design synthesis and in vitro screening of pyrazolines
play

Design, Synthesis and in vitro Screening of Pyrazolines based - PowerPoint PPT Presentation

Nov 30, 2023 •1.56k likes •1.8k views

Design, Synthesis and in vitro Screening of Pyrazolines based compounds as Phytohaemagglutinin ( PHA) mimetic Raj Kumar 1, *, Gaurav Joshi 1 , and Sandeep Singh 2 1 Laboratory for Drug Design and Synthesis, Centre for Pharmaceutical Sciences and

  1. Design, Synthesis and in vitro Screening of Pyrazolines based compounds as Phytohaemagglutinin ( PHA) mimetic Raj Kumar 1, *, Gaurav Joshi 1 , and Sandeep Singh 2 1 Laboratory for Drug Design and Synthesis, Centre for Pharmaceutical Sciences and Natural Products, Central University of Punjab, 151 001, Bathinda, India ; 2 Centre for Genetics Diseases and Molecular Medicine, Central University of Punjab, 151 001, Bathinda, India * Corresponding author: raj.khungher@gmail.com; rajcps@cup.ac.in 1

  2. Design, Synthesis and in vitro Screening of Pyrazolines based compounds as Phytohaemagglutinin ( PHA) mimetic 2

  3. Abstract: Phytohaemagglutinin (PHA, or phytohemagglutinin) is a lectin found commonly in plants, especially legumes. It has some physiological effects on cell metabolism; it induces mitosis and affects the cell membrane regarding transport and permeability to proteins. It agglutinates most mammalian red blood cell types and have the mitogenic effect. This is the reason that PHA is extensively used in the laboratory as well as clinical set up for karyotyping analysis. The downside of PHA use is its cost and storage (-20 o C) resulting into increased cost. We have synthesised acetylated pyrazolines as anticancer agents and during their evaluation for anticancer potential in normal control cells, we were surprised by their cell proliferation activity. We thought of relating our compounds to PHA (PHA mimetic) and performed the basis karyotyping experiment keeping PHA as standard and found our compounds to be PHA mimics. The compounds are thus being evaluated for their further PHA mimetic potential using B/T cell specific cell cycle analysis and karyotyping experiment keeping PHA as standard and found our compounds outstanding PHA in every aspect. The compounds are thus being evaluated for their further PHA mimetic potential. Keywords: Phytohaemagglutinin; Mitogenic; PHA mimetic 3

  4. Introduction  Phytohaemagglutinin (PHA, or phytohemagglutinin) is a lectin found in plants, especially legumes.  The lectin has a number of effects on cell metabolism; it induces mitosis, and affects the cell membrane in regard to transport and permeability to proteins. It agglutinates most mammalian red blood cell types.  Lymphocytes cultured with phytohaemagglutinin can be used for karyotype analysis 4

  5. Hypothesis  While evaluating a series of newly synthesized compounds on human peripheral blood monocytic cells, the synthesized compounds did not exhibit any cytotoxic activity but they showed increased proliferation indicated by the increased intensity of formazan reduction.  It was, thus, hypothesized that the synthesized compounds may be increasing the cell similar to PHA. 5

  6. Synthetic Strategy 6

  7. Chemical structures of the synthetics 7

  8. Objectives  Determining the absolute cell count in different samples and their comparison with PHA as control.  Determining the Protein Concentration of the tested samples against PHA.  Arresting the dividing cells at metaphase and observing them under microscope at 10x and 40x. 8

  9. Result and Discussions 9

  10. No. of cells were counted on Automated Cell Counter 70,000 60,000 50,000 40,000 30,000 20,000 10,000 0 PHA Control JP-9 JP-11 JP-12 JP-14 JP-16 JP-17 10

  11. Evaluation of cytotoxic effect on PBC in response to treatment with synthesized compounds at concentrations of 1 µL, 2 µL and 5 µL for a time duration of 72 hrs. Data is expressed as mean values ± S.D. of three independent experiments. 1µL 2µL 5µL 0.12 0.1 0.08 0.06 0.04 0.02 0 Control Vehicle Control PHA Control JP-7 JP-11 JP-20 JP-14 JP-21 JP-24 JP-30 JP-31 11

  12. Evaluation of cytotoxic effect on PBC in response to treatment with synthesized compounds at concentrations of 1 µL, 2 µL and 5 µL for a time duration of 96 hrs. Data is expressed as mean values ± S.D. of three independent experiments. 1µL 2µL 5µL 0.18 0.16 0.14 0.12 0.1 0.08 0.06 0.04 0.02 0 Control Vehicle Control PHA Control JP-7 JP-11 JP-20 JP-14 JP-21 JP-24 JP-30 JP-31 12

  13. Determination of Protein Concentration 45 40 35 30 25 20 15 10 5 0 JP-16 JP-17 JP-14 JP-12 PHA JP-11 JP-9 Control 13

  14. Protocol for Metaphase Arrest PHA C1 (JP-14) C2 (JP-21) Media (RPMI) 2mL 2mL 2mL Blood 150 µL 150 µL 150 µL PHA 30 µL C1 30 µL C2 60 µL 14

  15. Day1: Treatment Strategy Incubation for 68h at 37˚C 15

  16. Day2: Metaphase arrest and Harvesting of Cells 16

  17. Day 3 Slides preparation For PHA For compound C1 17

  18. Conclusions  Absolute cell count showed that JP-12 and JP-14 had increased cell count as compared to the control and PHA.  Protein concentration was also found to be considerably higher in the synthesized compounds in comparison to PHA.  Similar to PHA, JP-12 was able to bring about the metaphase arrest at the 68 th hour.  From this, it can be concluded that, JP-12 and JP-14, the synthesized compounds show cell proliferation as PHA and have the potential to replace PHA. 18

  19. Future Prospective  It would be more helpful to determine the cell type that is more susceptible to cell proliferation  Studies should be undertaken to delve into the mechanism of how the synthesized compounds bring about this effect. 19

  20. References 1. Hamelryck, T. W., Dao-Thi, M. H., Poortmans, F., Chrispeels, M. J., Wyns, L., Loris, R. (1996). The crystallographic structure of phytohemagglutinin-L. The Journal of Biological Chemistry . 271 (34): 20479 – 20485. 2. Mire-Sluis,A. R., Wickremasinghe, R. G., Hoffbrand,A. V., Timms, A. M. Francis, G. E., (1987). Human T lymphocytes stimulated by phytohaemagglutinin undergo a single round of cell division without a requirement for interleukin-2 or accessory cells. Immunology. 60 (1), 7 – 12. 3. Serke,S., Serke,M., Brudler, O. (1987). Lymphocyte activation by phytohaemagglutinin and pokeweed mitogen: Identification of proliferating cells by monoclonal antibodies. 99 (2), 167-172. 20

  21. Acknowledgements R.K. and SS thank DST and UGC, New Delhi, India for the financial assistance (F.No. SR/FT/CS-71/2011) and F.30-13/2013(BSR), respectively. Encouragement and support from Prof. P. Ramarao, Dean, and Prof. R.K Kohli, Vice Chancellor Central University of Punjab, is gratefully acknowledged. Special thanks are also due to Miss. Jimi Marin Alex, who thoroughly helped in accomplishing this task. 21

Download Presentation
Download Policy: The content available on the website is offered to you 'AS IS' for your personal information and use only. It cannot be commercialized, licensed, or distributed on other websites without prior consent from the author. To download a presentation, simply click this link. If you encounter any difficulties during the download process, it's possible that the publisher has removed the file from their server.

Recommend

Design, Synthesis and in vitro Biological Evaluation of Acrylamide Derivatives Against Chikungunya

Design, Synthesis and in vitro Biological Evaluation of Acrylamide Derivatives Against Chikungunya

Design, Synthesis and in vitro Biological Evaluation of Acrylamide Derivatives Against Chikungunya Virus Edeildo F. Silva-Jnior 1, *, Gabriel F.S. Passos 1 , Matheus G.M. Gomes 1 , Thiago M. Aquino 1 , Stephannie J.M. Souza 2 , Joo P.M.

390 views • 20 slides
In-vitro Screening for of Alcohalic and Hydroalcohalic Extracts of Ayurvedic Medicinal Plants for

In-vitro Screening for of Alcohalic and Hydroalcohalic Extracts of Ayurvedic Medicinal Plants for

In-vitro Screening for of Alcohalic and Hydroalcohalic Extracts of Ayurvedic Medicinal Plants for the management of hyperlipidemia Pooja Gaur, Karuna Shanker CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow-226015 Email:

444 views • 15 slides
Update on Research Using in vitro and Computer-based Tools for Screening Potential Estrogenic

Update on Research Using in vitro and Computer-based Tools for Screening Potential Estrogenic

Update on Research Using in vitro and Computer-based Tools for Screening Potential Estrogenic Activity Nov, 2008 - PPDC P. Schmieder EPA, ORD, National Health and Environmental Effects Research Laboratory Mid-Continent Ecology Division

679 views • 35 slides
Triazolylpyridazinones as a New Class of Antihypertensive Agents: Design, Synthesis and In vivo

Triazolylpyridazinones as a New Class of Antihypertensive Agents: Design, Synthesis and In vivo

Triazolylpyridazinones as a New Class of Antihypertensive Agents: Design, Synthesis and In vivo Screening Ravinesh Mishra 1, *, Anees Siddiqui 2 , Asif Husain 2 , Sweta Bhardwaj 2 and Mohd Rashid 2 1 School of Pharmacy and Emerging Sciences and

303 views • 16 slides
Preliminary Phytochemical Analysis and in vitro Anti-cancer Screening of Ethanolic extract of Roots

Preliminary Phytochemical Analysis and in vitro Anti-cancer Screening of Ethanolic extract of Roots

Preliminary Phytochemical Analysis and in vitro Anti-cancer Screening of Ethanolic extract of Roots of Bauhinia tomentosa L. against HUH-7 Human Liver cancer Cell lines Ramaiah Maddi 1* 1 Department of Pharmacognosy, Hindu College of Pharmacy,

411 views • 23 slides
Screening and Cancer Screening: more complex than we Cancer site Test(s) thought! Breast

Screening and Cancer Screening: more complex than we Cancer site Test(s) thought! Breast

5/26/16 A real world case: 39 year old woman undergoing in vitro fertilization (IVF) gets a mammogram as part of routine pre-IVF testing. The WISDOM of Screening She has no family history of cancer or personal history of breast disease.

416 views • 16 slides
Statistical methods for the objective design of screening procedures for design of screening

Statistical methods for the objective design of screening procedures for design of screening

Statistical methods for the objective design of screening procedures for design of screening procedures for macromolecular crystallization Daniel Hennessy, Bruce Buchanan, Devika Subramanian, Patricia A. Wilkosz and John M. Rosenberg

775 views • 13 slides
Pharmaceutical Cocrystals: An Emerging Approach to Physical Property Enhancement in the synthesis

Pharmaceutical Cocrystals: An Emerging Approach to Physical Property Enhancement in the synthesis

P harmaceutical during crystal form selection. Important design strategies for making cocrystals are described, along with some recent examples of using cocrystals to enhance specific physical properties. Cocrystal screening and synthesis are

191 views • 5 slides
Design for screening, emulation and calibration

Design for screening, emulation and calibration

Design for screening, emulation and calibration http://personalpages.manchester.ac.uk/staff/alexis.boukouvalas/ Joint work with Dan Cornford, Milan Stehl k, J.P. Gosling and H. Maruri-Aguilar beamer-aston-logo Design for screening,

401 views • 39 slides
1 On a more serious note... Theoretical foundations: The inhibition constant THERE ARE VERY GOOD

1 On a more serious note... Theoretical foundations: The inhibition constant THERE ARE VERY GOOD

Two basic types of experiments BATCH VS. SEQUENTI AL DESIGN OF ANY RESEARCH PROJECT Optimal Experiment Design design = choice of screening BATCH DESIGN OF EXPERIMENTS concentrations for Dose-Response Screening of Enzyme Inhibitors 1. decide

328 views • 7 slides
VHDL for Logic Synthesis Overview Design Flow for Hardware Design VHDL coding for

VHDL for Logic Synthesis Overview Design Flow for Hardware Design VHDL coding for

VHDL for Logic Synthesis Overview Design Flow for Hardware Design VHDL coding for synthesis General guidelines for hardware designers This lecture includes the content from: Nitin Yogi, Modelling for Synthesis with VHDL, Auburn

697 views • 52 slides
1 2 3 4 5 F I N A N C I A L P R O T E C T I O N S S E R V I C E SCREENING The Screening

1 2 3 4 5 F I N A N C I A L P R O T E C T I O N S S E R V I C E SCREENING The Screening

SCREENING GUIDE 1 2 3 4 5 F I N A N C I A L P R O T E C T I O N S S E R V I C E SCREENING The Screening Guide is based on a Scoping Study, which determined the Screening Tools and other relevant matters. See Chapter 1 for a summary of

480 views • 20 slides
Basic Synthesis Flow and Commands Technology Libraries Design Read/Write Design

Basic Synthesis Flow and Commands Technology Libraries Design Read/Write Design

Introduction to Digital VLSI Logic Synthesis Basic Synthesis Flow and Commands Technology Libraries Design Read/Write Design Objects Timing Paths Constraints Compile Wire Load Models Multiple

851 views • 67 slides
Logic Synthesis Overview Design flow Principles of logic synthesis Logic

Logic Synthesis Overview Design flow Principles of logic synthesis Logic

Logic Synthesis Overview Design flow Principles of logic synthesis Logic Synthesis with the common tools Conclusions 2 System Design Flow Electronic System Level (ESL) flow System C TLM, Verification, Profiling

1.12k views • 28 slides
Design and in-vitro testing of new antimicrobial peptides based on QSAR modelling Boris

Design and in-vitro testing of new antimicrobial peptides based on QSAR modelling Boris

Design and in-vitro testing of new antimicrobial peptides based on QSAR modelling Boris Vishnepolsky 1, *, Maya Grigolava 1 , George Zaalishvili 2 , Margarita Karapetian 2 and Malak Pirtskhalava 1, * 1 I.Beritashvili Center of Experimental

574 views • 30 slides
Hardware Design with VHDL Synthesis of VHDL Code ECE 443 Synthesis of VHDL Code This slide set

Hardware Design with VHDL Synthesis of VHDL Code ECE 443 Synthesis of VHDL Code This slide set

Hardware Design with VHDL Synthesis of VHDL Code ECE 443 Synthesis of VHDL Code This slide set covers Fundamental limitation of EDA software Realization of VHDL operator Realization of VHDL data type VHDL synthesis flow

511 views • 34 slides
Clinical Trials Integrating Immunotherapy and Radiation Therapy Karen Kelly, MD Professor of

Clinical Trials Integrating Immunotherapy and Radiation Therapy Karen Kelly, MD Professor of

Friday-Sunday, February 10-12, 2017, Miami, Florida Clinical Trials Integrating Immunotherapy and Radiation Therapy Karen Kelly, MD Professor of Medicine Associate Director for Clinical Research Jennifer Rene Harmon Tegley and Elizabeth Erica

494 views • 21 slides
Compositional and Distributional Models of Meaning for Natural Language Stephen Clark Natural

Compositional and Distributional Models of Meaning for Natural Language Stephen Clark Natural

Compositional and Distributional Models of Meaning for Natural Language Stephen Clark Natural Language and Information Processing Research Group University of Cambridge Computer Laboratory Oxford October 2010 C & C tools Intro Parsing

706 views • 59 slides
The Inequality We Want: How Much is Too Much? Alice Krozer - University of Cambridge Helsinki,

The Inequality We Want: How Much is Too Much? Alice Krozer - University of Cambridge Helsinki,

The Inequality We Want: How Much is Too Much? Alice Krozer - University of Cambridge Helsinki, September 5 th , 2014 UNU-WIDER conference Inequality measurement, trends, impacts and policy ROADMAP (A) To address inequality

192 views • 16 slides
Treatment Landscape in R/R DLBCL Novel Targets and Strategies Wyndham H. Wilson, M.D., Ph.D.

Treatment Landscape in R/R DLBCL Novel Targets and Strategies Wyndham H. Wilson, M.D., Ph.D.

Treatment Landscape in R/R DLBCL Novel Targets and Strategies Wyndham H. Wilson, M.D., Ph.D. Senior Investigator Gene-expression profiling of DLBCL subtypes Roschewski, M. et al. (2013) Nat. Rev. Clin. Oncol. Key signaling pathways in GCB DLBCL

1.15k views • 38 slides
ICLIO National Conference Care Coordination: General Perspectives on I-O Therapies Gary I.

ICLIO National Conference Care Coordination: General Perspectives on I-O Therapies Gary I.

ICLIO National Conference Care Coordination: General Perspectives on I-O Therapies Gary I. Cohen, MD Emeritus Chair, Berman Cancer Institute, GBMC Associate Professor, Oncology, SKCCC/JHU 9.30.16 Philadelphia, Pa. accc-iclio.org Cancer

236 views • 10 slides
Disclosures Update on COPD & Asthma No Pharma Disclosures NHLBI - Asthma Clinical

Disclosures Update on COPD & Asthma No Pharma Disclosures NHLBI - Asthma Clinical

Disclosures Update on COPD & Asthma No Pharma Disclosures NHLBI - Asthma Clinical Research Network Michael C. Peters, M.D. MAS Division of Pulmonary & Critical Care Medicine NHLBI Severe Asthma Research Program

399 views • 26 slides
Best Practices for a Successful Internship Program Diane Mellon, M.S.Ed. Coordinator of

Best Practices for a Successful Internship Program Diane Mellon, M.S.Ed. Coordinator of

Best Practices for a Successful Internship Program Diane Mellon, M.S.Ed. Coordinator of Experiential Learning Programs, USF Tampa Ben Heins, M.F.A. Coordinator of Internships and Service Learning, USF Sarasota-Manatee Content adapted with

507 views • 33 slides
Internships First Edition Comprehensive Guide to the entire process Swapneel Convert

Internships First Edition Comprehensive Guide to the entire process Swapneel Convert

Swapneels classic guide now updated and explained for 2017 Internships First Edition Comprehensive Guide to the entire process Swapneel Convert wanderlust Mehta Intern at IITB, into internships MSR, CERN The Part where

353 views • 31 slides

More recommend