Quantification of the bombesin/gastrin releasing peptide antagonist RC-3095 by liquid chromatography-tandem mass spectrometry
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Updated: Aug 20, 2026
Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
Published on: August 17, 2015
Alberto S Pereira1, Luciane DiLeone, Fabiano H Souza
1Galeno Research Unit, Latino Coelho St., 1301, Parque Taquaral, 13087-010 Campinas, SP, Brazil.
Abstract:
Bombesin (BN) and its mammalian equivalent, gastrin-releasing peptide (GRP), stimulate cell proliferation and are involved in the pathogenesis of several types of human cancer. BN/GRP and their receptors were shown to be critical for the growth of various human malignancies, such as small-cell lung, prostate, ovary, stomach and breast cancers in the human tumor xenograft model. In the present study, a fast, sensitive, robust method was developed for the determination and quantification of a BN/GRP receptor antagonist RC-3095 (D-Tpi-Gln-Trp-Ala-Val-Gly-His-Leupsi(CH2NH)Leu-NH2), in human plasma by liquid chromatography coupled with tandem mass spectrometry. RC-3095 was extracted from 0.2 ml human plasma by protein precipitation using cold acetonitrile (0.4 ml). The method has a chromatographic run of 10 min using a C(8) analytical column (150 mm x 4.6 mm i.d.) and the linear calibration curve over the range was linear from 20 to 10000 ng ml(-1) (r(2)>0.994). The between-run precision, based on the relative standard deviation replicate quality controls, was 5.7% (60 ng ml(-1)), 7.1% (600 ng ml(-1)) and 6.8% (8000 ng ml(-1)). The between-run accuracy was +/-0.0, 2.1 and 3.1% for the above-mentioned concentrations, respectively. The developed procedure allows the quantitative determination of peptide RC-3095 for pharmacokinetics studies in human plasma.
Insights
A new method accurately quantifies the bombesin receptor antagonist RC-3095 in human plasma. This technique is crucial for understanding how RC-3095 works in pharmacokinetic studies for cancer treatment development.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Oncology
Background:
- Bombesin (BN) and gastrin-releasing peptide (GRP) promote cancer cell growth.
- BN/GRP receptors are key targets in various human cancers, including lung, prostate, ovary, stomach, and breast.
- RC-3095 is a novel antagonist targeting the BN/GRP receptor.
Purpose of the Study:
- To develop and validate a rapid, sensitive, and robust method for quantifying RC-3095 in human plasma.
- To establish a reliable analytical technique for supporting pharmacokinetic studies of RC-3095.
Main Methods:
- Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was employed.
- RC-3095 was extracted from human plasma via protein precipitation using cold acetonitrile.
- A C(8) analytical column was used with a 10-minute chromatographic run time.
Main Results:
- The method demonstrated excellent linearity over a wide concentration range (20-10000 ng/ml) with high correlation coefficients (r(2)>0.994).
- High precision was achieved, with between-run relative standard deviations ranging from 5.7% to 7.1%.
- Excellent accuracy was observed, with between-run deviations within +/-3.1% across tested concentrations.
Conclusions:
- A validated LC-MS/MS method enables precise and accurate determination of RC-3095 in human plasma.
- This method is suitable for quantitative analysis in pharmacokinetic studies, aiding cancer drug development.
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