,July 19GG
62 1
SOTES
TABLE I ARYLAZOPYRIDOXINE HYDROCHLORIDES ---Calcd,
%-
6 sitbstitiient
C
n
Phenylazu p-Chlorop henylaxo p-Tolylazo p-Xi trop heiiylazo
61.5 48.9 55.6 47.3 47.6
5.8 4.7 5.6 4.2 4,3
~
'l
~
l
l
~
f
~
J
l
~
~
~
~
~
~
a
s 15.4 12.2 13.0 15.8 11.9
----Found, C
61.7 49.0 56.0 47.6 47.3
n
yo--N
5.5 .5 . 1 6.4 4.4 4.7
15.3 11.9 12.7 15.4 11.6
Xot hydrochloride. 4.45
CH,OH
j.82
4.70
CH,OCOMe 7.51
carried out, but disclosed.
110
significant biological activity was
G e C 0 0 C H 2 , f f C O M e 7,98 H
0.5
I
Experimental Section
G/ I
Me
Me
I-
7.20
5.29
L\7
I1
test,'!* but crystalline derivatives have not previously been reported. Hydrogenation or dithionite reduction of the phenylazo conipounds gave 6-aminopyridoxine which was characterized as the tetrabenzoyl derivative. The structure of these compounds was confirmed by the nmr spectra in trifluoroacetic acid which showed no peak due to a ring proton in the 6 position. Triacetoxypyridoxine gave the methiodide (I1; nnir spectrum on 'T scale) ; attempts to condense this with aromatic aldehydes were unsuccessful. Pyridoxine mas condensed with ethyl bronioacetate, arid also the Soxide of the tribenzoyl derivative of pyridoxine was prepared. These and some further compounds3 were tested for in viilo inhibition of Sarcoma 180 tumors in mice (see Table 11). Antimicrobial tests were also TABLE I1 CORIPOUXDS TESTED FOR INHIBITION OF Q.\RcoM.\ Compd
6-Amino-4,5-bishydroxymethyl-3-hydroxy-2-methylpyridine.
A.-6-Phenylazopyridoxine (1 g ) in ethanol (150 ml) was shaken Pd-C (0.2 g ) a t 20". Hyunder 3 atm of hydrogen over drogenation was complete in 3 hr; steam distillation followed by acidification v i t h 12 -I' HC1 gave the amino derivative (0.2 g, 32%) which crystallized from ethanol as microcrystals, nip 180-186' dec. Anal. Calcd for C8H,sC1S203:C, 43.4; H, 5.9; S, 12.7. Found: C, 43.6: H, 5.9; X, 12.7. B.-Sodium dithionite (20 g ) was added to the alkaline azo solution (prepared from 10.3 g of pyridoxiiie hydrochloride) a t 75' with st,irring. Aft,er cooling, anilitre was extracted with ether. The aqueous residue was acidified with 12 S HC1, kept 2 hr at 50", filtered, and evaporated to dryness to give 6aminopyridoxine hydrochloride (7.3 g, 66Yc). A n d . Calcd for CsH&lS,Oa: C, 43.4; H, 5.9; N, 12.7. Foimd: C, 43.5; H, 6.0; K,12.i. Benzoyl chloride (3.5 g) and 2.5 S NaOH (30 ml) was treated with 6-aminopyridoxine hydrochloride (1 g) t o give the tetra180 TUMORS benzoyl derivative which separated from methanol as pale yellow crystals, mp 86-88'. Activity" Anal. Calcd for C36H28N207: C, 72.0; H, 4.7; K,4.7. Found: C,71.6; H,4.4; N, 4.5.
2,4,5-Trihydroxymethy1-3-hydroxypyridine 3-Hydroxy-4,5-bis( hydroxymethyl)pyridirie-2aldoxime hydrochloride N-Hydroxypyridoxiiiium chloride j-4cetosy-3,4-dihydrox~methylpyridine-2-aldehyde S-oxide p-dimethylaminoaiiil 3-E thoxymethyl-4-acet oxymethyl-5-hydroxy-6methylpyridine K-oxide 3,4-Dihydroxymethyl-5-hydroxy-6-aminumet hglpyridine dihydrochloride Pyridoxinium-1-acetic acid 2-( N-Pyridoxiriiiim )acetophellolie 6-Pheiiyla~opyridoxiiiehydrochloride 6-( p-Chlorophetiylazo)pyridosiiie hydrochloride 6-Tolylaxopyridoxine hydrochloride ib p - [2-(3,4-l)ihydroxymethyl-5-hydroxy-6-methyl)pyridylazo] pheiiyldfoiiic acid hydrochloride 2-Chlor0-3,4,8-triacetylpyridoxine -C 2-Amiriopyridoxine hydrochloride P)-ridoxine K-oxide hydrate Compoiinds were tested a t 500 mg/kg. * O n e test only. Segative at 2Fj0 mg/kg (insiifficieiit) material for test at higher dosage).
(1) .IItiba . and Ii. Miti, Sci. Papers Inst. P h y s . Chem. Res. (Tokyo), 34, 1014 (1988); Cliem. Abstr., 33, 205 (1939). (2) Z. Deyl and J . Rosmus, J . Clmmatog., 8 , 537 (1962). ( 3 ) G. R . 13edfiiri1, .\. R . Iiatritzky, and H. M. \Vuest, .I. Cliem. S o c . , 4600
(1963).
6-Phenylazopyridoxines.-The diazonium solution (from 9.3 g of aniline and 100 ml of 6 S HC1) was added a t 5-10" with stirring to pyridoxine hydrochloride (21.2 g ) in u-ater (500 nil). Sodium hydroxide (2.5 S )was added simultaneously to keep pH 8. After stirring for a further 1 hr a t 20", 12 HC1 was added to pH 4, and the precipitate --as filtered o f f and dried i n a vacuum desiccator. Substituted compounds were prepared analogously. The products (Table I ) crystallized from dilute HCl.
3-Acetoxy-4,5-bisacetoxymethyl-l,2-dimethylpyridinium Iodide.-Pyridoxine triacetate ( 5 g ) and methyl iodide (15 ml) were refluxed for 12 hr in methyl cyanide (100 ml). Evaporation of the solvent and recrystallization of the residue from ethyl acetate gave t,he methiodide (7.4 g ) as yellow plates, mp 114114.5". Anal. Calcd for CljH?,INO6: C, 41.2; H, 4.6; S, 3.0. Found: C,40.Y; H, 4.9; N,2.9. 1-( Ethoxycarbonylmethyl)-4,5-bishydroxymethyl-3-hydroxy2-methylpyridinium Bromide.-Pyridoxine ( 5 g ) and ethyl bromoacetate (6.0 g ) were refluxed 2 hr in ethanol (20 nil). Cooling gave the bromide (6.2 g, 62%) which separated from ethanol in needles, mp 128.5-129'. .Anal. Calcd for C1ZF118BrSOj:C, 42.9; H, 5.4; N, 4.2. Foinid: C, 43.0; H, 5.6; N , 4.1. S m r spectriim (in I),0 with (MeaK++)rS042-as internal qtandard a t 6.81): 1.71 (6-H), 4.30 (4-CH4), 4.89 (S-CH,), 5.15 (N-CH,), 5 . i 5 (quartet)(CHz of Et), 7.35 (2-CH3), 8.70 (triplet) (CH, of Et). 3-Benzoxy-4,5-bisbenzoxymethyl-2-methylpyridine 1-Oxide.Pyridoxine tribenzoate4 (1 g ) was treated with monoperphthalic acid ( 2 g ) in chloroform (10 ml) and ethanol (10 nil) for 4 days a t 20". The mixture was shaken with saturated aqueous SaHC03, dried (MgSO,), and evaporated to give the oxide (0.8 g, 76Yc), which crystallized from benzene as needles, nip 133-134". (4) A . Itiba and K. hliti, Sc7. Papers I n s f . Phip. Chem. Rrs. (Tokyo), 35, 5 3 (1038), C h r m . .Ibstr., 33, 3430 (1939).
Studies on Latent Derivatives of Aniinoethanethiols as Potentially Selective Cytoprotectants. V. Syntheses of S-(2-Aniinoethyl) Ifethyl Hydrogen Phosphorothioate'
I, 1; 11, 1: 111, 1:
= = =
11
CIl, C(. therapeut i r agents with corisequent incw:ise i l l t1icr:ipcwtic. effect in rmcer. T i i t , sodiuni s:ilt of the parent compound cysteapiiic 8-phosp1i:ttc (I) ivas prepared by the iiiethod of ,lker-
fcltlt, involving the rewtion of 2-bronioethyl:tmiii(~ hytlrobroriiidc with trisodiurii phosphorothioate, the 1:ittc.r tieing obtained by the rcnc%iori of sulfur wit 11 1Y"l followrtl hy treatincrit of tlic proiluct with sodimii Iiyilrouid~. h
I Y Y 1--+ ~ l'SC1, \ICli
XaOlf
-+PSiOS:r)1
I3rCilL!II+Ilf~~I W r
+I
\\-(a origiii:illy tried t o syntlicizc 111by an :malogou> route. Thiophosphoryl chloride c t h:u io 1 t o y i cld 0 - e t hy 1 p l i o q h ) Hydrolysi5 o f the latter with rotliurii hydroxide, lion-ever, tailed to yield the desired disodiurii salt (IVaj. T h c 0-methyl :tiinlog (11%)had been prepared? in 1911
PSCl,
(RO)PSCl, +-
(RO)PS(ONa),
IVa, R=C2Hj 11, R=CH11) Slipuorted by Research contract PII43-62-170, Cancer Chemotllerapy Kational Service C e n t e r , Xational Caiicer Institute, Kational Institute- o f Healtli. 13etliesda, lid. ( 2 ) 3. .\kerfcldt. A d a C h u m . Scand., 13, 1479 (1969). [:$I H. K. Yauiida and J . I,. Lambert, .I. .Am. C h e m . Soc.. 76, 5326 (1515.%,. i 1) \ \ . C . l.:mmrtt a n d 11. 0. Jones, .I. Chpnz. Soc., 99, 71.5 11911).
