Metastatic Superscan in 18F PSMA PET/CT of a Patient with Prostate Carcinoma.

A biopsy-proven patient with prostate carcinoma aged 70 years was referred to the department of nuclear medicine for radionuclide-based therapy. His prostate-specific antigen levels were >1000 ng/mL, and prostatic magnetic resonance imaging showed an enlarged prostate with a heterogeneous signal and size 3.8x3.7x3.5 cm with few small heterogeneous nodular signals in the transition zone. He was scheduled for 18F prostate-specific membrane antigen (PSMA) positron emission tomography/computed tomography (PET/CT) scan before therapy. 18F PSMA PET/CT revealed PSMA-expressing prostate lesions (maximum standardized uptake value ~10.2) with extension into the urinary bladder along with bilateral supraclavicular, mediastinal, retrocrural, retroperitoneal, and pelvic lymph nodes and sclerotic lesions in the entire axial and appendicular skeleton.

Endobronchial Squamous Cell Carcinoma Presenting as Long Continuous Bronchial Thickening on 18Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography.

A 67-year-old man is presented with complaints of chest pain and productive cough for 1½ years. Chest X-ray was suggestive of right upper lobe Koch's lesion. Sputum was positive for mycobacterium tuberculosis. His symptoms got relieved partially by antitubercular treatment but the patient had an aggravation of symptoms for which he was evaluated. Computed tomography (CT) thorax revealed an endobronchial lesion in the right upper lobe bronchus. Bronchoscopy showed a mass in the right main bronchus and biopsy was suggestive of moderately differentiated squamous cell carcinoma (SCC). 18Fluoro-deoxy-glucose positron emission tomography/CT was performed for staging. There would have been chances of coexisting tuberculosis with SCC.

Synthesis and Antitumor Activity of Brominated-Ormeloxifene (Br-ORM) against Cervical Cancer.

Aberrant regulation of ß-catenin signaling is strongly linked with cancer proliferation, invasion, migration, and metastasis, thus, small molecules that can inhibit this pathway might have great clinical significance. Our molecular modeling studies suggest that ormeloxifene (ORM), a triphenylethylene molecule that docks with ß-catenin, and its brominated analogue (Br-ORM) bind more effectively with relatively less energy (-7.6 kcal/mol) to the active site of ß-catenin as compared to parent ORM. Herein, we report the synthesis and characterization of a Br-ORM by NMR and FTIR, as well as its anticancer activity in cervical cancer models. Br-ORM treatment effectively inhibited tumorigenic features (cell proliferation and colony-forming ability, etc.) and induced apoptotic death, as evident by pronounced PARP cleavage. Furthermore, Br-ORM treatment caused cell cycle arrest at the G1-S phase. Mechanistic investigation revealed that Br-ORM targets the key proteins involved in promoting epithelial-mesenchymal transition (EMT), as demonstrated by upregulation of E-cadherin and repression of N-cadherin, Vimentin, Snail, MMP-2, and MMP-9 expression. Br-ORM also represses the expression and nuclear subcellular localization of ß-catenin. Consequently, Br-ORM treatment effectively inhibited tumor growth in an orthotopic cervical cancer xenograft mouse model along with EMT associated changes as compared to vehicle control-treated mice. Altogether, experimental findings suggest that Br-ORM is a novel, promising ß-catenin inhibitor and therefore can be harnessed as a potent anticancer small molecule for cervical cancer treatment.

A Rare Case of Thyroid Cartilage Metastases Detected on 18F-Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography.

The thyroid cartilage metastatic involvement is an extremely rare entity. It can be asymptomatic at the earlier stage and can become symptomatic later on. Involvement of thyroid cartilage is frequent in melanoma and renal and rarely reported in an advanced stage of carcinoma prostate, breast, and lung. These cases were usually reported on positron emission tomography/computed tomography (PET/CT) as can often easily be missed on computed tomography scan alone. We present a case report of metastatic involvement of thyroid cartilage in squamous cell carcinoma of buccal mucosa detected on the whole-body 18F-fluorodeoxyglucose PET/CT.

Intracranial Dural Arteriovenous Fistulas with Cortical Venous Drainage: Radiosurgery as an Effective Alternative Treatment.

OBJECTIVE: To evaluate the clinical and radiological outcome of Gamma Knife radiosurgery (GKS) in treatment of intracranial dural arteriovenous fistula (DAVF) with cortical venous drainage (CVD) and compare it with the outcome of endovascular therapy. METHODS: Patients who underwent GKS or endovascular therapy for intracranial DAVF with CVD over 10 years (January 2007 to December 2016) at the All India Institute of Medical Sciences, New Delhi, were included. Demographics, clinical presentation, imaging details, and follow-up clinical status were reviewed retrospectively. Clinical follow-up was conducted once every 6 months. Radiological follow-up using digital subtraction angiography was performed at a mean 24 months after intervention. Patients with clinical follow-up of <1 year were excluded from the study. RESULTS: The study included 35 patients (26 in embolization group and 9 in GKS group) who had intracranial DAVF with CVD were included in the study. Clinical improvement was seen in 77.78% of the patients who received GKS and 57.7% of the patients who underwent embolization (P = 0.431). Complete obliteration of DAVF was seen in 55.56% of the patients in the GKS group and 57.7% of the patients in the embolization group (P = 1). GKS was at least as effective as embolization in terms of clinical and radiological outcome in treatment of intracranial DAVF with CVD. CONCLUSIONS: Contrary to popular perception, GKS should be considered as an effective first-line treatment alternative of intracranial DAVF with CVD.

Validation of ICH and ICH-GS Scores in an Indian Cohort: Impact of Medical and Surgical Management.

OBJECTIVE: Prognostic scores help in predicting mortality and functional outcome post intracerebral hemorrhage (ICH). We aimed to validate the ICH and ICH-GS scores in a cohort of Indian patients with ICH and observe the impact of any surgical intervention on prognostication. METHODS: This was an ambispective observational study of primary ICH cases enrolled between January 2014 and April 2018. Observed mortality on ICH and ICH GS scores for the entire cohort and individually for the medically and surgically managed patients was compared to the published mortality in the original derivation cohorts. RESULTS: 617 patients, (464 retrospective and 153 prospective) of ICH were included. In hospital mortality and 30-day mortality was 28.7% and 28.5% respectively. There was a significant association of increasing mortality with increasing ICH and ICH-GS scores. Area under receiver operating characteristic curve for 30-day mortality was 75.9% and 74.1% for ICH and ICH-GS scores respectively. However, mortality observed at individual scores was significantly less than previously reported. Among the surgically intervened patients (n = 265), both the expected mortality at baseline and discriminative ability of ICH and ICH-GS scores for 30-day mortality was significantly reduced following surgical intervention (ROC in surgically intervened groups: 59.9 (52.6-67.2) and 63(56-70) for ICH and ICH-GS scores respectively). CONCLUSIONS: Although ICH and ICH-GS scores are valid in Indian population, mortality at individual scores is lower than previously reported. Mortality prediction using ICH and ICH GS scores is significantly modified by surgical interventions. Thus, newer prognostic tools which incorporate surgical intervention need to be developed and validated in future.

Prevalence & factors associated with depression among schoolgoing adolescents in Chandigarh, north India.

BACKGROUND & OBJECTIVES: Depression among adolescents is a rising problem globally. There is a need to understand the factors associated with depression among adolescents. This study was conducted to ascertain the prevalence of depressive disorders and associated factors among schoolgoing adolescents in government and private schools in Chandigarh, India. METHODS: A cross-sectional study was conducted among 542 randomly selected schoolgoing adolescents (13-18 yr), from eight schools by multistage sampling technique. Depression was assessed using Patient Health Questionnaire-9 (PHQ-9) and associated factors by pretested semistructured interview schedule. Multivariate analysis was done to identify significant associated factors. RESULTS: Two-fifth (40%) of adolescents had depressive disorders, 7.6 per cent major depressive disorders and 32.5 per cent other depressive disorders. In terms of severity, 29.7 per cent had mild depression, 15.5 per cent had moderate depression, 3.7 per cent had moderately severe depression and 1.1 per cent had severe depression. Significant associated factors included being in a government school, studying in class Xth and XIIth, rural locality, physical abuse by family members, alcohol use and smoking by father, lack of supportive environment in school, spending less time in studies, lower level of participation in cultural activities and having a boy/girlfriend. Significant predictors on binary logistic regression analysis were being in class Xth [odds ratio (OR)=5.3] and lack of self-satisfaction with own academic performance (OR=5.1). INTERPRETATION & CONCLUSIONS: Our study showed that a significant proportion of schoolgoing adolescents suffered from depression. The presence of depression was associated with a large number of modifiable risk factors. There is a need to modify the home as well as school environment to reduce the risk of depression.

Ormeloxifene suppresses desmoplasia and enhances sensitivity of gemcitabine in pancreatic cancer.

The management of pancreatic ductal adenocarcinoma (PDAC) is extremely poor due to lack of an efficient therapy and development of chemoresistance to the current standard therapy, gemcitabine. Recent studies implicate the intimate reciprocal interactions between epithelia and underlying stroma due to paracrine Sonic hedgehog (SHH) signaling in producing desmoplasia and chemoresistance in PDAC. Herein, we report for the first time that a nonsteroidal drug, ormeloxifene, has potent anticancer properties and depletes tumor-associated stromal tissue by inhibiting the SHH signaling pathway in PDAC. We found that ormeloxifene inhibited cell proliferation and induced death in PDAC cells, which provoked us to investigate the combinatorial effects of ormeloxifene with gemcitabine at the molecular level. Ormeloxifene caused potent inhibition of the SHH signaling pathway via downregulation of SHH and its related important downstream targets such as Gli-1, SMO, PTCH1/2, NF-κB, p-AKT, and cyclin D1. Ormeloxifene potentiated the antitumorigenic effect of gemcitabine by 75% in PDAC xenograft mice. Furthermore, ormeloxifene depleted tumor-associated stroma in xenograft tumor tissues by inhibiting the SHH cellular signaling pathway and mouse/human collagen I expression. Xenograft tumors treated with ormeloxifene in combination with gemcitabine restored the tumor-suppressor miR-132 and inhibited stromal cell infiltration into the tumor tissues. In addition, invasiveness of tumor cells cocultivated with TGFß-stimulated human pancreatic stromal cells was effectively inhibited by ormeloxifene treatment alone or in combination with gemcitabine. We propose that ormeloxifene has high therapeutic index and in a combination therapy with gemcitabine, it possesses great promise as a treatment of choice for PDAC/pancreatic cancer.

Ormeloxifene efficiently inhibits ovarian cancer growth.

Ovarian cancer continues to be a leading cause of cancer related deaths for women. Anticancer agents effective against chemo-resistant cells are greatly needed for ovarian cancer treatment. Repurposing drugs currently in human use is an attractive strategy for developing novel cancer treatments with expedited translation into clinical trials. Therefore, we examined whether ormeloxifene (ORM), a non-steroidal Selective Estrogen Receptor Modulator (SERM) currently used for contraception, is therapeutically effective at inhibiting ovarian cancer growth. We report that ORM treatment inhibits cell growth and induces apoptosis in ovarian cancer cell lines, including cell lines resistant to cisplatin. Furthermore, ORM treatment decreases Akt phosphorylation, increases p53 phosphorylation, and modulates the expression and localization patterns of p27, cyclin E, cyclin D1, and CDK2. In a pre-clinical xenograft mouse ORM treatment significantly reduces tumorigenesis and metastasis. These results indicate that ORM effectively inhibits the growth of cisplatin resistant ovarian cancer cells. ORM is currently in human use and has an established record of patient safety. Our encouraging in vitro and pre-clinical in vivo findings indicate that ORM is a promising candidate for the treatment of ovarian cancer.

Assessment of raloxifene, estradiol-17ß, dl-ormeloxifene and levormeloxifene on thrombin activity.

Abstract Background: Cancer is one of the leading causes of morbidity and mortality globally. Cancer-associated thrombosis is well established in clinical settings, and thrombin has been found to induce angiogenesis at cancer sites. This establishes a link between cardiovascular diseases and cancer, where cancer and thrombin have been intricately associated. Various selective estrogen receptor modulators (SERMs) have been reported to exhibit anticancer activity. Therefore, we investigated estradiol-17ß and SERMs dl-ormeloxifene (centchroman), raloxifene and levormeloxifene (l-centchroman) for their anticancer effects and their effect on thrombin activity. Methods: Anticancer activity was assessed against PC-3 cell line by flow cytometry following treatment with estradiol-17ß and SERMs at 10 nM-1 mM concentrations. The cells were stained with propidium iodide and the percentage of cells in the sub-G0/G1 region was considered apoptotic. Thrombin inhibitory effect was evaluated by thrombin inhibition assay in vitro following incubation with 100 nM-3 mM concentrations of estradiol-17ß or various SERMs. Further, the effect of estradiol-17ß and SERMs on endogenous thrombin generation potential (ETP) was assessed by thrombin generation assay on rat plasma in vitro. Results: These compounds exhibited >90% cell death in PC-3 cell lines at 1 mM concentration except estradiol-17ß. Neither estradiol-17ß, dl-ormeloxifene and levormeloxifene showed any thrombin inhibitory or enhancing activity in thrombin inhibition assay, nor did they show any effect on ETP on rat plasma in vitro. However, raloxifene inhibited thrombin activity in a concentration-dependent manner. Raloxifene decreased ETP of the plasma at 3 and 1 mM,which is equivalent to that of 30-100 U/mL of heparin. Interestingly, raloxifene increased thrombin generation at lower concentrations and it inhibited thrombin generation at higher concentrations. Conclusions: These observations suggest that dl-ormeloxifene, estradiol-17ß and levormeloxifene do not possess thrombin inhibitory activity. Raloxifene possesses thrombin modulatory effect in addition to its anticancer activity, and this observation may help us in understanding the thromboembolic complications associated with raloxifene.

Relationship between bone turnover biomarkers, mandibular bone mineral density, and systemic skeletal bone mineral density in premenopausal and postmenopausal Indian women.

OBJECTIVE: Postmenopausal osteoporosis is one of the most common metabolic bone disorders. Osteoporosis is reported to cause bone loss in the alveolar processes of maxilla and mandible, which provide bony framework for tooth anchorage. However, the association between systemic osteoporosis and oral health remains controversial. Available evidence suggests that Indian women have lower peak bone mass than their Western/other Asian counterparts. The present study evaluated the relationship between mandibular bone mineral density (mBMD), systemic skeletal BMD, and bone metabolism in premenopausal and postmenopausal Indian women. METHODS: One hundred twenty-four premenopausal and 247 postmenopausal healthy women were included in the study. The BMD of the body of mandible, radius ultradistal, total hip, femur neck, and lateral spine were measured using dual-energy x-ray absorptiometry. Serum and urine biomarkers were determined using commercial kits. RESULTS: Univariate regression analysis followed by stepwise multivariate regression analysis to obtain the best fit model demonstrated the BMD of radius ultradistal, serum inorganic phosphorus, estradiol, and sex hormone-binding globulin as significant predictors of mBMD in premenopausal women. The BMD of femur neck, serum ionized calcium, bone-specific alkaline phosphatase, osteocalcin, and urine total pyridinoline were significantly associated with mBMD in postmenopausal women. The significant association between mBMD and number of teeth present was observed in the whole group of premenopausal and postmenopausal women. CONCLUSIONS: Varied predictors of mBMD were observed in premenopausal and postmenopausal women. The results suggest that the screening for these biomarkers and serum ionized calcium should be useful (1) to assess the status of mBMD particularly in women requiring surgical dental intervention that include bone manipulation and (2) for early detection and management of women with the risk of developing osteoporosis.

Epithelial-myoepithelial carcinoma of the lacrimal gland: a rare case.

Epithelial-myoepithelial carcinoma is a rare tumor of the salivary gland constituting only 1% of all tumors. It is a low-grade malignancy characterized by a classical biphasic morphology and immunophenotype. In the lacrimal gland, it is extremely rare with only 3 cases reported in the English medical literature. We describe the fourth case of epithelial-myoepithelial carcinoma, the first case in a female patient, and review the available literature.

Effect of ormeloxifene, a nonsteroidal once-a-week oral contraceptive, on systemic hemodynamics in adult female rats.

OBJECTIVE: To investigate the short-term effects of ormeloxifene on systemic hemodynamics, coagulation profile, and serum antioxidant activity in vivo in comparison with raloxifene. MATERIALS AND METHODS: Colony-bred adult female Sprague-Dawley rats were randomized into 19 groups of 10 each and received either ormeloxifene or raloxifene (0.25, 1.25, or 3 mg/kg/day) for 7, 15, or 30 days by the oral route. Animals of control group received vehicle (gum-acacia in distilled water) alone in a similar manner. Systemic hemodynamics and serum total antioxidant activity were assessed 24 h after the last treatment. RESULTS: There was no significant effect of ormeloxifene administered at these doses and schedules on hemodynamic parameters or antioxidant activity, except for increase in amplitude of R wave in rats treated with 3 mg/kg/day dose for 30 days. This effect with raloxifene was evident only 7 days after treatment at this dose. Overall response was, however, almost similar with both the agents. CONCLUSION: The findings demonstrate comparable pharmacological profile of ormeloxifene and raloxifene on short-term administration to rats. Based on changes observed in the ECG (R wave), long-term studies may lead to justifiable comparison of beneficial and harmful effects of ormeloxifene and raloxifene in relation to cardiovascular effects.

Osteogenic activity of constituents from Butea monosperma.

Phytochemical investigation from the stem bark of Butea monosperma, led to the isolation and identification of three new compounds named buteaspermin A (1), buteaspermin B (2) and buteaspermanol (3), along with 19 known compounds. The structure of compounds 1-22 were established on the basis of their spectroscopic data. The isolated compounds 2-17 were evaluated using neonatal (1-3 day old) rat calvaria derived primary osteoblast cultures. Five of these compounds 7, 10-13 showed promising osteogenic activity, attributed to increased osteoblast proliferation, differentiation and mineralization as evidenced by marked increase in expression of alkaline phosphatase, an early phase differentiation marker, and alizarin Red S staining of osteoblasts cultured for 48 h and von Kossa silver staining of nodules formed 15 days after culture with these compounds. Quantification of mineralization by optical density measurement of Alizarin Red S extracted from stained osteoblasts cultured for 7 days in presence of these compounds showed significant (P<0.05, vs corresponding vehicle control group) increase in mineralization. On the basis of biological results, structure-activity relationships are discussed.

Effect of ormeloxifene, a selective estrogen receptor modulator, on biomarkers of endometrial receptivity and pinopode development and its relation to fertility and infertility in Indian subjects.

OBJECTIVE: To compare expression of endometrial estrogen (ER) and progesterone (PR) receptors, beta(3)-integrin subunit, leukemia inhibitory factor (LIF), interleukin-6 (IL-6), and pinopodes in Indian women using ormeloxifene as a contraceptive with fertile and infertile subjects during the window of implantation. DESIGN: Controlled, prospective, clinical study. SETTING: Hospital-based reproductive health unit and research laboratories. PATIENT(S): Thirteen fertile women, 6 using ormeloxifene (30 mg/week), 29 with primary and 10 with secondary infertility. INTERVENTION(S): Transvaginal ultrasonography, midluteal endometrial biopsies, and blood samples. MAIN OUTCOME MEASURE(S): Histologic dating, endometrial thickness, immunohistochemical localization of ER, PR, beta(3)-integrin subunit, LIF, and IL-6 and scanning electron microscopy. RESULT(S): Ormeloxifene significantly reduced endometrial thickness, pinopode density, and caused histologic delay with increased epithelial ER and PR and unaltered epithelial beta(3)-integrin subunit expression. Appearance of fully developed pinopodes, down-regulation of epithelial PR, and increased epithelial beta(3)-integrin subunit expression was observed in in-phase endometrium from fertile and infertile women. LIF and IL-6 expression and serum estradiol and progesterone levels remained unaltered between groups. CONCLUSION(S): Ormeloxifene-induced effects might produce asynchrony between endometrial and embryo development resulting in implantation failure. Based on unaltered luteal phase serum progesterone concentration, ormeloxifene did not appear to prevent ovulation in any participant.

Postcoital interceptive activity of Wrightia tinctoria in Sprague-Dawley rats: a preliminary study.

BACKGROUND: This study was aimed to investigate the pregnancy-interceptive activity of the stem bark of Wrightia tinctoria R.Br. (Family Apocynaceae) administered during the preimplantation, peri-implantation and early postimplantation periods by oral route in adult female Sprague-Dawley rats. STUDY DESIGN: The ethanolic extract of the stem bark and its serial fractions were administered to female rats on Days 1-7 or 1-5 postcoitum (Day 1: day of sperm-positive vaginal smear) by the oral route. At autopsy on Day 10 postcoitum, the number and status of corpora lutea and implantations were recorded. For estrogen-agonistic activity, immature rats ovariectomized 7 days earlier received the test extract or the vehicle once daily for 3 days and, at autopsy on Day 4, uterine weight, status of vaginal opening and extent of vaginal cornification were recorded. RESULTS: The ethanolic extract of the stem bark of W. tinctoria R.Br. inhibited pregnancy in 100% of rats when administered orally at a 250-mg/kg dose on Days 1-7 or 1-5 postcoitum. On fractionation, the hexane-soluble, chloroform-soluble, water-soluble and water-insoluble fractions showed 100% anti-implantation effect, while n-butanol-soluble fraction intercepted pregnancy in 75% of animals when administered in the Days 1-5 postcoitum schedule. In immature rat bioassay, the active ethanolic extract and its fractions exhibited moderate to potent estrogen-agonistic activity, which might be responsible for their contraceptive action in this species. CONCLUSIONS: Findings demonstrate the antifertility activity of the ethanolic extract of the stem bark of W. tinctoria and its hexane-soluble, chloroform-soluble, water-soluble and water-insoluble fractions. Studies that pursue promising natural products (to identify contraceptive agents from natural sources lacking potent estrogenic activity) towards a fruitful conclusion for development/lead generation should continue.

Anti-Trichomonas activity of Sapindus saponins, a candidate for development as microbicidal contraceptive.

OBJECTIVES: Trichomoniasis is the most common non-viral sexually transmitted disease and is caused by the protozoan Trichomonas vaginalis. In view of increased resistance of the parasite to classical drugs of the metronidazole family, the need for new unrelated agents is increasing. This study evaluates anti-Trichomonas activity of Sapindus saponins, a component of a herbal local contraceptive Consap recently marketed in India. METHODS: The parasites were treated with saponins for MIC determination. Anti-Trichomonas activity of the saponins was evaluated using a cytoadherence assay, the substrate gel electrophoresis method and RT-PCR analysis. The effect of saponins on the mitochondrial potential of the host was determined by florescence-activated cell sorter. Actin cytoskeletal staining was used to determine the effect on parasite cytoskeleton. RESULTS: Using in vitro susceptibility assay, the MIC of Sapindus saponins for T. vaginalis (0.005%) was found to be 10-fold lower than its effective spermicidal concentration (0.05%). Saponins concentration dependently inhibited the ability of parasites to adhere to HeLa cells and decreased proteolytic activity of the parasite's cysteine proteinases. This was associated with decreased expression of adhesin AP65 and membrane-expressed cysteine proteinase TvCP2 genes. Saponins produced no adverse effect on host cells in mitochondrial reduction potential measurement assay. Saponins also reversed the inhibitory mechanisms exerted by Trichomonas for evading host immunity. Early response of saponins to disrupt actin cytoskeleton in comparison with their effect on the nucleus suggests a membrane-mediated mode of action rather than via induction of apoptosis. CONCLUSIONS: Findings demonstrate the potential of Sapindus saponins for development as a microbicidal contraceptive for human use. Further studies are required to evaluate its microbicidal activity against other sexually transmitted infections.

Synthesis and in vivo evaluation of 11-substituted estradiol derivatives as anti-implantation agents.

Synthesis of 11-substituted estradiol derivatives (12-17) has been carried out by the Grignard reaction with alkyl, allyl, and benzyl halides on 17beta-hydroxy-3-methoxy-11-oxo-estra-1,3,5(10),8(9)-tetraene (10). The novel compounds (10 and 12-17) were evaluated for their preliminary post-coital contraceptive (anti-implantation) activity in Sprague-Dawley rats. The tested compounds were administered orally and showed significant anti-implantation activity. Compound 13 is the most potent compound in the series which showed 100% contraceptive efficacy at 1.25 mg kg(-1).

Contraceptive and hormonal properties of the stem bark of Dysoxylum binectariferum in rat and docking analysis of rohitukine, the alkaloid isolated from active chloroform soluble fraction.

BACKGROUND: This study was aimed to investigate the pregnancy interceptive activity of the stem bark of Dysoxylum binectariferum Hook. f. administered during the pre- and peri-implantation periods and immediately after implantation by oral route in adult female Sprague-Dawley rats. STUDY DESIGN: Ethanolic extract and its fractions were administered to female rats on Days 1-10, Days 1-7, Days 1-5 or Day 1 postcoitum by oral route. At autopsy on Day 12, the number and status of corpora lutea and implantations were recorded. For estrogenic activity, ovariectomized immature rats received the test extract or the vehicle once daily for 3 days and at autopsy on Day 4, uterine weight and status of vaginal opening and extent of vaginal cornification were recorded. For antiestrogenic activity, the extract was administered along with ethinyl estradiol. Docking analysis of rohitukine, the alkaloid isolated from active chloroform soluble fraction, to estrogen receptor (ERalpha) was conducted using AutoDock 3.0.5 on a Linux workstation. RESULTS: The ethanolic extract intercepted pregnancy in rats at a daily dose of 500 mg/kg on Days 1-7 postcoitum. On fractionation, the activity was localized in the chloroform fraction, which inhibited pregnancy in all females at the 35-mg/kg dose on Days 1-7, at the 50-mg/kg dose on Days 1-5 or at the single 300-mg/kg dose on Day 1 postcoitum. Chromatography of this fraction yielded an alkaloid, rohitukine, which prevented pregnancy at the 10-mg/kg dose administered on Days 1-7 but was partially (45%) effective at this dose when administered during the entire preimplantation period and ineffective even at 10 times this dose when administered only on Day 1 postcoitum, except that there was a significant reduction in implantation number in pregnant females. While the active chloroform soluble fraction was devoid of any estrogen agonistic or antagonistic properties, a mild uterotropic effect without induction of premature opening of vagina or cornification of vaginal epithelium was observed in rohitukine at the 10-mg/kg dose. Rohitukine, with an almost similar molecular size (mol. wt. 305) as 17beta-estradiol, fits ideally into the hydrophobic pocket of ER. While it does not appear to simultaneously interact with GLU353, ARG394 and HIS524 as estradiol to elicit frank estrogenic response, different conformations of the ligand or its metabolite(s) might acquire geometry with phenolic groups at C-3', C-5 and C-7 positions disposed in a fashion to interact with active site(s) of ER, which might be responsible for its contraceptive and/or weak uterotropic effects. The absence of a basic side chain directed toward the antiestrogen binding site (ASP351) on the receptor appears to be responsible for the lack of any estrogen antagonistic activity. CONCLUSIONS: Findings demonstrate the antifertility activity of the ethanolic extract of D. binectariferum, its chloroform soluble fraction and rohitukine. Efforts are being made to enhance the anti-implantation activity of rohitukine by structural modifications.

Synthesis of 3-phenyl-4-phenylvinyl benzopyranones and the corresponding 2,2-dimethyl-benzopyrans with structural similarity to estradiol, as estrogen receptor ligands.

7-Methoxy-3-phenyl-4-phenylvinyl benzopyran-2-ones and the corresponding 2,2-dimethyl-benzopyrans, substituted with different alkylamino residues were synthesized. Except compound 13e, all compounds showed high level of estrogen agonistic activity (>81 %) whereas, compounds 13 b-e and 15a showed significant estrogen antagonistic activity (>20 %). X-Ray analysis of a 7-methoxy-3-phenyl-4-phenylvinyl benzopyran-2-one derivative 13d showed its structural resemblance to endogenous estrogen, 17beta-estradiol. Estrogenic and antiestrogenic activities of these derivatives demonstrate their estrogen receptor (ER) binding ability. The lack of hydroxyl groups at appropriate positions resulted in poor Relative Binding Affinity (RBA).