PTERIDIKECARBOXAMIDE DIURETICS.I
September 1966
17~-Methyl-3~-thiocyano-5~-androstan-l'7/3-01-2-one (X).--A solutioii of VIIId (1.0 g) in acet,oiie ( 3 5 ml) was treated dropwise with standard chromic acid solutio11.~5 The excebs reageiit wa': destroyed by a small amomit of isopropyl alcohol. The inorganic salts were removed by filtering through diatomaceous earth and the filtrate was concentrated in zlacuo, t,he residue was diluted with H20,and a crystalliiie product was collected. Recryatallization from methanol-I-120 afforded the an-thiocyanate X (0.7 g), mp 151-153°. An additional recrystallization from methanol gave an analytical sample: mp 15:3-154'; [ , ] ~ S D S13i.5'; x,,~ 292 mp ( e 60); rimr, 232.5-239.5 ( 3 p - R ) , 7 3 (C-17 methyl), 50.5 (C-18 methyl), 49 cps (C-19 methyl). 2~-Thiocyano-5~-androstane-38,17/3-diol (XI).-To a n ice-cold solution of Va (4.0 g) in T H F (100 ml) wvas added a cold solutioii
697
of lithium tri-~-butoxyalumiiiumhydride (20 g ) in T H F (100 ml). ~ for 1 hr a t about 5' and poured into an The reactioii w a stirred ice-cold 5 5 AcOII solution. The proditct was extracted with ether aiid the extracts were washed with HZO, 5% PiaHCOa, and finally H Y Oagain before drying over anhydrous Na2S04. Solvent removal in vacuo left a white solid which was recrystallized from acetone-hexane to give the diol X I (3.6 g), mp 202-205", [ C I ] ~ ~ D +23". A n d . Calcd for. CPOHSINO2S:C, 68.7%; H, 8.94. C, 68.57: I€, 9.23.
Found:
Acknowledgment.--We wish to thank Dr. F. B. Coltori for his interest and comments during the course of t,his work.
Pteridinecarboxamide Diuretics. I. Reaction of 4,6-Diamino-5-nitrosopyrimidineswith Substituted Malonamides' T. S. OSDESE,ARTHURA. SANTILLI,~ LEE E. NCCARDLE, ASD MARVIX E. ROYENTHALE Wytth Laboratories, Inc., Research and Development DivisLon, Raclnor, Pennsylvania
Receizled JIarth 12, 1966 The reaction of 4,6-diamino-S-riitrosop~rimidiiieswith a iiumber of ?i,S'-bis-substituted maloriamides in the presence of 1 equiv of sodium iii ethanol afforded mixtures of 4-amino-7-substituted amino-N-substituted 6pteridinecarboxamides and 4-amino-i-hydroxy-N-substituted 6-pteridinecarboxamides. The 7-substituted sminopteridinecarboxamides were found to be effective oral diuretics in rats, whereas the i-hydroxypteridinecarboxamides were inactive at comparable dose levels.
The base-catalyzed reaction of 4,6-dianiino-5-nitroso2-phenylpyrimidine (I) with cyanoacetaniide yields 4,7-diamino-2-phe1iyl-G-pteridinecarboxamide (la) (see Scheme I).3 Variation of substituents on the 2 position SCHENEI
I
NH?
I11
NH2
Ia XCCH C O N I XiOEt
I
", I
I1
of the pyrimidine, as well as substitution on the amide nit'rogen of the cyanoacetamide, permits the preparation of many biologically active substituted 6-pteridinecarboxaniides. These have been the subject of several recent patent'si I n each of these react'ions, ring closure to the pteridine results froiii the elimination of 1 equiv of water between the nitroso group of the pyrimidine and the active methylene group of the cyanoacetamide and the coricomit'ant addition of the amino group of the pyrimidine to the nitrile group of the cyanoacetamide. The reaction of I with diethyl malonate in the presence of 1 equiv of sodium in et'hanol affords ethyl 4aniino-7-hydroxy-2-phenyl-6-pteridinecarboxylate (11). In this reaction, ring closure occurs with elimination of mater and ethanol. Other exampIes of pteridine formation by reactions involving 4-amino-5-nitrosopyrirnidines have been described by Pachter and co-workers.j h recent review has appeared 011 biologically act,ive pteridines derived froni 4-aniiilo-~-riit,rosopyridine~.~ In view of what has been reported concerning these types of reactions an attempt was made to prepare 4-amino -7 - hydroxy-2 -phenyl-6-pteridinecarboxaniide (Ha) in a single step by the reaction of I with malonamide in the presence of an equivalent amount of sodium ethoxide. It was expected t,hat water and ammonia would be eliiiiinated in the reaction, thus (1) A preliminary account of this work u-as presented before the Division of Medicinal Chemistry, 150th National Meeting of t h e American Chemical Society, Atlantic City, N , Sept 1965, Abstracts, p 16. ( 2 ) T o whom inquiries regarding this article should be sent. (3) T. S. Osdene and E. C. Taylor, U. S. Patent 2,975,180 (IQ61). (4) J. Weinstock, U. S.Patent 2.963.478 (1960); T. Y. Osdene and A . .%. Santilli, U. S.Patents 3,138,595,3,138,591, 3,138,592, and 3,122,547 (1964). (5) I. J . Pacliter and P. E. Nemetli, J . Org. Chem., 28, 1187 (1963); I. J. Pacllter, P. E. Nemeth, and A . J. Villani, t h i d . , 28, l l Q i(1963). ( 6 ) T. S.Osdene in "Pteridine Chemistry." \V. Pfieiderer and E. C. Taylor, Ed., The Macmillan Co., S e a York, h-.I-., 1964, p 6 5 .
fi98
f g h I
i k 1 m n
SH. Is
699
PTEEIDINECAEBOXAIIIDE DIURETICS.I
September 1966
TABLE I1 7-HYDROXY-?uT-SLJBsTITUTED
6-PTERIDINEC.iRBOXAMIDES
I1
Mp, O C
Recrystn solventa
a
b
340" dec. 316" dec.
D-G E
18.1 27.1
C13H10N60~ Ci6H16N603
C
290
A
31.7
C I ~ H ~ ~ X L ; ~ 48.54 O ~ S 4 . 0 7 24.27 48.19 4.80 24.60
d e f
E A
12.9 4.0
g
279 253 248 310
A A
18.9
CiiHi4N603S Ci4H20N603 Cl&oN6Oa Ci7Hi~ClN603
42.57 52,49 58.69 52.51
h
292
A
18.5
CijHi6N603S
49.99 4.48 23.32 49.83 4 . 4 1 23.20
1
276
A
11.7
Ci7HisN602S
55.12 4.90 22.69 55.56 4.89 22.71
278-279
E
50.0
Ci~H21N703
57.71 5 , 3 5 24.80 57.52 5.02 24.37
320 >360 301 296 258-260
E E E E
16.8 12.0 15.4 33.3 13.6
C14Hi2~602 CijH14N6Oz Ci6Hi6N602 Ci7H18N602 CigH22N6O~
56.75 58.05 59.25 60.34 62.28
Compd
A
Yield,
70
Formula
7---
C
------'A
% calcd--
C
"
found---H Pi
55.31 3.57 29.78 55.54 3 . 6 6 29.34 56.46 4.74 24.70 56.26 4.52 24.34
4.55 6.29 5.47 4.41
4.08 4.55 4.97 5.36 6.05
27.08 26.24 22.81 21.62
28.37 27.08 25.91 24.84 22.94
42.70 52.42 58.87 52.21
56.51 57.95 59.11 60.09 62.35
4.69 6.29 5.63 4.52
4.20 4.55 5.00 5.14 5.97
26.76 26.13 22.49 21.53
28.18 27.30 25.76 24.76 22.63
TABLEI11 ?;.~'-BIssUBSrrITUmDl I d L O N S M I D E S , CH*(CONHR)z Compd
I11
RIp,
O C
Recrystn solventa
N
38.0
49.53 8 . 3 1 12.84
49.71 8.31 12.61
79.1 22.4 51.0 31.0 50.0
CiiHzeN204 C13H26N204 CllHZ2N2O1S2
5X64 9,OO 11.37 56.91 9.55 10.21 47.45 7.97 10.06
53.37 8.95 11.24 56.93 9.36 10.20 47.83 7.93 9.90
CijHzN204
61.20 7.53
61.09 7.52
117 80 120
B
B F
h
123-125
B
15.5
i
100
B
36.6
C
N
found-? H
C~HiaNz04
d e f g
H
C
Formula
F A
C
.-----%
70
91-92 128-130
a b
___-% calcd--7
Yield,
9.32
9.59
40.0 129-130 B 23.0 148 F 37.6 139 F 1 71.9 131 F m ClSH30x202 66.62 11.18 10.36 66.46 11.16 10.23 81.4 129 B n AtSee footnote a of Table I. IT. H. Rauscher arid W.H. Clark, J . Am. Chem. SOC., 70, 438 (1948), reported mp 127-127.5'. Burroughs Wellcome and Co. (U. S. A , ) Inc., British Patent 701,209 (1953), tempts to recrystallize this material were unsuccessful. 73, 1967 (1951), reported mp 120.5'. f RI. Freund, reported bp 160" ( 2 mm). e R. W. Bost and L. 5'.Nullen, Jr., J . Am. Chem. SOC., 359 (1921), reported mp Ber., 17, 133 (1884), reported mp 128'. 0 J. \*. Backes, R. W. Weat, arid ?If. A. Whiteley, J. Chem. SOC., 149". * Lit.B mp 139'. * Lit.g mp 132.5'. j
k
the compound being tested. This dose of test compound was given to 8 rats (T). An osmotic diuretic urea solution was given concurrently to a separate group of 8 rats (U) a t a dose of 960 mg/kg. The animals were placed in metabolism cages, 2 rats/cage, and urine was collected for 5 hr. Volume was measured and sodium and potassium were determined.' The results in Tables IV-VI1 are expressed as the average
ratios of urine volume and sodium levels in test animals (T) to those in urea-dosed animals (U).8 All ratios greater than unity for volume and sodium output represent statistically significant diuretic responses. Structure-Activity Relationships.-Table IV gives the diuretic activity of a number of 4,7-diamino-Bpteridinecarboxamides having various substituents on the N atoms of the 6-carbamoyl and the 7-amino
(7) Uaird .\tornic Model KY-1 tlame photometer adapted for automatic [ion-tylle analysis.
(8) \V. L. Liiiscliitz, Y'herup.. 79, '37 (1943).
Z. Hadidian, and .\. lierucsar, J . I'hurmucol. E ' r p t f .
TOO
RHNOC
(IO) Aleitinn poititS x e r e riererniinwl uti a Tliomas-Hoo\ e r capillary melting point apparatus and arr i,orrrrted. I-ields reported in Tables 1-111 are the results of single experiments a n d should not be constrned as rnaxinial. Im. Ciirm. Soc.. 81, 2-112 : 11) t;. C. Taylor. 0. Voel, nnd C'. I-. Clicng,
,
1'JSDI. (12) i-iil,iiliad I,) . \ r a ~ ~ h u('iieniicuia. e In360". This material was set aside for separate treatment as described below for the preparation of IIk. The mother liquor was concentrated zn uacuo. Ethanol (5 ml) was added to the residue, giving Is as a yellow crystalline product (1.0 g), mp 253'. The analytical sample (mp 258') was obtained by recrystallization from ethanol. 4-Amino-7-hydroxy-N-methyl-2-phenyl-6-pteridinecarboxamide (IIk).-The 3.3 g of material, mp >360", obtained as described above was dissolved in 400 nil of water. The water eolution was treated with charcoal and filtered, and the filtrate A R S acidified with 10 ml of glacial acetic acid. A yellow precipitate (mp 315') was deposited and recrystallized from 2-ethoxyethanol, affording IIk (1.5 g), mp 320". 4,7-Diamino-2-phenyl-6-pteridinecarboxamide (Ia).-To a solution of 0.2 g of sodium in 250 ml of absolute ethanol was added 4.3 g of I followed by 2.0 g of malonamide. The reaction mixture was stirred and heated under reflux for 0.5 hr, yielding Recrystallization from I a as a precipitate (3.0 g), mp >360'. aqueous dimethylformamide afforded a product with an infrared spectrum identical with I a prepared from I and cyanoacetamide as previously described.3 4-Amino-7-hydroxy-2-phenyl-6-pteridinecarboxylic Acid (III).-To a solution of 0.8 g of sodium in 500 ml of absolute ethanol was added 6.45 g of I followed by 5.6 g of diethyl malonate. The mixture was stirred and heated under reflux for 20 min, during which time a precipitate was deposited. The reaction mixture was cooled in ice and the product wa> removed by filtration, dissolved in boiling water, and acidified with acetic acid. The resulting precipitate was recrystallized from aqueour dimethylformamide, affording I11 as a monohydrate, mp 276' dec. Anal. Calcd for Cl3H9NjO3.H20: C, 54.73; H, 3.89; N, 24.55. Found: C, 54.37; H, 4.03; N, 23.94. Ethyl 4-amino-7-hydroxy-2-phenyl-6-pteridinecarboxylate (11) was prepared in identical fashion but 1 equiv of sodium was used. From 4.3 g of I and 3.5 g of diethyl malonate in 100 ml of absolute ethanol containing 0.5 g of sodium was obtained 5 g of 11. The analytical sample (mp 255') was obtained by recrystallization from aqueous dimethylformamide. Anal. Calcd for Cl6Hl3N6O3:C, 57.87; H, 4.21; N, 22.50. Found: C, 58.02; H,3.85; N,22.76. 4-Amino-7-hydroxy-2-phenyl-6-pteridinecarboxamide (IIa).A mixture of 1 g of dried I11 and 35 ml of SOC1, was boiled under reflux for 2 hr. The SOCl, was removed zn uucuo on a rotary evaporator and the residue was treated with 20 ml of 3 N NHIOH. The mixture was heated on a steam bath and filtered, and the filtrate was acidified with 3 3' acetic acid. The precipitate thus obtained was recrystallized from aqueous dimethylformamide, affording I I a (0.18 g), mp 340" dec. Anal. Calcd for C13H10N602: C, 55.31; H, 3.57; N, 29.78. Found: C, 55.54; H,3.66; N,29.34.
Acknowledgment.-The authors wish to thank Dr. Gordon Ellis and staff for the microanalytical determinati ons.
