Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
- 1 - 1 338 1 4~
This is a divisional application of copending
application 584, 504, filed November 29, 1988.
The Present invention relates to intermediates f~r sYn-
thesizin~ 8H4 and its d~rivatives, as well as to a Process for
the PreParation of L-bioPterin.
The comPound 8H4, namelY, 5,6,7,8-tetrahYdro-L-erYthro-
bioPterin, is one of the coenzY~es of aromatic amino acid
hYdroxYlase, is an indisPensable comPound for ~iosYnthesis of
doPamine, noradrenalin, adrenaline, and melatonin, and is
rePrsented bY the followins formula:
~H H H OH
~ N ~ CH3
H2NJ~n~NJ dH
lS H
A deficiencY of BH4 causes serious neurolosical disorders
li~e atYPical PhenYlketonuria and Parkinsonism: and recentlY, it
has been found that sYmPtO~s due to such diseases can be
renarkablY imProved bY administration of BH4. Further, it has
been recosnized that BH4 is effective for curing infantile
autism and dePressions.
Since BH4 has such useful Pharmacolosical activities,
various studies have been made for the sYnthesis thereof, and
are referenced as follows: E.L.Patterson et al. "J. Am. Che~.
Soc." Vol.78, page S868 ~1956); H. Re~bold et al. "Chem. Ber.
Vol.96, Paoe 1395 (1963); E.C. TaYlor et al. "J. A~. Chem. Soc.
Vol.98, Page 2301 (1976~: M. Viscontini et al. ~Helv. Chi~.
Acta" Vol.52, Page 1225 (1969); ibid., Vol.55, Pase S74 ~1972);
ihid., Vol.60, Page 211 (1977); ibid., Vol 62, Pase 2S77 ~1979);
K.J.M. Andrews Q~ al. ~J. Chem. Soc.n (c), Pa~ 928 11969); S.
Matsuura et ~1. ~8ull. Che~. Soc. JPn.n~ol.48, Pa~e 3767 (1975);
ibid., Vol.52, Pa~e 181 (1979) ~Che~. Lett.~ pa~e 735 (1984);
Jap. Pat. No. 59 - 21685(A); ibid., 59 - 82091(A); and ibid., 60
1 3381 49
- 204786(A).
Each of the conventiona~ Processes for PreParin~ BH4 has
several trawbacks in that an expensive saccharide is required as
startins material to Provide the asYmmetric carbon atom at its
side-chain, that Yield and PuritY will become low due to multi-
reaction stePs, throush unstable intermediates that require
troublesome treatment oPerations, and that it requires
troublesome Purification Procedures. Therefore, the conventional
Processes are unsuitable for industrial Production of the co~-
pound and its derivatives.
The Present inventors have enerseticallY worked to develoP a
Process for PreParins 8H4, one which can be aPPlied for a co~-
mercial and convenient Production thereof, bY takins the follow-
in~ Points into consideration:
a) The startins material is available at reasonable cost,
b) The number of sYnthetic stePs can be made minimum,
c) Good Yield can be attained, and
d) If Possible, it can be applied for sYnthesizins comPounds
analosous to BH4.
As a result, theY have found that a tetrahYdrofuranYI-
PYrimidine derivative of the formula
ORl H
NzN/~Xr' ;~oJ~0113
H2 R4
wherein R1 is a hYdrogen atom, alkYI, aralkyl, or arYI 8rouP;
R2 is an alkyl, hYdroxYalkYI, or PolYhydroxyalkyl srouP; R3
and R4 are the same or different and rePresent alkyl, aralkYI
or arYI ~rouP.
is suitable as an intermediate for sYnthesizins BH4 and
anal 090US comPounds thereof to oPen the waY for the Present
invention~
1 3381 ~9
The following are steps for synthesizing the
tetrahydrofuranylpyrimidine derivatives shown by said Formula
I as well as BH4 and its analogous compounds:
OR1
~ ~H2SO4 + ~ ~ R2
H2N N NH2 R40 0 OR3
(Vlll) - (IX)
~,
ORl H
H2N ~ ~ /N ~ OR3
H2 R4
(I)
t 33 ~ 1 ~9
(I)
ORl H OH ORI H OH
H N/l\~NX ~R2 _ ~XY ~ R2
(111') \ (IV')
v ~
ORl R6 OR5 / ORl OH
nHX ~/ ~/R2
R7HNJ~N~( N /2~lJ~NJ\N~J O nHX
ORl ~R6 OR5 ORl OH
R~HN/~XN~ H NJ\~(N~Rz
R8
(IV") (Vl)
ORl H OH
N~( ~ R2
(Vl I )
OH OH OH OH
H N1~XNJ~ H2N1~XN~/~
(V'-a) (Biopteri ne)
1 3381 49
wherein R1, R2, R3, and R4 have the meaninss as already
stated; Rs, R6, R7, and R~ are the same or different and
rePresent a hYdrosen atom or acYI srouP: Rg is an alkYI,
aralkyl or arYl ~rouP; n is an inte~er of 5 or less; and HX
/ 5 is an acid.
In connection with the comPounds, the definitions of each
substituent shall ~enerallY be siven as follows: The alkYI srouP
maY be of methYI, ethYI, ProPyl, butYI, PentYI, hexYI srouP,
isomers thereof or the like. The aralkYI ~rouP maY be of benzYI,
xYlYI, PhenethYI srouP or the like. The arYI srouP maY be of
PhenYI, tolYI, anisoyl, naPhthYI sroup or the like. The
hYroxyalkYI srouP maY be of hYdroxYmethYI, hYdroxYethYI,
hydroxypropyl srouP or the like. The polYhYdroxYalkYl srouP maY
be of 1,2-dihYdroxYethY1, 1,2-dihYdroxYProPyl~ 2,3-dihYdroxY-
propyl, 1,2,3-trihYdroxyproPYI sroup or the like.
The tetrahydrofuranylpyrimidine derivative (I) can be
prepared by reactins 2,4,5-triaminoPYrimidine derivative (Vlll)
with an ePoxY-tetrahYdrofuran derivative (IX).
The tetrahYdrofuranYIPYrimidine derivative (I) can be con-
verted into dihYdroPteridine derivative (Ill') or tetrahydro-
pteridine derivatiYe (IV') bY hYdrolYzin~ its acetal Portion
with an acid, and bY rins closing at the amino ~rouP Portion.
The dihYdroPteridine derivative (Ill') can be converted into
.. . . . . . . .. . . . . . . .
pteridine derivative (V') bY oxidizing its Pteridine rins, or
into acyl-dihYdropteridine derivative (Ill'') throu~h an acYla-
tion. The acyl-dihYdroPteridine derivative (Ill'') can easily be
changed bY treatin~ with an alcohol into acYI-tetrahYdro-
pteridine derivative (IVn) which can, in turn, be converted
again into the dihYdroPteridine derivative (Ill') bY treatins
the same with an acid. While, the tetrahYdroPteridine derivative
(IV') can be transformed into dihYdroPteridine derivative (Ill')
bY treatins the same with an acid, or into acYltetrahYdro-
pteridine derivative (IV") through an acYlation thereof.
The Pteridine derivative (V') can be converted into bio-
Pterin derivative (Vl) bY reducin~ the carbonYI srouP in its
side-chain. This reductian can be carried out with the use of
- 6 - l 3 3 8 1 ~ 9
sepiapterin reductase in the manner as described by S. Katoh
et al. "Biochem. Biophys. Res. Commun." Vol. 118, page 859
(1984), or with use of a conventional reducing reagent. In
this connection, a conversion from the biopterin derivative
(VI) to tetrahydrobiopterin derivative (VII) has already been
reported in various papers, for instance, S. Matsuure et al.
"Chem. Lett." page 735 (1984), and Japanese Pat. Nos. 59 -
21685 (A), 59 - 82091 (A) and 60 - 204786 (A), dated Feb,
1984, May 1984 and Oct. 1985, respectively.
According to the invention, L-biopterin can be prepared
by treating biopterin derivative (V'-a), wherein R1 tof (v')]
is a hydrogen atom, and R2 is a methyl group, with
sepiapterin redutase (Enzyme Code No. EC 1.1.1.153). In
connection with this, please note that the reduction of 2-
amino-4-hydroxy-6-(1'-hydroxy-2'-oxopropyl)pteridine (V'-a)
shall, in general, produce four compounds; but according to
the invention, L-biopterin (compound 1) can only and
unexpectedly be formed with a stereospecificity.
_ 7 _ 1 3381 ~9
OH OH
~ N ~ Me
OH OH /~zNJ~NJ~\N~J OH
N ~ ~ Me (1)
I l O OH OH
H2N ~ N ~ \N ~ ~
HzN N N
(2)
OH OH
16 OH OH ~ ll2N~ Me
N ~ Me (3)
O OH OH
HzN " ~N~N~Y ~ N ~ N ~ Me
H2NJ~NJI\N~J OH
(4)
.. . .
In the above sYnthetic Procedures, BH4 can be PrePared as
the final product (Vll), when Rl is a hYdrogen atom and R2 is a
- methyl grouP. NeoPterin is also sYnthesized by this Prodedure,
when Rl is a hYdrogen atom and R2 is a hydroxYmethYI grouP.
Therefore, BH4 and its analogous comPounds can be sYnthesized
easily and at a reasonable cost, when the comPounds according to
the invention are utilized as an intermediate therefor.
- 8 - 1 3381 49
The invention will now be further exPlained in more detail
with reference to Examples.
Exam D le 1
a) 2,5-DihYdro-?,5-direthoxY-?-methYIfuran
To a mixture of 2-methYlfuran (1239, 1.50mol), sodium car-
bonate (318s, 3.00mol) and methanol (2180ml) cooled at -75~
under stirring in a dry ice~acetone bath, dichloromethane
(87.9ml) containin~ bromine (240~, 1.50mol) was added droPwise
over 4 hours.
After comPletion of the addition, the reaction mixture was
removed from the bath and allowed to come to room temPeratUre;
it was then filtered to remove sodium bromide. Saturated sodium
chloride solution was added to the filtrate, wich was extracted
with dichloromethane 3 times. The combined dichloromethane laYer
was washed twice with saturated sodium chloride solution and
dried over anhYdrous sodium sulfate. The resultins solution was
concentrated in a water bath kePt at S0-C or less and distilled
in vacuo to afford 171~ (79.1%) of the titled comPound.
Boilins Point : 54C (bP14)
lH-NMR sPectrum (CDC13, ~ PPm) :
1.51, 1.57 (3H, s x 2, -CH3)
3.12, 3.20 (3H, s x 2, -OCH3)
3.43, 3.50 (3H, s x 2, -OCH3)
5.48, 5.76 (lH, Proton at 5-Position)
5.96 (2H, m, olefinic proton)
Silica sel TLC :
Rf = 0.53,0.59 (hexane : ethYI acetate = 5 : 1)
b) 3,4-EPoxY-?,5-dimethoxY-?-methvltetrahYdrofuran
To a mixture of 2,5-dihYdro-2,5-dimethoxY-2-methYIfuran
(21.69, 150mmol), benzonitrile (17.09, 165mmol), and methanol
(40ml) heated at 40-C , 31% aqueous solution of hYdrosen Peroxide
(11.0g, 100mmol) was added droPwise over 2.5 hours, while main-
taining PH in a ranse of 7.50 to 8.00 bY addition of an aqueous
solution of sodium hYdroxide. Then the mixture was further
reacted for 17.5 hours, while maintainin~ the temPerature at 60
C and pH in the ran~e of 7.50 to 8.00.
1 3 3 8 1 49
After confirmation of the exhaustion of the Peroxide bY
iodometrY, the reaction mixture was cooled to room temperature,
60ml of water was added to it, and it was extracted three times
with chlorofor~. After the extract was washed with water three
times, pentane was added to the chloroform layer to remove
benzamide. The resultin~ filtrate was concentrated to obtain a
crude Product (30.79), which was subiected to silica sel
chromatosraPhY (elution solvent, hexane : ethYI acetate = 6 :
1), so as to afford 2.219 (9.2X) of the titled comPound.
101H-NMR sPectrum (CDC13 ~ PPm) :
1.51 (3H, s, -CH3)
3.33 (3H, s, -OCH3)
3.47 (lH, d, J = 2.7Hz, Proton at 3-Position)
3.5~ (3H, s, -OC~3)
3.67 (lH, br.d, J = 2.7Hz, Proton at 4-Position)
5.10 (lH, br.s, Proton at 5-Position)
Silica sel TLC :
Rf = 0.~7 (hexane : ethYI acetate = 2 : 1)
c) ?,4-Diamino-6-hydroxv-5-[4'-(3'-hydroxy-2',5'-djmethoxy-
20~'-methYltetrahydrofuranyl)]pyri~idine
To a susPension of 2,4,5-triamino-6-hYdroxYPYrimidine sufate
(8.669, 36.2mmol) in ethanol (18ml), sodium carbonate (7.69~,
72.5mmol) dissolved in distilled water (72.5ml) was added. After
.. . . . .
completion of bubblins with carbon dioxide sas, 3,4-ePoxY-2,5-
25dimethoxY-2-methyltetrahydrofuran (2.909, 18.1 mmol) dissolved
- in ethanol (18.3ml) was added, and the resulting mixture was
heated to 100~ under nitrogen atmosPhere to react for 69 hours.
After the mixture was cooled to room temPerature, methanol
(50~1) was added to it and solids were removed bY filtration.
30After concentration of the filtrate, methanol (50ml) was added
and then insoluble mattcr was filtered off. The resultins
fiItrate was concentrated to obtain a crude Product which was,
in turn, subiected to latrobeads column chromatograPhY (elution
solvent, methanol) to afford 2.949 (53.9~) of the titled com-
pound.
lH-NMR sPectrum (PYridine-Ds, ~ PPm) :
-- 10 --
1 338 1 4 9
1.74 (3H, s, -CH3)
3.-4~ (3H, s, -OCH3)
3.49 (3H, s, -OCH3)
3.81 ~1H, m, proton at 4'-Position)
4.71 (1H, br.d, J = 6Hz, Proton at 3'-position)
5.38 ~1H, br.d, J = 5Hz, Proton at 5'-position):
3C-NMR sPectrum ~D~S0-D~, ~ pPm) :
19.4, 48.1, 5S.2, 70.1, 77.2, 98.1, 107.0, 108.0, 152.2
160.9, 161.4
IR sPectrum (KBr, cm-1~ :
., ,
3328, 1588
MS sPectrum (El) :
301(MI), 269
Silica sel TLC :
Rf = 0.27 (chloroform : methanol - 5 : 1)
E~m~le
~-Amino-4-hYdroxY-7-methoxY-6-(1'-hYdroxy-7~-oxopropyl)
5,6,7,8-tetrahvdroPteridine
To 2,4-diamino-6-hYdroxY-5-~4'-(3'-hYdroxY-2~s~-dimeth
2'-methYltetrahYdrofuranyl)~pyrimidine (70ms,0.232mmol) obtained
bY the Process describet in Example 1, acetic acid (3.0ml) was
added, and the mixture was stirred at 50~ for 20 ~inutes. After
beins cooled to room temPerature, the reaction mixture was dried
in vacuo to obtain a crude Product which was, in turn, subiected
to silica sel thin-laYer chromato~raPhY (develoPins solvent,
chloroform : methanol= 5 : 1) to afford 17.6m~ (28.2X) of the
titled compound.
- 11 - 1 338 1 49
H-NMR spectru~ (DMSO-06, ~ PPm):
2.22 (3H, s, -CH3)
3.03 (lH, ~, proton at 6-position)
3.49 (3H, s, -OCH3)
4.13 (lH, br.t, iroton at 1'-Position)
4.88 ~lH, d, J = 6Hz, Proton at 7-Position)
.69 (lH,br.d, J = 6Hz, hYdr`oxY Proton at l--Position)
3C-NMR sPeCtrUm (D~SO-D6, ~ ppm) :
26.6, 55.6, 55.7, 74.4, 96-.9, 98.5, 155.9, 157.7, 158.1
1~ 210 4
MS sPeCtrU~ (El) :
269(M+~, 196
Silica sel TLC :
Rf = 0.48 (chlorofo-~ : methanol = 5 : 1)
E~mPle 3
?-Acetamjdo-5-acetyl-4-hvdroxy-7-olethoxy-6-(1-hvdroxY-?'-
oxoProPvl)-5,6,7,8-tetrahYdroDteridine
To 2,4-diamino-6-hYdroxY-5-t4'-(3'-hYdroxY-2~5--dimethoxy-
2'-methYltetrahydrofuranYl)]pyrimidine (220ms, 0.730mmol) ob-
tained bY the Process described in Example 1, 6N hYdrochloricacid (2.20ml) was added, and the mixture was stirred for 10
minutes at room temPerature and then concentrated in vacuo
After the concentrate was cooled on ice, acetic anhYdride
(3.50ml) and PYridine (3.50ml) were added and allowed to react
for 1 hour at room temPerature under stirrins. After removal of
insoluble matter bY filtration, the filtrate was Poured into
ether (60ml), and the semi-solid material that aPPeared was col-
lected bY decantation. The material was then washed three times
with ether and dried n vacuo. To the dried 0aterial, msthanol
3~ (7ml~ was added and allowed to react for 30 minutes at room tem-
Perature under stirrin~. The reaction mixture was subiected to
silica ~el thin-laYer chromatosraPhy (develoPing solvent,
chloroform : methanol = ~ to a~ford 73.9ms (28.8%) of the
titled comPound.
H-NMR sPectrum (D~SO-06, ~ PPm) :
1.96, 1.99, 2.14 (3H x 3, s x 3, CH3CO x 2, -CH3)
- 12 - ~ 3381 49
3.19 (3H, s, -OCH3)
4.-58 (lH, br.d, J = 4.5Hz, proton at 7-
Position)
4.86 (lH, d, J = 7.3Hz, Proton at 1'-
position)
5.22 (lH, br.d, J = 7.3Hz, Proton at 6-
position)
7.91 tlH, br.d, J = 4.5Hz, Proton at 8-
Position)
13C-NMR sPectrum (DMSO-D6, ~ ppm) :
20.2, 21.7, 26.5, 47.5, 53.5, 74.7, 81.9. 92.7, 153.0
153.3, 157.4, 169.5, 171.0, 204.1
IR sPectru~ (K8r, cm-l) :
3344, 1710, 1616
Silica sel TLC :
Rf = 0.27 (chloroform : methanol = 5 : 1)
ExamPle 4
~-Acetamido-4-hvdroxv-7-methoxY-6-(1'-acetoxY-2'-oxoProPvl)-
5,6,7,8-tetrahvdrooteridine
To 2,4-diamino-6-hYdroxY-5-t4'-(3'-hYdroxY-2~5~-dimeth
2'-methyltetrahvdrofuranyl)]pyrimidine (150m~, 0.498mmol)
obtained by the Process described in Example 1, acetic acid
(1.5ml) was added, and the mixture was stirred for 20 minutes
at SO~ . After bein~ cooled to roo~ temPerature, the reaction
mixture was dried in vacuo. A Part of the dried substance was
taken and subiected to 1H-NMR spectroscoPy to confirm the forma-
tion of 2-amino-4-hYdroxy-7-methoxy-6-(1'-hydroxY-2'-oxoProPyt)
5,6,7,8-tetrahYdroPteridine (see ExamPle 2).
After the other Part was ice-cooled, acetic anhYdride
(2.5ml) and PYridine (2.5ml) were added, and the mixture was
stirred for 1 hour at room temPerature; then ether (SOml) was
added. The resulting solid material was collected bY filtration
and washed with ether to obtain 94.1m~ of a crude product which
was subiected to silica gel thin-layer chromatograPhY
(developins solvent, chloroform : methanol = S : 1) to afford
20.0ms (11.4%) of the titled comPound.
- 13 -
1 33~1 49
lH_NMR sPectru~ (D~S0-~6~ ~ PP~)
2.~1, 2.10, 2.17 (3H x 3, s x 3, CH3CO x 2, -CH3)
3.42 (3H, s, -OCH3)
3.83 (1H, m, Proton at 6-Position)
4.~6 ~1H, br.d, J = 6.8Hz, Proton at 5-
pos i t i on)
5.lB ~1H, d, J = 2.0Hz, proton at 7-
Position)
5.28 ~1H, d, J = 3.4Hz, Proton at 1'-
, Position)
3C-NMR sPectrum ~D~SO-D6, ~ PPm) :
20.3, 24.1, 27.0, 52.4, 55.3, 77.7, 96.6. 103.6, 148.3
152.4, 155.0, 16~.9, 169.6, 203.9
MS sPectrum ~EI) :
lS 353~MI)
Silica gel TLC :
Rf = 0.45 ~chloroform : methanol = 5 : 1)
~xa-P I e 5
2-Amino-4-hvdroxY-6-(1 '-hYdroxY-~'-oxoProPyl)-5~6-dihydr
Pteridine
To 2,4-diamino-6-hYdroxY-5-14'-~3'-hYdroxY-2~5~-dimethoxy-
2'-methYltetrahYtrofuranYI)]Pyrimldine ~lOms, 0.033mmol) ob-
tained bY the Process described in ExamPle 1, 6N hYdrochloric
acid (100~ I) was added, and the mixture was stirred for 10
minutes at room te~perature and immediatelY concentrated in
vacuo to afford 9ms of the titled comPound as crude Product.
This substance exsits in the followins chemical ecuilibrium
in hYdrochloric acid solution:
- 14 - 1 3381 ~9
OH H OH OH H OH
H2N ~ ~ N ~ ~2~ 1 ~ N
H-NMR sPectrum (~OX DCI, ~ PPm) :
2.51, 2.54 (3H, s x 2, -CH3)
4.17, 4.37 (lH, br.d, br.s, J = 7Hz, proton at 6-
position)
4.88 (br.d, J = 7Hz, proton at 1'-Position)
5.65, 5.74 (lH, br.s x 2, Proton at 7-position)
IR sPectrum (KBr, cm-1) :
3260, 1650
MS sPectrum (FAB, Positive)
238[(M+H)+], 165
ExamPle 6
2 ~ ino-4-hYdroxY-6-(1'-hYdroxY-2'-oxoProPyl)-5~6-dihydr
Pte ridine
To 2-amino-4-hYdroxY-7-methoxY-6-(1'-hYdroxy-2~-oxopropyl)
5,6,7,8-tetrahYdropteridine (30ms, 0.1Ommol) obtained bY the
Process described in Example 2, 6N hYdrochloric acid (600~ I)
was added, and the mixture was stirred for 10 minutes at room
temperature and concentrated i~ vacuo to afford 23ms of the
titled comPound as crude Product.
Physico-chemical data of this comPound were the same as
those disclosed in Example 5.
ExarP I e 7
7-Aceta~ido-5-acetvl-4-hydroxY-6-(1'-hYdrOXY-~'-OXOPrODYI)-
5,6-dihvdroPteridine
To 2,4-diamino-6-hYdroxY-5-t4'-(3'-hYdroxY-2~5~-dimethoxY-
2'-0ethYltetrahydrofuranyl)]PYrimidine (220ms, 0.730mmol) ob-
tained bY the Process described in Example 1, 6N hydrochloric
acid (2.20ml) was added, and the mixture was stirred for 10
minutes at room temPerature and then dried in vacuo. After the
material was cooled on ice, acetic anhYdride (3.50ml) and
- 15 ~ 1338t49
pyridine (3.50ml) were added to the concentrate and allowed to
react fo-r 1 hour under stirrins. After insoluble matter was
filtered off, the filtrate was Poured into ether (60ml), and the
resulting semi-solid material was obtained bY decantation, which
material was washed three times with ether and dried in~va~ to
afford 400m~ of the titled compound as crude Product.
H-NMR sPectrum ~PYridine-Ds, ~ PPm) :
2.15, 2.34, 2.54 (3H x 3, s x 3, CH3CO x 2, -CH3)
5.62 (lH, d, J = 8Hz, Proton at 1'-
Position)
5.96 (lH, br.s, Proton at 7-Position)
6.24 (lH, br.d, J = 8Hz, Proton at 6-
Position)
MS sPectrum (FAB, Positive) :
322 [(M~H)+]
~mPle 8
?-A-ino-4-hvdroxY-6-(l'-hYdroxY-~-oxoDropyl)-5~6-dihydr
Pteridine
To 2-acetamido-5-acetYI-4-hYdroxy-7-methoxY-6-(l'-hydroxy-
2'-oxoPropyl)-5,6,7,8-tetrahYdropteridine (lOms, 0.028mmol)
obtained by the process described in Example 3, 6N hYdrochloric
acid (300~ 1) was added, and the mixture was stirred for 6 hours
at room temperature and then concentrated in vacuo to afford
. . .
6.5m~ of the titled comPound as crude Product.
This comPound gave physico-chemical data the same as those
- listed in ExamPle 6.
Exa-Ple 9
~mino-4-hvdroxv-6-(l~-hvdroxy-?~-oxopropyl)pteridine
To 2,4-diamino-6-hYdroxY-5-[4'-(3'-hydroxy-2',5'-dimethoxY-
2'-methYltetrahydrofuranYl)~pyrimidine (200mg, 0.664mmol) ob-
tained bY the Process described in Example 1, 6N hYdrochloric
acid (2.00ml) was added, and the mixture was stirred for 10
minutes at room temPerature. A Part of the reaction mixture was
taken, concentrated to drYness in vacuo and checked bY lH-NMR
sPectroscopy to confirm the formation of 2-amino-4-hYdroxY-6-
(I'-hydroxy-2'-oxopropyl)-5,6-dihydropteridine (see Example 5).
- 16 - 1 338 1 ~9
The remainin~ reaction solution was added to a susPension of
iodine (-6-OOms, 2.36mmol) in methanol (2.00ml) and methanol
(1.OOml) was further added thereto.
After the reaction had Proceeded for 10 minutes at room tem-
Perature, distilled water (5.OOml) was added. Then neutraliza-
tion was achieved with sodium carbonate (764m~, 7.21mmol), and
then distilled water (lOml) was adde~ to the reaction mixture to
obtain a dark brown solid that was collected by filtration. The
solids were washed with water followed by methanol and dried in
vacuo to afford 99.2mg (63.5X) of the titled comPound, as Pale
Yellow solids.
H-NMR sPectrum (DMSO-D6, ~ PPm) :
2.18 (3H, s, -OCH3)
5.24 (lH, d, J = 6Hz, Proton at 1'-Position)
6.33 (lH, d, J = 6Hz, hYdroxY Proton at 1'-Position)
6.96 (2H, br.s, amino Proton at 2-Position)
8.71 (~H, s, Proton at 7-position)
IR spectrum (K8r, cm-l) :
3248, 1652
mili-MS sPeCtrUm : CgHgNsO3,
235 (M+), 219, 192, 177, 163, 136, 122
Exa~Ple 10
L-BioPterin
To 2-amino-4-hYdroxY-6-(i'-hYdroxY-2~-oxopropyl)pteridine
(23.6ms, 100~ mol) obtained bY the Process described in ExamPle
9, in distilled water (lOOml), ~ -nicotinamide adenine di-
nucleotide Phosphate (reduced tYPe) (167m~) dissolved in 0.2M
PhosPhate buffer (PH 6.4, 100ml) was added. One unit of sepia-
Pterin reductase from rat erYthrocytes PartiallY purified bY the
method described bY Sueoka et al. [nBiochim. BiophYs. Acta" Vol.
717, Pa~e 265 (1982)] was added to the mixture. The resulting
solution was incubated for 5 hours at 30~ with shakins,
ultrafiltered, and freeze dried. The freeze dried material was
dissolved in a mixture of 0.1N acetic acid/MeOH (95/5, V/V) and
Purified by hish-performance liquid chromato~raPhy usin~ a
Develosil ODS column. The solvent was removed from the resultin~
- 17 - 1 33~ 1 49
L-bioPterin-containins fractions to obtain a residue which was
susPended in a small amount of distilled water and freeze-dried
to afford 10.5m~ (44X) of the desired comPound, as Pale Yellow
powder.
~ ]2~5 = -60 (c = 0.13, O.lN-HCI)
UV sPectrum (O.lN-HCI) A max nm
210, 247, 320
H-NMR sPectrum (DMSO-D6, ~ PPm):
1.06 (3H.. d, J = 6Hz, -CH3)
3.92 (lH, m, 2'-H)
4.44 (lH, dd, J = 5Hz, SHz, l'-H)
4.69 (lH, d, J = 5Hz, 2'-OH)
5.58 (lH, d, J = SHz, l'-OH)
6.86 (2H, s, 2-NH2)
8.71 (lH, s, 7-H)
11.42 (lH, br.s, 3-NH)
Said value of sPecific rotation is substantiallY the same as
that rePorted by B. Green et al. t"Chem. Ber." Vol. 99, pase
2162 (1966)3, and the values of UV and NMR sPectra coincide with
those of standard samPIes.
.. . . ... . .
