Selcia Radiolabelling

Selcia Radiolabelling

14C Custom Radiosynthesis 14C GMP API for Clinical Trials GLP Analytics-GLP NMR Metabolite & Impurity Synthesis

1 www.selcia.comwwww.selcia.selcia.com Selcia Radiolabelling

Carbon-14 is a natural isotope of carbon and its decay is the basis of radiocarbon dating. When produced artificially it provides an essential tool to determine the behaviour and fate of chemical compounds in organisms and the environment.

Incorporation of 14C into a molecule of interest If the radiolabel is in a metabolically stable creates a radiotracer that is chemically and position it remains associated with the compound biologically identical to its non-labelled counterpart. and its metabolites.

A 14C-radiolabel allows easy detection with high The synthesis of a radiolabelled compound sensitivity in complex biological matrices, due is a good investment especially when taking to the low natural background of carbon-14. into account the relative ease of analytical This enables monitoring of both location and work and the quality, reliability and suitability speed of processes. of the data generated for regulatory submissions.

2 selcia.com/radiolabelling 3 Selcia RadiolabellingRadiolabelling

Typical studies using 14C-radiotracers

Penetration Radiotracers facilitate measurement of percutaneous penetration across membranes (i.e. skin) and may be used Metabolism in a diffusion cell, such as a Franz diffusion cell, to allow Metabolism studies using radiotracers can be performed determination of dermal absorption and to provide data in vitro and in vivo and require the radiolabel to be in on skin penetration for risk assessments. a stable position within the test molecule. Metabolism studies require sensitive detection, efficient separation, and accurate quantitation as well as isolation of the parent and the metabolites in biological matrices.

Absorption Various routes of administration (e.g. intravenous, oral, etc..) of a radiotracer allow monitoring of appearance Excretion and disappearance of the test substance in the blood or Without a radiolabelled compound an excretion mass plasma. Simultaneous dosing by oral and IV provides a balance study is extremely difficult. Formal mass balance measure of absolute bio availability. studies are required for all New Chemical Entities as part of a New Drug Application.

Distribution Radiotracers are invaluable for tissue distribution studies Environmental fate facilitating detection, separation, measurement and Radiotracers are instrumental in the study of the fate isolation of the test substance. When used in combination of substances in the environment. Such studies are a with mass spectroscopy techniques radiotracers help regulatory requirement for product registration, in particular determine if a peak is associated with either the parent or for pesticides. By studying metabolism, degradation and a metabolite. Accumulation can be measured after repeat dissipation of a chemical, its persistence in the environment doses. Quantitative Whole Body Autoradiography (QWBA) can be predicted. With the help of radiotracer the rate of is especially useful for tissue distribution studies. degradation of parent and metabolites in water, soil and other natural matrices can be measured accurately.

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14C Custom Radiolabelling Selcia is the leading independent carbon-14 radiolabelling company

Our independent position and established relationship with leading CROs gives you maximum flexibility when choosing the appropriate contract laboratory for your 14C study

14C Radiosynthesis including de novo Safe and fast shipment worldwide to development of synthetic routes multiple destinations Synthesis of radiolabelled API to GMP QC with full characterisation (HPLC, NMR, for clinical trials MS). Gravimetric analysis for specific activity, Chiral separation and salt formation radiochemical purity by HPLC. Chemical purity and structural identity by HPLC and NMR Rapid re-purification service and LC-MS Storage and stock management of GLP certification for regulatory submission radiolabelled compounds Long term storage at +4°C, -15 °C or -80 °C.

All our compounds are delivered with an industry leading data package, including full spectroscopic analysis and experimental documentation. A Certificate of Analysis specifying chemical as well as radiochemical purity by NMR, MS, radio-HPLC and radio-TLC accompanies each compound. The quality and detail of the data given in the data package make it suitable for regulatory submission.

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Carbon-14: The isotope of choice Characteristics of carbon-14 in the life science industry and 14C-radiotracers Low natural background Natural abundance: 1.18 x 10-12 in atmosphere Universal quantification No standard required Even minute amounts of radioactive Identity of analyte is not required for quantitation Rate of decay is constant and independent of pressure, material can be detected easily, temperature, pH and of chemical structure which makes it ideal for use in tracing the movements of highly

diluted substances in complex Carbon-14, the isotope of choice in the life science industry matrices, such as the environment Biologically equivalent to carbon-12. Introduction of 14C label does not change physico- or living organisms chemical properties or biological activity of molecules No shielding necessary; max. penetration in air: 20cm High detection sensitivity in biological matrices by LSC Long half life: 5730±40 years / No correction for decay necessary

14C is introduced into a defined position within the molecule through

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Selcia Synthetic Expertise A testimony to Selcia’s synthetic expertise

At Selcia we are at the forefront of technological Carbon-14 labelling of organic molecules requires “First Total Synthesis of Florian Viton, [a] Cyrille Landreau, [b] David development in our sector. Constantly reviewing total synthesis of the target structure from a basic 14 Rustidge, [b] Fabien Robert, [a] Gary Williamson, our skill portfolio we apply the latest techniques single carbon precursor such as [14C]cyanide or C-Labelled Procyanidin B2” [a] and Denis Barron[a] A Milestone Toward Understanding Cocoa in and purification such [14C]carbonate. The synthetic expertise of Selcia Polyphenol Metabolism as microwave-assisted reactions and chiral scientists enables us to prepare almost any [a] Nestlé Research Center, preparative HPLC to achieve the best possible chemical compound including complex P. O. Box 44, 1000 Lausanne 26, Switzerland results for our clients. natural products. [b] Selcia Ltd., Fyfield Business & Research Park, European Journal of Organic , 2008, Fyfield Road, Ongar, Essex, CM5 0GS, UK 36, 6069-6078

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GLP Analytical Services and Metabolite Synthesis

Our analytical staff have a strong background Selcia can act as Study Director or Principal supporting regulatory studies acquired over Investigator to support your GLP many years in the life science industry regulatory study

We provide specialist analytical services tailored We specialise in mass spectrometry to individual needs to support regulatory studies and particularly NMR spectroscopy. and submissions. Instrumentation in our GLP laboratory includes: GLP NMR for structural confirmation and Our analysts work closely with our comparison with standards and full synthetic chemists to resolve the High Field NMR, 300MHz, 400MHz and 500MHz spectrometers interpretation and structural elucidation chemical structures of unidentified GLP analysis including mass spectrometry process impurities, metabolites or Flow and capillary flow LC-NMR and infra-red spectroscopy degradation products. Our analytical 2H, 13C, 15N, 19F and 31P probes Custom synthesis of metabolites, impurities department is accredited by the MHRA Mass Spectrometry LC-MS/MS and reference standards and we are a full member of the Good Analytical and Preparative HPLC Custom synthesis of compounds labelled with Laboratory Practice program in the UK FT-IR stable isotopes such as Carbon-13, and specialise in GLP NMR to support Nitrogen-15, Deuterium regulatory submissions. All custom prepared products can be provided with a GLP certificate of analysis Non-routine analytical studies including structural elucidation and impurity profiling at microgram levels using capillary NMR

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fo r S e lc ia R a Working with Selcia d io la b e ll in g Credibility Communication P ro je c We have a tradition of applying scientific The dialogue begins from the moment we receive t excellence to satisfy the stringent quality criteria your enquiry. We take time to discuss your Quotation of our clients. We fully understand the importance requirements to ensure the radiolabelled product Request of delivering a high quality test substance on time will be of suitable specification for its intended use. CDA Agreed for the study. The success of your study is our Where necessary we can advise on label position, primary concern. quantity, specific activity, synthesis route and other factors as part of the quotation process and at no Personal service extra charge. Once we have agreed the proposal Consultancy and the project is underway, you will be able to on Isotope & Our business is built on fostering long-term communicate directly with the project scientist Label Position Quotation relationships with our clients. We work hard to to follow progress. Our scientists report get to know our customers and the specific pro-actively on the status of your project with a Confidentiality requirements of their business. We are in favour minimum of bi-weekly written progress reports Assured of face-to-face meetings and regular visits to help with further updates as soon as significant Route sustain these ongoing relationships. Equally we developments arise. Contract Development encourage clients to see our facility first hand. / Specification Informal visits are as welcome as audits. (Unlabelled) We strive to create transparent and open business & Delivery Time relationships built on trust to meet clear objectives. Agreed

Full QC Analysis & Radiosynthesis Data Package

Shipment, Stability Study & Storage

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Working with Selcia At Selcia we work to a rigorous quality program designed to ensure that the specifications of our products and services meet the client’s requirements in all respects

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Radioactive units Useful units and conversion for 14C radiotracer work

Characterisation of 14C labelled compounds Radioactive decay: 1Bq = 1 disintegration per second (SI unit)

Primary measure of the quality of a radiolabelled In order to use the radiochemical in quantitative Old unit, still in use: 1Ci = 3.7 x 1010 disintegrations per second compound is the “radiochemical purity”, which experiments its specific activity, i.e. the is determined by HPLC in conjunction with a radioactivity per mass unit has to be determined. Conversion: 1Ci = 3.7 x 1010Bq or 1mCi = 37MBq radiodetecor (liquid scintillation flow counter). This is typically achieved by gravimetric analysis, The radiochemical purity is directly obtained by the direct measurement of the radioactivity in a Specific activity (SA): Radioactivity per mass unit integrating the radio-HPLC trace. No calibration known amount of radioactive sample. Most commonly used in 14C tracer work: against a standard is required. The result obtained is MBq/mg or µCi/mg. 1µCi/mg or 1MBq/mg

Chemical purity is determined by conventional This can be converted into molar specific activity In it is common to use molar specific activity: HLPC methods and NMR. by the formula shown in the box on the next page. mCi/mmol or MBq/mmol

The max. specific activity for 14C , i.e. 1 atom of 14C per molecule is 62.4mCi/mmol (theoretical maximum) in practice max. 50-60mCi/mmol

Conversion: 1mCi/mmol = 1mCi/mg x MW (MW = molecular weight)

14 Example: Ba CO3 (MW = 199.10) 55mCi/mmol is equivalent to 0.276mCi/mg or 10.2MBq/mg

Formula to calculate the adjusted molecular weight of a radiolabelled compound from its specific activity:

-1 MW MW SA [mCi mmol ] x 2 labelled = unlabelled + 62.4mCi mmol-1

or in MBq

-1 MW MW SA [MBq mmol ] x 2 labelled = unlabelled + 2309MBq mmol-1

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Carbon-14 facts

Carbon-14 is a cosmagenic isotope 1n + 14N -> 14C + 1p produced in nature through cosmic Natural abundance = 1.18 x 10-12. Approximately 1 atom in 1 trillion ray bombardment of nitrogen at high altitudes in the atmosphere Carbon-14 timeline

14 C -> 14 N + e- + v Carbon-14 first postulated Carbon-14 decay 6 7 e Low energy ß—emitter (Emax=156keV) 1934 by Franz Kurie Half life T1/2 = 5730±40 years

Maximum molar specific activity SAmax=62.4mCi/mmol First detected and isolated by 1940 Martin Kamen and Sam Ruben

First studies with 14C-radiotracer: Incorporation of 14CO into plants 1942 2 14C ~5,730 years 14N

Libby states radioactive 14C 1947 is found in all living creatures - Libby et al. receive Nobel e 1960 prize for carbon dating

Calvin wins Nobel prize for 1961 work on photosynthesis

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AboutAbout SSelciaelcia

A leading worldwideworldwide pproviderrovideider ooff iintegratedntegrated ddrugrug discoverydiscovery andand 1414C ccustomustom rradiolabellingaadiolabellingdiolabelling sservices.ervices. SelciaSelcia is a pprivatelyrivately hheldeld ccompanyompany opeoperatingratin ffromrom state-of–tstate-of–the-arthe-art labolaboratories near London, United KKingdom.

Integrated Drug Discovery 1414C Custom Radiosynthesis Macrocycles & Natural Products 1414C GMP API for Clinical Trials MMedicinaledicinal CChemistryhemistry GLP Analytics-GLP NMR PPIase Screening Metabolite Synthesis drug discovery

Selcia Ltd. Fyfield Business & Research Park, Fyfield Road, Ongar, Essex. CM5 0GS United Kingdom

+ 44 (0) 12127777 363677 000000 [email protected]@selcia.com