO, cellulose)
−0,36= 14− [α]22o −−74,3±3° (C−0,8,
IN-8 HC1) (The optical rotation increased with time for the ice thaw of the butyrate ester.) 1) -1-NMR spectrum analysis (δ value: ppI!
1): (90MH2, INDCj) 9.33 (s, H-Cf force), 6.5 (d,
) I-C (1M), 5-6 (IIl, N20, H-
C(2')), 3-2.66 (III, 2
H2C-(CH2-DH5)), 2.30-1.8(
11゜2H2C(CH3)), 1.66 (d,
N3-C (3M), 1.33 (j, N3C-
(C)12)) Elemental analysis value: C engineering. N23 N505 Theoretical value (%): 0 54,11 H6,I N18
．． 56 Actual value (%): C54,0986,99N18.99 Example 3 [One pot production of L-biopterin] (All steps were performed under yellow light.) 15-Diethylsulfonyl-L-rhamnose 14 ( 1 (42
, 1 ml) was suspended in water 1201, and the DH was 9 to 1.
4N-NHa OH was added with stirring until the temperature reached 0. After being left at 22°C for 14 hours with occasional stirring,
The precipitate of diethylsulfonylmethane was filtered off, and the filtrate was
It was dried under reduced pressure at 0°C. The residue was dissolved in methanol 801 and purified phenylhydrazine 5 g (46
mmof) and left at room temperature for 1 hour, then heated to 40°C.
and dried under reduced pressure.

The residue was washed 2-3 times with 501 ether each time and dried, then dissolved in pyridine 351 and cooled. Then, ice-cooled acetic anhydride 351 at 0-5°C was slowly added,
After finishing, it was left in a water bath for about 10 minutes and then left at room temperature for 5 hours. Methanol 2001 was added to this solution, and the mixture was further left at room temperature for 10 to 15 hours (-overnight).

Next, a solution of 1.0 g of sodium dithionite and 382012.5 g of sodium acetate dissolved in 3001 parts of water and 6-hydroxy-2,4,5-triaminopyridine sulfate N20 in 500 parts of water were suspended to make a saliva solution. Continuously added to the methanol/pyridine solution. The entire reaction mixture obtained was purged with nitrogen, sealed, and stirred at 35-40°C for 20 hours to obtain a homogeneous reddish-brown solution. A solution prepared by dissolving 25 g of iodine in methanol 3001 was added little by little to this solution with stirring to oxidize the tetrahydrobiopterin derivative being produced.

Excess iodine was removed with sodium thiosulfate, and once no excess iodine was present, a further small amount of iodine was added to complete the oxidation. A brown fine crystal precipitate was obtained during the oxidation process. This suspension was concentrated to about 100 ml and mixed with methanol 150 1111 and 14N-NHt OH25
0w+l was added and kept at 50°C for 1 hour to deacetylate. The solution was evaporated to dryness under reduced pressure at 40°C, collected in 1001 methanol and filtered. The obtained crude biopterin was washed with ice-cold water 501 and ethanol 2001, and dissolved without drying in 1400 ml (minimum volume) of boiling water to which activated carbon had been added. This solution was filtered while still hot, and the filtrate was allowed to cool to room temperature and then left at 5° C. for 10 hours. The biopterin crystals obtained were filtered off, washed with cold water, ethanol and ether, and placed under reduced pressure (0.01 Torr).
The mixture was dried at 40° C. for 14 hours to obtain 6 g of L-biopterin. The yield was 60%.

Reference Example 1 [Production of 1',2-diacetyl-5,6,7,8-tetrahydro[1-L-biopterin] P was prepared using hydrogen in trifluoroacetic acid 501 by a conventional method.
After reducing 350 mg of t o, 10 of pure 1',2''-diacetyl L-biopterin was added to the suspension. Hydrogen was then passed through and after 40 minutes the hydrogen blowing rate was reduced and after 45 minutes The circles were immediately filtered off and the colorless filtrate was frozen in liquid nitrogen. A cold mixture of methanol 20III and 3 ml of 12N-HC# was then added to the frozen solution under a stream of nitrogen or argon gas, and further ether was added. 1801 was added. When the frozen solution was thawed at room temperature, 1',2'-diacetyl-(6R1S)-5,6,7,8-tetrahydro-
L-biop16-thelin.2HC# was isolated as a white powder.

The powder was centrifuged, washed with acetonitrile and ether, dried in a desiccator using KOI-1, and then dried at 22° C. under reduced pressure (0.01 Torr) for 15 minutes.
Dry for an hour (yield: 1.11 J>).

The obtained 1-,2-diacetyl-(6R,5)-5,
IH-6,7,8-tetrahydro-L-biopterin
NMR spectroscopy (ds-pyridine) and
Charts obtained by C-NMR spectrum analysis (D20) are shown in FIGS. 2 and 3, respectively.

As is clear from Figures 2 and 3, the obtained compound is 1'
, 2''-diacetyl=(6R) -5,6°7.
8-tetrahydro-L-biopterin and 1', 2'-
It was an approximately 1:1 mixture of diacetyl-(+38)-5,6,7,8-tetrahydro-L-biopterin.

Reference example 2 [1-,2” ~ dibutyrolu (6R,S) -5
, 6゜7.8-Tetrahydro-L-biopterin production] 19- 250 mg of PtO was stirred in 25 ml of purified acetic acid at room temperature in a hydrogen stream for about 10 minutes to completely saturate the PtO and catalyst with hydrogen, and then 1-. 2'-dibutyl-
500 mg of L-biopterin was added. Dibutyryl-L
- Biopterin does not dissolve in acetic acid and becomes a suspension; however, by hydrogenation with stirring, a tetrahydro derivative is generated and the solution becomes a solution, and after 5 hours it becomes a clear solution. The catalyst was filtered out from the resulting solution, and the solution was frozen using salt and ice as a cooling agent. After freezing, 12N-H (
A solvent consisting of Jlml, methanol 91, and ether 3001 was added, and the temperature of the entire flask was returned to room temperature. The precipitated dibutyryl-tetrahydro-L-biopterin dihydrochloride was collected by filtration, washed with ethanol and then ether until excess HCρ was removed, and washed under reduced pressure (0,
The solvent was removed by drying at 60°C for 16 hours. The yield was almost quantitative for dibutyryl-L-biopterin.

The obtained 1',2'-dibutyryl-(6R,S)-54
,7,8-tetrahydro-biopterin 11
Char 1 of the 11-1-N spectrum analysis (in INDCf) is shown in FIG.

As is clear from FIG. 4, the obtained compound is 1-. 2
'-dibutyryl-(6R) -5,6,7,8-tetrahydro-biopterin and 1=,2=-dibutyryl-(68) -5,6,7,8-tetrahydro-
It was an approximately 1:1 mixture of biopterin and biopterin.

Reference Example 3 [Production of (6R,S')-5,6,7,8-tetrahydro-shi-biopterin] 10 mg of Pt02 was stirred in purified acetic acid 301 for about 10 minutes at room temperature in a hydrogen stream to prepare a Pt02 catalyst. After being completely saturated with hydrogen, purified L-biopterin 500I
11 g was added. Although L-biopterin was not dissolved in acetic acid and a suspension was obtained, this was stirred in a hydrogen stream until it became a solution. It became a solution in about 5 hours. The catalyst was filtered off from the resulting solution, and the solution m was cooled on ice using a cryogen until solid i was formed. After solids were formed, methanol 91, ether 901 and 12N-HC#11
The following solvent was added and the temperature of the entire flask was returned to room temperature. The precipitated dihydrochloride of tetrahydrobiopterin was collected by filtration, and the excess liter was dissolved using ethanol and then ether.
-I Washed until free of CI and dried under reduced pressure.

The crude tetrahydrobiopterin obtained was crystallized from acetic acid and ether and then heated under reduced pressure (0.01T) to remove the solvent.
orr) at 60° C. for 16 hours. The yield was almost quantitative for biopterin.

[Brief explanation of the drawing]

FIG. 1 is a chart of the IH-NMR spectrum of 1',2'-dibutyryl-biopterin obtained in Example 2, and FIGS.
2”-Diacetyl-(6R,S)-5,6,7
,8-tetrahydro ~I H- of L-biopterin
NMR spectrum analysis and 13C-NMR spectrum analysis chart, Figure 4 shows 1- and 2 obtained in Reference Example 2.
--Dibutyryl-(6R1S)-5,6,7, is a chart of IH-NMR spectrum analysis of tetrahydro L-biopterin. -n J-

Claims (1)

[Claims] 1 1', 1-Dialkylsulfonyl-rhamnose is converted to 5-deoxy-L-arabinose, then treated with phenylhydrazine to form 5-deoxy-arabinose-phenylhydrazone, and this is further acylated. , produced 2,3.4-t-lyacyl-5
-deoxy-arabinose-phenylhydrazone was reacted with 2.4.5-1-riamino-6-bitroxy-pyrimidine dihydrochloride and then oxidized to give 1-. A method for producing 1',2-diacyl-L-biopterin, characterized in that all reaction steps are carried out continuously without isolating intermediates. The manufacturing method described in paragraph 1.