The Value of Digital PhotographyBy Louise Rutherford and Dean Hatt

Agenda

–Introduction–Case Study A–Case Study B–Case Study C–Case Study D–Case Study E–Conclusions–Questions

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Introduction

– Digital photography was introduced into the Dispensary in 2008 to primarily assess potential colour changes on GLP DMSO stability studies

– Over the last 6 years we have managed to capture digital images on over 150 stability studies

– Approximately 20% of these have demonstrated a colour change over time

– The set up and use of digital photography on stability studies has previously been presented to the CHAOTIC group

– The use of digital photography has now been extended beyond stability assessments

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Case Study A

– Compound A was tested within Pharmaceutical Development on a small scale and shown to have high aqueous solubility

– A formulation in saline was required for continuous infusion to support in-vivo studies

– Dispensary were responsible for the development of in-vitro formulations in DMSO/Water/hERG PSS

– Initiated a solubility assessment on an early batch of compound

Solubility Assessment

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Compound A – Early BatchSolubility in Water

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Compound A – Formulations in Saline

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Compound A. Batch Colour Differences

– Early Batch

– New Batch

Compound A

– To summarise:

– Problem 1 – Formulation changing form over time producing ‘worm cast’

– Problem 2 – Dramatic colour difference between batches

– Problem 3 – Extraordinary differences in formulations of the SAME batch prepared in the same conditions but using compound from different containers

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Multiple Problems

Pharmaceutical Development Container – New Batch

Dispensary Container – New Batch

Case Study A

– Pictures were sent to the project team which immediately allowed escalation of the issue and meetings were set up to discuss.

– Lab investigations started in collaboration between SA and Pharmaceutical Development

– Likelihood this is related to the formation of a trihydrate

– Continuous infusion studies were delayed as a result of the finding

– Formulation work is ongoing

Conclusions – Where Are We Now

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Case Study B

– DRF’s had successfully been completed on early batches

– Standard suspension in 1% MC at reasonably low concentration – up to 10 mg/mL.

– GLP batch was received and stability study was initiated

Oral Stability Study

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Compound B

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‘Gumming’

Compound B

– Formulation was stable at up to 1 mg/mL in 1% MC

– Formulation not suitable at 10 mg/mL

– In-vivo studies potentially required up to 10 mg/mL

– SA dispensary conducted a vehicle assessment study

– Use of surfactants and intermittent sonnication/stirring were identified as the likely route to take

– The addition of 0.2% SLS was selected with the following result........

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Unsuitable vehicle

Case Study B

– Digital photography was utilised to demonstrate the formulation issue to both SA project team member and Pharmaceutical Development

– Agreed that the formulation was unsuitable

– This was taken forward to successful stability study with this new vehicle

– Both stable and homogenous

– This was successfully used in GLP studies

– This has also been used with a new salt form of the same drug and this information was also used in developing a further formulation for MNT study

Conclusion

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Case Study C

– Stability studies in Water, hERG PSS and an aqueous buffer were successfully conducted up to 100 mg/mL– Solubility assessment demonstrated up to approx 400 mg/mL in all

three vehicles

– Conducted on an early DRF batch that was made to GxP

– All GLP studies scheduled to start at CRO including one month nebulised inhalation studies

– A haemolysis assay was conducted in house using the new GLP batch

Starting GLP studies

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Compound C

5 mg/mL in Buffer Vehicle

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Compound C 12 mg/mL in Buffer Vehicle

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Compound C

New Batch v Old Batch New Batch – Water V DMSO

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Compound C

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50 mg/mL in buffer vehicle

– Dark Deposits on glass

Case Study C

– Digital photography was able to demonstrate a clear difference in solubility of the new batch

– < 5 mg/mL compared with approx 400 mg/mL

– Dramatic colour difference between the two batches

– Pictures clearly showed the appearance of a dark red substance throughout the formulations

– Meetings were held between SA and Pharmaceutical Development

– Samples of all formulations sent to Pharmaceutical Development for analysis

– Pharm Dev could not identify the dark red substance but analysis results were close to nominal

– CRO studies were postponed to allow clean up and formulation re-work to be performed

– This was high profile as it was one of the first outsourced packages using the new SA strategy.

Conclusions

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Case Study D

– Carcinogenicity study at CRO about to begin

– Formulation in 0.5% MC

– Compound was a sodium salt

– pH was at the top end of the acceptable range pH 12 at 200 mg/mL

– Potential reverse solubility but the dark colours made this difficult to assess

– Trial formulations at the CRO demonstrated poor physical stability and crystallisation of formulations which failed to re-suspend

– SA Dispensary conducted a further stability study looking at the physical stability and how we could advise the CRO on how they could prepare it

Physical Instability

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Compound D

200 mg/mL Day 0 20 mg/mL Day 0

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Reverse Solubility

Compound D

Day 7 200 mg/mL not stirred Day 7 60 mg/mL not stirred

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Colour Differences

Compound D

20 mg/mL 60 mg/mL

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Day 15 stirred v non stirred

Compound D

200 mg/mL

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Stirred v Non Stirred

Case Study D

– Digital photography was used to demonstrate a time course of physical stability at a range of concentrations and the effect of continuous stirring

– Crystallisation was not seen on the scale performed in this stability study

– Project team were happy for us to make recommendations as to the frequency and scale / detail of preparation to allow consistent formulations on study

– At 200 mg/mL the recommendation was to prepare the day prior to dosing and store without continuous stirring

– At concentrations between 0.1 and 60 mg/mL the recommendation was to prepare at least 3 days prior to dosing to ensure that a suspension was dose

Conclusions

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Case Study E

– Current formulation contained 2-hydroxypropyl beta cyclodextrin

– FDA rejected CAC proposal for use in a 2 year carc study due to link with cyclodextrins and pancreatic tumours in rats

– Had low and variable exposure

– SA conducted a vehicle assessment study

– Vehicle selected was 50:50 (v/v) PEG 400: Saline

– Formulation a solution

– Solution at high levels potentially required heat to maintain

– Stability study conducted to assess chemical and physical stability

Temperature Effect on Oral Stability Study

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Compound E

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Temperature Related Colour Change

– 10 mg/mL Day 0 – 10 mg/mL Day 7 37°C

– 10 mg/mL Day 15 37°C

– 10 mg/mL Day 15 4°C

Compound E

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– Day 0 100 mg/mL – Day 7 100 mg/mL 37°C

– Day 15 100 mg/mL 37°C

– Day 15 100 mg/mL 4°C

Stability Results

Day 0 Day 7 (approx 37°C)

Day 15 (approx 37°C)

Day 15 (approx 4°C)

Determined Sample Concentration (mg/mL)

9.041 8.72 8.711 9.171

% of Nominal Concentration 90.4 87.2 87.1 91.7

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– 10 mg/mL Stability Results

Day 0 Day 7 (approx 37°C)

Day 15 (approx 37°C)

Day 15 (approx 4°C)

Determined Sample Concentration (mg/mL)

94.61 98.9 98.51 98.71

% of Nominal Concentration 94.6 98.9 98.5 98.7

– 200 mg/mL Stability Results

Compound E

Day 0 0.1 mg/mL Day 15 0.1 mg/mL – 37°C

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Day 0 Day 7 (approx 37°C)

Day 15 (approx 37°C)

Day 15 (approx 4°C)

Determined Sample Concentration (mg/mL)

0.11 0.0943 0.08321 0.09861

% of Nominal Concentration

100 94.3 83.2 98.6

Case Study E

– Formulation provided increased and non variable exposure in in-vivo studies

– Formulations were chemically stable for up to 15 days at 0.1-100 mg/mL when stored at approximately 4°C

– At 100 mg/mL the formulation failed to stay in solution at both tested temperatures, but was re-solubilised when minimal heat applied and/or maintained

– Formulations were chemically stable for up to 15 days at 10-100 mg/mL when stored at approximately 37°C but unstable at 0.1 mg/mL at this storage condition

– Digital photography was used to assess colour change throughout all phases

– Colour changes at 10 and 100 mg/mL and chemical instability at 0.1 mg/mL following storage at 37°C, preclude maintenance of these solutions at this temperature

– Solutions at 100 mg/mL to be stored at 4°C and re-solubilised for dosing period only

Conclusions

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Overall Conclusions

– Digital photography can be used beyond the original remit of assessing instability

– It is a useful tool for assessing:

– Batch variation

– Insolubility

– Physical instability

– It can therefore aid in:

– Proactive formulation assessment

– Reactive formulation issues

– Fast, non subjective communication of formulation issues to the relevant people

– This is of greater importance with the SA strategy of outsourcing GLP studies where formulations are prepared at a greater scale and on batches manufactured using different synthetic routes

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Questions?