• Synergistic effects of imatinib (STI 571) in combination with chemotherapeutic drugs in head and neck cancer.

      Bruce, Iain A; Slevin, Nicholas J; Homer, Jarrod J; McGown, Alan T; Ward, Timothy H; Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK. (2005-08)
      The tyrosine kinase inhibitor imatinib (STI 571; glivec) is a potent inhibitor of bcr-abl, c-kit and platelet-derived growth factor receptors. Imatinib was evaluated both alone and in combination with established chemotherapeutic agents in adenoid cystic carcinoma (ACC) primary cultures and established cell lines representing squamous cell carcinoma of the head and neck (HNSCC). Over 90% of ACC tumors are c-kit-positive, and these primary cultures proved to be of short-term usefulness in assessing chemosensitivity. Interaction was determined over a wide range of drug combinations using a statistical three-dimensional analysis model. Both ACC short-term cultures and HNSCC cell lines were demonstrated to have a response ranging from additive to synergistic when imatinib and cisplatin were combined. The interaction of imatinib on cisplatin-induced DNA cross-linking was further investigated using the comet-X assay. In contrast, significant antagonism was observed when imatinib and gemcitabine were combined. Since gemcitabine is activated by deoxycytidine kinase (dCK), the effect of imatinib on this enzyme was investigated. A dose-dependent inhibition of dCK was observed, highlighting this kinase as a possible additional secondary molecular target for imatinib. This work demonstrates a synergistic interaction between cisplatin and imatinib, which may prove to be clinically relevant in the future management of both ACC and HNSCC.
    • Synergistic interactions in haemopoiesis: biological implications and clinical use.

      Dexter, T Michael; Christie Hospital, Withington, Manchester, UK. (1993)
      Growth factors promote the survival and proliferation of haemopoietic stem and progenitor cells, and in their absence the haemopoietic cells undergo apoptosis and die. The results of studies reported here indicate that multipotent stem cells have receptors for most, if not all, of the growth factors, but that even saturated binding of the receptors for a single growth factor is not sufficient to transduce an effective stimulus for the proliferation of these cells (possibly due to very low receptor numbers). However, when the growth factors are combined synergistic effects can be seen. Studies in which stem cell factor was used in combination with other growth factors showed that stem cell factor allowed the survival of stem cells, while a second growth factor (granulocyte-macrophage colony-stimulating factor) stimulated the stem cells to develop normally. Stem cell factor was also shown to alter the dose-response relationships of developing haemopoietic cells for other growth factors.
    • A syngeneic mouse B-cell lymphoma model for pre-clinical evaluation of CD19 CAR T cells.

      Kueberuwa, Gray; Zheng, W; Kalaitsidou, Milena; Gilham, David E; Hawkins, Robert E; Manchester Cancer Research Centre Building, Department Cancer Sciences, University of Manchester (2018)
      The astonishing clinical success of CD19 chimeric antigen receptor (CAR) T-cell therapy has led to the approval of two second generation chimeric antigen receptors (CARs) for acute lymphoblastic leukemia (ALL) andnon-Hodgkin lymphoma (NHL). The focus of the field is now on emulating these successes in other hematological malignancies where less impressive complete response rates are observed. Further engineering of CAR T cells or co-administration of other treatment modalities may successfully overcome obstacles to successful therapy in other cancer settings. We therefore present a model in which others can conduct pre-clinical testing of CD19 CAR T cells. Results in this well tested B-cell lymphoma model are likely to be informative CAR T-cell therapy in general. This protocol allows the reproducible production of mouse CAR T cells through calcium phosphate transfection of Plat-E producer cells with MP71 retroviral constructs and pCL-Eco packaging plasmid followed by collection of secreted retroviral particles and transduction using recombinant human fibronectin fragment and centrifugation. Validation of retroviral transduction, and confirmation of the ability of CAR T cells to kill target lymphoma cells ex vivo, through the use of flow cytometry, luminometry and enzyme-linked immunosorbent assay (ELISA), is also described. Protocols for testing CAR T cells in vivo in lymphoreplete and lymphodepleted syngeneic mice, bearing established, systemic lymphoma are described. Anti-cancer activity is monitored by in vivo bioluminescence and disease progression. We show typical results of eradication of established B-cell lymphoma when utilizing 1st or 2nd generation CARs in combination with lymphodepleting pre-conditioning and a minority of mice achieving long term remissions when utilizing CAR T cells expressing IL-12 in lymphoreplete mice. These protocols can be used to evaluate CD19 CAR T cells with different additional modification, combinations of CAR T cells and other therapeutic agents or adapted for the use of CAR T cells against different target antigens.
    • The syntheses and properties of tricyclic pyrrolo[2,3-d]pyrimidine analogues of S6-methylthioguanine and O6-methylguanine

      Hornillo-Araujo, Ana R; Burrell, Adam J M; Aiertza, Miren K; Shibata, Takayuki; Hammond, David M; Edmont, Dolores; Adams, Harry; Margison, Geoffrey P; Williams, David M; Centre for Chemical Biology, Richard Roberts Building, Department of Chemistry, University of Sheffield, Brook Hill, Sheffield, UK S3 7HF (2006)
    • Synthesis and anticancer activities of 4-oxobenzopyrano[2,3-d]pyrimidines.

      Hadfield, John A; Pavlidis, V H; Perry, P J; McGown, Alan T; Cancer Research Campaign Section of Drug Development and Imaging, Paterson Institute for Cancer Research, Manchester, UK. jhadfield@picr.man.ac.uk (1999-07)
      Several 2-aryl-4-oxoxbenzopyrano[2,3-d]pyrimidines have previously been shown to exhibit in vivo antitumor activity in mice with P388 lymphocytic leukemia. In the present study, a series of novel substituted benzopyrano[2,3-d]pyrimidines have been prepared and tested for cytotoxic activity against a panel of cancer cell lines including the P388 lymphocytic leukemia cell line. The unsubstituted parent compound, some methoxylated derivatives and a cyclohexyl derivative all exhibited potent cytotoxic activity (IC50 values 0.3-0.64 microM). A number of derivatives, including the unsubstituted parent pyrimidine, were shown to cause a significant perturbation in cell cycle kinetics with an observed 2- to 3-fold increase in cells in the G2/M phase of the cell cycle. Furthermore, a polymethoxylated derivative, 2-(3,4,5-trimethoxyphenyl)-9-methoxy-4-oxo-2,3-dihydrobenzopyrano[ 2,3-d]pyrimidine 13, was shown to be selectively active against a number of human ovarian cell lines.
    • Synthesis and anticancer activity of fluorinated analogues of combrestatin A-4.

      Lawrence, Nicholas J; Hepworth, Lucy A; Rennison, David; McGown, Alan T; Hadfield, John A (2003)
    • Synthesis and antiproliferative activity of unsaturated quinoline derivatives.

      Montgomery, Gerard J; McKeown, Paul; McGown, Alan T; Robins, David J; Department of Chemistry, University of Glasgow, UK. (2000-06)
      In our previous work Knoevenagel condensation of quinoline 2-, 3- and 4-carbaldehyde with malononitrile derivatives was used to produce a series of heteroarylidene malononitrile derivatives. Some of these heteroaromatic tyrphostins were potent inhibitors of the epidermal growth factor (EGF) receptor kinase. This work has now been extended by using 6-, 7-, and 8-quinolinecarbaldehyde to prepare 23 new quinoline-tyrphostins 1-23. Most of these compounds were moderately active against the MCF7 breast cancer cell line. The order of potency was 7- > 6 > 8-substituted quinoline, which indicates that increased activity of the 7-substituted quinolines is associated with electron deficiency at the 7-position in the quinoline ring. The most active compound, 12, formed from 7-quinolinecarbaldehyde and ethyl cyanoacetate, had an IC50 value of 2.3 microM. Compounds 1-23 showed similar IC50 values against the MCF7 and MCF7/ADR cell lines (the latter shows fourfold increased protein tyrosine kinase activity) except for the compounds 1 and 15 formed from 6-quinolinecarbaldehyde and malononitrile and 7-quinolinecarbaldehyde and cyanoacetamide, which showed a significant (11- and 42-fold, respectively) increase in potency against the MCF7/ADR cell line. Furthermore, no association was found between growth inhibition and inhibition of the EGFR protein tyrosine kinase (PTK), using a cell-free assay. In addition, new compounds were prepared from 2- and 4-quinolinecarbaldehyde with extended conjugation in the side chains (24-27) or with methoxypolyethoxyethyl esters in the side chain to increase water solubility (28 and 29). These compounds showed substantial cytotoxicity, with IC50 values in the range 1-25 microM, but similar values were observed against both cell lines. No association was found between inhibition of PTK and growth inhibition, again indicating that their mode of action may not be specific for the EGF receptor.
    • Synthesis and biological activity of analogues of ptilomycalin A

      Black, Gregory P; Coles, Simon J; Hizi, Amnon; Howard-Jones, Andrew G; Hursthouse, Michael B; McGown, Alan T; Loya, Shoshana; Moore, C G; Murphy, Patrick J; Smith, Nigel K; et al. (2001-05)
    • Synthesis and cell growth inhibitory properties of substituted (E)-1-phenylbut-1-en-3-ones

      Ducki, Sylvie W; Hadfield, John A; Hepworth, Lucy A; Lawrence, Nicholas J; Ching-Ying, Liu; McGown, Alan T (1997)
    • The synthesis and characterization of compounds of the type Hg[1-C(6)H(4)-2-C(H)=NC(6)H(5-n)R(n)](2).

      Flower, Kevin R; Howard, Victoria J; Naguthney, Shakil; Pritchard, Robin G; Warren, John E; McGown, Alan T; CRC Drug Development Group, Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK. k.r.flower@umist.ac.uk (2002-04-08)
      The organomercurial compounds Hg[1-C(6)H(4)-2-C(H)=NC(6)H(5-n)R(n)](2) (R = 4-NMe(2), 6a; 4-Me, 6b; 4-I, 6c; 4-NO(2), 6d; 2-(i)Pr, 6e; 2-Me, 6f; 2,6-(i)Pr(2), 6g; 2,6-Me(2), 6h) have been prepared in good overall yield from 2-bromobenzaldehyde. All of the compounds have been characterized by elemental analysis, (1)H NMR, (13)C[(1)H] NMR, and infrared spectroscopy. In addition, compounds 6a [C(30)H(30)HgN(4), triclinic, P, a = 6.20000(10) A, b = 9.2315(2) A, c = 10.9069(3) A, alpha = 85.8510(10) degrees, beta = 89.3570(10) degrees, gamma = 87.206(2) degrees, Z = 1], 6b [C(28)H(24)HgN(2), monoclinic, P2(1)/c, a = 12.8260(5) A, b = 14.0675(4) A, c = 6.1032(2) A, beta = 90.0990(10) degrees, Z = 2], 6g [C(38)H(44)HgN(2), triclinic, P, a = 8.2626(2) A, b = 9.8317(2) A, c = 11.8873(3) A, alpha = 103.6650(10) degrees, beta = 109.3350(10) degrees, gamma = 104.627(2) degrees, Z = 1], and 6h [C(30)H(28)HgN(2), monoclinic, P2(1)/c, a = 12.5307(2) A, b = 10.9852(2) A, c = 18.2112(2) A, beta = 104.0190(10) degrees, gamma = 87.206(2) degrees, Z = 4] have been characterized by low-temperature single-crystal X-ray diffraction studies, and two different molecular geometries about the central mercury atom have been observed; intramolecular contacts suggest a van der Waals radius for Hg of 2.1-2.2 A.
    • Synthesis and evaluation of double bond substituted combretastatins.

      Hadfield, John A; Gaukroger, Keira; Hirst, Nicholas; Weston, Anna P; Lawrence, Nicholas J; McGown, Alan T; Centre for Molecular Drug Design, Cockcroft Building, University of Salford, Manchester M5 4WT, UK. j.a.hadfield@salford.ac.uk (2005-06)
      A series of combretastatins substituted with epoxides, amides and small alkyl groups has been synthesised and evaluated for cytotoxicity and their ability to inhibit the assembly of tubulin. The methyl and ethyl substituted phenols 36, 44 have shown potent antimitotic effects whilst exhibiting reduced cytotoxicity.
    • The synthesis and properties of tricyclic analogues of S6-methylthioguanine and O6-methylguanine.

      Hornillo-Araujo, Ana R; Burrell, Adam J M; Aiertza, Miren K; Shibata, Takayuki; Hammond, David M; Edmont, Dolores; Adams, Harry; Margison, Geoffrey P; Williams, David M; Centre for Chemical Biology, Department of Chemistry, University of Sheffield, Sheffield, United Kingdom. (2007)
      The syntheses of novel tricyclic pyrrolo[2,3-d]pyrimidine analogues of O(6)-methylguanine and S(6)-methylthioguanine are described. The crystal structures and pK(a) values of these analogues are reported. In a standard substrate assay with the human repair protein O(6)-methylguanine-DNA methyltransferase (MGMT) only the oxygen-containing analogue displayed activity.
    • The Synthesis of (E) and (Z)-Combretastatins A-4 and a Phenanthrene from Combretum caffrum

      Lawrence, Nicholas J; Ghani, Fazni A; Hepworth, Lucy A; Hadfield, John A; McGown, Alan T; Pritchard, Robin G; Department of Chemistry, UMIST, PO Box 88, Manchester, M60 1QD, UK (1999)
    • Synthesis of [18F]fluoroacetaldehyde. Application to [18F]fluoroethylation of benzylamine under reductive alkylation conditions

      Prenant, C; Gillies, James M; Bailey, J; Chimon, G N; Smith, N; Jayson, Gordon C; Department of Medical Oncology, Christie Hospital NHS Trust, Wilmslow Road, Manchester M20 4BX, UK (2008)
    • A synthesis of crambescidin 359.

      Moore, C G; Murphy, Peter J; Williams, Harri L; McGown, Alan T; Smith, Nigel K (2003)
    • Synthesis of cytotoxic furonaphthoquinones: Regiospecific synthesis of diodantunezone and 2-ethylfuronaphthoquinones.

      Perry, P J; Pavlidis, V H; Hadfield, John A; Department of Chemistry, De Montfort University, The Gateway, Leicester LE1 9BH. UK. (1997)
    • Synthesis of glycosaminoglycans by human skin fibroblasts cultured on collagen gels.

      Gallagher, John T; Gasiunas, Nijole; Schor, Seth L; Cancer Research Campaign Department of Medical Oncology, Christie Hospital and Holt Radium Institute Manchester M20 9BX, U.K. (1980-08-15)
      A comparison has been made of the synthesis of glycosaminoglycans by human skin fibroblasts cultured on plastic or collagen gel substrata. Confluent cultures were incubated with [3H]glucosamine and Na235SO4 for 48h. Radiolabelled glycosaminoglycans were then analysed in the spent media and trypsin extracts from cells on plastic and in the medium, trypsin and collagenase extracts from cells on collagen gels. All enzyme extracts and spent media contained hyaluronic acid, heparan sulphate and dermatan sulphate. Hyaluronic acid was the main 3H-labelled component in media and enzyme extracts from cells on both substrata, although it was distributed mainly to the media fractions. Heparan sulphate was the major [35S]sulphated glycosaminoglycan in trypsin extracts of cells on plastic, and dermatan sulphate was the minor component. In contrast, dermatan sulphate was the principal [35S]sulphated glycosaminoglycan in trypsin and collagenase extracts of cells on collagen gels. The culture substratum also influenced the amounts of [35S]sulphated glycosaminoglycans in media and enzyme extracts. With cells on plastic, the medium contained most of the heparan sulphate (75%) and dermatan sulphate (> 90%), whereas the collagenase extract was the main source of heparan sulphate (60%) and dermatan sulphate (80%) from cells on collagen gels; when cells were grown on collagen, the medium contained only 5-20% of the total [35S]sulphated glycosaminoglycans. Depletion of the medium pool was probably caused by binding of [35S]sulphated glycosaminoglycans to the network of native collagen fibres that formed the insoluble fraction of the collagen gel. Furthermore, cells on collagen showed a 3-fold increase in dermatan sulphate synthesis, which could be due to a positive-feedback mechanism activated by the accumulation of dermatan sulphate in the microenvironment of the cultured cells. For comparative structural analyses of glycosaminoglycans synthesized on different substrata labelling experiments were carried out by incubating cells on plastic with [3H]glucosamine, and cells on collagen gels with [14C]glucosamine. Co-chromatography on DEAE-cellulose of mixed media and enzyme extracts showed that heparan sulphate from cells on collagen gels eluted at a lower salt concentration than did heparan sulphate from cells on plastic, whereas with dermatan sulphate the opposite result was obtained, with dermatan sulphate from cells on collagen eluting at a higher salt concentration than dermatan sulphate from cells on plastic. These differences did not correspond to changes in the molecular size of the glycosaminoglycan chains, but they may be caused by alterations in polymer sulphation.
    • The synthesis of proteoglycans by human T lymphocytes.

      Steward, William P; Christmas, Stephen E; Lyon, Malcolm; Gallagher, John T; CRC Dept. Medical Oncology, Christie Hospital, Manchester, U.K. (1990-05-22)
      We have examined the proteoglycans produced by highly-purified cultures of human T-lymphocytes. The proteoglycans were metabolically labelled with [35S]sulphate and analysed in cellular and medium fractions using DEAE-cellulose chromatography, gel filtration and specific enzymatic and chemical degradations. The results showed that the T cells synthesized a relatively homogeneous, proteinase-resistant chondroitin 4-sulphate proteoglycan that accumulated in the culture medium during a 48 h incubation period. The cellular fraction contained a significant amount of free chondroitin sulphate chains that were not secreted into the medium. These polysaccharides were formed by intracellular degradation of proteoglycan in a chloroquine-sensitive process, indicating a requirement for an acidic environment. In contrast to chondroitin sulphate derived from proteoglycan, chondroitin sulphates synthesized on the exogenous primer, beta-D-xyloside, were mainly secreted by the cells. beta-D-Xylosides caused an 8-fold stimulation in the synthesis of chondroitin sulphate, but decreased the synthesis of proteoglycan by about 50%. These proteoglycans contained shorter chondroitin sulphate chains than their normal counterparts. The results indicate that although proteoglycans are mainly secretory components in human T-cell cultures, a specific metabolic step leads to the intracellular accumulation of free glycosaminoglycans. Separate functions are likely to be associated with the intracellular and secretory pools of chondroitin sulphate.
    • Synthesis of water-soluble sugar derivatives of combretastatin A-4

      Brown, R; Fox, Brian W; Hadfield, John A; McGown, Alan T; Mayalarp, Stephen P; Pettit, G; Woods, J; Department of Chemistry, University of Manchester, Manchester M13 9PL (1995)