Specially for Natural Product

Contractual Services:

Bioactivity Guided Fractionation

Bioactivity guided fractionation & isolation of active principles:

Contractual Research, a wing of Natural Remedies is now offering, a service to isolate and identify bioactive constituents from herbal products. This new service has been the result of effective combination of Natural Remedies traditional strengths in phytochemistry with the battery of in-vitro bioassays (biochemical and cell based) standardized in-house.

Natural Remedies has taken up the initiative in isolating, purifying & characterizing over 200 different phytochemical constituents from important Indian medicinal plants to use as Primary Phytochemical Reference substance (www.phytocompounds.com). These reference substances have been characterized using chromatographic (TLC / HPTLC / HPLC), spectroscopic (UV, FTIR, NMR & Mass) and physico-chemical methods assuring their identity & purity.

Typical steps involved in Bioactivity guided fractionation:

  • Finalization of the bioassay based on the end use of the product (for example, DPP IV inhibition assay for healthy blood sugar claims)
  • Fractionation of the extract / product which will be guided by bioassay
  • Isolation of bioactive constituent(s) from the active fractions
  • Characterization of bioactive constituent(s) by spectroscopic (NMR, IR etc,.) methods
  • Chromatographic method (HPLC/HPTLC etc.,) for estimation of bioactive constituents in the product

What do you get from Bioactivity guided fractionation ?

  • Identification of bioactive constituent(s) of the herbs being used in your product
  • Reference standard of the bioactive constituent(s) of your product
  • Chromatographic method for estimation of bioactive constituent(s) in your product (can be used for meaningful quality control)

Natural remedies Bioassay Services

Biochemical assay:

Standardized / custom programs of preclinical discovery solutions to meet the needs of the Pharmaceutical, Natural Product and Biotechnology Industries. We have standardized and routinely perform more than 50 in-vitro assays relevant to selection and optimization of new drug candidates using specialized assay technologies based on different detection modes like Absorbance (colorimetric, ELISA), Luminescence and Fluorescence (Fluorescence Intensity, Fluorescence Polarization, Time Resolved Fluorescence etc) in 96 and 384 well formats. We provide a spectrum of these services in cellular assays that are designed to accelerate the pace and increase the productivity of discovery and development programs. We are committed in providing high quality, cost effective in vitro services with a rapid turnaround time. Apart from new product development, cell assays can be effectively be used for screening campaigns, bio-activity guided fractionation, compatibility studies, stability studies and for comparing various prototypes/formulations. In-vitro efficacy studies helps to screen the drug for its various therapeutic applications using high throughput model, thereby cost effective, repoducible results can be generated. In-vitro preclinical safety studies are standardized according to OECD guideline. These assays facilitates to explore the genotoxic potential of molecule.

Sl.No. code Bioassay Pharmacological Relevance
1 BCA01 α - Amylase inhibition Obesity, Type II Diabetes, Carbohydrate blocker
2 BCA02 α - Glucosidase inhibition Type II Diabetes, Carbohydrate blocker
3 BCA03 β - Glucuronidase inhibition Hepatoprotection, Axillary odour
4 BCA04 ABTS radical scavenging Anti-oxidant
5 BCA05 Acetylcholinesterase inhibition Cognition enhancement, Neuro- degenerative disorders
6 BCA06 Aryl Sulphatase inhibition Axillary odour
7 BCA07 Butyrylcholinesterase inhibition Neuro- degenerative disorders
8 BCA08 Collagenase  inhibition Antiageing / Antiwrinkle
9 BCA09 Nitric-oxide scavenging Anti-oxidant, Alzheimer’s disease
10 BCA10 Superoxide anion scavenging (PMS-NADH system) Anti-oxidant
11 BCA11 DPP IV inhibition Wrinkle, Ageing
12 BCA12 DPPH radical scavenging Inflammation, Anti-allergy
13 BCA13 Elastase inhibition Anti-oxidant
14 BCA14 Hyaluronidase inhibition Inflammation, Anti-allergy
15 BCA15 Hydroxyl radical scavenging Obesity
16 BCA16 Lipase inhibition Obesity
17 BCA17 15 - Lipoxygenase  inhibition Inflammation
18 BCA18 Trypsin inhibition Inflammation
19 BCA19 Xanthine oxidase inhibition Gout
20 BCA20 TC- PTPase inhibition Selectivity testing assay for anti-phosphatase activity
Cell assay:

Screening and assay development remains one of the most challenging and resource-intensive steps in the drug discovery process, with the end result depending on proper balance of assays, experimental design, and data management. Natural Remedies provides a broad spectrum of high quality service to support various stages of drug discovery and development. With state-of-the-art facilities and instrumentation, we offer services in assay development, compound profiling and in vitro safety. Natural Remedies uses a number of primary and immortalized cell lines to assay test compounds. We have 50 validated in vitro assays targeting various therapeutic areas using specialized assay technologies based on different detection models like absorbance (Colorimetric,ELISA), Luminescence and Fluorescence (Fluorescence Intensity, Fluorescence Polarization, Time Resolved Fluorescence) in 96 and 384 well formats. Using these in vitro cell based assays, compounds can be rapidly and cost effectively screened for activity prior to more expensive and time consuming in vivo analysis.

Safety assessment is an important criterion for compound development before expensive preclinical and clinical studies. In order to predict Genotoxicity or specific cellular impacts, Natural Remedies provides in vitro genetic testing (Ames, Chromosome Aberration, Micronucleus).

All work is carried out under well-defined standard operating procedures (SOPs) by highly qualified scientists. Most studies can be conducted under Good Laboratory Practice (GLP) compliance if required.

Sl.No. code Bioassay Pharmacological Relevance
1 CA01 Lymphocyte proliferation
a. Naïve and LPS treated lymphocytes
b. Naïve and PHA induced
c. Naïve and Con A treated lymphocytes
d. Naïve and PWA induced
e. Naïve and anti-CD3 induced		 Immune modulation
2 CA02 NO release Immune stimulation
3 CA03 NO scavenging Anti-oxidant, Inflammation, Pain
4 CA04 Macrophage phagocytosis Immune modulation
5 CA05 LTB4 release Inflammation, Pain
6 CA06 Hepatoprotection using t-BH Liver health
7 CA08 Histamine release Inflammation, Allergy
8 CA09 Chromosomal aberration assay Safety
9 CA10 Micronucleus assay Safety
10 CA11 Melanin release Skin Tanning / Lightning
11 CA12 PGE2 release Inflammation, Pain
12 CA13 Insulin release Diabetes
13 CA14 Collagen release Wrinkle, Ageing
14 CA15 TXB2 release Inflammation, Allergy, Pain
15 CA16 IL-1 beta inhibition Inflammation, Allergy
16 CA17 TNF alpha inhibition Inflammation, Allergy
17 CA18 AMES test Safety evaluation
18 CA19 IL-2 release (Con A induced) Immune modulation
19 CA20 IL-4 release (Con A induced) Immune modulation
20 CA21 IL-6 release (Con A induced) Immune modulation
21 CA22 IFN gamma release (Con A induced) Immune modulation
22 CA23 Neutral Red Uptake assay CC50 correlation to acute oral rodent LD50, safety evaluation
23 CA24 Cellular antioxidant assay Psoriasis
24 CA25 Hepatoprotection against Diazinon induced hepatotoxicity Liver health
25 CA36 IL-6 inhibition Inflammation
26 CA27 IL-6 release (PHA induced) Immune modulation
27 CA28 IL-6 release (PHA induced) Immune modulation
28 CA29 IL-2 release (PWA induced) Inflammation
29 CA30 IL-6 release (PWA induced) Immune modulation
30 CA31 IL-2 release (Anti CD3 induced) Immune modulation
31 CA32 IKK-β (NF-B kinase) inhibition IL-6 release (Anti CD3 induced)
32 CA33 IL-6 release (LPS induced) Immune modulation
33 CA34 IL-12 release (LPS induced) Immune modulation
34 CA35 Hepatoprotection assay against Aldrin (OC) Liver health
35 CA36 Adipogenesis assay Obesity
36 CA37 IL-3 release (Con A induced) Immune modulation
37 CA38 IL-5 release Con A induced) Immune modulation
38 CA39 IL-10 release (Con A induced) Immune modulation, Inflammation
39 CA40 IL-10 release (LPS induced) Immune modulation, Inflammation
40 CA41 Hepatoprotection against Aflatoxin(AFB1) induced hepatotoxicity in Human HepG2 cells Liver health
41 CA42 Hepatoprotection against Galactosamine induced hepatotoxicity in Human HepG2 cells Liver health
42 CA43 Adipolysis Obesity
43 CA44 MCP-1 (Monocytic chemotactic protein) inhibition Inflammation
44 CA45 MCF-7 anti-proliferative assay Cancer
List of Publications:

  • Modulation of lipopolysaccharide-induced pro-inflammatory mediators by an extract of Glycyrrhiza glabra and its phytoconstituents. Thiyagarajan P, Chandrasekaran CV, Deepak HB, Agarwal A. Inflammopharmacology. 2011. PMID: 21328091.
  • Optimization of cell-based assays to quantify the anti-inflammatory/allergic potential of test substances in 96-well format. Chandrasekaran CV, Edwin Jothie R, Kapoor P, Gupta A, Agarwal A. Inflammopharmacology. 2011 Jun;19(3):169-81. PMID: 21069571.
  • In vitro modulation of LPS/calcimycin induced inflammatory and allergic mediators by pure compounds of Andrographis paniculata (King of bitters) extract. Chandrasekaran CV, Thiyagarajan P, Deepak HB, Agarwal A. Int Immunopharmacol. 2011 Jan;11(1):79-84. PMID: 21034865.
  • Dual inhibitory effect of Glycyrrhiza glabra (GutGard™) on COX and LOX products. Chandrasekaran CV, Deepak HB, Thiyagarajan P, Kathiresan S, Sangli GK, Deepak M, Agarwal A. Phytomedicine. 2011 Feb 15;18(4):278-84. PMID: 20864324.
  • Effect of an extract of Andrographis paniculata leaves on inflammatory and allergic mediators in vitro. Chandrasekaran CV, Gupta A, Agarwal A. J Ethnopharmacol. 2010 May 27;129(2):203-7. PMID: 20307638.
  • In vitro efficacy and safety of poly-herbal formulations. Chandrasekaran CV, Sundarajan K, David K, Agarwal A. Toxicol In Vitro. 2010 Apr;24(3):885-97. PMID: 19958825.
  • Tinospora cordifolia, a safety evaluation. Chandrasekaran CV, Mathuram LN, Daivasigamani P, Bhatnagar U. Toxicol In Vitro. 2009 Oct;23(7):1220-6. PMID: 19651204.
  • Evaluation of the genotoxic potential and acute oral toxicity of standardized extract of Andrographis paniculata (KalmCold). Chandrasekaran CV, Thiyagarajan P, Sundarajan K, Goudar KS, Deepak M, Murali B, Allan JJ, Agarwal A. Food Chem Toxicol. 2009 Aug;47(8):1892-902. PMID: 19447157.

ADME assay:

As researchers try to predict the absorption, distribution, metabolism and excretion (ADME) characteristics of a compound, the advent of various in-vitro high throughput assays has come as a boon. The liver is the central organ in ADME. It is responsible for most of the metabolic processes involving break down of drugs and their excretion from the body. The cytochrome P450 superfamily of enzymes sits at the crossroads of liver metabolism, encompassing dozens of gene products that catalyze the breakdown of drugs and toxins. We at Natural Remedies have standardized several Cytochrome P450 assays that are useful for studying interaction as well as metabolism. In addition we have standardized various in-vitro permeability models like PAMPA and Caco-2 monolayer permeability assay that are useful for absorption studies.

Sl.No. code Bioassay Pharmacological Relevance
1 AM01 Cytochrome P450 2A5 (m) inhibition Interaction
2 AM02 Cytochrome P450 2A6 (h) inhibition Interaction
3 AM03 Cytochrome P450 3A4 (H) inhibition Interaction
4 AM04 Cytochrome P450 2D6 (h) inhibition Interaction
5 AM05 Glutathione S-transferase (GST) (r) inhibition Interaction
6 AM06 UDP Glucuronosyltransferase (UGT) (h) inhibition Interaction

Disclaimer

The in-vitro bioassays given in this list / catalogue are being offered under company’s contract research program. In-house standardized assay protocols will be used to perform these assays. The company is committed to ensure that utmost care is taken to adhere to specified protocols towards providing accurate and factual results to the users of this service. However, further use of the results and analytical reports provided by Natural Remedies for any commercial purposes is the sole responsibility of the user and Natural Remedies does not make any representation that the use of the bioassay reports will not infringe any intellectual property rights of third parties. In no event shall Natural Remedies be liable to any party for any direct, indirect, special or other consequential damages for any commercial use of the bioassay reports provided by Natural Remedies.

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