arid 25 g of phenol was heated and stirred on a steam bath for 4 hr. After cooling, the mixture was treated with 100 ml of 31e2C0 cont,aining 10 ml of concentrated I-ICI, and the resulting precipitate was collected by filtration. Two recrystallizations from iPrOII gave 2.3 g (24YG) of 6-chloro-2-met,hoxy-9-(piperidinriamino)acridirie monohydrochloride (JTIIf) as i i bright yellow powder, mp 204-206" dec. 4,4'-( 1,4-Piperazinediyldiimino)bis(7-chloroquinoline) (VI1j. 4 mixture of 19.8 g (0.10 mole) of 4,7-dichloroquinolirie and 5.8 g (0.050 mole) of 1,4-diaminopiperaziiieg was heated in 30 g ( i f phenol for 4 hr oii a steam bath. Excess roncent,rated llCl Tvas added, and the mixtiire was poiired itito 3 I. of 11erCO. Tritmation with fresh MerCO, theii m-ith EtZO, gave 9.2 g uf yell^^^ d i d , mp >300". This was dissolved i t i I-{&, and the sollition was made alkaline with SaOH. The resulting precipitate \\-as collected by filtration, washed well with FI&, :md dried a t 63" in vacuo to give 2.5 g (115%) of a pale ye110~-solid. n i i i >:;lOO. Anal. (C22HmCltN6) C, €1, S . 9,9'-( 1,4-Piperazinediyldiimino)bis(6-chloro-2-methoxyacridine) (1Xj.--A mixtiire of 9.3 g (0.080 mole) of 1,4-dianiino49.0 g (0.18 mole) of 6,9-dichloro-P-methosyacridine, piperaai~ie,~ and 220 g of phenol was heated oii a ste:tm bath fur 4 hi.. Excess coiicciitrated HC1 was added, aiid the mistiire was diliiied Tvith 1.5 1. of ,\Ie2C0. The resulting precipitate was collected by filtration and washed with 31ed.X. The cariitie hydrochloride was suspended in HZO, excess NaOH was added, and the resiilting base was extracted iiito warm EtOAr. 1-poi1 standing, crystals were deposited. These were collected by filtrat,ion, washed successively wit,h hot 75y0EtOH and Me2C0, arid dried at, 60" in uucuo to give 26.5 g (%yo) of yellow-orange crystals, m p 251 ". :lntd. (C32H&12N602) H, X ; C : calcd, 64.10; foiind, 63.68. 7-Chloro-4-{ [2-(dimethylamino)ethoxy]amino] quinoline Dihydrochloride (XIIa).-A niixt>ure of 9.9 g (0.05 mole) of 4 , i dichloroqiiinoline, 8.9 g (0.05 inole) of 'L-(di~neth~.laniirio)ethoxyamine dihydrochloride,l2 and 23 g uf phenol wis stirred and heated on a steam bath for 4 hr. The resulting brown soliition was cooled and poured into ?vIe&O. The g m i m y product which formed solidified on standing and was dried i n N I C L ( O . Crystallization from i-PrOH afforded 5.9 g (SS(,:; ) of product, mp 213-216" dec. dnal. (CJ~HI~CIN~O.''HC:I.~.~~FII?O) C, H, N, 1120. 7-Chloro-4-( [2-(diethylamino)ethoxy] amino;quinoline (XIIb).
Lincomycin. IX. 7-Thiol and Thioamido Analogs of Lincomycin'
7-Deoxy-7([2)- and -7(S)-thioliiicolnyci11 (IO arid 15) were prepared frunl nietklyl thioliiicosanliliide, it degradation product of lincomycin. Thioamido analogs (18 and 20) were prepared f r o m lincomycin i l i and clindamycin (16), respectively. The i-thiol analogs possessed slight antibacterial activity, while the thioamido analogs were about one-fourth as active as the antibiotic from which they were derived.
l'oteritintion of antibacterial activit,y of lincomycin a variety of chemical modifications.2 Introduction of halogeri at C-7, prefrixbly ill the 7 ( S ) configurat,ion, enhanced t'h(, i i ~ oitru arid 1'11 vivo :mt,ibact,erialactivity in several series of a~ialogs.~ This subst'it'uent,is also primarily responsible for the aiit'imalnrial activit'y encountered in 1'-demethylclindam y c i ~ i s . ~Introduction ~,~ of ot,her substituents at C-7 was therefore undertaken. The preparation arid antibacterial activity of 7-thiol analogs of lincomycin are I ~ O W described. In :iddit,ion to the int~roduct~iori of
(1) was achieved by
(1) Presented in part a t the 11th Medicinal Chemistry Sxmyosium, Quebec. Canada, June 1968. ( 2 ) U. J. hlagerlein, R. D. Birkenmeyer. arid F. liagan, Aittimicrobi,il Agents Chemotherapy-1966, 727 (1967). ( 3 ) (a) D. J. Alagerlein, R. D. Birkenmeger. and F. liagan, J . M e d . L ' h p m . . 10, 355 (1967); ih) R. J. hlagerleinand F.Ragan, ibzd., 12, 780 (lV69). 14) C. Lewis, J . I'orasitol., 6 4 , 169 (1968).
rk: I
@COSH mf
LINCOMYCIN. IX
Sovember 1969
975
CHARTI CI1, RHN
-
Ro@sMe
-
HoQ
-
HJ
Ho@
SMe
OR
OH
PHA NH
"OQSMe
SMe OH
\
OH
4,R=H 5 . R = AC
11
RS Ac -HN+
$I CHsCSHN
-
----t
SMe ' OAc 7,R=H 8, R = AC
6
PHA =
1
CH,CONH
-
Ho@sMe OH
2
4
CH,CONH
'
3
OH
I
'
I
OH
9
1
OH
10
GoCJH;n
I n a previous paper in this series3bsolvolysis of methyl S-acetyl-7-deoxy-7(S)-chlorothiolincosaminide(2) to methyl N-acetylthiolincosaminide (3) mas shown to be ctnchimerically assisted by the amide carbonyl and we suggested that the reaction proceeded through oxazol-
c1-
PHAHN+ Ho@sMe
SMe
I
OAc
-
Ho@
H S T
onium (4). Since the oxygen of the amide carbonyl is postulated to transfer to C-7 with inversion during this reaction, solvolysis of a 7-deoxy-(S)-chlorothioamide should result in the introduction of a 7(R)-thio substituent at C-7. To test this hypothesis (cf. Chart I) methyl 7-deoxy-7(S)-chlorothiolincosaminide tetraacetate ( 5 ) was converted to thioamide 6 in good yield by treatment with P&. Solvolysis in aqueous D M F afforded the predicted product, methyl 7-deoxy-7(R)thiolthiolincosaminide tetraacetate (7). Structural assignment of 7 was made on the basis of elemental analysis and nmr and mass spectral data. Deacylation of 7 to the amino sugar 9 with hydrazine hydrate required milder conditions than employed in the cleavage of lincomycin;5 however, greater difficulty mas encountered in the purification of the product 9. Nore vigorous hydrazinolysis conditions led to the formation of a mixture of products accompanied in part by loss of hydrogen sulfide. I n one attempt to circumvent this problem, thio sugar 7 was oxidized to disulfide 11 which was then treated with hydrazine hydrate. From the resulting mixture a reduction product, whose structure is formulated as 12, was isolated in low yield. ( 5 ) W. Sohroeder, B. Bannister, and 89,2448 (1967).
H. Hoebema, J . Am. Chcm. Soc.,
.'
I
16, R,= R,= H: K,, = C1; N = 0 17, R, = Ac; R?= 0.4~;R ; = H;X = S 18,Rl=Rn=H; R 2 = O H ; X = S 19, R, = Ac; R,= H;R , =Cl: X = S 20. R , = R l = H : R , =
t,ioii 7 by the solvolytic method described above (6 --t 7) \vas t'ried o n it *mal1 scale. Tlc of the solvolytic product of 19 after alkaline hydrolysis indicated t,hc ~ ) r c ; . ; r r i c c of' ~ 10, identified by tlc using chemical visualizaI ion and bioautograph 2's. SarciTia lutea. Antibacterial Activities ---Aiitibact'erial testing data tor the 7-thio and thioamido :riialogs of lincomycin described in this paper are summarized in Table I. The thioamido analogs 18 and 20 possess about 20-25% of the antibacterial pot'ency of t,he parent antibiotics. The 7 ( R ) -and 7(S)-thio analogs 10 and 15 are about lo?(, as act'ive as lincomycin in t,he standard curve S . lutea assay, but surprisingly possessed considerably grcater potency vs. Staphylococcus when administered si1l)cutaneouslg in the mouse protect'ion assay. Broth ( 6 ) 13. J. SIaperlein, K. 13. Birkenmeyer. R . K . Herr. and F. Kagan. J ' A m . C h e m . S o c . , 89, 2459 (1967). 17) R . Guthrie and King, C d i o h y d r u k Re*., 3, 128 (1967).
n.
n.
Sovember 1969
DIURETIC AGEXTS
977
4 : l ) followed by crystallization (H20) yielded 100 mg of 12, nip apparently accompanied by loss of HSS. hiass spectrographic 178-182". Ir, mass spectra, and nmr data indicated identity examination of the crystals showed the expected >I+. This with 12 pi,epared by me1 hod A. product was wed in the nest st,ep where separation of a pnre Thioamidolincomycin Tetraacetate Hydrochloride (17 'Ha).prodrict, proved less vompIic*a1.ctl. 7-Deoxy-7(R)-thiolincomycin(10). --Jlet hyl 7-tleosj-7 ( R ) - Jincomycin tetraacetate (19.5 g), prepared by acylation of lincomycin wit,h Ac$3-C,HiN, was treated with 7 g nf PSIOin aqueous I hinlincosaminide (9) (.5.2 g ) w a l ~lrmted with 4.14 g of trons-4-ndioxane as previously described. Conversion of t,he crude prodI)ropyl-l.-proliIir::~1 o give ti.26 g of crude prodiict. Chroniatoguct tCJ the hydrochloride yielded 24.6 g of 17.IICl which gave 9.0 raphy (CIICI4-lIe0ll,4: 1) gave 5.M g of 10, [ a ] uf123' (HzO), as an amorphoLis solid. The hydrochloride salt also wah not g of 17.IIC1, mp 207-214", from AcCH,-H?O. Recrystallization gave 17.HCI, mp 221-225" (hygroscopic). And. (C2BH43crystalline. Anal. (C,8H8,SZO&) C, 11, iY. Methyl 7-Deoxy-6-deamino-6,7-aziridinothiolincosaminide ClNPOgSP) C, H, N, S, C1. Thioamidolincomycin (IS).-Five grams of 17.HCl was treated (13).--hmiIlo sugar 4 (1 g ) was treated with &CO3 in boiling with 0.5 S 50% aqueous methanolic S a O H a t 26' for 1 hr. The I)MF. Crystallization of the residne from NeOH after distillacrude product (3.5 g) was obtained by extraction with MeCL. tion of the DMF afforded 370 mgof 13, mp 192-198', and 160 mg, Crystallization (aqueous MeOH) afforded 300 mg of 18, mp 221mp 182-189'. Two recrystallizations (1IeOH-AcCH3) gave 226", and b second crop of 530 mg of 18, mp 195-205'. The a h , mp 20:1-220D der, [ L Y ] D f320' (1)lIPo). Only one analytical s,imple was further purified by chromatography (Etspot was noted on tlc and the melting point was unchanged for OA4c-Ac31e-HP0, 8:5:1). Anal. (ClsHz4N20&) C , H, N, S. further recryitallizationr. ;Lnal. (Cc,H,iS04P)C, H, iY. Thioamidoclindamycin Triacetate (19).-1n t,he manner deMethyl 7-Deoxy-7(S)-thiolincosaminide (14).--A suspension cribed above, clindamycin was converted to the triacetate and of 1.5 g of aziridiue 13 i n 30 ml of i-PrOH saturated with H2S a t treat.ed with P&O. After twice chromatographing (CsHiP-Ac0" was heated in a glass bomb on a steam bath for 5 hr. FiltraMe, 2 : l ) 120 mg of 19, mp 178-181", was obtained from 4 g of tion of the cooled reaction mistmureyielded 1.6 g of 14, mp 190t'riacetat'e. A n a l . (C2rH39C1K~0& C, H, N, S. 197", which when recrystallized (Et'OH) melt'ed at 193-198". 7-Deoxy-7(S)-chlorothiamidolincomycin (ZO).-Triacetate 19 .tna/. (C,H,,SO,S,) C, H, X, S. (12.9 mg) was treated wit,h dilute alkali in aqueous AcMe for 25 The pentaaretate (14a), mp 266-268", was prepared by acylamin. Only one spot was noted on tlc moving about \There extion of 14 with AryO-CjH:N in the Iisiial manner. Anal. ((319pected. The solution was acidified and lyophilized. PnrifiraII?,SO,S,) c, H, N, s. tion was not attempted. 7-Deoxy-7(S)-thiolincomycinHydrochloride (15).-In the Solvolysis of 19.--9 solution of 10 mg of 19 in 5070 aqueous manner previously described,Zb 6.8 g of 14 was condensed with 5.2 DMF was heated a t reflux for 17 hr. A more polar product g of trans-4-n-propyl-L-proline to yield 10.2 g of crude product. which reacted very rapidly with Lemieux reagent (characteristic Chromat,ography (CHCI3-MeOH, 4 : l), followed by conversion of thiols) was noted on tlc. The p H --as adjusted t o 10.5 and after t o t,he hydrochloride, afforded 4.35 g of amorphous solid, [.ID 1 hr evaporated in vacuo. The major spot on tlc (CsH12-AcMe, -161" (HyO). d n a l . (C,8H3sC1NaO&) C, H, N. Iiechro2 : l ) (CHC13-MeOH, 1 0 : l ) moved with and was not separated mat.ography of the free ba,qe gave an analyt'ical sample. Anal. from 10 on &in. tlc plates. For visualization Lemieux reagent, (C,,H~,NPO&) C, H, S, S. 1 2 , and bioautograph us. S. lutea were employed. Methyl 6-Deamino-7-deoxythiolincosaminide(12). Method A,-Aziridine 13 (235 mg) was treated with HON07 followed by Acknowledgment.-The authors are indebted t,o Dr. catalyt,ic hydrogenation over 10% Pd-C in MeOH t,o give 60 mg D. J. Mason for in vitro antibacterial t'esting, t'o C. Lewis of 12 (,lcCH1), mp 189-190". Anal. (CgH,,O$) C, H, S. for in vivo assays, and to R . J. Reid for technical Method B.--l)isiilfide 11 (1.4 g ) was heated at reflux for 18 hr assist'ance. n-ith hydrazine hydrate. Chromat,ographjr (CHCI3-MeOH,
The Relationship between Structure, Stereochemistry, and Diuretic Activity in the 2-Amino-a-phenylcyclohexanemethanol Series, a New Class of Diuretic Agents. 111
L. L. SKALETZKY, B. E. GRAHAM, AND J. SZJIUSZKOVICZ Research Laboratories of The Upjohn Company, Kalanaazoo, Michigan
@OOl
Received M a y 29, 1969
A series of lJ3-amino alcohols and lJ3-amino ethers were made for diuretic t'esting. The four diastereoisomers of both the alcohols and methyl et,hers (I, R = H and CH,, respectively) were synthesized and the relationship of stereochemistry to diuretic activity is discussed. 1- [2-(p,aY-Dimethoxybenzyl)cgclohexyl]piperidine (2) was resolved and t,he d enantiomorph (3) was chosen for further pharmacologic evaluation.
I n this paper, a new class of diuretic agents2 illustrated by structure I is reported. Compounds of structure type I have three asymmetric centers and, in the case of both the alcohols and methyl ethers (I, R = H and CH3, respectively), the four possible diastereoisomers or racemates have been synthesized. Stereochemistry is a prime factor in the correlation of structure with biological activity and in the study of
receptor surface^.^ We therefore undertook a detailed study of the relationship between stereochemistry and diuretic activity in this class of compounds.
(1) Paper I : J. Szmuszkovicz and L. L. Skaletzky, J . Org. Chem., S I , 3300 (1967). (2) For a review of diuretic agents: (a) H . J . Hess in "Annual Reports in
I
Medicinal Chemistry, 1967," C. K. Cain, Ed., Academic Press Inc.. New York, K. Y.. 1968, p 62,and previous Annual Reports; (b) G. de Stevens, "Diuretica," Academic Press, Inc., New York, N. Y., 1863.
(3) (a) E . J. Ariens, M o l . Pharmacal., 1, 232 (1964); (b) P. 9. Portoghese, A. A. Mikhail, and H. J. Kupferberg, J . M e d . Chem., 11, 219 (1968), and references noted therein.
