Analysis of Excipients in Inhaled Dose Formulations Chaotic

Analysis of Excipients in Inhaled Dose Formulations Chaotic Meeting October 2014

Agenda • What are excipients? • Why look? • What was required? • How we did it • Results • Conclusions • Future Work Analysis of Excipients in inhaled Dose Formulations Chaotic Meeting October 2014

What are Excipients? An excipient is a natural or synthetic substance formulated alongside the active ingredient of a medication, included for the purpose of bulking-up formulations that contain potent active ingredients. They are often referred to as “bulking agents”, “fillers” or “diluents” Excipients can be useful in the manufacturing process, to aid in the handling of the active substance concerned such as facilitate powder flowability or non stick properties. The performance of a formulation may be engineered by manipulating the physical and chemical properties of excipients and coatings. Excipients help define the bioavailability, release profile and stability of a formulation. The selection of appropriate excipients can also be determined on the route of administration and dosage form. Analysis of Excipients in Inhaled Dose Formulations Chaotic Meeting October 2014

Why look? • Excipients were considered at one time to be “inactive” ingredients, however they are now thought to be “a key determinant of dosage form performance” [1] • Pharmaceutical regulations and standards require that all ingredients in drugs, as well as their decomposition products, be identified and shown to be safe. • [1] Bhattacharyya, Lokesh; Schuber, Stefan; Sheehan, Catherine; William, Roger (2006). "Excipients: Background/Introduction". In Katdare, Ashok; Chaubal, Mahesh. Excipient Development for Pharmaceutical, Biotechnology, and Drug Delivery Systems Analysis of Excipients in Inhaled Dose Formulations Chaotic Meeting October 2014

What was Required? • At GSK pre-existing excipients Poly Lactic-co-Glycolic Acid grade 2 A(PLGA), Hydroxypropylmethylcellulose acetyl succinate grade LF(HPMC-AS), Trehalose and Tri-Leucine, currently unapproved for inhalation use, are being investigated for the inhalation programme. • An investigative study (R 30894 N) was performed dosing all 4 excipients to male Rats for 14 days, to assess any progression/regression of toxicity. • Dose formulation analysis methods were required to support the study. – External pre-existing methods were made available, however availability of equipment and time dictated redevelopment in-house. – Trehalose & Tri-Leucine were dosed at a 1: 1 ratio, so it was felt that only Trehalose would be analysed. Analysis of Excipients in Inhaled Dose Formulations Chaotic Meeting October 2014

How we did it – PLGA & HPMC-AS • Materials – Waters Styragel HR 0. 5 THF, 4. 6 x 300 mm – Tetrahydrofuran (THF) – Agilent 1100 with UV Detection 218 nm – Glass fibre filters (37 mm circles) • Standards & Samples – Diluent (THF), added to sample filters and blank filters. – Calibration standards prepared over the range 50 to 1000 µg/m. L, 1/x linear regression. – Quality Controls prepared at 3 levels within the range. • Chromatography – Isocratic, 0. 5 m. L/min, 100% THF – 10µL injection, 15µL injection HPMC-AS – 10 min Run Time – Ambient column temperature Analysis of Excipients in Inhaled Dose Formulations Chaotic Meeting October 2014

PLGA 50 µg/m. L LLQ Analysis of Excipients in Dose Concentration Formulations - Chaotic

HPMC-AS 100 µg/m. L LLQ Analysis of Excipients in Dose Concentration Formulations - Chaotic

How we did it – Trehalose • Materials – Waters BEH Amide 1. 7 µm, 50 x 2. 1 mm – Water, Acetonitrile – Waters Acquity UPLC with Charged Aerosol Detection (CAD) – Glass fibre filters (37 mm circles) • Standards & Samples – Diluent (20: 80 v/v acetonitrile/water), added to sample filters and blank filters. – Calibration standards prepared over the range 40 to 1000 µg/m. L, 1/x quadratic regression. – Quality Controls prepared at 3 levels within the range. • Chromatography – Isocratic, 1 m. L/min, 20: 80 v/v water/acetonitrile – 1µL injection – 1. 5 min Run Time – 50ºC column temperature Analysis of Excipients in Inhaled Dose Formulations Chaotic Meeting October 2014

Trehalose 40 µg/m. L LLQ Analysis of Excipients in Dose Concentration Formulations - Chaotic

Results PLGA • Validated successfully over the range 50 – 1000 µg/m. L in GFA filters and THF solutions at 3 QC levels. • 20% acceptance criteria from the Nominals. • GFC filters did not validate at 50 µg/m. L • 1/x weighted linear regression Analysis of Excipients in Inhaked Dose Concentration Formulations Chaotic Meeting October 2014

Results PLGA QC 50 SOLN QC GF/A 50 QC GF/C 50 Rep 1 51. 115 43. 327 32. 955 Rep 2 51. 261 39. 254 34. 321 Rep 3 51. 150 39. 672 34. 155 Mean 51. 175 40. 751 33. 810 % Bias -2. 351 18. 498 32. 379 QC 200 SOLN QC GF/A 200 QC GF/C 200 Rep 1 212. 486 177. 876 166. 744 Rep 2 198. 700 179. 875 167. 112 Rep 3 199. 296 186. 329 167. 926 Mean 203. 494 181. 360 167. 261 % Bias -1. 747 9. 320 16. 370 QC 1000 SOLN QC GF/A 1000 QC GF/C 1000 Rep 1 955. 499 965. 524 951. 159 Rep 2 956. 349 972. 941 963. 886 Rep 3 964. 640 977. 502 961. 510 Mean 958. 829 971. 989 958. 852 % Bias 4. 117 2. 801 4. 115 Analysis of Excipients in Dose Concentration Formulations Chaotic Meeting October 2014

Results HPMC-AS • Did not validate over the range 50 – 1000 µg/m. L, LLQ raised to 100 µg/m. L. • 50 µg/m. L QC’s did not validate • Assumed ok from 100 µg/m. L in GFA filters and THF solutions • 100 µg/m. L replicates included in the first study run • 15% acceptance criteria from the Nominals. • GFC filters not included • 1/x weighted linear regression Analysis of Excipients in Dose Concentration Formulations Chaotic

Results HPMC-AS QC 50 SOLN QC GF/A 50 Rep 1 23. 400 19. 700 Rep 2 23. 100 20. 100 Rep 3 23. 500 19. 700 Mean 23. 333 19. 833 % Bias 53. 333 60. 333 QC 500 SOLN QC GF/A 500 Rep 1 483. 000 465. 300 Rep 2 483. 900 466. 300 Rep 3 483. 500 468. 300 Mean 483. 467 466. 633 % Bias 3. 307 6. 673 QC 1000 SOLN QC GF/A 1000 Rep 1 1065. 600 947. 500 Rep 2 1064. 100 945. 600 Rep 3 1070. 600 948. 300 Mean 1066. 767 947. 133 % Bias -6. 677 5. 287 Analysis of Excipients in Dose Concentration Formulations Chaotic Meeting October 2014

Results Trehalose • Did not validate over the range 20 – 1000 µg/m. L, LLQ raised to 40 µg/m. L. • Assumed ok from 40 µg/m. L in GFA filters and 20/80 acetonitrile/water solutions • 40 µg/m. L replicates included in the first study run • 15% acceptance criteria from the Nominals. • GFC filters not included • 1/x weighted quadratic regression Analysis of Excipients in Dose Concentration Formulations Chaotic Meeting October 2014

Results Trehalose QC 20 SOLN QC GF/A 20 Rep 1 17. 200 20. 900 Rep 2 14. 200 18. 000 Rep 3 15. 700 17. 300 Mean 15. 700 18. 733 % Bias 21. 500 6. 333 QC 200 SOLN QC GF/A 200 Rep 1 189. 600 197. 100 Rep 2 190. 800 185. 900 Rep 3 197. 100 191. 100 Mean 192. 500 191. 367 % Bias 3. 750 4. 317 QC 1000 SOLN QC GF/A 1000 Rep 1 993. 200 941. 500 Rep 2 964. 500 1025. 900 Rep 3 1018. 800 930. 000 Mean 992. 167 965. 800 % Bias 0. 783 3. 420 Analysis of Excipients in Dose Concentration Formulations - Chaotic

Conclusions • Analytical methods were – Fit for purpose for this investigative study • PLGA showed a favourable safety profile over 14 days in rats exposed to doses up to 36 mg/kg/day, however alongside the test article it showed instability issues. • Trehalose: Tri-Leucine showed a favourable safety profile over 14 days in rats exposed to doses up to 66 mg/kg/day and considered suitable for future progression. • HPMC-AS showed an unfavourable lung pathology profile at the proposed clinical doses. The data did not preclude use at lower clinical doses. • Safety Assessment now only use GFA filters. Analysis of Excipients in Inhaled Dose Formulations Chaotic Meeting October 2014

Future Work Studies PLGA maybe considered acceptable for use alongside other test articles HPMC-AS further safety work would be required Methods Re-visit PLGA & HPMC-AS method chromatography, using a more readily available and user friendly solvent THF is highly volatile and pungent Explore the possibility of combination assays Safety Assessment standardised on GFA filters Analysis of Excipients in Inhaled Dose Formulations Chaotic Meeting October 2014

Thanks…. Paul Giffen Kay Rush Dean Hatt Teresa Fuller Louise Kay Aida Merchant Analysis of Excipients in Dose Concentration Formulations Chaotic Meeting October 2014

Questions? Analysis of Excipients in Dose Concentration Formulations Chaotic Meeting October 2014