# Patent

## Title
Imidazolyl substituted steroidal and indan-1-one derivatives

## Abstract

The present invention provides a novel series of imidazolyl substituted steroidal and indan-1-one derivatives and salts thereof having the following general structural formulae (A and B)

## Background
This application is a §371 national stage of PCT International Application No. PCT/IB2006/002471, filed Sep. 8, 2006, and claims priority of Indian Patent Application No. 2498/DEL/2005, filed Sep. 15, 2005, the contents of all of which are hereby incorporated by reference into this application.
The present invention relates to Novel series of imidazolyl substituted steroidal and indan-1-one derivatives. The present invention also relates to novel imidazole ring substituted steroidal as non-steroidal aromatase inhibitors. This invention particularly relates to provide the process of preparation of such derivatives and their biological activity as potent aromatase inhibitory compounds.
Aromatase is the physiological enzyme responsible for the conversion of androgens such as androstenedione or testosterone, into estrogens e.g., estrone and estradiol, respectively (Simpson E R et al, Endocrine Reviews, 1994, 15: 342-355). Inhibition of aromatase, a cytochrome P-450 enzyme, has become a strategy of choice to interfere with normal or pathological estrogen-induced or estrogen-dependent biological processes such as female sexual differentiation, ovulation, implantation, breast and endometrial cell proliferation as well as regulation of spermatogenesis or prostate cell proliferation of male or of non-reproductive functions such as bone formation or Immune T cell and cytokine balance (Simpson E R et al., Recent progress in Hormone research, 1997, 52: 185-213) and also in the treatment of disseminated estrogen dependent breast cancer over the last three decades (Tomera J. F., Drugs of Today, 1994, 30, 565-574; Howell A., Dowsett M., BMJ. 1997).
A number of steroidal and non-steroidal compounds affecting estrogen biosynthesis through the inhibition of aromatase are presently in the market and others are in various stages of clinical trials for the treatment of breast cancer. (Njar, V. C. O; Brodie, A. M. H, Drugs, 1999, 58, 233-255; Seralini G E, Moslemi, S. Mol. Cell. Endocrinol, 2001, 178, 117-131).
The steroidal inhibitors are generally more specific, but have the potential to induce unwanted agonist effects. In contrast, non-steroidal inhibitors may lack specificity because they have the potential to block several P450 mediated steroidal conversions but are less likely to exhibit unwanted agonist effects of steroids.
Steroidal inhibitors such as 4-hydroxyandrostenedione (formestane) (Carlini, P. et al., Ann. Oncol., 2001, 12, 1539-1543) and atamestane (Traina, T A. et al., Curr. Opin. Invest. Drugs., 2004, 605-610) compete with the endogenous substrates, androstenedione and testosterone, for the active site of the enzyme, where they act as false substrates and are processed to intermediates that bind irreversibly to the active site, causing irreversible enzyme inhibition.




Nonsteroidal inhibitors also compete with the endogenous substrates for access to the active site, where they then form a coordinate bond to the heme iron atom. Therefore, they effectively exclude both the natural substrate) and oxygen from the enzyme.
A large number of azole derivatives are traditionally known as antifungal agents. Some of the imidazole and triazole derivatives have already been described as inhibitors of enzyme aromatase. Generally the imidazolyl and triazole groups are generally associated with aromatic rings as in letrozole (Buzdar A et al., Proc. Am. Soc. Clin. Oncol. 1996; Lamb H M and Adkins J C, Drugs, 1998, 56: 1125-1140) and anastrozole (Wiseman L R and Adkins J C, Drugs Aging, 1998, 13, 321-332) or linked via a methylene group to a benzotriazole are described in EP-A-293-978. Fadrozole, an imidazole derivative, is a potent competitive inhibitor of aromatase both in vitro and in vivo and is more selective than aminoglutethimide.




Hartmann et al (J. Med. Chem., 1994, 37, 1275-1281) had also described some pyridyl substituted indanones and tetralones as potent and selective non-steroidal anti-aromatase agents, when compared with standard drug aminoglutethimide.




Further, their hydroxy and methoxy derivatives at 5 and 6 positions have also shown 150-200 times more activity as compared to aminoglutethimide.
Accordingly, the main object of this invention is to provide certain new imidazolyl substituted steroidal and indanone derivatives. Another object is to provide a process for preparation of such newer steroidal as well as non-steroidal aromatase inhibitors.
Further object of the invention is to provide the imidazolyl substituted steroidal and indanone derivatives which showed better relative potency as compared to standard drug aminoglutethimide.
Accordingly, the present invention provides a novel series of imidazolyl substituted steroidal and indan-1-one derivatives and salts thereof having the following general structural formulae (A and B)




wherein R is




R1, R2 and R3 independently represent H, or —OCH3 or




R4 and R5 independently represent —OH, or ═O or —OCOCH3 groups
R6 independently represents H or




moieties.

n=2 or 3, A double bond is either present between 4 and 5 or 5 and 6 positions of the steroidal nucleus in case of steroidal derivatives.
In an embodiment of the invention a novel series of imidazolyl substituted steroidal derivatives of formula A, wherein the 16-substituted-androst-5-ene steroidal derivatives having general formula:




In another embodiment of the invention a novel series of imidazolyl substituted steroidal derivatives of formula A, wherein the 16-substituted-androst-4-ene-3,17-dione is having general formula:




In yet another embodiment of the invention a novel series of imidazolyl substituted steroidal derivatives of formula A, wherein the 7-substituted-androst-5-ene steroidal derivatives having general formula:




In a further embodiment of the invention a novel series of imidazolyl substituted steroidal derivatives of formula A, wherein the structure of 16-substituted steroidal compound is




In a further embodiment of the invention a novel series of imidazolyl substituted steroidal derivatives of formula A, wherein the representative compounds of imidazolyl substituted steroidal derivatives are:

    16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-17-oxo-5-androst-en-3β-ol (12) (DPJ-RG-1151)
    16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-17-oxo-5-androsten-3β-yl acetate (13) (DPJ-RG-1196)
    16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-5-androstene-3β,17β-diol (14) (DPJ-RG-1219)
    16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-5-androstene-3β,17β-diol diacetate (15) (DPJ-RG-1227)
    16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-4-androstene-3,17-dione (16) (DPJ-RG-1177)
    16-[3-{3-(imidazol-1-yl)propoxy}benzylidene]-17-oxo-5-andro-sten-3β-ol (18) (DPJ-RG-1307)
    16-[3-{3-(imidazol-1-yl)propoxy}benzylidene]-17-oxo-5-andro-sten-3β-yl acetate (19) (DPJ-RG-1309)
    16-[3-{3-(imidazol-1-yl)propoxy}benzylidene]-5-androstene-3β,17β-diol (20) (DPJ-RG-1310)
    16-[3-{3-(imidazol-1-yl)propoxy}benzylidene]-5-androstene-3β,17β-diol diacetate (21) (DPJ-RG-1311)
    16-[3-{3-(imidazol-1-yl)propoxy}benzylidene]-4-androstene-3,17-dione (22) (DPJ-RG-1308)
    7-[O-{3-(Imidazol-1-yl)propyl}oximino]-5-androstene-3β,17β-diol (25) (DPJ-RG-1223)
    16β-(Imidazol-1-yl)-17-oxo-5-androsten-3β-ol (27) (DPJ-RG-1240)
    16β-(Imidazol-1-yl)-5-androstene-3β,17β-diol (28) (DPJ-RG-1317)
    16β-(Imidazol-1-yl)-5-androstene-3β,17β-diol diacetate (29) (DPJ-RG-1318)
    16β-(Imidazol-1-yl)-4-androstene-3,17-dione (30) (DPJ-RG-1241)


In another embodiment of the invention a novel series of imidazolyl substituted indan-1-one derivatives of formula B, wherein the indan-1-one derivatives have general formula as:




In another embodiment of the invention a novel series of imidazolyl substituted indan-1-one derivatives wherein, representative compounds of indan-1-one derivatives are:

    2-[3-(3-Imidazol-1-yl-propoxy)-benzylidene]-indan-1-one (2) (DPJ-RG-1088)
    2-[2-(3-Imidazol-1-yl-propoxy)-benzylidene]-indan-1-one (4) (DPJ-1055)
    2-[3-(3-Imidazol-1-yl propoxy)-methoxybenzylidene)indan-1-one (6) (DPJ-RG-1090)
    2-[4-(3-Imidazol-1-yl propoxy)-3-methoxybenzylidene)indan-1-one (8) (RG-DPJ-195)
    2-[3-(2-Imidazol-1-yl ethoxy)-benzylidene]indan-1-one (10) (RG-DPJ-325)
    2-(1H-1-imidazolyl)-1-indanone (32)


In another embodiment of the invention a novel series of imidazolyl substituted indan-1-one derivatives wherein the salt may be selected from any pharmaceutically acceptable salt.
In another embodiment of the invention a novel Use of the compounds wherein the compounds having general formula A and B shows aromatase inhibitory activity.
In another embodiment of the invention the novel compounds of the series wherein the IC50 values for the aromatase inhibitory activity of the potent compounds 2, 4, 6, 8, 14, 16, 20, 22, 24, 25, 27 and 30 is ranging between 0.18-12 μM with relative potency of these compounds ranging from 2.5 to 165.3 times as compared to standard drug aminoglutethimide.
The novel compounds wherein the compounds are useful as anticancer agents. Accordingly, the present invention provides a process for preparation of 16-substituted-androstene compounds of formula A comprising; base catalysed aldol condensation of DHA with chloroalkylated aldehydes of formula C at room temperature to afford halogenated aldol adducts and followed by thermal fusion of these chloroalkoxybenzylidene derivatives of DHA with imidazole at a temperature ranging between 130-140° C., to afford imidazolyl substituted aldol products, converting the said aldol products obtained into corresponding 5-androstene-3,17-diacetate compounds or oxidizing to 4-androstene-2,17-dione compounds or to 5-androstene-3-aceoxy-17-one or to 5-androstene-3,17-diol compounds by known methods.
In another embodiment of the invention a novel process wherein 3-acetoxy-17-one-5-androstene steroid is prepared from 3-hydroxy precursor by treatment with acetic anhydride and dry pyridine in steam bath for about 2 hrs.
In another embodiment of the invention a novel process wherein androst-5-en-3,1-diol was formed by sodium borohydride reduction of 3-hydroxy-17-one-5-androstene steroid at room temperature which on treatment with acetic anhydride and pyridine in steam bath for about 2 hrs gave 3,17-diacetoxy-5-androstene aldol products.
In another embodiment of the invention a novel process wherein the preparation of 4-androstene-3,17-dione steroidal aldol products were prepared by Oppenauer oxidation of 5-androstene-3-hydroxy-17-one aldol products. Accordingly the present invention provides a novel process for the preparation of 7-substituted-5-androstene steroidal derivatives possessing Imidazole ring of formula A, comprising: treating 7-hydroximino-5-androstene derivative, prepared by known methods, with 1-bromo-3-chloropropane and subsequent thermal fusion with imidazole and followed by its hydrolysis.
In an embodiment of the invention a novel process for the preparation of 16-Imidazolylandrostene steroids comprising; brominating DHA by refluxing with cupric bromide in methanol and benzene to give 16-bromo derivatives and their subsequent thermal fusion with powdered imidazole at a temperature ranging between 130-140° C.
Accordingly the present invention provides a Process for preparation of 2-aminoalkoxybenzylidene-1-indanone of formula B, comprising; base catalysed aldol condensation of 1-indanone with chloroalkylated aldehydes of formula C at room temperature to afford halogenated aldol adducts followed by thermal fusion of chloroalkoxybenzylidene derivatives of 1-indanone with imidazole at a temperature ranging between 80-90° C., to afford imidazolyl substituted aldol products of formula B.
In an embodiment of the invention a process for the preparation of 2-imidazolyl-1-indanone wherein the said compound is prepared by thermal fusion of 2-bromo-1-indanone with powdered imidazole at a temperature ranging between 80-90° C.
In the embodiment of the present invention, the aromatase inhibitory activity of these compounds is monitored by measuring the 3H2O formed from [1β,2β-3H] testosterone during aromatization. These compounds produced a concentration dependent inhibition of aromatase, which in many cases is significantly higher than that of standard drug aminoglutethimide.


DETAILED DESCRIPTION
The compounds which comprise in the present invention have the following general structural formulae (A and B)




    
    
        Wherein R independently represents
    
    






    
    
        R1, R2 and R3 independently represent H, or —OCH3 or
    
    






    
    
        R4 and R5 independently represent —OH, or ═O or —OCOCH3 groups
        R6 independently represents H or
    
    






    
    
         moieties.
        n=2 or 3
        A double bond is either present between 4 and 5 or 5 and 6 positions of the steroidal nucleus in case of steroid derivatives.
    
    


In preferred compounds of the invention, of outstanding interest are: 2-[3-(3-Imidazol-1-yl-propoxy)-benzylidene]-indan-1-one (2) (DPJ-RG-1088); 2-[2-(3-Imidazol-1-yl-propoxy)-benzylidine]-indan-1-one (4) (DPJ-1055); 2-[3-(3-Imidazol-1-yl propoxy)-4-methoxybenzylidene)indan-1-one (6) (DPJ-RG-1090); 2-[4-(3-Imidazol-1-yl propoxy)-3-methoxybenzylidene)indan-1-one (8) (RG-DPJ-195); 2-[3-(2-Imidazol-1-yl ethoxy)-benzylidine]indan-1-one (10) (RG-DPJ-325); 2-(1H-1-imidazolyl)-1-indanone (32); 16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxy-benzylidene]-17-oxo-5-androsten-3β-ol (12) (DPJ-RG-1151); 16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxy-benzylidene]-17-oxo-5-androsten-3β-yl acetate (13) (DPJ-RG-1196); 16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-5-androstene-3β,17β-diol (14) (DPJ-RG-1219); 16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-5-androstene-3β,17β-diol diacetate (15) (DPJ-RG-1227); 16-[4-{3-(Imidazol-1-yl)propoxy}3-methoxybenzylidene]-4-androstene-3,1-dione (16) (DPJ-RG-1177); 16-[3-{3-(imidazol-1-yl)propoxy}benzylidene]-17-oxo-5-androsten-3β-ol (18) (DPJ-RG-1307); 16-[3-{3-(imidazol-1-yl)propoxy}benzylidene]-17-oxo-5-andro-sten-3β-yl acetate (19) (DPJ-RG-1309); 16-[3-{3-(imidazol-1-yl)propoxy}benzylidene]-5-androstene-3β, 17β-diol (20) (DPJ-RG-1310); 16-[3-{3-(imidazol-1-yl)propoxy}benzylidene]-5-androstene-3β,17β-diol diacetate (21) (DPJ-RG-1311); 16-[3-{3-(imidazol-1-yl)propoxy}benzylidene]-4-androstene-3,17-dione (22) (DPJ-RG-1308); 7-[O-{3-(Imidazol-1-yl)propyl}oximino]-5-androstene-3β,17β-diol (25) (DPJ-RG-1223); 16β-(Imidazol-1-yl)-17-oxo-5-androsten-3β-ol (27) (DPJ-RG-1240); 16β-(Imidazol-1-yl)-5-androstene-3β,17β-diol (28) (DPJ-RG-1317); 16β-(Imidazol-1-yl)-5-androstene-3β,17β-diol diacetate (29) (DPJ-RG-1318); 16β-(Imidazol-1-yl)-4-androstene-3,17-dione (30) (DPJ-RG-1241)
The steroidal as well as non-steroidal compounds of general formulae (A) and (B) (wherein R is




and R1, R2, R3, R4, R5 and R6 are as herein before defined) which comprise the present invention have been prepared by the aldol condensation of some alkylated aldehydes with steroidal or □ndanones nucleus. These aldol products were then subjected to fusion with imidazole to get the desired products.

The alkylated aldehydes of general formula © (where R1, R2 and R3 independently represent H or —OCH3 or —O(CH2)nCl, n=2 or 3) were prepared by treating the corresponding aldehydes; salicylaldehyde, 3-hydroxybenzaldehyde, 4-hydroxybenzaldehyde, isovanillin or vanillin with either 3-bromo-1-chloropropane or 2-bromo-1-chloroethane in the presence of anhydrous potassium carbonate.




These aldehydes of general formula © were then subjected to aldol condensation [Nielsen, A. T.; Houlihan, W. J. The aldol Condensation. In Organic Reactions, 16th ed. (John Wiley: New York) 1968; 1-438] with dehydroepiandrosterone (DHA) or with indan-1-one to give various aldol products, and were subsequently thermally fused with imidazole, which has been described as a pharmacologically active heterocyclic ring with potential for aromatase inhibition in literature [Adje, N. et al 1-N-Phenylamino-1H-imidazole derivatives as aromatase inhibitors and pharmaceutical compositions containing them, (Laboratoire Thermax) U.S. Pat. No. 6,737,433 B1, 2004], to develop new derivatives of interest for aromatase inhibition of general formulae (A) and (B) wherein R is




and R1, R2 and R3 are herein as before described and in case of steroidal derivatives, R4=—OH, R5 represents ═O, R6=H and a double bond is present between 5 and 6 positions.

According to a feature of present invention, the steroidal derivatives of general formula (A) wherein R4 independently represents —OCOCH3 group and R5 independently represent ═O group, R6=H and a double bond is present between 5 and 6 positions were in turn prepared by acetylation of 17-keto-3-hydroxy DHA aldol product with acetic anhydride in dry pyridine.
According to a feature of the present invention the steroidal aldol products of general formula (A) wherein both R4 and R5 independently represents —OH group, were inturn prepared by reducing the 17-keto-3-hydroxy DHA aldol product with sodium borohydride in methanol.
The derivatives of general formula (A) wherein both R4 and R5 independently represent —OCOCH3 group was prepared by direct acetylation of previously described 3,17-dihydroxy aldol products with acetic anhydride in dry pyridine.
According to a feature of present invention, the derivatives of general formula (A) wherein both R4 and R5 represent ═O group and double bond is present between 4 and 5 positions of steroid nucleus were prepared by Oppenauer oxidation [Eastham, J. F.; Teranishi, R. In Organic Synthesis, Cairns, T. L.; Ed; (John Wiley: New York) 1955; 39] of 17-keto-3-hydroxy DHA aldol product in aluminium isopropoxide-cyclohexanone-toluene system.
According to another feature of the present invention the derivatives of the general formula (A) wherein R independently represents hydrogen and R6 independently represent




groups and R4=R5=—OCOCH3 or —OH, have been prepared by allylic oxidation of diacetyl derivatives of dehydroepiandrosterone acetate using t-butyl chromate reagent in CCl4 by the method described in the literature (Heusler, K. et al, Helv. Chim. Acta, 1952, 35, 284-294) to form a product wherein R6 represents ═O. This was further treated with hydroxylamine hydrochloride to form a derivative with R6 representing ═NOH (Singh H et al., Indian J. Chem., 1969, 1084-1087), and then with 1-bromo-3-chloropropane and subsequently thermally fused with imidazole and hydrolysed.

The derivatives of general formulae (A) and (B) wherein R is represented by




have been prepared by bromination of DHA or indan1-one using cupric bromide by a method previously described in literature (Glazier E R, J. Org. Chem., 1962, 2937-2938) and subsequent thermal fusion with powdered imidazole. Further modifications at R4 and R5 have been carried out as previously described in case of steroidal derivatives.

Biological Activity

The in vitro aromatase inhibitory activity was carried out using human placental microsomes and [1β,2β-3H] testosterone by a method previously described in literature (Graves, P E, Et al, Endocrinology, 1979, 52-57). The imidazolyl substituted □ndanones derivatives 2, 4, 6 and 8 displayed good aromatase inhibitory activity with compound 8 (RG-DPJ-195) exhibiting IC50=0.55 μM with 50 times greater activity in comparison to aminoglutethimide. These hybrid molecules of □ndanones and imidazole resulted in enhanced activity and may have good potential for the treatment of breast cancer. The imidazolyl substituted steroidal derivatives 14, 16, 20, 22, 24, 25, 27 and 30 exhibited significant aromatase inhibitory activity. Compound 30 (DPJ-RG-1241) was the most valuable compound of the series with IC50=0.18 μM and relative potency 165 in comparison to aminoglutethimide.
Following examples are given by way of illustration and should not construed to limit the scope of the invention.
EXAMPLE-1
a) 3-(3-Chloropropoxy)benzaldehyde
To a stirred and refluxing suspension of 3-hydroxybenzaldehyde (1.0 g, 8.19 mmol) and potassium carbonate (2 g) in ethyl methyl ketone, 1-bromo-3-chloropropane was added and reaction mixture was allowed to reflux with stirring on oil bath for 6 h. The completion of reaction was monitored by TLC. The slurry was filtered and solvent removed under reduced pressure to obtain an oily residue, which was used as such for further reaction [Naruto S, et al., J. Med. Chem. 1982, 25, 1240-1245].
2-(3-chloropropoxy)benzaldehyde, 3-(3-chloropropoxy)-4-methoxybenzaldehyde and 4-(3-chloropropoxy)-3-methoxybenzaldehyde were prepared using corresponding aldehydes such as salicylaldehyde, isovanillin and vanillin. 3-(2-chloroethoxy)-benzaldehyde was prepared by treating 3-hydroxybenzaldehyde with 1-bromo-2-chloroethane using the similar method described above.
b) 2-[3-(3-Chloropropoxy)benzylidene]-indan-1-one (1) (DPJ-RG-1054)
A solution of indan-1-one (0.4 g, 3.03 mmol) and sodium hydroxide (0.6 g, 15 mmol) in methanol (25 ml) was added to the solution of 3-(3-chloropropoxy)benzaldehyde in methanol (10 ml). The resulting solution was shaken for 30 min and allowed to stand at room temperature with intermittent shaking. The completion of reaction was monitored by TLC. After the completion, excess of methanol was removed; crushed ice was added and allowed to stand overnight. The precipitates obtained were filtered, washed thoroughly with distilled water and dried. The crude product was crystallized from a mixture of acetone and petroleum ether to yield white colored crystals of 2-[3-(3-chloropropoxy)benzylidene]-indan-1-one (1) (0.21 g, 33.5%), mp 73-75° C.
IR (KBr): 2960, 1690, 1590, 1430, 1230 and 740 cm−1.
1H-NMR (CDCl3): δ 2.28 (p, 2H, —OCH2CH2—), 3.78 (t, 2H, —CH2Cl), 4.05 (s, 2H, 3-CH2 indanone), 4.18 (t, 2H, —OCH2—), 6.96 (dd, 1H, Jo=8.85 Hz, Jm=2.43 Hz, aromatic), 7.42 (m, 7H, vinylic-H and aromatic) and 7.91 ppm (d, 1H, Jo=7.59 Hz, 7-H □ndanones).
Calcd. For C19H17O2Cl: C, 72.95; H, 5.47. Found: C, 72.56; H, 5.05.
2-[2-(3-CHLOROPROPOXY)BENZYLIDENE]-INDAN-1-ONE (3) was prepared using 2-(3-chloropropoxy)benzaldehyde following the method described for 1
2-[3-(3-CHLOROPROPOXY)-4-METHOXYBENZYLIDENE]-INDAN-1-ONE (5) was prepared using 3-(3-chloropropoxy) 4-methoxy benzaldehyde following the method described for 1
2-[4-(3-CHLOROPROPOXY)-3-METHOXYBENZYLIDENE]-INDAN-1-ONE (7) (DPJ-1039) was prepared using 4-(3-chloropropoxy)-3-methoxybenzaldehyde following the method described for 1
2-[3-(2-CHLOROETHOXY)BENZYLIDINE]INDAN-1-ONE (9) (RG-DPJ-324) was prepared using 3-(2-chloroethoxy)benzaldehyde following the method described for 1
c) 2-[3-(3-Imidazol-1-yl-propoxy)-benzylidene]-indan-1-one (2) (DPJ-RG-1088)
The compound 2-[3-(3-chloropropoxy)-benzylidene]-indan-1-one (1) (1 g) was triturated with imidazole (1.5 g) and heated at 80-85° C. with stirring for 1 h. The reaction mixture was cooled and distilled water was added to it. Water was then decanted and the solid mass was allowed to dry which was then crystallized from solvent ether to give the compound 2 (0.51 g, 46.59%), mp 48-50° C.
Anal:
IR (KBr): 2940, 1690, 1610, 1240, 1085 and 740 cm−1.
1H-NMR (CDCl3): δ 2.27 (p, 2H, —OCH2CH2—), 3.96 (t, 2H, —CH2N), 4.03 (s, 2H, 3-CH2 indanone), 4.22 (t, 2H, —OCH2—), 6.94 (m, 2H, aromatic and imidazolyl), 7.07 (s, 1H, aromatic), 7.16 (s, 1H, aromatic), 7.35 (m, 3H, two aromatic and one imidazolyl), 7.50 (s, 1H, imidazolyl), 7.58 (m, 3H, aromatic (□ndanones) and vinylic-H) and 7.92 ppm (d, 1H, Jo=7.53 Hz, 7-CH □ndanones)
Calcd. For C22H20O2N2: C, 76.72; H, 5.85; N, 8.16. Found: C, 76.68; H, 5.86; N, 8.10.
2-[2-(3-IMIDAZOL-1-YL-PROPOXY)-BENZYLIDINE]-INDAN-1-ONE (4) (DPJ-1055) was prepared by fusing compound 3 with imidazole following the method described for 2
2-[3-(3-IMIDAZOL-1 YL PROPOXY)-4-METHOXYBENZYLIDENE)INDAN-1-ONE (6) (DPJ-RG-1090) was prepared by fusing compound 5 with imidazole following the method described for 2
2-[4-(3-IMIDAZOL-1YL PROPOXY)-3-METHOXYBENZYLIDENE)INDAN-1-ONE (8) (RG-DPJ-195) was prepared by fusing compound 7 with imidazole following the method described for 2
2-[3-(2-IMIDAZOL-1YL ETHOXY)-BENZYLIDINE]INDAN-1-ONE (10) (RG-DPJ-325) was prepared by fusing compound 9 with imidazole following the method described for 2
EXAMPLE-2
a) 16-[4-(3-Chloropropoxy)-3-methoxybenzylidene]-17-oxo-5-androsten-3β-ol (11) (DPJ-RG-1150)
A mixture of dehydroepiandrosterone (0.75 g, 2.60 mmol), above obtained oily residue of 4-(3-chloropropoxy)-3-methoxybenzaldehyde and sodium hydroxide (1 g) in methanol (10 Ml) was stirred at room temperature for 2 h and the completion of reaction was monitored by TLC. The product was precipitated by addition of cold water and the precipitate obtained was filtered, washed with water, dried and crystallized from methanol to yield 16-[4-(3-chloropropoxy)-3-methoxybenzylidene]-17-oxo-5-androsten-3β-ol (11) (1 g, 76.96%), mp 210-212° C.
Spectral and Elemental Analyses:
Uvmax (MeOH): 243.0 nm (log ε 4.02) and 331.0 nm (log ε 4.36).
FTIRνmax (KBr): 3220.0, 2922.9, 2829.3, 1709.1, 1623.3, 1593.5, 1515.8, 1447.7, 1325.7, 1260.0, 1140.6, 1094.1, 1057.5, 1023.7, 916.4 and 806.0 cm−1.
1H NMR (CDCl3): δ 0.98 (s, 3H, 18-CH3), 1.08 (s, 3H, 19-CH3), 2.31 (m, 2H, —OCH2CH2CH2Cl), 3.53 (m, 1H, 3α-H), 3.77 (t, 2H, —CH2Cl), 3.89 (s, 3H, —OCH3), 4.21 (t, 2H, —OCH2—), 5.40 (d, 1H, 6-CH), 6.95 (d, 1H, Jo=8.27 Hz, 5-CH, aromatic), 7.06 (d, 1H, Jm=1.73 Hz, 2-CH, aromatic), 7.16 (dd, 1H, Jm=1.73 Hz, JO=8.45 Hz, 6-CH, aromatic) and 7.38 ppm (s, 1H, vinylic-H, 16-arylidene).
Calcd. For C30H39O4Cl: C, 72.20; H, 7.88. Found: C, 72.40; H, 8.02.
b) 16-[3-(3-CHLOROPROPOXY)BENZYLIDENE]-17-OXO-5-ANDROSTEN-3β-OL (17) (DPJ-RG-1306) was Prepared Using 3-(3-chloropropoxy)benzaldehyde Following the Method Described for 11
c) 16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-17-oxo-5-androst-en-3β-ol (12) (DPJ-RG-1151)
A mixture of 16-[4-(3-chloropropoxy)-3-methoxybenzylidene]-17-oxo-5-androsten-3β-ol (11) (0.5 g, 1 mmol) and powdered imidazole (0.75 g) was fused at 110-120° C. for 5 h. The completion of reaction was monitored by TLC. The reaction was quenched with cold water and solid obtained was filtered, washed with water, dried and crystallized from ethyl acetate to yield 12 (0.38 g, 71.47%), mp 199-201° C.
Spectral and Elemental Analyses:
UVmax (MeOH): 243.4 nm (log ε 4.04) and 330.8 (log ε 4.39).
FTIRνmax (KBr): 3215.3, 2934.3, 1700.9, 1593.4, 1514.3, 1463.0, 1329.9, 1260.4, 1143.7, 1066.5, 917.2 and 832.0 cm−1.
1H NMR (CDCl3): δ 0.98 (s, 3H, 18-CH3), 1.07 (s, 3H, 19-CH3), 2.30 (m, 2H, —OCH2CH2CH2N<), 3.54 (m, 1H, 3α-H), 3.91 (s, 3H, —OCH3), 3.97 (t, 2H, —CH2N<), 4.23 (t, 2H, —OCH2—), 5.39 (d, 1H, 6-CH), 6.85 (d, 1H, Jo=8.33 Hz, 5-CH, aromatic), 6.93 (s, 1H, 5-CH, imidazole), 7.06 (d, 2H, 2-CH, aromatic and 4-CH, imidazole), 7.13 (d, 1H, Jo=8.26 Hz, 6-CH, aromatic), 7.38 (s, 1H, vinylic-H, 16-arylidene) and 7.48 ppm (s, 1H, 2-CH, imidazole).
Calcd. For C33H42N2O4: C, 74.68; H, 7.98; N, 5.28. Found: C, 74.62; H, 7.82; N, 5.31.
16-[3-{3-(IMIDAZOL-1-YL)PROPOXY}BENZYLIDENE]-17-OXO-5-ANDRO-STEN-3β-OL (18) (DPJ-RG-1307) was prepared by fusing compound 17 with imidazole following the method described for 12
d) 16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-17-oxo-5-androsten-3β-yl Acetate (13) (DPJ-RG-1196)
A mixture of 16-[4-{3-(imidazol-1-yl)propoxy}-3-methoxybenzylidene]-17-oxo-5-androsten-3β-ol (12) (0.5 g, 0.94 mmol), acetic anhydride (1 Ml) and dry pyridine (2 Ml) was heated in a steam bath for 2 h. The reaction contents were then poured into cold water and basified with liquid ammonia. The precipitate obtained was filtered, washed with water, dried and crystallized from n-hexane to afford 13 (0.27 g, 50.04%), mp 109-111° C.
Spectral and Elemental Analyses:
UVmax (MeOH): 249.0 nm (log ε 4.06) and 331.0 nm (log ε 4.30).
FTIRνmax (KBr): 2942.1, 1729.7, 1628.3, 1596.2, 1513.2, 1465.7, 1371.4, 1325.6, 1248.5, 1139.4, 1095.7, 1029.7, 915.1 and 812.7 cm−1.
1H NMR (CDCl3): δ 0.98 (s, 3H, 18-CH3), 1.09 (s, 3H, 19-CH3), 2.04 (s, 3H, —OCOCH3), 2.29 (m, 2H, —OCH2CH2CH2N<), 3.91 (s, 3H, —OCH3), 3.98 (t, 2H, —CH2N<), 4.23 (t, 2H, —OCH2—), 4.61 (m, 1H, 3α-H), 5.42 (d, 1H, 6-CH), 6.85 (d, 1H, Jo=8.48 Hz, 5-CH, aromatic), 6.93 (s, 1H, 5-CH, imidazole), 7.07 (m, 2H, 2-CH, aromatic and 4-CH, imidazole), 7.12 (dd, 1H, Jm=1.44 Hz, Jo=8.26 Hz, 6-CH, aromatic), 7.38 (s, 1H, vinylic-H, 16-arylidene) and 7.50 ppm (s, 1H, 2-CH, imidazole).
Calcd. For C35H44N2O5: C, 73.40; H, 7.74; N, 4.89. Found: C, 73.52; H, 7.95; N, 5.01.
16-[3-{3-(IMIDAZOL-1-YL)PROPOXY}BENZYLIDENE]-17-OXO-5-ANDRO-STEN-3β-YL ACETATE (19) (DPJ-RG-1309) was prepared by acetylation of 18 using the method described for compound 13
e) 16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-5-androstene-3β,1β-diol (14) (DPJ-RG-1219)
To a stirred suspension of 16-[4-{3-(imidazol-1-yl)propoxy}-3-methoxy benzylidene]-17-oxo-5-androsten-3β-ol (12) (1 g, 1.88 mmol) in methanol (100 Ml) at room temperature, sodium borohydride (1.5 g) was added in small fractions over a period of 2 h. The reaction mixture was further stirred for 6 h. Solvent was removed under reduced pressure and iced water was added to it. The precipitate obtained was filtered, washed with water, dried and crystallized from methanol to yield the product 14 (0.85 g, 84.68%), mp 197-198° C.
Spectral and Elemental Analyses:
UVmax (MeOH): 263.6 nm (log ε 4.02).
FTIRνmax (KBr): 3235.9, 2928.7, 1599.3, 1514.0, 1463.4, 1410.2, 1323.1, 1258.8, 1231.7, 1167.5, 1140.4, 1079.6, 1052.1, 949.1, 915.6 and 798.0 cm−1.
1H NMR (CDCl3): δ 0.72 (s, 3H, 18-CH3), 1.05 (s, 3H, 19-CH3), 2.25 (m, 2H, —OCH2CH2CH2N<), 3.53 (m, 1H, 3α-H), 3.89 (s, 3H, —OCH3), 3.95 (t, 2H, —CH2N<), 4.06 (s, 1H, 17α-H), 4.23 (t, 2H, —OCH2—), 5.38 (d, 1H, 6-CH), 6.45 (s, 1H, vinylic-H, 16-arylidene), 6.81 (m, 1H, 5-CH, aromatic), 6.93 (m, 3H, 2-CH, 6-CH, aromatic and 5-CH, imidazole), 7.07 (s, 1H, 4-CH, imidazole) and 7.54 ppm (s, 1H, 2-CH, imidazole).
Calcd. For C33H44N2O4: C, 74.40; H, 8.33; N, 5.26. Found: C, 74.27; H, 8.39; N, 5.39.
16-[3-{3-(IMIDAZOL-1-YL)PROPOXY}BENZYLIDENE]-5-ANDROSTENE-3β,17β-DIOL (20) (DPJ-RG-1310) was prepared by sodium borohydride reduction of 18 using a method as described for 14
f) 16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-5-androstene-3β,1β-diol Diacetate (15) (DPJ-RG-1227)
A mixture of 16-[4-{3-(imidazol-1-yl)propoxy}-3-methoxybenzylidene]-5-androstene-3β,17β-diol (14), 0.5 g, 0.94 mmol), acetic anhydride (1 Ml) and dry pyridine (2 Ml) was heated in a steam bath for 2 h. Contents of the reaction mixture were poured into cold water and basified with liquid ammonia. The precipitate obtained was collected by filtration, washed with water, dried and crystallized from n-hexane to furnish the product 15 (0.38 g, 65.64%), mp 163-165° C.
Spectral and Elemental Analyses:
UVmax (MeOH): 263.8 nm (log ε 4.28).
FTIRνmax (KBr): 2938.9, 1733.7, 1595.9, 1512.1, 1443.3, 1371.1, 1239.5, 1141.3, 1034.2 and 804.5 cm−1.
1H NMR (CDCl3): δ 0.80 (s, 3H, 18-CH3), 1.05 (s, 3H, 19-CH3), 2.04 (s, 3H, 3β-OCOCH3), 2.22 (s, 3H, 17β-OCOCH3), 2.26 (m, 2H, —OCH2CH2CH2N<), 3.89 (s, 3H, —OCH3), 3.95 (t, 2H, —CH2N<), 4.23 (t, 2H, —OCH2—), 4.61 (m, 1H, 3α-H), 5.37 (s, 1H, 17α-H), 5.40 (d, 1H, 6-CH), 6.15 (d, 1H, vinylic-H, 16-arylidene), 6.80 (d, 1H, Jo=8.08 Hz, 5-CH, aromatic), 6.89 (s, 2H, 2-CH, aromatic and 5-CH, imidazole), 6.93 (dd, 1H, Jm=1.78 Hz, Jo=8.86 Hz, 6-CH, aromatic), 7.06 (s, 1H, 4-CH, imidazole) and 7.54 ppm (s, 1H, 2-CH, imidazole).
Calcd. For C37H48N2O6: C, 72.05; H, 7.84; N, 4.54. Found: C, 72.14; H, 7.72; N, 4.66.
16-[3-{3-(IMIDAZOL-1-YL)PROPOXY}BENZYLIDENE]-5-ANDROSTENE-3β,17β-DIOL DIACETATE (21) (DPJ-RG-1311) was prepared by acetylation of 20 by a method described for 15
g) 16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-4-androstene-3,1-dione (16) (DPJ-RG-1177)
16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-17-oxo-5-androsten-3β-ol (12) (1 g, 1.88 mmol) was dissolved in a mixture of cyclohexanone (10 Ml) and dry toluene (150 Ml). Traces of moisture were removed by azeotropic distillation. The distillation was continued at a slow rate while adding a solution of aluminium isopropoxide (1 g) in dry toluene (15 Ml) drop wise. The reaction mixture was refluxed for 4 h. The slurry was filtered and the residue was washed thoroughly with dry toluene. The combined filtrate and the washings were steam distilled until the removal of organic solvents was affected. The crystalline solid was filtered, washed with water, dried and treated with diethyl ether and n-hexane to afford the required product 16 (0.75 g, 75.28%), mp 107-109° C.
Spectral and Elemental Analyses:
UVmax (MeOH): 242.4 nm (log ε 4.38) and 331.2 nm (log ε 4.30).
FTIRνmax (KBr): 2936.4, 1712.0, 1666.1, 1622.3, 1595.3, 1512.2, 1463.2, 1328.4, 1260.2, 1230.4, 1141.7, 1094.2, 1027.4, 917.7 and 810.6 cm−1.
1H NMR (CDCl3): δ 1.01 (s, 3H, 18-CH3), 1.25 (s, 3H, 19-CH3), 2.26 (m, 2H, —OCH2CH2CH2N<), 3.92 (s, 3H, —OCH3), 3.97 (t, 2H, —CH2N<), 4.24 (t, 2H, —OCH2—), 5.76 (s, 1H, 4-CH), 6.85 (d, 1H, Jo=8.30 Hz, 5-CH, aromatic), 6.94 (s, 1H, 5-CH, imidazole), 7.06 (m, 2H, 2-CH, aromatic and 4-CH, imidazole), 7.13 (dd, 1H, Jm=1.27 Hz, Jo=8.20 Hz, 6-CH, aromatic), 7.39 (s, 1H, vinylic-H, 16-arylidene) and 7.50 ppm (s, 1H, 2-CH, imidazole).
Calcd. For C33H40N2O4: C, 74.97; H, 7.63; N, 5.30. Found: C, 74.82; H, 7.66; N, 5.49.
16-[3-{3-(IMIDAZOL-1-YL)PROPOXY}BENZYLIDENE]-4-ANDROSTENE-3,1-DIONE (22) (DPJ-RG-1308) was prepared by Oppeneaur oxidation of 18 by a method as described for 16
EXAMPLE 3
a) 5-Androstene-3β,17β-diol
Sodium borohydride (0.06 g) was added in small portions to a solution of DHA (1 g, 3.47 mmol) in absolute alcohol (50 Ml) at room temperature. The resulting solution was cooled and then neutralized with dilute acetic acid. The solution was concentrated under reduced pressure and diluted with iced water. The curdy white precipitate obtained was crystallized from methanol to obtain the product 5-androstene-3β,17β-diol (0.9 g, 89.11%), mp 182-183° C. [Singh H., et al Indian J. Chem. 1969, 7, 1084-1087].
b) 5-Androstene-3β,17β-diol Diacetate
A mixture of 5-androstene-3β,17β-diol (1 g, 3.44 mmol), acetic anhydride (2 Ml) and dry pyridine (4 Ml) was heated in a steam bath for 2 h, and poured into iced water. The precipitate was filtered, washed with water, dried and crystallized in methanol to afford the product 5-androstene-3β,17β-diol diacetate (1 g, 81.3%), mp 163-165° C. [Singh H., et al Indian J. Chem. 1969, 7, 1084-1087].
c) 7-Oxo-5-androstene-3β,17β-diol Diacetate
A solution of 5-androstene-3β,17β-diol diacetate (5 g, 13.35 mmol) in a mixture of dry carbon tetrachloride (30 Ml), acetic anhydride (4.5 Ml) and glacial acetic acid (1.25 Ml) was treated while stirring with t-butyl chromate solution (35 Ml). The brown reaction mixture was brought to reflux temperature and further stirred for 12 h. The mixture was then cooled in an ice bath and treated in small portions with 10% oxalic acid solution (75 Ml). After adding a further quantity of solid oxalic acid (5.5 g), the reaction mixture was stirred for 2 h at room temperature. The organic phase was removed and the aqueous phase was extracted with carbon tetrachloride (4×50 Ml). The combined carbon tetrachloride extract was successively washed with 5% sodium carbonate solution (2×25 Ml), water and dried. Removal of solvent under reduced pressure gave a pale yellow solid, which was crystallized from methanol to afford the product 7-oxo-5-androstene-3β,17β-diol diacetate (3.2 g, 61.66%), mp 220-221° C. (lit 216-219° C.) [Singh. H., et al Indian J. Chem. 1969, 7, 1084-1087].
d) 7-Oximino-5-androstene-3β,17β-diol Diacetate
7-Oxo-5-androstene-3β,17β-diol diacetate (1 g, 2.57 mmol) and hydroxylamine hydrochloride (0.5 g) were taken in dry pyridine (8 Ml) and heated in a steam bath for 2 h. The reaction mixture was poured into water and precipitate obtained was filtered, washed with water and dried. The crude product on crystallization from methanol afforded pure oxime 7-oximino-5-androstene-3β,17β-diol diacetate (0.85 g, 86.54%), mp 236-238° C. (lit 226° C.) [Singh H., et al Indian J. Chem. 1969, 7, 1084-1087].
Spectral and Elemental Analyses:
1H NMR (CDCl3): δ 0.82 (s, 3H, 18-CH3), 1.14 (s, 3H, 19-CH3), 2.04 (s, 6H, 2× —OCOCH3), 4.67 (m, 2H, 3α-H and 17α-H), 6.46 (s, 1H, 6-CH) and 6.95 ppm (br, 1H, NOH, disappeared on adding D2O).
e) 7-[O-(3-Chloropropyl)oximino]-5-androstene-3β,17β-diol Diacetate (23) (DPJ-RG-1221)
A mixture of 7-oximino-5-androstene-3β,17β-diol diacetate (1 g, 2.48 mmol) and anhydrous potassium carbonate (2 g) was stirred and refluxed in ethyl methyl ketone (100 Ml) at 110° C. for 3 h. 1-Bromo-3-chloropropane (0.25 Ml, 2.48 mmol) was added to the reaction mixture and further refluxed for 20 h with continuous stirring. The completion of the reaction was monitored by TLC. The slurry was cooled, filtered and excess of solvent was removed under reduced pressure to obtain an oily residue. Iced water was added to the oily residue and it was allowed to stand overnight. The precipitate obtained was filtered, washed with water, dried and crystallized from pet-ether (60-80° C.) to afford 7-[O-(3-chloropropyl)oximino]-5-androstene-3β,17β-diol diacetate (23) (0.7 g, 58.82%), mp 155° C. (Decomp).
Spectral and Elemental Analyses:
UVmax (MeOH): 241.4 nm (log ε 4.16).
FTIRνmax (KBr): 2945.5, 2878.1, 1736.5, 1640.6, 1450.8, 1371.7, 1246.2, 1033.4, 953.2, 909.3 and 840.0 cm−1.
1H NMR (CDCl3): δ 0.81 (s, 3H, 18-CH3), 1.13 (s, 3H, 19-CH3), 2.03 (s, 6H, 2× —OCOCH3), 2.10 (m, 2H, —OCH2CH2CH2Cl), 3.60 (t, 2H, —CH2Cl), 4.14 (t, 2H, —OCH2—), 4.64 (m, 2H, 3α-H and 17α-H) and 6.43 ppm (d, 1H, 6-CH).
Calcd. For C26H38NO5Cl: C, 65.05; H, 7.98; N, 2.92. Found: C, 65.32; H, 8.18; N, 2.69.
f) 7-[O-{3-(Imidazol-1-yl)propyl}oximino]-5-androstene-3β,17μ-diol (25) (DPJ-RG-1223)
A mixture of 7-[O-(3-chloropropyl)oximino]-5-androstene-3β,17β-diol diacetate (23) (1 g, 2.08 mmol) and powdered imidazole (1.5 g) was fused at 110-120° C. for 5 h. The completion of reaction was monitored by TLC. The reaction was quenched with iced water to afford a the product 7-[O-{3-(imidazol-1-yl)propyl}oximino]-5-androstene-3β,17β-diol diacetate (24)
To the refluxing solution of 7-[O-{3-(imidazol-1-yl)propyl}oximino]-5-androstene-3β,17β-diol diacetate in methanol (50 Ml), potassium hydroxide (0.125 g) was added and the reaction mixture was further refluxed for 45 min. The reaction mixture was concentrated, acidified with glacial acetic acid and poured into cold water. The precipitate obtained was filtered, washed thoroughly with water, dried and crystallized from diethyl ether to furnish the product
25 (0.25 g, 23.45%), mp 205-206° C.
Spectral and Elemental Analyses:
UVmax (MeOH): 258.2 nm (log ε 4.17).
FTIRνmax (KBr): 3301.5, 3107.3, 2940.0, 2871.2, 1631.3, 1511.8, 1456.6, 1377.7, 1231.1, 1187.9, 1081.6, 1059.7, 1025.6, 952.0, 921.8, 889.0 and 823.3 cm−1.
1H NMR (CDCl3): δ 0.78 (s, 3H, 18-CH3), 1.13 (s, 3H, 19-CH3), 2.37 (m, 2H, —OCH2CH2CH2N<), 3.65 (m, 2H, 3α-H and 17α-H), 4.03 (m, 4H, —OCH2CH2CH2N<), 6.39 (d, 1H, 6-CH), 6.95 (brs, 5-CH, imidazole), 7.07 (brs, 4-CH, imidazole) and 7.49 ppm (brs, 2-CH, imidazble).
Calcd. For C25H37N3O3: C, 70.23; H, 8.72; N, 9.83. Found: C, 70.19; H, 8.94; N, 9.76.
EXAMPLE 4
a) 16α-Bromo-17-oxo-5-androsten-3β-ol
Cupric bromide (7 g) was added to a refluxing solution of 17-oxo-5-androsten-3β-ol (5 g, 17.33 mmol) in dry benzene (100 Ml) and dry methanol (100 Ml). The reaction mixture was further refluxed for 4 h and the reaction was monitored with TLC. The warm solution was filtered and concentrated to about two-third volume under reduced pressure. The remaining solvent was diluted with benzene (50 Ml) and washed twice with water. The benzene layer was collected, dried and the solvent removed under reduced pressure to obtain a residue, which was crystallized from methanol to afford the product 16α-bromo-17-oxo-5-androsten-3β-ol (4 g, 62.77%), mp 167-168° C. (lit 164-165° C.) [Glazier E R, J. Org. Chem., 1962, 2937-2938].
1H NMR (CDCl3): δ 0.93 (s, 3H, 18-CH3), 1.04 (s, 3H, 19-CH3), 3.55 (m, 1H, 3α-H), 4.55 (t, 1H, 16β-CH) and 5.38 ppm (m, 1H, 6-CH).
2-Bromo-1-Indanone (31) was prepared by bromination of 1-indanone with cupric bromide following the method as described by Glazier E R. J. Org. Chem., 1962, 2937-2938.
b) 16α/β-Bromo-4-androstene-3,17-dione (26)
16α-Bromo-17-oxo-5-androsten-3β-ol (1 g, 2.72 mmol) was dissolved in a mixture of cyclohexanone (10 Ml) and dry toluene (100 Ml). Traces of moisture were removed by azeotropic distillation of toluene. The distillation was continued at a slow rate while adding a solution of aluminium isopropoxide (1 g) in dry toluene (20 Ml) dropwise. The reaction mixture was refluxed for 1 h and allowed to stand at room temperature overnight. The slurry was filtered and the residue was washed with dry toluene. The combined filtrate and the washings were steam distilled until the complete removal of organic solvents was affected. The solid obtained was filtered, washed with water, dried and crystallized from a mixture of acetone and n-hexane to yield the product 16α/β-bromo-4-androstene-3,17-dione (26) (0.6 g, 60.33%), mp 128-130° C.
Spectral Analyses:
UVmax (MeOH): 238.8 nm (log ε 4.22).
IR (KBr): 2930.0, 1748.5, 1667.2, 1612.1, 1453.5, 1228.6 and 1017.9 cm−1 
1H NMR (CDCl3): δ 1.13 (s, 3H, 18-CH3), 1.22 (s, 3H, 19-CH3), 4.12 (t) and 4.56 (q) (3.5:1 area ratio, 1H, 16α-H and 16β-H) and 5.76 ppm (s, 1H, 4-CH).
c) 16β-(Imidazol-1-yl)-17-oxo-5-androsten-3β-ol (27) (DPJ-RG-1240)
A mixture of 16α-bromo-17-oxo-5-androsten-3β-ol (1 g, 2.72 mmol) and imidazole (1.5 g) was heated at 140-145° C. for 1 h. The reaction mixture was cooled to room temperature and precipitated with iced water. The solid obtained was filtered, washed with water, dried and crystallized from acetone to yield 27 (0.5 g, 51.81%), mp 249-251° C.
Spectral and Elemental Analyses:
FTIRνmax (KBr): 3303.8, 2943.0, 1744.8, 1499.5, 1456.1, 1374.7, 1318.3, 1243.9, 1065.8, 1041.4 and 914.2 cm−1.
1H NMR (CDCl3): δ 1.02 (s, 3H, 18-CH3), 1.07 (s, 3H, 19-CH3), 3.55 (m, 1H, 3α-H), 4.47 (m, 1H, 16α-H), 5.39 (d, 1H, 6-CH), 6.95 (s, 1H, 5-CH, imidazole), 7.12 (s, 1H, 4-CH, imidazole) and 7.70 ppm (s, 1H, 2-CH, imidazole).
Calcd for C22H30N2O2: C, 74.54; H, 8.53; N, 7.90. Found: C, 74.29; H, 9.04; N, 7.73.
2-(1H-1-imidazolyl)-1-indanone (32) was prepared by fusing imidazole with 31 as described for compound 27.
d) 16β-(Imidazol-1-yl)-5-androstene-3β,17β-diol (28) (DPJ-RG-1317)
To a stirred suspension of 16β-(imidazol-1-yl)-17-oxo-5-androsten-3β-ol (27), (1 g, 2.82 mmol) in methanol (100 Ml) at room temperature, sodium borohydride (1.5 g) was added in small fractions over a period of 2 h. The reaction mixture was further stirred for 4 h. Solvent was removed under reduced pressure and cold water was added. The precipitate obtained was filtered, washed with water, dried and crystallized from methanol to afford 28 (0.75 g, 74.57%), mp 285-287° C.
Spectral and Elemental Analyses:
FTIRνmax (KBr): 3259.0, 2930.9, 1501.4, 1459.5, 1365.1, 1235.8, 1152.7, 1083.2 and 955.1 cm−1.
1H NMR (CDCl3): δ 0.89 (s, 3H, 18-CH3), 1.03 (s, 3H, 19-CH3), 3.43 (m, 1H, 3α-H), 3.79 (m, 1H, 17α-H), 4.59 (q, 1H, 16α-H), 5.32 (d, 1H, 6-CH), 6.97 (s, 1H, 5-CH, imidazole), 7.04 (s, 1H, 4-CH, imidazole) and 7.59 ppm (m, 1H, 2-CH, imidazole).
Calcd for C22H32N2O2: C, 74.12; H, 9.05; N, 7.86. Found: C, 74.39; H, 9.62; N, 8.02.
e) 16β-(Imidazol-1-yl)-5-androstene-3β,17β-diol Diacetate (29) (DPJ-RG-1318)
A mixture of 16β-(imidazol-1-yl)-5-androstene-3β,17β-diol 28 (1 g, 2.80 mmol), acetic anhydride (2 Ml) and dry pyridine (2 Ml) was heated in a steam bath for 2 h. The reaction mixture was then poured into cold water and basified with liquid ammonia. The precipitate formed was collected by filtration, washed with water, dried and crystallized from acetone to afford 29 (0.8 g, 64.72%), mp 197-199° C.
Spectral and Elemental Analyses:
FTIRνmax (KBr): 2937.5, 1732.9, 1494.1, 1435.5, 1371.8, 1250.1, 1072.9, 1037.7 and 906.0 cm−1.
1H NMR (CDCl3): δ 1.02 (s, 3H, 18-CH3), 1.07 (s, 3H, 19-CH3), 1.74 (s, 3H, 17β-OCOCH3), 2.04 (s, 3H, 3β-OCOCH3), 4.61 (m, 1H, 3α-H), 4.79 (m, 2H, 16α-H and 17α-H), 5.39 (d, 1H, 6-CH), 6.89 (s, 1H, 5-CH, imidazole), 7.02 (s, 1H, 4-CH, imidazole) and 7.48 ppm (s, 1H, 2-CH, imidazole).
Calcd for C26H36N2O4: C, 70.88; H, 8.24; N, 6.36. Found: C, 70.29; H, 8.95; N, 6.49.
f) 16β-(Imidazol-1-yl)-4-androstene-3,17-dione (30) (DPJ-RG-1241)
A mixture of 16α/β-bromo-4-androstene-3,17-dione (26) (1 g, 2.74 mmol) and imidazole (1.5 g) was triturated and heated at 130-135° C. for 1 h. The reaction mixture was cooled at room temperature and cold water was added to it. The residue obtained was filtered, washed with water, dried and crystallized from a mixture of acetone and n-hexane to afford 30 (0.4 g, 41.45%), mp 179-181° C.
Spectral and Elemental Analyses:
UVmax (MeOH): 238.4 (log ε 4.31).
FTIRνmax (KBr): 2947.6, 1748.7, 1667.5, 1497.7, 1456.0, 1229.8, 1108.9, and 1045.0 cm−1.
1H NMR (CDCl3): δ 1.06 (s, 3H, 18-CH3), 1.24 (s, 3H, 19-CH3), 4.46 (m, 1H, 16α-H), 5.76 (s, 1H, 4-CH), 6.95 (s, 1H, 5-CH, imidazole), 7.09 (s, 1H, 4-CH, imidazole) and 7.66 ppm (s, 1H, 2-CH, imidazole).
Calcd for C22H28N2O2: C, 74.97; H, 8.01; N, 7.95. Found: C, 74.71; H, 8.58; N, 8.12.
Characterization
2-[2-(3-CHLOROPROPOXY)-BENZYLIDENE]-INDAN-1-ONE (3)
Yield=66.46%, mp: 130-134° C.
Anal:
IRνmax (KBr): 2970, 1680, 1600, 1440, 1230, 740 cm−1.
1H NMR (CDCl3): δ 2.31 (p, 2H, —OCH2CH2CH2Cl—), 3.81 (t, 2H, —CH2Cl), 3.98 (s, 2H, 3-CH2 indanone), 4.19 (t, 2H, —OCH2), 6.96 (d, 1H, Jo=8.49 Hz, aromatic), 7.03 (t, 1H, Jo=7.52 Hz, aromatic), 7.33 (dd, 1H, J Jo=6.83 Hz, aromatic), 7.40 (t, 1H, Jo=7.34 Hz, aromatic), 7.59 (m, 3H, aromatic □ndanones), 7.89 (d, 1H, Jo=7.61 Hz, 7-H □ndanones), 8.11 (s, 1H, vinylic —H).
Calcd. For C19H17O2Cl: C, 72.95; H, 5.47. Found: C, 70.31; H, 5.09.
2-[2-(3-IMIDAZOL-1-YL-PROPOXY)-BENZYLIDINE]-INDAN-1-ONE (4) (DPJ-1055)
Yield: 60.3%, mp: 76-78° C.
Anal:
IRνmax (KBr): 3360, 1680, 1600, 1140, 750 cm−1.
1H NMR (CDCl3): δ 2.31 (p, 2H, —OCH2CH2CH2—), 3.93 (t, 2H, —CH2N), 4.00 (s, 2H, 3-CH2 indanone), 4.26 (t, 2H, —OCH2), 6.87 (d, 1H, Jo=8.18 Hz, aromatic), 6.98 (d, 1H, imidazole), 7.39 (m, 8H, aromatic and imidazole), 7.92 (d, 1H, Jo=7.76 Hz, 7-H □ndanones), 8.19 (s, 1H, vinylic —H).
Calcd. For C19H17O2Cl: C, 72.95; H, 5.47. Found: C, 70.31; H, 5.09.
2-[3-(3-CHLOROPROPOXY)-4-METHOXYBENZYLIDENE]-INDAN-1-ONE (5)
Yield=42.86%, mp: 115-118° C.
Anal:
IRνmax (KBr): 2900, 1680, 1600, 1260, 1140, 730 cm−1.
1H NMR (CDCl3): δ 2.32 (p, 2H, —OCH2CH2CH2—), 3.81 (t, 2H, —CH2Cl), 3.88 (s, 3H, —OCH3), 3.96 (s, 2H, 3-CH2 indanone), 4.23 (t, 2H, —OCH2), 6.95 (d, 1H, Jo=8.3 Hz, aromatic), 7.21 (s, 1H, aromatic), 7.29 (dd, 1H, Jo=8.56 Hz, Jm=1.62 Hz, aromatic), 7.40 (t, 1H, Jo=7.10 Hz, aromatic □ndanones), 7.56 (m, 3H, aromatic □ndanones and vinylic-H), 7.88 (d, 1H, Jo=7.56 Hz, 7-H □ndanones).
Calcd. For C19H17O2Cl: C, 72.95; H, 5.47. Found: C, 70.31; H, 5.09.
2-[3-(3-IMIDAZOL-1YL PROPOXY)-4-METHOXYBENZYLIDENE)INDAN-1-ONE (6) (DPJ-RG-1090)
Yield: 55.49%, mp: 176-180° C.
Anal:
IRνmax (KBr): 2900, 1680, 1590, 1250, 1070, 740 cm−1.
1H NMR (CDCl3): δ 2.30 (p, 2H, —OCH2CH2CH2—), 3.93 (s, 3H, —OCH3), 3.96 (t, 2H, —CH2N), 4.01 (s, 2H, 3-CH2 indanone), 4.26 (t, 2H, —OCH2), 6.96 (d, 1H, Jo=8.45 Hz, aromatic and imidazolyl), 7.10 (d, 1H, aromatic), 7.33 (m, 2H, aromatic (□ndanones) and imidazolyl), 7.55 (m, 4H, aromatic (□ndanones), imidazolyl and vinylic-H) and 7.87 ppm (d, 1H, Jo=7.53 Hz, 7-CH □ndanones).
Calcd. For C23H22O3N2: C, 73.78; H, 5.92; N, 7.48. Found: C, 66.90; H, 5.93; N, 6.66.
2-[4-(3-CHLOROPROPOXY)-3-METHOXYBENZYLIDENE]-INDAN-1-ONE (7) (DPJ-1039)
Yield=27.5%, mp: 140-144° C.
Anal:
IRνmax (KBr): 2240, 1700, 1605, 1240, 1140 cm−1.
1H NMR (CDCl3): δ 2.33 (p, 2H, —OCH2CH2CH2—), 3.79 (t, 2H, —CH2Cl), 3.95 (s, 3H, —OCH3), 4.04 (s, 2H, 3-CH2 of □ndanones), 4.25 (t, 2H, —OCH2), 6.99 (d, 1H, Jo=8.41 Hz, aromatic), 7.20 (d, 1H, Jo=1.91 Hz, aromatic), 7.31 (dd, 1H, Jo=6.58 Hz, Jm=1.92 Hz, aromatic), 7.43 (t, 1H, Jo=6.96 Hz), 7.59 (m, 3H, aromatic □ndanones and vinylic-H), 7.91 (d, 1H, Jo=7.56 Hz, 7-H of □ndanones).
Calcd. For C20H19O3Cl: C, 70.07; H, 5.58. Found: C, 68.41; H, 4.93.
2-[4-(3-IMIDAZOL-1YL PROPOXY)-3-METHOXYBENZYLIDENE)INDAN-1-ONE (8) (RG-DPJ-195)
Yield: 34.28%; mp, 123-127° C.
Anal:
1H NMR (CDCl3): δ 2.33 (p, 2H, —OCH2CH2CH2—), 3.98 (s, 3H, —OCH3), 4.01 (t, 2H, —OCH2CH2CH2N), 4.29 (t, 2H, —OCH2—), 6.89 (d, 1H, aromatic), 6.99 (s, 1H, imidazolyl proton), 7.12 (s, 1H, aromatic), 7.21 (s, 1H, imidazolyl), 7.21 (s, 1H, aromatic), 7.44 (s, 1H, aromatic), 7.55 (t, 1H, aromatic) and 7.60 ppm (m, 3H, aromatic).
2-[3-(2-CHLOROETHOXY)-BENZYLIDINE]INDAN-1-ONE (9) (RG-DPJ-324)
Yield: 36.74%; mp 136-138° C.
Anal:
IR (KBr): 2955, 1680, 1590, 1420, 1225 and 730 cm−1.
1H-NMR (CDCl3): δ 3.88 (t, 2H, —CH2Cl), 4.06 (s, 2H, 3-CH2 indanone), 4.31 (t, 2H, —OCH2—), 6.96 (dd, 1H, Jo=8.12 Hz, Jm=2.52 Hz, aromatic), 7.22 (t, 1H, Jm=2.05 Hz, aromatic), 7.32 (d, 1H, aromatic), 7.40 (t, 2H, aromatic), 7.64 (d, 1H, aromatic), 7.71 (t, 1H, aromatic) and 7.91 ppm (d, 1H, 7-H □ndanones).
2-[3-(2-IMIDAZOL-1YL ETHOXY)-BENZYLIDINE]INDAN-1-ONE (10) (RG-DPJ-325)
Yield: 32.24%; mp 218-220° C.
Anal:
IR (KBr): 2940, 1690, 1610, 1240, 1085 and 740 cm−1.
1H-NMR (CDCl3): δ 4.04 (s, 2H, 3-CH2 indanone), 4.30 (t, 2H, —CH2N), 4.42 (t, 2H, —OCH2—), 6.92 (dd, 1H, aromatic Jo=8.10 Hz, Jm=2.36 Hz), 7.14 (m, 2H, aromatic), 7.36 (m, 3H, aromatic and imidazolyl), 7.51 (m, 3H, aromatic (□ndanones) and vinylic-H), 7.57 (s, 1H, imidazolyl) and 7.93 ppm (d, 1H, 7-CH □ndanones)
16-[3-(3-CHLOROPROPOXY)BENZYLIDENE]-17-OXO-5-ANDROSTEN-3β-OL (17) (DPJ-RG-1306)
Yield: 40.98%; mp, 145-147° C.
Anal:
FTIRνmax (KBr): 3419.2, 2932.9, 1715.6, 1628.7, 1580.1, 1445.1, 1375.0, 1257.2, 1215.4, 1163.3, 1054.2, 1008.0, 918.4 and 873.6 cm−1.
1H NMR (CDCl3): δ 0.90 (s, 3H, 18-CH3), 1.00 (s, 3H, 19-CH3), 2.19 (m, 2H, —OCH2CH2CH2Cl), 3.40 (m, 1H, 3α-H), 3.67 (t, 2H, —CH2Cl), 4.06 (t, 2H, —OCH2—), 5.27 (d, 1H, 6-CH), 6.79 (d, 1H, Jo=8.01 Hz, 4-CH, aromatic), 6.93 (s, 1H, 2-CH, aromatic), 7.02 (d, 1H, Jo=7.55 Hz, 6-CH, aromatic) and 7.18-7.24 ppm (m, 2H, 5-CH, aromatic and vinylic-H, 16-arylidene).
Calcd. For C29H37O3Cl: C, 74.26; H, 7.95. Found: C, 74.64; H, 8.12.
16-[3-{3-(IMIDAZOL-1-YL)PROPOXY}BENZYLIDENE]-17-OXO-5-ANDRO-STEN-3β-OL (18) (DPJ-RG-1307)
Yield: 45.87%; mp 211-212° C.
Anal:
FTIRνmax (KBr): 3253.0, 2932.8, 2837.0, 1713.9, 1629.0, 1576.6, 1448.6, 1385.6, 1269.0, 1160.8, 1067.0, 919.8, 872.7 and 779.6 cm−1.
1H NMR (CDCl3): δ 0.98 (s, 3H, 18-CH3), 1.07 (s, 3H, 19-CH3), 2.22 (m, 2H, —OCH2CH2CH2N<), 3.49 (m, 1H, 3α-H), 3.92 (t, 2H, —CH2N<), 4.21 (t, 2H, —OCH2—), 5.35 (s, 1H, 6-CH), 6.84 (dd, 1H, Jm=2.17 Hz, Jo=8.17 Hz, 4-CH, aromatic) 6.88 (s, 1H, 5-CH, imidazole), 7.02 (m, 2H, 2-CH, aromatic and 4-CH, imidazole), 7.13 (d, 1H, Jo=7.64 Hz, 6-CH, aromatic), 7.26-7.34 (m, 2H, 5-CH, aromatic and vinylic-H, 16-arylidene) and 7.44 ppm (s, 1H, 2-CH, imidazole).
Calcd. For C32H40N2O3: C, 76.77; H, 8.05; N, 5.59. Found: C, 76.85; H, 8.19; N, 5.65.
16-[3-{3-(IMIDAZOL-1-YL)PROPOXY}BENZYLIDENE]-17-OXO-5-ANDRO-STEN-3β-YL ACETATE (19) (DPJ-RG-1309)
Yield: 47.16%; mp, 148-150° C.
Anal:
FTIRνmax (KBr): 2940.6, 1728.2, 1631.8, 1575.8, 1508.0, 1443.8, 1374.8, 1254.0, 1163.7, 1030.5, 908.8 and 873.9 cm−1.
1H NMR (CDCl3): δ 0.98 (s, 3H, 18-CH3), 1.09 (s, 3H, 19-CH3), 2.03 (s, 3H, —OCOCH3), 2.23 (m, 2H, —OCH2CH2CH2N<), 3.91 (t, 2H, —CH2N<), 4.21 (t, 2H, —OCH2—), 4.58 (m, 1H, 3α-H), 5.41 (d, 1H, 6-CH), 6.83 (dd, 1H, Jm=2.15 Hz, Jo=8.11 Hz, 4-CH, aromatic) 6.87 (s, 1H, 5-CH, imidazole), 6.97 (s, 1H, 2-CH, aromatic), 7.02 (s, 1H, 4-CH, imidazole), 7.13 (d, 1H, Jo=7.73 Hz, 6-CH, aromatic), 7.30 (m, 2H, 5-CH, aromatic and vinylic-H, 16-arylidene) and 7.43 ppm (s, 1H, 2-CH, imidazole).
Calcd. For C33H42N2O4: C, 74.69; H, 7.98; N, 5.28. Found: C, 74.82; H, 7.49; N, 5.31.
16-[3-{3-(IMIDAZOL-1-YL)PROPOXY}BENZYLIDENE]-5-ANDROSTENE-3β,17β-DIOL (20) (DPJ-RG-1310)
Yield: 50.2%; mp, 133-135° C.
Anal:
FTIRνmax (KBr): 3373.7, 2936.1, 1601.2, 1438.1, 1366.7, 1252.4, 1161.9, 1050.8 and 955.5 cm−1.
1H NMR (CDCl3): δ 0.72 (s, 3H, 18-CH3), 1.04 (s, 3H, 19-CH3), 2.25 (m, 2H, —OCH2CH2CH2N<), 3.54 (m, 1H, 3α-H), 3.91 (t, 2H, —CH2N<), 4.06 (s, 1H, 17α-H), 4.21 (t, 2H, —OCH2—), 5.38 (d, 1H, 6-CH), 6.48 (d, 1H, vinylic-H, 16-arylidene), 6.73 (dd, 1H, Jm=2.29 Hz, Jo=8.03 Hz, 4-CH, aromatic), 6.87 (s, 4H, 2-CH, 6-CH, aromatic and 4-CH, 5-CH, imidazole), 7.26 (m, 1H, 5-CH, aromatic) and 7.48 ppm (s, 1H, 2-CH, imidazole).
Calcd. For C32H42N2O3: C, 76.46; H, 8.42; N, 5.57. Found: C, 76.42; H, 8.59; N, 5.41.
16-[3-{3-(IMIDAZOL-1-YL)PROPOXY}BENZYLIDENE]-5-ANDROSTENE-3β, 17β-DIOL DIACETATE (21) (DPJ-RG-1311)
Yield: 34.48%; mp, 97-99° C.
Anal:
FTIRνmax (KBr): 2939.5, 1732.6, 1600.5, 1440.4, 1372.0, 1241.2, 1162.1, 1034.7 and 907.6 cm−1.
1H NMR (CDCl3): δ 0.79 (s, 3H, 18-CH3), 1.05 (s, 3H, 19-CH3), 2.04 (s, 3H, 3β-OCOCH3), 2.21 (s, 3H, 17β-OCOCH3), 2.32 (m, 2H, —OCH2CH2CH2N<), 3.91 (t, 2H, —CH2N<), 4.20 (t, 2H, —OCH2—), 4.59 (m, 1H, 3α-H), 5.37 (s, 1H, 17α-H), 5.41 (d, 1H, 6-CH), 6.17 (s, 1H, vinylic-H, 16-arylidene), 6.73 (d, 1H, Jo=7.96 Hz, 4-CH, aromatic), 6.84 (s, 1H, 2-CH, aromatic), 6.93-7.06 (m, 3H, 6-CH, aromatic and 4-CH, 5-CH, imidazole), 7.26 (m, 1H, 5-CH, aromatic) and 7.49 ppm (s, 1H, 2-CH, imidazole).
Calcd. For C36H46N2O6: C, 73.69; H, 7.90; N, 4.77. Found: C, 73.51; H, 7.99; N, 4.94.
16-[3-{3-(IMIDAZOL-1-YL)PROPOXY}BENZYLIDENE]-4-ANDROSTENE-3,1-DIONE (22) (DPJ-RG-1308)
Yield: 50.25%; mp, 81-83° C.
Anal:
FTIRνmax (KBr): 2937.9, 1717.0, 1663.4, 1622.8, 1576.0, 1448.5, 1256.8, 1226.2, 1179.0, 1162.2, 1080.7, 1026.6, 915.8 and 864.3 cm−1.
1H NMR (CDCl3): δ 1.02 (s, 3H, 18-CH3), 1.25 (s, 3H, 19-CH3), 2.26 (m, 2H, —OCH2CH2CH2N<), 3.94 (t, 2H, —CH2N<), 4.23 (t, 2H, —OCH2—), 5.76 (d, 1H, 4-CH), 6.90 (dd, 1H, Jm=1.90 Hz, Jo=8.12 Hz, 4-CH, aromatic) 6.95 (s, 1H, 5-CH, imidazole), 7.02 (s, 1H, 2-CH, aromatic), 7.08 (s, 1H, 4-CH, imidazole), 7.16 (d, 1H, JO=7.68 Hz, 6-CH, aromatic), 7.33 (t, 1H, JO=7.91 Hz, 5-CH, aromatic), 7.40 (s, 1H, vinylic-H, 16-arylidene) and 7.57 ppm (s, 1H, 2-CH, imidazole).
Calcd. For C32H38N2O3: C, 77.08; H, 7.68; N, 5.62. Found: C, 77.22; H, 7.49; N, 5.88.
The synthetic routes for the preparation of various compounds have been depicted in schemes 1-5.




















Biological Activity

The enzyme was obtained from the microsomal fraction of freshly delivered human term placental tissue according to the procedure described in literature [Thompson, E. A.; Siiteri, P. K. J.; Biol. Chem., 1974, 249, 5364-5372]. The isolated microsomes were suspended in the minimum volume of phosphate buffer (0.05M, Ph 7.4) and stored at −30° C. as described. No loss of activity was observed within 4 months.
Inhibition of Aromatase. This assay was performed similar to described methods [Foster, A. B et al, J. Med. Chem. 1983, 26, 50-54; Graves, P. E et al Endocrinology 1979, 105, 52-57] monitoring enzyme activity by measuring the 3H2O formed from [1β,2β-3H] testosterone during aromatization. Each incubation tube contained 0.225 μCi of [1β,2β-3H] testosterone, 5 μM unlabeled testosterone, 2 Mm NADPH, 20 Mm glucose-6-phosphate, 1 EU glucose-6-phosphate dehydrogenase, and inhibitor (0-250 μM) in phosphate buffer (0.05M, Ph 7.4). The test compounds had been dissolved in EtOH and diluted with buffer. The final EtOH concentration of control and inhibitor incubation was 2%. Each tube was preincubated for 5 min at 30° C. in a shaking water bath. Microsomal protein (0.5 mg) was added to start the reaction. The total volume for each incubation was 0.5 Ml. The reaction was terminated by withdrawing 100-μL aliquots at 0, 7, 14 and 21 min and pipetting them into 200 μL of a cold 1 Mm HgCl2 solution. After addition of 200 μL of an aqueous dextran-coated charcoal (DCC) suspension (2%), the vials were shaken for 20 min and centrifuged at 1500 g for 5 min to separate the charcoal-adsorbed steroids. Aliquots of the supernatant were assayed for 3H2O by counting in a scintillation mixture in a Beckman liquid scintillation spectrometer (LS 8000).







TABLE 1







Aromatase inhibitory activity (IC50) of some valuable compounds










Compound


Inhibition


No.
Code
Structure
on CYP19a





 2
DPJ-RG- 1088




IC50: 0.7 Mm RP: 42.5


 


 4
DPJ-RG- 1055




IC50: 1.1 Mm RP: 26.6


 


 6
DPJ-RG- 1090




IC50: 1.3 Mm RP: 22.5


 


 8
RG-DPJ- 195




IC50: 0.55 Mm RP: 54.1


 


14
DPJ-RG- 1219




IC50: 2.4 Mm RP: 12.4


 


16
DPJ-RG- 1177




IC50: 4.4 μM RP: 6.8


 


20
DPJ-RG- 1310




IC50: 9.1 μM RP: 3.3


 


22
DPJ-RG- 1308




IC50: 4.4 μM RP: 6.8


 


24
DPJ-RG- 1315




IC50: 3.1 μM RP: 9.6


 


25
DPJ-RG- 1223




IC50: 12 μM RP: 2.5


 


27
DPJ-RG- 1240




IC50: 3.3 Mm RP: 9.0


 



DPJ-RG- 1241




IC50: 0.18 Mm RP: 165.3





a) Substrate: 1β 3H-androstenedione/androstenedione 500 nm


RP: Relative potency; relative to aminoglutethimide (=1).

















TABLE 2





Compound
Code
Melting point

Example


No.
number
(° C.)
% Yield
(Method)



















2
DPJ-RG-1088
48-50
46.59
1


4
DPJ-RG-1055
76-78
60.3
1


6
DPJ-RG-1090
176-180
55.49
1


8
RG-DPJ-195
123-127
34.28
1


10
RG-DPJ-325
218-220
32.24
1


12
DPJ-RG-1151
199-201
71.47
2


13
DPJ-RG-1196
109-111
50.04
2


14
DPJ-RG-1219
197-198
84.68
2


15
DPJ-RG-1227
163-168
65.64
2


16
DPJ-RG-1177
107-109
75.28
2


18
DPJ-RG-1307
211-212
45.87
2


19
DPJ-RG-1309
148-150
47.16
2


20
DPJ-RG-1310
133-135
50.20
2


21
DPJ-RG-1311
97-99
34.38
2


22
DPJ-RG-1308
81-83
50.25
2


24
DPJ-RG-1315
180-182
32.52
3


25
DPJ-RG-1223
205-206
23.45
3


27
DPJ-RG-1240
249-251
51.89
4


28
DPJ-RG-1317
285-287
74.57
4


29
DPJ-RG-1318
197-199
64.72
4


30
DPJ-RG-1241
179-181
41.45
4

## Claims

1. The imidazolyl 16-substituted-androst-4-ene-3,17-dione of the general formula A:

2. The imidazolyl 7-substituted-androst-5-ene steroidal derivatives of the general formula:

3. The imidazolyl 16-substituted steroidal derivative selected from the group consisting of:

4. The imidazolyl substituted steroidal derivative selected from the group consisting of:
16-[4-{3-(-Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-17-oxo-5-androst-en-3β-ol (12) (DPJ-RG-1151);
16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-17-oxo-5-androsten-3β-yl acetate (13) (DPJ-RG-1196);
16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-5-androstene-3β,17β-diol (14) (DPJ-RG-1219);
16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-5-androstene-3β,17β-diol diacetate (15) (DPJ-RG-1227);
16-[4-{3-(Imidazol-1-yl)propoxy}-3-methoxybenzylidene]-4-androstene-3,17-dione (16) (DPJ-RG-1177);
16-[3-{3-(Imidazol-1-yl)propoxy}benzylidene]-17-oxo-5-androstene-3β-ol (18) (DPJ-RG-1307);
16-[3-{3-(Imidazol-1-yl)propoxy}benzylidene]-17-oxo-5-androstene-3β-yl acetate (19) (DPJ-RG -1309);
16-[3-{3-(Imidazoi-1-yl)propoxy}benzylidene]-5-androstene-3β,17β-diol (20) (DPJ-RG-1310);
16-[3-{3-(Imidazol-1-yl)propoxy}benzylidene]-5-androstene-3β,17β-diol diacetate (21) (DPJ-RG-1311);
16-[3-{3-(Imidazol-1-yl)propoxy}benzylidene]-4-androstene-3,17-dione (22) (DPJ-RG-1308);
7-[O-{3-(Imidazol-1-yl)propyl}oximino]-5-androstene-3β,17β-diol diacetate (24) (DPJ-RG-1315);
7-[O-{3-(Imidazol-1-yl)propyl}oximino]-5-androstene-3β,17β-diol (25) (DPJ-RG-1223);
16β-(Imidazol-1-yl)-17-oxo-5-androsten-3β-ol (27) (DPJ-RG-1240);
16β-(Imidazol-1-yl)-5-and rostene-3β,17β-diol (28) (DPJ-RG-1317);
16β-(Imidazol-1-yl)-5-androstene-3β,17β-diol diacetate (29) (DPJ-RG-1318); and
16β-(Imidazol-1-yl)-4-androstene-3,17-dione (30) (DPJ-RG-1241).

5. The imidazolyl substituted steroidal derivative as claimed in any of claims 1-4 wherein the IC50 value for the aromatase inhibitory activity of the compound is ranging between 0.18-12 μM with relative potency of the compound ranging from 2.5 to 165.3 times as compared to standard drug aminoglutethimide.

6. A pharmaceutical composition comprising an effective amount of the imidazolyl substituted steroidal derivative as claimed in any of claims 1-4 or a salt thereof, and a pharmaceutically acceptable carrier, adjuvant, excipient, and/or diluent.