Dr. Syed A. A. Rizvi
Department of Pharmaceutical Sciences, Hampton University School of Pharmacy (HUSOP), Flemmie P. Kittrell Hall 121 William R. Harvey Way Hampton, VA 23668, USA.

ISBN 978-93-5547-055-3 (Print)
ISBN 978-93-5547-056-0 (eBook)
DOI: 10.9734/bpi/caprd/v2

This book covers key areas of pharmaceutical research. The contributions by the authors include pharmacophore analysis, docking, carboxylesterases inhibitors, isatin derivatives, essential oil, antioxidant activity, cytotoxic effect, aflatoxin-producing fungal species, organic molecule, biomass valorization, biofuel development, colorectal cancer, buccal film, oral thin film, sublingual delivery system, gastro retentive, fast dissolving oral thin film, antimicrobial activity, anticancer activity, phytochemical analysis, minimum inhibitory concentration, Jurkat T-cell, acute lymphoblastic leukemia, phosphatidylserine, apoptosis, flow cytometry, cancer prevention and therapy, abnormal methylation enzymes, cancer stem cells, synthetic CDA formulations, differentiation inducers, differentiation helper inducers, antimicrobial activity, minimum inhibitory concentration, minimum bactericidal concentration, Alzheimer’s disease, dementia, solid lipid nanoparticles, in vitro cytotoxicity, pharmacokinetic study. This book contains various materials suitable for students, researchers and academicians in the field of pharmaceutical research.

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Isatin Derivatives as Human Intestinal Carboxylesterase Inhibitors: An Approach towards 3D-QSAR, Pharmacophore and Molecular Docking

Sanmati K. Jain, Piyush Ghode, Achal Mishra

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 1-21

Carboxylesterases (CEs) metabolize numeral drugs and their inhibitions extend the bioactivity or may decrease the harmfulness of compounds that are triggered by these enzymes. Isatin derivatives which were reported as carboxylesterase inhibitors selected in order to establish structure activity relationship quantitatively by using k-nearest neighbour molecular field analysis (kNN MFA). The dataset comprised of 49 compounds and sphere exclusion (SE) algorithm was applied for the division of the data set into training and test set. The models were generated using different values of dissimilarity levels, of which, the best model was obtained (dissimilarity value 3.75) showed cross validated correlation coefficient (q2) 0.8594 and predicted correlation (pred_r2) 0.8106 with test set of 9 compounds. kNN-MFA methodology with stepwise (SW) forward-backward, was used for building the QSAR models. The kNN-MFA contour plots showed relationship between structural features of substituted isatin derivatives and their activities which may be used to design newer potential CE inhibitors.

Pharmacophore studies reveals common two aromatic (AroC) and two hydrogen bond acceptors (HAc) features obtained from Molsign and Pharmagist approaches. The present work may be useful for further lead optimization and designing of potent carboxylesterase inhibitors. Molecular docking study was performed to identify potential interactions of the compounds with carboxylesterase active site. For this purpose the pdb id 2hrq (crystal structure of human carboxylesterase with soman) was chosen. Compound 62 exhibits a comparable docking score and bind into the active site of enzyme.

Study on Volatile Constituents, Cytotoxic Activity and Antioxidant Potential of Fixed Extracts of Copaifera luetzelburgii Harms

Sidney Goncalo de Lima, Juliana de Sousa Figueredo, Marcelo Costa dos Santos, Allan Kayk Sales Meneses, Marcio dos Santos Rocha, José Galberto Martins da Costa, Rozeverter Moreno Fernandes

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 22-36

Copaibas (Leguminosae - Caesalpinoideae) are native to the tropical region of Latin America and also West Africa. Their stem barks are used in folk medicine as a cicatrizing, anti-inflammatory, antiseptic, and antitumor agent. Thus, a phytochemical investigation of both the qualitative and quantitative chemical composition of the volatile and fixed constituents of the leaves, and bark of the Copaifera luetzelburgii stem is necessary and evaluated for their cytotoxicity, antioxidant potential and total phenol content. In this study, we aimed to evaluate the chemical composition of the volatile constituents, the cytotoxic activity, and the antioxidant potential against DPPH and ABTS radicals of C. luetzelburgii Harms, widely distributed in the Quilombola Community, municipality of Sao Miguel do Tapuio, State of Piauí, Brazil. The volatile constituents were extracted by the hydrodistillation method and analyzed by GC-MS. The ethanolic stem extract (EECC) was also analyzed by LC-ESI-TOF-MS. The phytochemical screening of the leaf and stem bark extracts (SBE) was performed, with the SBE showing more promising results.

While germacrene D (27.12%), \(\beta\)-caryophilylene (16.43%), and germacrene B (12.58%) were found to be the major constituents in the essential oil from leaves (EOL); \(\gamma\)-selinene (26.79%), 7-epi-\(\alpha\)-selinene (24.73%) and \(\beta\)-selinene (14.63%) were the major constituents in the oil of the stem bark. Toxicity evaluation was performed in Artemia salina with results considered inactive (LC50 >1000\(\mu\)g mL-1). The EECC showed the best of total phenols contents 31.41 ± 3.07, as, for antioxidant activity performed against the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) with CE50 value = 40.11 ± 2.31, while the ethanolic extract of copaiba leaves (EEFC) with CE50 value = 74.13 ± 6.05, all ready for the test 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) EECC showed better potential compared to EEFC. Thus, we hope that this work can contribute significantly to new research related to the species in phytochemistry and pharmacology.  

Determination of Antifungal and Anti-Aflatoxigenic Activity of the Essential Oil of Lippia lasiocalycina Cham (Verbenacea)

Wanessa Sales de Almeida, Sidney Gonçalo de Lima, Marcelo Costa dos Santos, Cândido Athayde Sobrinho, Rafael Gomes Abreu Bacelar, Maria Christina Sanches Muratori

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 37-51

Contamination of food by aflatoxin-producing fungal species generates huge economic losses and directly affects the health of consumers. The search for more natural preservation methods has raised interest in the potential of essential oils like antifungals in food. In this study, we aimed to evaluate the potential inhibition of Lippia lasiocalycina essential oil on fungal growth and aflatoxin B1 production by Aspergillus flavus strains. The minimum inhibitory concentration of the essential oil of L. lasiocalycina was determined against a strain of Aspergillus flavus. The potential for inhibition of fungal growth and aflatoxin B1 production was tested by direct contact and by the action of volatile compounds. Aflatoxin production was determined by HPLC. L. lasiocalycina essential oil inhibited 94% growth of Aspergillus flavus by direct contact and the volatile compounds potentiate the production of aflatoxin B1. The essential oil of Lippia lasiocalycina proved to be applicable in the production chain of aflatoxin B1 marketed as standard for use in scientific research.

Study on Emphasizing the Efficiency of a Cellulosolytic Preparation on the Hydrolyzed Wood Wastes

Ana- Despina Ionescu, Angela Casarica, Roxana - Madalina Stoica, Nicoleta Ene

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 52-60

According to the literature data, the cellulose, the most abundant organic molecule on Earth, is found mainly as a structural component of plants and algal cell walls, it is also produced by some animals such as tunicates, and it can be produced also by several bacteria, by a natural pathway or by biotechnology. Due to this situation, there are many studies focused on the use of this kind of renewable source and having as their issues the preparation of different valuable products, such as sugars, ethanol. Different chemicals, the whole yeasts biomass valorization with agricultural applications or for biofuels development. Our issue of this present study was to verify if we can avoid the microbial additional activity during the process, mostly while the conversion starts from some materials already pretreated, such as the paper and cellulose industry’s wastes, by using them as substrate in an optimized culture media formula, for the further yeasts strains development.

Molecular Docking and Pharmacokinetic Prediction Studies of Novel Coumarin Derivatives as Arylamine N- acetyltransferase 2 Inhibitors

Shiny George, P. K. Sujith, Meena Chandran, Kumaran Santhalingam

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 61-68

Arylamine N-acetyltransferases are xenobiotic-metabolizing enzymes responsible for detoxification of many drugs and carcinogens. It catalyzes the N- or O-acetylation of various arylamine and heterocyclic amine substrates and is able to bioactivate several known carcinogens. The levels of NATs in the body have important consequences with regard to an individual's susceptibility to certain drug-induced toxicities and cancers. Objective of present study is to analyse the affinity of computationally designed coumarin derivatives on NAT2 with AutoDock 4.0.1 software and to investigate the interactions between the target compounds and the amino acid residues of the enzyme. Molecular properties of designed compounds were studied by using Molinspiration. All compounds obeyed lipinski rule of five which suggest that these compound have excellent drug likeness properties and are preferable as an orally acting drug.  In present study, pharmacokinetic prediction on coumarin derivatives were done using pkCSM software. ADMET properties prediction results shown that all designed compounds possess well metabolic characteristics without obvious toxicities. Among the designed compounds 3, 6- dibutyl-7-hydroxy-4-oxo-2-chlrobenzyl -4H chromene -8- carbaldehyde (compound 5) shows more binding energy value (-9.08). These values suggested that the designed coumarin derivatives are excellent inhibitors of NAT2.

An Overview of Fast Dissolving Oral Thin Film

S. Muthukumar, R. Gayathri, A. Abarnadevika, C. Sanakr, Vinesha Ravi

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 69-77

Many pharmaceutical companies are switching their products from tablets to fast dissolving oral thin films (OTFs). Films have all the advantages of tablets (precise dosage, easy administration) and those of liquid dosage forms (easy swallowing, rapid bioavailability). Statistics have shown that four out of five patients prefer orally disintegrating dosage forms over conventional solid oral dosages forms. Pediatric, geriatric, bedridden, emetic patients and those with Central Nervous System disorders, have difficulty in swallowing or chewing solid dosage forms. Many of these patients are non-compliant in administering solid dosage forms due to fear of choking. OTFs when placed on the tip or the floor of the tongue are instantly wet by saliva. As a result, OTFs rapidly hydrate and then disintegrate and/or dissolve to release the medication for local and/or systemic absorption. This technology provides a good platform for patent non- infringing product development and for increasing the patent life-cycle of the existing products. The application of fast dissolving oral thin films is not only limited to buccal fast dissolving system, but also expands to other applications like gastroretentive, sublingual delivery systems.

Antioxidant and Anti-Osteosarcoma Effect of Edible Fungi Pleurotus ostreatus

J. Bindhu, E. Vihashinee, A. Rubiga

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 78-87

Osteosarcoma is a rare primary malignancy of bone and is extremely rare type, which accounts for about 0.2% of all cancer. It is treated with the advanced drugs which have effluent side effects when compared to the medicinal plants and phytoconstituents. The fundamental intention of this study is to study the antibacterial movement, phytochemical screening and anticancer action of ethanolic concentrate of Pleurotus ostreatus. The antimicrobial movement of Pleurotus ostreatus was recognized against two G -ve and two G +ve bacteria by agar well dissemination strategy. Based on the zone of inhibition formed the antimicrobial activity intensity were analyzed. The qualitative investigation of phytochemical screening was done which helps in analyzing the bioactive compounds present in the extract ethanolic and has demonstrated the nearness of flavonoids, terpenoids, alkaloids, saponins, starches, tannins, amino acids and steroids. The antioxidant test was done through DPPH and FRAP method. The invitro anticancer and cell toxicity of Pleurotus ostreatus against (osteosarcoma) human bone cancer cell lines compared to normal cell lines were premeditated. The anticancer test was experience against HL60 cancer cells of osteosarcoma. Subsequently, this investigation closes the ethanolic concentrate of Pleurotus ostreatus can be connected as a solid antimicrobial and anticancer properties.

Antimicrobial Studies: An Approach to Microbiological Assay

Sunil Kumar

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 88-99

This article includes brief introduction of bacteria and fungi along with commonly used bacteria and fungi which are used for antimicrobial activities. In this chapter, types of methods are also described in brief. This article includes how minimum inhibitory concentration (MIC) is measured in research work along with reported MIC of standard drugs for better result.

Purpose: The goal of this study is to develop discriminatory dissolution method to be used as a release parameter for testing and evaluating product performance by using quality by design trials intended for regulated markets. Dissolution testing is a requirement for all solid oral dosage forms and is used throughout the development life-cycle for product release and stability testing.

Methods: After testing solubility in the dissolution medium, surfactant concentration, rotation speed, dissolution medium pH, and apparatus type, the appropriate conditions were established.

Results: According to investigations, the optimal dissolution conditions were obtained utilising a USP apparatus II, 900 ml of 0.4 percent SLS medium in pH 4.5 acetate buffer, and a 75 rpm rotation speed.

Conclusion: This research shows how a discriminate dissolution method for Rivaroxaban, a BCS Class 2 medication with pH-independent solubility, was developed in a systematic manner. The dissolution method described here can be utilised as a quality control test for Rivaroxaban tablets, with a particular focus on batch to batch evaluations.

Role of PKC in CD47-Mediated Phosphatidylserine Expression Pathway in Jurkat T Cells: A Preliminary Study

Ikenna Kingsley Uchendu, Ogbonna Kosisochukwu Doris, Oguji Cyprian Emeka, Omeh Johnson Obiechina, Ochi John Chidubem, Udeh Abel Chukwuebuka, Obigeorge Chibueze Joseph, Chinecherem Maudlyne Nnam, Olatunji Omokungbe

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 107-119

Treatment failure in T-cell acute lymphoblastic leukaemia (T-ALL) occurs when leukemic blasts acquire resistance to chemotherapeutic agents. Current research efforts are focused on the search for targets for the development of more effective and less toxic anti-leukemic drugs. CD47 has been suggested to be involved in chemo-resistance and cell metastasis. Although several potential mechanisms were suggested to explain the therapeutic effect of CD47-targeting; the downstream effectors which lead to different effects by CD47 are still not well understood. In this preliminary study we assessed the role of Protein kinase C (PKC) in CD47-mediated phosphatidylserine expression pathway in jurkat T cells. Jurkat T cells were incubated with anti-CD47 mAb (BRIC 126), anti-CD44 mAb (BRIC 235) or control in the presence or absence of Bisindolylmaleimide I, hydrochloride (PKC inhibitor). Cells were stained with annexin-V FITC. Flow cytometry analysis was used for measurement of fluorescence intensity. Cell viability was detected using trypan blue exclusion test. PKC inhibition enhanced phosphatidylserine expression in CD47 receptor mediated leukaemia cells apoptotic pathway. This indicates that PKC may be involved in CD47- mediated PS exposure pathway in jurkat T cells. The observations from this study identify CD47 and PKC as novel functional proteins in jurkat T cells with promising therapeutic potential. This study would provide insight for targeted therapy against T- ALL disease.

Destabilization of Abnormal Methylation Enzymes as an Effective Therapeutic Strategy via Induction of Terminal Differentiation to Take Out Both Cancer Stem Cells and Cancer Cells

Ming C. Liau, Paul A. Fruehauf, Zhong-Hui Zheng, John P. Fruehauf

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 120-142

The objective of this study was to develop cancer drugs effective to take out both cancer stem cells (CSCs) and cancer cells, particularly CSCs since these cells stood in the way to deny the success of conventional therapies to put cancer away.

The association of methylation enzymes with telomerase constitutes a unique abnormality of cancer cells. This abnormality locks methylation enzymes in an exceptionally stable and active state so that hypomethylation of nucleic acids necessary for the cells to undergo Terminal Differentiation (TD) cannot take place. Human body produces metabolites that are able to eliminate telomerase from abnormal methylation enzymes of cancer cells to allow TD to proceed. Cell Differentiation Agent-2 (CDA-2) is a preparation of human metabolites from freshly collected urine, which has been approved for cancer therapy by the Chinese FDA. The effective components of CDA-2 are Differentiation Inducers (DIs) to target on the telomerase of abnormal methylation enzymes and Differentiation Helper Inducers (DHIs) which are the inhibitors of individual enzymes of ternary methylation enzymes. CDA-2 was very effective for the therapy of Myelodysplastic Syndrome (MDS), which is a disease attributable to Cancer Stem Cells (CSCs). We have previously carried out extensive studies on the DHIs of CDA-2.We are now focusing on the DIs of CDA-2 in order to formulate synthetic CDA for the prevention and therapy of cancer via targeting of CSCs.

DIs were purified from CDA-2 solution by procedures including differential solvent extraction, gel filtration, ion exchange chromatography, TLC, and HPLC. The mass of purified active preparation was determined by mass spectroscopy. DI activity was based on the Nitro Blue Tetrazolium (NBT) assay of HL-60 cells.

DIs of CDA-2 were found predominantly as acidic liposomal complexes extractable by dichloromethane. A good proportion of which became covalently linked to inactive carriers which were not soluble in dichloromethane, but soluble in alcohols. We have identified pregnenolone as a DHI of active liposomal complexes. After dissociation from pregnenolone, the active DIs of CDA-2 were not associated with UV absorption peaks of HPLC. We suspected that the active DIs might be acidic peptides derived from endogenous proteins, because we have previously found that acidic peptides of CDA-2 were active DIs. We, thus, randomly picked pentapeptides containing at least two acidic amino acid residues from the sequences of \(\alpha\)- and \(\beta\)-hemoglobin for synthesis to test their DI activities. Indeed, acidic pentapeptides of hemoglobin were active as DIs, although the activities were not impressive. Retinoic Acid (RA) and 12-O-TetradecanoylPhorbol-13-Acetate (TPA) are well known DIs with much better activities.

In this study, we found Pyrvinium Pamoate (PP) as the best DHI, and triinosinate + tetrainosinate (I3 + I4) as an acceptable DHI.

With effective DIs and DHIs on hand, our deliberated CDA formulations were as followings: for the therapy of MDS, the CDA-MDS formulation was RA(ED25)-5P-1(ED25)-I3 + I4(RI0.5)-PP(RI0.5)-sodium pregnenolone sulfate(RI0.5); for the therapy of CSCs, the CDA-CSC formulation was RA(ED25)- TPA(ED25)-PP(RI0.5)-resveratrol(RI0.5)-curcumin(RI0.5); for the therapy of brain tumor, the CDA-BT formulation was TPA(2xED25)-PP(2xRI0.5)-sodium phenylbutyrate(RI0.5)-pyrogallol(RI0.5); and for the therapy of melanoma and pancreatic cancer, the CDA-M&P formulation was RA(ED25)-TPA(2xED25)- 5P-1(ED25)-PP(2xRI0.5)-sodium tannate (RI0.5). The above CDA formulations all produced 100% NBT + on HL-60 cells. In conclusion, CDA formulations are the best to take out CSCs protected by drug resistant mechanism. A major biological mission of CSCs is to repair the wound. CDA formulations are primarily made up by DIs and DHIs which are wound healing metabolites, the partners involved in the biological mission of CSCs. Naturally, DIs and DHIs are tolerable to CSCs. CDA formulations can also induce cancer cells to undergo TD to terminate malignant growth.

Investigation on Phytochemical Constituents and Antimicrobial Properties of Crude Flower Extract of Tecoma stans (L.) Juss. ex Kunth

Sowjanya Pulipati, P. Srinivasa Babu

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 143-151

Since plants are used as therapeutic agents, the present study is designed to evaluate the phytochemical profile and antimicrobial activities of flower of Tecoma stans (L.) ethanolic extract (TSEE) against selective Gram-positive, Gram-negative bacteria in-vitro. Tecoma stans is a flowering perennial shrub belongs to the family Bignoniaceae. The entire plant possesses medicinal value and used for the treatment of various ailments.  The dried flowers were extracted by cold maceration process using ethanol. Presence of phytoconstituents such as alkaloids, glycosides, saponins, carbohydrates, tannins, phenolics compounds, steroids and flavonoids were observed. The antimicrobial activity was carried against bacteria like Staphylococcus aureus, Enterococcus faecalis, Bacillus subtilis, Bacillus megaterium, Streptococcus mutans, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Proteus vulgaris by means of agar well diffusion method.

The TSEE extract exhibited moderate to high level of broad-spectrum antimicrobial activities against these microorganisms. The diameters of zone of inhibition range from 20.33±0.57 to 09.33±1.52 mm; the activity was compared with standard tetracycline. MIC was performed by two-fold broth dilution method and the inhibition range was found to be 31.2 mg/ml to 125 mg/ml. The MBC values lies between 62.5 to 150 mg/ml. The results reported in the present work shows evidence that Tecoma stans flowers possess phytochemicals that exhibit broad spectrum antimicrobial activity against tested pathogenic bacteria.

Study on Utilization Pattern of Anti Dementia Drugs and Cost-Effective Analysis in India

S. Ray

Current Aspects in Pharmaceutical Research and Development Vol. 2, 18 September 2021, Page 152-160

Background: The population suffering from Alzheimer’s disease (AD) and other dementias is estimated to increase every year in India, the reason being steady growth in the older population and stable increment in life expectancy.

Aim: To evaluate the trend of clinical practices in India as well as cost analysis of anti dementia drugs in an Indian scenario.

Material and Methods: Utilization pattern of anti dementia drugs studies was studied using literature survey and data mining of Prospective and Retrospective studies. Cost analysis was performed to evaluate wide variation in the prices of same anti dementia drug with different formulation in India.

Results: Alzheimer’s disease is the most common subtype of dementia and Donepezil remains to be the most commonly prescribed drugs as indicated by Prospective and retrospective studies. Cost analysis of anti dementia drugs being manufactured by many companies across the different brands in India showed a very high variation in the minimum and maximum price. Cost ratio was highest for rivastigmine (1.5 mg capsule) and lowest for piracetam (400 mg tablet). It was observed that % cost variation of all the anti dementia drugs was above 100.

Conclusion: Thus, in India, a wide variation in the prices of same drug with different formulation has a severe economic impact. More effort is needed to understand and analyze in terms of prescription, utilization pattern of anti dementia drugs as it has huge socioeconomic impact on patients and caregivers of mainly developing countries like India.

Objective: Dasatinib (DST) is a Biopharmaceutics Classification System Class II drug having very low solubility and high permeability. Low aqueous solubility and poor dissolution of DST lead to poor bioavailability, Thus, limited aqueous solubility is the bottleneck for the therapeutic outcome of DST. Animal data suggest that the absolute bioavailability of DST is about 14–34% due to an extensive first-pass effect. To overcome hepatic first-pass metabolism and to enhance oral bioavailability, lipid-based drug delivery systems such as solid lipid nanoparticles (SLNs) can be used.

Methods: SLNs are submicron colloidal carriers having a size range of 50–1000 nm. These are prepared with physiological lipid and dispersed in water or aqueous surfactant solution. DST can be conveniently loaded into SLNs to improve the oral bioavailability by exploiting the intestinal lymphatic transport. An optimal system was evaluated for bioavailability study in rats compared with that of DST suspension (SUS).

Results: An in vitro cytotoxicity study was done by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay method through ATCC cell lines; the percent inhibition was more in SLN when compared with SUS. The pharmacokinetics of DST-SLNs after oral administration in male Wistar rats was studied. The bioavailability of DST was increased by 2.28 fold when compared with that of a DST SUS.

Conclusion: The results are indicative of SLNs as suitable lipid-based carrier system for improving the oral bioavailability of DST.

Natural products once served as the only source of medicines to mankind. Plants often synthesize unexpected novel structures which are biologically active, by various biosynthetic pathways. Phytochemical analysis of medicinal plants revealed the presence of various chemical constituents like alkaloids, flavonoids, Saponins, Glycosides, Triterpenoids, which possess a wide range  of Pharmacological activity. The Genus Crotalaria, family fabaceae includes 500 species distributed in tropical and subtropical regions and can be used as medical folklore remedies for treating different health disorders. Several flavones, chalcones and Chromeno chalcones have been isolated from these genus. A detailed re-investigation of Crotalaria ramisissima resulted in the isolation of a new dimethyl, dihydro chalcone Crotaramin, whose structure elucidation was determined by 2D spectral methods.