Fourth World Antibody-Drug Conjugate Summit

The 4th World Antibody Drug Conjugate (WADC) Summit, organized by Hanson Wade was held on February 29?March 1, 2012 in Frankfurt, Germany, which was also the location for the Antibody Drug Conjugate Summit Europe held in February 2011. During the one year between these meetings, antibody drug conjugates (ADCs) have confirmed their technological maturity and their clinical efficacy in oncology. Brentuximab vedotin (ADCETRIS^TM) gained approval by the US Food and Drug Administration in August 2011 and trastuzumab emtansine (T-DM1) confirmed impressive clinical efficacy responses in a large cohort of breast cancer patients. During the 4th WADC meeting, antibody-maytansinoid conjugates were showcased by representatives of ImmunoGen (T-DM1, SAR3419, lorvotuzumab mertansine/IMGN801, IMGN529 and IMG853) and Biotest (BT-062). Data on antibody-auristatin conjugates were presented by scientists and clinicians from Seattle Genetics and Takeda (brentuximab vedotin), Pfizer (5T4-MMAF), Agensys/Astella (AGS-16M8F), Progenics (PSMA-ADC) and Genmab (anti-TF ADCs). Alternative payloads such as calicheamicins and duocarmycin used for preparation of ADCs were discussed by Pfizer and Synthon representatives, respectively. In addition, emerging technologies, including site-directed conjugation (Ambrx), a protein toxin as payload (Viventia), hapten-binding bispecific antibodies (Roche), and use of light activated drugs (Photobiotics), were also presented. Last but not least, progresses in solving Chemistry Manufacturing and Control, and pharmacokinetic issues were addressed by scientists from Genentech, Pfizer, Novartis and Pierre Fabre.     

A Mechanistic Tumor Penetration Model to Guide Antibody Drug Conjugate Design

Antibody drug conjugates (ADCs) represent novel anti-cancer modalities engineered to specifically target and kill tumor cells expressing corresponding antigens. Due to their large size and their complex kinetics, these therapeutic agents often face heterogeneous distributions in tumors, leading to large untargeted regions that escape therapy. We present a modeling framework which includes the systemic distribution, vascular permeability, interstitial transport, as well as binding and payload release kinetics of ADC-therapeutic agents in mouse xenografts. We focused, in particular, on receptor dynamics such as endocytic trafficking mechanisms within cancer cells, to simulate their impact on tumor mass shrinkage upon ADC administration. Our model identified undesirable tumor properties that can impair ADC tissue homogeneity, further compromising ADC success, and explored ADC design optimization scenarios to counteract upon such unfavorable intrinsic tumor tissue attributes. We further demonstrated the profound impact of cytotoxic payload release mechanisms and the role of bystander killing effects on tumor shrinkage. This model platform affords a customizable simulation environment which can aid with experimental data interpretation and the design of ADC therapeutic treatments.     

Preparation and Retrograde Axonal Transport of an Antiviral Drug/Horseradish Peroxidase Conjugate

Recurrent Herpes simplex virus infections of the cornea are currently treated with antiviral drugs administered locally during periods of active infection. We have examined the feasibility of an alternate approach to treating recurrent infection, that of delivering an antiviral drug by axonal transport to cell somata in the trigeminal ganglion where the latent virus is thought to reside. We have coupled an antiviral drug to horseradish peroxidase (HRP), a protein which is readily transported retrogradely in corneal sensory axons to ganglion cell somata in the trigeminal ganglion. The antiviral drug 5-iodo-5'-amino-2',5'-dideoxyuridine (AIDU) was synthesized with 125I and coupled to horseradish peroxidase by Schiff base formation between the 5' amino group and aldehydes generated on the carbohydrate of the HRP following short periodate oxidation. The useful ratio of AIDU:HRP was 2-9; higher ratios resulted in an insoluble product. Sodium dodecylsulfate (SDS) acrylamide gel electrophoresis of the drug-protein conjugate revealed considerable aggregation and the isoelectric point of HRP was changed from 8.1 to 4.4-5.8 following the coupling procedure. Despite evidence that the protein conformation was considerably altered, the specific enzymatic activity of the final product was 58% of native HRP and the drug-protein conjugate was still strongly transported retrogradely. Retrograde transport of this conjugate was demonstrated by autoraiography of the trigeminal ganglion neurons 24 h after corneal injection of the [125I]AIDU/HRP.     

Production,Binding Characteristics and Protective Immunity of Monoclonal Antibody to Pneumococcal Type-9V Conjugate

A monoclonal antibody (MAb) to pneumococcal type-9V polysaccharide (PS) was produced using PS conjugated to inactivated pneumolysin as the immunogen. The MAb to 9V PS was of the IgG₁ subclass. The antigen-antibody reaction increased rapidly at low concentrations and reached a plateau at 10 μg PS/ml as measured by nephelometry of the group 9 PS against 9V MAb binding. In contrast, the binding of group 9 PS against rabbit 9V antiserum (AS) increased proportionally and continued to increase up to the highest concentration of PS tested (20 μg PS/ml). The 9V MAb reacted with all group 9 PSs (9A, 9L, 9N and 9V) by immunodiffusion. In the homologous 9V Ag-MAb reaction, there were marked differences in the inhibition of binding by the cross-reactive 9L PS (19.2% inhibition) and the 9N PS (0.2%). In contrast, inhibition of the homologous 9V Ag-rabbit AS binding by cross-reactive 9L and 9N PSs ranged from 57.8 to 62.7%. Removal of the O-acetyl group from 9V PS by alkali hydrolysis resulted in decreased binding with rabbit 9V AS. However, the binding reaction with 9V MAb was less affected by the loss of O-acetyl content. The 9V MAb was both opsonic and passively protected young mice against challenge with type-9V pneumococci.     

Multivalency: Key Feature in Overcoming Drug Resistance with a Cleavable Cell-Penetrating Peptide-Doxorubicin Conjugate

Multivalency is often used in biological systems, to increase affinity and specificity through avidity. This inspired us to prepare a synthetic bioconjugate that mimics natural multivalent systems. It is composed of doxorubicin and two octaarginine cell-penetrating peptides, to strengthen the electrostatic interactions between the negatively charged glycosaminoglycans of the plasma membrane and the guanidinium groups of the arginine residues. The multivalent conjugate has improved cellular uptake and cytotoxicity, compared to a peptide-drug conjugate with only one polyarginine and as a result it can overcome drug resistance in Kelly-ADR cells. The synthetic approach and the multivalent structure reported here can be used further as model systems, to gain insight into the biological interaction of cell-penetrating peptides with artificial membranes or for the preparation of more complex multimers.     

A Mammalian Lost World in Southwest Europe during the Late Pliocene

Background

Over the last decades, there has been an increasing interest on the chronology, distribution and mammal taxonomy (including hominins) related with the faunal turnovers that took place around the Pliocene-Pleistocene transition [ca. 1.8 mega-annum (Ma)] in Europe. However, these turnovers are not fully understood due to: the precarious nature of the period''s fossil record; the “non-coexistence” in this record of many of the species involved; and the enormous geographical area encompassed. This palaeontological information gap can now be in part bridged with data from the Fonelas P-1 site (Granada, Spain), whose faunal composition and late Upper Pliocene date shed light on some of the problems concerning the timing and geography of the dispersals.

Methodology/Principal Findings

This rich fossil site yielded 32 species of mammals, among which autochthonous species of the European Upper Villafranchian coexist with canids (Canis), ovibovines (Praeovibos) and giraffids (Mitilanotherium) from Asia. Typical African species, such as the brown hyena (Hyaena brunnea) and the bush pig (Potamochoerus) are also present.

Conclusions/Significance

This assemblage is taxonomically and palaeobiogeographically unique, and suggests that fewer dispersal events than was previously thought (possibly only one close to 2.0 Ma) are responsible for the changes seen around 1.9–1.7 Ma ago in the fauna of the two continents.     

Boechera Summit 2011

Ecological model systems provide a conduit to understand the ecological impact of information gained from laboratory model species. Here, I review a 2011 meeting which focused on the systematic, ecological, evolutionary and developmental biology of the ecological model genus Boechera.     

Global Alzheimer Research Summit

We report here on the proceedings of the Global Alzheimer Summit that took place September 22–23, 2011 in Madrid, Spain. As Alzheimer disease (AD) is the leading cause of neurodegeneration in elderly individuals and as yet has no effective therapeutic option, it continues to stimulate global research interests. At the conference, leaders in the field of AD research provided insights into current developments in various areas of research, namely molecular mechanisms, genetics, novel aspects of AD research and translational research. Emphasis was also placed on the importance of biomarkers in the diagnosis of AD and development of current therapeutic strategies.     

Antibody Responses in Humans Infected with Newly Emerging Strains of West Nile Virus in Europe

Infection with West Nile Virus (WNV) affects an increasing number of countries worldwide. Although most human infections result in no or mild flu-like symptoms, the elderly and those with a weakened immune system are at higher risk for developing severe neurological disease. Since its introduction into North America in 1999, WNV has spread across the continental United States and caused annual outbreaks with a total of 36,000 documented clinical cases and ∼1,500 deaths. In recent years, outbreaks of neuroinvasive disease also have been reported in Europe. The WNV strains isolated during these outbreaks differ from those in North America, as sequencing has revealed that distinct phylogenetic lineages of WNV concurrently circulate in Europe, which has potential implications for the development of vaccines, therapeutics, and diagnostic tests. Here, we studied the human antibody response to European WNV strains responsible for outbreaks in Italy and Greece in 2010, caused by lineage 1 and 2 strains, respectively. The WNV structural proteins were expressed as a series of overlapping fragments fused to a carrier-protein, and binding of IgG in sera from infected persons was analyzed. The results demonstrate that, although the humoral immune response to WNV in humans is heterogeneous, several dominant peptides are recognized.     

吸毒人群中HIV/HCV核酸和抗体关系的分析

研究了高危人群中HIV/HCV核酸和抗体的关系。从新疆地区采集吸毒人群的血样,并对其进行HIV/ HCV核酸和抗体的检测。320例吸毒人员血浆样品中HCV抗体阳性为80．3％,HIV抗体阳性率为41．9％,HIV 和HCV共感染者为38．3％。HIV RNA与抗体的总符合率为98．8%,在186例HIV抗体阴性样品中可能有2例 为HIV感染的窗口期。HCV抗体和HCV RNA的阳性符合率为92．6％,HCV RNA与HCV抗体的总符合率为 90．0％,以上结果说明在HIV／HCV的高流行区进行HIV／HCV核酸检测可以发现病毒感染的窗口期,而约8% 的HCV抗体阳性样品为病毒核酸阴性,也值得进一步研究。     

葡萄糖-阿司匹林偶联物的合成

目的:为了改善阿司匹林药效,降低其毒副作用,探索出一条操作简便、成本低廉的糖基化方法,研制出一种副作用低的缓释药物葡萄糖-阿司匹林偶联物。方法:本文采用吡啶、浓NaOH溶液和浓硫酸作催化剂,在常压、不同温度、不同n(乙酰水杨酰氯):n(葡萄糖)条件下,合成葡萄糖-阿司匹林共价偶联物,通过FT-IR、~1HNMR对其结构进行了表征。研究催化剂、反应温度以及反应物料比对酯化反应的影响,同时利用紫外分光光度法测定偶联物的接枝率。结果:在常压,50℃,n(乙酰水杨酰氯):n(葡萄糖)=5:1,吡啶为催化剂条件下,偶联物的接枝率可达到54.9%。结论:在此反应中,吡啶是一种良好的催化剂;50℃是该反应的最佳反应温度;n(乙酰水杨酰氯):n(葡萄糖)=5:1时,偶联物的接枝率和乙酰水杨酰氯的转化率都比较高。     

Cellular HIV-1 DNA Levels in Drug Sensitive Strains Are Equivalent to Those in Drug Resistant Strains in Newly-Diagnosed Patients in Europe

Background

HIV-1 genotypic drug resistance is an important threat to the success of antiretroviral therapy and transmitted resistance has reached 9% prevalence in Europe. Studies have demonstrated that HIV-1 DNA load in peripheral blood mononuclear cells (PBMC) have a predictive value for disease progression, independently of CD4 counts and plasma viral load.

Methodology/Principal Findings

Molecular-beacon-based real-time PCR was used to measure HIV-1 second template switch (STS) DNA in PBMC in newly-diagnosed HIV-1 patients across Europe. These patients were representative for the HIV-1 epidemic in the participating countries and were carrying either drug-resistant or sensitive viral strains. The assay design was improved from a previous version to specifically detect M-group HIV-1 and human CCR5 alleles. The findings resulted in a median of 3.32 log₁₀ HIV-1 copies/10⁶ PBMC and demonstrated for the first time no correlation between cellular HIV-1 DNA load and transmitted drug-resistance. A weak association between cellular HIV-1 DNA levels with plasma viral RNA load and CD4⁺ T-cell counts was also reconfirmed. Co-receptor tropism for 91% of samples, whether or not they conferred resistance, was CCR5. A comparison of pol sequences derived from RNA and DNA, resulted in a high similarity between the two.

Conclusions/Significance

An improved molecular-beacon-based real-time PCR assay is reported for the measurement of HIV-1 DNA in PBMC and has investigated the association between cellular HIV-1 DNA levels and transmitted resistance to antiretroviral therapy in newly-diagnosed patients from across Europe. The findings show no correlation between these two parameters, suggesting that transmitted resistance does not impact disease progression in HIV-1 infected individuals. The CCR5 co-receptor tropism predominance implies that both resistant and non-resistant strains behave similarly in early infection. Furthermore, a correlation found between RNA- and DNA-derived sequences in the pol region suggests that genotypic drug-resistance testing could be carried out on either template.