Therapeutics Ltd Research Articles

VIEWS and NEWS A celery gene may protect roses from disease • 15th European Congress on Biotechnology • Environmental Microbiology and Biotechnology Conference 2012 • FDA approved first cell therapy against wrinkles • Individualization of cancer treatment using personalized tumorgrafts • Mission Therapeutics Ltd, a new British spin-out company, raised £6 million • Science Multimedia Center

VIEWS and NEWS A celery gene may protect roses from disease • 15th European Congress on Biotechnology • Environmental Microbiology and Biotechnology Conference 2012 • FDA approved first cell therapy against wrinkles • Individualization of cancer treatment using personalized tumorgrafts • Mission Therapeutics Ltd, a new British spin-out company, raised £6 million • Science Multimedia Center

BKT140 Is a Novel CXCR4 Antagonist with Stem Cell Mobilization and Antimyeloma Effects: An Open-Label First Human Trial In Patients with Multiple Myeloma Undergoing Stem Cell Mobilization for Autologous Transplantation

AbstractAbstract 2260 Background:BKT140 is a high affinity CXCR4 inhibitor with an extended K off-rate. Pre-clinical studies in animal models with BKT140 showed a robust mobilization of white blood cells (WBC) and hematopoietic stem cells (HSC). Furthermore, BKT140 also showed a direct anti-tumor effect against human-derived multiple myeloma (MM), lymphoma and primary leukemia cells and cell lines in vitro and in vivo, causing significant apoptosis. Aims:To assess BKT140 toxicity (primary endpoints), the mobilization capacity of CD34+ hematopoietic progenitors and CD138 MM cells, and pharmacokinetic (PK) and pharmacodynamic (PD) (secondary endpoints). Methods:16 MM patients in first CR/PR were included in a phase I/IIa study, in which escalating doses of BKT140 (30, 100,300,900 μg/kg) were administered together with a high-dose cyclophosphamide (Cy) (2 g/m2) and G-CSF (5 μg/Kg) for stem cell mobilization. G-CSF was started on day 5 post Cy and BKT140 was injected subcutaneously (SC) once on day 10. Toxicity, PK, and mobilization capacity (assessed by serial measurements of number of WBC and CD34+ and CD138+ cells) were measured pre- and post BKT140 administration. Results:BKT140 was well tolerated at all doses and none of the patients developed grade II-IV toxicity. BKT140 was rapidly absorbed with no observed lag time, with peak plasma concentrations occurring 0.5h after administration. Clearance was rapid, with a median terminal half-life of 0.69h. BKT140 administration resulted in a significant dose-dependent increase in the number of peripheral blood neutrophils, monocytes, lymphocytes, and CD34+ cells compared to the G-CSF/Cy individual patient baseline. The maximum increase in the number of WBC from baseline was observed within 8h following BKT140 injection, 2.5-, and 3.0-, 4.1- and 4.8-fold, for the 4 BKT140 doses, respectively. Furthermore, BKT140 administration resulted in a significant increase in the mean absolute PB CD34+ cells mobilized (6.6, 7.5, 11.2 and 20.6 ×106/kg) for the 4 BKT140 administered doses, respectively. Moreover, the number of aphaeresis was reduced from 2.25 procedures at the first two BKT140 doses to 1.25 and 1 aphaeresis at the highest BKT140 doses, respectively. An increase in the number of CD138+ cells was observed in 6 out of 6 pts that had CD138+ cells in their blood and were treated with lower doses of BKT140 (30 and 100 μg/kg). Interestingly, in pts that were treated with the highest doses of BKT140 (300 and 900 μg/kg) a reduced number of CD138+ cells was observed in 3 out of 7 pts that had CD138+ cells in their blood, whereas in 4 pts, an increase in the number of CD138+ cells was shown. Three pts who did not have CD138+ cells in their blood were not affected by BKT140. The BKT140 mobilized grafts were used for AutoSCT following 200 mg/m2 melphalan conditioning. Pts received an average of 5.3×106 CD34+ cells/kg. All transplanted pts rapidly engrafted (n=15). The median day for neutrophil (>500/mm3) and platelet (>20,000/mm3, >50,000/mm3,) was on day 11 (range, 0–13), day 11 (range, 0–14), and day 14 (range, 0–23), respectively. Conclusions:The current data suggests that BKT140 can safely be added to G-CSF-based harvesting regimens, can increase CD34+ cell mobilization and reduce the number of collection days. Furthermore, due to its ability to release MM cells from the bone marrow and stimulate their cell death, additional studies are warranted to further evaluate the effect of BKT140 as an anti-MM agent. Disclosures:Nagler: Biokine Therapeutics Ltd: Consultancy. Abraham: Biokine Therapeutics Ltd: Employment, Equity Ownership, Patents & Royalties. Wald: Biokine Therapeutics Ltd: Employment. Shaw: Biokine Therapeutics Ltd: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees. Eizenberg: Biokine Therapeutics Ltd: Employment, Equity Ownership, Patents & Royalties. Peled: Biokine Therapeutics Ltd: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties, Research Funding.

Adaptation and Validation of the Plasma Inhibitory Activity (PIA) Assay to Detect Inhibition of Aurora, ABL and FLT3 Kinases by AT9283 In Children and Adolescents with Leukaemia

AbstractAbstract 1818Non-invasive assessment of biomarker modulation is critically important when conducting early phase trials of targeted therapeutics, particularly in children. Measuring target modulation using primary leukaemic blasts is challenging, since obtaining bone marrow is invasive and invariably requires a general anaesthetic in children. Furthermore, in many cases only a limited number of circulating blasts can be detected in peripheral blood and these may be rapidly cleared upon commencement of effective therapy. For these two reasons, pharmacodynamic (PD) assays using surrogate biological material can be invaluable. The plasma inhibitory activity (PIA) assay was initially developed for FLT3 inhibitors and is currently used in clinical trials to assess ex vivo inhibition of FLT3 kinase and guide optimal dosing (Levis M et al, Blood 2006). Briefly, the assay involves incubating reference leukaemic cell lines in plasma from patients treated with FLT3 inhibitors then assessing the degree of FLT3 inhibition by western blotting. Inhibition of FLT3 in leukemia cell lines has been shown to correlate with inhibition in primary leukaemic blasts. Aurora kinase inhibitors are emerging as promising new agents with activity in leukaemia (Moore AS et al, Leukemia 2010). AT9283 is a novel Aurora kinase inhibitor with activity against the secondary kinase targets FLT3 and ABL (Howard S et al, J Med Chem 2009). We therefore hypothesised that the PIA assay would be applicable as an ex vivo PD assay to simultaneously detect inhibition of these target kinases in leukaemic cell lines: Aurora with FLT3 and Aurora with ABL. Furthermore, we hypothesised that the assay could be adapted to paediatric patients, where limited volumes of blood are available for PD studies. Here we report the validation of the PIA assay for use with AT9283 using leukaemic cell lines incubated in human plasma spiked with clinically relevant concentrations of AT9283 known to inhibit Aurora kinase in primary leukaemic blasts. The PIA assay proved to be a robust means of semi-quantitatively detecting concentration-dependent inhibition of Aurora kinase in the FLT3-ITD positive AML cell line MOLM-13 and the CML cell line K-562. The PIA assay was also effective at qualitatively detecting the inhibition of FLT3 signaling in MOLM-13 cells and ABL signaling in K-562 cells. Of importance to paediatric patients, the assay was successfully performed using 0.5 mL of plasma, half the volume described in the original PIA assay for FLT3 inhibitors. It will be used to assess target kinase modulation in a phase I trial of AT9283 in children and adolescents with relapsed and refractory acute leukaemia (EudraCT No. 2009–016952-36). In conclusion, the PIA assay is applicable not only to FLT3 inhibitors, but also Aurora kinase inhibitors and potentially, other multi-kinase inhibitors. By simultaneously detecting multiple kinase inhibition, the PIA assay may help delineate important mechanisms of action for novel anti-leukaemic drugs. Disclosures:Podesta:The Institute of Cancer Research: Employment, The Institute of Cancer Research (ICR) has a commercial interest in drug development programs (www.icr.ac.uk). Authors employed by ICR are subject to a Rewards to Inventors Scheme which may reward contributors to a program that is subsequently licensed. Squires:Astex Therapeutics Ltd: Employment. Linardopoulos: The Institute of Cancer Research: Employment, The Institute of Cancer Research (ICR) has a commercial interest in drug development programs (www.icr.ac.uk). Authors employed by ICR are subject to a Rewards to Inventors Scheme which may reward contributors to a program that is subsequently licensed. Pearson:The Institute of Cancer Research: Employment, The Institute of Cancer Research (ICR) has a commercial interest in drug development programs (www.icr.ac.uk). Authors employed by ICR are subject to a Rewards to Inventors Scheme which may reward contributors to a program that is subsequently licensed. Moore:The Institute of Cancer Research: Employment, The Institute of Cancer Research (ICR) has a commercial interest in drug development programs (www.icr.ac.uk). Authors employed by ICR are subject to a Rewards to Inventors Scheme which may reward contributors to a program that is subsequently licensed.

AT9283, a Small Molecule Multi-Targeted Kinase Inhibitor with Potent Activity Against Aurora Kinase and STAT3 In Combination with Lenalidomide Results In Synergistic Anti-Myeloma Activity

AbstractAbstract 2994Despite recent advances with new drugs such as bortezomib, thalidomide and lenalidomide, multiple myeloma (MM) remains an incurable disease. Used as single agents, these compounds have shown marked antitumor activity, but the number of patients with relapsed and refractory disease remains high. Combination of these agents with other classes of novel drugs would offer great promise to improve patient outcome. AT9283 (Astex therapeutics, Cambridge UK) is a multi-targeted kinase inhibitor that inhibits Aurora A (AURKA), Aurora B (AURKB) and Janus Kinase (JAKs). AURKA and AURKB expression has been correlated with genetic instability and cellular proliferation in MM; therefore, Aurora kinases represent an attractive therapeutic target in MM. In addition the JAK/STAT pathway plays an important role in the survival and proliferation of MM cells. Blocking this pathway may therefore be critical for the survival of MM cells. AT9283 decreased both phospho-Histone H3 and the phosphorylation of Aurora A at Thr 288 in Nocodazole treated cells, suggesting the dual activity of AT9283 against AURKA and AURKB. Importantly, besides Aurora kinase inhibition, we observed that AT9283 inhibited STAT3 tyrosine phosphorylation within 30 minutes of treatment. The effect of AT9283 on pSTAT3 inhibition was further investigated by using U3A cells stably expressing a luciferase reporter gene under the control of a STAT-dependent promoter. AT9283 inhibited STAT3-dependent luciferase activity with an EC50 of approximately 0.125 μ M. Consistent with AT9283 induced cytotoxicity, genetic depletion of STAT3, AURKA or AURKB showed growth inhibition of MM cells, suggesting that AT9283-induced inhibition of these molecules is in part the underlying mechanism of MM cell growth inhibition. In vivo data using a xenograft mouse model of human MM show that mice treated with AT9283 demonstrated slower tumor growth compared to the control group (p=0.018) and prolongation in median overall survival (32 days in treated group versus 18 days in control group; p < 0.0001) without adverse effects. We next evaluated the activity of AT9283 in combination with established MM drugs and strong synergistic effect was found when AT9283 was combined with lenalidomide (Selleck Chemicals LLC, TX, USA) (Combination Index < 0.9). We hypothesized that the synergistic effect of this combination is due to the fact that the two drugs target different pathways and different phases of the cell cycle, thus augmenting their individual anti-myeloma activity. We examined MM cell cytotoxicity of the combination by using AT9283 and lenalidomide at concentrations lower than their maximal cytotoxic concentrations. Increasing doses of AT9283 (0 -0.125 μ M) were added to lenalidomide (0-2μ M) and a significant decrease in viability (as measured by MTT and cell growth as determined by 3H-TdR at 48 h) was observed with combined therapy compared to either agent alone. A significant increase (55.7%) in early and late apoptosis occurred after 72 hours of exposure of cells to combined therapy with associated caspase-8 and PARP cleavage. Combination treatment resulted in downregulation of pSTAT3 and pERK following 4 hours of treatment. Considering the role that the BM microenvironment plays in growth and survival of MM cells, we examined whether the combination of low dose AT9283 plus lenalidomide induced MM cell death in the context of the BM microenvironment. MM.1S cells were cultured with or without BMSCs in the presence or absence of AT9283, lenalidomide or in combination regimen. Combined therapy inhibited 3H-TdR uptake of MM.1S cells cultured in the presence of BMSCs. Interestingly, we observed that AT9283 plus lenalidomide downregulated the expression of the p-STAT3 and p-ERK when MM.1S cells were cultured with BMSCs, highlighting the role of this drug combination in overcoming the protective effect of BMSCs. These results provide the rationale for the clinical evaluation of AT9283 in combination with lenalidomide in MM patients. Disclosures:Squires:3Astex Therapeutics Ltd: Employment. Anderson:MILLENNIUM: Consultancy; CELGENE: Consultancy; NOVARTIS: Consultancy; MERCK: Consultancy; ONYX: Consultancy; BMS: Consultancy. Raje:novartis: Consultancy; celgene: Consultancy; astra zeneca: Research Funding; acetylon: Research Funding.

The OPAL study: A phase II study to evaluate the efficacy, safety, and tolerability of tosedostat (CHR-2797) in elderly subjects with treatment refractory or relapsed acute myeloid leukemia.

TPS277 Background: Tosedostat (CHR-2797) is a novel oral aminopeptidase inhibitor. Aminopeptidases catalyze hydrolysis of terminal amino acids from (poly)peptides generated by proteasomal degradation. Inhibition of aminopeptidases leads to a deficiency of amino acids for protein synthesis. Malignant cells appear to be more dependent on protein cycling, so this pathway is a potential target for novel agents. In preclinical experiments, tosedostat exerts antineoplastic activity in solid tumor, leukemia and myeloma cell lines in vitro. A phase I-II study with tosedostat included 51 patients with acute myeloid leukemia (AML), and showed promising antileukemia effects and good tolerability. Thirty-five of these patients had relapsed or refractory AML. Complete response (CR) was defined as <5% blasts, partial response (PR) as 5-15% blasts. 11 of 35 had favorable responses (6 CRs, 5 PRs), an overall response of 31.4%. In follow-up, 5 of the 6 CRs were observed to result in 3 Complete Remissions, 1 CRp and 1 CRi. Longer median overall survival was noted for complete (334 days) and partial (361 days) responses compared to all patients (118 days) or nonresponders (97 days). This study formed the basis for the ongoing study. Methods: The OPAL study is a phase II study in 2 parts. Part A is a dose selection part in which 60 patients aged over 65 years with refractory or relapsed AML, whose first induction resulted in a complete remission for up to 12 months (Estey 1996), will be randomized to receive first salvage with tosedostat 120 mg daily for 6 months OR tosedostat 240 mg daily for 2 months followed by 120 mg for 4 months. A benefit-risk analysis will take place to determine the dose for Part B in which a further 130 patients will be treated with tosedostat. Responses will be assessed using IWG criteria (Cheson et al 2003). Results: Five patients were randomized by January 8, 2010. Updated enrollment numbers will be presented. Part B will commence in Q3/2010. Conclusions: Tosedostat is a promising agent for the treatment of refractory or relapsed elderly AML patients. The OPAL study will further evaluate the efficacy and safety of tosedostat in these patients. Author Disclosure Employment or Leadership Position Consultant or Advisory Role Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration Chroma Therapeutics Ltd. Chroma Therapeutics Ltd. Chroma Therapeutics Ltd.

Evaluation of length, volume and gray-scale histogram of the cervix as predictors of successful induction

Objective: The aim of this study was to evaluate whether the Bishop score, length, volume and gray-scale histogram of the cervix has a predictive value of assessing the rate of success in trial of induction. Methods: Forty-one nulliparous patients with its Bishop score six or less were enrolled for this prospective study. All were on prostaglandin (, Propess ; Controlled Therapeutics Ltd) pessary. Three-dimensional transvaginal ultrasound scans of the cervix were performed on the ACCUVIX XQ (Medison) to measure length, volume, and gray-scale histogram. Bishop score was determined by digital examination. The successful induction was defined as the ability to achieve the active phase of labor corresponding to a cervical dilatation of 4 cm within 12 hours of removing the pessary. The receiver operating characteristics (ROC) curves were also used to estimate an optimal cutoff point for the Bishop score, length, volume, and gray-scale histogram of the cervix. Logistic regression analysis was used for statistical analyses. Results: The overall successful rate of labor induction was 73.2% (30/41). Multiple logistic regression analyses demonstrated that the value of anterior lip histogram was significantly associated with the successful labor induction. ROC curve for anterior lip histogram value in predicting success of induction indicated a significant relationship with successful induction. The best cutoff value was 69.4. Conclusion: The value of anterior lip histogram associates significantly with the prediction of successful induction in nulliparous women. But, Bishop score and other sonographic measurement of cervical length, volume, and posterior lip histogram have no predictive values for successful induction.

Inhibition of HDACs and Aminopeptidases Is Highly Synergistic in Myeloma Cells Resulting in Cell Death Via the Upregulation of BIRC3, a Key Mediator of NF-KappaB Signalling.

Abstract 607Myeloma cells rely on a number of mechanisms to maintain their survival, including the silencing of genes that would normally check uncontrolled proliferation and lead to apoptosis. Epigenetic alterations such as histone deacetylation contribute significantly to the pathogenesis of both solid and haematological malignancies and are associated with the silencing of tumour suppressor genes. Pan-histone deacetylase inhibitors (HDACs), such as SAHA, panobinostat, depsipeptide, and numerous others have been published to have anti-myeloma activity. Here we report the effects of a novel pan HDAC inhibitor, CHR-3996, on multiple myeloma cells. Results: CHR-3996 was potently cytotoxic against a panel of myeloma cell lines (H929, KMS11, LP-1, MM1s, RPMI-8226, and U266) with a GI50 ranging from 9 to 65 nM and was equally as potent in primary patient myeloma cells. The loss of cell viability was associated with an increase in apoptotic cells; EndonucleaseG and Noxa were both up-regulated and caspase 9 was cleaved. Furthermore a decrease in apoptosis was demonstrated in the presence of a pan-caspase inhibitor indicating cell death is largely dependent on caspase-mediated apoptosis. There was no effect on bone marrow stromal (BMS) cell viability but there was an observed decrease in the IL-6 and VEGF the BMS cells secrete, both of which promote the growth and survival of myeloma cells in the bone marrow microenvironment. CHR-3996 caused an increase in acetylated H3K9 but there was minimal change in the levels of ubiquitinated proteins in the cell or acetylated alpha-tubulin, indicating low activity against HDAC6 or the proteasome (also demonstrated by an assay to specifically measure proteasome function). In addition to induction of apoptosis, cell cycle analysis showed an increased proportion of cells in G0/G1 indicating cell cycle arrest. In-keeping with data from treating myeloma cells with SAHA and panobinostat, an increase in cell cycle inhibitor p21 was observed. Gene expression profiling (GEP) identified changes in cell cycle regulators, indications of increased cell stress (elevated CHOP, ATF3, and TAO kinase 3), repression of Wnt (up-regulation of NLK, GSK3beta) and mTOR (decreased 4E-BP1) signalling, and changes in key pro- and anti-apoptotic proteins (for example SMAD3, BCL2, BIM, BID, and BIRC5). When used in combination studies CHR-3996 was highly synergistic in vitro with tosedostat, an aminopeptidase inhibitor, which we have previously shown to have anti-myeloma activity via the induction of the amino acid deprivation response and autophagy. One of the largest changes the GEP analysis identified was BIRC3 (CIAP2), an inhibitor of NF-kappaB signalling, which increased 23.5 fold with CHR-3996 as a single agent and over 100-fold when added to H929 cells in combination with tosedostat. Additionally both CHR-3996 and todestat independently up-regulated expression of members of the IkappaB family. Altered expression and nuclear localisation of canonical and non-canonical NF-kappaB family members were observed by immuno-fluorescence and immunoblotting, suggesting targeting of NF-kappaB signalling as a reason for the high degree of synergy between these two compounds. Early data suggest oral CHR-3996 is effective in a subcutaneous in vivo myeloma model and, reflecting the in vitro data, there is a degree of synergy when administered with tosedostat. Conclusions: The novel compound CHR-3996 is a potent HDAC inhibitor that leads to increased H3K9 acetylation but has no detectable activity against HDAC6 or proteasome activity in myeloma. It induces apoptosis of myeloma cells without affecting BMS cell viability. CHR-3996 shows a very high degree of synergy with an aminopeptidase inhibitor, tosedostat, potentially through targeting the NF-kappaB pathway. It has exciting therapeutic potential either as a mono-therapy or in combination with other agents. Disclosures:Krige:Chroma Therapeutics Ltd: Employment, Equity Ownership. Hooftman:Chroma Therapeutic Ltd: Employment, Equity Ownership. Drummond:Chroma therapuetics: Employment, Equity Ownership.

B9 A NOVEL, AUTOMATED METHOD OF MEASURING THE MOUSE BEHAVIOURAL SATIETY SEQUENCE

1CNS group, Paradigm Therapeutics Ltd, Cambridge, UK 2The Vernalis Group of Companies, Winnersh, Berkshire, UK E-mail: [email protected] Abstracts of the 11th Biennial EBPS Meeting

Evaluation of a novel heterologous PrimeBoost immunotherapy in stage III/IV metastatic melanoma patients

2570 Background: This study evaluates the safety, immunogenicity and clinical response (by RECIST criteria) of increasing doses of DNA plasmid (DNA.Mel3) and MVA viral vector (MVA.Mel3) containing 7 human CTL epitopes from 5 melanoma antigens (Tyrosinase, melan-A, MAGE-1, MAGE-3, NY-ESO-1). Methods: 31 HLA-A2 positive patients with measurable stage III/IV melanoma were enrolled. Immunisations were 3 weeks apart. Epitope-specific CD8+ T cell responses were evaluated using ex vivo tetramer staining and interferon gamma (IFN-γ) ELISPOT assays. Results: At week 16, 20 currently completed patients showed no dose limiting toxicity with only mild to moderate injection site reactions predominantly reported post MVA.Mel3. In the low dose groups (groups 1–3), 8/15 immunologically evaluable patients showed a significant increase of melanA CD8+ tetramer+ T cells. Of these 8, 3 also showed a significant increase of IFN-γ secreting T cells against melanA, tyrosinase leader, and/or NY-ESO-1. One clinical responder, confirmed at week 28, had ELISPOT responses against both melanA and tyrosinase leader. Additionally, of two patients with stable disease at week 16 one showed increases in melanA CD8+ tetramer+ T cells. Conclusions: Low dose heterologous PrimeBoost immunotherapy was safe and stimulated immune responses in >50% of metastatic melanoma patients treated. Immune responses against multiple antigens were observed in the partial clinical responder. Complete 16 week data from the high dose groups (10e8–10e9 pfu) will be available for presentation. Author Disclosure Employment or Leadership Consultant or Advisory Role Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration Oxxon Therapeutics Ltd. Oxxon Therapeutics Ltd.

A Widely Available Method for the Assessment of Aerosol Delivery in Cystic Fibrosis

Whilst nebulisers are commonly used in the treatment of cystic fibrosis (CF), nebulised aerosol lung deposition in individual patients is not routinely assessed in clinical practice. The present study was designed to evaluate whether a comparative measurement of aerosol lung deposition from nebulisers using a widely available scintigraphic method could be employed to assist the selection of the best system for individual patients. Lung deposition of the radiolabelled aerosol from the Pari LC Plus (Pari Medical Ltd) nebuliser and the HaloLite Adaptive Aerosol Delivery (AAD®) system (Profile Therapeutics Ltd) was measured using planar scintigraphy in 10 healthy volunteers and 6 CF patients. The HaloLite AAD® delivered on average 2.1 times ( P=0.003) as much aerosol to the lungs compared with Pari LC Plus. Only two subjects had higher lung deposition from Pari LC Plus than HaloLite AAD® system. There was marked inter-individual variation in the deposition pattern in CF patients. The aerosol deposition from HaloLite AAD® had higher central distribution than that obtained with the Pari LC Plus. The overall intersubject variability of the delivered dose was 56% with Pari LC Plus and 24% with HaloLite AAD® ( P<0.05). The measurement of aerosol deposition from nebulisers can be performed using a simple and widely available methodology, and may improve nebuliser selection in CF patients.

5476995 Peptide production : Clark Anthony; Lathe Richard Edinburgh, United Kingdom assigned to PPL Therapeutics (Scotland) Ltd

5476995 Peptide production : Clark Anthony; Lathe Richard Edinburgh, United Kingdom assigned to PPL Therapeutics (Scotland) Ltd