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Assay for the Detection of the Phenylpiperazine Family

a technology of phenylpiperazine and detection method, which is applied in the field of piperazine derivatives, can solve the problems of expensive equipment and highly trained staff, and the detection level of this kit is insufficient for testing purposes

Inactive Publication Date: 2013-08-29
RANDOX LAB LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes new substances that can be used to make antibodies that can detect various compounds related to phenylpiperazine. These antibodies are better and faster than existing methods, making them useful for detecting these compounds in samples. This invention is cost-effective and easy to use.

Problems solved by technology

Coupled with their ease of availability and varying legal status across the globe this can create the misconception that these drugs are safe and without the risks commonly associated with traditional street drugs.
A disadvantage of mass spectrometry-based methods of detection is that they require expensive equipment and highly trained staff.
The manufacturers include no information for cross-reactivity with mCPP or TFMPP in kit inserts and detection levels of this kit are insufficient for testing purposes.

Method used

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  • Assay for the Detection of the Phenylpiperazine Family
  • Assay for the Detection of the Phenylpiperazine Family
  • Assay for the Detection of the Phenylpiperazine Family

Examples

Experimental program
Comparison scheme
Effect test

example 1

Conjugation of 3-(carboxyphenyl)piperazine (Hapten-A) to BSA (to form Immunogen-I)

[0065]To a solution of 3-(carboxyphenyl)piperazine (Hapten-A) (30.9 mg, 0.15 mM) in DMF (1.0 ml) was added N,N-dicyclohexylcarbodiimide (DCC) (33.08 mg, 0.16 mM) and N-hydroxysuccinimide (18.7 mg, 0.16 mM) and the mixture was stirred at room temperature overnight. The dicyclohexylurea formed was removed by filtration and the solution was added dropwise to a solution of BSA (150 mg, 2.3 mmol) in 50 mM sodium bicarbonate solution (pH 8.5) (10 ml). The mixture was then stirred overnight at 4° C. The solution was then dialysed against 50 mM phosphate buffer pH 7.2 (3 changes) for 24 hours at 4° C., and freeze-dried to give Immunogen-I.

[0066]MALDI results showed 14.07 molecules of Hapten-A had been conjugated to one molecule of BSA.

example 2

Conjugation of 3-(carboxyphenyl)piperazine (Hapten-A) to BTG (to form Immunogen-II)

[0067]To a solution of 3-(carboxyphenyl)piperazine (Hapten-A) (41.8 mg, 0.203 mmol) in DMF (1.0 ml) was added N,N-dicyclohexylcarbodiimide (DCC) (46.01 mg, 0.223 mmol) and N-hydroxysuccinimide (25.66 mg, 0.223 mmol) and the mixture was stirred at room temperature overnight. The dicyclohexylurea formed was removed by filtration and the solution was added dropwise to a solution of BTG (150 mg, 2.25 μmol) in 50 mM sodium bicarbonate solution (pH 8.5) (10 ml). The mixture was then stirred overnight at 4° C. The solution was then dialysed against 50 mM phosphate buffer pH 7.2 (3 changes) for 24 hours at 4° C., and freeze-dried to give Immunogen-II.

example 3

Conjugation of 3-(carboxyphenyl)piperazine (Hapten-A) to HRP

[0068]EDC hydrochloride (10 mg) was dissolved in water (0.5 ml) and immediately added to a solution of 3-(carboxyphenyl)piperazine (Hapten-A) (2 mg) in DMF (0.2 ml). After mixing, this solution was added dropwise to a solution of HRP (20 mg) in water (1 ml). Sulfo-NHS (5 mg) was added and the reaction mixture was incubated in the dark at room temperature overnight. Excess hapten was removed with double PD-10 columns (Pharmacia) in series, pre-equilibrated with PBS at pH 7.2. The hapten-A-HRP conjugate was then dialysed overnight against 10 L of PBS at pH 7.2 at 4° C.

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Abstract

The current invention describes novel immunogens which are used in the production of novel antibodies with unique binding properties in that they cross-react with a variety of phenylpiperazine derivatives. These antibodies enable methods and kits to detect and / or determine phenylpiperazine derivatives (for example mCPP, TFMPP and MeOPP) in an in vitro sample which are advantageous over currently available analytical methods in terms of cost, ease of use, speed and sensitivity.

Description

BACKGROUND[0001]In recent years piperazine derivatives have emerged as a new class of designer drugs, gaining popularity especially among young people in the dance music scene where they are commonly known as ‘party pills’. They are often marketed as ‘natural’ or ‘herbal’ products but are in fact entirely synthetic chemicals. Coupled with their ease of availability and varying legal status across the globe this can create the misconception that these drugs are safe and without the risks commonly associated with traditional street drugs. Piperazine derivatives are usually found in illicit dosage forms as either tablets or capsules, but loose powders and more rarely solutions also occur. The tablets often carry logos similar to those seen on ecstasy tablets and they are often misleadingly sold as ecstasy or as apparent ‘safer’ and ‘legal’ alternatives. Their name derives from the piperazine heterocycle which is a common feature and they can be divided into two sub-classes; the benzylp...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07D241/04
CPCC07D295/096C07D241/04C07D295/155C07K14/47C07K14/765C07K16/44C07K2317/76G01N33/5308
Inventor BENCHIKH, ELOUARDMCCONNELL, IVANLOWRY, PHILIPFITZGERALD, PETER
Owner RANDOX LAB LTD
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