Xovember 1969 HYPOCHOLESTEROLENIC TETRAZOLEH

&SUBSTITUTED 1001

The residual solid, mp 110-120", [ a l z 1 D +10.7" (c 1.0, H*O),
was crystallized from MeOH uiit'il the rotation became constant.
Thus 29 (6.6 g, 45%) was obtained. The free base (+)-1 (31))
(+)-1 .HC1 (33), arid (+)-1 hydrogen ( - )-t,artrat.e (35) were
obtained by conventional methods.
( - )-1 Hydrogen ( - )-Tartrate (30).-The retaiiied filtrate arid
the crystallizatioii mother liquors from the preparatioii of 29 were
combined and evaporated. S a O H ( 2 S)aiid saturated aqueous
KaC1 (100 ml) were added and the mixture \vas extracted with
Et,O (130 ml, three times). The extract yielded optically impure
(-)-1 base, [ a ] * ' D -30" (c 2.2, EtOH). This (-)-1 base (13.1
g) in NeOH (60 ml) was added to a solution of ( - )-tartaric acid
(10.8 g) in hIeOH (60 ml) a t room temperature. The solid
which separated, mp 131-154', [ ' Y ] * ~ D-36.5' (c 1.06, HzO), was
crystallized from AIeOH until the rotation became constant.
Thus 30 (6.6 g, G(,';) was obtained.

Hypocholesterolemic 5- Substituted Tetrazolesl

1tOl;ALD L.BT;CH,kN.k??, YILJIARS SPRASCIIASIS, A N D RICHARD h.1 ' A R T l x 4
Ifcseatcli, Division, Bristol Laboratories, Divzsion of B r i d o l - M y f r s Company, Syracuse, Al-euqYolk 13201

Received March 17, 1969

h iirimber of 5-aryloxyalkyl-, 5-arylthioalkyl-, and 5-anilinoalkyltetrazoles. along with a few other related
5-wbstituted tetrazoles, were synthesized by standard methods. A novel tetrazolylethylatioii reactioii was used
to byntheaize 5-[~-(3-chlorophenoxy)ethyl] tetrazole (68). In
tetrazole (53) aiid 5-[~-(3-~hloropheiiylthio)ethyl]
general, the 5-arylthioalkyltetrazoles provided the be3t combination of high hypocholebterolemic activity and
lnw toxicity.

It is well known that 5-substituted tetrazoles and The bicyclic dihydrobenzofuran and benzothiopherie
their carboxylic acid analogs have comparable dissocia- derivatives 111 arid IV (compounds 86 arid 87 in Table
tion constants.2z3 I n some cases this physiochemical 111) were also prepared.
analogy has been reflected in similar biological ac-
t ivit ies. -$
I n connection with other studies, it was discovered
that 3-aryloxymethyltetrazoles (I) were inhibitors of
cholesterol biosynthesis from acetate-l-'*C z" vitro. lo
l~ollow-upi~ vivo studies revealed that these compounds, I11 Iv
although somewhat toxic, lowered normal serum choles-
The object of these syrit'hetic modificatioiis was to
H obtain compounds which combined potent hypocholes-
A~-O-CH~~N,N I terolemic activity with low toxicity. This paper
1
I describes the preparation and some physical properties
E-N of t'hese 5-substituted tetrazoles along with the pre-
I liminary hypocholesterolemic screening data.
terol levels in rats. The plant growth hormone activity Chemistry.-In most cases the syntheses irivolved
of some compounds of type I has been reported by the preparation of the requisite nitriles (Table 11),
1Ic1lanus and Herbst.: I n view of the similarity of which were then converted t o the desired tetrazoles
these compounds to the known serum lipid lowering (Table 111) by standard methods.
agent ethyl a-(4chIorophenoxy)isobutyrate (clofi- Where carboxylic acids \yere used as starting ma-
brate) , I 1 whose active principle is the corresponding terials, conversions to the corresponding amides (Table
carboxylic acid, a number of compounds of general I ) were effected by ;iccepted methods. These amides
structure 11 were synthesized. were theri converted to nitriles (Table 11) either by
vacuum distillation from l'?Ojl? or b y reaction with
PoC13 arid
The aryloxy- arid arylthioacetonitriles were prepared
by refluxing the appropriate phenols or thiophenols with
chloroacet,otiitrilein w slurry of I<?CO3in acetone5 or in
branched chain). S H SaO;\le-11eOH.13 Homologs (straight or branched
1: = H , CH3 chain) were prepared by base-induced coridensa t'1011s'
(1) Some of these compounds have been described by R. L. Buchanan and
of the phenols or thiophenols with bromoalkylnitriles
8. .IPartyka,
. U. S.Patent 3,337,676 (1967). or acrylo~iit~rile'~ (Scheme I ) .
(2) F. R. Benson, Chem. Reo:..41. 1 (1947).
( 3 ) R. AI. Herbst, "Essays in Biochemistry," John IT-iley and Sons, I
SCHEME
Inc., New Tork. X. T..1966.. .D 141.
li. 11
(4) C. r a n d e Westeringh and H. Veldstra. Ree. Trav. Chim.,77, l l O i
KaH
(1958).
( 5 ) .J. AI. McManux aml 11. A I . IrerIJsl, J . O r g . Chvm.. 24, 1464, (1959).
ArXH + Br&"C€12),CS & ArXAH(CH2)nCX
DAIF
(t;) J. I < . l-2lwuod. K . M. l i e h a t . nnd C.i. L. liilyour. ./. B i d . Chew., ado,
W 7 3 (19651. ti li, CHI. CsHj; S 0, S: II = 0, 2
( 7 ) €3. Bruuwer-vau Stranleu, U. Solinger, c'. vnu de \\ edtrriugli, axid t j ,
V&lstra. K e f . Trau. Chim.. 77, 1 1 2 ~I 1958).
(X) ( > .F. Holland and J. N. Prrrirn, J . .\lei/. C ' / Z ~ I I L10,
, , 149 (1967).
( U I P. F. Juby, T. IV. IIudyma, a n d M. Broivn, ibid., 11, 111 (1968). (12) 11. IC. Kent and 8. 31. AIcl~;lvain,"Ornanic S>ntllrses," Cull. V d .
(10) Cnpublished results. 111, John \Tiley and Sons, Inc., New York, N. Y.,1Y55, p 493.
(11) .\tromid-S'8'. CPIB; .J. hl. T h o r p and TI-. S. Waring. S u t u r e , 194, (13) R. Dijkstra and H. J. Backer. Rer. T r m . Chim.,73, ,569 (lY54).
11-18 (1Y62), a n d many subsequent gapers. (14) S . .I.Heininger (to hfunsanto). U. Y. Patent 2,81Y,2Yl (1958).
I002

?'tie aiiiliiioacetoiiitriles were prepared by treating

the appropriate anilines jvith formaldehyde bisulfite
followed by KCS.'" 4-Chlorophenyl thiocyanate (21)
atid ,8-(2-chlorophenyl)propioriitrilr (43) were preparecl
:LY previously described.16si7
Thv nitrile. ~ w r rcoiiverted t o t etrazoleh by treat-
ment with S H 4 S 3iii DIIl;l* or with -11(S3)J in THIJ.t'i
The latter method proved to be the o ~ i eof choice iii the.
l)rrparntioii of I' (58, Table 111). Whcit SH4S3-DA\Il'
\vas used, 1-w i s susceptible to cleavag~by azide, givilig
tlic azidotetr:tzolc VI its the major isolatcd product.
Sovernber 1969 1003

TABLEI1
KITRILES

E-ield, Recrystn 1111 01 ilp

It, Hi It, x Y 7bletliud yo solventa (mm) O CI Formula Analyses
11 II 11 0 CII, ,1 74.3 72-78 ( l ) t CrH;NO
II II C1 0 CHI A 90.6 A-B 4-47' C,H&I?U'O
H CI 11 0 CII, A 68.2 89-93 C,H,CI?;O
(0 2-0 3)'
12 c1 €I I€ 0 A 31.3 102-106 (0 3 ) L CSHGClNO
1:3 C1 C1 H 0 A 99 B 8!5-881 CsH,C12NO
14 II CF, IT 0 A 84.8 81-8.5 (0 1 ) C9HfiFaN0 C, €I, N
13 H C1 II F 33 6 lW-108k Cy HaCINO
(0 1 )
16 II e1 €I 0 E 62 2 113-120 CioHioCINO

17 I1 1I c1 0 C 1.1(CH, ) B 91.1 -- (0 07)

I .IF80 CyHsCISO e, H,c1, s
(0 li-0 17)
I8 I1 C1 I€ 0 E 67.1 83-88 (0 1) c,II,ClsO c, I{, N
19 II C1 I1 0 E: 26.5 76-82 CioHioClX0 H , C1, S ; Cc
(0 1.5-0 2 )
"0 Ir 11 ci 0 B 81 . ( J 78-80 10 4 ) C I ~ H I ~ C I S O C, H, C1, N
21 1% II c1 S H 64.9 1 CiH4ClNS
22 II II €I S CHy 1) 83.9 118-123 rn CsHiKS
( 0 4)
23 II 1-1 CI S c H, -4 82 3 c: 84 .i-87" CsH,CINS
24 I1 CI II S crI, A T O .3 1)-A .50-32 CsH,CINS e, 11, c1, N, s
2.i C1 II II 8 CH, A 79 8 116-12.5" C,H&IXS
(0 2.5-0 3 3 )
"6 S C€I? 84-92 (0 1 ) C,TI,FNS c, 11; Nd
27 S CII, 83 .5-8.i .5, 1.58- CaH8N2028 c, H, N , S
170 (0 1-0 2)
28 H 11 CH,0 8 A 97. 4 1,n CqHSKOS
29 H CF, H 8 A1 73.3 83-94 CyHfiFJYS C, H, N
10 03-0 3 )
30 H C1 I1 8 E 75.7 129- 132 CioHioClNS C, H; Ye
(0 07)
31 If li C1 8 B 87 A 46-47, CgHdCINS e, €I, s
9.5-97 (0 3 ; )
:3 2 E 62.4 80-8.5 ( 0 03) C,H,CINS c, I I , N , s
3:j E 65.3 82 .i-86 ( 0 1) CyHEClN8 C, H j C1, s,8
:34 E 77 -5 !)8-10:3 ( 0 1.5) CIoH,,NOS c, II, x; S'
33 B 81 4 -4 51 .5-33 .i, CioHioCINS 11, c1; C,u N"
87-92 (0 13)
36 H II Br S B 83 A ,50-,52 CI,HIoBrSS C, H,?;
37 H H H N II G 64.7 130-1 400 CZHXSZ
(0 33-0 3 )
38 H H CI NH CHI G 62.9 E-A 67-68 .i c81r,cis2 C, H, C1, S
39 H C1 1-1 KH CH, C: 31.5 148-1.52 CrH;CIS> C, H, C1, S
(0 25-0 i 3 )
'40 s11 CII, 97-100 Cs€I;NaOl c, H,s
41 c, 11, N
42 c, 1-1, N
43

44

43 C 73.9 86-872 ( 0 . 2 ) CgH;KO

C r 1 C,H,CISS

A, Skellysolve B (bp 60-80'); B, 100% EtOH; C, CPrOH; D, CHCI,; E, EtOAc. S : calcd, 7.16; foimd, 6.54. C : calcd,
61.39; found, 60.87. N : calcd, 8.38; found, 7.93. e N : calcd, 6.62; found, 6.06. f S: calcd, 16.59; found, 13.79. 0 C : calcd,
56.73; found, 56.28. * S : calcd, 6.62; found, 6.17. i Pee ref 5 . i A. Campbell (to Parke, Davis & Co.), V. 8. Patent 3,139,435
(1964). k H. Ufer (to I. ( 2 . Farben. A.-G.), German Patent 670,357 (1939). 1 Used in the critde state. rn See ref 13. G. H. Hitchingi,
b:. A. Falco, and €3. Roth (to Burroii~hP-~ellcoIne arid Co., U.S.A.), U. S. Patent 2,933,367 (1960). 0 See ref 15. 11 Pee ref 17. q A I .
Julia, Bull. SOC.Chim. Fr., 1363 (1956). Kot determined.
100-I

1
I

E' i.; 110: I 1

1.'
I
I) K
I
\
\
I
.\ 111 I
.\ I
(*
 HYPOCHOLESTEHOLEMIC TET~~AZOLES
&SUBSTITUTED 1005

TABLE 11-
HTPOCHOLESTEROLEMIC
ACTIVITY
OF ~-S:UBBTITVTTED
TETRAZOLES
Dose, Serum cholesterol, Dose, Serum cholesterol
N0 mg/k % change No mg/ kx Yc change
47 400 10
(Toxic)<&
+ 6b 400
400
- 44
- 25
4X 400 69
100 - 14 i0 400 - 32
49 400 (Toxic) 71 400 -31
100 - 26
.i0 400 (Toxic) "2 400 - 20
100 -6 "'I
13 400 -6
31 400 (Toxic) 74 400 --h
m-
100 -21 i.I 400 0
52 400 - 38 76 400 - 11
. 3i 400 - .;2 77 400 - 24
100 -7
34 400 - 36 73 400 - 19
_-
.).) 400 - :<3 70 400 - 20
56 400 - 13 so 400 - 12
-m
.)I 400 - 26 $1 400 f 2
;. h 400 - 13 52 400 - 12
39 400 -8 '13 400 - 17
60 400 +3 $4 400 - 3x
61 400 - 40 -h.i :300 +3
62 400 - 36 46 400 0
200 - 24 '17 400 - 20
63 400 - 22 b8 400 +x
64 400 - 17 89 400 + 13
63 400 - 1s CPIB 400 - 25
66 400 - 24 200 -l h
67 400 - 31
0 Ilrug-ielated death of at least one of the test animals.

analyzer method (S24AP) and are recorded in Table Experimental Section

IV. I n general, compounds that lonered serum choles-
terol by a t lenst 2OyO\\-ere considered act'ive
In general, the most active compounds are the 5-
aryloxymet hyl- aiid bnrylt hiomet hyltetrazoles, the
latter being somewhat less toxic. An electronegative
substituent on the berizerie ring is necessary for optimal
activity (compare 61 with 60 and 66), but no direct
relationship between the degree of electronegativity of
the substituent' and hypocholesterolemic activity was
observed. Lengthening of the carbon chain to Ca
between the hetero atom and t,he tetrazole ring results
in 110 loss of activity. I n the oxygen series, chain ex-
tension also loners toxicity (compare 49 with 53 and
54). >lost of the compounds which incorporate a
single met'hyl or ethyl group on the carbon adjacent to
the tetrazole ringretain activity ( k .55,
, 57, 70,71, aiid
72), whereas dimethyl analog* 58, 74, arid 75 are nearly
inactive. Alkyhtioii of the tetrazole ring as in 88 and
89 destroys activity, indicating that an anionic center
is required. Oxidation of one of the thio analogs (61)
to the corresponding sulfone (76) resulted in substan-
tially reduced activity.
5-(3-Chlorophenylthiomethyl)tetrazole (62) gave con-
sistent results in dose-response studies and was selected
for further evaluation.*2 I t had a minimal effective
dose (20% lowering of serum cholesterol) of 200 mg/kg,
with a n oral LDjo of 2200 mg/kg in rats. From Table
IV it' can be seen t'hat compound 62 had activit,y com-
parable to the standard drug clofibrat'e (CPIB) in this
assay.
( 2 2 ) (a) J l u r e <letaile,l account.s of tlie pllarmacology and Iiiocliemirtry
of 5-(3-c1ilorophenyltIiiometh~1)tetrazole\rill he reported else\\-liere. ( h )
Initial clinical findings have been reported hy D. T. Xash, L. Gross, IT-.
Hair, and li. .\are, J . Clin. Pharmucol., 8, 377 (1968).
The melting poiiit,s were obtained in capillary tiibeli with
a Thomas-Hoover Ui~i-Melt apparatus and are uncorrected.
1Vhei.e aiialysea are indicated only by symbols of the elements or
fiinctioiis (Tables I-111), analytical reeiilts obtained for t,hose
elements or fiiiirtioiis were within 1 0 . 4 5 of the theoretical
valiier. Xmi. spectra (CDC13) were obtained iisiiig a Variaii
Associates 31odel A4-60spectrometer. Chemical shifts ( 6 ) were
measured downfield from TlIS.
Amides (Table I). Method A.-Compounds 1-5 aiid 8 were
prepared by procedures similar to the one described by Kent aiid
McElvaiii for the preparation of isobiityi,amide from isobutyric.
acid.":' >-Chlorobenzo[b]thiopheiie-2-cart~ox~-licarid was pre-
pared by the method of Campaigiie aiid Clitie for beiizc)[h]thio-
pheiie-2-carboxylic acid. 2 4
Method B.-Compoiiiid 7 was prepared as desci,ibed by
Stoermer aiid KBiiig.*Z
Method C.-Knoeveiiagel's method fur the preparation of
pheiiylamiiioacetoiiitrile,'~ wheii used i i i the s y i i hehis
~ o f thc
:<-riit,roanalog, resulted in t.he amide 6 ( l i < ' L )aloiig with I hr
desired iiitrile (40) (lO.Syo).
Nitriles (Table 11). Method A.-Compounds 9-14, 23-29, 42,
aiid 44 were prepared by procedures similar to the oiie described
by AIcAIaniis aiid Herbst for the preparation of pheiioxyaceto-
iiitrile (9).5
Method B.-Compoiiids 17, 20, 35, aiid 36 were prepared by
the method of Teagiie aiid Short for the preparatioii of iiirotiuu-
nitrile. 26
Method C.-Compoiuids 40, 45, and 46 were prepared as
described by Jiiby, et al., for the prepai,atioir of 2-(2,6-dichlo1o-;2-
met hylaiiiliiio)be~i~,oiiit~ile.~
Method D.--<:~~ii~p~iiiitl 22 was prepared :tcw)rdiiig t o the
procedure of Uijkstra and Backer.13
Method E.-Compounds 16, 18, 19, 30, and 32-34 were
prepared by the method described by Genzer, et al., for the
( 2 3 ) R. E. Kent and S. hI. hIcElrain, "Organic Syntheses," Coil. Vol.
111. .lniin JVilry and Sori.*. l n r . . Xmv York. N . Y,, 145.5, p 490.
( 2 4 ) 1,:. ('ani[,aigne and 11. IC. ('line, .l, ( J r y . Chf'm,,21, 92. :i$l (1!15ti),
( 2 5 ) K.Stoermer and IV. lionig, Ber.. 39, 492 (1906).
(26) P. C . Teague and JV. A. Short. "Organic Syntheses." Cull. Vol. I \ - ,
John \Vile?. and Sons. Inc., S e w l o r k . S . I-.. 1963. p 7 0 6 .